Examination of Firearms Review: 2007 to 2010 - Interpol

16th International Forensic 

Science Symposium 

Interpol – Lyon 

5 th - 8 th October 2010 

Review Papers 

Edited by Dr. Niamh Nic Daéid 

Centre for Forensic Science, University of Strathclyde, 

Glasgow, UK 

  I  

 

Table of contents 

PREFACE 

Mrs. Dominique Saint-Dizier,Ministere de l’Interieur, France. 

Chairman of the Organising Committee 

CRIMINALISTICS 1 

Technical co-ordinator :- Prof. Kimmo Himberg, 

National Institute of Forensic Science, Finland. 

Examination of Firearms 2-47 

Examination of Marks 48-86 

Examination of Glass 87-112 

Examination of Paint 113-181 

Examination of Fibres and textiles 182-200 

Forensic Geology 201-220 

HUMAN IDENTIFICATION 221 

Technical co-ordinator :- Mr Kevin O’ Sullivan, 

Forensic Science Service, UK. 

Fingermarks and Other Impressions Left by the 222-319 

Human Body 

Biological evidence 320-345 

Examination of Questioned documents/Handwriting 346-377 

MEDIA EVIDENCE 378 

Technical co-ordinator :- Dr. Peter Pfefferli, 

Forensic Science Institute Zurich Switzerland. 

Audio Analysis 379-396 

Digital Analysis 397-418 

CHEMICAL EVIDENCE 419 

Technical co-ordinator :- Dr. Niamh Nic Daeid, 

Centre for Forensic Science, University of Strathclyde, Scotland, UK. 

Fire cause and fire debris analysis 420-452 

Analysis and Detection of Explosives and Explosives 

Residues 453-523 

Hazardous materials 524-556 

Environmental forensics 

DRUGS AND TOXICOLOGY 608 

Technical co-ordinator :- Mr. Nelson Santos, 

Drug Enforcement Agency, USA. 

Toxicology 609-734 

Drugs 735-834 

  II 

Preface 

Forensic Science is playing an increasingly key role in investigations at the 

National level as well as in International police cooperation. For years, 

INTERPOL has understood the importance of the contribution of forensic 

science and the tri-annual International Interpol Forensic Science Sympoium 

organizes its 16 th conference this year. 

Its purpose remains the same : to bring together senior managers from 

member states, and to provide a forum to facilitate : 

− the presentation of advances made in scientific methods over the 

previous three (3) years, and to provide a look into future forensic 

needs and advances; 

− the exchange of information which will enhance scientific methods in 

criminal investigation and the administration of justice; 

− the discussion of problem areas encountered by member states and 

the possible provision of solutions; and 

− the exchange and pooling of ideas for future progress. 

The symposium Proceedings on this CD concentrate on the Review Papers 

prepared by the Coordinating Laboratories, which highlight and summarise 

advances in the various evidence types. The various evidence areas are 

grouped into five (5) major areas: 

Criminalistics 

Firearms; 

Toolmarks; 

Paint and glass; 

Fibres; 

Forensic geology. 

Human Identification 

Fingerprints. 

Biological evidence; 

Document examination / Handwriting; 

Media Evidence 

Audio Visual; 

Digital evidence; 

Chemical evidence 

Fire investigation; 

Explosives; 

Hazardous materials; 

  III 

Drugs and Toxicology 

Drugs; and 

Toxicology; 

As within the previous conferences, together with these main contributions, 

three (3) thematic panels will be held during this 16 th meeting on the topics of 

‘Environmental Crime’, ‘Biometrics and Forensic Identification’, and ‘Forensic 

Science : Effectiveness and Efficiency’. 

IIFSS is only possible with the support of Interpol and the General Secretary, 

Ronald Noble. Interpol staff coordinated all aspects of Interpol’s involvement 

from the meeting announcements to registration, the meeting venue and 

publication of the proceedings of the meeting. 

To make IIFSS 2010 possible it took significant effort by the Organising 

Committee, each Coordinating Laboratory and the review paper authors. 

The Organising Committee especially thanks Mr Simon Dzidrovski, Mrs Kim 

Legg and Mr Antonio Farelo from Interpol who act as the principle contact 

person for the Organising Committee. 

This CD would also not be possible without the work of Dr Niamh Nic Daéid 

who edits the submitted papers and prepares the proceedings. 

Mrs Dominique Saint-Dizier 

Chair 

IFSS Organising Committee 

  IV 

CRIMINALISTICS 

  1 

Examination of Firearms 

Review: 2007 to 2010 

Sébastien Charles PhD, Jan De Ceuster PhD, 

Patrick De Smet PhD, Bart Nys PhD 

INCC-NICC 

Chaussée de Vilvorde 100 

B-1120 Brussels 

Belgium 

Author responsible for correspondence: Sébastien Charles 

2

TABLE OF CONTENTS  

1.  INTRODUCTION  5  

2.  FIREARMS – BALLISTICS  5  

2.1  Firearms Identification  5  

2.1.1  Validation studies and statistical foundations  5  

2.1.2  Parameters that affect the identification process  7  

2.1.3  Identifications based on unusual markings  9  

2.1.4  Proficiency testing  10  

2.1.5  Instrumental methods  11  

2.1.6  Court rulings  14  

2.2  Firearms & ammunition miscellaneous reports  16  

2.2.1  Firearms  16  

2.2.2  Ammunition  16  

2.2.3  Recovery of obliterated serial numbers  16  

2.3  Legislation and other efforts  17  

2.3.1  Microstamping  17  

2.3.2  Glock EBIS barrel  18  

2.3.3  Bore lapping  18  

2.4  Shooting Incident Reconstruction  19  

2.4.1  Research  19  

2.4.2  Case reports  20  

2.4.3  Trace analysis  20  

2.5  Wound Ballistics  21  

2.5.1  Statistics  21  

2.5.2  Case reports  23  

2.5.3  Research  25  

2.6  Training Material & Books  27  

3  FIREARMS – CHEMISTRY  27  

3.1  Inorganic GSR  27  

3.1.1  Lead‐free ammunition  27  

3.1.2  Trends and comparisons  28  

3.1.3  Non‐GSR sources of GSR‐like particles  28  

3.1.4  Interpretation of analysis results and the application of Bayesian principles  29  

3.1.5  Quality  31  

3.2  Estimation of shooting distance and bullet hole characterization  32  

3.2.1  General considerations  32  

3.2.2  Colour tests  32  

3.2.3  Development of analytical techniques  34  

3.3  Organic GSR  34  

3

3.3.1  Screening of gun propellant powder components  35  

3.3.2  Instrumental methods  35  

3.4  Book review  36  

4.      REFERENCES  37  

4

1. Introduction 

This review paper covers advances in scientific methods applied to Firearms 

reported since the 15 th Interpol Forensic Science Symposium in October 2007. A 

literature search was conducted covering articles published in the main forensic 

journals since mid-2007. Two domains are distinguished: Ballistics and 

Chemistry. 

2. Firearms – Ballistics 

2.1 Firearms Identification 

In their report of 2009, the National Academy of Science made 13 

recommendations in regard to “Strengthening Forensic Science in the United 

States: A Path Forward.” (1) While those recommendations are very general, 

some of them relate to the discipline of firearms and tool marks identification. The 

Association of Firearms and Tool Marks Examiners (AFTE) has taken a stand on 

the issues and recommendations raised by NAS (2). Also the National 

Association of Criminal Defence Lawyers (NACDL) was preparing a position in 

the previously mentioned NAS report. NACDL raises seven areas of concern to 

which AFTE was given the opportunity to comment on their draft positioning 

paper (3). 

2.1.1 Validation studies and statistical foundations 

It has always been a desire to strengthen the scientific foundations of firearms 

identification so as to increase its acceptance in court. 

Buckelton et al. have considered the experimental design for acquiring relevant 

data to address the issue of comparing consecutively manufactured tools and 

firearms (4). This is interesting as comparing consecutively manufactured tools 

and firearms constitute the closest situation that one could encounter in known 

non matches. The authors pinpoint some criticisms to this method: 1/ most 

studies of consecutively manufactured tools use pristine substrate and optimal 

conditions; 2/ the test is seldom blind. The authors make suggestions for test 

designs to get more realistic results. 

One such study about consecutively manufactured extractors was published by 

Lyons (5). The author collected ten extractors from Caspian Arms. These were 

consecutively assembled in a Colt 1911A1 pistol. After that the pistol was fired 

with each of the extractors in place. The cartridge cases were collected and intercompared. 

It was demonstrated that there were enough individual characteristics 

to distinguish between the consecutively manufactured extractors. However, it 

was also noted that sub-class characteristics carried over from extractor to 

extractor. A test was set up consisting of multiple sets with 2 knowns and 1 

5

questioned. The sets were then distributed to volunteering examiners and the 

results collected. The error rate got as low as 1.2%. 

Hamby and Thorpe have also organised an experiment involving the 

identification of 617 different Glock pistols (6). Part of the experiment was to 

determine if an automated IBIS system has the ability to evaluate test fired 

cartridge cases to each other and to the exclusion of other like-type cartridge 

cases. It was found that none of the 617 test samples were associated to one 

another. In this research project, examination of the fired cartridge cases against 

each other, validated that each was identifiable and unique. This research 

supports the concept of individualizing fired cartridge cases to the firearm that 

fired them and should be of value in supporting the criteria for identification for 

firearm identification. 

Uchiyama created a test involving one hundred rounds with several brands of 

ammunition (7). The reproducibility of the markings was assessed using a home 

made image retrieval and identification system. Also, the extent of agreement 

was critically evaluated and described using a quantitative CMS technique. The 

author fired different brands of ammunition, also frangible bullets. To start with, 

the author observed a great variation in the shape of the land impression for 

different brands of ammunition. Examiners may potentially eliminate bullets from 

different brands of ammunition in the early stage of comparison if he/she has no 

prior knowledge of this variability. Secondly a prominent marking on one of the 

land impressions was observed to shift from shot to shot. This was possibly 

explained due to differences in bullet velocity and / or diameter. Identification 

between bullets from different manufacturers was generally difficult. As for the 

cartridge cases: the size of the primer’s areas of contact with the breech face and 

firing pin was different among each brand of ammunition but were consistent 

within the same make of ammunition. The number of parallel striations on 

primers decreased with successive firing. 

A comparable study was done by Gouwe, Hamby and Norris (8). This study was 

conducted to determine if the toolmarks on fired cartridge cases would change 

significantly during the firing of 10000 rounds from a Glock .40 S&W calibre 

pistol. Examination revealed that all 10000 cases could be identified to each 

other. Besides evaluating the comparisons using traditional pattern matching, the 

authors have also applied the CMS technique. Both provided the same results, 

however it is the opinion of the authors that CMS has the potential to provide the 

examiner with additional resources in today’s legal climate. 

Another massive study involved the identification of bullets fired from 10 

consecutively rifled 9 mm Ruger pistol barrels (9). 240 test sets were made and 

sent out to different forensic laboratories. 507 qualified participants returned their 

answering sheet. After elimination of some unusable items, all of the remaining 

7597 unknown fired bullets were correctly identified to the provided known 

6

ullets. Although one would expect bullets fired from consecutively rifled barrels 

to display subclass characteristics, these were not an issue. 

Neel and Wells published a comprehensive statistical analysis of striated tool 

mark examinations (10). The purpose of the paper was to quantify the difference 

between Known Matches (KM) and Known Non-Matches (KNM). Over 4000 

striated toolmark comparisons were examined for their Total Matching Lines 

(TML), Percent Matching Lines (%ML) and the size and frequency of various 

CMS runs. It was interesting that the TML did not always equal the number of 

matching striae tabulated into CMS groupings. It was concluded that TML and 

%ML are no good criteria for identification. Another point of interest was 

determining the mathematical difference between the most conservative 2D and 

3D KM to the best 2D and 3D KNM. The next step was to calculate whether or 

not these differences are statistically significant. Through Z-test calculations the 

authors evaluated the null hypothesis at the 99% confidence level that there is no 

statistically significant difference between the probabilities of, for example, a 2-5x 

CMS run in a 2D KM and a 1-6x CMS run in a 2D KNM. The null hypothesis was 

rejected at all times in favour of the alternative hypothesis. Furthermore, a 

discussion is given of what the “practical impossibility” would mean from a 

statistical point of view. The research demonstrates that not only can KM and 

KNM be differentiated on a microscope, but they can be differentiated statistically 

by comparing the probabilities of CMS run sizes. 

An identification is based on the comparison of specific regions of similarity and 

is sometimes under built by the criterion of the presence of a specific number of 

consecutively matching striae as a way to reduce the level of subjectivity. Howitt 

et al. have published a technical note on the calculation of the theoretical 

significance of matched bullets (11). This publication assesses the statistical 

likelihood of a particular correspondence of striae. The calculations are based on 

the resolution of striae that could be discerned through a typical comparison 

microscope. Consequently the method takes into account apparent differences in 

width of stria to attribute more uniqueness to them. Through statistical 

calculations of the probabilities for finding random sequences of line pairs, the 

authors come at the total probabilities of matching striation patterns. There is a 

close match between these theoretical results and the empirical results from Al 

Biasotti from the late 1950’s. The calculations thus provide a firm theoretical 

basis for the concept of CMS (consecutive matching striae), that was introduced 

by the latter. 

2.1.2 Parameters that affect the identification process 

A recent publication by Justine Davis goes in depth on primer cup properties and 

how they affect identification (12). Issues related to ammunition include the 

cartridge case material, the amount and type of propellant used and the primer 

cup hardness. The author discusses in great detail the physical properties of 

metals and the Hardness factor. A research project was set up to determine how 

7

these properties affect breech face marks. Seven different types of 9 mm Luger 

cartridges and four different firearms were used with four different breech face 

manufacturer marks. Hardness of the primer was determined both before and 

after firing. It was concluded that nickel plating added to the hardness of the 

primer cups and hardness significantly increased after the firing process. Fired 

cartridge cases were then compared to a breech face cast. The results 

demonstrated notable and consistent differences in marks on the primer cup. 

Overall, both nickel plated and brass primer cups of similar hardness values were 

obtaining marks similarly. It was noted though that primer cups with hardness 

values greater than the others were acquiring marks significantly different. 

Hardest and softest primers do not always behave in opposite manners and 

other variables are likely playing a part in the way cartridge cases are marking. 

Rayer has examined the effects of molybdenum disulfide coating on the 

formation of striations on fired bullets. Due to the sheen of the bullets, inhibiting 

reflective light during microscopic examination, there was a decreased capacity 

to identify two consecutive bullets fired from the same barrel (13). 

A second and third part of an extensive study on the effects of fire damage on 

the ability to make identifications was published. In the second part (14) 

Marsanopoli, Gerber and Dandridge determined whether a firearm will retain 

sufficient individual microscopic markings to allow an identification to be made 

after it had been subjected to a fire. The emphasis of this part lies on the 

cartridge cases. It remains possible to identify ammunition components that have 

been fired from a gun prior to it being subjected to a fire to those from the same 

firearms post-fire. However, there are many factors that will affect the potential 

for identification, including heat intensity, changes to the surface of the firearm, 

rust formation, pitting of the breech and water damage. Different areas of the 

cartridge cases will have to be used to make successful identifications. In the 

third part of their study (15), the authors compare burned firearms and 

ammunition components to determine if they retain identifiable individual 

microscopic characteristics. A first scenario simulated the case were bullets were 

recovered from a burned victim. They could be identified to each other and to 

reference bullets from the firearm pre-burn. However, if the firearm was burned 

too, many additional microscopic striae will be present on the post-burn test fires 

making it impossible to identify them to the bullets recovered from the carcass. It 

was worse for bullets that were placed in the car seats or on the floor of the truck. 

Post-burn, they did not have sufficient agreement of individual characteristics to 

identify them as having been fired from the same firearm nor was it possible to 

identify them back to the original test fires. The last scenario involved cartridge 

cases where it was found that individual markings are not always retained to 

enable positive identifications. Fire debris can sometimes conceal markings. A 

short case report on a fire damaged shotgun shell was published by Sarıbey and 

Hannam (16). 

8

A classic example of subclass characteristics was reported on by Rivera (17). 

The author demonstrates clear subclass agreement in breech face markings of 

two different Smith & Wesson SW40VE Sigma Pistols. However, the pistols do 

produce distinctive shear patterns that could be used to differentiate between 

them. 

Uchiyama studied the variation of markings on primers of cartridge cases fired by 

different .25 Semi-Automatic pistols (18). The study was undertaken to examine 

if one would be able to identify the make and model of the firearm, based on 

firing pin indentation and firing aperture differences. Large variations were 

observed, even for the same make and model of firearm. It was found that it 

would be difficult to predict the probable make of firearm from the average pin 

impression diameter and eccentricity, and from the firing pin aperture markings. 

Sarıbay, Hannam and Tarımcı (19) investigated whether or not class and 

individual characteristics of a firearm change over time and during extensive 

firing. They looked particularly at five Turkish pistols. It was found that there were 

slight changes in some of the individual and class characteristics; however, this 

was not statistically significant and did not affect the ability of the firearms 

examiner to match the first fired case to the last fired case for each firearm 

tested. 

2.1.3 Identifications based on unusual markings 

Jay Stuart examined the usefulness of bullet cannelures to link a scene bullet to 

a suspect box of ammunition (20). Three boxes of 50 Winchester cartridges were 

purchased for comparison. Two boxes were from the same lot whereas the other 

is from a different lot. Comparisons were conducted to determine the number of 

different knurled cannelures present in each box and if there were different 

knurled cannelures, could they be found in different boxes. It was observed that 

each of the three boxes contained cannelures made by four different knurling 

tools. Bullets were compared intra-box and inter-box. In addressing the 

hypothesis “knurled cannelures contain sufficient individual characteristics 

necessary for a positive identification to a specific tool”, it is clear that the 

Winchester bullets do indeed possess the individual characteristics necessary for 

comparison. With regard to the hypothesis “Bullets can be linked to a specific box 

or lot of ammunition through comparison”, the key word is specific. While it is true 

that bullet cannelures are identifiable, there are between 7500 and 84000 other 

boxes from a single production year in the market that could contain other bullets 

with matching cannelures. 

Clow suggest a method for the production of test specimens for the comparison 

of bullet seating marks (21). Sometimes an examiner is asked if it is possible to 

identify a bullet back to a cartridge case. Indeed, as the bullet is seated, a 

striated mark can be produced on the bearing surface starting at the base and 

running to the point at which travel into the cartridge case ends. Some brands 

consistently produce these marks whereas others don’t. But obviously these 

9

marks have a tendency to be over marked as the bullet makes its way down the 

barrel during firing. The questioned cartridge case should be placed on base 

down on the pad of leather or rubber to prevent damage to the toolmarks 

imparted during the previous firing event. A reload bullet must be placed nose 

first into the cartridge case. The bullet can now be tapped into the cartridge case. 

After indexing, use an inertia bullet puller to extract the bullet from the cartridge 

case. Bennett wrote a case report on identifying a bullet to a cartridge case (22). 

Striae can originate from case mouth burrs but it will depend on the relative 

hardness of bullet jacket versus cartridge case material if markings are 

transferred. Also Barnes produced a case report on the same subject (23). In a 

presumably cover-up murder case, it became necessary to evaluate cartridge 

case mouth crimp impressions on fired bullets. The easiest way was to compare 

castings from the inside of the case mouth directly to the fired bullets. In a case 

report, Clow looks at identifying bullets and cartridge cases to a single unknown 

firearm using magazine marks (24). Upon examination of cartridge cases and 

bullets from a homicide scene, it became apparent that both bear gross 

markings. The markings on the cartridge case were determined to be magazine 

markings. They were very distinct and identifiable and some cartridge cases 

even had multiple markings. At the stage were the bullets were compared, it was 

noted that the gross markings on bullets and cartridge cases could be positively 

inter-compared. Also the sequence of the multiple markings was in good 

correspondence, leading to the conclusion that both bullets and cartridge cases 

could be identified to the same gun. 

Siso and Kasachesko too report on identifiable magazine marks on the base of 

cartridge cases, fired from a FN HP pistol (25). It was discovered that the 

markings originate from the back top area of the magazine. The striated mark’s 

appearance depends directly on the shape and inside surface at the rear of that 

area. Also, the method of loading has an influence on the appearance of the 

markings. 

Koffman and Hocherman introduce class and individual chamber impression 

markings on cartridge cases fired by FN1910/1922 pistols (26). These 

characteristics, produced during the discharge process, can be found in three 

areas on the forward section of the rim. As class characteristics they can assist 

the firearms examiner to identify the type of firearm used. As individual 

characteristics they can be used as an excellent source for microscopic 

comparison examinations. 

2.1.4 Proficiency testing 

ENFSI send round a 2 nd edition of their proficiency test. The FAID2009, 

distributed early 2010, consisted of 10 sets of bullets and cartridge cases. Each 

set consisted of 2 known reference elements and third questioned element that 

had to be compared to the references. The proficiency test was made from 

castings (see also reference (55)). 

10

Collaborative Testing Services, Inc. remains well known as provider for 

interlaboratory tests. Tests can be ordered from their website and reports from 

past tests can be downloaded there too (27). 

2.1.5 Instrumental methods 

i. Automation and 2D/3D Imaging 

In the last years, there has been a noticeable increase on the market of 

companies that offer automated systems to aid the examiner in his comparisons 

with the vast amount of exhibits in the open case file. At the annual meeting of 

the ENFSI Expert Working Group on Firearms, and for the first time to our 

knowledge, were present no less than 6 manufacturers/providers of Automated 

Ballistic Imaging Systems: Forensic Technology Inc., Laboratory Imaging s.r.o., 

Papillon ZAO, Pyramidal Technologies, SBC and ScannBI Technology. They 

were given the opportunity to present and expose their products. 

In their March 2008 report, the National Academy of Sciences assesses the 

feasibility, accuracy, and technical capability of a national ballistics database 

(28). An executive summary of the extensive report is available free of charge. 

The main goal of the report was to study ballistics imaging technologies. 

However, three concerns were raised as to the scientific validity of firearms 

identification: 1/ whether or not the validity of the fundamental assumptions of 

uniqueness and reproducibility of firearm related toolmarks has been 

demonstrated; 2/ whether additional general research on the uniqueness and 

reproducibility of firearm related toolmarks should be done; and 3/ whether 

conclusions drawn in firearms identifications should be stated to imply the 

presence of a firm statistical basis. The Committee for the Advancement of the 

Science of Firearm and Tool Mark Identification of the Association of Firearms 

and Tool Marks Examiners (AFTE) has responded to those three concerns (29). 

Brinck has compared the performance of IBIS and BulletTRAX-3D Technology 

using .45 bullets (both jacketed and lead) fired through 10 consecutively rifled 

barrels (30). The study assessed the ability of both systems to correctly identify a 

hit and also where in the correlation list that hit lies. Also the quality of the images 

was assessed. As for the jacketed bullets both systems perform very well with 

the BulletTRAX-3D being slightly better. However, for the lead bullets, the IBISsystem 

could only correlate 30% of the hits in the first 20 positions whereas the 

BulletTRAX-3D correlates 100%. It was found that the image quality was greatly 

increased when the 3D images were compared. Also for the inter-composition 

comparisons, the BulletTRAX-3D demonstrated to be much more successful in 

finding the correct correlations. 

11

In (31) Sakarya et al. discuss the use of photometric stereo for reconstruction of 

3D cartridge case surfaces. In this paper the 3D information is obtained from 

multiple 2D images which are acquired using a specially designed set-up. The 

authors further describe the radiometrical calibration, photometric stereo, and 

surface fitting methods used for translating the 2D information into a 3D surface. 

The resulting method is low cost (as compared to e.g. laser interferometry or 

profilometry) and not time-consuming. Unfortunately, no comparison of the 

accuracy of this method, nor its relevance for performing real-life visual or 

automated matching in case work or database searching is included. 

Recent advances in 3D tomography techniques have resulted in multiple authors 

examining the use of these methods for studying incidents in which ballistic 

projectiles were involved. In (32) the use of cone-beam computed tomography 

(CBCT) is compared to multi-slice computed tomography MSCT/MDCT. The 

authors conclude that CBCT can be an important diagnostic tool for the 

evaluation of gunshot injuries, especially for detecting structural hard-tissue 

damage around or nearby projectiles, and for determining the precise location of 

a projectile in a body. Thali et al. (33) applied 3D MSCT in a case concerning an 

illegal lynx shooting and argue that it is a suitable and promising new technique 

for veterinary pathology. Puentes et al. (34) used 3D MSCT in a non-fatal case 

where a man was shot in a multiple aggressor situation. They used this method 

to accurately determine the bullet’s trajectory. In (35) a CT scanner is used to 

analyze the cavities and the fragmentation and dispersal pattern of the projectiles 

that occurred after discharging a firearm in commercially purchased ballistic 

glycerine soap blocks. The authors argue that this method offers a fast and 

accurate analysis that also avoids issues such as e.g., the possible drying or 

shape alteration of a block that can occur when using a traditional examination 

process. 

De Smet et al. (36) discuss the use of both traditional optical and 3D measuring 

techniques to investigate if the initial seating depth of the primer has an impact 

on the variability of the marks found on the breech face. 

They conclude that for the circumstances under which they tested (i) no clear 

impact on the primer striation patterns could be found, but that (ii) differences in 

the firing pin impressions could be noted. In this work 3D measuring did not yield 

important additional information for arriving at these conclusions. 

Song and Vorburger continue their earlier work on topography measurement 

techniques. In (37) they propose a new “profile difference” parameter for 

measurement and comparison of 2D and 3D surface topologies. They also 

studied the topography decays and the impact of the electro-forming technique 

used by the National Institute of Standards (NIST) to produce their standard 

reference material (38). They conclude that for the 256 casings under 

consideration it can be expected that they are replicated with a proposed 

CCFmax correlation rate of higher than 95%. 

12

Chu et al. realized a pilot study for automated topography and correlation based 

bullet signature identification. They examined 48 bullets fired from six different 

barrel manufacturers. First, they classified these bullets into groups based on the 

LEA (land engraved area) width class characteristic. Next, they used the crosscorrelation 

function to determine the effective correlation area, and extracted 

each 2D bullet profile signature. The list of top ranking database candidates of 

bullets fired from the same model firearm was then obtained based on the crosscorrelation 

maximum values. They report a 9.3 % higher accuracy rate compared 

to a currently used commercial system (39), and conclude that by following the 

proposed sequence of methods the correlation results can thus be improved. 

In (40), Xie et al. also present 3D topography methods for bullet marks and 

characteristics extraction. They extend their proposed methods with a variety of 

2D and 3D visualization graphics that can help firearm examiners in their work. 

Xiao-ning et al. discuss a certain relationship between texture features and the 

erosion and wear images in the artillery chamber as observed during the useful 

life of a gun (41). More specifically, the relationship between the texture features 

and the total round number is studied using an improved algorithm based on the 

Differential Box Count method. 

In (42) the authors discuss the impact of using different illumination wavelengths 

(400 nm - 700 nm) on the quality of the obtained projectile images. The authors 

conclude that longer wavelengths yield better images which are important as 

high quality images are needed for accurate comparison and identification. 

Feng et al. present their work (43) on the scale invariant feature transform (SIFT) 

and graph transformation matching for building stitched mosaics of cartridge 

case images. In the fusion stage they apply an additional algorithm to smooth the 

seems between images. 

Ghani et al. discuss the use of geometric imaging moments for firearm 

identification (44). They analyzed 747 cartridge case images captured from five 

different pistols using 48 initial features which were analyzed using a MANOVA 

test. Using correlation analysis they further reduced this high dimension of 

features into 11 significant features after which cross-validation under 

discriminant analysis results in 96.7% of the images being classified correctly. Ali 

et al. propose novel computer vision based methods for automatically scoring 

shooting targets (45). They report a resulting bullet count accuracy of 98.3% for 

the proposed algorithm after testing on 100 target images with a varying number 

of bullets hit. 

For building an automatic system for firearm identification, Thumwarin discusses 

the use of Fourier coefficients of cartridge case primers with a polar coordinate 

system, a finite impulse response system and Fisher's linear discriminant 

function to identify the gun (46,47). 

13

Li et al. also use the Fast Fourier Transform (FFT) for analyzing the line-scan 

imaging technique of projectiles (48). In (49) the line-scan technique is compared 

with laser depth proofing and photonic 3D topography. 

Blechko and Darker et al. discuss the ability of lay and experienced operators, 

and an automated vision method to detect an individual carrying a (concealed) 

firearm in simulated CCTV footage (closed circuit television); see e.g. (50,51,52). 

They conclude that although several issues remain to be investigated or 

improved further: (i) a human observation inspired method would most likely be 

the most successful automated method, (ii) a combined automated and human 

approach may offer potential for realizing the given detection task in real-life, and 

(iii), the decision making process whether a person is carrying a concealed 

firearm appears to be linked to the emotional state of the observed person. 

Chu et al. studied striation densities in images obtained for six lands of 48 bullets 

fired from 12 gun barrels of six manufactures (53). They conclude that their 

statistical results show a strong support for a relationship between identification 

rate and striation density. The authors offer that their method can thus provide a 

quantitative criterion to assess whether sufficient striae are present in order to 

achieve reliable bullet identification. 

ii. Equipment 

Katterwe et al. introduce comparison scanning electron microscopy in forensic 

science and demonstrate the applications for firearms (and tool mark) examiners 

(54). The comparison VP-SEM (Variable Pressure – Scanning Electron 

Microscopy) consists of two separate SEM that are linked so that an image of the 

second can be displayed alongside an image of the first. Due to its high 

resolution and depth of field, the system allows for detailed comparison of 

specimens. Examples are manifold: cartridge cases and bullets but also cut 

adhesive tapes and other applications. Samples can be either conductive or nonconductive. 

As it comes to harmonising results from proficiency tests, it is important that all 

participants are looking at the exact same samples. For this reason an advanced 

casting technique was developed by Koch and Katterwe (55). In their article they 

present the “3D-mass-produced casting procedure”, including the construction 

and manufacturing of the used devices. 

2.1.6 Court rulings 

Particularly the firearms and tool marks examiners in the USA are regularly 

challenged in court to defend the scientific principles of the discipline. The 

Defense will often motion for a Frye or Daubert hearing to exclude the evidence. 

The Scientific Working Group Firearms (SWGGUN) is keeping track of these 

14

court rulings on their website (56). The following trials may be of interest to 

firearms examiners: 

� State of Maryland v. Henry Wittingham # 08-1682X, December 30, 2009; 

Circuit Court of Prince George’s County, Maryland (Judge Nicholas E. 

Rattal) 

� State of Ohio v. Demonte Merriweather # 514703, October 30, 2009; 

Court of Common Pleas (Judge Jose A. Villanueva) 

� U.S. v. Tavon Mouzone # WDQ-08-086, October 29, 2009; U.S. District 

Court for the District of Maryland (Judge Paul W. Grimm) 

� State of California v. Earl Eugene Rose, No FVA 010494, September 2, 

2009, Superior Court of California, County of San Bernardino (Judge 

Michael A. Smith) 

� U.S. v. Joseph Thomas #2008 CF1 25845, September 4, 2009, Superior 

Court of the District of Columbia (Judge Herbert B. Dixon) 

� U.S. v. Donald Scott Taylor, Case # 2:07-CR-01244 WJ, September 30, 

2009, District Court of New Mexico (Judge William Johnson) 

� U.S. v. Willie Gayden, Criminal Case #2006 CF1-27899, June 26, 2009, 

Superior Court of the District of Columbia (Judge Michael L. Rankin) 

� State of Indiana v. Desmond Turner, #49G02-0606-MR-101336, June 17, 

2009, Marion Superior Court (Judge Robert R. Altice) 

� State of Alaska v. Derrick Wren, Case #3AN-03-10649CR, May 1, 2009, 

Superior Court, Third Judicial District at Anchorage (Judge Michael 

Spaan) 

� State of Ohio v. Eashawn Anderson, Case #CR509503, March 25, 2009, 

State of Ohio, County of Cuyahoga, Court of Common Pleas (Judge 

Michael J. Russo) 

� Commonwealth of Pennsylvania v. Steven Carl Colegrove, CP-08-CR- 

0000785-2007, January 15, 2009, Court of Common Pleas, Bradford 

County, PA (Judge Maureen T. Beirne) 

� U.S. v. Chaz Glynn, Case # 06 CR 580(JSR), September 2, 2008 and 

U.S. v. Damian Brown, Case # 05 CR 538, June 30, 2008, District Court of 

New York, Southern District (Judge Jed S. Rakoff) 

� U.S. v. Troy Worsley, 2003 FEL 6856, June 2, 2008 and U.S. v. Kevin 

Edwards Case #F-516-01, Superior Court of the District of Columbia 

(Judge Neal E. Kravitz) 

� State of Washington v. Tony Smith, No. 03-1-00045-7 KNT, August 15, 

2008, State Superior Court for King County (Judge George T. Mattson) 

� U.S. v. Ronald English, Criminal #2007 CF1 16618, March 12, 2008, 

Superior District Court of the District of Columbia (Judge Geoffrey M. 

Alprin) 

� State of Connecticut v. Robert A. Legnani #AC 26840, February 5, 2008, 

Superior Court of New Britain, Connecticut (Judge J. Cronan) 

� U.S. v. Khalid Barnes, Criminal #S9-04-CR. 186, April 2, 2008, Southern 

District of New York (Judge Stephen C. Robinson) 

15

� Maryland v. Monti M. Fleming, Criminal Case #06-46624, February 13, 

2008, Circuit Court of Howard County (Judge Louis A. Becker) 

2.2 Firearms & ammunition miscellaneous reports 

2.2.1 Firearms 

Thomas Warlow has written an extensive manuscript on the criminal use of 

improvised and re-activated firearms in Great Britain and Northern Ireland (57). It 

shows that after introduction of a firearm ban, criminals find other ways to import 

firearms and distribute them on the market. The converted and re-activated 

firearms show very distinct markings although some of them have smooth bores, 

resulting in the recovery of unrifled bullets at the crime scene. As time went by, 

the re-activation techniques became more and more professional. 

Hannam publishes on the firearm trends in England & Wales (58). The 

information was obtained from the National Firearms Forensic Intelligence 

Database (NFFID) and covers the time period 2003-2008. It was concluded that 

21% of the annual submissions, were converted weapons. The makes, models 

and modes of conversion of these weapons are described in detail. Yilmaz et al. 

report on handmade guns in Trabzon, Turkey (59). Although very similar in look 

to commercially available weapons, these guns are constructed from cheap 

materials and are not safe for firing. 

Sarıbay, Hannam and Tarımcı investigated the legal status of modified shotguns 

in Turkey (60). Over the period 2006 to 2008 11% of all seized shotguns were 

converted. According to Turkish law these are either considered as a pistol or as 

a nonstandard shotgun. 

2.2.2 Ammunition 

Bailey has developed a classification system and identification key for .177 

calibre pellets (61), based on a five-class characteristics criterion: pellet head 

shape, skirt type, pellet length, weight, markings and other observations. This is 

not an absolute pellet identification system; however, it provides possible product 

identification of some pellets for the investigator. Also, the system can eliminate 

numerous pellet brands. 

2.2.3 Recovery of obliterated serial numbers 

It can be a challenge to recover laser engraved serial numbers that have been 

obliterated. This is true because the underlying principle is different from a 

stamped serial number. Da Silva and dos Santos (62) present a case report on 

an aluminium alloy pistol. The trick is to polish carefully and slowly and to capture 

subsequent images with a camera. One of the major drawbacks of this method is 

that it is time consuming. Klees looked into the possibility of using SEM and X- 

16

ay mapping to restore or detect obliterated laser-etched firearm markings (63). It 

was concluded that the use of SEM and X-ray mapping as a non-destructive 

method to restore or detect totally obliterated laser-etched markings was 

ineffective. 

A paper by Wightman and Matthew (64) looks at the application of Fry’s reagent 

to recover erased marks from steel. The novelty of this publication is that it looks 

into the effect of the original stamp force on the ability to recover erased marks. 

The applied force is important since this controls the degree of underlying 

deformation as well as the depth of penetration. Recovery of erased marks is 

difficult where low applied force has occurred. In the present study marks could 

still be recovered 1.25 mm below the impression. Also the authors have noted a 

degrading effect of the Fry’s reagent in function of storage time. Nowadays more 

applications use high strength aluminium alloy. When engraved marks are 

obliterated from such substrates, current techniques prove unsatisfactory. Bong 

and Kuppuswamy have investigated what etching techniques are better in these 

situations (65). It appeared that either 1/ immersion in 10% aq. phosphoric acid 

(H3PO4) and 2/ alternate swabbing of 60% HCl and 40% NaOH were both quite 

effective. 

2.3 Legislation and other efforts 

2.3.1 Microstamping 

In the past years, a lot of debate has been going on regarding microstamping of 

codes or serial numbers on firearm parts, more specifically the working surfaces 

such as the firing pin, that are transferred to the cartridge case during the 

discharge of the gun. With the AB 1471 Crime Gun Identification Act of 2007, 

microstamping became legislation for the state of California. Equivalently, 

Assembly Bill AB 09818 passed New York legislature. In those states and as of 

January 1, 2010 semi-automatic pistols need to be equipped with a microscopic 

array of characters that identify the make, model, and serial number of the pistol. 

It appears that serious testing by qualified firearms examiners was negated. In 

an open letter to the AFTE membership, Lucien C. Haag comments on the false 

claims that were made regarding his early study of the microstamping technology 

(66). 

In 2008, Beddow, Howitt, Tulleners and Giusto published an extensive study on 

the durability and legibility of micro-markings from firing pins (67). This second 

generation of micro-engraved firing pin contains three different encoding formats: 

1/ the original alphanumeric code on the face of the firing pin, 2/ either a gear 

code or a dot code, and 3/ a radial bar code that extends down the side of the 

pin. Overall the alphanumeric characters and the gear code structures proved to 

be capable of withstanding repeated firing; however some degradation was seen 

with specific firearms. A determination of what constitutes an acceptable lifespan 

for these characters needs to be developed. The dot code structure was 

determined to be unsuitable. The radial bar code structure exhibited varying 

17

degrees of degradation with specific occurrences of complete obliteration. The 

transfer rate of the micro-markings was clearly dependent on the brand of 

ammunition. Also, the encoding structures on the firing pin can be damaged or 

obliterated with relative ease. 

2.3.2 Glock EBIS barrel 

It is usually extremely time-consuming and not always possible to identify bullets 

that have been fired through polygonal barrels. As many such firearms, e.g. 

Glock pistols are used by law enforcement, this has triggered a research project 

to enhance the identification capabilities. One of Glock’s attempts was the 

Enhanced Bullet Identification System (EBIS). Martinez did an extensive study 

looking into the durability of the EBIS barrel markings after 250 – 10000 rounds 

(68). The EBIS signature imparts gross lines down the length of the barrel on the 

lands of the rifling, like a barcode. Results of the study show that these gross 

lines endure but that there was a disparity with the finer lines that reduce the 

chance of identifying Glock bullets by a sufficient correspondence of individual 

characteristics. 

2.3.3 Bore lapping 

In a feasibility study, Northcutt wanted to determine if modification of polygonally 

rifled Glock barrels using Permatex valve-grinding compound 34A would facilitate 

the forensic identification of fired bullets (69). Indeed it was found that 

modification increases the number of reproducible individual markings present on 

fired bullets and allow for identifications even after 900 shots were fired. As a 

side note, the accuracy and muzzle velocity of modified barrels were compared 

to the unmodified barrels. No significant differences were observed. 

Haag et al. have examined a series of bore-lapping compounds to study their 

effect on firearms identification (70). Although these compounds were developed 

to improve the exterior ballistic performance of firearms, they were generally 

found to alter the bearing surfaces within the firearm’s bore to the extent that pretreatment 

and post-treatment bullets could not be identified to each other in all 

cases. However, “new” patterns of striae occurred on the land impression that 

are sufficiently persistent that post-treatment bullets could be identified among 

themselves and often possessed striae patterns that were more pronounced than 

those on pre-treatment bullets. Such treatments will not be obvious from an 

inspection or casting of the bore nor is SEM-EDX (Scanning Electron Microscopy 

– Energy Dispersive X-ray microanalysis) analysis of the bore’s surface likely to 

detect residues from such treatment methods. 

18

2.4 Shooting Incident Reconstruction 

2.4.1 Research 

For those individuals that are doing trajectory reconstructions, it is interesting to 

know how accurate and precise the measurements of the angles are. Michael 

Haag summarises data that was collected over many years (71). As for the 

precision it was concluded that 6 degrees would yield a confidence level of 99%. 

Equivalently for the accuracy it was concluded that 5.1 degrees yields a 

confidence level of 99%. The author furthermore talks about the newest 

technique which is 3D laser scanning of the crime scene. 

Haag et al. have also published an extensive study regarding ejection patterning 

from a series of pistols (72). The article discusses standard protocols for testing 

as well as the effects the firearm’s mechanism, pitch, rotation, and motion have 

on the final rest locations of fired cartridge casings. Typically pointing the muzzle 

of a firearm downward will move the pattern of casings forward of the firearm’s 

location, while pointing the muzzle upward may increase or reduce the rearward 

component of a pattern. Also motion has a tremendous influence as it adds an 

extra moving vector component to the ejected case. It was concluded that 

ejection patterning can give an estimation of the shooter’s position but it is not a 

high-resolution tool. Preferably ejection patterning is coupled with other aspects 

of shooting reconstruction to increase the accuracy of the placement of the 

firearm. 

Haag reports on muzzle flash duration and the factors dictating its presence or 

absence (73). It was observed that contemporary handguns and ammunition can 

produce noticeable and visible muzzle flash particularly when discharged in low 

light conditions. The duration of the visible discharges can be very short (far less 

than 1/60 of a second). This is substantially less than the time duration of a 

spontaneous blinking of the eyes; consequently it is quite possible that a witness 

who spontaneously blinked at the instant a firearm was discharged would not see 

the muzzle flash. 

Tsach et al. have developed a model to estimate projectile impact velocity on 

metal sheet targets from the shape of the target hole (74). Experiments were 

performed with the 5.56 mm calibre fired from a M-16A1 assault rifle. A paper by 

Rezos et al. gives insight in the maximum burial depth that hidden firearms could 

still be recovered using a basic all-metal detector (75). It appears that the size of 

the weapon and surface area were the two variables that affected maximum 

depth of detection. The maximum depth for detection was 40 cm or less although 

with high settings and for the largest firearms, one could detect up to 55 cm. 

19

2.4.2 Case reports 

Bresson and Franck (76) report on a long distance shooting for which they 

determined the shooting distance in a somewhat particular way. In their 

experiment they fired bullets at different velocities to a concrete plate. The bullet 

damage was consequently compared to the damage observed on the crime 

scene bullet. The damage was quantified by length and diameter. The velocity 

that was calculated from these experiments was used to extrapolate to the 

shooting distance. The greatest degree of uncertainty in this method arises from 

the lack of information of the muzzle velocity. Hu et al. present a case report 

where SEM/EDX was used to reconstruct the shooting trajectory. It demonstrates 

the evidential value of trace analysis (77). 

2.4.3 Trace analysis 

Wehner et al. (78) have investigated the possibility of associating bullets found at 

the crime scene to specific trajectories in the body of a gunshot victim. They 

investigate heart- and liver-specific cell material on the bullets. The method is 

particularly useful in the case of expanding bullets where the insides of those 

bullets can be swabbed. 

Goddard et al. use advanced methods to examine latent fingerprints on brass 

surfaces (79). As brass is a very common material in ammunition, this can be of 

interest. Atomic Force Microscopy (AFM) is used to visualize corrosion of the 

metal on a nanometre scale. The method shows in detail the corrosion 

mechanism between the fingerprint deposit and the metal after heat treatment of 

the latter. Also Bond has done quite some work on fingerprint evidence. In a 

publication he goes into depth on the visualization of latent fingerprint corrosion 

of metallic surfaces (80). It was established that the salts in the fingerprint 

residue corrode the metal surface to produce an image of the fingerprint that is 

both durable and resistant to cleaning of the metal. The author proposes a heat 

treatment to enhance the durability of the fingerprint. Also cartridge cases need 

to be heated additionally after discharge. The return, however, is low as cartridge 

cases tend to lose some of the fingerprint evidence during loading and ejection. 

For these types of evidence, the author suggests an electrostatic enhancement 

technique so the conducting powder adheres better to the corroded areas. The 

same author has continued this work in another publication (81). Here is 

presented a case report in which the author recovers an old latent fingerprint on 

a brass cartridge case. It demonstrates that novel techniques can visualize 

fingerprints where cyanoacrylate fuming can fail to do so. Edmiston and Johnson 

propose an optimal sequence for chemical development of latent prints on 

cartridge casings and shotgun shells (82). 

Gashi et al. have examined, using infrared video evidence, the temperature 

developed during discharge of 9 mm cartridge cases (83). It was concluded that 

20

it should not affect any DNA residues left on the case. In other words: DNA 

sampling of spent cartridge cases is encouraged. 

2.5 Wound Ballistics 

Over the last three years quite a number of articles have been published on the 

subject of injuries caused by firearms. Also a few reviews were written containing 

statistical information regarding firearm related deaths: are they homicides, 

suicides or rather accidents, what type of firearms are used, how many gunshot 

wounds were inflicted… Although this statistical information is perhaps not 

directly related to the technical field of ballistics, it can be interesting background 

information for those who are regularly examining firearm fatalities. 

2.5.1 Statistics 

Solarino, Nicoletti and Di Vella (84) discuss the situation between the years 1988 

and 2003 in the city of Bari in the South of Italy. In this area of the country, nonnatural 

deaths are mainly caused by firearms. The data that is collected in this 

review paper is from the autopsy reports from 717 firearm-related deaths: 634 

homicides (88.4%), 82 suicides (11.5%) and 1 accidental. It appears that most of 

the victims were males (93 %) with an average age of 25.6 years (females have 

an average age of 31.3 years). The average age of a male committing suicide is 

40.3 years. Only one female suicide case was reported. Pistols were the most 

frequently used firearm (46% for both homicides and suicides). Together with the 

revolvers they account for 54% of the homicides and 48% of the suicides. The 

most commonly used calibres for homicides are the 7.65 mm and the 9 mm. The 

9 mm is also the main calibre for suicides. In homicides multiple gunshot wounds 

are usually reported (65.2% of the homicide cases). 4.8% of the suicide cases 

involved two gunshot wounds but no more than that. Two of those cases involved 

entrance wounds in the limbs. In homicides most wounds are inflicted to the head 

(23.6%), the chest (20.9%), the back of the trunk (16.9%) and the abdomen 

(10.6%). In suicides the percentage of entrance wounds in the head is much 

higher (67.4%). Interesting is the anatomical distribution to the head (right 

temple: 46.5%; left temple 23.3%), mouth (10.3%) and middle forehead (10.3%). 

There is also a direct relation between the location of the entrance wound and 

the type of firearm used: handgun to the temples, shotgun to the chest, abdomen 

or mouth. The review paper further discusses reasons and motives for suicides 

and homicides and compares with other types of homicide and also with 

information from other countries. 

Verzeletti, Astorri and De Ferrari (85) prepared a comparable study in 2009 on 

the situation in Northern Italy between the years 1994 and 2006. In this period 

164 fatalities were identified as being firearm-related. From this number 59 

(35.9%) were homicides and 99 (60.4%) were suicides. 3.7% were accidents. 

This is a different situation from the South of Italy (see reference (84)). Undoubtly 

the demographic situation plays an important role. Most suicides seem to take 

21

place in winter and spring but for homicides there is no specific monthly 

predominance. As it pertains to the gender, 72.9% of the homicide victims were 

male and in the suicide group 92.9% were males. The mean age of a homicide 

victim was 37.9 years and for a suicide victim was 47.2 years. Those ages are 

somewhat higher than what was observed in Bari. As for the type of firearms, 

single action weapons accounted for 68.3% of all deaths whereas multiple action 

guns only played a role in 30.5% of the cases. Most of the injuries (suicide and 

homicide) are located on the head (38.6%), the chest (29.3%), the neck (13.4%), 

the abdomen (9.3%) and the limbs (9.3%). In suicide cases the anatomical 

distribution is as follows: right temple (34.4%), the precordium (20 %), the neck 

(16.7%) and the mouth (7.8%). Multiple wounds (25.2% of the cases) are more 

common for homicides (57.9%) then for suicides (6%). The authors have also 

compared to a previous study in the same geographical area. Apart from a rise in 

the number of suicides, the other data stayed relatively equivalent. A series of 

sketches nicely illustrates the distribution of the entrance wounds in the cases of 

homicide, suicide or accident. As a side note it is interesting to observe the 

toxicological results on the victims. The authors furthermore compare the 

situation in Brescia to other areas in Italy and on the international level. One 

should carefully take into account all cultural and regulatory aspects to be able to 

compare different situations but a clear correlation is demonstrated between 

social factors, poverty, crime and drug use on the one hand and access to 

firearms on the other hand. Gill and Pasquale-Styles give statistical information 

regarding firearm deaths by law enforcement over a 4-year period in New York 

City (86). Byard et al. compare the characteristic features of paediatric firearm 

fatalities between Australia and the United States (87). 

Another study by Balci, Canogullari and Ulupinar (88) presents the 

characteristics of gunshot suicides. In gunshot fatalities, to determine whether 

the cause of death originated by suicide the following information is particularly 

valuable: the shooting distance, firearm discharge residues on the hand, the 

number and localisation of entrance wounds. The statistical analysis 

demonstrates a significant age difference for people committing suicide by 

firearm (29.2 ± 12.8 years) and other means (39.7 ± 17.8 years). The percentage 

of gunshot suicides decreases as age increases. The percentage of short 

firearms is quite comparable to the previously cited studies. The authors 

observed a relatively high number of suicides (27.7%) with a shooting distance 

from intermediate range around arm reaching distance. A detailed table shows 

the relation to the anatomical position of the entrance wounds and the firearm 

that was used (either short or long barrelled). 6.1% of the cases showed multiple 

entrance wounds. It appeared that the victims were all soldiers with their rifles set 

to automatic. The authors conclude that access to weapons plays an important 

role in the prevalence of the gunshot suicides. 

Also Lorin de la Grandmaison, Fermanian, Aegerter and Durigon have published 

a retrospective study, this one regarding the use of long firearms (89). This study 

has especially focused on the usefulness of the projectile trajectory as an 

22

indicator of homicide vs. suicide. In depth statistical analysis was performed on 

correlation between homicide/suicide and gender, age, number of shots, range of 

fire and direction of the projectiles. Furthermore, the article contains a number of 

references to e.g. location of the firearm after suicide or for discriminating 

between suicidal or homicidal injuries. Those references can be of interest for 

those examining suicides by firearm. Although all of the above studies were not 

inter-compared in-depth, information regarding age, gender, number of shots, 

range of fire and location of the entrance wounds is consistent. 

Firearms legislation has clearly an influence on the suicide by firearm rate. 

Sarma, Griffin and Kola (90) discuss the situation in Ireland as it comes to access 

to firearms by people suffering from mental illnesses. Over a period of 23 years 

(1980-2003) a total of 725 firearm-assisted suicides occurred. Male victims of 

suicides were statistically more likely to have used a firearm than female victims. 

Most of these suicides occurred between the ages of 15 and 44. Also firearmsassisted 

suicides are more common for residents of rural areas. The international 

experience is that restrictive policies on firearms ownership and access can lead 

to decreases in levels of gun-related deaths and injuries. Compulsory safe 

storage reduces access to firearms by both criminal elements and members of 

the certificate-holders immediate family. 

Continuing their analysis of homicides and suicides in Bexar County (TX), Molina 

et al. (91) presented a long-period study of the trend of firearms usage in 

homicide and suicide focussing on the calibre of the used firearm. About 60% of 

the homicides and suicides are committed by firearms - mostly hand guns. 

Although there is a steady decline in the use of firearms in homicides (and 

homicides in general) since 1994, the use of firearms in suicides remains more or 

less a constant. Regarding the calibre of the handgun used, there is an increase 

in the use of medium and large calibre weapons, both in homicide and - to a 

lesser extent - in suicide cases. This probably reflects the availability of these 

types of weapons on the market for homicides, while suicides are mostly 

committed with a weapon already in possession by the shooter, which does 

therefore not immediately reflect market trends. Hejna and Šafr published some 

data regarding presence / absence of shots through clothing in firearm suicide 

cases (92). It was concluded that only in 9% the clothing was removed by the 

victim (e.g. shots through the chest). 

2.5.2 Case reports 

Besides the review papers cited above, a great number of case reports regarding 

gunshot wounds were published. Frank et al. (93) report on vole captive bolt 

devices, which are special mouse traps that belong to the spring guns. The 

ammunition is the 9 x 17 cartridge. The vole is killed by a high-pressure gas jet. 

In this paper, the authors discuss accidental hand injuries which range from soft 

tissue damage to the separation of a hand. 

23

Simic et al. report on the characteristics of head wounds, inflicted by captive-bolt 

gun, that are used for humane slaughtering (94). The authors compared 

experimental studies with results from autopsies of (usually) suicide victims. The 

wounds typically consist of a central round defect of the skin and underlying 

tissue. The edges of the wound entrance are flat and smooth. In the close 

proximity of the skin defects there are two symmetrically arranged eccentric oval 

zones of blackening. If the device was put at an angle with respect to the skin, it 

could be that the skin was only cut semi-circularly. In these cases, the blackened 

zones tend to have a wider and paler appearance. 

Chowaniec et al. (95) present a case of a fatal gunshot from rubber projectiles. In 

this case, the ammunition being used by the police officers was 12 x 70 

cartridges containing different rubber objects: 6 mm balls and a 34 x 14mm sluglike 

projectile. These types of projectiles were also recovered from the deceased 

that showed a severe injury to the heart. During a ballistic test the event was 

reconstructed with the so called Rój and Chrabąszcz cartridges that were also 

used on the scene of the crime. It was concluded that this ammunition, when 

fired from a short distance, can account for the observed woundings. 

Fracasso, Löhrer and Karger (96) describe a case where a woman self-inflicted a 

wound with an 8 mm Blank pistol in an attempt to simulate a criminal offence. A 

typical near-contact entrance wound was observed as was a large stellate exit 

wound. 

Hagemeier, Schyma and Madea (97) report a homicide and suicide by stud gun 

case, which is an explosive, cartridge-activated anaesthetisation device. At 

autopsy the characteristic signs of bolt shot injuries were identified: a round, 

punched defect of the skin and the cranial bone, bone fragments in the bold shot 

canal as well as a destruction zone longer than the depth of bolt penetration in 

the brain. Typically such devices have powder exhausts that lead to a radialsymmetrical 

powder dispersal that can be found around the entrance wound. The 

device in the present case, however, had no such powder exhausts. 

Two cases of suicide by unmodified blank pistols are discussed in a case report 

by Buyuk et al. (98). The authors have observed two cases where a shot being 

fired at close contact range caused gas (and bone) to penetrate into the body. 

The entrance wounds are relatively big and irregular and surrounded by soot. 

The extensive wound cavity is filled with powder particles and soot. The victims 

died of brain haemorrhage and brain tissue damage. 

Hejna and Šafr (99) published a case report on suicide by zip gun or improvised 

firearm. After first been overlooked by the investigators, a gunshot wound was 

discovered during the victim’s autopsy and a 7.65 mm bullet was recovered from 

the head. A subsequent re-investigation of the crime scene pointed to a device 

that was used by the victim as an improvised firearm. 

24

Another case report was published by Cunliffe and Denton (100). A victim was 

found dead with an atypical gunshot wound of the head. At the first examination 

of the crime scene, the gun was overlooked and the hypothesis was homicide. 

As no rifling marks were found on the bullet that was removed from the skull, a 

further police investigation was requested. A home made gun was discovered, 

which consisted of a 2-inch length of pipe, a 1 ¾ inch connector, an air hose 

nozzle and a 2-inch nail. A .38 Special cartridge case was found embedded 

within the pipe. 

Linert et al. report on an unusual case of suicide employing a shotgun barrel, a 

shotgun shell and a BB (101). The case serves as another example of the 

importance of cooperation between scene investigators and forensic 

pathologists. A review of suicidal firearm cases using unconventional means 

and/or firearms is also presented. 

Üzün et al. examined fatalities caused by spherical bullets fired from blank 

cartridge guns in Istanbul, Turkey (102). 59 fatalities had been reported where 

blank firearms were used that appeared to have been easily modified. It was 

possible to successfully fire 4 or 5 mm diameter objects. 

As for regular firearms, Lepik et al. (103) compared the injuries caused by 

Tokarev, Makarov and Glock 19 pistols at variable firing distances. In their study 

they also compare between cloth targets (cotton and synthetic) and human skin 

from autopsy material. At a shooting distance 25 cm clear soot deposit was 

visible. This pattern gradually disappeared from distances of 50 cm and higher. 

On the skin starting from a distance of 25 cm (Glock) and 50 cm (Tokarev and 

Makarov), no more soot was macroscopically observable. Also the authors have 

investigated microscopically the penetration of gunpowder residue particles in the 

skin, which is clearly depending on the firing distance and the shape of the 

individual powder grains. 

Kuchta and Klug (104) report on a rare case where a patient had several selfinflicted 

gunshot wounds to the head without fatal end. Three bullets were 

recovered from undeep entrance wounds. The gun that was used was a Second 

World War 7.65 mm Luger handgun. The ammunition being used was 

degenerated after storage of 50 years. 

2.5.3 Research 

Frank has continued his analysis of vole captive bolt devices together with a few 

other co-workers. In a research article (105) they measure the actual pressures 

that these devices release. It appears that the pressure in the explosion chamber 

goes up to 1100 bar and that the flow velocity of the gas jet was up to 2000 m/s. 

There is an inverse ratio between the maximum impact force of the blast wave 

and the distance to the muzzle. The force was up to 11500 N for the contact shot 

distance leading to an energy density far beyond what is required for skin 

penetration. For completeness, Frank has taken this another step further and 

25

measured the ballistic parameters of the 9 x 17 mm cartridge that is commonly 

used for such devices (106). 

In two different articles, Ogunc et al. (107) and Saribey et al. (108) examine the 

wounding potential of modified blank ammunition. Blank weapons are fairly 

accessible (Blank pistols costing only 10% of "real" guns can be purchased by 

over 18-year olds without any license) in Turkey and were seized in great 

numbers in modified condition. The authors discuss the modification process: 

either replacement of the original obstructed barrel with a smooth one, in which 

cases the firing of live ammunition becomes possible or the removal of the 

obstruction (restraint piece) in which case only modified blank ammunition can be 

discharged. In the latter case, the ammunition is adapted as follows: removal of 

plastic wad and replacement with a lead or steal sphere (shotgun pellets or ball 

bearings). The wounding potential was examined using ballistic gelatine and 

ballistic candle blocks, coated with a skin stimulant and different clothing 

simulants. In all cases, penetration occurs, it being deeper for the steel (mean: 

8.608 cm) than for the lead spheres (mean: 6.434 cm) (bare skin situations). It 

can be expected that fatal injuries can occur if such firearms are discharged from 

a close distance. These modified blank guns are more and more being 

encountered in criminal activities. 

As for wound ballistic testing, gelatine is a long established and reliable tissue 

simulant. The projectile penetrating the gelatine transfers energy and causes 

radial cracks according to the temporary cavity. Gelatine can be an ideal way to 

study the behaviour of air rifle pellets, as was done by Wightman and his 

colleagues (109). They have investigated the influence of quite a few parameters 

on the behaviour of the pellets: comparing 1/ different gels, 2/ different gel 

concentrations, 3/ different delivery powers, 4/ pellet shape, 5/ firing range. They 

have furthermore examined the effect of bone: damage to the bone was not 

observed but the pellets suffer important deformation. Computed tomography 

allows examination of bone and pellet damage in situ in 3D. In ballistics gelatine, 

the crack length is a function of the energy spent. Schyma used colour contrast 

(acryl paint) to render the fissures more visible (110). It was concluded that doing 

so, the measurements were more reproducible and the curve of the wound 

profile typically shifts by 1 cm to the entry. 

Frank et al. examined the trauma potential and ballistic parameters of cal. 9 mm 

P.A. dummy launchers (111). These are typically used for dog training: a hard 

foam dummy is launched for the dog to retrieve. It was concluded that for two 

projectiles (1/ the complete foam dummy and 2/ the aluminium mounting sleeve 

separately), the energy densities were 0.069 J/mm² and 0.282 J/mm² and thus 

high enough to cause relevant injuries. 

26

2.6 Training Material & Books 

Since 2008, the NFSTC put a firearms examiner training course on line (112). 

This course was made in collaboration with AFTE members and based on the 

AFTE training manual. Also recently released to the press was a new edition of 

Modern Scientific Evidence: The Law and Science of Expert Testimony. A 

chapter of this book is dedicated to Firearms and Toolmark Identification (113). 

Brian J. Heard published the second edition of his book entitled “Handbook of 

Firearms and Ballistics – Examining and Interpreting Forensic Evidence” (114). 

3 Firearms – Chemistry 

During discharge from a firearm, primer and gunpowder residues as well as 

metal particles from the projectile and the cartridge case are escaping from the 

muzzle and other openings of the firearm. These residues are referred as primer 

residue, firearm discharge residue or gunshot residue (GSR). 

3.1 Inorganic GSR 

Scanning Electron Microscopy – Energy Dispersive X-ray microanalyses 

(SEM/EDX) still is the method of choice for the identification of inorganic GSR on 

samples. This technique is well adapted to the detection of small particles 

containing heavy metals such a lead, barium and antimony coming from primer 

with a "conventional" composition. 

3.1.1 Lead-free ammunition 

However more and more primers do not contain one (or more) of these three 

elements. Because of the "uncharacteristic" composition of the residues, 

analysis of these types of ammunition by SEM/EDX will often be falsely negative. 

Fortunately, in reality the "memory effect" of the gun produces sufficient 

"contaminating" classic particles to make a good identification still possible. The 

use of special tagging compounds and metals in the cartridge is nevertheless 

highly recommended. Automatic analysis system parameters must be adapted 

since standard runs will miss these particles or yield inconclusive results. Taking 

note of these considerations, Martiny et al. (115) performed a thorough 

characterisation of the gunshot residues produced from Brazilian lead-free 

ammunition by SEM/EDX analysis. The "CleanRange" lead-free ammunition 

made by Brazilian CBC was originally developed considering health issues with 

lead-containing classic ammunition, for instance when used in police shooting 

ranges. They have since made their entry in the general population of 

ammunition used in Brazil. In fact, at this moment two generations of lead-free 

CBC ammunitions are now available. The first generation, mostly used in 

Europe, is almost exclusively made of strontium, together with smaller amounts 

of sodium, potassium and iron. The morphology is mostly consistent with GSR 

27

(spherical particles). The second generation, mostly used in the USA, contains 

no Strontium, only aluminium, silicon and potassium, and its morphology is not 

consistent with that of typical GSR particles – most particles being irregularly 

shaped without any particular morphology. This is in contradiction with other 

lead-free potassium-based primers from e.g. Winchester and Remington, which 

do produce spherical particles. 

Other occupational and environmental particles show the same elements and 

morphologies as these types of primers, and can be detected on e.g. automobile 

electricians, motor repair automechanics and petrol station attendants. Finally, 

also problems occur for the shooting distance determination, as the regular 

chemographic tests using rhodizonate do not react with the compounds in the 

primer or, as is the case for the modified Griess test, will also react with other 

nitro-compounds and hence can not be considered conclusive. 

3.1.2 Trends and comparisons 

Molina et al. (116) conducted an investigation of GSR testing results in suicide 

cases with analysis by Inductively Coupled Plasma-Atomic Emission 

Spectrometry (ICP-AES) and SEM/EDX. In this follow-up article on SEM/EDX 

analysis of suicide victims (117), in which a total of 116 cases of undisputed 

suicide by firearm were analysed, bulk analyses on 102 cases show a 

comparable result: only 50% of the samples tests positive. The authors conclude 

therefore that GSR analysis (by SEM/EDX or ICP-AES) can not be used to 

determine whether a person has shot a firearm, since a negative result does not 

signify that the person was not in contact with a shooting incident. Furthermore, 

GSR can not be used to distinguish whether a gunshot wound is self-inflicted or 

not. A positive result can be helpful, however, especially if used in concert with 

other information of the individual case. As a final observation, there is no 

significant difference between the performance of the SEM/EDX and bulk 

analysis techniques. Both tests have their benefits and deficiencies. 

3.1.3 Non-GSR sources of GSR-like particles 

Since a number of years concern has existed about GSR-like particles originating 

from a non-ballistic origin, which could lead to false-positive interpretation of the 

analysis results. A number of publications have already treated particles 

produced by car breaking systems and after the use of fireworks. These particles 

do not originate from the use of primers. Two publications, however, treated the 

characterisation of particles which arise after the non-ballistic use of primer 

compounds. 

Kinder and Provost (118) discuss a case where in a homicide, the suspect's 

defence was built around the fact that he has contact with Hilti primer-based nailguns. 

Although this type of equipment indeed uses a form of cartridges to drive 

the nail, the primer from a nail-gun cartridge was found to contain only lead and 

28

arium - no antimony. This precludes true three-component characteristic 

particles from originating from the small-calibre cartridges used in this type of Hilti 

nail-guns. 

Berk (119) addresses yet another non-ballistic source of GSR-like particles: car 

airbag inflation systems. Airbags may use primer materials in the deployment 

system, which may give rise to characteristic three-component GSR particles 

being emitted when the bag is ruptured. However, because materials are used in 

the airbag mechanisms that are normally not found in a firearm, the "GSR" 

particles will most of the time contain from trace amounts to large quantities of 

elements normally not encountered in true GSR particles. Moreover, because of 

the other materials present in an airbag, the overall population of particles on the 

samples will to a large extent be composed of GSR-inconsistent particles, giving 

away the non-ballistic origin of the particle deposits. The author concludes that 

the peculiar composition of these particles, both individually and as a population, 

ensures that it is normally possible to distinguish this population from true GSR - 

especially if the analyst is made aware that airbag material may have been 

involved in the case. 

3.1.4 Interpretation of analysis results and the application of Bayesian principles 

In a number of forensic disciplines, like DNA, fibres and mark-evidence, the 

application of Bayes’ Law for interpreting the analysis results in the context of the 

case at hand is already well established. Now, also in the GSR field voices are 

speaking up to implement these principles to give an added value to the purely 

analytical report that is often emitted after the analysis of GSR samples is 

completed. Biedermann et al. (120) offer a rigorous discussion of the principles 

involved and their implementation in the GSR field, based on statistical models 

developed by Cardinetti et al. (121). They also show how Bayesian Networks, a 

mathematical modelling technique originating from artificial intelligence research, 

may be used to model the several factors involved in the calculation of the 

likelihood ratio. The authors conclude that the Bayesian Law and Bayesian 

Networks serve as powerful tools to put the GSR analysis results in the 

perspective of the case context. The problem remains, however, that the 

numerical values to be assigned to the different parameters in the model need to 

be estimated and extracted from experimental data – an area of research that is 

to be further developed before these models can be reliably put to practical use. 

In this context a number of publications have appeared that further this part of 

development. 

Brożek-Mucha compared the difference in composition and morphology between 

airborne GSR and particles inside the cartridge case (122). In this study the 

composition and morphology of GSR particles originating from four brands of 

ammunition was compared. They represent three types of primer mixtures 

containing either mercury fulminate, lead styphnate or azide and an all-organic 

detonator. It is observed that GSR particles formed inside the case have not only 

29

a different shape, but may also show a different composition from the particles 

which were air-borne. Primers containing low-melting point metals like tin may 

produce particles which solidify after hitting the substrate (the surface of the 

cartridge case), although sometimes tin is present in the air-borne particles, but 

absent in the case particles. This suggests that tin in these particles originates 

not from the primer itself but from the bullet jacket. Thus, in the interpretation of 

the GSR analysis results not only the primer composition needs to be taken into 

account, but also the materials used in the manufacture of the bullet and jacket 

and the gun itself play an important role. Evidently, linking the composition of 

airborne GSR to the GSR in the cartridge case sometimes requires that 

laboratory experiments be performed under controlled conditions. This can of 

course only be done if the ballistic elements are present. 

In a second study, she investigated whether the GSR particle composition varies 

in different locations around the firearm/shooter (123). In this study, a Luger 9 

mm ammunition was fired from a pistol and samples collected from the shooter 

(hands, sleeves, front and back of the upper clothing) and the target at 50 cm. In 

a separate experiment, particles were collected from cotton targets, placed at five 

different distances up to one meter from the shooter. Trying to model the 

dispersion behaviour of the particles both in the shooting direction and in the 

opposite direction, links to the behaviour of the particles using the physical 

properties of dimension and density of the several types of particles were made. 

The author concludes that it is possible to observe differences in the composition 

of particles found on the hands of the shooter and on his clothing: the first 

originate from the ejection port of the gun, while the latter are partly coming from 

back-draft turbulence as the gaseous stream that follows the bullet through the 

barrel emerges from the muzzle and is distributed in all directions by the 

turbulent expansion in the air. The particles on the clothing of the shooter would 

therefore resemble more the particles found on a nearby target and contain more 

material coming from the bullet base, while the case/ejection port/hand samples 

would be constituted solely of material coming from the primer. 

Finally, Rijnders et al. (124) performed a comparable study using four types of 

ammunition (two classic, one titanium-zinc and one titanium-zinc doped with 

gadolinium) of the same calibre (9 mm) and one pistol. Samples were acquired 

from seven positions in and around the firearm, simulating both the shooter 

(clothing and hands), the target, and the gun/case internals. The particles were 

analysed by SEM/EDX and classified into 16 particle classes, specially designed 

for these experiments. Next, a statistical approach was used to calculate 

Pearson correlation coefficients. Using these statistics, a number of conclusions 

can be drawn: it is possible to distinguish between different ammunition types 

(from the set of chosen ammunition), internal samples from barrel and chamber 

correlate badly with external samples (hands, target, case) – an explanation for 

this may be found in the memory effect of the gun, while a good correlation can 

be seen in the samples of the shooters’ hand and victim. Also, according to this 

study, the results from the case samples correlated badly with the other samples, 

30

indicating that care must be taken when comparing samples taken from the 

suspects or victims and cartridge cases. This, of course is true on a purely 

quantitative level – it is clear that the elemental composition of the particles is still 

in agreement. 

3.1.5 Quality 

Due to international requirements (e.g. ENFSI) or national legislations, Quality 

assurance is more and more implemented in GSR laboratories, with a crucial 

step consisting of the accreditation of the analysis method following the ISO 

17025 norm. In reference (125) are discussed the type of documents that have to 

be prepared for this accreditation. Repeatability, reproducibility, robustness and 

interlaboratory tests are illustrated using an example of validation framework. 

Some Quality assurance considerations are also discussed when a new 

SEM/EDX equipment has to be acquired; on-site demonstration tests and 

selection criteria are proposed. 

Besides instrumental validation, other topics have to be examined for complete 

validation of GSR identification using SEM/EDX. Kopina (126) present a 

validation study including i) the characterization of GSR (composition and 

morphology) coming from 83 different ammunitions, ii) the effectiveness of 

sampling using different types of adhesives, and iii) the influence of time and 

activity after shooting. As a result of this validation, the load factor (amount of 

consistent particles per square millimetre of analysed sample) is introduced with 

a discriminating threshold value in order to distinguish background from potential 

discharge samples. Four classes are defined for each hand sample: unique and 

consistent, consistent, hand background, and contaminated (using a control 

sample). 

Proficiency tests are conducted every 2-3 years. They are organised by the 

ENFSI Expert Working Group "Firearms" and consist of the detection by 

SEM/EDX of 100 three-element particles (lead, barium and antimony) distributed 

in four size classes (0.5 µm, 0.8 µm, 1.2 µm and 2.4 µm). Two proficiency tests 

were conducted during the period of interest (GSR2008 and GSR2010). The 

results have been (GSR2008) or will be (GSR2010) discussed during dedicated 

workshops. Moreover, the results of the GSR2005 proficiency test were 

published (127). Compared to the previous test (GSR2003 (128)), more 

laboratories participated and about one fourth of them were non-European 

laboratories (mostly USA). The comparison of the z-scores of the two tests 

revealed an improvement of the detection capability of the laboratories. Moreover 

the results are on average better for the laboratories that took part in the two 

series, leading to the conclusion that a continuous participation in the proficiency 

test program contributes to an improvement in the individual performances. 

31

3.2 Estimation of shooting distance and bullet hole characterization 

3.2.1 General considerations 

The greatest part of GSR produced by a shooting is projected on the target 

(object or victim) if this target is close enough to the shooter. The diameter and 

the density of the GSR particle deposition pattern will help to determine the firing 

distance. Moreover, the rotating bullet will usually produce a wipe ring around the 

entrance hole. The presence or not of a wipe ring will help to state on the nature 

of the bullet hole (entrance or exit). 

When the nature of a wound is questionable (entrance or exit wound), some 

research groups recommend to look at the presence of textile fibres (coming 

from the victim’s clothing) since in case of an exit wound, textile fibres should be 

absent. However Vennemann et al. (129) demonstrated that the presence of 

textile fibres is not a suitable criterion to determine the nature of a wound since 

the presence of textile fibres both from the entrance and exit holes could be 

stated; moreover the density of the fibres along the bullet path seems to be 

dependent of the length of this path and of the size of the temporary cavity. 

3.2.2 Colour tests 

Colour tests are usually needed to determine the firing distance and the nature of 

bullet holes since the GSR pattern can be invisible due to lack of contrast 

(especially for dark fabrics). Distance determination using colour tests are 

essentially based on visual examination with the use of test firings at known 

distance for comparison. The use of the same weapon and the same ammunition 

is crucial since it will strongly affect the GSR pattern. This has already been 

pointed out in many references. Another illustration of the influence of weapon 

and ammunition is brought in reference (103). 

The colour test of choice for the estimation of shooting distance and bullet hole 

characterization is still the sodium rhodizonate test, specially sensitive to lead. 

For instance a positive test with sodium rhodizonate is indicative of an entrance 

hole. Bailey et al. (130) evaluated the limit of detectability, determined to be 

approximately 160 ng Pb/cm². The test was positive for 35 different .22 calibre 

ammunition with a shooting distance of 7.6 m, indicating a strong robustness of 

the sodium rhodizonate test. In another study, Bailey et al. (131) tested 70 holes 

due to air gun pellets, presumably less sensitive to the sodium rhodizonate test 

since the wipe rings do not contain cartridge components such as primer 

residues. About 60% of the pellet holes yielded a positive reaction for lead. 

Haag (132) performed a study on ricochet marks. SEM/EDX was used in 

conjunction with sodium rhodizonate and dithiooxamide (a colour test used for 

the detection of copper). The use of SEM/EDX enhanced the initial information 

obtained from the colour test by giving additional information that can help in 

32

shooting incident reconstruction. Using a similar strategy in the particular case of 

a victim shot in the head, the examination of hairs using SEM/EDX can give 

information about the firing distance since at close distance GSR particles were 

found in the inner surfaces of the cuticle (133). 

Lead Check® swabs – a commercial test based on the sodium rhodizonate test 

and developed at first for in field detection of high concentrations of lead in paint 

and building materials – have been tested for crime scene application (134). The 

swabs were applied on different materials (for effectiveness) and with different 

metals (for selectiveness). It was shown that the test performed equally well, 

compared to the traditional sodium rhodizonate test. In the same manner two 

other commercial tests for lead and copper have been tested: Plumbtesmo® test 

(designed for lead deposits in motor vehicles tailpipes) and Cuprotesmo® test 

(designed for detection of copper on surfaces) (135). It was shown that i) the 

Plumbtesmo® test performed as well as or even better than the sodium 

rhodizonate test, and ii) the Cupresmo® test and dithiooxamide had comparable 

performances. The facility of use makes these tests well adapted for in field 

analysis. 

The examination of histological sections can also give an idea about the firing 

distance: In reference (136), the histological sections were stained using sodium 

rhodizonate in order to count and measure the GSR particles in the epidermal 

and subepidermal layers. A relation between firing distance and the size of 

particles could be pointed out. 

Some procedures using the sodium rhodizonate test recommend the application 

of a 15% acetic solution onto a piece of filter paper that is then pressed against 

the area of interest (transfer method). The transport of an acetic solution can be 

problematic (e.g. travel by air) when the tests have to be performed in the field. 

Lau (137) suggest the replacement of 15% acetic acid by commercial vinegar 

available from any supermarket. However if lead around the bullet hole is 

correctly detected using this reagent, the lead cloud (for distance determination) 

around the hole may not be detected. 

The Griess test is a colour test sensitive to nitrites. It is routinely used in many 

forensic laboratories but is not as developed as the sodium rhodizonate test 

since it suffers from some limitations. Lau (138) tested different formulations and 

replaced successfully the desensitized photographic paper usually used for 

transfer medium by blotter paper. 

Colour tests usually necessitate transfer techniques rather than direct pattern 

development since the contrast could not be high enough for dark fabrics. 

Consequently the lack of precision of the colour tests can be due to the loss of 

GSR during the transfer, causing an overestimation of the distance of firing when 

comparing with test firings. In order to minimize this issue, De Forest et al. (139) 

propose a direct detection of GSR based on the property of lead (and antimony) 

33

fluorescence in an acidic media at low temperature. In contrast, Vinokurov et al. 

(140) examined the possibility of a contamination of the victim’s clothing, 

inducing a negative bias, for instance when the victim falls on the contaminated 

surface between her and the shooter. However it was found that the transfer is 

quite low and misinterpretation can thus only occur for relatively long distances (> 

1 m) when few GSR are visualized in the vicinity of the hole. 

3.2.3 Development of analytical techniques 

Beside visual examination intrinsically subject to lack of precision, analytical 

methods are developed to improve the accuracy of the results. Cagliano-Candela 

et al. (141) used atomic absorption spectroscopy to quantify lead concentration 

around bullet holes. A linear relationship was found between the firing distance 

and the logarithm of lead amount in the outer ring around the hole. 

In a similar approach, Sharma and Lahiri (142) used FTIR microscopy for 

distance determination (7-30 cm) based on the concentration of nitroglycerine. 

Reid et al. (143) took a look at the sampling dilemma: whether to use swabs or 

stubs for GSR analysis by SEM/EDX. In a statistical analysis of the results of 

using stubs and alcohol-imbibed swabs to collect GSR from hands, authors show 

that the adhesive stub method is definitely better suited for collection of this 

particulate material. 

Souza Sarkis J. et al. (144) Further developed the use of sector field ICP-MS to 

analyse lead, barium and antimony levels on test shooters and controls in a 

study carried out using .38 revolver, and 9mm and .40 calibre pistols. Realising 

that absolute quantification and recovery studies are of no use, the authors use 

ternary plots to show a presence of the trace elements that can lead to 

identification of GSR on shooters’ hands. 

Finally, Rosenberg et al. (145) determined the detection time frame of inorganic 

GSR by a preliminary field test using LASER-Induced Breakdown Spectroscopy 

(LIBS). Although LIBS is a bulk analysis technique for inorganic materials, it can 

be used, according to the authors, as a presumptive and inexpensive field test, 

after which the SEM/EDX technique would complement it with its high specificity 

for characteristic GSR particles. The authors claim that up to 5 days after firing, a 

significant barium signal can still be measured on the hands of test shooters who 

had been living normal lives, including hand washing and other day to day 

activities. 

3.3 Organic GSR 

Next to the inorganic components of GSR, also the organic part of the residues is 

more and more being studied for use in routine investigations. Where in the past 

predominantly chromatographic techniques were developed (both in liquid and 

34

gaseous state), we see now a diversification into other, ultra-modern, techniques 

such as LASER Spectroscopy and DESI probes. 

3.3.1 Screening of gun propellant powder components 

NIST (146) has produced a Standard Reference Material with known 

concentrations of nitroglycerin, (N-nitroso)diphenylamine and ethylcentralite. This 

material can be used to validate analysis methods in an accreditation effort of 

forensic laboratories, or as a proficiency standard. NIST is currently developing 

two more standards aimed at the determination of trace amounts of high 

explosives. One is developed for calibration of detection equipment as used in 

airport security, and the other for measurements associated with remediation of 

contaminated soil. 

Haag et al. (147) have investigated the new Blackhorn 209 propellant, a novel 

formulation that is a substitute for black powder in muzzle-loading firearms. It 

provides bullet speeds slightly higher than the equivalent modern black powder 

formulations, but leaves mildly corrosive residues in the bore of the firearm and in 

the cartridge cases. This new powder is unlikely to turn up in many cases due to 

high cost and narrow application, but may turn up in cases involving gun bursts 

and pipe bombs 

3.3.2 Instrumental methods 

A number of techniques have been used to detect organic residues in different 

applications and varying substrates. Among them, Solid Phase Micro-Extraction 

(SPME) is used in combination with GC or Ion Mobility Spectrometry (IMS). 

Weyermann et al. (148) used GC-MS analysis after SPME sampling of fired 9 

mm cartridge cases. They were able to identify thirty two organic volatile 

compounds, some of which could be traced until up to 32 hours after the shot. 

The problem is that, as work in this area is very original, the exact origin of most 

of these compounds is still unknown. The study shows further that the largest 

problem involved with the use of SPME is the reproducibility of the 

measurements of nanogram-range quantities of material. Joshi et al. (149), on 

the other hand, successfully used SPME and Ion Mobility Spectrometry for the 

detection of odour signatures of smokeless powders. Besides being an 

instrumental alternative to canines for the detection of compounds used in homemade 

explosive devices, their method detects also components for which the 

dogs are not useful. For instance, 2-ethyl hexanol and 2,4-DNT were detected by 

canines, while diphenyl amine (DPA) and ethylcentralite (EC) were not of interest 

to them. As IMS instruments are readily available, e.g.. in airports, where they 

are used for detection of illicit substances, these results therefore complement 

the canine detection by instrumental techniques. 

All these techniques have in common that the analytes are being sampled and 

concentrated on a carrier substrate prior to analysis. On the other hand, 

35

Scherperel et al. (150) show a method for extraction and characterisation of 

smokeless powder additives (methylcentralite (MC), EC and DPA) using tandem 

mass spectrometry (nESI-MS), without the use of chromatographic separation 

methods. From seven types of gun powder in unburned form, only two could not 

be distinguished based on physical appearance of the unburned grain, solubility 

in methanol and the nESI-MS technique. Samples from the burned powder were 

used to successfully identify low levels of EC using Collision-Induced 

Dissociation (CID)-MS/MS. The authors conclude that nESI-MS has great 

potential for application in GSR analysis. 

Zhao et al. (151) demonstrate the applicability of Desorption ElectroSpray 

Ionisation (DESI)-MS/MS spectrometry, applied directly on the surface to be 

sampled, for the detection of stabilizers (MC, EC, DPA). The DESI sampler can 

even be applied directly on the human skin and the analysis carried out using a 

portable MS/MS - making this potentially a field-operation technique. 

Contamination of the surface by milk, soft drinks and dust does not influence the 

measurements. Other surfaces were also tested, such as glass, rubber and 

leather gloves, towel, cotton and hair. As far as sensitivity and persistence are 

concerned, MC could still reliably be detected for up to 12 h after the shooting, 

and it could still be detected after 6 hand washings. 

3.4 Book review 

Adlard (152) in a review of a book by Wallace “Chemical Analysis of Firearms, 

Ammunition and Gunshot Residue”, describes the collective knowledge and 

experience gathered by the scientists at the Northern Ireland Forensic Science 

laboratory. The book compiles the results obtained using the diverse instruments 

available for the analysis of GSR and, to a lesser extent, explosives. 

36

4. References 

1. National Research Council (U.S.). Committee on Identifying the Needs of 

the Forensic Sciences Community; Committee on Applied and Theoretical 

Statistics. Strengthening Forensic Science in the United States: A Path 

Forward. The National Academies Press; 2009. An executive summary can 

be downloaded free of charge: 

http://ag.ca.gov/meetings/tf/pdf/2009_NAS_report.pdf 

2. AFTE Committee for the Advancement of the Science of Firearm & 

Toolmark Identification. The Response of the Association of Firearm and 

Tool Mark Examiners to the February 2009 National Academy of Science 

Report “Strengthening Forensic Science in the United States: A Path 

Forward.” June 22, 2009. AFTE Journal 2009; 41 (3):204-208 


Toolmark Identification. AFTE Response to the NACDL Task Force on the 

Future of Forensic Science. AFTE Journal 2010; 42 (2):102-107 

4. Buckleton J, Triggs C, Taroni F, Champod C, Wevers G. Experimental 

Design for Acquiring Relevant Data to Address the Issue of Comparing 

Consecutively Manufactured Tools and Firearms. Science & Justice 2008; 

48 (4):178-181 

5. Lyons DJ. The Identification of Consecutively Manufactured Extractors. 

AFTE Journal 2009; 41 (3):246-256 

6. Hamby J, Thorpe J. The Examination, Evaluation and Identification of 9mm 

Cartridge Cases Fired from 617 Different GLOCK Model 17 & 19 

Semiautomatic Pistols. AFTE Journal 2009; 41 (4):310-324 

7. Uchiyama T. Toolmark Reproducibility on Fired Bullets and Expended 

Cartridge Cases. AFTE Journal 2008; 40 (1):3-46 

8. Gouwe J, Hamby JE, Norris SA. Comparison of 10,000 Consecutively Fired 

Cartridge Cases from a Model 22 Glock .40 S&W Caliber Semiautomatic 

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9. Hamby JE, Brundage DJ, Thorpe JW. The Identification of Bullets Fired 

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of Forensic Sciences 2008; 53(4):807-811 

152. Adlard E. Review of “Chemical Analysis of Firearms, Ammunition and 

Gunshot Residue”. Chromatographia 2008; 68 (9/10):882 

47

Examination of Marks 


Nadav Levin, MSc 

Head, Toolmarks and Materials Laboratory 

Division of Identification and Forensic Science (DIFS) 

Israel National Police Headquarters 

Jerusalem 91906, Israel 

E-mail: simanim@police.gov.il (Lab), 

Nadav.levin@gmsil.com (Privet) 

48

Table of Contents 


2  FOOTWEAR AND TIRE‐TREAD IMPRESSIONS  51  

2.1  Detection and Recording  52  

2.1.1  Photography and Image Processing  52  

2.1.2  Lifting and Casting  54  

2.1.3  Shoeprints in Blood  55  

2.1.4  Chemical Enhancement  57  

2.1.6  Shoeprints and Tire‐tracks in Snow  57  

2.1.6  Imprints on Miscellaneous Surfaces  58  

2.2  Manufacturing Processes and Outsoles Design  58  

2.3  Tire Tracks  59  

2.4  Test Impressions  59  

2.5  The Evidential Value of Shoeprints Examination  60  

2.6  Databases, Reference Collections and Automated Classification  61  

2.7  Miscellaneous  63  

3.  TOOLMARKS  63  

3.1  Casting and Reproduction Methods  64  

3.2  Observation and Imaging Methods  65  

3.3  Marks Produces by Various Types of Tools  66  

3.4  Examination of ConsecutivelyManufactured Tools  68  

3.5  The Examination of Stabbing and Cutting Marks  69  

3.6  Evidential Value of Toolmark Examination  70  

3.7  Miscellaneous Issues  72  

4.  PHYSICAL MATCH  73  

5.  RESTORATION OF OBLITERATED MARKS  75  

5.1  Steel surfaces  75  

5.2  Aluminium alloy surfaces  76  

5.3  Laser engraved marks  77  

5.4  XRay Radiography  77  

49

6.  REFERENCES  78  

50


The examinations of contact marks and related topics covered in this review are 

among some of the core issues of forensic science. Although these types of 

evidence may sometimes have the highest evidential values in definitely linking 

suspects to scenes of crime, the techniques used are usually straightforward and 

simple. This review covers advances in scientific methods applied to the forensic 

examination of various marks, since the Interpol 15 th International Forensic 

Science Symposium (IFSS) in October 2007 (1). 

This paper is based mainly on a literature review, derived from the UK Forensic 

Science Service (FSS) FORS database (Forensic Bibliography Database), which 

covers articles published in the principle forensic science journals and other 

relevant sources over the review period (2). This has been supplemented by a 

search of the Internet for articles related to the forensic examination of marks, 

using, for instance, the Google Scholar search engine (3). Manuals and standard 

operating procedures of various forensic science laboratories, many of which are 

relevant to this Review, may also be found on the Web, for instance – the Virginia 

Department of Forensic Science (VA-DFS) ones (4). Since one of the purposes of 

this Review is to provide a wide updated background for practitioners in these 

fields, some publicly-available Internet references are also listed below. 

2 Footwear and Tire-Tread Impressions 

Footwear impressions may be considered, apparently, as one of the most 

common types of evidence, and are found, virtually, in every scene of crime. As a 

significant form of physical evidence, impressions left behind at the crime scene 

may provide valuable information on where the crime occurred and the direction 

the suspect travelled while committing the crime. This information may place the 

suspect at the crime scene or eliminate him as having been there. A general 

review of this field was published by Smith (5), covering both the class and 

accidental characteristics found in shoeprints, and the evidential value of the 

examinations’ outcome. Although general in nature, this article provides a good 

starting point for novice in this area. 

The Scientific Working Group on Shoeprint and Tire Tread Evidence 

(SWGTREAD) continues its effort for setting professional guidelines for the 

collection, preservation and examination of footwear and tire tread impression 

evidence (6). The Group’s new Internet web-page provides a vast amount of 

information, including some recently-approved guides: 

• Guide for Casework Documentation (September 2008), 

• Guide for the Chemical Enhancement of Bloody Footwear and Tire 

Impression Evidence (September 2008), 

• Terminology Used for Forensic Footwear and Tire Impression Evidence 

(March 2009). 

These guides, along with the previously-published ones, may be downloaded freeof-charge 

from the SWGTREAD web-page. In addition, a “Daubert Resource Kit”, 

51

dealing with the presentation of shoeprint evidence in court, may also be provided 

by this Group. 

Useful guides, regarding various aspects of the photography of footwear marks, 

are also found at the Scientific Working Group on Imaging Technology (SWGIT) 

web-page (7). Among these guides are: 

• Field Photography Equipment and Supporting Infrastructure (2009, 

updated version), 

• Guidelines for Image Processing (2010, updated version), 

• General Guidelines for Photographing Tire Impressions (2010, updated 

version), 

• General Guidelines for Photographing Footwear Impressions (2010, 

updated version). 

The European Network of Forensic Science Institutes (ENFSI) Expert Working 

Group Marks (EWGM), established in 1995, also runs an active web-page (8). 

One of the remarkable features of this web-page is its “Wanted Page”, enabling 

the exchange of information regarding unknown shoeprints. Since the beginning of 

2008, more than 150 queries were submitted by shoeprint examiners from all over 

the globe, many of which were successfully answered by other peers. In addition, 

this Group’s newsletter, “Information Bulletin for Shoeprint/Toolmark Examiners” 

(IBSTE) is also posted on this web-page. 

The Virginia Department of Forensic Science (VA-DFS) has prepared a 

procedures manual of various laboratory methods for footwear and tire tread 

impressions, as well as a training manual for experts in this field (4). These 

documents, as other VA-DFS manuals, are available on the Internet. 

The Home Office Scientific Development Branch (HOSDB, UK) Fingerprint and 

Footwear Forensics (FFF) group produced special edition newsletters (9, 10), 

dedicated to the recovery and imaging of footwear marks. These newsletters 

comprise comprehensive and practical guidelines for many aspects of lifting 

techniques and chemical enhancement methods for shoeprints found on various 

surfaces. 

2.1 Detection and Recording 

2.1.1 Photography and Image Processing 

Footwear impressions can be photographed (on film or digitally) or scanned. The 

standard methods for shoeprint photography are covered in numerous articles, 

including the SWGTREAD and the SWGIT guides (6, 7). 

Blitzer and his colleagues (11) conducted a comprehensive study regarding the 

effects of the photographic technology on the examination quality of footwear 

impressions. The purpose of their study was to determine how footwear 

impression examiners respond to different photographic methods employed. The 

researchers used both resolution standard targets and inked shoeprints for the 

52

purpose of this study. Examiners having at least some experience with footwear 

impressions analysis were involved. Images were taken using a high quality (14 

megapixels) digital camera and a medium quality (6 megapixels) one, a 35mm film 

camera and a 120 format film camera. Digital images were taken in the “zero 

compression” JPEG format. The conclusions of this study were as follows: 

• Both digital cameras of 6- or 14- megapixels are satisfactory substitutes 

for the 35mm film camera for most regular shoeprint cases, 

• In those special cases where higher resolution is required, a 120 format 

film camera is recommended, 

• There is not yet a simple and suitable digital solution for life-sized prints. 

Chung (12) describes a method of photographic enhancement of difficult-tocapture 

two-dimensional (2D) prints by the combined effect of overhead soft box 

lighting and the Tilt-Shift lens (perspective control lens) presented earlier by this 

author (13). This combined method was tested on different types of impressions, 

on different substrates, and was found to be superior over traditional photographic 

methods. 

Brown and et al (14) examined the forensic application of high dynamic-range 

(HDR) photography. These authors used Photoshop CS4 and Photomatrix Pro 3 

software for combining several (3 – 5) images taken in different exposures, 

resulting in high-quality 32-bit images. One of the examples presented in this 

article was of a shoeprint. Combining multiple exposures of a 3-dimentional (3D) 

shoeprint into a single HDR image allows observing a detailed print. This method 

is also useful for capturing prints on a multicolor background (like wallpaper, 

magazine cover, etc.) 

The 2010 HOSDB newsletter mentioned above (10) contains, among other useful 

and practical information, a best practice guide for the imaging of footwear marks, 

including recommendations for optimum imaging techniques for various types of 

marks and surfaces. 

Buck and her colleagues (15) used a high resolution 3D optical scanner for 

recording footwear impressions and tire tracks in snow. Their results proved that 

non-destructive 3D optical surface scanning is a suitable method for the 

documentation of impressions in snow. The 3D models of the scanned 

impressions in snow displayed a high accuracy including all the fine details. 

Casting footwear impressions in snow has always been a difficult assignment for 

forensic identification specialists, and the 3D optical surface scanning is 

apparently an accurate and efficient new method for documenting impressions in 

snow without contact. Effective results can be achieved even in different kinds of 

snow and under various meteorological conditions. The method is also suitable for 

impressions in soil, sand or other materials 

Polynomial Texture Maps (PTMs) are a simple representation for images of 

functions instead of just images of color values. In a conventional image, each 

pixel contains static red, green or blue values. In a PTM, each pixel contains a 

simple function that specifies the red, green or blue values of that pixel as a 

53

function of two independent parameters, specifying the direction of a point light 

source. PTMs are typically produced with a digital camera by photographing an 

object multiple times with lighting direction varying between images. Even a lowend 

digital camera provides enough resolution to produce good PTMs, and almost 

any light source can be used, such as a light bulb, LED or a flash. Hamiel and 

Yoshida (16) applied this imaging technique for shoeprints and other impression 

evidence, including the use of a portable unit for field studies. The PTM images 

were compared to conventional sidelight and casting techniques. The application 

of this technology could be more cost-effective than conventional methods and 

provide higher-quality data. Results of the evaluation reveal that PTM technology 

can successfully be used in the forensic field and has the potential to produce 

better resolved images for the comparison of known shoe soles or tire treads to 

crime scene impressions. Specific results indicated that PTM images and 

enhancements improved the visibility of detail in some of the impressions under 

analysis when compared to traditional photography techniques, including 

improvement of the visualization of texture within a shoe or tire impression. PTM 

technology thus gives the examiner the best opportunity for visualizing unique 

characteristics in impression evidence. PTM technology also has the advantage of 

being cheaper to operate than traditional sidelight and casting techniques. 

In a recent report by Prokoski (17), 2D infrared (IR) imaging was demonstrated to 

produce images of footwear impressions under dim light and under total darkness 

conditions. It was also shown to produce detailed images of athletic shoe sole 

patterns without controlled lighting. 3D IR imaging was also demonstrated to 

produce dimensionally accurate 3D digital models of footwear and footwear 

impressions simply and fast. The primary advantage of IR imaging over visible 

light imaging for footwear evidence is that it produces more consistent feature 

details under conditions of uncontrolled lighting. Feature contrast in IR images can 

generally be enhanced by air flow over the features, which differentially heats or 

cools portions of the features. 

Since digital photography is becoming more and more common for capturing 

shoeprint images, readily-available image enhancement enables the shoeprint 

expert observing details on the shoe-mark that were initially barely visible. Wiesner 

and her colleagues (18) presented two Collaborative Testing Services (CTS) 

proficiency tests (04-533 and 07-533), where challenging shoeprints were 

examined. The prints were made on surfaces with noisy backgrounds that 

interfered with the prints. The image processing method presented in this paper, 

namely color channel separation and image enhancement, reduced the 

background significantly, and improved the visibility of the shoeprints. 

2.1.2 Lifting and Casting 

Following photography or scanning, shoeprints found on various surfaces are 

usually lifted or cast, according to their nature. Recently published SWGTREAD 

guides cover this area (6). Footwear impression examiners usually use 

transparency lifts to compare size, design and accidental characteristics between 

known shoes and questioned impressions. In certain cases, the examiner may 

have to separate the lifting film from the cover sheet. In a case reported by Adair 

54

(19), which involved “Lightning Lift” transparency lifts (by Lightning Powder 

Company, USA), such a separation resulted in the apparent reversal of the 

orientation of certain elements of the outsole, indicating that the adhesive and 

impression had been transferred from the thinner lifting sheet to the thicker cover 

sheet. Tests with other “Lightning Lift” transparencies from previous casework 

revealed the same phenomenon of transference of the image and adhesive. 

Awareness of this phenomenon, and its implications for the interpretation of 

footwear impression evidence collected using such transparencies, are advised. 

Examiners may sometimes be faced with footwear marks found on various types 

of substrates. A murder case where a partial print was found on a coffee 

polystyrene cup was presented by Bekiempis (20). Numerous methods for 

recovering the print from the polystyrene surface were tested. The most successful 

one was using a dental stone cast, while Forensic Sil silicone rubber (Loci 

Forensic Products, The Netherlands) gave also very good results. It is highly 

recommended by this author always to take good quality photographs of the prints 

prior to the application of casting or enhancing methods. 

Isomark (UK) offers a novel product, Foot-Print, for taking 3D footwear 

impressions – silicone rubber in a dispensing gun (21). According to the 

manufacturer, typical curing times for the Foot-Print are between 3 and 10 

minutes. This product is also suitable for shoeprints in snow, where the curing time 

may reach a maximum of 20 minutes in freezing temperatures. 

A new device for scanning gelatine lifters, GLScan, was introduced by BVDA (The 

Netherlands) (22). The device, based on a line-scan camera, has a resolution of 

1046dpi, higher than any available digital camera, and it also offers evenly 

illuminated images, image enhancement software, ease of operation and high 

throughput. According to this manufacturer, scanning a gelatine lifter 18x36cm in 

size takes about 2.5 minutes. 

2.1.3 Shoeprints in Blood 

The recovery of footwear impressions in blood is similar, in most cases, to the 

recovery of fingerprints in blood, and indeed many of the relevant references cover 

both these areas of interest. This section will focus mainly on those articles 

referring to shoeprints. 

As mentioned above, SWGTREAD has published a comprehensive guide for the 

chemical enhancement of bloody footwear and tire impression evidence in the field 

and in the laboratory (6), including a list of appropriate reagents and instructions 

on their application. 

Cullen et al (23) have recently published an extensive article, describing the 

results of a controlled experiment, where about 50 identifiable shoeprints in blood, 

deposited on various substrates, were buried in soil for up to four-week time. 

About 60% of the prints were visible when excavated, although poor recovery 

rates and loss of impressions were observed on substrates buried for more than 

two weeks. Chemical enhancement methods were then applied to the prints, and 

55

more than 70% of the prints provided adequate visible impressions that were 

identifiable to the original ones. The most effective methods for the enhancement 

and retrieval of impressions were leucocrystal violet (LCV) and Bluestar. This 

study concluded that a significant amount of blood-contaminated footwear 

impressions can be recovered from buried substrates, provided that careful 

excavation methodology and suitable enhancement techniques are utilized. 

Methods for the chemical enhancement of footwear marks in blood are also 

discussed in the 2008 HOSDB newsletter mentioned above (9). This article 

recommends the application of protein stains (like Acid Violet 17) as a speculative 

search tool for bloody footwear marks on non-porous flooring surfaces, provided 

the conditions are suitable. 

Farrugia and his colleagues (24) are about to publish an article regarding the 

chemical enhancement of shoeprints in blood, recovered from fabric surfaces 

using alginate casting materials. Since bloody footwear impressions are found 

sometimes on colorful fabrics, the in-situ enhancement provides in such instances 

less-than-desired results. In order to overcome these limitations, this study 

examined the lifting of the marks by using alginate casting materials followed by 

chemical enhancement by the Acid Black 1 (Amido Black) and LCV reagents. 

These authors recommend the use of Amido Black for this purpose, due to the 

safety issues involved when using LCV. 

Footwear impressions in blood deteriorate over time, even when indoors or 

sheltered from adverse environmental conditions. Morgan-Smith et al (25) tested 

the influence of aging of shoeprints in blood on their enhancement using various 

methods. These researchers used three distinctly different shoe sole pattern styles 

for creating test impressions and added fine damage features to each sole in order 

to assist the evaluation of the results. Shoeprints were placed on several surfaces, 

including linoleum, varnish wood and paper, and aged for up to 16 weeks in 

various environments. The prints were then treated with six different chemical 

reagents for blood enhancement. Results showed that, of the methods compared, 

ninhydrin was the best reagent for treating aged impressions on paper substrates. 

However, on wooden and linoleum surfaces, amido black was the reagent of 

choice. This study also demonstrated that shoeprints in blood do deteriorate over 

time, especially outdoors, and that impressions on paper showed the least 

deterioration. 

Gorn et al (26) presented a case where footwear impressions in blood were found 

at a homicide scene involving a smoldering fire. This case initiated a study aimed 

at the specificity of LCV for distinguishing blood from other liquids that were found 

on the scene. As the results showed, LCV is highly specific to blood, and none of 

the other tested substances, apart from blood, gave positive results with it. It is 

important to note that the reaction of LCV with blood occurs within one minute. 

Gelatine lifters (gel lifters) are not considered, usually, as the preferred method of 

choice for lifting impressions in blood. Nevertheless, Svejderud and Lundqvist (27) 

presented a case where such lifters were successfully applied for collecting 

footprints from wooden floor tiles. The relevant footprint was first lifted using black 

56

gel lifter, followed by a white one. When the lifters where taken to the authors’ 

laboratory and photographed, the photographer noticed by coincidence that 

although the prints on the gel lifters were quite poor, there were distinct prints on 

the transparent protective plastic sheet of each of these lifters. Interestingly, the 

second (white) lifter showed a better print than the first (black) one, probably due 

to the removal of interfering background by the first. 

2.1.4 Chemical Enhancement 

The HOSDB 2008 newsletter mentioned above (9) contains also a summary of 

methods for enhancement of shoeprints made of various substrates. The methods 

tested in this study were those routinely used for latent fingerprints development. It 

is interesting to note that a powder suspension was very effective on most 

contaminants, and was the single most effective process on 50% of the tested 

contaminants. 

McNeil and Knaap (28) compared the performances of the bromophenol blue 

(BPB) indicator with those of potassium thiocyanate for the enhancement of dust 

footwear impressions. These authors found that BPB performed better than 

potassium thiocyanate on most shoeprints tested. 

2.1.6 Shoeprints and Tire-tracks in Snow 

Taking casts of shoeprints or tire-tracks in snow is more challenging than other 3D 

impressions, due the nature of this substrate. The commonly-used dental stone is 

not applicable for snow impressions due to its exothermic reaction while curing. 

Numerous methods were published over the years for such cases. One of these 

methods was to use molten pure sulphur, which is well studied and documented. 

There are, however, several limitations in using this method: It requires additional 

equipment (a stove and a melting pot) not usually carried as a part of an evidence 

collection kit, a respirator is needed due to the hazardous sulphur dioxide fumes 

produced, and the produced sulphur casts tend to be brittle. In order to overcome 

some of these limitations, Wolfe (29) studied the use of sulphur cement for casting 

snow impressions. Sulphur cement is a silica-filled modified sulphur mixture, which 

is melted, cooled and poured in the same manner as pure sulphur, but has higher 

strength and stability. Validation studies showed that sulphur cement could rapidly 

and reliably preserve snow impression evidence with detail comparable to that of 

pure sulphur or dental stone casts. Based on this study, sulphur cement proved to 

be safer and of higher strength for casting snow impressions, as a substitute for 

pure sulphur. 

In order to overcome the disadvantages of dental stone for casting impressions in 

snow, Adair and Shaw (30) proposed a dry-casting method. When applying this 

method, these authors used a commercial flour sifter to sift the dry dental stone 

powder onto the impression in fine layers, followed by cold water spraying (using a 

plastic spray bottle). The dry-casting method is remarkable for several reasons: 

First, the method uses materials commonly carried by crime scene investigators, 

namely dental stone and water. Second, flour sifters are inexpensive and simple to 

57

use. The technique is not adversely affected by cold temperatures and works well 

on a variety of snow pack conditions. More important, however, is the quality of the 

final cast when compared to the application of dental stone by pouring. 

Understanding the medium of snow and how its properties may influence the 

success of casting methods can assist the investigator in choosing techniques that 

offer the best chance for successfully casting track impressions. Additionally, snow 

conditions may change rapidly because of the influence of solar heat, wind, 

additional precipitation, contaminants, and combinations thereof. Adair and his 

colleagues (31) discussed the various snow types and present recommendations 

for the appropriate casting methods best suited for the characteristics of the 

snowpack. 

2.1.6 Imprints on Miscellaneous Surfaces 

Chochol and Swietek (32) present three murder cases of partial shoeprints 

detected on a human body. In two of the cases, the prints were photographed 

during the autopsies and compare to suspects’ shoes. In the third case, no 

suspect shoe was available, so only the brand of such a shoe had to be 

determined. These authors showed that it is sometimes possible to perform 

shoeprint analyses even on such difficult surfaces as human skin. However, it 

should be emphasised that most often partial prints are found, and only limited 

class characteristics can be identified. Such footwear impression may also be 

used for reconstructing the incident, placing the perpetrator in relation to the 

victim, and so on. 

An uncommon case of finding a break pedal pad impression, of a vehicle involved 

in a fatal accident, on the shoe outsole of one of the suspects, is reported by White 

(33). In the case reported, both of the occupants of a vehicle that was involved in a 

fatal collision denied being the driver. The author reported that impression found 

on the outsole of one of the suspects matched, by class characteristics, the 

pattern of the vehicle’s break pedal pad. Based on these findings, among others, 

that suspect was convicted. 

2.2 Manufacturing Processes and Outsoles Design 

As in most areas of forensic comparisons, understanding of manufacturing 

processes is essential for shoeprint examinations. Good sources of information for 

this aspect are the footwear industry newsletters, like the SATRA Bulletin. In the 

Review period, several articles have been published in this Bulletin regarding the 

manufacturing of moulded boots, of injection-moulded soles and of leather soles 

(34 - 36). Although the addressees of these articles are mainly members of the 

footwear industry, forensic scientists dealing with shoeprint comparison may gain 

great benefits from them as well. 

The individualization of footwear and tires rests mainly upon the knowledge that 

accidental marks formed during the use, or abuse, are unique. Thus, 

manufacturing defects, mistakenly identified as unique wear damage, may lead 

the examiner to the wrong conclusions. Adair (37) describes a case of new, 

58

unused, deck shoes of poor manufacturing quality presenting manufacturing 

defects resembling in shape wear accidental defects. This example supplements 

previously published articles calling for caution in this respect. Although such 

cases may be rare, inexperienced examiners should be aware of the potential for 

manufacturing defects, especially when examining inexpensive footwear. Here, 

again, knowledge of the manufacturing processes involved is essential. 

Knowledge regarding the thread design of footwear outsoles is also crucial for 

shoeprint identification. Adair (38) also reports a case where shoeprints in blood 

were found in an assault scene. The impressions were comprised primarily of a 

“double-helix” style pattern travelling lateral-medialy across the outsole. Some 

“arms” of the double-helix pattern were incomplete, which suggested that one arm 

might be slightly higher than the other. Initial search of outsole databases was 

negative. The outsole pattern was eventually identified as a Nike Jordan B’2rue 

brand shoe. The shoe pattern is remarkable in that one arm of the double-helix 

design is slightly elevated above the other. This design is relevant to footwear 

examiners since the impressions created by new shoes will appear quite different 

than impressions created by heavily worn shoes. In a new, relatively unworn state, 

the impressions created by this shoe will have only one arm of the double-helix 

present in 2D impression. As the shoe wears, the pattern will begin to resemble 

the double helix pattern. 

2.3 Tire Tracks 

Bodziak (39) published a comprehensive, well-written, sourcebook on the recovery 

and the forensic examination of tire track evidence. This book covers all aspects of 

tire impression examinations, from the design and manufacturing of tires, through 

the examinations at the scene and the documentation of the tire tracks there, to 

the examination at the lab and presentation in court. The book also includes 

information regarding tire tread databases, like the Tread Design Guide (by Tire 

Guides Inc., USA) and others. 

The recording of known test impressions from a suspect vehicle was discussed by 

several authors. Nause and Soulier (40) presented a simple and effective 

technique, based on the use of spray-on cooking oil, black fingerprint powder, and 

safety film, for the recording of tire impressions. This approach was found to be 

user-friendly, used easily obtainable materials, and gave excellent results. 

LeMay and colleagues (41) studied the effect of tire pressure and cargo weight on 

the width of tire track impressions. These authors made test impressions of tires 

with various air pressures and different weights of cargo in the vehicle in order to 

determine whether the width of a tire impression changes based on those 

variables. The results obtained suggest, not surprisingly, that the width of the 

contact patch varies according to tire pressure and weight of cargo. 

2.4 Test Impressions 

Preparing the right test impression, from the suspect’s footwear, is sometimes the 

basis for successful identification. Examiners occasionally find it necessary to 

59

make 3D test impressions of footwear when they are comparing the footwear to 

photographs of 3D crime scene impressions or to castings of these impressions. 

There are several products available for such use. Some are polymers that require 

mixing and hardening, and others are foam products that do not render fine detail. 

A new product, Bubber (Delta, Sweden), was recently tested by LeMay (42) and 

was found to be very easy to use and superior to other commonly-used products. 

It rendered very fine detail that could be photographed and cast with dental stone. 

This product may be used for preparing 3D test impressions from tires as well. 

2.5 The Evidential Value of Shoeprints Examination 

As discussed earlier in this Review, the identification of shoeprints is derived from 

individual characteristics found both in the suspects’ shoe outsole and in the crime 

scene impressions. In other, more frequent, cases, where no individual features 

can be identified, the examination conclusion is of the form “the examined 

footwear could have left the questioned impression from the crime scene”. In such 

cases, the evidential value of the results is often misunderstood and undervalued. 

LeMay (43) has recently discussed this issue, stressing the argument that even 

when only class characteristics (like outsole design, actual size and wear) are 

found, it is highly inclusive in nature, and must be treated as valuable evidence. 

Footwear impressions found at a scene are simply too far beyond coincidental to 

be dismissed. The example presented in this article is of a Nike athletic shoe, 

which was distributed in the USA in numbers exceeding 280,000 pairs (of this 

specific outsole thread design). That is less than 0.2% of all shoe pairs distributed 

annually in the USA at that time (44), while not yet taking into account the shoe 

size and wear. 

It should be mentioned at this point that popular footwear brands, like Nike, suffer 

from counterfeits of their products, thus the distribution figures presented by these 

firms do not necessarily represent the actual share of any given outsole design in 

the footwear population. A paper by Wisbey (45), which focused on the Nike Air 

Force One sneakers, highlights some of the methods that assist the shoeprint 

examiners assessing likelihood that a submitted sneaker may be counterfeit. 

Petraco and et al (46) applied statistical techniques used in facial pattern 

recognition, to a minimal set of information collected from accidental patterns in 

five pairs of similar footwear (all of the same make, model and size), in order to 

assess the evidential value of individual characteristics. In order to maximize the 

amount of potential similarity between patterns, these authors only used the 

coordinate locations of accidental marks to characterize the entire pattern. It was 

found that in 20–30 dimensional principal component (PC) space (99.5% variance 

retained), patterns from the same shoe, even at different points in time, tended to 

cluster closer to each other than patterns from different shoes. This study is 

intended to be a starting point for future research into building statistical models on 

the formation and evolution of accidental patterns. 

The Bayesian approach, advocated in the last decades by many forensic scientists 

for the application in various disciplines, has not yet been fully studied for footwear 

60

impression analysis. Biedermann and co-authors (47) discuss shortly the 

implementation of this approach into shoeprint examinations, and deal with 

feasible and defensible strategies for eliciting reasonable prior probabilities for 

footwear marks compared to DNA stains analysis. 

The range of conclusions, attributed to a given set of shoeprints, was presented by 

Jonasson (48). In the course of the ENFSI EWGM 2 nd Collaborative Shoeprint 

Test, participants were supplied with images of two footwear impressions, one on 

floor tiles and the other – on a curtain, and images of a pair of relatively unused 

tennis shoes and their test prints, and were asked to compare the prints with the 

shoes images. The conclusions reached by the participating experts ranged from 

elimination to identification, for the same given set of exhibits. This study 

demonstrates again, as did previous ones, the subjective nature of footwear 

impressions analysis. Based on the results of this test, a third one is being planned 

for distribution this year. 

2.6 Databases, Reference Collections and Automated Classification 

With the numbers of both footwear outsole designs as well as shoeprints 

documented at crime scenes rapidly increasing, the need for computerized mean 

of keeping these records is becoming more and more crucial in forensic 

laboratories. Bowen and Schneider (49) have published a review on various 

commercially-available forensic databases, including those for shoeprints and tire 

tread designs. 

In addition to the ENFSI EWGM “Wanted Page” mentioned earlier, Hamm (50) 

also presented the Foster & Freeman (UK) footwear and tire tread databases, 

including the Crimeshoe.com program. This “pay-per-match” program (customers 

only pay when a positive identification of the pattern in question have been found) 

enables law enforcement agencies and forensic science laboratories to submit 

queries regarding unknown outsole pattern, and receive a comprehensive report 

when such a pattern is identified in the database. 

The National Footwear Reference Collection (NFRC) has been launched recently 

in the UK (51), as the result of collaboration between the National Policing 

Improvement Agency (NPIA) and Bluestar Software Ltd. (UK), the latter having 

designed and built the database of footwear patterns. This new, web-based 

application service is hosted by the West Yorkshire Police on the secure Criminal 

Justice Extranet (CJX) and is freely available to police forces. Basically, the NFRC 

is a system for the identification of a particular footwear pattern using 14 

nationally-agreed descriptors of the different elements making up the design of the 

tread pattern. This allows police forces to identify footwear impressions recovered 

from crime scenes rapidly and easily. 

The potential of a spatial-temporal method for analysis of forensic shoeprint data, 

collected at the Larger London Metropolitan Area (the Bigfoot database), was 

examined by Lin et al (52). The large volume of shoeprint evidence recovered at 

crime scenes (about 10,000 annually) was imported into a geographic information 

61

system (GIS), and a spatial-temporal algorithm developed for this project. The 

results show that by using distance and time constraints interactively, the number 

of candidate shoeprints that can implicate one or few suspects can be 

substantially reduced. The study concludes that the use of space-time and other 

ancillary information within a geographic information system can be quite helpful 

for forensic investigation. 

Many authors have recently proposed systems for automated recognition, 

classification and retrieval of shoeprint patterns. Al-Garni and Hamiane (53) 

developed an efficient automatic shoeprint retrieval system based on Hu’s moment 

invariants. The performance of this algorithm is not significantly affected by 

decreasing the image resolution. It is also shown that the optimal performance of 

the proposed method is attained for rotated images. However, as stated by the 

authors, this system is suitable only for comparing suspect outsole patterns, and 

not partial shoeprints. 

Dardi et al (54) presented an image retrieval algorithm which combines the 

information of the phase of the Fourier transform of the shoe mark images with the 

power spectral density of the Fourier transform calculated on their Mahalanobis 

map. Different from other published studies, the algorithm performance here is 

tested on real shoe marks from crime scenes. The proposed method is compared 

with other studies and some preprocessing operators are also introduced and 

selected to reduce noise and enhance the matching probability. 

Pavlou and Allinson (55) developed an automated system for shoe model 

identification from outsole impressions taken directly from the suspect's shoes that 

can provide timely information while a suspect is in custody. The underlying 

methodology is based on large numbers of localized features located using 

maximally stable extermal region (MSER) feature detectors. These features are 

transformed into robust scale invariant feature transform (SIFT) descriptors with 

the ranked correspondence between footwear patterns obtained through the 

application of modified constrained spectral correspondence methods. The 

effectiveness of this approach is illustrated for a reference dataset of 374 different 

shoe model patterns, from which 87% first-rank performance and 92% top-eight 

rank performance are achieved. These authors were also involved in the 

development of the Immersive Forensics Ltd. (UK) Latent Image Markup and 

Analysis (LIMA) system, designed to provide a unified and intuitive environment 

for the treatment of forensic images, including shoeprint and tire track impressions 

(56). 

A technique for automatic shoeprint matching, using multiresolution Gabor feature 

map, was presented by Patil and Kulkarni (57). Gabor transform has been used to 

extract genuine textural features in a shoeprint image. The proposed technique is 

invariant to variations in intensity and rotation, and performs better compared to 

results obtained using power spectral density (PSD) features for full print images 

with rotation, intensity and mixed attacks at all the ranks. Performance of the 

algorithm has been evaluated in terms of recognition rate and cumulative match 

score for full prints and partial prints. The method was found to be robust to 

Gaussian white noise and salt–pepper noise. 

62

Other articles in this area of automatic shoeprint recognition, published outside the 

forensic science arena, were only available in abstract form. Among these worth 

mentioning are studies by Gueham and his colleagues (58-60), regarding 

automatic recognition of partial shoeprints using a correlation filter classifier and 

Fourier-Mellin transform, and automatic classification of partial shoeprints using 

advanced correlation filters for use in forensic science. 

2.7 Miscellaneous 

The issue of footwear size distribution, and even the size of a specific shoe, may 

also be important. An article by Turner (61), in the SATRA Bulletin, explains the 

different sizing scales and the nomenclature used by the footwear industry in this 

aspect. 

Identifying the make and model of footwear recovered from crime scenes is indeed 

a standard police technique. Taking it a step forward, Tonkin and his colleague 

(62, 63) demonstrated how this information could be used in the profiling of 

offender characteristics. Their findings demonstrate the value of footwear evidence 

as a prospective tool in the proactive search for offenders, which contrasts with the 

current use of footwear evidence in police investigations. Currently, the value of 

footwear evidence relies on the a priori identification of a suspect from whom to 

take an impression for comparison purposes. This, of course, limits the value of 

footwear evidence. However, the studied technique overcomes these limitations 

by allowing footwear evidence to help the police in their search for an unknown 

offender. 

Various materials adhering to the surface of footwear, or found inside it, are a 

potentially fruitful source of information for forensic reconstruction. Morgan and her 

colleagues (64) present a case where a soil sample was found on the suspect’s 

footwear, and compared with soil samples from the body burial site and from a 

suspect vehicle. It is interesting to mention that these authors also studied the 

persistence of soil residues on the shoes following machine wash, and found that 

even though the footwear looked clean, such residues were recovered from their 

inner part. 

Riding and his colleagues (65) studied the changes in pollen assemblages on 

footwear that had been worn at different sites. This study shows that when mixing 

occurs from wearing footwear at different sites, the pollen/spore content of the 

boots etc. dominantly reflects that of the last site. 

3. Toolmarks 

From a practical point of view, the field of toolmark examinations (namely the 

examination of marks produces by surfaces other than firearms) is, in a way, the 

“poor family-member” of firearm identification. Many of the professional groups, as 

well as published articles, are dedicated mainly to firearm examinations, and 

toolmarks are referred to matter-of-factly. Since firearm issues are covered by 

63

another review in this Symposium, this section of my Review will focus mainly on 

those aspects more relevant to toolmarks per-se. 

A general review of the procedures involved in firearm and toolmark identification 

and the validity of the results was published by Bunch and his colleagues (66). 

Following a description of the multi-level examinations performed by the examiner, 

the scientific foundation of this field and the way conclusions are being drawn, 

these authors concluded that the firearm-toolmark discipline is both highly valuable 

and highly reliable in its traditional methods. However, additional research is very 

beneficial and, depending on its purpose and design, would tend to better address 

potential error, identify manufacturing methods that are suspect for comparison 

purposes, and further develop machine systems and perhaps probabilistic models. 

The Scientific Working Group for Firearms and Toolmarks (SWGGUN) has 

recently published guidelines covering several aspects of firearm and toolmark 

examinations, like criteria for identification, training and quality assurance (67), 

which are relevant for toolmark examinations. These guidelines, like other 

SWGGUN documents, are publically-available on the Internet. On the light of 

recent challenges on the admissibility of various scientific evidence types in court, 

SWGGUN maintains an admissibility resources kit (ARK), assisting firearm and 

toolmark examiners prepare better for court appearances and face Frye, Daubert 

and other similar challenges. 

3.1 Casting and Reproduction Methods 

Many cases of toolmark examinations involve the need for duplicating the 

impression marks, found at the scene of crime, in order to facilitate their 

examination at the laboratory. Several materials had been proposed for this 

purpose, like silicone rubber and dental stone. 

Today, various suppliers of metallographic, dental and crime scene investigation 

products are offering silicone rubbers in easy-to-operate dispensers (dispensing 

guns), so avoiding the need for mixing the base with the hardener manually. 

Among these may be mentioned, for instance, the Forensic Sil (by Loci Forensic 

Products, The Netherlands), the Isomark products (UK), the Struers RepliSet 

(Denmark) and the AccuTrans (USA). These dispensing guns (AccuTrans) were 

evaluated by Watkins and Brown (68) and by Naccarato and Perersen (69), who 

found them to be both practical and accurate for most toolmark applications. 

Yo and his colleagues (70) compared five commercially-available casting 

materials, including four types of silicone rubber and dental stone, and evaluated 

their performance. Class and unique characteristics of the surface in question (a 

Canadian two-dollar coin – “Toonie”) and on the casts were examined visually. 

Their results demonstrated clearly that all tested silicone rubbers outperformed the 

dental stone for this purpose. Despite some minor differences found between the 

silicone rubbers, no definite recommendation is given regarding any of these 

products. 

64

Preparation of the appropriate toolmarks standards, using the suspect’s tool, is an 

issue as well. Toolmark test exemplars are usually produced by applying a tool's 

working surface to a piece of soft metal such as lead, since the lead will replicate 

the microscopic grooves of the tool surface without altering them. An alternative 

material for the preparation of such marks was presented by Petraco and coauthors 

(71), who proposed using commercially-available jewelry modeling waxes 

for this purpose. The replicas obtained are accurate, precise, highly detailed, and 

1:1 negative copies of the exemplar tool’s working surface. They reveal in fine 

detail the class characteristics, wear patterns, damage, and accidental markings 

present on a tool’s surface. 

Proficiency test samples are required to be identical, so each and every participant 

is analyzing or testing a similar sample. In order to produce test samples for the 

ENFSI Expert Working Group Firearms and Gunshot Residue (EWG FA-GSR), a 

procedure for a “mass-production” casting method for complex 3D objects (like 

bullets, cartridge cases etc.) was developed by Koch and Katterwe (72). 

Preparation of the test samples includes two stages: making the moulds (“negative 

casting”) of transparent silicone rubber (Elastosil by Wacker) followed by casting 

the samples into these moulds (“positive casting”). A detailed graphic description 

of the procedure is included in the article. Using this approach, it is possible to 

undertake proficiency testing in the field of firearm and toolmarks simultaneously, 

with identical samples for each participant. 

3.2 Observation and Imaging Methods 

The use of 3D imaging technologies for the potential application in forensic firearm 

and toolmark identification was evaluated by Bolton-King and her co-authors (73). 

A review of state-of-the-art profiling systems is provided, and particular attention is 

paid to the application of 3D imaging and recording technology to firearm 

identification. Each technology tested uses a different technique or scientific 

principle to capture topographic data, such as focus-variation microscopy, confocal 

microscopy, point laser profilometry and vertical scanning interferometry. In order 

to establish the capabilities and limitations of each technology qualitatively, 

standard reference samples were used and a set of specific operational criteria 

was devised for successful application in this field. The reference standard 

included the National Institute of Standards and Technology (NIST) 'Standard 

Bullet' in order to ensure that the evaluation represented the practical examination 

of ballistic samples. Based on this research, it was concluded that focus-variation 

microscopy is potentially the most useful approach for forensic examinations, in 

terms of functionality and 3D imaging performance. 

Ahvenainen et al (74) employed scanning white-light interferometry (SWLI) for 

comparing toolmarks produced by diagonal cutters on copper wires. There results 

suggest that SWLI may be applied as a quantitative method for forensic toolmark 

study through its high-resolution digital 3D profiles. 

The high depth-of-field required sometimes for toolmark examination (especially 

on rough and un-even surfaces), and the high magnification needed, led to several 

attempts for using scanning electron microscopy (SEM) for that purpose. Randich 

65

et al (75) discuss the advantages of using the SEM for examining deep-seated 

marks, and review the various imaging capabilities of this instrument. In order to 

show the usefulness of the proposed approach, SEM and conventional optical 

microscopy were used to examine firing-pin impressions in 9mm cartridge cases, 

and the power of the SEM was demonstrated. 

Katterwe and his colleagues (76) also describe the use of a comparison-SEM for 

forensic applications, and present the “next generation” of such instruments – the 

comparison variable-pressure SEM (VP-SEM). The conventional comparison SEM 

comprises of two SEMs linked together trough a mutual control system, so that the 

two instruments are synchronized electronically into a single video screen. The 

comparison VP-SEM overcomes one of the limitations of the conventional SEM, 

which is the need for conducting samples, thus enabling the relatively simple (but 

quite expensive, NL) examinations of non-conductive specimens (made of plastic, 

wood etc.). 

In his previously-mentioned report, Prokoski (17) also studied the application of an 

infrared (IR) imaging system for examining and matching of toolmarks created by 

screwdrivers. The resulting marks were imaged using an IR camera with 640x512 

detector array and 3X optics and also a visible light 2592x1944 CCD camera. The 

IR images produced more consistent striations than the visible light images. The 

study goal was to demonstrate whether IR was superior to visible light imaging 

with respect to accuracy of match for 20 toolmarks against the database of 200 

marks. The method for matching striations was a comparison of “barcode” 

representations of "cut-lines" through the toolmark perpendicular to the direction of 

the mark. As a result, each of the 20 IR secondary marks was correctly matched to 

the primary mark made by the same tool. 

3.3 Marks Produces by Various Types of Tools 

Since the evidential value of toolmark examinations rests on the “uniqueness” of 

the mark producing surface, knowledge of the manufacturing processes of various 

types of tools is an essential part in these examinations. 

Lang and Klees (77), while discussing the broader issue of forensic drill bit 

examinations, including use indicators, swarf (cuttings) composition, etc., deal also 

with the toolmarks left by these items. Two types of marks may be encountered 

when drilling: marks on the work piece (especially in partially-drilled holes), and 

those found on the remaining swarf. The reproducibility of toolmarks on certain 

types of swarf is presented, and the application of these examinations in 

improvised explosive devices (IEDs) cases is demonstrated. 

Marking stamps (punches) are used for producing imprints, usually on metal 

surfaces. Comparing such marks with the suspect stamps may have important 

value in cases like forgeries of vehicle chassis and engine numbers. Weimar and 

his colleagues (78) describe in detail the manufacturing processes involved in the 

production of these punched, and the implications of each method on the expected 

toolmarks produced. These authors conclude that the punches produced by the 

66

three manufacturing processes examined show identifying characteristics 

immediately after the production. In some cases it cannot be decided if such 

characteristics are identifying or class characteristics, e.g. in the case of 

impressions in the indentation edge if only one punch is available from a 

production series. Punches produced by milling and cold forming exhibit fine and 

complex structures (milling grooves, striation marks and wave-like structures 

transferred from the die-head) which are not identifying characteristics because 

they appear on several surfaces of punches produced in one consecutive series. 

Drug packaging is often being submitted for examination in forensic science 

laboratories. Most common examinations in such cases are fingerprints or even 

physical match, but toolmarks or manufacturing process marks may also be 

examined. Dutton (79) presented a case where morphine tablets packaging (foilbacked 

plastic blister packs), found in the possession of two separate suspects, 

were linked together by using both manufacturing marks and physical match. 

Firearm examiners are also faced, from time to time, with marks originating from 

surfaces other than firearms per-se. Haag (80) discuss an old-argued question of 

linking cast bullets to the mould that made them. Earlier published works had 

raised the issues of carry-over and subclass characteristics on bullets from 

consecutively manufactured bullet moulds. Based on the findings of the current 

study, the presence of unique damage or obvious individual characteristics can 

allow an accurate association of cast bullets to their mould. 

Other manufacturing marks on pieces of ammunition, this time – the headstamp 

impressions on cartridge cases, were studied by Tidrick and co-authors (81). In 

contrast to previous research, that has shown that bunter toolmarks, which are 

transferred to each headstamp, are unique and persist for a considerable part of 

the production run, this article suggest that bunter toolmark examinations are not 

particularly useful for associating or disassociating cartridge cases. According to 

the results obtained, at least two different bunter tools were represented in a box 

of ammunition and, within a box, a bunter tool may be represented by as few as 

one or two cartridges. Often, boxes from the same lot feature common bunter 

toolmarks, but identical bunter toolmarks were found in boxes of ammunition from 

different lot numbers. Also, identical bunter toolmarks were found in boxes of 

ammunition bought in different locations. 

Miller (82) reports a case in which it was confirmed that an axe blade was 

responsible for producing the marks evident on a damaged door. The application 

of the blade at the time the toolmarks were made was determined, and a jig was 

specially designed to assist in reproducing the toolmarks as closely as possible to 

the manner in which they were produced when the offense was committed. 

Consideration is given to the types of toolmarks, tool action, and comparison 

techniques employed. 

Burda and her co-authors (83) examined cable ties of different make and sizes, 

and found that these items may bear manufacturing-processes marks (mould 

details, ejection pins details, etc.) that can be used for comparison between 

suspect and known ties. 

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3.4 Examination of Consecutively-Manufactured Tools 

One of the ways of demonstrating the uniqueness of toolmarks is by studying the 

marks produced by consecutively-manufactured tools or firearms. Buckleton and 

his colleagues (84) suggested an experimental design for acquiring relevant data 

for addressing this issue. The authors accept the fundamental soundness of this 

approach, and do not suggest that previous work was unsound, but urge the 

toolmarks examiners’ community to adopt several modifications in the 

experimental procedure: “Blinding” the test, distribute the exhibits to the examiners 

randomly, and publish all the results . The proposed experimental design is a lot 

more time consuming than the usual experiment, however these authors feel that 

many of the minor faults in current experiments would be eliminated by this 

approach. 

An example for an empirical study for the validation of toolmarks examination was 

published by Giroux (85). Five consecutively-manufactured flat-bladed 

screwdrivers were acquired from the manufacturer, and toolmarks were produced 

on sheet-lead test material, in a uniform manner, using both sides of each tool. 

The test samples were then randomly divided into test-sets (kits) and examined by 

eight qualified toolmarks examiners (each received, randomly, one known mark 

and 10 questioned ones). As published, there were no mis-identifications in this 

study – 0% false positive error rate (0/51). However, there was one mis-elimination 

– 3.4% false negative error rate (1/29). This mis-elimination calls for attention to 

the topic of elimination criteria, especially when the questioned tool is not available 

for examination. 

Another study regarding the toolmarks produced by consecutively-manufactured 

screwdriver tips was conducted by Chumbley, Faden and co-authors (86-88), as 

part of a project on the development of a non-destructive optical profilometry 

method for toolmark examination. The profiles obtained were compared and 

statistically evaluated using an algorithm developed for comparing twodimensional 

images of toolmarks. According to these authors, their results 

demonstrate that screwdriver tips are unique in surface profile, not only from tip to 

tip, but from side to side of each tip as well. 

As part of research into the validation of this analysis, initiated by relevant US 

court decisions (including, for instance, Ramirez vs. State of Florida, No. 66,992, 

1989), Lancon (89) examined marks made in bone using ten consecutively 

manufactured knives. The test cuts in the bone (pig ribs) and the reference cuts 

were all recorded using silicone-based casting material (Microsil®), and compared 

using a comparison microscope. Eventually, all of the blades could be matched to 

their corresponding cuts. Representatives of the best known non-match (KNM) 

comparisons were photomicrographed for further reference. According to the test 

results, bone is a suitable material for accepting toolmarks, and consecutively 

manufactured knife blades can be differentiated using conventional techniques 

used by firearm and toolmark examiners. 

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3.5 The Examination of Stabbing and Cutting Marks 

Many of the studies, conducted in the Review period in the field of identification 

and comparison of marks, deal with stabbing, cutting or sawing marks to the 

human body. Saville et al (90) studied saw marks on bone using a VP-SEM 

(Environmental SEM – ESEM, FEI, The Netherlands) and found three levels of 

striations on the kerf walls and floors, each one attributed to a different area of the 

cutting saw motion. Two of these striation levels (Types A and B), visible using 

stereomicroscopy, are the result of the pushing and pulling of the saw, and they 

can provide class characteristics of the saw (its width, the number of teeth per inch 

- TPI, etc.). According to these authors, the third group of striae (Type C), 

observed here for the first time, are produced by each individual tooth, and may be 

used for individualising the specific saw that had left them. The “uniqueness” of 

these Type C striations is discussed in the article, and the conclusion drawn by 

these authors is that they are unique, based on the results of a limited blind test. 

The observation of these Type C striations is only possible by using the ESEM, 

due to this instrument high magnification and depth-of-field and its ability to deal 

with non-conductive and wet samples. 

VP-SEM for the analysis of saw marks in bone was employed also by Freas (91). 

This author compared the images obtained using light microscopy with those 

obtained using the VP-SEM, and studied the wear-related changes in the kerf wall 

and their impact on the interpretation of saw marks in bone. Sequences of 30 cuts 

in bone were produced using crosscut saw and hacksaw, revealing patterns of 

progressive loss of fine details of kerf wall morphology with increasing saw blade 

wear, because of the rounding of sharp points and edges. Nevertheless, 

diagnostic kerf wall features used to establish class characteristics persist despite 

these wear-related changes. Statistical analysis of wear-related changes, based 

on striae width and density, was found to be unsuccessful, suggesting these 

patterns are not readily quantifiable. According to this study, despite the scanning 

electron microscope’s superior imaging capabilities, it provided only few practical 

and methodological gains over the traditional light microscopy. 

Marciniak (92) examined the degree of modification of saw-mark characteristics of 

dismembered skeletal remains exposed to an outdoor fire of limited duration. The 

study material consisted of 36 adult pig hind limbs which had been dismembered 

in a fleshed condition. Six different handsaws and six power saws were used, with 

three limbs dismembered by each saw type prior to exposure to fire. According to 

the results obtained, exposure to fire influences the visibility and identifiability of 

the saw-mark striations, but it is still possible to identify the class of saw used on 

the basis of the characteristic marks present on the cremated bones. 

Bailey and his colleagues (93) studied the kerf marks made by several types of 

hand- and mechanical-saws, in order to assess the ability of microscopic 

measurements of the kerf mark width to differentiate the tools that produced them. 

Their results suggest that this phenomenon may be used as an effective measure 

for eliminating some saw blades. 

Using the SEM, Lynn and Fairgrieve (94) studied the trauma to mammalian long 

bones by axes and hatchets. It was found that striations were present on the 

69

smooth impact sites of fleshed and defleshed bones, contradicting the findings 

previously published results. The consistently smooth impact sites and rough 

fracture surfaces may be useful features in the characterization and reconstruction 

of axe and hatchet wounds. 

Wong (95) searched for the most appropriate method of preserving and examining 

tool marks on cartilage and bone. Guillotine paper cutter was used to cut the 

cartilage specimens, while a fire axe was used to chop the bones. Ten different 

combinations of preservatives and storage conditions were also compared. Casts 

of the tool-marked surfaces, made before preservation, served as a baseline for 

comparison. The casts made after preservation were subjected to microscopic 

examination, compared to the pre-preservation casts, and scored to evaluate the 

performance of the preservatives in their respective storage conditions. The 

results obtained suggested that most aqueous solutions stored in a cold 

environment were good at preserving the finer accidental tool mark characteristics. 

It was concluded that the optimal method involved the immersion of the samples in 

a 0.9% NaCl saline solution, frozen storage, air-drying after removal from storage, 

and casting with rubber-base polysulfide dental impression material (COE-FLEX 

®) for identification work. 

The dynamics of knife punctures in cervical vertebra and in tire rubber was studied 

by Locke (96). In the course of a murder investigation, a cervical vertebra of the 

victim was examined. It was shown that toolmarks present on the cervical vertebra 

of the victim had been made by one edge of the swedge of the suspect knife - a 

finding that led to further research into the dynamics of a knife puncture. This 

research showed that the characteristic “Y” tear observed in a tire puncture is 

more than a tear, being two cuts made by the left and right edges of the back of a 

wide-blade knife. 

Bearing in mind the paucity of articles in the area of toolmarks on bone, Fred 

Tulleners of the University of California Davis has reprinted a 1947 article by 

Thomas and Gallent (97) regarding the identification of the weapon used in a 

murder case. By comparing the toolmarks made by the axe (found in the 

possession of the suspect) with the marks made by the weapon on the skull of the 

victim it was determined that the axe had been used to commit the offence. 

Interestingly, the 1947 includes even statistical evaluation of the results. Other 

articles dealing with the characterization of trauma caused by various types of 

tools were found to be outside the scope of this Review. 

3.6 Evidential Value of Toolmark Examination 

Toolmark (as well as firearms identification) examinations have gone through the 

same scrutiny as other classic identification areas. The way forensic scientists are 

taking the “leap of faith” (98), stating that a questioned mark was made by a 

specific known tool (with the exclusion of all others), drew a lot of criticism, mainly 

from non-forensic-scientists (99-101, and others). In her commentary on Nichols 

(102) extensive article, Schwartz (103) argued again that the AFTE Theory of 

Identification is not a valid one, since it is based on the limited experience of each 

examiner, that proficiency tests are usually easier and less complicated than 

70

casework samples, and that the lack of statistical data (unlike DNA analysis) 

undermines the evidential value of the examination. In addition, Schwartz claims 

that there are no enough studies regarding the differentiation between subclass 

and individual characteristics. 

In other articles, addressed to defence attorneys, Schwartz (104, 105) suggests a 

line of questioning for cross examination of firearm and toolmark experts in court. 

These articles are useful reading material for any forensic scientist in the field of 

toolmarks examination, in particular when preparing for a court testimony. 

As mentioned earlier, differentiating between known matches (KM) and known 

non-matches (KNM) is particularly important because of the legal challenges that 

examiners may face in court. Neel and Wells (106) performed a study in order to 

quantify the difference between KM and KNM, for better understanding of these 

definitions. Various toolmark sources were used, including two- and threedimensional 

toolmarks. More than 4,000 striated toolmark comparisons were 

examined for their Total Matching Lines (TML), Percent Matching Lines (%ML), 

and Consecutively Matching Striae (CMS). The results obtained show that 2D and 

3D KM and KNM are statistically distinguishable from one another. 

Bachrach et al (107) report the application of confocal microscopy and white-light 

interferometry for the acquisition of 3D (topographical) data of a considerable 

number of striated toolmarks created under controlled conditions on a variety of 

media. Toolmarks were produced using screwdrivers and tongue-and-groove 

pliers. The obtained toolmarks were then evaluated using algorithms, and the 

distributions of the degree of similarity values obtained from the comparison of 

known matching and nonmatching pairs of marks were analysed using established 

statistical technique. Empirical error rates were calculated as a metric of tool mark 

individuality, where a low empirical error rate is indicative of high specificity and 

repeatability. While it is not possible to prove uniqueness statistically (98), the 

results of this study provide support for the concept that toolmarks contain 

measurable features that exhibit a high degree of individuality. 

During the aforementioned study, Chumbley et al (88) compared the results 

obtained by using computerized analysis with those of toolmarks and firearms 

examiners. During the 2008 AFTE Training Seminar, volunteers were solicited into 

participating in the examination of the same toolmarks earlier evaluated by the 

algorithm. The AFTE volunteers were asked to compare those samples that were 

found to be most difficult for the computerised algorithm, but still no false positive 

results were reported by any of the AFTE volunteers. They did, however reported 

some false negative results, mainly in a specific pair of samples, probably due to 

the fact that the tools were not available for examination. These finding are in 

agreement with other reported studies (85, for instance). 

An interesting study regarding the calculation of the theoretical significance of 

matched bullets, but having impact on toolmark analysis as well, was published by 

Howitt and co-authors (108). These authors present the derivation of the formulae 

for calculating the probability for the correspondence of the impression marks on a 

subject bullet to a random distribution of a similar number of marks on a suspect 

71

ullet of the same type. This involves the subdivision of the impression marks into 

a series of individual lines with width equals to the separation distance at which a 

misalignment of striations between the bullets is indistinguishable. This distance 

depends on the microscopic resolution limits and the visual acuity of the examiner. 

Calculated probabilities for finding pairs and triplets of consecutively matching 

striations on non-matching bullets, by an examiner with normal eyesight using a 

microscope at 40X magnification, gives values that are in good agreement with the 

empirical probabilities determined in the 1950s, and when determined for larger 

consecutive sequences indicate that they are very unlikely to occur. These 

formulae can be used to calculate the probabilities for the random occurrence of 

any sequence of striations, providing a means by which the significance of a 

specific match between any two bullets can be justified quantitatively. 

The error rates of any given scientific evidence form today a substantial element in 

the admissibility of this evidence in court. Murphy (109) addressed this issue in a 

presentation available at the SWGGUN Admissibility Resource Kit (ARK), based 

on the Collaborative Testing Services (CTS) firearm and toolmark examination 

tests (1992-2005). Despite the views expressed by the tests’ provider, CTS, that 

“CTS Summary Reports should not be used to determine forensic science 

discipline error rates…” (110), Murphy analyses the data anonymously published 

in the CTS final reports and presented the calculated false-positive and falsenegative 

rates, as well as the sensitivity and the specificity of both firearm 

identification and toolmarks examination. The calculated error rate for the 

toolmarks proficiency tests are as follows: False positive (wrong identification) rate 

– 1.7%, false negative (wrong exclusion) rate – 1.6%, sensitivity (the ability to 

detect the right identification) – 90.6% and specificity (the ability to detect the right 

exclusion) – 57.9%. Since one of the objections for using the CTS reports for error 

rates evaluation is that not all of the participants of these tests are qualified 

experts, Murphy presents also the error rates when the trainees removed, and find 

them to be even lower. 

3.7 Miscellaneous Issues 

Swanepoel (111) presented an interesting and rarely-published case where unique 

dual-impression encountered during the comparison of a stolen hydraulic pump 

and the base plate from which the pump was allegedly stolen. There was an 

agreement of class characteristics and sufficient agreement of individual 

characteristics which were of such significance that it could be concluded that the 

hydraulic pump and the base plate were at one time connected to each other. 

The forensic analysis of knot evidence is an uncommon examination type. 

Nevertheless, Chisnall published several articles in this field, dealing with the 

strength and limitations of such analysis, with tying anomalies and their 

significance in analysing knot evidence and with tying habits (112-114). This 

author describes how properly preserved and analysed knot evidence can offer 

corroborating information, indicate leads to other sources of evidence. A survey of 

562 volunteers, conducted by Chisnall, revealed that only a minority ties 

noteworthy anomalies that differ from the common trend. Finding these rare 

anomalies in crime scenes might have significant evidential value. If similar 

72

anomalies can be found in suspect samples, the link between the suspect and the 

case knots is stronger. In addition, the study shows that right-handed subjects 

tended to tie S-twisting hitches more often than Z-twisting hitches (S- and Ztwisting 

refer to the direction of one strand over the other) while left-handed ones 

demonstrated S tying less frequently and even tied Z knots more often in some 

situations. 

Desiderio and Chin (115) stress the need for a more synergistic approach for 

toolmark examinations, and are discussing the important role that trace evidence 

(like physical match, paint or DNA) plays during the examination of toolmarkrelated 

evidence. In order to illustrate the significance of the relationship between 

trace evidence and toolmarks, being complementary to each other, various case 

related studies are presented. 

4. Physical Match 

Physical match, namely linking two or more objects by the morphology of fractured 

or torn surfaces, is usually viewed as one of the strongest ways for establishing 

common origin. The evidential value of such physical matches, and their 

admissibility in court, seem to be taken for granted, considering the limited number 

of articles published during the Review period in this discipline. 

Christensen and Sylvester (116) conducted a validation study for the reliability of 

physically matching fragments of bone and other mineral-based biological 

materials such as shells and teeth. Participants with varying education, training 

and experience were asked to complete a matching exercise consisting of 

intentionally fragmented specimens. Success rates were very high - the positive 

association (correct match) rate was 0.925, while the non-association (overlooked 

match) rate was 0.075, and negative associations (incorrect matches) occurred at 

a rate of just 0.001. Results also indicate that participants with more education and 

related experience tended to have higher positive association rates, although not 

significant statistically. Experienced osteologists, however, completed the 

matching exercise in significantly less time. Low error rates among both 

experienced and inexperienced individuals support the reliability and validity of 

performing physical matches of these materials, and suggest that performance 

may also be related to an individual's aptitude for spatial tasks or other factors. 

In a series of publications, De-Smet and his colleagues (117, 118) presented two 

dimensions (2D) and three dimensions (3D) fracture matching of snap-off cutter 

blades, using numerical algorithms and surface area based reliability evaluations. 

Like other researchers, these authors used controlled breaking conditions, using a 

materials testing machine, for producing the test samples. The use of a 

commercial white-light profilometer system for obtaining 2D and 3D image surface 

scans of multiple fractured objects is discussed, and the results showed that this 

approach performs quite well, confirming all matches in the test-set and passing a 

blind test. The conclusions of this project were that 3D profilometry can aid in 

developing more objective methods for examining fracture matching surfaces, that 

evaluation of automated methods parameters may be a required step towards 

73

proper probability based fracture match reasoning and reporting, but also that 

automated methods should not be used or applied blindly. 

In a yet-unpublished report, following Tsach et al (119), Yekutieli et al (120) 

demonstrated a prototype system used for physical matching in 2D. The system 

has two main functions: One is to assist forensic experts in performing physical 

matching in an objective manner, and the second - collecting statistics and build 

confidence levels regarding physical matches. The probability distribution 

functions (PDFs) of matching error values, for correct matches and for nonmatches 

were estimated. This analysis was applied for different fracture line 

lengths and three different materials. Eventually, these authors were able to 

calculate error rates much more reliably than previous estimates. With the results 

of this research, an expert can express his findings in a more numerical way, and 

the Daubert criteria for a potential or known error rate can be fulfilled. Surprisingly, 

statistical results were much lower than initially expected, probably because these 

authors used only the 2D fracture lines and not any additional information 

commonly used in fracture match comparison, such as the 3D nature of some 

fractures or any existing texture and graphic patterns on the surface or outer 

border of the pieces to be compared. 

As experience shows, the nature of the broken or the torn object can influence the 

results of the match. When examining the torn edges of plastic tapes, differences 

between the tape ends are sometimes observed, due to plastic deformation. 

Weimar (121) studied this phenomenon, and developed a method of overcoming it 

by heat treating the torn ends. According to this author, heat treatment of the 

examined PVC tapes made it easier to find corresponding tear edges, and the 

conclusiveness of the found match was increased. Because the vast amount of 

available different tapes, it is recommended to test the influence of heat treatment 

on tapes encountered in each case, while taking into consideration other types of 

evidences that might be influence by such treatment (fingerprints, DNA etc.). 

This type of evidence is extremely useful in the investigation of road accidents, 

especially in Hit-and-Run ones. Christophe and Daniels (122) presented a case of 

matching a wood chip found at the scene with a wooden pallet that had been on 

the back of the suspect's pick-up truck. The authors used Adobe Photoshop CS2 

software for superimposing the images of the two exhibits. Based on their 

examination of the surface contours of both items, a positive identification of the 

suspect’s vehicle with the accident scene was established. 

Other application of physical match may be in question document examination. 

Guscott (123) study a case of analysing several threat letters, one of which aimed 

at the local police department. Forensic document examination of the initial threat 

letter established a link between the multiple cases in the neighbourhood. This 

particular case involved multiple aspects of forensic document examination, 

including handwriting identification, indented writing, the discovery of an ink defect 

in the printing process of a notepad and a crease in another notebook, and a 

physical match that proved the connection between the different cases. The 

results of the forensic document examinations proved vital in securing a 

conviction. 

74

Another case of document examination involving physical match was presented by 

Brown and Sin-David (124), dealing with the remains of the late Israeli astronaut, 

Colonel Ilan Ramon, crew notebook, found in NASA’s Space Shuttle Columbia 

crash site. One of the methods applied while studying these exhibits was 

physically matching torn pieces of paper found in deferent locations. It should be 

noted in this case that the remains had undergone traumatic conditions and tears 

of the notebook pages did not show perfect physical matches as expected for 

paper torn under laboratory conditions. 

5. Restoration of Obliterated Marks 

Visualization of obliterated serial numbers, in chassis and engine of vehicles, in 

firearms frames and in other objects, may provide important forensic evidence 

during criminal investigations, when stolen items have their serial numbers 

obliterated in an attempt to conceal their identity or origin. Several articles have 

been published during this Review period, regarding methods dealing with this 

issue. In addition, updated information and procedures may now be found on the 

Internet (4, for example). 

The methods applied for such examinations may be divided usually into two 

groups (125): Non-destructive methods (like magnetic particles, Eddy current or xray 

radiography), and destructive methods (like chemical and electrochemical 

etching or thermal annealing). The appropriate method, or combination of 

methods, suitable for each surface, is dependent mainly on the surface 

composition and manufacturing history, on the marking methods and on the 

obliteration process. Many metallurgical tests may be required for each type of 

exhibit, for determining the proper procedure to be used. 

Weber and Weimar (126) proposed an approach for numerically simulating the 

marking process, thus reducing the number of actual metallographic experiments 

needed for developing a method for the marking restoration. The finite element 

method (FEM) simulation results were compared with actual metallographic 

experiments, with comparable results. These authors showed that the FEM 

simulation of the marking process is possible, and that the results of such 

simulations can be applied to metallographic examinations. 

Collaborative Testing Services (CTS) has just recently distributed their first 

proficiency test on the restoration of obliterated marks (Test No. 10-525). The final 

report, available over the Internet, includes a list of methods used by the 

participating laboratories (127). It is interesting to see that many laboratories used 

the magnetic particles method, prior to, or instead of, using chemical etching. 

5.1 Steel surfaces 

One of the most active groups in research in this field is led by Prof. Kuppuswamy, 

at the School of Health Sciences, Universiti Sains Malaysia. Following the work by 

Zaili, Kuppuswamy and Harun (128), reviewed in our previous Review (1), Yin and 

Kuppuswamy (129) studied several common reagents for the restoration of 

75

obliterated marks on medium carbon steel. The 2009 study have revealed that 

Fry’s reagent, comprising of cupric chloride (CuCl2), hydrochloric acid (HCl) and 

water, provided the necessary contrast and was concluded to be the most 

sensitive of the tested reagents. The same reagent was recommended by earlier 

workers for revealing strain lines in steel surfaces. Earlier, another reagent 

containing copper sulphate (CuSO4), water, concentrated ammonium hydroxide 

(NH3OH) and concentrated HCl was proved to be more sensitive for restoring 

erased marks on low carbon steel (128). 

Great minds probably think alike, since similar results, regarding the effectiveness 

of the Fry’s reagent, were also reported by Wightman and Matthew (130). These 

authors have also developed an etching paste, for use on steel surfaces (131). 

Their proposed paste is based on a mixture of alumina powder and the Fry’s 

reagent. The paste proved to be as effective as liquid in most cases, and often 

gave even better results. The paste is thixotropic, making it much easier to use, 

particularly with irregular shapes or on site. Coverage is good as vibration allows 

the paste to flow and give an even cover. Apart from cases where erasure has 

occurred by metal burrs filling the indentation, the paste gave as good a recovery 

as the liquid etch, and often it appeared to give better recovery. According to these 

authors, the ease of using the paste gives it distinct advantages over the liquid 

reagent. It seems, however, to this Reviewer (NL) that an apparent drawback of 

such paste is that it is not translucent, so the restored marks are not visible during 

the process, unlike when using the liquid reagent. 

Kamila and Colleagues (132) demonstrated the application of visual and 

microscopic, non-chemical, methods for the successful restoration of an 

obliterated serial number on a .38 calibre revolver submitted for examination. The 

conclusion rightly drowns by these authors is that visual and microscopic 

examinations, in tandem with other methods or independently, should always be 

applied in such cases. 

5.2 Aluminium alloy surfaces 

A recent work by Bong and Kuppuswamy (133) assessed the etching technique 

for the restoration of obliterated engraved marks on high-strength aluminium alloy 

(AA7010) surfaces. The aluminium surfaces were engraved mechanically with 

identification marks before the marks were erased by removal of the metal to 

different levels up to and below the depth of engraving. Five different 

metallographic reagents were then tested on the obliterated surfaces by etching. 

According to these authors, the most effective methods for the restoration of the 

obliterated marks were: (1) immersion in 10% aqueous phosphoric acid, and (2) 

alternate swabbing of 60% hydrochloric acid (HCl) and 40% sodium hydroxide 

(NaOH). Both procedures could also show the marks obliterated by overengraving 

and centre-punching. Notably, alternate swabbing of HCl and NaOH 

proved to be the common reagent for restoration on pure aluminium surfaces as 

well as on its alloys, providing support for the findings of previous studies. These 

findings are relevant because of the increasing use of high-strength aluminium 

alloys in car and firearm manufacturing. 

76

In a previous work, Baharum et al (134) studied the characteristics of restoration of 

obliterated engraved marks on aluminium surfaces by etching techniques, and 

found also that the alternate swabbing of HCl and NaOH the most sensitive one 

for these metal surfaces. This reagent was able to restore marks in the above 

plates erased down to 0.04mm below the bottom of the engraving. The marks also 

presented excellent contrast with the background. This reagent was further 

experimented with similar aluminium surfaces, but of relatively greater thickness of 

1.5mm. These authors noticed that the recovery depth increased slightly to 

0.06mm, suggesting the dependence of recovery depth on the thickness of the 

sheet metal. Further, the depth of restoration decreased in cases where the 

original number was erased and over-engraved. These results are similar to those 

of steel surfaces reported earlier. 

Peeler et al (135) also used a combination of acid etchant (HCl, 60% v/v in water) 

followed by an alkali one (NaOH, 40% w/V in water) for the restoration of 

obliterated numbers on aluminium alloy motorcycle frames. These authors 

recommendation is to repeat the acid-alkali cycle till the desired restoration is 

achieved. This method was successfully applied in casework. The safety issues of 

using strong corrosive acidic and alkali reagents are also discussed. 

5.3 Laser engraved marks 

A case of recovering obliterated laser engraved serial numbers in firearms frames, 

made of aluminium alloy, was presented by da-Silva and dos-Santos (136). 

According to this article, the traditional recovering methods of using acid etching 

generally fail in such cases, since the marking the serial numbers by laser 

engraving does not necessarily imply a deep permanent deformation of the 

crystalline array. The method applied here was manual relief polishing, coupled 

with reflected light stereomicroscopy and digital video photography. The manual 

polishing is a critical stage of the method, since the strength and the velocity of the 

process is closely related to the success of the method. 

Klees (125), while trying to pursuit novel methods for this field of investigation, 

studied the application of scanning electron microscopy, combined with energy 

dispersive x-ray spectroscopy (SEM/EDX, SEM/EDS) for restoring obliterated 

laser-engraved serial numbers on firearms. His experiments included both 

secondary electron (SEI) and backscattered electron (BEI) imaging, and x-ray 

mapping. The results presented in this work were not promising, and SEM/EDS 

was found to be ineffective for restoration of obliterated laser-engraved serial 

numbers. 

5.4 X-Ray Radiography 

Jeon and et al (137) describe the examination of vehicle license plated, where the 

original markings had been erased by hammering-off and new figures were 

embossed. For revealing the original marking on the plates, these authors used 

non-destructive x-ray radiography, followed by computerized image processing. It 

proved to be an efficient way for visualizing the hidden original figures on 

aluminium plates. 

77

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80

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81

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(89) Lancon DS. Toolmarks in Bone: Continuing Research with Consecutively 

Made Knife Blades. AFTE Journal, 2009, 41(2): 130-137. 

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the "Uniqueness" of Saw Marks on Bone. International journal of Legal 

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(94) Lynn KS and Fairgrieve SI. Microscopic Indicators of Axe and Hatchet 

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(95) Wong DT. Preservation and Examination of Tool Marks on Cartilage and 

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(98) Stoney DA. What Made Us Ever Think We Could Individualize Using 

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(104) Schwartz A. Challenging Firearms and Toolmarks Identification – Part One. 

The Champion, 2008 October, 10-19, 

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60e02deb31e6e8525754b0077e022?OpenDocument&Highlight=0,toolmark 

. 

(106) Neel M and Wells M. A Comprehensive Statistical Analysis of Striated Tool 

Mark Examinations Part I: Comparing Known Matches and Known Non- 

Matches. AFTE Journal, 2007, 39(4): 176-198. 

(107) Bachrach B, Jain A, Jung S and Koons RD. A Statistical Validation of the 

Individuality and Repeatability of Striated Tool Marks: Screwdrivers and 

Tongue and Groove Pliers. Journal of Forensic Sciences, 2010 March, 

55(2): 348-357. 

(108) Howitt D, Tulleners F, Cebra K and Chen SA. Calculation of the Theoretical 

Significance of Matched Bullets. Journal of Forensic Sciences, 2008 July, 

53(4): 868-875. 

(109) Murphy D. CTS Error Rates, 1992-2005, Firearms/Toolmarks. 2010, 

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(111) Swanepoel J. Physical Matching as Duties of a Firearms and Toolmark 

Examiner. AFTE Journal, 2007, 39(3): 215-226. 

(112) Chisnall R. What Knots Can Reveal: The Strengths and Limitations of 

Forensic Knot Analysis. Journal of Forensic Identification, 2007 September, 

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(113) Chisnall R. Tying Anomalies and Their Significance in Analysing Knot 

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Tool Mark Examinations. Forensic Science Today, 2008 April, 5: 2-9 

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Physical Match 

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(117) Hollevoeta D, De Smetb P, De Bockc J and Philips W. Towards Automated 

Forensic Fracture Matching of Snap-off Blade Knives. Interferometry XIV: 

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(118) De Smet P. 2D/3D Fracture-Matching of Snap-off Cutter Blades Using 

Numerical Algorithms and Surface Area Based Reliability Evaluations. 

Science & Justice, 2010 March, 50(1): 39 (abstract). 

(119) Tsach T, Wiesner S. and Shor Y. Empirical Proof of Physical Match: 

Systematic Research with Tensile Machine. Forensic Science International, 

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TP-2558 Final Report, NIJ, 2008. 

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Improvement of the Conclusiveness by Heat Treatment. AFTE Journal, 

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(122) Christophe DP and Daniels C. An Unusual Technique for Physical Match 

Comparison. AFTE Journal, 2008, 40(4): 396-398. 

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Restoration of Obliterated Marks 

(125) Klees GS. The Restoration or Detection of Obliterated Laser-Etched 

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AFTE Journal, 2009, 41(2): 184-187. 

(126) Weber M and Weimar B. Analysis of the Marking Process Using the finite 

Element Method. AFTE Journal, 2009, 41(2): 167-175. 

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(128) Zaili MAM, Kuppuswamy R and Harun H. Restoration of Engraved Marks 

on Steel Surfaces by Etching Technique. Forensic Science International, 

2007 August, 171(1): 27-32. 

(129) Yin SH and Kuppuswamy R. On the Sensitivity of Some Common 

Metallographic Reagents to Restoring Obliterated Marks on Medium 

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(130) Wightman G and Matthew J. Restoration of Stamp Marks on Steel 

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(131) Wightman G and Matthew J. Development of Etching Paste. Forensic 

Science International, 2008, 180: 54-57. 

(132) Kamila GH, Abraham JT and Bhattacharyya CN. Obliterated Firearm Serial 

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(133) Bong YU and Kuppuswamy R. Revealing Obliterated Engraved Marks on 

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Forensic Science International, 2010 February, 195(1-3): 86-92. 

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(134) Baharum MIM, Kuppuswamy R and Rahman AA. Recovering obliterated 

engraved marks on aluminium surfaces by etching technique. Forensic 

Science International, 2008, 177(2-3): 221-227. 

(135) Peeler G, Gutowski S, Wrobel H and Dower G. The Restoration of 

Impressed Characters on Aluminium Alloy Motorcycle Frames. Journal of 

Forensic Identification, 2008, 58(1): 27-32. 

(136) da-Silva L and dos-Santos PAM, Recovering Obliterated Laser Engraved 

Serial Numbers in Firearms. Forensic Science International, 2008, 179(2-3): 

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86

Examination of Glass 


Jodi Blakely Webb, M.S. 

Federal Bureau of Investigation Laboratory 

Quantico, Virginia USA 22135 

Jodi.webb@ic.fbi.gov 

87

  



2.  OVERVIEW  89  

2.1  United States National Academy of Sciences report, “Strengthening  Forensic Science in  

  the United States:  A Path Forward.”  90  

2.2  Conferences and Symposia  92  

3.  MANUFACTURING NEWS AND TRENDS  97  

4.  GLASS EXAMINATIONS  98  

5.  TRANSFER AND PERSISTENCE  102  

6.  STATISTICS  103  

7.  ACKNOWLEDGEMENTS  105  


      

88


This review covers relevant publications in the field of forensic glass analysis 

since the last report, presented at the 15 th Interpol Forensic Science Symposium 

in October 2007. Included in this report are pertinent journal publications and 

highlights from forensic meetings and symposia. 

Additional developments in the glass manufacturing industry should also be of 

interest to the forensic community. Glass manufacturing is a dynamic industry 

encompassing the research, development, and production of flat, container, 

optical, and specialty glasses. It employs a large body of researchers constantly 

developing, strengthening, enhancing, and streamlining specific processes and 

products. Significant changes and trends in research, development, and 

manufacturing that directly affect the results of forensic glass analysis, especially 

regarding product distribution, composition, and thermal history will be 

highlighted in this report. 

2. Overview 

Noteworthy work in the field of forensic glass analysis in the last three years 

includes continued work with elemental analysis, multivariate statistics, and 

interpretation. In addition to the forensic literature, articles important to forensic 

glass analysis have been published in the following journals: Analytical 

Chemistry; Analytica Chimica Acta; Applied Spectroscopy Reviews; American 

Ceramic Society Bulletin; Archaeometry; Chemometrics and Intelligent 

Laboratory Systems; Emerging Ceramics and Glass Technology; Intelligence 

Data Analysis; Journal of Analytical Atomic Spectrometry; Science; Science and 

Technology; and X-Ray Spectrometry. The American Mineralogist (1) stated that 

in response to requests from the Mineralogical Society of Great Britain and 

Ireland, The Mineral Society of America, The Mineral Association of Canada, and 

Société Française de Minéralogie et de Cristallographie, it would start publishing 

a series of papers dealing with amorphous materials such as glass, to include 

topics on properties, structure, and durability. A new manufacturing journal, the 

International Journal of Applied Glass Science, published its inaugural issue in 

March 2010 themed: “Modern Aspects of Non-Crystalline Solids.” This promises 

to be an informative and valuable addition to the arsenal of glass manufacturing 

information. 

Additional resources are available on the World Wide Web. Several websites 

offer information on product supply, manufacture and distribution that may aid the 

glass analyst. Some of these are listed here: 

The Glass Association of North America (GANA) is an association serving flat 

glass manufacturers, fabricators, and glazing contractors 

(www.glasswebsite.com). 

89

The Glass Manufacturing Industry Council (GMIC) is devoted to research on how 

to strengthen glass (www.gmic.org). 

The International Collaboration on Glass Mechanics promotes cooperation 

among scientists and engineers active in areas of glass mechanics (www.glassfracture.org). 

The National Glass Association (NGA) promotes education, workmanship, ethics, 

and safety in the architectural, automotive, and window and door glass industries. 

The NGA acts as a clearinghouse for industry information (www.glass.org). 

Additionally, the website www.glassmagazine.com promotes itself as, “the official 

magazine” of the National Glass Association, and provides subscribers glass 

industry news, trends, glass industry statistics, and supplier resource guides. 

The website www.glassonweb.com serves as a source of information on flat 

glass. 

Finally, the web service www.glassfiles.com is dedicated to collecting and 

distributing educational information on the global architectural and automotive 

glass industries. 

2.1 United States National Academy of Sciences report, “Strengthening 

Forensic Science in the United States: A Path Forward.” 

In the last three years, much of the emphasis in the forensic community has been 

on interpretation, standardization and reporting, at least partially a result of the 

United States National Academy of Sciences (NAS) report, “Strengthening 

Forensic Science in the United States: A Path Forward.” Although the report did 

not contain specific criticisms of forensic glass analysis, it did include criticisms of 

and recommendations for the trace evidence discipline in general (2). 

Nevertheless, reaction to the report reaffirms the need to improve the quality of 

all forensic science disciplines. 

The NAS report has had a serious impact on the United States forensic science 

community, specifically because of extensive and mostly negative media 

coverage. The forensic community is responding. In May, 2009 Acting Director 

of the Bureau of Alcohol, Tobacco, Firearms and Explosives of the United States 

Department of Justice Kenneth Melson addressed the US Congress. In general, 

Mr. Melson argued that the forensic community welcomed the NAS report as a 

positive contribution to the forensic community, but cautioned the public that the 

report should not undermine the use of forensics in general or offer judgments on 

cases currently in the judicial system (3). The Office of Science and Technology 

of the President of the United States has developed a subcommittee on forensic 

90

science. The goal of this committee is to establish clear, national goals and 

standards for forensic science, working in the areas of Education and Ethics; 

Standards, Practices, and Protocols; Research and Development; Accreditation 

and Certification; and Outreach. 

In addition to national efforts, individual organizations have also responded to the 

NAS report. An article by Budowle et al. (4) entitled “A Perspective on Errors, 

Bias and Interpretation in the Forensic Sciences and Direction for Continuing 

Advancement” offers a response from the United States Federal Bureau of 

Investigation (FBI). The authors attempt to clarify the issue of error, namely 

measurement error versus human error, and bias. They also provide a list of 

additional recommendations for advancing the forensic sciences, including 

developing strategic plans and initiatives for discipline-specific research needs, 

publishing research needs and validation criteria, developing national databases 

on reference materials for better assessment of the weight of the evidence, and 

defining proper language for conveying the meaning of an association. 

Moreover, the NAS report seems to have had a far-reaching influence; forensic 

scientists worldwide have begun responding to the criticisms outlined in the 

report. For example, twenty-two European Union forensic scientists met in 

August, 2009 in Wiesbaden, Germany to discuss the implications and potential 

impact of the NAS report to Europeans. An introspective summary report was 

generated in which the participants maintain that forensic science in Europe is 

“seen to be well ahead” of the United States, but still could work towards 

improvements in reporting and transparency (5). 

Much of the criticism from the NAS report addressed the overall lack of reporting 

standards. In January, 2009, the Scientific Working Group for Materials Analysis 

(SWGMAT) published an Expert Reporting Guideline (6). This document 

describes the minimum elements that should be included in a forensic report of 

examination to satisfy the requirements of the American Society of Crime 

Laboratory Directors Laboratory Accreditation Board (ASCLD/LAB)-International 

– Supplemental Requirements, ISO/IEC International Standard 17025, and 

United States Federal Rules of Evidence. It also details exactly the type of 

information the NAS would like to see in a forensic report of evidence. It should 

be noted that this guideline was published prior to the release of even the draft 

version of the NAS report (February, 2009), but was not mentioned therein, nor 

was it mentioned in the final version of the report, released in September of 2009 

(7). 

In a presentation at the 2009 American Academy of Forensic Sciences (AAFS) 

meeting, Becker discussed the accreditation of the German Bundeskriminalamt 

(BKA) to the ISO 17025 and 17020 standards (8). He advocated aspiring to 

these or similar standards. Accreditation to these standards will help insure a 

high level of quality and transparency in reporting. Many laboratories worldwide 

are adopting these rigorous standards. 

91

The NAS report will undoubtedly have other more sweeping effects than those 

that have already occurred. As forensic examiners discuss the findings of the 

NAS, many are looking critically at the underlying science of what they do (9). 

With respect to glass, Bottrell (10) provides a comprehensive report on the 

background information forensic glass examiners draw upon when interpreting 

their data. This lays the foundation for forensic glass examinations and its 

discrimination potential, and can be used by scientists and legal professionals 

alike. Data interpretation focuses on current practices of the FBI Laboratory and 

is not meant to represent interpretation practices worldwide. Additional research 

will be done in the areas of validation and data interpretation. While no results 

have yet been released, the United States National Institute for Justice (NIJ) is 

supporting an Elemental Analysis of Forensic Evidence Working Group (11) that 

is looking into appropriate statistical schemes for interpreting trace evidence 

results, including those for glass analyses. Their success could mitigate many of 

the criticisms in the NAS report. 

2.2 Conferences and Symposia 

During the last three years, many forums were held that reported on the current 

status of the glass industry and various technologies that impact forensic glass 

analysis. Additionally, meetings in the areas of glass and ceramics research and 

fractography offer information that should not be overlooked by glass analysts. 

2.2.1 American Ceramic Society (ACS) 

In 2007, the ACS published the proceedings of the Fifth Conference on the 

Fractography of Glasses and Ceramics held in Rochester, New York (USA) in 

2006 (12). Topics of interest to the forensic glass analyst include: the fracture 

process; contact damage; fracture patterns of impact resistant glasses; and 

analysis of projectile damage through bulletproof glasses. The sixth conference 

is scheduled for 2011 in Jacksonville, Florida (13). 

2.2.2 AAFS 

In the last three years, the AAFS held their annual meetings in Washington, DC 

in 2008; Denver, Colorado in 2009; and Seattle, Washington in 2010. Several 

new or novel techniques seldom used in forensics were described for the 

elemental analysis of glass. The NAS report, its fallout, and subsequent 

recommendations were discussed extensively at the 2010 meeting. 

Sampson et al. (14) presented a report about the potential use of ion beam 

analysis for discriminating glass fragments. The authors argue the advantages 

of ion beam analysis as being non-destructive, little sample preparation, and 

92

higher sensitivity than x-ray fluorescence. Scarcity in forensic laboratories, 

however, may hinder its general acceptance. 

Van Es et al. (15) discussed a likelihood ratio approach using multivariate glass 

data analyzed via laser ablation-inductively coupled plasma-mass spectrometry 

(LA-ICP-MS). Van Es also presented a comparison between the Bayesian 

approach and a “fall-off-the-cliff” approach using confidence intervals. Both type 

I and type II errors were demonstrably lower when calculating a likelihood ratio. 

Martinez et al. (16) demonstrated the potential for yet another technique for the 

elemental analysis of glass, using infrared microprobe analysis with diamond 

attenuated total reflection. Use of this technique will require the development of 

specialized spectral databases. 

Almirall, Perez and Barnett (17) showed that Laser Induced Breakdown 

Spectroscopy (LIBS) can readily discriminate vehicle versus container glass. 

This continues their previous work demonstrating that LIBS has comparable 

results to other, more established elemental analysis techniques for glass. 

Gardner and Broach (18) also presented a study on the reliability and 

discriminating capabilities of LIBS. And Cahoon and Almirall (19) continued to 

advocate the reliability and cost-efficiency of LIBS. 

2.2.3 Asian Forensic Sciences Network (AFSN) 

A new group conceived in 2008, the AFSN, held its inaugural meeting in 2009. 

This network will serve as the collective representation for the forensic science 

community in Asia. Currently there is a Trace Evidence Working Group with 

about 8 members (20). They have yet to produce any publications regarding 

glass. 

2.2.4 Australian and New Zealand Forensic Science Society Inc. (ANZFSS) 

The ANZFSS held its 19 th International Symposium in Melbourne, Australia in 

October, 2008. The symposium was titled “Domestic Crime to International 

Terror.” This organization holds symposia every two years. The 20 th 

International Symposium will be held in Sydney, Australia in September, 2010 

(21). 

Wong et al. (22) produced a study on the effects different types of projectiles 

have on glass backscatter. Results showed that an increase in the velocity of a 

projectile had an exponential increase in the total number of backscattered 

fragments. While this seems intuitive, these results contradict previous studies 

(23, 24). Additionally, the authors designed a statistical model for estimating the 

number of expected fragments given the velocity, hardness and profile of a 

projectile. 

93

It is empirically understood by most glass analysts that the distribution of 

refractive index (RI) values of recovered glass fragments is commonly wider than 

those of the control glass values. Newton (25) discussed possible explanations 

for this phenomenon. 

Smith et al. presented case studies touting the value of the analysis scheme of 

RI and LA-ICP-MS for forensic glass comparisons (26). 

2.2.5 European Ceramics Society and European Structural Integrity Society 

The European Ceramics Society and European Structural Integrity Society 

organized a conference in September, 2008 at the Institute of Materials 

Research, Stara Lesna, Slovakia, entitled the 3 rd International Conference 

“Fractography of Advanced Ceramics.” Similar to its American counterpart, this 

conference focused on damage mechanisms and fracture processes. The next 

such meeting is scheduled for 2012 (27). 

2.2.6 European Network of Forensic Science Institutes (ENFSI) 

The European Academy of Forensic Sciences (EAFS) of the ENFSI meets every 

three years and held their fifth meeting in Glasgow, Scotland in 2009. There, 

Zadora (28, 29) presented his continuing work on forensic glass interpretation. 

The European Paint and Glass (EPG) Working Group of the ENFSI meets 

annually. Their three meetings were held in Lisbon, Portugal in 2007; Delft, The 

Netherlands in 2008; and St. Gallen, Switzerland in 2009. The EPG offers yearly 

proficiency tests and collaborative exercises in glass. Best practice guidelines 

are drafted and available to members on their website. Annual meetings provide 

an abundance of research presentations and interesting case studies. 

Weis (30) presented several case studies touting the discriminating power of LA- 

ICP-MS versus the Foster and Freeman, Ltd. Glass Refractive Index Measuring 

(GRIM) system. In every case but one, LA-ICP-MS was able to distinguish 

glasses that were indistinguishable by RI. In that one instance, RI was able to 

distinguish between two glasses that were chemically indistinguishable. Thus, 

Weis concluded that the combination of LA-ICP-MS and RI determination leads 

to excellent discrimination and classification. 

In an interesting case where glass fragments were too small to see and recover 

using traditional microscopic methods, Sjåstad (31) used scanning electron 

microscopy with energy dispersive x-ray spectroscopy (SEM-EDS) to detect and 

analyze glass fragments found on stolen banknotes. Glass fragments less than 

0.5mm were found on these banknotes and were consistent with broken glass 

from the scene of a large, violent bank robbery. 

A poster presented by Lauper (32) compared ramp rates for RI analysis using the 

GRIM system and found that 4˚/min was the optimal rate. The authors also 

94

caution about the environmental sensitivity, namely fluctuating precision with 

temperature, humidity, etc., of the GRIM. The authors correctly indicate that if all 

fragments from a given case are analyzed on the same day users may be able to 

avoid problems associated with extensive instrument drift. 

Krieghoff (33) discussed his experiences with the new LUCIA RI: Refractive 

Index Measurement System as compared to the GRIM system. 

Zadora (34) describes the application of Bayesian nets to continuous data. A 

Bayesian net is a graphical model used to show whether two sets of data, RI of 

glass in this case, could share a similar source. 

In Delft and St. Gallen, Zadora (35, 36) demonstrated software (calcuLatoR) 

used to calculate a likelihood ratio using univariate (RI) data. While this offers 

the glass analyst a simple solution to the difficulty of calculating a likelihood ratio, 

analysts should not use this program without a thorough understanding of the 

variables used and underlying assumptions of the method. 

Zadora and Neocleous (37) compared various methods of databasing univariate 

data for within-object variability and their effects on likelihood ratio determinations. 

Most forensic laboratories use in-house databases to test within- and betweenobject 

variability. These are generally constructed using a relatively small 

sample set, usually from case work, to represent a larger object or objects. 

While databases of this kind may not necessarily represent the exact population 

of broken glass for a given case, the authors concluded that databases 

generated in this fashion are appropriate to use for likelihood ratio calculations. 

Dätwyler, Fehr and Ivo (38) used total reflection x-ray fluorescence (TXRF) for 

elemental analysis of glass. While this technique allows for detection limits in the 

parts per billion (ppb) range for many elements, the precision is strongly 

dependent on appropriate sample preparation. Samples prepared in 

suspensions varied less than powdered glass samples placed directly on a 

sample holder. 

Additionally, the EPG commenced publishing an annual newsletter. In the 2009 

edition, Becker (39) edited and published “The forensic analysis of float-glass 

characterization of glasses from international sources.” Originally presented by 

Wilfried Stoecklein at the 1997 Trace Evidence Symposium in San Antonio, 

Texas (USA), the proceedings from that conference, while often discussed, were 

never published. The work carried out by the BKA in forensic glass analysis in 

the 1990’s is reported therein. 

2.2.6 NIJ Symposia 

In August 2007, the US NIJ and the FBI Laboratory sponsored a symposium 

entitled “Making the Case with Trace Evidence” in Clearwater, Florida (USA), an 

95

internationally attended event, with representatives from 17 countries and 

approximately 350 attendees (40). The conference was important to the forensic 

science community because the last symposium of its kind focusing on trace 

evidence was in San Antonio, Texas (USA), in 1997. In the glass field, several 

papers were presented emphasizing the value of elemental analysis (41); 

specifically LA-ICP-MS (42, 43). Additionally, many workshops were given, 

offering training and fostering discussion. In particular, the workshop entitled 

Advanced Elemental Analysis of Trace Evidence Examinations, dealt with topics 

in forensic glass analysis. This event was so successful and well-received that it 

is slated to be held biannually. 

The second Trace Evidence Symposium was held in 2009 (44). Where the 2007 

symposium agenda was fairly generalized, the 2009 Trace Evidence Symposium 

offered more workshops and specialized topics, supplying analysts with further 

training opportunities. Workshops on glass included The Use of Multivariate 

Statistics in Trace Evidence Investigations, Advanced Elemental Analysis, 

Physical Match Theory, Standardized Test Methods and Insuring Quality in the 

Trace Laboratory, and Report Writing. This offered yet another opportunity to 

focus on responding to the NAS report, both refuting some of the findings and 

challenging forensic scientists to improve. 

2.2.7 SWGMAT 

The SWGMAT met in conjunction with the 2007 Trace Evidence Symposium and 

separately in Fredericksburg, Virginia (USA) in 2008 and 2010. The SWGMAT is 

very much focused on producing best practice guidelines for trace evidence 

analysts. Once again, interpretation, standardization and reporting were heavily 

debated. Some case studies were presented. 

A presentation by Eyring (45) showed the use of a microspectrophotometer for 

the objective determination of the color of glass fragments. The size required for 

optimal analysis is larger than most recovered glass samples. Therefore, use of 

this technique is not likely to be widespread. 

As mentioned previously, the SWGMAT published the “Expert Reporting 

Guideline” governing how glass analysts should write reports (46). Additionally, it 

was reported (47) that the Glass Sub-Group of the SWGMAT is drafting guidance 

for interpretation of results, and meeting admissibility requirements in courts. 

The 2010 SWGMAT meeting concentrated primarily on building and defining 

significance scales for trace evidence interpretation, but no guidelines on the 

subject have been issued to date. A new Interpretation Sub-Group was formed 

that will strive for some consistency across disciplines. 

A transfer and persistence study by Green and Sheedy (48) found that the 

uppers of the shoes have the greatest potential for particle recovery as they 

96

etained the largest number of glass fragments. The authors advocate collecting 

debris from the uppers separately from the bottoms of shoes. In every test, glass 

fragments were recovered from people near the breaking glass who were not 

involved with the breaking. Even though the same tool was used for each 

experiment and the breaking strength was roughly consistent throughout, the 

amount of recovered glass was different for each experiment. 

3. Manufacturing News and Trends 

Glass continues to be a large, dynamic enterprise which boasts ongoing 

research and development for standard use glass (e.g. automotive, architectural, 

container) and developing uses (e.g. solar energy, nuclear waste disposition). 

Development continues in the areas of coatings, glass strength, and thermal 

efficiency. Rising fuel costs are causing an increase in the cost of raw materials, 

resulting in an increase (“single digit percentage”) in glass prices, including 

vehicle glass (49). The high cost of fuel coupled with legislation limiting carbon 

emissions may drive the glass industry to different batch compositions. For 

example, the use of lithium-containing feldspars is being studied because they 

have the potential to reduce the need for limestone while lowering melting 

temperatures and energy costs (50). 

Although North America remains a major producer of raw materials, glassmaking 

in North America is decreasing. China has become the dominant producer of flat 

glass (51). China is also the world’s largest producer of fiberglass, producing 

37% of the total global output in 2008 and exporting 1.085 million tons. In the 

same year, China produced 50 million square meters of Low-E glass, and 

imported 47 million square meters more (52). This has implications in the 

interpretation of glass; in particular, frequency of occurrence statistics. For 

example, the frequency of a particular RI value will likely change and narrow to 

reflect a larger percentage of glass being built in the same location from similar 

raw materials. Conversely, in some countries with dire economic conditions it is 

still more economical for glass manufacturers to use raw materials from local 

deposits and produce glass for limited distribution. Local deposits, however, may 

contain more impurities that can compromise the quality of the glass, but may 

impart a more unique chemical signature and physical appearance (53, 54). 

Nevertheless, the glass manufacturing industry is undergoing increasing 

consolidation into several large companies, with smaller companies being bought 

out. Belron, US, the parent company of Safelite has purchased Cindy Rowe and 

Diamond Triumph (55). PPG closed several plants in North America and sold its 

automotive glass and services business to Kohlberg & Company, LLC. PPG’s 

remaining glass operations were re-branded “Pittsburgh Glass Works,” or PGW. 

PGW will focus on specialty glass for the solar energy market. PGW is heavily 

investing in upgrading and improving their remaining plants (56). 

97

Several new glass products were introduced in the last three years. In 2008, 

Saint-Gobain, in partnership with the Saazs Institute, began producing Planilum, 

a light emitting glass. Planilum consists of four layers of tin-doped indium oxide 

conducting glass, containing an ionized plasma gas (57). In 2009, Vitro 

Cristalglass began producing a “self-cleaning” glass called “Ecopure.” This glass 

is coated with a hydrophobic coating that boosts visibility during rain, and 

destroys organic contaminants on its surface. The make-up of this coating is 

proprietary, and was not reported (58). Also, Owens Corning is increasing by 

40% the production of “Advanex” - a composite material using bundles of 

continuous filament glass fiber combined into a single strand (59). 

The glass manufacturing industry continues to invest significant resources into 

basic research. For example, Ponsich et al. model glass manufacturing with an 

emphasis on the description of the furnaces (60). And Hedges et al. offer 

numerical evidence for the existence of the hypothetical and controversial glass 

phase transition (61). 

4. Glass Examinations 

4.1 Fractography 

Failure analysis of brittle materials continues to be a widely studied field. 

Although abundant research exists on fractography of glass, new literature 

continues to be published. Forensic glass analysts should be familiar with 

fracture mechanics when reconstructing broken glass objects. 

The Handbook of Glass Fractography by R.C. Ropp (62) has well-written 

information about glass making, history, composition, processes, strength, and 

end use. The author has twenty years of experience as an expert witness, 

mostly in civil litigation where the expert is not necessarily constrained by 

accreditation guidelines. Nevertheless, Ropp imparts his knowledge and 

perspectives to the reader through a number of interesting cases. 

Overend, De Gactano and Haldimann (63) provide an overview of the mechanics 

of glass fracture under loading stresses that is intended for use by the consulting 

engineer who conducts forensic investigations of glass failure. Increasing use of 

glass panels in non-traditional areas such as floors, roofs and staircases where 

large loads are common make this important to civil as well as criminal litigation. 

Natanail and Duse (64) published a short study on the damage, hardness, and 

physical defects of float glass in the Annals of Danube Adria Association for 

Automation and Manufacturing (DAAAM). The authors test the strength of float 

glass by simulating defects that may be caused during- or post-manufacturing. 

98

4.2 Optical Measurements 

There have been no significant technical advances in the instrumentation used 

for refractive index determination. However, LUCIA Forensic has developed a 

new system for automatically determining the refractive index of small glass 

particles: the LUCIA RI: Refractive Index Measurement System. LUCIA Forensic 

hopes to compete with the Foster and Freeman, Ltd. Glass Refractive Index 

Measuring System (GRIM). The LUCIA RI: Refractive Index Measurement 

System is comparable to GRIM and represents no breakthroughs in technology, 

but is lower in cost and has the ability to measure more glass edges 

simultaneously. Currently this system is only available in Europe. 

Koons and Garvin (in press) compare the discrimination potential of eight 

commonly used match criterion for RI comparison of flat glass fragments. RI 

determinations were performed with a GRIM, applying the “bootstrapping” 

method for sample selection and statistical evaluations (65). This work was 

presented at the 2009 AAFS and EPG meetings (66, 67). Glass analysts will 

have a better understanding of the error potential of their chosen criteria as this 

study determines type I error rates for each different match criteria, using 

different numbers of control measurements. Adequate characterization of control 

glasses (10 measurements or more) gives low type I error rates. Once again, 

these data show that RI is generally not normally distributed and the analyst is 

cautioned in choosing a criterion with an assumption of normality. More work 

addressing type I error rates for elemental data and type II error rates for RI and 

elemental data is planned. Work of this kind directly addresses admissibility 

issues and criticisms in the NAS report regarding the lack of known error rates in 

many forensic disciplines. 

Newton and Buckleton (68) show that glass fragment edge morphology affects 

the results of RI measurements using the GRIM system. Edge count values 

decrease with poor edge morphology. As edge count values decrease, the 

variation in RI increases. Experienced users of the GRIM systems are familiar 

with this occurrence and often instigate an arbitrary cutoff value of 10 as 

recommended by Foster and Freeman, Ltd in the original GRIM manual (69). 

Newton and Buckleton opine that poor edge morphology may be one explanation 

for the wider distribution of RI values for recovered fragments (70). They 

suggest a minimum edge count value of 30. Although this higher edge count 

value may limit the number of recovered fragments acceptable for measurement, 

the precision of the resulting RI data will likely increase. 

Brown et al. attempt to classify mineral wool fibers and glass fibers from World 

Trade Center dust, using classic methods of RI and SEM-EDS (71). The SEM- 

EDS data of a single mineral wool fiber was fairly consistent, whereas the 

variation in elemental composition among fibers within a single product was 

widely variable. As with other work on mineral wool, some classification is 

possible, but several manufacturers’ products overlap using these techniques. 

99

Another paper revisiting older topics is by Pawluk-Kołc, Zięba-Palus and 

Parczewski (72) who examined the classification and discrimination potential of 

annealing and change in RI. 

4.3 Elemental Composition 

Significant work continues on the elemental analysis of glass for comparison and 

classification purposes. LA-ICP-MS continues to be the recommended method 

of elemental analysis with many advocating the combination of RI and LA-ICP- 

MS as the most discriminating analysis scheme to date (73, 74). The method of 

LIBS is touted as a cheaper, non-destructive alternative with multi-element 

capability as has been documented in numerous studies. Two newer and novel 

techniques, time of flight-secondary ion mass spectrometry (TOF-SIMS) and 

synchrotron radiation x-ray fluorescence spectroscopy (SR-XRF), have shown 

good discrimination capabilities, but with the high associated cost and limited 

availability of these techniques it is doubtful that either will become the new 

standard. 

May and Watling (75) studied the RI and elemental variation (by LA-ICP-MS) of 

glass containers from a single manufacturing plant, produced over time. The 

discrimination capability of the analytical techniques and interpretation protocols 

were established. They confirm (as previous studies have also) that the variation 

of RI between and within bottles renders this examination technique of limited 

discrimination potential when used by itself. Also, the statistical method chosen 

for the evaluation of RI is dependent upon end use, i.e., whether RI values are 

used as a screen prior to elemental analysis or as the sole analytical technique 

used to discriminate fragments. While it is expected that many bottles will be 

chemically indistinguishable, LA-ICP-MS data showed that the elemental 

assemblage within a single bottle was uniform and specific enough to distinguish 

between most glass bottles packaged within a single crate. Even though it is not 

possible to discriminate between all bottles, this method provides significant 

improvement over RI alone. 

Van Elteren, Tennent and Šelih (76) study the potential of using LA-ICP-MS to 

analyze ancient/historic glasses for archaeometric investigations. Unlike modern 

glass, these samples are inherently inhomogeneous with widely ranging 

compositions. For this reason, the normal method of using matrix-matched 

external standards is not appropriate. To overcome some of these issues, the 

authors use a raster method for sample introduction as opposed to crater 

sampling, and a mathematically formulated “sum normalization calibration 

technique”. This technique shows promise for its application to the analysis of 

unknown or inhomogeneous glass types in forensics, but further study would be 

required. 

100

Additional scientists working in the field of Archaeology and Art History use SEM- 

EDS and LA-ICP-MS to help determine the origin and differentiation of ancient 

glass. Dussubieux et al. (77) and van der Linden et al. (78) validate the 

usefulness of quantitative trace element analysis in the study of glass. Work of 

this kind is analogous to forensic science in that it is also reconstructing past 

events and speaks to the general acceptability of the techniques. 

Scientists working in the earth sciences also contribute to the validation of LA- 

ICP-MS. Similar to the work of forensic scientists, Gagnon et al. (79) discuss the 

use of an internal standard for quantitative analysis of unknown silicate materials. 

Naes et al. (80) compare the well-tested methods of LA-ICP-MS and micro X-ray 

fluorescence spectroscopy (µXRF) to the newer LIBS method for the analysis of 

automotive glass. LA-ICP-MS is still superior in accuracy, precision, sensitivity, 

and discriminating potential. Regardless, with the high cost and relative 

complexity of operation of LA-ICP-MS, LIBS may be a viable alternative. The 

authors developed an optimal analytical scheme designed to eliminate Type I 

errors and reduce Type II errors. 

Bridge et al. (81) conducted another assessment of LIBS versus LA-ICP-MS 

using different glass types to compare their discrimination potentials. The 

samples were also examined by RI. Glass samples compared included 

automotive side and rear windows, headlamps, and mirrors; and brown glass 

bottles. As determined from other studies (82), the combination of LA-ICP-MS 

and RI gives the best discrimination (98.8%) over the scheme of LIBS plus RI 

(87.2%). The side-mirrors were particularly problematic when analyzed by LIBS. 

Roderiguez-Celis et al. (83) explore the potential of LIBS for identifying glass 

fragments according to their spectral “fingerprints,” study long-term reproducibility, 

and report optimal sampling conditions. LIBS has advantages of little sample 

prep, good throughput, and simultaneous, multielement analysis. While LIBS 

has low absolute detection limits, LIBS is hampered by poor precision, poor 

relative detection limits, and matrix and spectral interferences. Rather than 

obtaining detailed chemical composition data, the authors seek to establish that 

each glass source has a unique spectral “fingerprint” that may be obtained using 

statistical correlation models. An unknown glass is identified by correlating its 

spectra to a database. The spectra are then compared against each other to 

determine if they could have originated from the same source. The authors seek 

to demonstrate that LIBS may be a viable and more cost effective alternative to 

LA-ICP-MS. 

Barnett, Cahoon and Almirall (84) continue the work by Florida International 

University on LIBS and work toward establishing best possible instrumental 

conditions and validation. They characterize the influence of the irradiation 

wavelength and determine the best lens-to-sample distance for each wavelength 

used in order to optimize precision. 

101

Columbaros et al. (85) use TOF-SIMS to evaluate the elemental variation across 

and through a sheet of glass, particularly the non-float surface. TOF-SIMS offers 

the potential for analyzing glass fragments as small as tens of microns across. 

As with RI, there is measurable variation with TOF-SIMS analysis between the 

surface and bulk. This supports the practice of avoiding surface fragments when 

conducting RI analyses. Results show that evaluating the non-float surface is not 

straightforward, and certainly not representative of the bulk. The authors 

recommend that when techniques such as SIMS are used, which operate to 

depths measured in nanometers, extra caution should be used to avoid type I 

errors and suggest eroding the surface to expose the bulk. 

Nakanishi et al. (86) explore the use of synchrotron radiation high-energy X-ray 

fluorescence (SR-XRF) spectrometry for the nondestructive discrimination of 

glass. SR-XRF offers the advantage of extremely low detection limits for some 

elements, but those values had not yet been quantified. This study was 

undertaken to discover the lower limits of detection for elements found to be 

discriminating for glass. Detection at the pictogram (pg) level was possible for 

Ba, Ce and Sm, and at the 10-pg level for Sr, Zr, Sn and Hf. The elements Ca, 

Mn, Fe and Sn were not used as they had unacceptably high relative standard 

deviations (greater than 10%). In general, however, satisfactory discrimination 

was achieved and the technique was determined to be effective for a 

nondestructive analysis of trace amounts of heavy elements contained in a 

minute sample. Currently, this instrumentation is not readily available to forensic 

laboratories. 

A feasibility study by Bell, Nawrocki and Morris (87) uses cathodoluminescence 

(CL) spectroscopy to discriminate between different glass sources. Using this 

method in conjunction with RI analysis, all window and container glasses of the 

sample set were differentiated as were six out of ten headlamps. Inter- and intrasample 

variation of several NIST reference glass standards was determined. 

The reference glasses were readily distinguishable by this method, but some 

intra-sample variation was observed. Although non-destructive, fairly 

inexpensive and easy to operate, CL is hampered by lack of standards and 

databases, and relatively low spectral resolution. Also, the relatively limited 

forensic applications of CL make it unlikely that CL will become standard in 

forensic laboratories. This work was also presented by Barron and Bell (88) at 

the 2008 AAFS meeting in Washington, D.C. 

5. Transfer and Persistence 

There were relatively few published studies on the transfer and persistence of 

glass during the reporting period. Those that were published were targeted to a 

particular environment in hopes of establishing relative populations for likelihood 

ratio calculations. In general, these studies reaffirmed other recent studies 

indicating that most of the clothing in the general population is free from glass 

102

fragments unless having been in recent contact with broken glass. Analysts 

calculating a likelihood ratio should use the most appropriate data from a relevant 

population, for their particular case. These newer studies should supplant older 

studies, such as the Pearson, May and Dabbs (89) study, which is often used for 

calculating a likelihood ratio, but out of date. 

Brożek-Mucha (90) examined the persistence of glass on clothing from 35 

different experiments, using various types of multiple-glazed windows. The 

number of fragments recovered from the clothing of the breaker and 

accompanying people depended on the distance from the window, time between 

event and recovery, the retentive properties of the clothing, and the kind of glass 

broken. Background levels were also studied. Whilst targeted specifically for 

likelihood ratio calculations in Poland, this study confirms the findings of other 

recent transfer and persistence studies: most clothing in the general population is 

free of glass. 

Nic Daeid, McColl and Ballany (91) determined background levels of glass on 

fleece jackets commonly worn in Scotland. The sample set was selected from 

individuals who work in law enforcement, either in a forensic laboratory or 

regularly processing crime scenes. Contrary to the aforementioned Brożek- 

Mucha study, about 14% of the jackets in this sample set had glass on them. RI 

values from the recovered fragments indicated that the glass was from multiple 

sources. Information about the proximity of the individuals to recently broken 

glass and laundering information about the garments was also reported. These 

data may account for the relatively high percentage of recovered glass. 

6. Statistics 

Statistical evaluations of glass data were numerous in the reporting period, with 

the Bayesian method being the most published approach for determining the 

weight of the evidence. Zadora in particular was prolific in his publications and 

presentations advocating the use of Bayesian statistics for forensic glass 

analysis. The previously mentioned Koons and Garvin (92) publication takes a 

frequentist approach. 

Zadora (93) assessed the efficiency of three methods – the likelihood ratio, 

support vector machines, and naïve Bayes classifiers – for the classification of 

glass objects using RI values and SEM-EDS data. RI values for both pre- and 

post-laboratory annealing were shown to have better discriminating capability 

than pre-annealed RI values alone. Although the likelihood ratio model was not 

the best performer, Zadora still advocated its use as being easier to interpret and 

having fewer associated assumptions. The classification of automotive versus 

architectural glasses is particularly problematic as the analytical scheme used 

does not discriminate enough for accurate categorization. 

103

Continuing the work of Curran et al. (94), Campbell and Curran (95) introduce a 

“permutation testing approach” to the interpretation of elemental analysis data 

from glass evidence. In previous work, the authors suggested using Hotelling’s 

T 2 to evaluate multivariate data that may be used in a Bayesian calculation. 

Criticisms of this approach are addressed; namely, the assumption of normality, 

and the relatively large sample size not necessarily available to the forensic 

scientist. The authors had some success removing these constraints using a 

shrinkage estimator. The reader is directed to regard this as a “pre-screening” 

tool and not for interpretation for the courts. 

Zadora and Neocleous (96) and Zadora (97) employ a Bayesian approach for 

classification of glass fragments into use-type categories using RI and SEM-EDS 

data. Support vector machines, naïve Bayes classifiers, and Bayesian networks 

were again assessed as well as the performance of the likelihood ratio models 

when considering between- and within-object variability. A model that takes both 

into consideration performs better than a model using between-object variability 

only. The accuracy of the classification is highly dependent upon glass type, but 

in general, the combination of SEM-EDS and change in RI values with annealing 

increased anywhere from 3% to greater than 10% over SEM-EDS and preannealing 

RI data alone. Caution should be taken when using this type of data 

alone for classification purposes as the values for several glass types (e.g. 

architectural and automotive) tend to overlap. Classification based on preannealed 

RI values alone should not be attempted. Likelihood ratios from these 

data are usually close to one, giving very limited support to either the 

prosecution’s or defense’s hypothesis. 

Jensen and Shen (98) explore the use of statistical feature selection as an aid to 

Bayesian analysis of glass. In fact, they aim to remove the need for expert 

knowledge in feature selection and advocate univariate data analysis for 

likelihood ratio calculations. While the use of automated feature techniques may 

be of use in evidence evaluation, it is unadvisable to rely on statistical modeling 

of a likelihood ratio without sufficient knowledge of the underlying analytical data. 

Zadora and Ramos (99) study the influence of adequate population databases 

for likelihood ratio calculations. Databases assessed are based on RI and SEM- 

EDS data. This study was also presented at the 2009 EFS meeting in Glasgow, 

Scotland (100) and the 15 th Annual meeting of the EPG in St. Gallen, Switzerland 

(101). 

This concludes the Glass Report for 2010. Future papers and presentations will 

be summarized in the next report. 

104

7. Acknowledgements 

The assistance of Maureen Bottrell and David Korejwo in reviewing the literature 

and editing this manuscript is gratefully acknowledged. 

Names of commercial manufacturers are provided for identification purposes only, 

and inclusion does not imply endorsement of the manufacturer, or its products or 

services by the FBI. The views expressed are those of the author's and do not 

necessarily reflect the official policy or position of the FBI or the U.S. Government. 

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48. Green DA, Sheedy A. The Case of the Questionable Glass and the 

Change of Custody. Presented at the Scientific Working Group for 

Materials Analysis (SWGMAT) Annual Meeting. Fredericksburg, VA 2010. 

49. News and Trends. American Ceramics Society Bulletin 2008 August; 

87(8):6. 

50. Merivale C. The Batch Solution Opportunity. The Glass Researcher 2008; 

17(4). In American Ceramics Society Bulletin 2008; 87(11):37-42. 

51. China Dominates World Float Glass Market. American Ceramics Society 

Bulletin 2008 August; 87(8):7. 

52. Wray P. China: Ceramics World Continues Eastward Shift. American 

Ceramics Society Bulletin 2008; 87(10):26-30. 

53. K.V. Goncharov. Use of Off-Grade Initial Materials in Glass Production. 

Glass and Ceramics 2007; 64(5-6):167-168. 

54. K.V. Goncharov. Use of unconventional raw materials for producing sheet 

glass by vertical debitense drawing. Glass and Ceramics 2007; 64(11- 

12):395-396. 

55. News and Trends. American Ceramics Society Bulletin 2008; 87(10):10. 

56. American Ceramics Society Bulletin 2008; 87(11):7. 

57. Research Briefs: Light Emitting Glass – the Next Big Thing. American 

Ceramics Society Bulletin 2008; 87(11):21. 

58. Virtro launches self-cleaning glass. American Ceramics Society Bulletin 

2009; 88(2):3. 

59. Glass Fiber Plant Expands in Amarillo. American Ceramics Society Bulletin 

2008; 87(8):6. 

60. Ponsich A, Azzaro-Pantel C, Domenech S, Pibouleau L, Pigeonneau F. A 

Systematic Approach for Glass Manufacturing Process Modeling. Chemical 

Engineering and Processing: Process Intensification 2009; 48:1310-1320. 

61. Hedges LO, Jack RL, Garrahan JP, Chandler D. Dynamic Order-Disorder 

in Atomistic Models of Structural Glass Formers. Science 2009 March; 

323:1309-1313. 

62. Ropp RC (editor). Handbook of Glass Fractography. Authorhouse, 

Bloomington, IN. 2008 pp 630. 

63. Overend M, De Gaetano S, Haldimann M. Diagnostic Interpretation of 

Glass Failure. Science and Technology 2007; 2:151-158. 

64. Natanail R, Duse D. Investigation on Float Glass for Automobile 

Windshields. Annals of DAAAM & Proceedings 2009; 20:605-606. 

65. Koons RD, Garvin E. Significance of Match Criteria for Refractive Index 

Comparison of Glass Fragments. Journal of Forensic Sciences, in press. 

66. Koons RD. Significance of Match Criteria for Refractive Index Comparison 

of Glass Fragments. Proceedings of the American Academy of Forensic 

Sciences 61 st Annual Scientific Meeting. Denver, CO 2009 pp 131-132. 

67. Webb JB. Significance of Match Criteria for Refractive Index Comparison of 

Glass Fragments. Proceedings of the European Paint and Glass Working 

Group of the European Network of Forensic Science Institutes Annual 

Scientific Meeting. St. Gallen, Switzerland 2009. 

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68. Newton AWN, Buckleton JS. An Investigation into the Relationship 

Between Edge Counts and the Variability of the Refractive Index of Glass: 

Part I: Edge Morphology. Forensic Science International 2008; 177:24-31. 

69. GRIM: An Automatic System for Glass Refractive Index Measurement. 

Foster and Freeman, Ltd. 1987. Instrument Manual. 

70. Newton 2008. 

71. Brown RS, Boltin WR, Bandli BR, Millette JR. Light and Electron 

Microscopy of Mineral Wool Fibers. Microscope 2007; 55(1):37-44. 

72. Pawluk-Kołc M, Zięba-Palus J, Parczewski A. The Effect of Re-Annealing 

on the Distribution of Refractive Index in a Windscreen and a Windowpane: 

Classification of Glass Samples. Forensic Science International 2008; 

174:222-228. 

73. Weis 2007. 

74. Smith et al. 2008. 

75. May CD, Watling RJ. A Comparison of the Use of Refractive Index (RI) and 

Laser-Ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICP-MS) 

For the Provenance Establishment of Glass Bottles. Forensic Science 

Medical Pathology 2009; 5:66-76. 

76. Van Elteren JT, Tennent NH, Šelih VS. Multi-Element Quantification of 

Ancient/Historic Glasses by Laser Ablation Inductively Coupled Plasma 

Mass Spectrometry Using Sum Normalization Calibration. Analytica 

Chimica Acta 2009; 644:1-9. 

77. Dussubieux L, Kusimba CM, Gogte V, Kusimba SB, Gratuze B, Oka R. The 

Trading Of Ancient Glass Beads: New Analytical Data From South Asian 

And East African Soda-Alumina Glass Beads. Archaeometry 2008; 50:797- 

821. 

78. Van der Linden V, Cosyns P, Schalm O, Cagno S, Nys K, Janssens K, 

Nowak A, Wagner B, Bulska E. Deeply Coloured And Black Glass In The 

Northern Provinces Of The Roman Empire: Differences And Similarities In 

Chemical Composition Before And After AD 150. Archaeometry 2009; 

51:822-844. 

79. Gagnon JE, Fryer BJ, Samson IM, Williams-Jones AE. Quantitative 

Analysis of Silicate Certified Reference Materials by LA-ICPMS With and 

Without an Internal Standard. Journal of Analytical Atomic Spectoscopy 

2008; 23:1529-1537. 

80. Naes BE, Umpierrez S, Ryland S, Barnett C, Almirall JR. A Comparison of 

Laser Ablation Inductively Coupled Plasma Mass Spectrometry, Micro X- 

Ray Fluorescence Spectroscopy, and Laser Induced Breakdown 

Spectroscopy for the Discrimination of Automotive Glass. Spectrochimica 

Acta Part B 2008; 63:1145-1150. 

81. Bridge CM, Powell J, Steele KL, Sigman ME. Forensic Comparative Glass 

Analysis by Laser-Induced Breakdown Spectroscopy. Spectrochimica Acta 

Part B 2007; 62:1419-1425. 

82. Weis 2007. 

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83. Rodriguez-Celis EM, Gornushkin IB, Heitmann UM, Almirall JR, Smith BW, 

Winefordner JD, Omenetto N. Laser-Induced Breakdown Spectroscopy as 

a Tool for Discrimination of Glass for Forensic Applications. Analytical & 

Bioanalytical Chemistry 2008; 391:1961-1968. 

84. Barnett C, Cahoon E, Almirall JR. Wavelength Dependence on the 

Elemental Analysis of Glass by Laser Induced Breakdown Spectroscopy. 

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85. Coumbaros J, Denman J, Kirkbride KP, Walker GS, Skinner W. An 

Investigation into the Spatial Elemental Distribution within a Pane of Glass 

by Time of Flight Secondary Ion Mass Spectrometry. Journal of Forensic 

Sciences 2008; 53:312-320. 

86. Nakanishi T, et al. Lower Limits of Detection of Synchrotron Radiation 

High-Energy X-Ray Fluorescence Spectrometry and Its Possibility for the 

Forensic Application for Discrimination of Glass Fragments. Science Direct 

2008; 175:227-234. 

87. Bell SC, Nawrocki HD, Morris KB. Forensic Discrimination of Glass Using 

Cathodoluminescence and CIE LAB Color Coordinates: A Feasibility Study. 

Forensic Science International 2009; 189:93-99. 

88. Barron H, Bell SC. Discrimination of Glass by Cathodoluminescence, Color 

Analysis, and Chemometrics. Proceedings of the American Academy of 

Forensic Sciences 60 th Annual Scientific Meeting. Washington, DC 2008 pp 

47. 

89. Pearson EF, May RW, Dabbs MGD. Glass and Paint Fragments Found in 

Men’s Outer Clothing—Report of a Survey. Journal of Forensic Sciences 

1971; 16:283–300. 

90. Brożek-Mucha Z. X-ray Microanalysis of Glass for Forensic Purposes – 

Research on the Persistence of Glass Fragments on Clothing. X-Ray 

Spectrometry 2009; 38:58-67. 

91. Nic Daéid N, McColl D, Ballany J. The Level of Random Background Glass 

Recovered from Fleece Jackets of Individuals Who Worked in Law 

Enforcement or Related Professions. Forensic Science International 2009; 

191:19-23. 

92. Koons RD, Garvin E In press. 

93. Zadora G. Classification of Glass Fragments Based on Elemental 

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54(1):49-59. 

94. Curran JM, Triggs CM, Almirall JR, Buckleton JS, Walsh KAJ. The 

Interpretation of Elemental Composition Measurements from Forensic 

Glass Evidence. Science and Justice 1997; 37:241-244. 

95. Campbell GP, Curran JM. The Interpretation of Elemental Composition 

Measurements from Forensic Glass Evidence III. Science and Justice 2009; 

49:2-7. 

96. Zadora G, Neocleous T. Likelihood Ratio Model for Classification of 

Forensic Evidence. Analytica Chimica Acta 2009; 642:266-278. 

111

97. Zadora G. Evaluation of Evidence Value of Glass Fragments by Likelihood 

Ratio and Bayesian Network Approaches. Analytica Chimica Acta 2009; 

642:279-290. 

98. Jensen R, Shen Q. Feature Selection for Aiding Glass Forensic Evidence 

Analysis. Intelligence Data Analysis 2009; 13:703-723. 

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Approach. Chemometrics and Intelligent Laboratory Systems 2010; 

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2010August 11) 

100. Zadora G. Estimation of Evidence Value of Refractive Index: Influence of 

Selection of Proper Databases 2009. 

101. Zadora G, Neocleous T. Comparison of Methods of Database of Glass 

Objects Creation – Problem of Within Object Variability Estimation 2009. 

112

Examination of Paint 


Maureen J. Bradley, Ph.D. 1,2 

Andria Hobbs Mehltretter, MS 1 

Diana M. Wright, Ph.D. 1 

1 Federal Bureau of Investigation 

Laboratory Division 

2501 Investigation Parkway 

Quantico, VA 22135 

2 Author responsible for correspondence. 

113


2.  AUTOMOTIVE COATINGS  117  

2.1  Automotive Industry  119  

2.1.1  Alfa Romeo  120  

2.1.2  Audi  120  

2.1.3  BMW  120  

2.1.4  Brilliance China  121  

2.1.5  BYD  121  

2.1.6  Chrysler  121  

2.1.7  Daimler  122  

2.1.8  Fiat  122  

2.1.9  Ford  122  

2.1.10  General Motors (GM)  122  

2.1.11  Hyundai  124  

2.1.12  Honda  124  

2.1.13  Isuzu  124  

2.1.14  Kia  125  

2.1.15  Mazda  125  

2.1.16  Mercedes Benz  125  

2.1.17  Mini  125  

2.1.18  Mitsubishi  125  

2.1.19  Nissan  126  

2.1.20  Porsche  126  

2.1.21  Opel  126  

2.1.22      Saab  126  

2.1.23  Scion  126  

2.1.24  Subaru  126  

2.1.25  Suzuki  126  

2.1.26      Tata  127  

2.1.27  Think  127  

2.1.28  Toyota  127  

2.1.29  Volkswagen  128  

2.1.30  Volvo  128  

2.2  Automotive Colour Popularity  128  

2.3  Decorative Film Laminates  128  

3.  PIGMENTS  128  

4.  ADDITIVES  130  

5.  ARCHITECTURAL PAINTS  130  

6.  MARINE AND PROTECTIVE COATINGS  131  

7.  THE NATIONAL ACADEMIES’ REPORT ON FORENSIC SCIENCE (NAS REPORT)  132  

114

8.  FORENSIC PAINT STUDIES AND SIGNIFICANCE ASSESSMENTS  133  

9.  STUDIES REGARDING AUTOMOTIVE FINISHES AND REFINISHES  135  

10.  SPECTROSCOPIC METHODS  136  

11.  ELEMENTAL ANALYSIS  137  

12.  MASS SPECTROMETRY TECHNIQUES  139  

13.  ART AND CONSERVATION APPLICATIONS  139  

14.  FOURIER TRANSFORM INFRARED SPECTROSCOPY (FTIR)  140  

15.  RAMAN SPECTROSCOPY  141  

16.  FLUORESCENCE SPECTROSCOPY  143  

17.  TERAHERTZ SPECTROSCOPY  144  

18.  X‐RAY BASED TECHNIQUES  144  

19.  NUCLEAR MAGNETIC RESONANCE (NMR)  146  

20.  LASER INDUCED BREAKDOWN SPECTROSCOPY (LIBS)  146  

21.  LASER ABLATION INDUCTIVELY COUPLED PLASMA – MASS SPECTROMETRY        

  (LA‐ ICP‐MS)  147  

22.  GAS CHROMATOGRAPHY  147  

23.  CAPILLARY ELECTROPHORESIS (CE)  149  

24.  LIQUID CHROMATOGRAPHY (LC)  149  

25.  MATRIX‐ASSISTED LASER DESORPTION IONIZATION ‐ TIME OF FLIGHT MASS   

  SPECTROMETRY (MALDI‐TOF)  149  

26.  INFUSION MASS SPECTROMETRY  150  

115

27.  IMAGING (TOMOGRAPHY)  150  

28.  DETERMINATION OF MATERIALS AND ARTISTIC TECHNIQUES  150  

29.  CONSERVATION, DEGRADATION AND FORGERY DETECTION  150  


116


This is a review of the paints and coatings literature that has been published 

between September, 2007 and July, 2010 and it is divided into three sections. 

The first section addresses innovations and changes in the paint and coatings 

industry. By having knowledge of, and staying apprised of the changes in 

coatings formulations and the coatings industry, the forensic paint examiner will 

be better equipped to assess the significance of an association in comparative 

examinations. Further, with regard to the automotive industry, this knowledge 

can aid in determining the potential manufacturer, model, and model year(s) of 

evidentiary paint samples. 

2. Automotive Coatings 

Published in 2008, Automotive Paints and Coatings, edited by Steitberger and 

Dössel, is an excellent reference.[1] This is a revised and expanded version of 

Fettis’ edition of the same name that was published in 1996. Contributing 

authors provide their expertise related to all aspects of a typical automotive 

original equipment manufacturer (OEM) paint system. Historical and up-to-date 

information regarding substrate pretreatment, electrocoats, primer surfacers, 

basecoats, and clearcoats are covered in depth. In addition, topics such as inline 

repair and coatings for plastic parts to include paint, molded in color, and foil 

technology are covered. 

A number of innovations have been developed for application in automotive 

coatings. Some of the technology has already been implemented by the 

automotive industry within this review period 

As reported in the 2007 paint literature review, Volkswagen plants in Spain and 

Mexico are utilizing DuPont EcoConcept technology. EcoConcept is a patented 

finishing system that eliminates one complete coating layer, the primer surfacer, 

and combines it into a single environmentally-compliant water-based basecoat. 

As a result, both a spray booth and a curing oven are eliminated from the 

automobile production line. In 2008, German paint manufacturer Bollig & 

Kemper signed a licensing agreement for DuPont’sEcoConcept. Under this 

agreement, the Cologne-based paint maker will manufacture and sell waterbased 

basecoats for use in the EcoConcept process in automotive manufacturing 

plants in Germany and elsewhere in Europe.[2] A contact with DuPont stated 

that French automaker PSA Peugeot Citroen has been using EcoConcept since 

mid-2009 at their Poissy, France plant. Future use will be at the VW 

Chattanooga, TN plant beginning in early 2011 and several other VW plants in 

China, Brazil, and Europe possibly later that year.[3] 

Ford paint shops in India, Romania, Mexico, and China are in the process of 

converting their lines to Ford’s exclusive 3-Wet paint technology. This 

technology allows three layers of paint to be applied to the vehicle without the 

117

necessity of drying steps in between. The paint shops are more efficient, 

smaller, and use less energy. Further, the paints are high-solids; therefore, 

compared to previous formulations, less paint is needed to cover the vehicle. 

The technology was launched in 2008 at Ford’s Avon Lake, OH assembly plant 

which makes the Econoline van.[4;5] Contacts at Ford Motor Company confirm 

that the Cuautitlan, Mexico plant, which makes the Fiesta, began to use the 

process in April, 2010. Further, Ford expects to use the 3-Wet process at its 

Wayne, MI assembly plant for the new Ford Focus in December, 2010 and at the 

Louisville, KY assembly plant when a new product is launched in October, 

2011.[6] 

PPG Industries won a 2010 Automotive News PACE award for their Super High 

Power Electrocoat. This innovation offers a more uniform coating than traditional 

electrocoats, thereby reducing usage by up to one quarter. It was reported that 

PPG and Toyota have launched this “super-high-throw electrocoat” at 11 plants 

in North America and Europe.[7] Contacts within PPG stated that it was actually 

10, not 11, plants that began conversion to this process in 2008. The plants are 

in Baja, Mexico, the UK, two plants in Georgetown, KY, two plants in Cambridge, 

Ontario, Canada, two plants in Princeton, IN, and the passenger and truck 

NUMMI plants in Fremont, CA which have since closed. According to PPG, there 

is nothing that chemically distinguishes this coating from traditional ecoats. 

Rather, the innovation is attributable to the film thickness distribution of the 

product when comparing the inside versus the outside surface of the vehicle.[8] 

During this review period, both Henkel and PPG developed alternatives to 

traditional zinc-phosphate pretreatments that reduce energy consumption and 

waste. PPG earned one of R&D’s 100 Awards from R&D Magazine for their 

Zircobond ® pretreatment. The material utilizes zirconium and a proprietary blend 

of additives in place of zinc phosphate.[9] It has the same anti-corrosion and 

paint adhesion properties as zinc phosphate but reduces the sludge byproduct by 

at least 80 percent. Further, it does not generate environmentally-regulated 

heavy metals such as zinc, nickel, and manganese.[10] It can also be used in 

existing pretreatment lines. CAMI, a joint venture of Suzuki Motor Corporation 

and General Motors, began using Zircobond ® at their Ingersoll, Ontario, Canada 

plant in January, 2008. 

Henkel’s Bonderite ® TecTalis coating is also a non-phosphate conversion 

coating. It differs from the traditional zinc phosphate process in that it has fewer 

process steps and the baths do not need to be heated. After several trials at its 

Twin Cities manufacturing plant in St. Paul, MN, Ford started using the Henkel 

product on the Ford Ranger.[11-13] Kögel Fahrzeugwerke, one of the major 

manufacturers of semi-trailers in Europe, also began using TecTalis in early 

2008.[14] 

Henkel has developed a water-based automotive deposition coating that set new 

industry precedent by displacing both conventional metal pretreatment and the 

118

electrocoat process for an entire automotive vehicle body. Their Aquence 930 

coating is an epoxy-acrylic urethane coating that performs similar to cathodic ecoat 

and zinc phosphate pretreatment in automotive manufacturer cyclic 

corrosion tests. Further, Henkel was awarded a 2010 Automotive News PACE 

award for its new Aquence Co-Cure coating process. The process layers the 

Aquence autodeposition coating with a powder or liquid top coat and “co-cures” 

them together in a single bake cycle. The first dedicated commercial Co-Cure 

line was installed at Valley Towing Products in Lodi, CA which specializes in the 

manufacture of trailer hitches.[15] 

DSM Powder Coating Resins (Zolle, The Netherlands) has developed a 

corrosion-protective primer for steel that does not require a phosphate 

pretreatment. In addition to reducing production space and the necessity of 

handling and disposing of hazardous phosphate chemicals, Uralac ® Corres is 

comprised of 100% solids and therefore, solvent-free.[16] 

Santos et al. report on two alternatives to the conventional electrodeposition 

primer. One, Grancoat ZE, is an organic, zinc-rich silicate applied at a thickness 

of 4 µm. The other, Granocoat X, is a very thin (near 2µm) water-based primer 

filled with graphite. The authors found that these two products were suitable 

alternatives to current electrodeposition primers. Of interest, they also used the 

Bonderite ® pretreatment discussed previously.[17] 

Schaeffer and coauthors have developed an alternative to solvent-based 2K 

polyurethane systems. Solvent-based 2K polyurethanes are the most common 

urethane coating utilized in automotive applications. The formulations they 

studied utilize aliphatic rather than aromatic diisocyanates. The latter are 

susceptible to photo degradation. Further, the coatings are UV-cured and 

contain no hindered amine light stabilizers (HALs). The new formulations were 

tested according to strict automotive coating standards and they outperformed 

traditional 2K systems.[18] 

2.1 Automotive Industry 

The automotive industry is in constant flux. Each year, vehicle manufacturers 

phase out different makes and models and introduce new ones. They 

consolidate production to certain assembly plants, cease production at others to 

either retool the facility to accommodate the manufacture of different models, or 

close the assembly plant altogether. Since the last review period, even more 

significant changes have occurred within the automotive industry. Many were 

prompted by the economic downturn that began in 2008, which particularly 

affected the automotive industry. What follows is a summary of assembly plant 

changes, assembly plant closures, new models, new manufacturers, dissolution 

and formulation of new partnerships, and discontinued models that occurred 

during this review period. The information is listed by vehicle manufacturer. 

119

The majority of the information was obtained from Automotive News, which is a 

weekly publication. As such, some of the projections (e.g. model year available) 

may not be accurate or may not have come to fruition. Where possible, most of 

the information reported herein was confirmed via internet searches or through 

industry contacts. 

One trend that has remained steady regardless of the global economy is that car 

production in China is growing four times faster than the rest of the world. In a 

study published in 2008, it states that Chinese vehicle production more than 

doubled over the period from 2001-2005.[19] It was also reported that all major 

multinational producers of automotive coatings are active in China, supplying 

leading international automotive manufacturers, which have been setting up and 

acquiring plants in that country.[20] 

2.1.1 Alfa Romeo 

Alfa Romeo reported its new Giulietta hatchback will go on sale in Europe in 

spring, 2010. It replaces the 147, which launched in 2000.[21] 

2.1.2 Audi 

In February, 2009, Karmann stopped building the Audi A4 Cabriolet convertible, 

which had been assembled at their plant in Rheine, Germany. Karmann later 

filed for bankruptcy in April 2009.[22] In 2010, Audi began production of a fuelefficient 

premium small car, the A1. It will be available as a 2011 model in 

Europe and will be assembled at Audi’s Brussels factory.[21] 

2.1.3 BMW 

The last Z4 coupes and roadsters were built at BMW’s Spartanburg, SC plant in 

late August, 2008. The second-generation Z4, beginning model year 2009, are 

built at BMW’s Regensburg, Germany plant.[23] In December, 2008, BMW 

reported that the $750 million expansion of their Spartanburg, SC factory is 

proceeding on time and on budget. Production of the X3 crossover, currently 

built at contract assembler Magna Steyr’s factory in Graz, Austria will move to 

Spartanburg upon completion of the expansion. It is expected to begin 

production in early 2011.[23] The Dürr Group, based in Stuttgart, Germany, is 

responsible for the remodel and expansion of the paint shop at the factory. The 

production capacity is expected to increase from 160,000 to 240,000 units by 

2012. The paint shop, built by Dürr in 1996 will be expanded by 28,000 square 

meters or 80%. It will include dip-coating and drying, where RoDip (a rotation 

dipping process) will be used. Most recently Dürr expanded and modernized the 

paint shop at the BMW site in Oxford, England where the Mini is produced.[24] 

In June, 2009, the Spartanburg plant reported that about 75 percent of its 

production is exported to global markets.[25] 

In April, 2010, contacts at the BMW Spartanburg, SC plant relayed the following 

information regarding the integrated paint process (IPP II), currently being 

implemented at the plant: color codes M475, M354, and U300 (Black Sapphire 

120

metallic, Titanium Silver metallic, and Alpine white, respectively) have been 

running in limited quantities since the beginning of 2010. WA76 (Deep Sea Blue 

metallic) went into production in limited quantities in March, 2010. Colors WA68 

(Platinum Grey) and U668 (Black II) will be going into production later in 2010. 

The IPP II process, which replaces the primer surfacer with a wet on wet dual 

base coat, is running concurrently with the traditional painting operation. With 

the exception of WA68, the other five color codes are available on each of the 

vehicles manufactured at the Spartanburg plant, the X3, X5, X6, and X6 hybrid. 

Color code WA68 is available on the X5 only.[26] 

BMW’s 5-series Gran Turisimo and the X1 crossover debuted in 2010.[27] 

2.1.4 Brilliance China 

In April, 2010, Brilliance China halted exports to Europe after its BS4 and BS6 

sedans failed to gain a foothold. Combined sales of the two mid-sized sedans 

were only 502 units.[28] 

2.1.5 BYD 

The F3 Dual Mode, or F3DM for short, is Chinese automaker BYD’s first plug-in 

hybrid. In 2008 BYD reported that they sell gasoline cars to more than 20 

countries, mainly Russia and countries in the Middle East, Africa, and South 

America. At that time, they planned to continue to export conventional gasoline 

cars to developing countries and new-energy cars such as plug-in hybrids and 

electric cars to Europe and the United States in 2010 or 2011. At first, BYD’s 

cars will target mainly corporate or institutional clients such as taxis and courier 

companies in the US and Europe.[29] The F3 compact was the top-selling car in 

China in 2009. It plans to launch two cars in Europe in 2011.[30] 

2.1.6 Chrysler 

Chrysler LLC closed its St. Louis South plant (Fenton, MO) indefinitely in late 

October, 2008. The plant built the Chrysler Town & Country, Dodge Caravan, 

and export-only Chrysler Voyager. Chrysler will still build minivans in Windsor, 

Ontario, Canada.[31] Chrysler LLC’s Newark, DE plant closed in December, 

2008. The Chrysler Aspen and Dodge Durango were produced at this plant.[32] 

Chrysler plans for the Durango to return in MY2012 on a shared platform with the 

Jeep Grand Cherokee. No plans to keep the Aspen were reported. 

In mid-2009, Chrysler announced their plan for the following plant closures: St. 

Louis North, Fenton, MO (Dodge Ram – product ends December, 2009); Conner 

Avenue Assembly Plant, Detroit, MI (Dodge Viper) closing December, 2009; 

Sterling Heights, MI (Dodge Avenger, Chrysler Sebring, and Sebring convertible 

– product ends December, 2010.[33] In a conflicting report, Chrysler Group 

stated that they will continue making and selling the Dodge Viper through 

2010.[34] 

121

2.1.7 Daimler 

In August, 2009, Daimler had not confirmed reports that it may develop a new 

generation Smart ForFour with the French automaker Renault. The last ForFour 

was based on the Mitsubishi Colt and was produced at the Renault NedCar plant 

in Born, Netherlands. Due to low sales, Daimler stopped building the ForFour in 

2006 after only two years of production.[35] 

2.1.8 Fiat 

Fiat Group will stop making cars at its Termini Imerese plant in Sicily, Italy in 

2011.[36] 

Chrysler Group has announced it will retool its Toluca production plant in Mexico 

to start production of the compact Fiat 500 in December, 2010. The company 

plans to produce 130,000 vehicles in 2011, half of which will be sold as Fiat 

models in the US. The rest will be produced for the Latin American market, 

particularly Brazil.[37] 

2.1.9 Ford 

In November, 2008, Ford Motor’s Michigan Truck plant in Wayne, MI ended 

production of the Ford Expedition and Lincoln Navigator. In late March, 2009, the 

SUVs began production at the Kentucky Truck plant in Louisville, KY.[38;39] In 

spring, 2009, Ford halted production of the Mercury Sable sedan and the Ford 

Taurus X (crossover) at its Chicago, IL assembly plant.[40;41] 

Ford announced that the Ford Explorer Sport Trac pickup and Mercury 

Mountaineer SUV will be discontinued in 2010 when the Explorer, currently 

produced at the Louisville, KY plant will be moved to Ford’s Chicago plant in late 

2010. The Louisville plant will be retooled to build small cars for Ford.[42] In late 

2011, Ford plans to start assembly of the next generation of the Kuga crossover 

vehicle in Louisville, KY. Production would include US as well as European- 

model Kugas for export. The Kuga crossover is currently manufactured in 

Germany for sale in Europe.[43] 

Ford began building the Ford Fiesta in late April, 2010 in Cuautitlan, Mexico. The 

plant had been down since the end of 2008 for changeover.[44] As reported 

previously in the Automotive Coatings section, a contact at Ford Motor Company 

confirms that this plant utilizes Ford’s 3-Wet paint technology.[6] 

As was also reported in the Automotive Coatings section, Ford Motor Company 

began using Henkel’s Bonderite ® TecTalis coating as an alternative to zinc 

phosphate at their Twin Cities manufacturing plant in St. Paul, MN in 2008.[13] 

2.1.10 General Motors (GM) 

GM’s expected sale of Saturn to Penske Automotive Group was canceled on 

September 30, 2009. Within 24 hours, GM built the last Outlook crossover, Aura 

sedan, and Vue small crossover at its Lansing Delta plant in Delta Township, MI, 

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its Fairfax plant in Kansas City, KS, and its plant in Ramos Arizpe, Mexico, 

respectively.[45] 

Also in 2009, GM ended production of their Pontiac line.[46] The last Pontiac 

Torrent crossover was built at the CAMI Automotive Inc. plant in Ingersoll, 

Ontario, Canada in May, 2009. CAMI Automotive Inc. was a General Motors- 

Suzuki joint venture.[47] GM closed its Wilmington, DE plant in late July, 2009 

where it built the Pontiac Solstice.[48] The last Pontiac Vibe was built in August, 

2009 at the New United Motor Manufacturing Inc. (NUMMI) plant in Fremont, CA. 

NUMMI opened in 1984 as a joint venture between GM and Toyota Motor 

Corporation.[49] GM’s Orion Township, MI plant built its last Pontiac G6 in 

November, 2009. The plant is being retooled to begin building a new small car in 

2011.[50] GM said it will stop building the Pontiac G6 GXP sedan and coupe at 

the end of MY 2009 as well as the Chevrolet Impala SS and Cobalt SS sedans 

and the Cadillac STS-V.[51] 

GM also reported that it will no longer make the Chevrolet Trailblazer, GMC 

Envoy, or Saab 9-7X, which were assembled at the Moraine, OH plant. As a 

result, that plant closed in December, 2008. GM also ended production at Line 1 

of its Janesville, WI plant in December, 2008 which produced the Chevrolet 

Tahoe and Suburban, as well as the GMC Yukon and Yukon XL. Production of 

these SUVs will be moved to the Arlington, TX plant.[32] General Motor’s 

Oshawa, Ontario, Canada truck plant built its last truck in May, 2009 and closed. 

The plant had built the GMC Sierra and Chevrolet Silverado 1500 extended and 

crew cab pickups.[52] 

GM’s Spring Hill, TN plant closed in late 2009; it is a former Saturn assembly 

plant.[53] The plant continued to build the Chevrolet Traverse until late 

November, 2009.[54] GM moved production to its Delta Township, MI plant in 

early March, 2010. The GMC Acadia and Buick Enclave are also built at the 

Delta Township plant.[55] 

In April, 2010, GM announced the official end to the Hummer. A plan to sell the 

commercial Hummer to Chinese company Sichuan Tengzhong Heavy Industrial 

Machinery Co. fell through in February, 2010. Production of the Hummer H3 

SUV and H3T pickup at the Shreveport, LA plant ceased in January, 

2010.[56;57] 

At the end of June, 2010, the Chevrolet Cobalt ended its production run of nearly 

six years at the Lordstown, OH plant.[58] GM began building its GMC Terrain in 

August, 2009 at their CAMI automotive plant in Ingersoll, Ontario, Canada where 

the Chevrolet Equinox is also built.[59] GM will build its Chervolet Volt at its 

Detroit-Hamtramck, MI plant. Pre-production was expected to begin in March 

2010.[60] GM will build its redesigned Chevrolet Aveo (MY2011) small car at its 

plant in Orion Township, MI. The current Aveo is built in South Korea.[61] 

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The Chevrolet Cruze will replace the Cobalt in the US in 2010. It went on sale in 

South Korea in 2008 as the Daewoo Lacetti Premiere. Exports from the South 

Korean factory to Europe and Australia began in early 2009. In Australia, it is the 

Holden JG Cruze. Local production of the Cruze at Holden’s Elizabeth, South 

Australia plant is expected to begin in March, 2011. Production of the North 

American version is expected to begin in mid-June, 2010 at GM’s Lordstown, OH 

plant.[49,62:63] 

2.1.11 Hyundai 

The Equus luxury sedan debuted in South Korea in 1999. A second-generation 

model went on sale there in March, 2009 and in the People’s Republic of China 

in August, 2009. Hyundai plans to bring it to the US in August, 2010.[64] 

Production of the Hyundai Entourage minivan began in April, 2006 and ended in 

the summer of 2009 due to low sales. It was produced at Kia’s Sohari plant in 

South Korea alongside the Kia Sedona.[65] Hyundai’s 2010 sports car, the 

Genesis coupe, went on sale in the US in February, 2009. Built at the Ulsan, 

South Korea plant, it was released in October, 2008 for the Korean market.[66] 

2.1.12 Honda 

Honda manufacturing in Lincoln, AL began building the Honda Ridgeline in 

February, 2009. The pick-up had been built in Honda Motor Co.’s Alliston, 

Ontario, Canada plant.[67] In December, 2009, Honda’s Alliston Ontario plant 

began building the 2010 Acura ZDX, a mid-side luxury crossover. The plant also 

produces the Acura MDX and Honda Civic.[68] 

Honda began building its 2009 Honda Accord V-6 at its Lincoln plant in July, 

2009. It had been built at the Marysville, OH plant. The V-6 Accord is the first 

car to be built at the plant which also builds the Odyssey minivan, Pilot 

crossover, and Ridgeline pickup. The Honda Accord V-4 will continue to be built 

at the Marysville plant.[69] 

Honda’s Greensburg, IN plant began producing the Civic in October, 2008. With 

this plant online, the East Liberty, OH plant will increase production of the CR-V. 

About 40 percent of the CR-Vs sold in the US in the first nine months of 2008 

were built at the East Liberty plant; the remainder were imported from Japan and 

Mexico.[70] Honda introduced the Accord Crosstour in model year 2010 and 

deleted the S2000 sports car from its lineup.[71] Honda began building the 2010 

Accord Crosstour in November, 2009 at the East Liberty plant, which also builds 

the CR-V and Element.[72] 

2.1.13 Isuzu 

Once a lucrative light-duty vehicle in the US, Isuzu announced in 2008 that it was 

pulling out of the US passenger vehicle market in February, 2009.[73] Isuzu 

Motors America stopped delivering new vehicles to dealerships in North America 

in late January, 2009.[74] 

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2.1.14 Kia 

Korean automaker Kia is replacing its best selling Spectra with its first coupe, the 

Forte. The Forte debuted in South Korea in August, 2008.[75] It went on sale in 

the US in June, 2009.[76] The youth-oriented five-door hatchback, Soul, went on 

sale in February, 2009.[77] In 2009, Kia announced plans to cease production of 

their Sedona (Carnival) minivan in 2011.[65] Kia built a $1 billion assembly plant 

in West Point, GA, its first in North America, which will produce the 2011 model 

year Sorrento compact SUV.[78] 

2.1.15 Mazda 

The Mazda2 has been on sale in Europe and Japan since 2007; in Japan it is 

known as the Demio. It is expected to launch in the US in fall 2010.[79] 

2.1.16 Mercedes Benz 

In June, 2009, the last Mercedes-Benz CLK convertible was produced by 

Karmann in Osnabrück, Germany. Karmann filed for bankruptcy in April 

2009.[22;80] 

The first European hybrid, the Mercedes Benz S400 Blue Hybrid sedan went on 

sale in August, 2009; the ML450 SUV hybrid was launched in late 2009. The 

GLK compact SUV debuted in 2009 and the SLS AMG coupe was expected to 

debut in 2010.[27] 

In September, 2009, Daimler AG pulled the Sprinter from the Dodge lineup and 

moved the diesel-powered van to Mercedes. Dodge dealers began selling the 

Dodge-badged Sprinters in the US in 2003. Everywhere else in the world the 

Sprinter has been marketed under the Mercedes-Benz brand. The Sprinter is 

assembled at a plant in Ladson, SC.[81] 

2.1.17 Mini 

Mini has added a four-door SUV to its lineup, the Countryman. It will be 

available in model year 2011.[21] 

2.1.18 Mitsubishi 

Mitsubishi did not renew a contract with Chrysler and therefore, 2010 will be the 

last model year for the Mitsubishi Raider (rebadged Dodge Dakota).[82] Chrysler 

Group’s Warren Truck plant in Detroit, MI built the last Mitsubishi Raider in June, 

2009.[83] 

In late 2008, the CEO of Mitsubishi expressed a desire to halt Galant production 

in the US. In its place, he proposed production of small cars suitable for export 

at the company’s unprofitable factory in Normal, IL. Mitsubishi is also 

considering moving production of the Lancer small sedan and Outlander 

crossover to this plant; these models are sold globally.[84] 

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Under an agreement announced in early March, 2010, Mitsubishi will produce 

approximately 100,000 electric vehicles for French carmaker PSA/Peugeot 

Citroen. Mitsubishi will build the four-seat electric cars at its plant in Mizushima, 

Japan. The PSA models are based on Mitsubishi’s ultra compact i-MiEV which is 

on sale to fleets in Japan and is expected to be on sale to private buyers by 

June, 2010. The cars for PSA will get different body styling and interiors and will 

have different handling and suspension. PSA plans to have approximately half of 

the vehicles sold under each of its brands, Peugeot and Citroen.[85] 

2.1.19 Nissan 

In 2009, Nissan moved production of their Quest minivan out of Canton, MS back 

to Japan. This allows for increased production of the Altima at the Canton 

plant.[69] Nissan will begin selling its Juke, a compact crossover, in Japan in 

summer, 2010 and in the US and Europe in the fall, 2010.[86] The Juke is made 

at Nissan’s Oppama, Japan and Sunderland, UK plants. 

2.1.20 Porsche 

Porsche’s Panamera 4-door sedan debuted in calendar year 2009.[27] 

2.1.21 Opel 

Opel plans to build low-priced cars at GM’s plant in the northern Russian city of 

St. Petersburg. GM opened the $300 million plant in November, 2008 and 

currently builds the Opel Antara and Chevrolet Captiva SUVs.[87] 

In late November, 2009, GM stated that it plans to keep all four of its Opel 

factories in Germany open under a $4.92 billion restructuring plan.[88] Opel’s 

Antwerp, Belgium plant will be closed and production of the Astra HB3 will move 

to Bochum, Germany. The Bochum plant will also have exclusive production of 

the new Zafira.[89] 

2.1.22Saab 

After the sale of Saab to Swedish company Koenigsegg Group AB fell through, a 

small Dutch producer of luxury sports cars, Spyker, purchased Saab from GM in 

January, 2010.[90;91] 

2.1.23 Scion 

The iQ went on sale in Japan and Europe as a Toyota during the fourth quarter 

of 2008.[92] 

2.1.24 Subaru 

Subaru added production of their Outback wagon to their Lafayette, IN plant. 

The Legacy and Tribeca SUV are also produced at this plant.[93] 

2.1.25 Suzuki 

Suzuki ended production at the CAMI Automotive Inc. Ingersoll, Ontario plant in 

June 2009. CAMI, a joint venture with GM, made only four Suzuki XL7s in 2009. 

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The XL7 was the largest vehicle made by Suzuki and the only model 

manufactured in North America.[94] In December, 2009, Suzuki sold its 50 

percent stake in the plant to GM.[95] 

Suzuki builds its mid-sized Kizashi sedan in Sagara, Japan alongside its SX4 

subcompact car. It went on sale in Japan in October, 2009, in North America in 

December, 2009, and in Australia in May, 2010.[96] 

2.1.26Tata 

In 2009, Tata Motors (India) rolled out its tiny Nano. The Nano retails for about 

100,000 rupee ($2000 USD).[97] 

2.1.27 Think 

In January, 2010, electric-vehicle maker THINK North America announced it will 

build a manufacturing facility in Elkhart, IN and build approximately 20,000 cars 

per year through 2013. The THINK City will have recyclable plastic parts and a 

recyclable interior. It is expected to go on sale in the US in the first quarter of 

2011. The first cars to be sold in the US will be built in Finland until production 

begins in the US in 2011.[98] The THINK City was the first highway-capable 

electric vehicle certified to European safety standards. It has been on sale in 

Europe since December, 2008.[99] 

2.1.28 Toyota 

In 2008, Toyota launched the RAV4 built at its new plant in Woodstock, Ontario, 

Canada.[100] The Toyota Sai, Toyota’s first hybrid sedan, went on sale in Japan 

in October, 2009.[101] 

In 2009, Toyota ended production of the Corolla and Tacoma pickups at the 

NUMMI plant in Fremont, CA. NUMMI was a joint venture between Toyota and 

GM. The plant opened in 1984. Toyota will shift production of the Tacoma to its 

truck factory in San Antonio, TX. NUMMI’s output of the Corolla will be replaced 

by cars from Japan and Cambridge, Ontario, Canada.[102] Toyota also 

consolidated all production of its Tundra truck to San Antonio.[69] 

Toyota’s plan to build a $1.3 billion Prius factory in Blue Springs, MS stalled in 

2008 due to the economic recession.[103] In June, 2010, Toyota announced 

plans to resume construction on the plant and begin producing the Corolla there 

by the end of 2011. This plan would move production of the Corolla back to 

North America from Japan. 

The 2010 Toyota Highlander crossover began production at the Toyota Motor 

Manufacturing plant in Princeton, IN in October, 2009. Toyota also makes its 

Sequoia SUV and Sienna minivan there.[104] It continues to import Highlanders 

produced at their Kyushu, Japan plant.[105] 

127

Toyota owns 8.7 percent stock in Fuji Heavy Industries, Subaru’s parent 

company. As such, in February, 2007, Subaru of Indiana (Lafayette, IN) began 

producing the Toyota Camry.[93;106] 

2.1.29 Volkswagen 

Chrysler assembles the Routan for VW at its Windsor, Ontario, Canada minivan 

plant alongside the Chrysler Town and Country and Dodge Grand Caravan.[107] 

In early 2010, Volkswagen will begin global marketing for the Amarok, a compact 

pickup truck built in Argentina.[108] 

2.1.30 Volvo 

Swedish brand Volvo debuted its XC60 crossover in calendar year 2010.[27] It 

has also introduced the S60, a four-door coupe. The company says it plans 

yearly global sales of the new S60 to be about 90,000.[21] 

2.2 Automotive Colour Popularity 

According to annual automotive colour popularity data compiled by paint 

suppliers DuPont and PPG, silver, black, and white continue to be the most 

popular colors; blue and red round out the top five. Depending on the country or 

continent (e.g., North America, Europe, or Asia), the rankings of the top five 

colors fluctuate somewhat.[109;110] 

2.3 Decorative Film Laminates 

The automotive industry is the largest user of decorative film laminates. Poole of 

Soliant (Lancaster, SC) provided an excellent overview of how decorative film 

laminates are utilized in pressure sensitive decals, extrusion lamination, in-mold 

decorating, thermoforming, and insert injection molding.[111] Suppliers that 

specialize in certain parts do not have to worry about a paint match because 

different suppliers can use the same paint film. Uses such as a chromeappearing 

decorative film can provide the look of a metal chrome bumper without 

the environmental concerns of chrome-plating. Further, the reduced weight of 

the vehicle when a film substitutes for metal translates to higher fuel efficiency. 

Soliant’s Fluorex ® Paintfilm was selected for the 2008 Jeep Wrangler Sahara 

front and rear bumper program. The paint film is supplied as a thick sheet 

laminate for thermoforming. Fluorex ® provides a high-gloss Class A finish with 

outstanding stone and chip resistance, weather resistance, and enhances impact 

resistance.[112] 

3. Pigments 

Pigment manufacturers have brought a number of innovative products to market 

in the last three years. Silberline (Tamaqua, PA) introduced several new metallic 

effect pigments for automotive coatings. The Platinum Silver product series 

provides a wide range of metallic effects from bright silver to chromatic. The 

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newest edition to the Sparkle Silver Ultra ® line is SSU 6656, which is highly 

polished, extremely bright and lenticular in shape (convex on both sides).[113] 

Sun Chemical (Parsippany, NJ) launched a new portfolio of metallic pigments 

called SunMetallics ® . It is a line of non-leafing aluminum flake pigments for use 

in solvent-borne coating applications. The product line includes corn flakeshaped 

and silver dollar-shaped pigments over a broad particle size range. 

SunChemical also introduced another effect pigment product line, SunGem ® . In 

addition to high chroma color, excellent opacity, and a combination of 

pearlescent and metallic character, it has unique magnetic properties that allow 

for the ability to control pigment placement.[114] Further, SunChemical recently 

launched Polymer Encapsulated Aluminum Pigments. They were first introduced 

in China for use in consumer electronics, automotive trim, and other single coat 

applications that require a high level of acidic and base resistance. They are 

designed to maintain the brilliance and gloss of the base pigments while the 

polymer encapsulation protects them from exposure to acid or basic materials. 

Further, by insulating the pigments, their conductivity is reduced, a plus for use in 

consumer electronics.[115] 

In 2009, BASF launched Glacier Frost White effect pigment which is the 

newest generation of synthetic mica coated with titanium dioxide. Since the 

pigment’s substrate does not contain impurities, the result is a very clean color. 

According to BASF, “…it has been referred to as the ‘whitest white’ pearlescent 

pigment in today’s market”.[114] BASF has also introduced Paliocrom ® Brilliant 

Orange, an iron-coated aluminum pigment. It has high chromaticity, excellent 

hiding power, and inherent sparkle. According to BASF, “…it has been 

especially well received by the automotive community and is currently offered in 

new red, orange, and gold color stylings”.[115] 

Ciba (now part of BASF) introduced XYMARA Fireball, which combines the 

lightfastness of a typical quinacridone pigment with the luster of a mid-range 

metallic or pearlescent pigment. Prior to this, formulators would have to utilize a 

transparent high performance red with a weather resistant effect pigment.[116] 

Ciba has also introduced a line of pearlescent pigments that are coated glass 

flakes. This line is marketed under XYMARA Nordic.[117] 

Heubach GmbH (Langelsheim, Germany) has expanded its line of high 

performance pigments to include Monolite ® Yellow 113901 (PY139), Monolite ® 

Red 325401/325402 (PR 254) and Monolite ® Red 312201 (PR 112).[116] 

Shiomi and coauthors developed a pearlescent pigment in which synthetic mica 

is coated with iron oxide and then titanium dioxide. The pigment had 

approximately twice the chroma value as compared to current commercially 

available pigments. Although developed for the cosmetics industry, this pigment 

does have application in automotive coatings and molded plastics.[118] 

129

BASF introduced a new automotive coating, Liquid Metal, at the Geneva Motor 

Show held March, 2010. The water-borne basecoat used physical vapor 

deposited aluminum to achieve its unique effect. Rather than painted, the 

automobile appears to be made of chrome. It was demonstrated on the F 800 

Mercedes-Benz as Alubeam paint, and on a Hyundai concept car, the iflow.[119;120] 

BASF also introduced their Sicotan ® , Sicopal ® , and Lumogen ® 

pigments at the Geneva Motor Show on the Hyundai concept car, the i-flow. 

These pigments are a range of infrared-transparent and infrared-reflecting 

pigments. They were utilized in a number of applications in the car interior and 

resulted in cooler plastic surfaces of the dashboard, steering wheel, and center 

console.[119] 

Frischmann describes the lightfastness, excellent weatherability, and exceptional 

temperature resistance of Heubach GmbH’s line of HEUCODUR ® IR pigments. 

These infrared-reflective complex inorganic pigments offer a wide range of color 

shades from yellow to black and can be utilized in architectural coatings, as well 

as both interior and exterior automotive applications. Utilizing these pigments in 

automotive paint or molded plastic reduces heat build up and extends the life of 

the polymeric matrix.[121] Banerjee provides a brief overview of the principles of 

heat reflection by traditional infrared-reflective pigments, such as titanium 

dioxide, carbon black, and C.I. Pigment Black 30.[122] 

4. Additives 

Each year Paints and Coatings Industry (pci) magazine compiles an Additives 

(Guide) Handbook. It is a comprehensive list that includes definitions and 

examples in a format that is easily understood regardless of the readers’ paint 

experience. The 2010 Additives Handbook is available on-line at no charge from 

the www.pci.com website.[123] 

A comprehensive review of hydroxyphenyl-s-triazine (HPT)-based ultraviolet 

absorbers (UVAs) and their current applications in coatings for plastics and 

wood, as well as powder coatings, is provided by Schaller and coauthors.[124] 

In their research, Lowry and coauthors demonstrated that nearly complete UV 

protection can be achieved in clear coats utilizing ZnO nano-particles.[125] 

5. Architectural Paints 

Hollow latex particles (HLPs) are a new class of raw material which combine the 

binding function of the conventional latex binder with the pigment functions of 

traditional light-scattering pigments such as titanium dioxide.[126] HLPs can be 

the sole binder in a paint and allow for a significant reduction in titanium dioxide 

usage. Paints made with HLPs offer properties comparable to those made from 

conventional binders and pigments with the added benefit of lowering raw 

material costs. At this point of development, HLPs have low gloss potential, so 

they are limited to flat paints. 

130

6. Marine and Protective Coatings 

In their article “Anticorrosive coatings: a review” Sorensen and coauthors provide 

an excellent description of the utility and composition of anticorrosive coating 

systems.[127] This reference is particularly helpful to the forensic paint examiner 

because these types of coatings are not routinely encountered in case work. The 

authors’ review includes approximately 350 references as well as comprehensive 

descriptions and examples of the binders and pigments utilized in these coatings. 

In addition, protective mechanisms of barrier, sacrificial, and inhibitive coatings 

are described in detail. 

In their research, Naderi and Attar evaluated the effectiveness of zinc aluminum 

phosphate in comparison with zinc phosphate as an inhibitive pigment.[128] 

Modification of the cationic constituent resulted in superior performance as 

compared to zinc phosphate. 

Silicone-organic resin systems, particularly silicone-epoxy resins, have 

established a market segment in the protective coatings industry. Compared to 

the industry standard of three- to four-coat 2K urethane systems, silicone-epoxy 

hybrid technology offers reduction in VOCs, elimination of isocyanate health 

effects, and improved durability against weathering, corrosion, and chemical 

attack.[129;130] 

Antifouling coatings are used to prevent organisms such as barnacles from 

growing on a ship’s surfaces, which can increase fuel consumption by as much 

as 40%. In addition to fuel savings, a reduction in millions of tons of CO2 and 

SO2 emissions can also be realized.[131] Currently, cuprous oxide (Cu2O) is the 

antifoulant most widely used. Organic biocides are also applied to supplement 

the antifouling effect. As copper prices continue to rise, marine coatings 

companies have launched a number of new products that are more 

environmentally friendly and increase fuel efficiency. International Paint 

introduced Intersleek ® 900, a biocide-free fluoropolymer foul release coating in 

2007. They have since introduced a number of high volume solids (78-80%) 

primers.[132] Hempel (Kongens Lyngby, Denmark) has introduced Hempasil X3 

which uses hydrogel silicone to prevent fouling on a vessel’s hull.[133] The 

water absorbent hydrogel forms a polymeric network over the surface such that 

organisms perceive the hull as a liquid, not as a solid surface. As a result, the 

organisms do not try to latch on.[134] Nippon Paint Marine Coatings, Co. 

(Nagata-Ku, Japan) has developed a marine paint that contains a proprietary 

polymer compound that turns to a gel when it comes into contact with water. The 

resulting smooth surface causes a significant reduction in drag.[135] 

Sherwin-Williams Industrial and Marine coatings has introduced SeaGuard 6200, 

a high solids (80%) epoxy that is free of heavy metals.[131;134] Researchers in 

Denmark are also evaluating the potential use of peroxides of magnesium, 

131

calcium, strontium, and zinc as active ingredients in antifouling coatings. 

Hydrogen peroxide and highly water-soluble ions would form when the coating is 

exposed to water, with the hydrogen peroxide then serving as the biocide.[136] 

The next section in this paint literature review discusses forensic paint analysis in 

the traditional sense, in that the topics include frequency of occurrence studies, 

discrimination by various analytical techniques, and the conclusions that can be 

drawn from each. 

7. The National Academies’ Report on Forensic Science (NAS Report) 

Arguably the most significant development that occurred during this review 

period was issuance of the report “Strengthening Forensic Science in the United 

States: A Path Forward” by the National Academies based in Washington, 

DC.[137] This organization assembled a panel of subject matter experts to 

review the status of forensic science in America through the use of interviews 

and research by the panel’s sub-groups. Paint analysis was one of the areas 

studied and discussed in the final report. 

The major recommendation for paint examinations was repeated for all 

disciplines. Overall, the panel stressed the need for better standardization of 

definitions across laboratories in report writing, interpretation of results, and 

significance assessments. The need to include sources of uncertainty was also 

stated. The basic recommendation seems to suggest that forensic scientists 

should seek the development of laboratory reports in which the findings would be 

clear and thorough without the need for testimony to interpret the results. To that 

end, the Bundeskriminalamt, the FBI Laboratory, and several state laboratories in 

North America have begun to include an interpretation of results section in their 

reports of examinations. For example, the BKA and the FBI Laboratory’s Paint 

group have each developed associative scales that are included for interpretation 

of their reports. These models were based in part on presentations and 

discussions with Christopher Bommarito of the Michigan State Police Laboratory 

system in Lansing, Michigan. 

Further, the Scientific Working Group for Materials Analysis (SWGMAT) has 

formed a working group to determine if a standard format for interpretation 

assessments can be developed to encompass the various sub-disciplines that 

are part of trace evidence analysis (e.g., fibers, glass, hair, paint, and tape). If a 

single format is approved, SWGMAT members would determine if an 

interpretation of results guideline for trace evidence analysis could be issued, or 

whether an interpretation section would be added to the guidelines published by 

each sub-discipline. 

Other recommendations relevant to forensic paint analysis from the NAS report 

included the need for laboratory accreditation, annual discipline-specific 

proficiency testing, and certification of individual trace evidence examiners by an 

132

independent accrediting organization. Several accrediting bodies for forensic 

laboratories exist in North America and Europe. Most of these entities have 

adapted their accreditation criteria to embody the ISO/IEC 17025 standard for 

certification, the main standard used by testing and calibration laboratories. The 

European Paint and Glass group conducts annual collaborative exercises in paint 

analysis, and several independent proficiency test providers exist in North 

America. 

Also, since the last review, the ability to attain individual certification in the United 

States has become more readily available to paint examiners. As an example, 

the American Board of Criminalistics (ABC) offers certification in several 

specialized disciplines, such as trace evidence. Specifically, the ABC’s offerings 

now include a specific paints and polymers certificate, available to anyone who 

meets the acceptance criteria to achieve and maintain this certification. 

The biggest challenge to paint examinations remains the foundation upon which 

conclusions are based. Uncertainty and significance assessments are often cited 

as necessary and yet are not able to be designed as all encompassing for 

interpretative use. Yet, some have been conducted during this review period. It 

seems logical, therefore, to begin discussion of analytical applications with these 

studies as they pertain to the forensic analysis of paint, the significance of the 

results, and how researchers are attempting to apply statistics to these analyses. 

8. Forensic Paint Studies and Significance Assessments 

Prior to issuance of the NAS report, at least four studies regarding the 

significance of forensic paint examinations were underway by researchers in 

Europe and the United States. Two of these studies attempted to describe the 

significance of automotive paint analysis, and two involved analysis of 

architectural paints. All of this work was presented at the Trace Evidence 

Symposium entitled: Interpretation of Trace Evidence: The Present and Future, 

which was held in Clearwater, Florida in 2009, and is available on-line at 

http://projects.nfstc.org/trace/2009/index.htm. 

The first of the automotive paint studies was reported on by Louissa Marsh, 

where random samples of road debris were analyzed for the purpose of 

determining how common automotive paint chips would be on a road 

surface.[138] This study demonstrated that within the UK, there was a greater 

than 74% chance that paint chips exist on roadways. However, the frequency of 

occurrence was found to be only 7% for automotive paint chips. Similarly, in a 

separate study, no automotive paint was observed amongst the 1,253 paint chips 

recovered from 100 items of random clothing that were processed for debris. 

Parent and Howard of the Texas Department of Public Safety Crime Laboratory 

co-authored the second automotive paint study, in which 500 automotive paint 

samples were collected from damaged areas of vehicle hoods (bonnets) during 

133

the summer of 2007, and analyzed in order to determine if any random matches 

were possible.[139] Model years ranged from 1979 to 2007 with some years in 

the early 1980s not represented and the median model year calculated to be 

MY2000. The majority of the vehicles were manufactured in North America by 

Chrysler, Ford, or General Motors, the latter of which accounted for the greatest 

number of vehicles sampled (~160). In total, 20 of the 500 samples were not 

able to be discriminated. However, 393 were differentiated based on physical 

characteristics alone. Fourier transform infrared spectroscopy (FTIR) analysis 

discriminated 80 additional samples and pyrolysis-gas chromatography/mass 

spectrometry (Py-GC/MS) further distinguished 7 samples. The 20 remaining 

samples equated to 14 indistinguishable sample pairs, which yielded a 

discrimination value of 99.99%. The authors also reported that within these 14 

sample pairs, two pairs contained vehicles assembled at plants in different 

states. Of the remaining sixteen samples, both samples within each of the twelve 

pairs were collected off of vehicles assembled at the same plant. 

Regarding the architectural paint comparison studies, the discriminating power of 

visual and microscopic techniques in conjunction with FTIR was also 

demonstrated in the study presented by Wright et al. [140] Over 950 architectural 

paints were randomly collected across the United States and Canada. 

Approximately half of these samples contained a surface color observed to be 

white or off-white. In order to best determine the discriminating power of the 

analytical scheme, FTIR was utilized on the surface layer of the approximately 

200 white samples prior to side-by-side macroscopic and microscopic 

comparisons of the top layers. This approach enabled the authors to report a 

discriminating power of 99.99% for the combination of these techniques. 

Scanning electron microscopy with energy dispersive X-ray spectroscopy 

(SEM/EDS) and Py-GC/MS were also utilized in this study with additional sample 

discrimination, such that only 11 pairs remained undifferentiated at the 

conclusion of the study. However, the greatest significance of this work was that 

the samples contained within each of the 11 pairs were later found to be nonrandom. 

Each paint sample within a pair shared a common origin in that it was 

collected from the same source (e.g., building or structure) as the other 

sample(s) within that same pair. 

The last of the four paint significance studies presented at the Trace Evidence 

Symposium was authored by Massonnet.[141] In this work, a market study of 

commercially available red architectural paint was conducted in order to 

determine the significance of results in which two similarly colored, yet chemically 

different red paints were used to vandalize two public buildings on different days 

using the same general method of application of paint to the building surfaces. 

Traces of a visually similar red paint were also observed on a subject’s shoes, 

which were supplied to the author for comparison. The analytical data was 

collected using FTIR and Raman spectroscopy. Through the use of a Bayesian 

approach to data interpretation, the author compared the collected data to 28 red 

paint samples with the same color code and determined that the data strongly 

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supported a conclusion that the red paint on the shoes and the red paint applied 

to one of the buildings for the purposes of defacing its surface contained the 

same observed and measured characteristics. 

Two articles were published by Zadora and coauthors that described ongoing 

efforts to determine the significance of paint transfer evidence using likelihood 

ratios. One study utilized data generated by FTIR analysis and the other 

employed Py-GC/MS results. [142;143] Stable isotope analysis of white paints 

was another application of likelihood ratios described by Farmer et al. In this 

study, isotope ratio mass spectrometry (IRMS) was used to analyze 13 C, 18 O, and 

2 H isotopic compositions originating from paint binders.[144] 

Further discussion of significance assessments were reported in Science and 

Justice in 2010 as abstracts of presentations given at the 5 th Triennial 

Conference of the European Academy of Forensic Science held in Glasgow, 

Scotland in September 2009.[145] A number of presentations addressed the 

need to provide context to report conclusions. Use of Bayesian statistics, 

likelihood ratios, and development of a unified ordinal scale to describe levels of 

conclusions were presented. 

As a final note to the discussion of significance, paint examiners routinely review 

trade journals and consult with experts from the coatings industry in order to 

guide critical thinking as to the probative value of an examination request or the 

appropriate interpretation of analytical findings. In this regard, educating the 

paint and coatings industry about forensic paint examinations can be as critical 

as the reverse. To that end, Mike Eyring, formerly of the Arizona Department of 

Public Safety, wrote a review of several forensic paint cases that highlight how 

forensic paint examinations are conducted and how they can be used within the 

context of criminal investigations.[146] 

9. Studies regarding automotive finishes and refinishes 

Research on the performance of coated galvanized steel was reported on by 

Santos et al. in which waterborne and solvent borne basecoats were compared 

on primed and pre-primed steel sheets in order to study effects on 

adhesion.[147] In general, the waterborne finishes showed better adhesion 

results over the solvent borne coatings. Not surprisingly, the use of primers also 

promoted better adhesion. Other results included comparison of the differences 

noted between electrogalvanized, hot dip galvanized, and galvannealed steel for 

these adhesion tests. 

Three papers were published during this review period that discussed analysis or 

development of systems used in automotive refinishes. In the first, automotive 

body fillers were characterized in order to better assess discrimination of these 

materials in casework.[148] Thirty-three samples of body filler and spot putties 

were analyzed using light microscopy, FTIR, SEM/EDS, and Py-GC. Overall, 19 

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of the 33 samples were able to be differentiated with the authors concluding that 

SEM/EDS offered the best discrimination capability. 

The two remaining papers discussed clearcoat refinish systems and were 

published by members of the Akzo Nobel Car Refinish group in the Netherlands. 

The first paper discussed the interaction of the basecoat and clearcoats within a 

refinish system and made several observations based on testing done using 

FTIR and SEM/EDS.[149] The authors describe evidence of strong interaction 

between the clearcoat with the basecoat. They suggest that the data revealed 

isocyanurate crosslinker in polyurethane clearcoats penetrated to a large extent 

into the basecoat, even to the point of being detected at the basecoat/substrate 

interface. The authors discussed the improved mechanical properties of the 

basecoat observed in this system as attributable to an in-situ crosslinking of the 

basecoat binder, where the Tg and crosslink density of the basecoat increased. 

When the interaction of the basecoat to a UV-clearcoat was studied, the 

interaction appeared to be much less between the two layers due to the high 

cure rate. The authors state that the curing process used prevented diffusion of 

the crosslinker into the basecoat. Lastly, they noted that stabilizers such as 

hindered amine light stabilizers (HALs) and UVAs also diffused into the basecoat 

thereby providing better weathering protection. 

The second paper, authored by some of the members of this same research 

group, reported on the development of a waterborne UVA curable clearcoat 

refinish, concluding in part that the coating under development demonstrated 

good gloss retention even after two years of exposure in environmental 

conditions common to Florida.[150] 

10. Spectroscopic Methods 

Scientists at the Leibniz Institute of Surface Modification published work on the 

use of near infrared spectroscopy to monitor polymer coat curing via UV light and 

the resulting coating thickness in-line during production. The precision of the 

measurement was determined to be less than 1 micrometer at best. [151] 

Comparative studies of small samples of automotive coatings using visible 

microspectrometry were also reported on by researchers in Poland. [152;153] 

In another study, paint cross sections were characterized by Joseph, et al. using 

mapping and linear imaging capabilities with attenuated total reflectance (ATR) 

microscopy.[154] In this work, two different FTIR microscopy systems were 

studied: one with a conventional DTGS detector; and, one with FTIR mapping 

capabilities using a novel integrated system with raster scanning in ATR mode. 

The latter system was found to provide a better signal-to-noise ratio with shorter 

acquisition time when compared to the conventional system. The authors state 

that this newly developed system has features of particular benefit to the 

identification and spatial locality of thin organic layers within a paint cross section 

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as well as to low concentration organic components of mixtures that contain both 

inorganic and organic moieties. 

A further forensic application was reported by Szafarska and coauthors in which 

ATR-FTIR was used to analyze paint the authors applied to polyester and fabric 

matrices. [155] Applying a series of normalization and subtraction steps to the 

collected IR data, the authors determined that this approach would be an 

acceptable alternative to physically removing paint from a substrate for analysis. 

In a study employing the use of synchrotron radiation, eighteen titanium dioxide 

(TiO2) pigments in rutile form were examined along with fragments of seven 

types of white automotive paints. Results indicated detection of elements such 

as niobium, which is indicative of the sulfuric acid preparation method of the TiO2 

pigment. Aluminum and silicon could not be effectively excited and therefore 

detected with the described high-energy synchrotron radiation X-ray fluorescence 

(XRF) analytical scheme. They were, however, observed by SEM/EDS. For the 

white automotive paints, detection of trace amounts of heavy elements was used 

to discriminate samples that were indistinguishable by color measurement of the 

white basecoat.[156] 

Uses of Raman spectroscopy with respect to modern forensic applications was 

reported by Otieno-Alego.[157] Also, the combined techniques of µ-Raman and 

µ-XRF were applied to multilayer paint chips using a single spectrometer.[158] 

The use of confocal Raman microscopy to monitor the distribution of melamine in 

a polyester-melamine coil coating as well as for depth profiling of these coatings 

was reported on by Zhang and coauthors.[159;160] This same group of 

researchers also reported on the use of step-scan photoacoustic FTIR for the 

latter application.[161] 

Lastly, for consideration in potential sourcing of raw materials, researchers in 

Pakistan described synthesis of ultramarine blue pigment from kaolin mined in 

the Swat valley region of that country. Analysis of the end product was 

accomplished using UV-vis, FTIR, and X-ray diffraction (XRD) analysis.[162] 

11. Elemental Analysis 

Applications in the area of elemental analysis of paint samples included 

spectrometric techniques such as particle-induced X-ray emission (PIXE), XRF, 

SEM/EDS, XRD, and laser induced breakdown spectroscopy (LIBS). In one 

article, PIXE, XRD, and backscatter capabilities associated most commonly with 

SEM/EDS applications were used in tandem to simultaneously acquire data on 

pigments and binder matrices in paint layers.[163] With this research, the 

authors were attempting to resolve the problem of estimating the pigment/binder 

ratio present in paint, which is often complicated in SEM/EDS by the presence of 

carbon and oxygen in both entities. Their resolution involved a three-step 

process: determine the pigment identities and concentrations via micro-XRD and 

137

PIXE, respectively, determine the total elemental composition of the paint matrix 

to include the pigment and binder via backscatter imaging, and then subtract the 

carbon and oxygen content of the pigments from the whole in order to calculate 

the contribution from the binder alone. The authors cite that the advantage of 

this approach is the ability to analyze the same area of detail within one 

experimental set-up scheme. 

EDXRF was also used in conjunction with digital imaging and chemometrics to 

determine the weatherproofing classification level and durability of 17 varnishes 

and 10 paints subjected to accelerated weathering tests as part of a quality 

assurance test plan.[164] 

A portable XRD/XRF instrument design was discussed by Pifferi et al. for 

nondestructive analysis. Its features include the ability to acquire data using both 

techniques simultaneously for qualitative and quantitative work in a nondestructive 

mode.[165] Nakai and coauthors have published results of 

experiments conducted on the capability of SEM/EDS when coupled with a 

transition edge sensor (TES) microcalorimeter to analyze white automotive 

paint.[166] Samples were prepared as would be typical for SEM analysis in that 

chips were embedded in a resin, polished to reveal the layer structure at the 

surface of the mold, and coated with carbon. Acceleration voltages were the 

same for both the TES and traditional solid state detector (SSD) at 15 KeV. 

Beam currents were also comparable. Data acquisition time for the TES was 

300 live seconds as compared to 100 seconds for the SSD. The authors report 

that the TES detector as used in this work was able to resolve titanium 

interferences on the barium signal in the white primer layer, which also contained 

a detectable signal for sulfur. It was reported that this detection system also 

provided a better estimate of the ratio of lead to tin in the electrocoat layer. 

Additional research regarding elemental analysis of paint layers utilized direct 

sampling of acrylics for analysis by graphite furnace atomic absorption 

spectrometry (GFAAS), as well as the detection of bromine in industrial paints by 

inductively coupled plasma–mass spectrometry (ICP-MS), where it can exist as a 

flame retardant.[167;168] 

Lastly, the use of LIBS to monitor application of paints and coatings to metal 

substrates has been reported on by Kim et al.[169] In this report, a compact 

LIBS system was described for use in identifying eleven different yellow OEM 

paints that were developed to be undifferentiated by the human eye. In this 

manner, different applications would all appear to be of a uniform color. In this 

example, the color was the yellow recognized as belonging to the Caterpillar, Inc. 

heavy-machinery company. The ability to differentiate between urethane, epoxy, 

and alkyd formulations provided resolution in two cases of paint failure, which 

were able to be readily diagnosed at field sites using this technology. 

138

12. Mass Spectrometry Techniques 

Pyrolysis gas chromatography–mass spectrometry (Py-GC/MS) was the most 

common mass spectrometric technique reported on for paint analysis since the 

last review. As a means of better understanding its capabilities and results, 

Wampler and coauthors published two review articles in PCI (Paint and Coatings 

Industry) magazine. The first describes a simple way to identify polymers based 

on the collected pyrogram data for a polymeric sample.[170] The on-line version 

of this article also contains a table that summarizes pyrolyzate peaks for 100 

polymeric materials. The second article specifically discusses coatings analysis 

via Py-GC/MS.[171] Examples of the anticipated pyrolyzates obtained for 

common materials such as acrylics, epoxies, and architectural paint formulations 

are given. 

Researchers in Poland published four papers described the utility of Py-GC/MS 

for automotive paint layer analysis [172-175] and one discussed the study of 

spray paint on plasters.[176] 

A study involving the use of laser desorption mass spectrometry (LDMS) was 

also reported during this review period. Mukai and collaborators studied blue 

copper phthalocyanine pigments used in automotive coatings, furthering previous 

work in which LDMS has been demonstrated to be effective in the identification 

of these types of organic pigments.[177] As was reported in the 2007 paint 

review article, this technique is advantageous in forensic and conservation 

applications because it can be applied nondestructively to small samples in order 

to provide fast results with relatively low detection. Another advantage is the 

capability of LDMS to simultaneously detect inorganic and organic pigments at 

low concentrations within a binder matrix. 

13. Art and Conservation Applications 

Although forensic paint examiners rarely examine artworks, methodology and 

research in the practice of art conservation is quite relevant, as some of the 

analytical requirements and goals are the same: small sample sizes with minimal 

damage to the material being analyzed. Similarly, the requisite interpretation 

skills are common to both disciplines. In this review period, the publications 

generated in art conservation and analysis surpassed that of forensic sciencespecific 

research articles. However, the information gained from these studies is 

applicable across both fields. Therefore, relevant artwork publications are 

reviewed here, inasmuch as many could be of interest to forensic scientists. 

As with forensic work, methods of analysis pertaining to art conservation and 

analysis range from simple microscopical and microchemical examinations to 

highly specialized instrumentation. These latter techniques have greater limits of 

detection, significantly reduced or almost negligible sample size requirements, 

and improved sensitivity and resolution. Further, it has been observed that many 

139

of the more current studies involved analysis of artwork by multiple techniques in 

conjunction, providing a greater amount of complementary information on a 

single item than was previously possible. 

This portion of the review will therefore describe key publications covering the 

range of art and archaeological materials analysis from primarily single-technique 

examinations to analysis by multiple techniques in combination. Singletechnique 

analysis will be broken down into spectroscopic techniques (FTIR, 

Raman, fluorescence), X-Ray-based techniques (SEM/EDS, XRF, XRD, PIXE), 

laser-based atomic techniques (LA-ICP/MS, LIBS), and separation techniques 

(GC/MS, CE). 

14. Fourier transform infrared spectroscopy (FTIR) 

With regard to FTIR analyses, two primary trends were noted in this review 

period: analysis in reflectance mode and the use of multivariate statistics to aid in 

data analysis. In fact, some research incorporated both trends. In Rosi et al., 

the strengths and limitations of reflectance mid-FTIR chemometric models for 

binder identification were discussed, including how principal components 

analysis (PCA) can be used as a data analysis tool.[178] Navas and coauthors 

combined diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) 

with PCA to compare transmission FTIR to DRIFTS and found that the intensity 

was improved in DRIFTS, making it more suitable for purposes of identification 

and characterization of pigments.[179] 

Several additional publications considering multivariate statistics were of note. 

One included FTIR-ATR analysis of reference standards of polymeric resins to 

evaluate multivariate statistics (hierarchical clustering, PCA, Linear Discriminant 

Analysis (LDA).[180] LDA provided the best resolution for separating similar 

resins. Gruchow and coauthors primarily investigated transmission FTIR 

comparing univariate with multivariate statistics, demonstrating the usefulness of 

cluster analysis and PCA.[181] Schäning et al. tried to facilitate classification of 

organic pigments by partial least-squares discriminate analysis and concluded 

that the “…application of the classification models to FTIR spectra of unknown 

paint samples gained very similar results as obtained from experts.”[182] 

Another example of the application of statistics to analytical datasets acquired in 

paint examinations was reported by Latour et al.[183] In this study, diffuse 

reflectance spectra of pigment mixtures were simulated from a database of 

absorption and scattering coefficients of reference pigments typical of those used 

in works of art. 

Regarding additional reflectance studies, Poli and coworkers used mid-IR FORS 

to analyze a work of art without sampling.[184] A simple method for enhancing 

signals in sub-optimal conditions was successfully applied, and spectral 

corrections were evaluated for comparing the reflectance spectra with reference 

140

transmission spectra. Bacci and coauthors created a database of reflectance 

spectra in the NIR region and applied it in the analysis of a painting.[185] 

In their study, Vagnini et al. also examined FT-NIR, but for the non-invasive 

identification of natural polymers and resins.[186] Limitations of the technique 

included difficulty assigning bands, making it less-specific than mid-IR. Further, 

the technique is unable to distinguish single components in a mixture. Data 

treatment was found to be crucial. 

In two separate papers, Kendix et al. utilized far-infrared spectroscopy in the 

analysis of inorganic pigments.[187;188] FIR has the advantage when inorganic 

compounds are not active in the MIR. In this region of the electromagnetic 

spectrum, FIR technique also has an advantage over Raman when fluorescence 

interferes with Raman signals. 

Rizzo reported on using ATR-FTIR microscopy to analyze paint cross-sections, 

with complementary techniques still able to be applied to the same samples.[189] 

Historically, ATR on cross-sections has been difficult, but this study describes a 

crystal of optimized geometry and a viewing capability feature. 

Cotte and coauthors described synchrotron-based FTIR and reviews recent 

applications of the technique in the field of cultural heritage.[190] 

In their work, Delaney et al. concluded that shortwave IR and luminescence 

imaging spectroscopy in combination proved useful in identifying and mapping 

primary colorants in artwork, specifically one of Picasso’s paintings. When XRF 

was also utilized in site specific in situ analysis, assignment to individual 

pigments was further improved.[191] 

The use of mid-FTIR reflectance spectroscopy was studied in situ via fiber optic 

technology to monitor and evaluate the cleaning process applied to artwork in 

real time.[192] Results indicated that both inorganic and organic contaminants, in 

the form of calcium oxalate and a terpenic varnish thought to have been added to 

the painting at a later date as a protective coating, were successfully removed 

from the painting surface using the cleaning process employed. Results were 

verified via a benchtop FTIR system as well as through the use of GC/MS 

analysis of the cotton swabs used to remove the varnish coating and degradation 

products from the surface. Use of the reported in situ approach allowed for a 

non-invasive aspect to be added to the process of monitoring these critical 

cleaning treatments within the field of conservation and restoration. 

15. Raman Spectroscopy 

Over the past two decades, Raman analysis has increased significantly in its 

application to art and other cultural heritage. During this review period, several 

articles were published that include general information and reviews on 

141

application of the technique to works of art.[193-197] Further, the Journal of 

Raman Spectroscopy has a special issue entitled “Raman Spectrometry in Art 

and Archaeology III,” Volume 89, number 4, December 2007, pertinent articles 

from which have been included in this review. 

One focus of recent Raman studies was surface enhanced Raman spectroscopy 

(SERS). Chen and coauthors reviewed the technique to include the principle, 

instrumentation, and its use in art and archaeology.[198] Wustholz et al. studied 

the utility of SERS to analyze dyes. They highlight recent work and future 

challenges.[199] In their work, Whitney and coworkers applied the technique to 

red dyes and their mixtures, and Brosseau et al. reported the first SERS study of 

multiple artists’ materials, including organic pigments (both synthetic and 

natural).[200-202] Van Elslande and coauthors also applied the technique to 

organic pigments.[203] Lau et al. applied the technique in a different way by 

analyzing natural resins.[204] As opposed to normal Raman, SERS was able to 

overcome interference from fluorescence in extremely small natural resin 

samples. 

Multivariate statistics was also found to be useful for Raman studies. In two 

separate publications, Nevin et al. utilized Raman and PCA to analyze 

proteinaceous binding media, a material which is typically problematic for Raman 

analysis.[205;206] 

Organic pigments were the focus of multiple Raman publications during this 

review period. Lutzenburger and Stege noted that there are approximately 100 

synthetic organic pigments used in artists’ paints today.[207] A comprehensive 

and efficient sampling and analysis strategy was suggested, with attention to the 

practical strengths and limitations of the technique. Case studies were 

discussed, and for the organic pigments identified, the history of their use was 

also discussed. These authors collaborated with Schulte and others to study 

synthetic pigment identification by using both dispersive and FT-Raman on 23 

pigments.[208] Different excitation wavelengths were required to avoid 

fluorescence and heating of the artifacts. In many cases it was possible to 

identify both organic pigments and the main inorganic pigments or fillers. 

Discrimination power varied within different pigment classes and complementary 

chromatographic methods were recommended. Papliaka and coauthors used 

Raman and FTIR in combination to characterize the materials of a contemporary 

painting and to investigate the stability of synthetic pigments in a widely used 

contemporary binding medium for conservation purposes.[209] Red organic 

colorants were the focus of Schmidt’s and Trentelman’s study, and yellow 

organic pigments were analyzed by Ropret et al. and Colombini and Kaifas.[210- 

212] In the latter study, the authors also looked at orange organic pigments, as 

well as pigments found in automotive paints. 

Correia and coauthors took a different approach by examining inorganic 

pigments and extenders and, in combination with other analytical techniques, 

142

found Raman effective at establishing one artist’s palette.[213] Acquaviva, et al. 

utilized Raman as well as excimer laser techniques in order to study yellow 

pigments in egg yolk tempura.[214] Colomban also used Raman scattering to 

study colors produced on pottery and glass using metal nanoparticles.[215] 

Vandenabeele and coauthors compared the performance of five mobile Raman 

instruments through evaluation of spectral resolution, wavelength calibration, and 

the ability to focus through a varnish layer. The study has the potential to 

facilitate a better definition of the requirements for the design of a future mobile 

instrument.[216] Articles were also published that discussed case studies of 

Raman spectroscopy to answer questions about specific works of art (Hernanz 

2008, Baraldi 2009).[217;218] 

Both organic and inorganic pigments used in pre-mixed architectural paints from 

the first half of the twentieth century were discussed by Gautier, et al. with 

respect to their dual use by artists such as Picasso. Raman, XRF, and FTIR 

were used to analyze Ripolin ® , a commercially available brand of architectural 

paints which were manufactured in Europe in the late nineteenth century for 

architectural and marine applications. A major finding in this study was observed 

with Raman and confirmed by PLM, in that carbon black as opposed to ivory or 

bone black was used in the black Ripolin ® paint. Other findings included the 

observation that lead carbonate, as well as common pigments such as vermilion, 

cadmium red or yellow, emerald green, or cobalt violets, were not present in the 

Ripolin ® paint, whereas it is common in tubes of traditional artists’ paints. Lastly, 

the authors reported that these architectural paint formulations always create 

secondary and tertiary colors from combinations of primary colors rather than the 

addition of other mediums to achieve the desired effect.[219] 

Perez-Pueyo and coworkers in Spain described the issues encountered in 

Raman spectral interpretation due to the presence of intense fluorescence 

background signals. They proposed a morphology-based automated baseline 

correction and applied the concept to the analysis of artistic pigments. The 

authors concluded that this approach successfully removed fluorescence-related 

interferences while retaining the line shapes and positions of the Raman bands 

of interest.[220] 

In a separate study, Perez-Pueyo and coworkers identified artistic pigments 

using a fuzzy approach as applied to data acquired via Raman spectroscopy. 

The authors describe fuzzy logic as “a mathematical technique that follows the 

guidelines of the human reasoning mechanism for decision making with its ability 

to work from approximate data and find precise solutions.”[221] 

16. Fluorescence Spectroscopy 

Confocal microfluorescence spectroscopy was used by Claro et al. for the 

analysis of red lake pigments in binding media, and a database was 

developed.[222] The technique was subsequently applied to artwork 

143

samples.[223] Emission and excitation spectra were both acquired, and the 

results obtained were in agreement with information from other currently used 

techniques. 

Fluorescence excitation-emission spectroscopy (also called total emission 

spectroscopy) was employed by Nevin et al. for the differentiation of proteinbased 

binding media and in a subsequent study to evaluate the effect visible light 

and inorganic pigments have on the fluorescent properties of protein-based 

binding media.[224;225] In the former study, time-resolved fluorescence 

spectroscopy and imaging was also employed. In the latter, it was found that 

pigments can have a significant effect on the chemistry and observation of the 

fluorescence of binding media. Nevin et al. subsequently reported that 

synchronous fluorescence spectroscopy (SFS) could aid in the assessment of 

fluorescence as a function of pigment concentration.[226] 

In SFS, a fluorescence emission intensity spectrum is obtained by varying both 

the excitation and emission wavelengths simultaneously at a constant offset 

between excitation and emission monochromators. In 2008, Nevin and 

coauthors applied the technique in a novel way to nondestructively distinguish 

protein-based binding media. PCA was used, since small variations in spectra 

can correlate with particular chemical phenomena.[227] In 2009, Nevin et al. 

also used SFS with multivariate analysis on selected organic painting materials 

representing binding media and stated that the technique could also be used for 

analysis of colorants and dyes.[228] 

17. Terahertz spectroscopy 

A technique that is fairly new to the analysis of art is terahertz spectroscopy. In 

contrast to the IR region, the terahertz region (500 to 20 cm -1 ) provides 

information on the motions of entire molecules and functional groups. Fukunaga 

and coauthors developed a database of over 100 organic and inorganic 

pigments, natural resins, and synthetic polymers, and found that the technique 

can distinguish pigments, binders, and their mixtures.[229] This technique has 

been used to analyze organic binding media and to analyze hidden layers 

embedded in paintings. [230-232] 

18. X-Ray Based Techniques 

Many articles for the various X-Ray based techniques were also applications and 

case studies. These included SEM/EDS;[233;234] XRD;[235-239] XRF;[240- 

247] and combinations of X-Ray techniques.[248-251] 

For these techniques combined with Raman spectroscopy, refer to the following: 

SEM/EDS;[252] XRF;[253-258] and, both XRF and SEM/EDS.[259;260] 

144

XRD and Raman spectroscopy were combined by Brostoff et al. to identify 

synthetic organic pigments in powder samples, paint layer systems, and 

commercial artists’ paints. For the first time, XRD is demonstrated to be effective 

for the specific identification of synthetic organic pigment mixtures in acrylics and 

alkyds, but not in oil based paints.[261] 

Portable instrumentation was emphasized in multiple articles, most regarding 

portable XRF. Desnica et al. and Vasquez-Calvo and coauthors used the 

portable XRF device for screening purposes for subsequent laboratory 

analysis.[262;263] Further, Hocquet, Uhlir, and Buzanich and their respective 

coauthors designed new instruments and applied them in the field.[264-266] Abe 

and coauthors used portable XRF and portable XRD in conjunction with one 

another, and Giononcelli et al. introduced a newly designed combined portable 

XRF and portable XRD instrument.[267;268] The latter instrument demonstrated 

the possibility of identifying main crystalline phases in relatively short times. 

Bonizzoni and coauthors compared three different portable XRF systems in the 

characterization of pigments.[269] Van der Snickt and coauthors examined 

synthetic dyes and pigments in a large number of paintings by portable XRF in 

order to exploit the advantages of the technique, and Roldán applied portable 

XRF to pigments on rock art.[270;271] Finally, Bonizzoni et al. performed in situ 

analyses of paintings using XRF with visible reflectance spectroscopy.[272] 

PIXE was used by Grassi and coworkers to identify the elemental composition of 

a painting, and PIXE was combined with Rutherford Backscattering (RBS) by 

Salomon et al. to take advantages of the strengths of each technique.[273;274] 

PIXE is a non-destructive technique with high sensitivity down to trace element 

concentrations and can be implemented at atmospheric pressure; however, it is 

unable to provide depth information. RBS, on the other hand, can provide depth 

detail but cannot be readily implemented in air. Mendes and coauthors 

combined PIXE with Raman to study pigments and degradation products.[275] 

Lastly, Shaaban et al. used PIXE in combination with optical microscopy to study 

ancient Egyptian pigments.[276] 

Synchrotron-based techniques were also studied and reported on during this 

review period. However, due to a lack of availability to most practitioners, it will 

only be briefly mentioned here. D’Acapito et al. provided a technical overview of 

the instrumentation available on the GILDA beamline at the European 

Synchrotron Radiation Facility (ESRF), specifically X-Ray absorption 

spectroscopy and diffraction.[277] With regards to paintings, the beamline is 

used to study the degradation of pigments. XRD using synchrotron radiation was 

investigated by Sánchez del Río et al. and Welcomme et al., and XRF using 

synchrotron was reported on by Dik and coauthors and in two papers by Calza et 

al.[278-282] Some combination of the synchrotron versions of FTIR, X-ray 

absorption near-edge structure spectroscopy (XANES), XRD, and XRF were also 

used by several research groups.[283-289] 

145

Micro XRD analysis was discussed for pigment analysis by Nel and coauthors in 

proceedings from the AICCM National Conference in Brisbane, Australia in 2007. 

In this work, the authors observed that the technique can complement other nondestructive, 

non-invasive, and in-situ technologies, but was best applied to art 

and artifacts where the pigments were pure, mixing of pigments was minimal, or 

where the pigments within a mixture did not have significant differences in their 

absorption coefficients.[290] 

Van der Snickt and coworkers describe a combined micro-XRD and micro- 

Raman mobile device (PRAXIS) and compared the PRAXIS results to those 

obtained from synchrotron micro-XRD. The latter technique was required to 

show the presence of lead white in an illuminated manuscript. The authors also 

mention that despite all the information obtained from the aforementioned 

techniques, a supplementary organic method would also have been helpful in 

identifying some remaining unidentified components.[291] Herrera et al. did in 

fact combine multiple analytical techniques, including organic and inorganic 

analysis methods, in combination with synchrotron XRF and XRD, for the 

analysis of paintings.[292] A novel approach was taken by Ferreira and 

coauthors in the use of synchrotron X-Ray microtomography to obtain threedimensional 

data, thereby allowing for the reconstruction of sample surface 

morphology.[293] 

19. Nuclear Magnetic Resonance (NMR) 

One article was found during the review period that used NMR for paint analysis. 

In the article, Presciutti et al. used a compact and mobile NMR sensor to 

measure the thickness of paint layers in situ and to investigate binder aging. The 

work claimed to be the first step towards characterizing the three-dimensional 

morphology of different layers in paintings without contact.[294] 

20. Laser induced breakdown spectroscopy (LIBS) 

In 2007, Giakoumaki and coauthors published a review article outlining the basic 

physical principles and analytical features of LIBS, with a description of the 

instrumentation and technology, examples of LIBS use (including a section on 

pigments), and future prospects.[295] 

Osticioli and coauthors set out to determine the best working conditions for LIBS 

to remove the least amount of material during analysis while achieving the best 

quality in spectral response.[296] The optimized conditions were applied to a 

real painting, in which most of the inorganic pigments were identified. A 

significant limitation, however, was the alteration of pigments with regards to 

discoloration and darkening. This work was furthered in a study by the same 

lead author in which the effects of light exposure on egg white and egg yolk films 

was evaluated.[297] 

146

Natural versus artificial ultramarine blue pigments were differentiated by the 

presence of calcite in the former using LIBS, pulsed Raman spectroscopy, 

principal components analysis, and light microscopy in another paper by Osticioli, 

et al. [298] Several other combined LIBS and Raman studies were also carried 

out that demonstrated the efficiency of and complementary information resulting 

from the use of the techniques both in a single instrument setup [299-301] and as 

stand-alone technique.[302] 

21. Laser ablation inductively coupled plasma – mass spectrometry (LA- 

ICP-MS) 

LA-ICP-MS was reviewed by Giussani and coworkers in which the main issues 

relating to cultural heritage are discussed, followed by a description of the 

features, advantages, and drawbacks of LA-ICP-MS. Pigments are briefly 

mentioned.[303] 

LA-ICP-MS was combined with SEM/EDS and Raman by Resano et al. to 

characterize pigments in rock art. LA provided the most relevant data for 

associating samples and was capable of providing spatially resolved (both 

laterally and in depth) quantitative multielement information for elements present 

in a wide concentration range.[304] 

Arnold and coauthors used LA-ICP-MS for sourcing of materials and compared 

the results to instrumental neutron activation analysis (INAA). Similar data was 

obtained but LA-ICP-MS was preferred due to its ease, smaller analytical 

investment, and near non-destructiveness. LA also determined some elements 

that INAA could not.[305] 

22. Gas Chromatography 

GC was used in combination with different sample preparation techniques and 

detectors for the analysis of various organic components used in cultural 

heritage. 

Derivatizing reagents and GC/MS were used by Caruso et al. for the analysis of 

the varnish (fatty acids) and the glue (proteinaceous amino acids) on a musical 

instrument and also by Echard and coauthors for the analysis of varnishes on 

three ancient musical instruments.[306;307] For a general review of the 

characterization of varnishes on musical instruments, refer to [286]. Edwards 

and coauthors used Raman spectroscopy to pinpoint areas for GC/MS analysis 

of varnishes on ancient Egyptian sarcophagal fragments, while Singer and 

McGuigan used GC/MS to analyze fatty acids, amino acids, and natural resins in 

paints.[308;309] Gautier and Colombini attempted to determine the 

proteinaceous binder composition in wall paintings by purifying proteins through 

the removal of inorganic salts that impair the determination of the protein via 

amino acid analysis.[310] 

147

Bonaduce and coauthors used derivatization and GC/MS for the analysis of 

sugars in plant gums in mural paintings. The result was one peak for each 

compound, providing simple and highly reproducible chromatograms. A decision 

scheme for gum identification and the PCA analysis of the relative sugar 

percentage contents of reference samples were employed in the identification of 

unknowns.[311] 

Sutherland discussed products formed from reactions of alcohols with a 

quaternary ammonium derivatization reagent to aid in interpreting data obtained 

through the use of that reagent. Although the formation of multiple reaction 

products is undesirable, the reagent has the advantage of giving very 

characteristic fragmentation patterns with EI MS.[312] 

Pyrolysis GC/MS was applied by Frade and coauthors to analyze lacquers from 

Asia. Py-GC/MS was previously established to be the only technique capable of 

distinguishing lacquers, films of which are insoluble and have complex structures. 

This article represented a new method to identify the three kinds of Asian 

lacquers using their characteristic m/z 104 and 108 mass chromatogram 

profiles.[313] 

Both derivatization and pyrolysis were used with GC/MS in several additional 

studies. Doménech-Carbó et al. analyzed PVA resins in two separate studies. 

The first used online silylation Py-GC/MS to characterize the resins, and the new 

method led to the unambiguous identification of PVA. In this study, the authors 

stated that they thought repeatability was improved as compared to conventional 

Py-GC/MS.[314] In the second study, Doménech-Carbó evaluated the 

plasticizers in PVA emulsions as potential carbon sources of 

microorganisms.[315] Silva and coauthors analyzed PVA paints for plasticizer 

content and evaluated the additives (surfactants, coalescing agents, defoamers, 

freeze-thaw agents, and thickeners).[316;317] In their work, He Ling et al. 

analyzed Chinese binding media to gain knowledge on the materials used in 

Chinese artworks. Fingerprint ions for the various types were determined.[318] 

Ketone resins used as varnishes were unambiguously identified by Doménech- 

Carbó and coauthors. Compared to conventional Py-GC/MS, the online silylation 

Py-GC/MS method increased structural information obtained for the materials 

studied.[319;320] 

Another study by Menke and coauthors applied direct temperature-resolved 

mass spectrometry (DTMS) to the detection of four different organic pigment 

classes: azo, quinacridone, dioxazine, and phthalocynanine in a variety of 35 

reference samples as well as an artist’s specific paint samples. The authors 

concluded that the technique provided the best results when used in the negative 

ion mode using chemical ionization for the majority of the synthetic organic 

pigments that were studied. They discussed that the limited fragmentation 

provided a stronger and more readily detectable molecular ion pattern which 

148

could be distinguished from lower mass-to-charge (m/z) species typically 

generated from other constituents in paint formulations.[321] Lastly, Kirby et al. 

reported on the use of laser desorption mass spectrometry for synthetic organic 

pigments as employed in works of art.[322] 

23. Capillary Electrophoresis (CE) 

Compared to GC, CE requires simpler sample pretreatment (does not require 

derivatization) but the separation efficiency and detection sensitivity is not as 

good. Two separate studies describe these differences. To demonstrate how 

CE can be used successfully, Kaml and Kenndler described using CE for the 

analysis of natural organic binding media.[323] Findeisen and coauthors 

analyzed diterpenoic acids used in coatings and varnishes.[324] 

24. Liquid Chromatography (LC) 

The final separation technique applied to paint analysis was liquid 

chromatography. Checa-Moreno et al. used HPLC with a diode array detector 

(DAD) to identify proteinaceous binders. In such an application, traditional 

means of identification are prone to difficulty. Therefore, SIMCA (soft 

independent modeling of class analogy) was evaluated and compared to the 

more traditional strategies.[325] In their research, Leo and coauthors used nano 

LC-MS/MS with ESI to further apply proteomic strategies to the identification of 

proteins in paintings.[326] 

25. Matrix-assisted laser desorption ionization - time of flight mass 

spectrometry (MALDI-TOF) 

Researchers have demonstrated that MALDI-TOF can be utilized in paint 

analyses. Kuckova and coauthors identified proteinaceous binders in very small 

samples and discussed how MALDI-TOF can distinguish among main binder 

classes, is a relative speedy technique, and has low cost following the purchase 

of the MS.[327] Soltzberg et al. used the technique in both positive and negative 

modes to characterize dyes and pigments. The technique provided a convenient 

and versatile method for such a purpose.[328] 

In two separate publications, Hoogland and Boon combined MALDI with nano- 

ESI-MS to analyze poly(ethylene glycol) additives in acrylic artists’ paints. In the 

Method development and application of the technique to paints in described in 

the two papers. In some instances, the authors assert that it was possible to 

suggest a specific brand of paint.[329;330] 

149

26. Infusion Mass Spectrometry 

A final mass spectrometry technique investigated was infusion mass 

spectrometry.[331] This technique involves simple hydrolysis and extraction as a 

sample preparation technique that does not involve a derivatization step, 

followed by direct introduction to the MS. In this artwork application, drying oils, 

used as binding media and varnishes, were characterized. 

27. Imaging (Tomography) 

Optical Coherence Tomography (OCT) is a technique for obtaining sub-surface 

images of translucent or opaque materials at a resolution equivalent to a lowpower 

microscope. It is similar to ultrasound and can penetrate up to several 

millimeters to provide cross-sectional images. Targowski et al. provide an 

overview of the technique and describes an instrument especially designed for 

art diagnostics.[332] Liang and coauthors report progress on optimizing and 

designing an OCT instrument for in situ use in museums.[333] Adler et al. 

discuss the technique further and claim that it is well suited for analyzing artwork 

where a surface layer obscures details of interest.[334] In two separate papers, 

Latour and coauthors use the technique to obtain pigment information and 

demonstrate how the technique can be applied to wood varnishes.[335;336] 

Computed Tomography (CT), another imaging technique traditionally used in 

medical diagnostics, was shown to be applicable to artwork analysis by providing 

morphological and physical information on the inner structure of artworks.[337] 

Its use with radiography was discussed. 

28. Determination of Materials and Artistic Techniques 

A variety of the techniques described thus far were used to evaluate the 

materials and/or techniques of various artists, collections, or geographic 

locations.[338-344] Furthermore, a variety of substrates were analyzed, 

including alabaster[345], archaeological pastes[346], drawings and illuminated 

manuscripts [347;348], pottery or stoneware[349-352], religious items[353;354], 

rocks[355], tiles [356], and walls[357-364]. 

29. Conservation, Degradation and Forgery Detection 

A range of the aforementioned techniques were also used in various 

combinations in order to study degradation/aging processes in painting 

materials.[297;365-369] Since this has little practical application for forensic 

paint analyses, the appropriate references are merely listed. For a thorough 

discussion on the various techniques available for materials conservation, 

including paints, consult Analytical Techniques in Materials Conservation.[370] 

Further, Applied Physics A published a special issue entitled “Science and 

Technology of Cultural Heritage Materials: Art Conservation and Restoration,” 

Volume 92, number 1, July 2008 that may be of interest. 

150

A review by Dawson discusses the basic materials used in the creation of art as 

well as how these components degrade.[371] This article provides an excellent 

foundation for understanding conservation and restoration. For a comprehensive 

review of the most recent advances in the identification and determination of 

organic compounds present in art and art conservation materials, consult 

Doménech-Carbó’s article.[365] 

Laser technology, specifically, appears to be well established for conservation of 

cultural heritage. In his book, Lasers in preservation of cultural heritage, Fotakis 

et al. describe the principles and applications of lasers in cultural heritage.[372] 

Further, several journal articles provide similar information.[373-376] 

Ultraviolet fluorescence spectroscopy was reported on by Thoury et al., where 

varnishes were studied in situ on prepared samples as well as works of art.[377] 

The process of teaching scientists and non-science conservators how to conduct 

non-destructive, low-cost analyses was described by Brasuel and coauthors. 

Using three different experimental models, the authors describe how students 

use reflectance and UV-vis spectroscopies to detect forgeries in artwork and 

estimate year of origin. They are also introduced to multivariate calibration, and 

assessment of the organic components of mixtures versus pure materials via Py- 

GC/MS data acquisition.[378] 

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[340] Daniilia S, Andrikopoulos K, Sotiropoulou S, Karapanagiotis I. Analytical 

study into El Greco's baptism of Christ: clues to the genius of his palette. 


[341] Gil M, Carvalho M, Seruya A, Ribeiro I, Queralt I, Candeias A, et al. 

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[343] Appolonia L, Vaudan D, Chatel V, Aceto A, Mirti P. Combined use of 

FORS, XRF and Raman spectroscopy in the study of mural paintings in the 

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[344] Karapanagiotis I, Minopoulou E, Valianou L, Daniilia S, Chryssoulakis Y. 

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iconography. Analytical Chemistry 2009; 647:231-242. 

[345] Pérez-Arantegui J, Ribechini E, Pardos C, Colombini M. Chemical 

investigation on black pigments in the carved decoration of sixteenth 

century alabaster tombs from Zaragoza (Spain). Analytical and 


[346] Maier M, de Faria D, Boschín M, Parera S, Bernal M. Combined use of 

vibrational spectroscopy and GC-MS methods in the characterization of 

archaeological pastes from Patagonia. Vibrational Spectroscopy 2007; 

44:182-186. 

[347] Bruni S, Caglio S, Guglielmi V, Poldi G. The joined use of n.i. spectroscopic 

analysis-FTIR, Raman, visible reflectance spectrometry and EDXRF to 

study drawings and illuminated manuscripts. Applied Physics A: Materials 

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[348] Trojek T, Čechák T, Musílek L. Recognition of pigment layers in illuminated 

manuscripts by means of Ka/Kb and La/Lb ratios of characteristic X-rays. 

Applied Radiation and Isotopes 2010; 68:871-874. 

[349] Mangone A, Giannossa L, Colafemmina G, Laviano R, Traini A. Use of 

various spectroscopy techniques to investigate raw materials and define 

processes in the overpainting of Apulian red figured pottery (4th century 

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situ non-invasive investigation on the painting techniques of early Meissen 

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powerful means to investigate the glazed coating of ancient ceramics. 

Journal of Nano Research 2009; 8:141-146. 

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'in situ", characterization of pigments in Minoan pottery at the 

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39:230-232. 

[353] Vahur S, Sibul K, Ehasalu P, Sammelselg V, Leito I. Paint investigation of 

altar and pulpit in the church of St. Mary, Vigala, Estonia. Preservation 

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[354] Vornicu N, Geba M, Bibire C, Non-destructive analysis in a study of the 

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FT-IR and GC-MS information for identification of inorganic and organic 

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This is the FBI Laboratory’s publication number 10-27. 

Names of commercial manufacturers are provided for identification purposes 

only, and inclusion does not imply endorsement of the manufacturer, or its 

products or services by the FBI. The views expressed are those of the authors’ 

and do not necessarily reflect the official policy or position of the FBI or the US 

Government. 

181

Examination of Fibres and Textiles 


Ray Palmer 

Principal Forensic Scientist (Fibres) 

The Forensic Science Service Ltd, UK 

Email: ray.palmer@fss.pnn.police.uk 

182

  


1. INTRODUCTION  184  

2. GENERAL  184  

3. CASE REPORTS  185  

4. TEXTILE/ FIBRE DAMAGE  189  

5. EVIDENCE COLLECTION  190  

6. SIGNIFICANCE OF EVIDENCE  190  

7. ANALYSIS  192  

8. THE TEXTILE INDUSTRY  196  

9. THE FUTURE  196  

10. SUMMARY  196  

11. REFERENCES  197  

183


This report catalogues the research, advances and application of scientific 

methodology relating to the forensic examination of fibres since the 15 th 

INTERPOL Forensic Science Symposium held in October 2007. This report 

primarily consists of a literature review of published articles in forensic science 

journals and the proceedings of various working groups between May 2007 

and July 2010. It also contains references from other sources such as the 

internet. This report should therefore be considered a ‘follow-on’ from that 

produced by the author [1] in 2007. 

2. General 

The European Fibre Group (EFG) of the European Network of Forensic 

Science Institutes (ENFSI) and the Fibre subgroup of the Scientific Working 

Group for Materials Analysis (SWGMAT) led by the FBI, continue to be the 

main organisations promoting and developing this evidence type worldwide. 

Many of the citations in this document originate from the activities of these 

groups and/ or their members. 

The EFG remains committed to disseminating best practice and over the last 

3 years has continued its program of themed ‘development’ workshops, aimed 

at court reporting practitioners with less than 3 years experience. The 2008 

workshop was held at the ‘Spiritka’ facility in Prague, where the topics 

covered were identification, comparison and interpretation. The next 

workshop is planned for spring 2011, with the theme to be decided. 

The planned production by the EFG of a document outlining the best use of 

fibre evidence in terrorist cases is now complete and the official launch of the 

“Fibre and Textile Evidence in Terrorist Cases - A guide for investigators.” will 

take place at the 16th International Forensic Science Symposium, ICPO- 

Interpol General Secretariat, Lyon, 2010. This document is not intended as a 

proscriptive laboratory or scenes of crime ‘manual’, but rather as a source of 

reference for investigators of the ways in which the forensic examination of 

fibres and textile materials can be applied to specific areas in the 

investigations of terrorist casework, counter-terrorism initiatives and other 

major crime investigations. 

The information contained in this document represents a pan-European 

collaboration of operational practitioners in this evidence type in direct 

response to the challenges presently faced by law enforcement agencies 

throughout the world. In addition, the revisions to the EFG Fibre Examination 

Guidelines document (originally published in 1998) is almost complete. 

Similarly, in 2008, the SWGMAT Fibre Sub-Group worked on revising the 

Fibre Analysis Guideline, and began to put together an admissibility package. 

The admissibility package will discuss the underlying science behind fibre 

analysis, with the goal of providing a contained summary used to explain in 

courts of law what forensic fibre analysis is. In 2009, SWGMAT published an 

Expert Reporting Guideline. This document describes the minimum elements 

184

that should be included in a forensic report of examination to satisfy the 

requirements of the American Society of Crime Laboratory Directors 

Laboratory Accreditation Board (ASCLD/LAB)-International – Supplemental 

Requirements, ISO/IEC International Standard 17025, and United States 

Federal Rules of Evidence. In 2010, the Fibre Sub-Group finalised changes 

on four chapters of the Fibre Analysis Guideline and submitted them to 

SWGMAT for voting. These documents were approved and should be 

published shortly. The Fibre Sub-Group is continuing to consider admissibility 

guidance, and has begun discussion of data interpretation. 

Colleagues in the EFG and SWGMAT continue to collaborate and 

representatives from each organisation attend the annual meeting of teach 

group. A summary of the background to the formation of the EFG and other 

ENFSI working groups is provided by Becker [2] 

The recent report of the US National Academy Committee on identifying the 

needs of the forensic science community [3] highlighted the need for better 

funding and a more unified plan for developing forensic science research in 

the US. Whilst there is a temptation to regard the conclusions of this report as 

irrelevant from a European perspective, the fact remains that many members 

of European working groups are finding it harder to plan, fund and carry out 

research due to the present economic climate and the increasing demands on 

their time. Despite this, over the past three years our knowledge base 

continues to advance, and in particular (see ‘case studies’), it is gratifying to 

see the added value that fibre evidence has provided in ‘holistic’ approaches 

to major complex enquires. The need to continue the promotion of the ‘added 

value’ of fibres (and other trace evidence) has been well argued by Robertson 

and Roux [4]. 

3. Case reports 

Over the past three years there have been numerous instances where fibre 

evidence has proved crucial in the investigation of complex major inquiries [5- 

18], very often in providing an investigative steer through the provision of 

intelligence and/ or added value to other evidence types employed. 

Through the advent of TV shows such as ‘CSI’, criminals (and the population 

generally) are much more aware of the potential evidence which may be 

gathered from a crime scene. This so-called ‘CSI effect’ has resulted in many 

instances where those involved in the commission of a crime have taken 

active steps to remove trace evidence relating to their activities. Hutchinson 

[5] reported on a case where the victim of a homicide had been transported to 

a deposition site in a car belonging to one of the perpetrators. The 

perpetrators then donned personal protective equipment as used by scenes of 

crime personnel, and used UV light and tape lift techniques (as seen on TV) in 

an attempt to remove any potential trace evidence from the victim. Despite 

their attempts, significant fibre evidence linking the deceased to the vehicle 

was ultimately recovered. 

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The following case studies [6-8] show that (contrary to expectation) significant 

fibre evidence can be recovered from victims of homicide deposited in an 

aquatic environment and that this can be used as a powerful intelligence tool 

providing an investigative steer to the investigative team; 

Palmer [6] detailed the circumstances of the ‘Suffolk Serial Killings’, when the 

naked bodies of five young women were discovered in various locations on 

the outskirts of the town of Ipswich in England in December 2006. Two of 

these women were recovered from a river after immersion for a number of 

weeks, whilst the remaining three had been deposited in woodland and 

exposed to a combination of heavy rainfall and wind. Despite the adverse 

conditions to which the bodies of these women had been exposed, 

combinations of fibre collectives common to all of the victims were identified in 

debris recovered from their bodies. Fibres were recovered from the hair of the 

two women immersed in water, using a modified palynological technique. 

Using fibres as an intelligence tool, sources for these fibre collectives were 

eventually identified within the home environment of a suspect, Steve 

WRIGHT, who was subsequently found guilty of all five murders. 

Sulkava [7] described a double homicide where there was a five year gap 

between recovery of the victims who had both been deposited in an aquatic 

environment (the first in the sea, the second in a lake). Using a combination 

tape lifting, vacuuming, combing and washing and filtration of debris of the 

clothing of the second victim, fibre and other trace evidence was recovered 

which was to prove crucial in the conviction of the suspect. 

De Wael [8] reported on a case involving the abduction and murder of two 

young girls, whose bodies were found in a drainage canal three weeks after 

their disappearance and subjected to fast flowing water during this period. 

The address of the prime suspect in the case was searched and a number of 

items of clothing interest were recovered, however, these had been washed. 

Despite this, painstaking examinations were not only able to demonstrate a 

two way primary transfer between the victims and the suspect, but also a 

secondary transfer of fibres to the victims, from the duvet covers belonging to 

the girlfriend of the suspect, via the suspect himself. The majority of fibres 

were recovered from ‘concealed’ areas of the garments in question (e.g. 

seams on inside surfaces). At trial, the suspect was found guilty and 

sentenced to life imprisonment. 

Shaw [9] presented a case study on a series of rapes which occurred over a 

three year period in the state of Illinois, USA. In each case, the perpetrator 

entered the home of a single woman, bound the victim and after sexually 

assaulting them, forced them to bathe. The head and wrists of one of the 

victims were bound with duct tape. Examination of the duct tape revealed the 

presence of two distinct fibre collectives, however, nothing further of 

significance was found on any of the other victims. Following a report of an 

attempted break- in, the suspect, a policeman, was arrested. A balaclava and 

jacket found in the suspects home were found to be sources for the fibre 

collectives on the duct tape used to bind one of the victims. The suspect was 

subsequently found guilty at trial. 

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Palmer [10] described a case where various parts of a dismembered body 

had been discovered in countryside over a large geographical area. The 

majority of these body parts had been placed in plastic sacks and wrapped in 

duct tape. Analysis of the adhesive side of the duct tapes revealed a collective 

of blue flock fibres, as well as green polyester and cotton fibres common to 

each of the body part wrappings. Having provided the investigative team 

intelligence in terms of the likely sources for the collectives, sources were 

found in the victims home – indicating this was where the body parts had 

been wrapped. This address was now occupied by a business associate, who 

had been claiming to be the deceased in order to gain access to his assets. At 

trial, the suspect pled guilty to murder. 

A presentation by Bennett et al [11] described the investigation of a murder 

case in which the only trace evidence found as a result of the examination of 

the scene, was a number of coarse, dark fibres which were stuck to the soles 

of the victim's shoes. Nine of the fibres were grey polypropylene, 12 were blue 

polypropylene, and 50 were black polyester. These fibres were found to 

originate from the carpet of the suspect's vehicle, with almost all other 

possible sources of the fibres being eliminated. At trial, the main issue was 

the persistence of the fibres on the soles of the victim's shoes. Tests were 

carried out in order to investigate the factors affecting the transfer and 

persistence of carpet fibres to shoe soles. The case illustrates not only how 

data from transfer and persistence studies on shoes formed a vital part of the 

prosecution's case, but also the importance of industrial enquiries in these 

circumstances. 

Schwetz [12] presented the details of a case involving the abduction and 

murder of a young girl in 1987. The prime suspect in this case was released 

without charge. 20 years later the case was re-opened and a number of fibre 

collectives were identified on the victims clothing. A search was carried out in 

the suspects environment a sources for the collectives were found on blanket 

and bed linen found in the suspects flat. In addition, fibres matching those of 

the victims anorak were found. At trial, the suspect was subsequently found 

guilty. This case exemplifies that meaningful evidence can be obtained even 

where there are significant periods of time between the offence taking place 

and an examination being carried out. It also exemplifies the intelligence value 

of fibres in identifying places and items of interest. 

Krauss et al [13] reported on an investigation of a case in which a woman was 

found hanging in her garage resulted in the medical examiner concluding that 

the manner of death was suicide. However, tape lifts from the victim's hands 

and clothing, and from the scene and other locations were subsequently 

examined for fibre evidence. Based on the discovery and examination of 

greyish-blue polyester fibres and grey viscose fibres recovered from the 

scene, it was concluded that the victim had come into contact with a garment 

from an unknown person. A match was found with corresponding fibres from 

an item of clothing found in the home of the victim's estranged husband. This 

evidence resulted in this individual's conviction for the murder of his estranged 

wife. 

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Suzuki et al [14] described the investigation of a murder case the suspect 

stated that he had found the victim's body when he broke into the crime scene 

in order to commit theft. Consequently, the crime scene was reconstructed in 

accordance with the suspect's confession, and the suspect's behaviour was 

simulated according to the confession. In order to assess the reliability of the 

suspect's statement, the distribution and number of characteristic fibres 

collected in the simulations and during the actual investigation were compared 

which revealed a significant difference in the numbers of fibres found at each 

area of the victim's body, the mattress, and the blankets. The difference in the 

number of fibres found close to the victim's head was particularly significant. 

The findings were presented as evidence in court and the defendant was 

subsequently found guilty of murder. 

Sulkava [15] reported on a murder case where unusually the target fibres in 

question, were blue denim. The victim in this case was a hermit who 

possessed no clothing containing fibres of this type. The unusual 

circumstances of this case are discussed as is the role of the fibre evidence in 

combination with other evidence types in securing a conviction. The case 

illustrates the context sensitivity of forensic casework and how a fibre type 

normally considered of no evidential value proved crucial in the investigation. 

It is interesting and gratifying to note that in each of the above cases of 

homicide, the fibre evidence was used early on in the investigation as an 

intelligence tool - exemplifying their value as a primary application tool, rather 

than a ‘last ditch’ possibility (as is often perceived by some investigators). 

Fibre evidence continues to be used routinely in the investigation of a number 

of cases involving motor vehicles, where there is dubiety over who was driving 

the vehicle in question. In order to correctly identify who was driving at the 

time of a traffic accident or vehicle theft, as many seats as possible from the 

vehicle(s) should be examined for fibre traces. 

Majdic [16] described a case illustrating the importance of determining the 

relative distribution of fibres from target garments on each of the seats of a 

vehicle. The case clearly demonstrates it is not sufficient to consider only the 

fibre findings from the driver's seat in order to answer the specific question 

and emphasises the problems encountered through poor submission strategy 

resulting from attempts to cut costs/ corners. 

Whilst the distribution of fibres transferred to vehicle seats can be helpful in 

identifying the putative driver, the use of examinations for fibre plastic fusions 

remains the method of choice for this type of investigation. Schaich [17] and 

Irwin [18] presented case studies exemplifying the power of this approach in 

unequivocally identifying the driver of a vehicle at the time of the collision. The 

presentation by Irwin [18] was particularly interesting, in that it illustrated how 

approaches using blood distribution in these circumstances can produce the 

wrong result. 

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4. Textile/ fibre damage 

Was-Gubala [19] studied the colour changes in several types of textiles due to 

the long-term effects of exposure to laundry detergents. A 14-day study was 

carried out using blue, red, and grey/black cotton, wool, acrylic and polyester 

textiles. Colour changes were assessed visually by comparing the textile 

samples with the untreated (control) material. Fluorescence microscopy (UV 

excitation filter) was used to monitor the kinetics of the changes in the colour 

of the fibres. The changes observed are assessed from the perspective of 

both a fibre analysis expert and the average user/consumer of the products 

involved. 

Dillinger [20] presented the results of a study to determine the damage 

characteristics of dog bites on clothing. This study formed an important part of 

the prosecution case in the investigation of a homicide involving a sword. 

A presentation by Bennett [21] advocated a more holistic approach to fabric 

damage examination and presented a number of case examples in Australia 

where they had found this approach to be beneficial. 

Canetta et al [22] assessed the application of atomic force microscopy (AFM) 

to the examination of nanoscopic morphological changes in the surfaces of 

fabrics for the first time. The study material consisted of cotton, woollen, and 

viscose textile fibres exposed to burial in different types of soil and immersion 

in water (sea and freshwater) for various lengths of time. Examination of the 

resulting AFM images allowed the surface texture parameters of the 

environmentally stressed fabrics to be measured as a function of the 

exposure time. The finest details of the fabric surfaces could also be 

visualised, allowing the detrimental effects of the imposed environmental 

conditions to be distinguished. This work shows that AFM is potentially a very 

useful tool for the forensic examination of textile fibres for distinguishing 

between different environmental exposures and forced damage to fibres. 

De Wael, and Van Vaerenbergh [23] described a case where the charred 

remains of a Molotov cocktail were submitted to the laboratory for 

examination. The fibres and textiles laboratory was required to examine the 

textile remnants in order to determine the original colour and characteristics of 

the material. In this paper they demonstrate how a combination of 

microscopy, microspectrophotometry and hot-stage microscopy allowed 

examiners to determine the original colour of the fabric and the type of fabric 

involved. 

Hess [24] reported on a case where it was required to determine whether an 

injured party had attempted self immolation or was the victim of an accident. 

The various factors affecting the combustion of textiles, including propagation 

rates are discussed. These factors, taken into consideration along with testing 

of garments and solvents in question, enabled the investigation to conclude 

that the injuries were self inflicted. 

189

Many of these cases have produced learning points for future investigations 

and/ or been the drivers for further research. 

5. Evidence collection 

De Wael et al [25] compared five commercially available adhesive tapes in 

terms of fibre uptake and saturation, recovery, and ease of analysis. Based on 

the results obtained, a high-tack adhesive tape was selected for forensic fibre 

sampling two different micro trace kits, the first of which is mainly used for the 

sampling of vehicles and the second of which is a 1:1 tape lifting kit for the 

recovery of fibres from cadavers. 

Whilst tape lifting remains the main method of fibre recovery, Allen et al [26] 

described the use of a commercially available plastic rod with electrostatic 

properties, which can be used to recover fibres from surfaces inaccessible to/ 

or unsuitable for tape lifts (e.g. knife clasps, gun barrel, plastic bags). The 

electrostatic properties of the rods, and the recovery rate are assessed in this 

study. 

6. Significance of evidence 

Whilst there can be no doubt that the development of new analytical 

techniques and methodologies are an extremely important aspect of fibre 

evidence, even the most sensitive discriminating analytical technique is 

rendered ineffective if its results cannot be applied to answer specific case 

work related questions. In Europe at least, simply reporting the results of fibre 

analysis in terms of ‘match’ / ‘non –match’ is becoming less and less 

acceptable and there is now the requirement to place these within the context 

of the circumstances peculiar to a specific case. 

Over the last three years, work has continued to provide data which assists 

the practitioner (and the courts) in evaluating the results of analysis. Much of 

this work has been driven through questions posed in operational casework. 

Palmer and Burch [27] performed a combined investigation into the transfer 

and retention properties and fibre population study of the skin of living 

subjects. Fibres were transferred to the bare arms of living subjects and their 

persistence determined at intervals up to 24h, during which normal daily 

activity was undertaken. Decay curves showed an initial rapid loss followed by 

an apparently exponential decay. No target fibres remained after 24 h. The 

length distribution showed a shift towards shorter fibre lengths and the 

differential shedding results for a polyester/cotton mixture showed a small 

bias towards the retention of cotton. The population of coloured fibres on bare 

skin was classified according to perceived colour, length, generic class and 

presence or absence of delustrant. The study indicated that perception of skin 

as a non-retentive surface may be over simplistic. 

A study by Chewning et al [28] was undertaken to examine the persistence of 

fibres on the interior and exterior surfaces of ski masks during transportation 

to the FBI Laboratory and during evidence processing in order to assess the 

190

case for separate examinations of the interior and exterior of a mask. To this 

end, ski masks (n = 20) were each seeded with 50 test fibres on either the 

interior or exterior only. The masks were subsequently packaged, shipped, 

and processed according to standard procedures before the final recovery 

locations of the fibres were documented. The results obtained suggested that 

11 (55%) of the ski masks showed evidence of test-fibre transfer redistribution 

at some point during the study, although the number of fibres transferred only 

ranged from one to three. The study concluded that the probability that a fibre 

will be recovered from the same side on which it was deposited is higher than 

the probability that it will be recovered from the opposite side, but an examiner 

cannot conclude with certainty that fibres recovered in the laboratory from the 

interior or exterior of an item of evidence were originally deposited on that 

side. In many cases, processing of all the surfaces together may be 

acceptable. 

In another transfer and persistence study, De Wael and Gason [29] 

investigated the properties of microfibres as relatively little research has been 

carried out into this aspect of these fibres and further data was required in 

order to assist the interpretation of fibre findings associated with a murder 

case. The results obtained show that large amounts of microfibres can be 

transferred, although the degree of transference depends on the nature of the 

contact and the structure of the microfibre garment. 

Coyle and Jones [30] surveyed the use of flock in the interior of a number of 

motor vehicles. All vehicles had samples of flocked material in their interiors. 

Flocked material was found on the window channels, window frames, door 

edges and in glove compartments or central console compartments. The front 

seats of these vehicles were examined for loose flock fibres. All vehicles had 

flock fibres present on the front seats. In the majority of vehicles, flock 

originating from material in the vehicle's interior was present on the front seats 

of the vehicle. The number of flock fibres present on the front seats varied 

greatly from car to car but did not appear to be dependant on the type of seat 

fabric or on which areas in the car were flocked. The potential for these to be 

transferred to an occupant and the importance of taping the clothing of a 

suspect, where a link to a vehicle is in question, is discussed. In a follow up 

study [31], the authors performed a target fibre study to determine how likely it 

is that a particular flock fibres from a vehicle is likely to be encountered 

through chance. As with previous studies using other fibre targets, the authors 

concluded an adventitious match was highly unlikely. 

Dillinger [32] used a casework example is used to illustrate the investigative 

benefits of examining textile labels on items of clothing in order to obtain 

useful information with regard to clothing evidence. Such labels allow the 

manufacturer to provide exact details regarding the production and sales time 

period of the garments to which they were fitted. Also, the examination of 

textile labels allows investigators to reach conclusions relating to the 

authenticity of clothing, production figures, year of production, and possibly, 

the point of sale. Dillinger [33] also reported on new technical aspects relating 

to video imaging of textile materials and demonstrated how various factors 

associated with this method could lead to erroneous conclusions regarding 

191

the nature of the garment in question being drawn. The need for caution in its 

application and an appreciation of the factors affecting the technology are 

emphasised in intelligence led casework. 

7. Analysis 

Due to the prevailing economic climate, now more than ever purchase of new 

instrumentation and adoption of new methods of analysis are dependent upon 

demonstrable improvements in time, resources and/ or increased 

discrimination before uptake is justified in terms of cost. Nevertheless, over 

the past three years research in this area has continued. 

Biermann [34] carried out a study to determine the variation in blue and red 

cotton fibres using light and fluorescence microscopy, UV/Vis 

microspectrophotometry with particular attention being paid to the recurrence 

of certain spectral patterns. The importance of spectral information in the UV 

range is re-emphasised, as is the importance of colour in the forensic 

comparison of cotton fibres. The author demonstrated that the combination of 

tests employed can give a high power of discrimination, (DP c. 0.98) 

enhancing the evidential value of red and blue cotton fibres in fibre transfer 

cases. 

Similar results were obtained in a study of non-denim blue cotton fibres by 

Palmer et al [35] to determine the degree of discrimination obtained between 

non-denim blue cotton fibres using visible-UV range MSP alone. The 

discriminating power of visible range MSP was calculated to be 0.89 for 'midblue' 

garments and 0.87 for 'dark blue' garments. Extending MSP into the UV 

range increased discrimination by 7%, giving a discriminating power of 0.96 

for both mid and dark blue cotton fibres, which was similar to that reported by 

Biermann [34]. The implications of this study for casework are discussed and 

a revised analytical pathway for the comparison of this fibre type/colour 

combination using MSP as a primary screening tool is proposed. 

Research into the application of Raman spectroscopy to the forensic 

examination of textile fibres has continued since the last review, with 

members of the European Fibre Group being the most active in this area [36- 

38]. Despite this research, adoption of this technique into the majority of 

operational laboratories is still poor, possibly because it has still to 

demonstrate substantial advantages over the combination of existing 

techniques and in many respects has been shown to be complimentary; 

Massonett and Buzzini [36] compared Raman spectroscopy against 

microscopy, UV-Vis MSP and TLC in the discrimination of acrylic, cotton and 

wool fibres. The results of their study showed that in the majority of cases 

microscopy and UV Vis MSP provided the greatest discrimination and that the 

optimal analytical sequence varies with the fibre type/ colour combination 

(according to the discrimination power). 

Coyle et al [37] investigated the use of Raman spectroscopy in the subclassification 

of cellulosic fibres. Whilst it was possible to distinguish the 

192

polymeric backbone of colourless fibres, the dye present in coloured fibres of 

this type obscured this detail, rendering the technique unsuitable for this 

purpose in the vast majority of cases. 

Lepot et al [38] outlined five casework examples in which Raman 

spectroscopy was found to be a good complementary method to 

microspectrophotometry (MSP). They found that whilst absorption spectra in 

the visible range may occasionally be characteristic of a certain dye, this 

could be subsequently identified unambiguously by Raman spectroscopy 

using a spectral library. In other cases, comparing the Raman spectra of 

reference fibres and suspect fibres resulted in an improvement in the power of 

discrimination. It was concluded that Raman spectroscopy is a powerful 

method applicable to routine fibre analysis after optical microscopic and MSP 

analyses and concurs with the findings of Massonnett et al [36]. 

Vann et al [39] demonstrated the ability of Raman microspectroscopy to 

distinguish between rutile and anatase forms of titanium dioxide, an inorganic 

pigment, and to make quantitative measurements of titanium loading in fibres. 

Among the issues affecting the validity of the Raman measurements are the 

spatial heterogeneity of titanium dioxide in the fibre, the polarisation of the 

laser beam, and the polarising properties of the actual fibre. Levels of titanium 

dioxide in single, delustered polyamide fibres ranged from 0 to 7.1% titanium 

dioxide. Scrambling the polarisation of the laser minimised fibre polarisation 

and orientation effects. 

Zieba Palus et al [40] evaluated the application of the new combined micro- 

Raman and micro-XRF spectrometer to the analysis of multi-layer paint chips, 

contemporary inks, plastics, and fibres. The results obtained show that the 

apparatus has real advantages and could prove useful in the identification of 

examined materials after modifications such as the addition of an extra laser 

and reducing the spot size of the X-ray beam. 

As fibre and textile evidence is often encountered in forensic casework, it is 

important that the forensic fibre examiner is familiar with the different types of 

fibres - common and rare - on the market. This may prove challenging 

because the textile market is always changing, with new fibres, finishes, 

colours and shapes being produced. Akrap et al [41] examined the properties 

of aramid fibres, a high-performance fibre type not commonly encountered in 

forensic casework. 

Dines and McEvoy [42] evaluated method of fibre cross sectioning involved 

"threading" some of the fibres through a thin section of a shirt button and 

fixing them in place with superglue. The shirt button could then be affixed to a 

microscope slide and polished down to a very thin layer. This method was 

said to be fast, applicable to all fibre types and provide good fibre orientation, 

but was less suitable for short individual fibres. A second published method 

involved using a pipette tip as opposed to a button. This method was 

investigated and a variation based on combining the two methods proved 

effective and far easier than the current method, based on cellulose acetate 

slides. 

193

Morgan et al [43] studied the statistical validity of discrimination between 

various polymer classes and dyed textile fibres, visualisation of significant 

differences between groups of spectra discrimination, and tracking of spectral 

changes with environmental changes. Fibres and associated spectra 

contained in the database, combined with validated software, are useful for 

fibre comparisons in casework and for quality control and the training of 

analysts. This paper describes the application of linear discriminant analysis 

to a database containing more than 5,000 UV/visible absorbance and 

fluorescence spectra. 

Abendshien et al [44] Reported on the development of the FBI Forensic 

Automotive Carpet Fibre Identification Database (FACID). At present, FACID 

contains more than 700 automotive carpet samples from private vehicles and 

vehicle manufacturers. Questioned carpet-type fibres can be searched by 

microscopic characteristics, fibre type, and/or colour information in order to 

determine whether or not a particular type of vehicle was involved in the 

commission of a crime. FACID can also be searched by vehicle make, model, 

year, and/or colour in order to determine the carpet samples available for 

comparison. 

Ali et al [45] this study, a natural dye extracted from Eucalyptus camaldulensis 

was used to colour cotton fabric by direct dyeing under a range of different 

conditions. The aim of this work was to compare the fastness properties of 

dyeing by different dyeing techniques. This dye shows fairly good saturation 

on cotton and has medium-to-good fastness properties. 

Morgan et al [46] assessed whether forensic fibre discrimination could be 

improved by extracting the dye from the fibre prior to trace analysis by a highresolution 

separation technique. Methods developed for the extraction of 

unknown dyes from fibres, were based on three capillary electrophoresis (CE) 

procedures for dyes from six classes of textile dyes. CE with diode array 

detection is useful for the analysis of longer-length fibres, but a more sensitive 

and selective method, such as CE/mass spectrometry (MS) is required for the 

analysis of the small amount of dye (2-200 ng) associated with forensically 

relevant fibre samples. The analytical approach was destructive, but only an 

extremely small sample (approx. 2 mm of a single 15-um diameter fibre) was 

required. CE/MS analysis was found to be capable of separating extracted 

dye components and providing semi-quantitative estimates of dye amounts 

whilst providing qualitative information for the identification of dyes. 

Eng and Bhagwandin [47] investigated the effect of metamerism in the 

generation of different UV-visible spectra. Metameric samples of blue textile 

fibres, created using different colouring agents or different relative 

concentrations of the colouring agents, were examined in order to 

demonstrate their differentiation based on their spectra between 350 and 800 

nm using UV-visible microspectrophotometry. It was found that some of the 

metameric samples intersected at three or more loci, but some did not 

intersect at all. In the intersecting spectra, there was no correlation between 

either the dye chemistries of the relative component concentrations. 

194

Gallo and Almirall [48] applied microwave digestion inductively coupled 

plasma mass spectrometry (ICP-MS) and laser ablation inductively coupled 

plasma mass spectrometry (LA-ICP-MS) to the forensic elemental analysis of 

cotton fibres with the aim of improved discrimination. A cotton standard 

reference material was used to validate these methods, giving good accuracy 

with typically

8. The textile industry 

The present worldwide economic climate means that more than ever, the 

textile industry is in a state of flux, with availability and price or raw materials, 

increased costs of labour, transportation etc. influencing the industry. Despite 

this, all predictions are that the industry will expand, particularly in Asia and 

China. By far the most useful tool in monitoring developments in the textile 

industry (including the development and/ or emergence of new fibre types) is 

the internet. To this end, the URL’s listed in the reference section [51-60] are 

useful, but by no means exhaustive. 

9. The future 

From the case studies presented over the past three years, it is gratifying to 

see examples of where fibre evidence has been used extremely effectively in 

conjunction with other evidence types. Over the last few years fibre 

examinations have continued to demonstrate their value as a powerful 

intelligence tool in large complex enquiries, providing invaluable investigative 

steers. 

Practitioners, investigators and members of the judiciary need to consider a 

more holistic approach in the application of forensic science to the 

investigation of major crime. There has been a tendency for many to take a 

blinkered approach, rather than pausing to consider, “what question am I 

trying to answer?”, “What is the best way of doing this?” The future success of 

fibre examinations is not solely in developments in our understanding of the 

mechanisms of fibre transfer, or in the development of more discriminating 

means of analysis, but in an acknowledgement of how this evidence type can 

provide added value in combination with other evidence types. 

It has often been said that DNA evidence can provide the “who?” part of the 

equation, whilst fibres the “how?” and “when?” Whatever the accuracy of this 

statement, the truth of the matter is that proper case assessment and 

examination strategy planning, are the key to the future success and 

development of not only fibre evidence, but to forensic science as a whole. 

The EFG and SWGMAT will continue to be key in promoting and embedding 

this philosophy. 

10. Summary 

Despite the pressures on practitioners, the considerable amount of work 

relating to the forensic examination of fibres and textile materials over the last 

three years, continues not only to demonstrate a willingness to develop this 

evidence type, but also to address the cogent issues facing them. 

In the previous 2007 report [1], the author wrote “With the increased threat of 

terrorism, law enforcement agencies are continually looking to forensic 

science for the provision of intelligence in these (often) complex investigations 

196

- not only to identify the perpetrators of a specific attack, but also in preventing 

future incidents.” 

The ENFSI European Fibre Group has risen to the challenge and it is hoped 

that the “Fibre and Textile Evidence in Terrorist Cases - A guide for 

investigators.” document will become a useful aid in casework examination 

and strategy setting, not only in cases of terrorism, but in cases of major crime 

generally. 


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27. Palmer, R., Burch, H. J. The Population, Transfer and Persistence of 

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28. Chewning, D. D., Deaver, K. L., Christensen, A. M. The Persistence of 

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29. De Wael, K., Gason, F. Microfibre Transfer Experiments. Global Forensic 

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31. Jones J. Synthetic Flock Fibres – A Target fibre Study. Proceedings of 

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34. Biermann, T. W. Title Blocks of Colour IV: The Evidential Value of Blue 

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Blue Cotton. Sci. Just; 2009; V49 (1); March; P12-18. 

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from Eucalyptus. The Journal of the Textile Institute; 2007; V98 (6); 

P559-562. 

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Discrimination of Dyed Textile Fibers Using Capillary Electrophoresis / 

Mass Spectrometry. Proceedings Trace Evidence Symposium, 

Clearwater Beach Florida; 2007; August; P1-10 

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Their Forensic Significance. Journal of Forensic 2009; V54 (4); July; 

P841-845. 

199

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Evidence Using Solution ICP-MS and Laser Ablation ICP-MS (LA-ICP- 

MS). Forensic Science International; 2009; V190 (1-3); September; P52- 

57. 

49. Kirkbride, K. P.; Tridico, S. R. The Application of Laser Scanning Confocal 

Microscopy to the Examination of Hairs and Textile Fibres: An Initial 

Investigation. Forensic Science International; 2010; V195 (1-3); February; 

P28-35. 

50. Sano, T., Suzuki, S. Basic Forensic Identification of Artificial Leather for 

Hit-and-Run Cases. Forensic Science International; 2009; V192 (1-3); 

November; PE27-E32. 

51. http://emmergingtextiles.com 

52. http://fibre2fashion.com 

53. http://www.eurotexx.com/ 

54. http://www.fashion-links.de/ 

55. http://www.fashionseek.net/ 

56. http://www.ita.rwth-aachen.de/ 

57. http://www.techexchange.com/ 

58. http://www.texdata.com/ 

59. http://www.texi.org/ 

60. http://www.textile.fr/ 

61. http://www.textileweb.com/ 

62. http://www.textileworld.com 

63. http://www.textilexpert.com/ 

64. http://www.textilserver.de/ 

65. http://www.texweb.de/ 

66. http://www.ivc-ev.de/HTDOCS/index.html 

67. http://www.trevira.de/ 

Thanks go to Martin McEvoy for the assistance in carrying out the literature 

search. 

200

Forensic Geology 

Review: 2007 to 2009 

Ritsuko SUGITA, PhD 1) 2) , Hiroaki YOSHIDA, PhD 1) , 

and Hirofumi FUKUSHIMA, PhD 3) 

1) Trace Evidence Section, National Research Institute of Police Science, 

2) Identification Division, National Police Agency, 

3) National Research Institute of Police Science 

Corresponding author: Ritsuko Sugita 

National Research Institute of Police Science 

6-3-1 Kashiwanoha, Kashiwa-shi 

Chiba 277-0882, Japan 

201

TABLE OF CONTENTS 


2.  NETWORKING AND MEETINGS  203  

3.  BOOKS  204  

4.  ANALYSIS OF SOIL, SEDIMENT, AND ROCK EVIDENCE  204  

5.  BIOLOGICAL MATERIALS  207  

6.  TAPHONOMY AND SOIL  207  

7.  SEARCH OF CLANDESTINE GRAVES AND MISSING PERSON  208  

8.  EDUCATION  209  

9.  DATABASE AND EVALUATION  209  

10.  CASE REPORT  209  

11.  MISCELLANEOUS  210  

12.  ACKNOWLEDGMENTS  210  


202


This review article covers developments and topics in the field of forensic 

geology during the period from 2007 to 2009. The title of this review is “Forensic 

Geology,” but the review is not restricted in narrow definition of “geology”; it 

includes a wider meaning (1). Some practitioners have preferred to use the term 

“forensic geosciences” in recent years instead of “forensic geology.” This change 

in designation may indicate that more people of the field of geology and its 

related sciences are being noticed and getting involved in the science for 

criminal investigations. In this review, papers on environmental forensics have 

not been included. 

2. Networking and Meetings 

Networking was accelerated rapidly in the period between 2007 and 2009. The 

most significant movement was the establishment of GIN (Geoforensic 

International Network), which is a worldwide network of forensic geoscientists. 

Further, the international working group of forensic geology was agreed upon by 

IUGS-GEM (International Union of Geological Sciences – Geosciences for 

Environmental Management) in 2009 (2, 3). Dr. Donnelly had enthusiastically led 

forensic geoscientists around the world to build a network. Before the 

establishment of an international group, a key network of geoforensics was 

begun in the United Kingdom (GIMI: Geoforensics and Information Management 

for crime Investigation). There are also two ongoing projects in the UK. One is 

the SoilFit (Integration of Soil Fingerprinting Techniques for Forensic 

Applications) project (4), which is being carried out to investigate the potential of 

advanced analytical methods in providing soil forensic intelligence to police 

investigations. It is being carried out by a multi-disciplinary group of experts of 

the UK academic and law enforcement organizations. As part of this project, the 

group has examined various problems related to soil as both intelligence and 

evidence in a forensic context, such as establishment of a statistical framework, 

assessment of the variability of soil, and development of a geographic search 

tool. The other ongoing project is the SoilFUN (5) (Soil Forensic University 

Network), which was begun as a daughter project of the SoilFit project, based on 

the collaboration of universities and institutes in the UK. Morrison et al. (6) have 

reported the preliminary results of this project. A large number of undergraduate 

203

students enrolled in forensic science courses are learning how to apply their 

skills in a forensic context. 

The Second International Conference on Environmental and Criminal Forensics 

was held in November 2007 at Edinburgh (7). The conference included seven 

sessions, namely “environmental soil forensics,” “criminal soil forensics,” 

“geoforensics,” “geostatics, databases, and geographical information systems,” 

“biological and chemical analytical diagnostics,” “forensic taphonomy,” and 

“communications and advocacy” with a public lecture “the global way forward” by 

Dr. Robertson, and 31 presentations from the conference were published as a 

book titled “Criminal and Environmental Forensics” in 2009 (8). The third 

conference is planned in November 2010 in California, USA. FGG, the Forensic 

Geoscience Group of the Geological Society, held a meeting “Geoscientific 

Equipment and Techniques at Crime Scenes” in 2008 (9). In the annual 

meetings of the Geological Society of America (GSA), presentations on forensic 

geology were included every year (10). The topics cover a broad range of topics 

related to forensic geology, including education, analytical methods, and case 

reports. 

3. Books 

In addition to the book of Ritz et al. (8) introduced in the former section, at least 

three titles were published in the review period. Pye (11) published a book on 

this issue titled “Geological and Soil Evidence: Forensic Applications” in 2007. It 

describes the types of geological evidence, the techniques applied, and the 

evaluation of the obtained results for people not familiar to geology and for 

geologists who have little experience of forensic application. Ruffell and 

McKinley (12) released a book titled “Geoforensics” aiming “to show how various 

geoscience techniques are used in forensic investigations.” Tibbett and Carter 

(13) edited “Soil Analysis in Forensic Taphonomy: Chemical and Biological 

Effects of Buried Human Remains,” which is the first book on the chemistry and 

biology of soil related to the decomposition of human remains. 

4. Analysis of soil, sediment, and rock evidence 

Color examination is one of the fundamental methods of forensic soil 

examination. Sand and dune sediments from the beaches of Portugal were 

204

analyzed using spectrophotometer, and the results were compared (14, 15, 16). 

A unique attempt of color analysis was reported by Marumo (17) using 

computerized image analysis software. 

Morphological features of grains using an optical microscopes and/or a scanning 

electron microscope (SEM) were discussed by several researchers. The quartz 

grain surface was observed by SEM and examined to determine whether the 

surface texture changes at a car fire temperature by Morgan et al. (18). Millette 

et al. (19) examined coke and coal particles by observing them under both an 

optical microscope and an SEM in combination with an energy dispersive X-ray 

(EDX) analyzer. The analysis of the chemistry of light minerals in soil by an 

SEM-EDX system was reported by Aoki and Oikawa (20) to discriminate soil and 

an automated SEM-EDX to analyze grain chemistry; the shapes were described 

by Pirrie et al. (21, 22). Schwandt (23) presented a soil grain analysis using 

optical microscopes and digital x-ray maps, which could also provide information 

about the bulk chemistry of the sample by manipulation. The elemental data of 

feldspars sampled at the beaches of a prefecture obtained by an X-ray analyzer 

attached to an SEM was plotted on a mineralogical triangle diagram; the 

samples were successfully discriminated by the geological backland (24). The 

trace amount of soil and the smear of soil on cloth were examined by a 

variable-pressure SEM-EDX; and was considered useful for the screening of a 

forensic soil examination (25). 

Isphording(26) stressed the importance of heavy minerals even if the amount of 

these minerals in the soil was small because the types and their chemistry 

reflected their crystallizing conditions. Biotite and its weathered material are very 

common in immature surface soil in granitic rock regions. Their chemical 

composition reflected the composition of other heavy minerals stated by 

Isphording (26); this finding was considered to be useful information for forensic 

soil investigation (27, 28). Bowen (29) stated some principles of forensic soil 

comparisons and described the use of additional information such as 

morphology, chemistry, and isotopes of mineral grains to individualize soil 

samples. 

The use of cathodoluminescence was described to screen and discriminate 

mineral grains (30). Infrared (IR) spectrometry has long been used for providing 

205

the compositional and structural information of various types of substance, 

including minerals before the wide usage of X-ray diffractometry (XRD). It has 

been revisited and examined as a rapid identification method for mineral species 

in forensic evidence by Weiger et al. (31). Newly developed analytical 

techniques such as confocal Raman, laser induced breakdown spectrometry, 

and laser ablation inductively coupled mass spectrometry were introduced and 

compared to analytical methods currently applied to forensic geology by Walker 

(32). 

Pye and his co-researchers continuously improved an analytical procedure by 

inductively coupled plasma (ICP)–atomic emission spectrometry (AES) and 

mass spectrometry (MS) as already mentioned in the last review (1). Pye et al. 

(33) assessed compositional differences between different size fractions and 

different samples. The method was also applied to develop a searchable 

database by Pye and Blott (34). 

Isotope ratio mass spectrometry (IRMS) is a significantly developing field of 

forensic chemical analysis, and a network was recently established in the 

forensic community (35, 36). The application of IRMS analysis to determine the 

origin of carbonate rocks related to a crime was reported by Roelofse and 

Horstmann (37). 

The magnetic susceptibility of beach and dune sediments was measured for 

forensic application (15, 16). A unique application of magnetics used in the case 

of a car accident related to soil was reported (38) in which analytical results were 

obtained from the evidence and samples of two suspected origins, where the 

pedological features were very similar and were compared for discrimination. 

The work flow proposed by Graves (39) in 1979 was modified as a simple and 

easy-to-learn method (40). Iconology was introduced in addition to mineralogical, 

micropalaeontological, and petrographical studies to link a suspect to the crime 

in a simulation case work (41). A study utilizing X-ray fluorescence spectrometry 

(XRF), total organic carbon content, and ion chromatograph were performed to 

distinguish and estimate a region by Nagahama et al. (42) in a volcanic soil area. 

Melo et al. (43) applied a sequential extraction analysis for a forensic soil 

examination using various instrumental analyses such as ICP-AES, XRD, and 

visible ultraviolet spectrophotometry. Molina et al. (44) and Reyes et al. (45) 

206

examined the application of petrographical, mineralogical, and chemical 

analyses for characterizing soil for forensic purposes. 

The organic components of soil have not been considered to a great extent as 

compared to the physical and elemental composition of soil. Bommarito et al. 

(46) applied high-performance liquid chromatography (HPLC) and ion 

chromatography to discriminate soil evidence. 

5. Biological materials 

The importance of palynology was eagerly discussed in many articles during last 

period of review (1). Wiltshire (47) stated the requirement to a forensic 

palynologist. Nesterona et al. (48) reported cases using pollen in urban surface 

soil, which often changed by construction, for forensic investigation. The 

preliminary result of a unique attempt utilizing testate amoebae for the 

discrimination of soil was reported by Swindles and Ruffell (49). Testate 

amoebae was recovered from dried sediments on clothing 10 years after the 

case. Microfossil has been used frequently for environmental or age estimations 

in conventional geology. It is applied to discriminate concrete (50) in combination 

with a mineralogical examination. The plant wax in soil was examined by Mayes 

et al. (51) for the forensic discrimination using GCMS. 

Sensabaugh (52) overviewed the present state of microbial community profiling 

utilizing a DNA analysis in soil and discussed the challenges to overcome for 

forensic application. The spatial variation of bacterial DNA profiles was 

examined by Heath and Saunders (53) for a forensic soil comparison on soil 

samples obtained from three different ecosystems. As a result, small-scale 

variability can be a problem, but the precise location can be identified. Meyers 

and Foran (54) reported that bacterial DNA profiling may be useful although 

variables, especially the time difference, should be considered on the basis of 

the study on bacterial DNA in soil over a one-year period. Mcdonald et al. (55) 

examined the bacterial, archaeal, and fungal DNA in air-dried soil and found that 

fungal DNA was less altered than bacterial and archaeal DNA. 

6. Taphonomy and soil 

The number of papers on taphonomy with respect to soil environment has 

207

increased during this review period and includes the book edited by Tibbett and 

Carter (13). Carter et al. (56) reviewed the formation and ecosystem of gravesoil. 

Microbial activities and the chemistry of soil around a buried cadaver were 

studied (57, 58). Prangnell and McGowan (59) applied a soil temperature 

calculation method used in civil engineering to estimate the temperature of the 

burial site. The effects of the state of the buried tissue were studied. Stokes et al. 

(60), including changes in the chemistry and microbial activities in soil. 

7. Search of clandestine graves and missing person 

Searching clandestine graves by geophysical, geographical, and geochemical 

methods are now discussed eagerly. Ruffell and McKinley (61) spared a large 

volume to these issues in their book. 

Jervis et al. (62, 63) presented the result of an experimental survey on simulated 

clandestine graves considering ground water and soil data obtained by the 

electrical resistivity survey method. The result of the comparison of GPR and 

electron resistivity tomography (ERT) for the forensic search of clandestine 

graves was reported by Pringle et al. (64, 65). 

The effectiveness of the combination of geophysical and geochemical (including 

hydrochemistry) as well as the role of geoscientists in the search was described 

(66–69). Harrison and Donnelly (69) described the effectiveness, limitation, and 

devices of a geophysical survey, and the usefulness of hydrochemical and 

geochemical examinations. McKunley et al. (68) recommended the application 

of spatial methodology, including geophysical information system (GIS), GPR, 

hydrochemistry, and cadaver dogs to a forensic search. Parker et al. (70) 

reviewed the search of freshwater bodies using seismic waves, compressed 

high intensity radar (CHIRPS), side scan sonar, GPR, magnetometers, and other 

techniques for forensic purposes. An experiment to detect a clandestine grave 

using the difference in vegetation was also reported by Watson and Forbes (71). 

The examination of the use of nynhidrin to detect gravesoil was also presented 

(72, 73). 

The significance of this topic is that there are many case reports on this topic as 

compared to the other type of issues related to forensic geology. Ruffell et al. 

(74) reported a false-positive case of the application of ground penetrating radar 

208

and victim recover dogs to search for a suspected location as a “grave” of a 

missing person with a discussion. Ruffell et al. (75) described a search for 

historical mass graves using historical and anecdotal information, aerial 

photographs, GPR, and metal detectors. Congram (76) reported the prospection 

and excavation of a clandestine burial in Costa Rica utilizing a conventional 

search such as the slumping and different plant growth and by improvising a 

conventional archaeological excavation method. Billinger (77) introduced the 

case of the utilization of GPR for the location of a potential human burial under 

concrete. 

8. Education 

Forensic geology was introduced at different levels of education, including high 

school (78–81). In Colombia, “Ibero-American course on forensic geology” was 

conducted to assimilate geological methods for forensic investigations (82, 83). 

9. Database and evaluation 

Attempts to create a database for forensic purposes were reported. Pye and 

Blott (84) gathered existing data from their past examinations of elemental 

analyses in England and Wales to create a searchable database. The SoilFUN 

(4, 6) aims to produce analytical data on urban soils, which are relatively little 

known as compared to non-urban soils. Bergslein and Hovey (85) reported a 

progress on a project to establish a database on the soil in western New York 

using XRF and XRD. The difficulties and requirements were discussed by Aitken 

(86) to determine the likelihood ratio of the obtained data sets of the chemical 

analyses for a forensic soil examination. A geostatistical method was proposed 

to utilize spatial databases for solving an intelligence problem by Lark and 

Rawlins (87). 

10. Case report 

Cases related to geology and crimes are not only geological materials as trace 

evidence (88), but as objects of mining fraud cases (89). Criminal cases and 

fraud cases of mines, gems, and art were introduced to determine how geology 

can help in solving crimes by Murray (90). He also introduced a new and 

traditional microscopy technique applied to forensic geology. An experiment was 

209

eported to prove a continuous gas leakage from an underground gas pipe by 

the color of soil derived from the redox state of iron (91). Schneck (92) presented 

case studies in which a broad range of materials were involved. 

11. Miscellaneous 

Seismology is one of the very important fields of geosciences, but it is not 

considerably valued in the field of forensic science. It can provide the time, the 

scale of explosion, and additional information of an incident by using its network 

to help forensic investigation (93). Information on the persistence of grains was 

provided through case works (94) and experiments (95). The advantages and 

disadvantages of spatial sampling were discussed by McKinley and Ruffell (96). 

The trace evidence concentrator system was introduced for the separation and 

concentration of trace evidence from the soil by Smucker and Siegel (97). The 

system used a hydropneumatic elutriation method for a rapid examination. 

A review on the forensic sediment analysis was written by Morgan and Bull (98). 

The difficulties of communication are not the problem exclusive to forensic 

geology but of any occasion when specialists and non-specialists wish to share 

knowledge. Donnelly (99) reported on this issue with some case work and Rold 

(100) discussed the difference between geology and forensic geology. The 

status of forensic geology in Russian Federation was described in two reports 

(101, 102). Stam and Murray (103) compared the situation of the USA and the 

UK. Molina (104) reported the history of forensic geology in Colombia. 

There were also several papers written about the forensic geoscience for 

geologists, who were not involved in forensic science (105, 106). 

12. Acknowledgments 

We are grateful to Dr. Dawson and Dr. Donnelly for providing us with information 

about the networking. The members of GIN also offered us a considerable 

amount of useful information. Dr Suzuki assisted us throughout the reviewing 

process. 

210


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(GIN) 

http://www.geolsoc.org.uk/webdav/site/GSL/shared/pdfs/specialist%20and 

%20regional%20groups/Forensic/FGG%20Poster%202010.pdf. 

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211

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220

HUMAN IDENTIFICATION 

  221 

Fingermarks and Other Impressions Left by the 

Human Body 

Review :2007 – 2010 

Andy Becue, PhD, Nicole Egli, PhD, 

Christophe Champod, PhD, Pierre A. Margot, PhD 

Corresponding author: 

Prof. C. Champod 

Ecole de Sciences Criminelles / Institut de Police Scientifique 

Faculté de Droit et des Sciences Criminelles 

Quartier Sorge / Batochime 

Université de Lausanne 

CH-1015 Lausanne-Dorigny, Switzerland 

e-mail: Christophe.Champod@unil.ch 

http://www.unil.ch/esc 

222

ABSTRACT 

The purpose of this paper is to review the scientific literature from August 2007 

to July 2010. The review is focused on more than 420 published papers. The 

review will not cover information coming from international meetings available 

only in abstract form. 

Fingermarks constitute an important chapter with coverage of the identification 

process as well as detection techniques on various surfaces. We note that the 

research has been very dense both at exploring and understanding current 

detection methods as well as bringing groundbreaking techniques to increase 

the number of marks detected from various objects. 

The recent report from the US National Research Council (NRC) is a milestone 

that has promoted a critical discussion on the state of forensic science and its 

associated research. We can expect a surge of interest in research in relation to 

cognitive aspect of mark and print comparison, establishment of relevant 

forensic error rates and statistical modelling of the selectivity of marks’ 

attributes. 

Other biometric means of forensic identification such as footmarks or earmarks 

are also covered in the report. Compared to previous years, we noted a 

decrease in the number of submission in these areas. No doubt that the NRC 

report has set the seed for further investigation of these fields as well. 

We invite the reader to use two levels of reading for the report: a summary level 

and a more detailed level. In each subsequent section, if the number of papers 

published in a given area is large, the subsection will start with a quick summary 

(in italics) followed by a more detailed analysis of the research at hand. 

Copyright ©, registered ® and trademark logos were not included in the current 

text for better legibility. 

223


1  INTRODUCTION  225  

2  FINGERMARKS  228  

2.1  Friction ridge skin individualization process  228  

2.1.1  Automated fingerprint identification systems  228  

2.1.2  Papillary skin features  231  

2.1.3  The individualization process and the judicial system  238  

2.2  Fingermark composition, detection and visualization  243  

2.2.1  Composition, aging, and persistence of fingermarks  244  

2.2.2  Amino acid reagents  247  

2.2.3  Cyanoacrylate fuming  251  

2.2.4  Oil red O (ORO) & Physical developer (PD)  253  

2.2.5  Dry micron‐sized powders and powder suspensions  254  

2.2.6  Nanoparticles (nanopowders and nanocomposites)  257  

2.2.7  Antibody / Antigen recognition  259  

2.2.8  Detection of fingermarks on thermal paper  260  

2.2.9  Detection of fingermarks on metal and cartridge cases  261  

2.2.10  Detection of fingermarks on tapes/adhesives  263  

2.2.11  Detection of fingermarks on items collected from arson scenes  265  

2.2.12  Blood marks ‐ Fingermarks in blood and detection of bloodstains  268  

2.2.13  Fingermark detection and DNA or biological fluid analysis  269  

2.2.14  CBRNE‐related evidence (Chemical, Biological, Radiological, Nuclear, and Explosive)  270  

2.2.15  Detection and lifting of fingermarks on skin  274  

2.2.16  Miscellaneous detection techniques  275  

2.3  Forensic light sources, photography and digital/chemical imaging  279  

2.3.1  Alternative light sources (ALS)  279  

2.3.2  Photography  280  

2.3.3  Chemical imaging using Fourier transform infrared (FTIR) and Raman  281  

2.3.4  Miscellaneous imaging techniques  284  

3  MISCELLANEOUS  287  

3.1  Knuckles  288  

3.2  Barefoot impressions  288  

3.3  Ears and earmarks  289  

3.4  Lip marks  290  

4  CRIME SCENES AND CASE STUDIES  291  

5  REFERENCES  294  

224

1 Introduction 

This paper offers a review of the publication regarding marks left by the human body 

with an emphasis on fingermarks. It will cover the scientific literature from August 

2007 to July 2010. Because of the amount of material covered, this report does not 

intend to be a thorough critical review (unless it was felt required) but merely a point 

of focus and organisation of the available material. The material reviewed primarily 

originates from peer-reviewed journals and published books although non-reviewed 

electronic sources may at some points be used. Proceedings of colloquies will be 

used only when available in the form of full paper conference proceedings. More than 

420 references are covered here and, of course, we run the risk of having missed 

important material. In advance, we apologise to any author(s) or organisation(s) 

whose contribution(s) should have been mentioned. We also would like to thank 

individuals, peer groups and organisations that kept us informed of current 

developments and initiatives in the areas covered in this report. 

It is now customary to start our review by referring to the review by Brettell et al. in 

Analytical Chemistry (1) that contains also in some sections material that is in relation 

to the present topic. It can be complemented by (2), dealing with fingerprint detection 

techniques. 

A major scholarly publication in forensic science has been edited by Moenssens and 

Jamieson. This Wiley Encyclopedia of Forensic Science offers a range of papers in 

relation to fingerprints and human marks and the individualisation process: A 

discussion on individualisation is presented in (3). Aitken (4) presents different 

probabilistic tools used in forensic science (relative frequency, discriminating power, 

significance probabilities, likelihood ratio, and Bayesian networks). The author also 

discusses errors, e.g. the prosecutor’s fallacy, as well as a history of the different 

statistical tools used. Koehler (5) discusses how evidence is presented and 

understood in court. The author explains that a ‘match’ only indicates that no 

distinguishing characteristics were found between evidentiary marking and potential 

source, but does not indicate the evidentiary strength of this absence of distinction. 

Different ways of presenting statistical evidence are discussed as well as different 

misunderstandings of these statistics that can arise from the jury as well as the 

expert. More generally, Moenssens (6) discusses all demonstrative evidence (of 

which fingerprint evidence is a part if charting is used), and the necessity that the 

probative effect outweigh the prejudicial impact for the evidence to be admissible. 

One chapter is on observer effects in forensic science (7). Identification of human 

remains, including the use of fingerprints and techniques for obtaining fingerprints 

from decomposed, mummified or burnt bodies is the subject of a further chapter (8). 

Concerning fingerprints more generally, the anatomy of hands and feet as well as 

morphogenesis (9), comparison and identification of friction ridge skin (10), an 

overview of AFIS (Automated Fingerprint Identification Systems) (11) detection (12, 

13) and interpretational issues in the fingerprint domain (14) are included. Two 

chapters concern footprints: One is very general and includes shoe sole impressions 

(15), while the second concentrates on footprints of bare feet, feet in socks, and 

impressions inside of shoes (16). Ear marks and comparisons are treated in one 

225

chapter (17). Finally, a chapter on the photography of marks, impressions and 

documents is included (18) in this Encyclopedia. 

The field of biometrics is closely related to the topics under review. It is an impossible 

exercise to incorporate in this review all the scientific work carried out in this area. 

However a major Encyclopaedia has been published at Springer Verlag (19). An 

overview of the forensic applications (20) along with specific chapters on fingerprints 

(21-24), the Universal Latent Workstation (ULW) (25), cognitive processing in 

fingerprint recognition (26), earprints (27) and footprints (28) are covered in this 

Encyclopedia. 

A biometric handbook (29) offers also a good overview of the challenges in 

biometrics with one dedicated chapter on the relationships between forensic science 

and biometry (30). Another handbook but on forensic science (31) brings an overview 

chapter on fingerprints with specific emphasis on the situation in the UK (32). 

The forensic science landscape will certainly change after the publication of the US 

National Research Council of the National Academies (33). The report highlights 

some major causes of concern with regards to impression evidence (including friction 

ridge skin), to name a few: disparate training schemes, large subjectivity in the 

comparison protocol, recent cases of misattributions, evidence of cognitive bias and 

reporting practices that are tainted by a myth of infallibility. There is an urgent need to 

deal with some of the issues and ultimately increase transparency and accountability. 

The recent conference by the chairman of the committee, Judge H. T. Edwards, 

gives a very specific picture of the current situation with regards to the NRC report 

(34). An element of warning though: the report deals essentially with the state of 

affairs in the USA and does not necessarily reflect the situation in other countries. 

We can expect more contributions attempting to make explicit the complex set of 

psychological and cognitive processes involved in the identification process. The 

recent paper by Busey and Parada is a good example (35). Likewise, it is expected 

that the amount of statistical research devoted to these fields will still increase. The 

forthcoming paper by Neumann et al. (36) should provide the statistical foundation to 

the operational assessment tool developed by the Forensic Science Service in the 

UK. 

The NRC report received a very important press coverage including the most 

prestigious scientific journals such as Nature (37, 38) or Science (39). Professional 

organisations have reacted to the NRC report. We will retain here for example the 

statement by SWGFAST (Scientific Working Group on Friction Ridge Analysis, Study 

and Technology) (40). The IAI (International Association for Identification) also 

published a reaction to the NRC report. Here, the different recommendations in the 

report are reviewed from the association's point of view (41). The IAI furthermore 

sent a memo to its members, stating its support to many of the recommendations 

included in the report, cautioning against asserting “100% infallibility (zero error rate)” 

also advising its members not to state “their conclusions in absolute terms when 

dealing with population issues” (42). During the 2010 IAI conference in Spokane, the 

membership, following the recommandation of the Standardisation II committee, 

approved to rescind Resolution 1979-7 and Resolution 1980-5 (that prohibited 

examiners from providing less than certain testimony) and adopt a resolution 

acknowledging the recent progresses in fingerprint statistics: “The use of 

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mathematically based models to assess the associative value of the evidence may 

provide a scientifically sound basis for supporting the examiner’s opinion.” (43) That 

finally opens the possibility for fingerprint examiners to report strengths of evidence 

spanning over the whole spectrum of support in favour (or not) of an association that 

the observed features may bring. 

The NRC report stressed the lack of a structured research programme that underpins 

the identification process. We can only concur with this analysis; the resources 

attributed to research in the forensic identification disciplines outside DNA are indeed 

limited (this scarcity is somewhat due paradoxically to operational successes and 

proficiency, e.g. fingerprint evidence). Such research efforts (some are mentioned in 

this report) constitute the best response to future challenges. 

A framework decision of the council of the European Union requiring its members to 

ensure forensic laboratory accreditation according to EN ISO/IEC 17025 applies to 

laboratory activities resulting in DNA-profiles or dactyloscopic data has been 

published (44). This highlights the requirement to standardise forensic laboratory 

work. 

It is expected that the NRC report will also impact on how impression evidence, and 

fingerprint evidence in particular, is received in courts, especially in the US. A few 

fingerprint cases retain our attention here: In Langill, the court although retaining 

admissibility insisted on the need of contemporaneous case notes associated with 

the comparison process (45). In the case against Brian Rose, fingerprint evidence 

has been challenged, initially ruled inadmissible (46) and finally accepted by the court 

(47) even in the light of the NRC report. Both of these cases highlighted two 

important issues for the latent print community: documentation and error rate. The 

days of claims of 100% certainty in this area are counted. The case against J. Hull is 

a good example of this trend (48) (see the transcripts of the Frye-Mack hearing). In 

that case the judge accepted fingerprint evidence even though the experts did not 

ascertain that they could identify the defendant to the exclusion of all others. They 

merely put their evidence in the context of the limited relevant population. A 

commentary on this Frye-Mack hearing is presented by Bush (49). 

Finally the disputed identifications associated with the case of H.M Advocate v McKie 

are under review by a judicial inquiry (named the Scottish Fingerprint Inquiry). The 

associated report is due in the fall of 2010. All the documents used by the Inquiry can 

be found online: http://www.thefingerprintinquiryscotland.org.uk/. 

Another major contribution is the Fingerprint Sourcebook elaborated under funding 

by the National Institute of Justice (NIJ). It is published in stages at 

http://www.ojp.usdoj.gov/nij/pubs-sum/225320.htm. Currently five chapters have 

been put online. They deal with the history of fingerprints (50), the recording of prints 

(51), the fingerprint classification systems (52), the documentation of friction ridge 

impressions (53) and quality assurance (54). 

A special issue of Law, Probability & Risk presents a good overview of various 

arguments currently present in the fingerprint domain (55-60). Its content will be 

covered in detail in this report. Fingerprint comparison and DNA analysis are aptly 

juxtaposed in a recent book by Lynch et al. (61). 

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A few general works on human identification have been published during the review 

period. A history of personal identification, focusing on anthropometry, fingerprinting 

and DNA from the angle of racism and the different concepts and uses of race in 

identification is presented (62). Another historical review, but centred on the increase 

in the use of biometrics, is proposed by Maguire (63). The forensic use of marks left 

by papillary surfaces as a means of human identification is often traced back to 

Henry Faulds. Recent historical research has shown that a French chemist, Paul- 

Jean Coulier, suggested its use as early as 1863 (64). He made use of iodine fuming 

to detect surface alterations on questioned documents. Concurrently he observed the 

detection of latent fingermarks. At that point he proposed them as a means to identify 

the forger of the document. 

2 Fingermarks 

2.1 Friction ridge skin individualization process 

This section starts with publications on AFIS (Automated Fingerprint Identification 

Systems). Here, in particular, collaborations between countries (the Prüm treaty 

implementation), the reduction of the number of candidates to match finely through 

database indexing or filtering, and tests on different automated systems are 

presented, as well as studies on the interaction between the human fingerprint 

examiner and the automated system. 

The second part concerns fingerprint characteristics; studies concerning level I, II 

and III details (including frequencies of various characteristics, sex differences, 

studies aiming at estimating probabilities of random correspondence, improvements 

in matching rates) are presented here, as well as works on morphogenesis, 

influences on fingerprints, and the biological use of ridges. As far as possible, studies 

are presented by decreasing generality. Finally, the section on the individualization 

process and the judicial system presents first general works on considerations 

concerning individualisation, then errors, studies on bias, and discussions on the 

validity of fingerprint comparison and debates as well as studies concerning ACE-V. 

2.1.1 Automated fingerprint identification systems 

A general paper on fingerprint matching (65) and one on palmprint 

matching (66) have been published. 

Precisions concerning the implementation of the Prüm treaty are available 

(67-70). Two articles pertaining to the US-VISIT programme are integrated 

into this review (71, 72). The connectivity of systems is treated in (73), (74) 

and (75). Two trials involving several system providers are published, 

where the second investigates the usefulness of integrating extended 

fingerprint features (76, 77). Database filtering, indexing and clustering 

has been treated in the following articles: (78), (79), (80), (81), (82) and 

(83). The interaction between humans and automated systems is finally 

discussed and researched (84-86), including the use of expert 

assessments for improving the system (87). Image compression is the 

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subject of several articles (88-91) and (92) analyses a risk in image 

enhancement. 

The incorporation of the Prüm treaty is defined as follows: a decentralised system will 

be used, where each nation has its database, and the contact points of other nations 

will have access to reference data (67). A detailed description of how this data is to 

be exchanged (format, communication channels) is provided (68). Estimated daily 

maximum throughputs for the different countries (for different search modalities) are 

available (69). Good practice for such searches is proposed (70); e.g. to start with the 

most relevant searches, and to grade the importance between crime types. 

An overview of past work on fingerprint quality in US-VISIT is presented (72). A 

performance improvement by using two fingers and two stages (rather than 

increasing the number of fingers used to ten) for US-VISIT has been proposed. This 

two-stage process is compared to a two-stage process including two fingers and the 

face (71). The two-stage process for fingerprints consists first in a minutiae-based 

matcher and second on a texture-based matcher. The detection probability is 

increased with respect to minutiae based matching both by using minutia matching 

and face and minutiae and texture matching, but more so for the second option. 

The challenge of the increasing use of data interchange has been thematised. 

Discussions of both data interchange between countries (73) and between states, 

cities and federal government (74) have been published. This second reference also 

includes a historical overview and discusses connectivity between systems, which is 

opposed to interoperability (74). The interoperability between systems from different 

vendors and related challenges are described and investigated (75); in particular, the 

reduction of information in standard-compliant minutiae templates (as opposed to the 

proprietary format) significantly increased error rates in the different systems used, in 

a test carried out on a total of 4041 index fingers. 

A comparison between different systems has been carried out (76). The different 

technology providers who participated in the trial were Motorola, Inc., Sonda 

Technologies, Ltd., NEC Corporation, Peoplespot, Inc., SPEX Forensics, Inc., 

Cogent, Inc., L1 Identity Solutions and BioMG, Ltd. Each technology provider 

submitted a software development kit (SDK), and the submission of research 

algorithms was encouraged for the study. The test set consisted of 835 marks and 

their ten-print mates, showing sufficient information to result in an identification 

conclusion. Two background databases were also used, one containing 5000 records 

and one containing 10000 records (50000 and 100000 fingerprints). The rank 1 and 

rank 10 identification rates are reported, as well as other measures. Particularly 

interesting is the fact that both the original images at 1000 dpi and the same images 

downsampled to 500 dpi were used. Most software development kits showed an 

increase in performance when 1000 dpi images were used (as opposed to 500 dpi); 

this improvement was not huge, however. Also, while some hits were gained with the 

increase in resolution, some others were lost. A related short communication has 

also been published (93). 

Database filtering, indexing and clustering has been treated in the following articles: 

(78) and (79) use singular points for database filtering; indexing is proposed by (80), 

(81) and (82), and finally database clustering is the subject of (83). The specialised 

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literature is so abundant in this area that we cannot hope for exhaustiveness here, 

but it was felt important to provide some entries dealing with this subject, especially 

when the research work proposed has the potential for a change in practice in the 

short term. 

Matching performance is examined when plain as well as rolled impressions are 

used, and different fusion techniques are investigated, at feature, score, and rank 

level (79). Boosted max fusion at score level performed best; here, a rank-1 

identification rate of 83% was obtained, which compares favourably to the rank-1 

identification rates of 57.8% for plain and 70.4% for rolled impressions. 

The NIST engaged in collaboration a range of AFIS providers and parties in the 

definition of a new exchange format adding an extended set of fingerprint features: 

dots, incipient ridges, ridge edge protrusions and pores. All relevant documents are 

available on http://fingerprint.nist.gov/standard/cdeffs/. The new standard ANSI/NIST- 

ITL 1-2010 (now in draft version 0.4) will replace the ANSI/NIST-ITL 1-2007 and 

ANSI/NIST-ITL 2-2008 standards that address the interchange of fingerprint, facial, 

and SMT data). 

The use of this extended feature set has been the subject of a preliminary report 

(77). The participating organizations of this trial are SAGEM, NEC, Cogent, Sonda 

and Warwick. The number of marks used was 1114, and here, the mates had not 

been found using an AFIS search. The different features of the marks used were: 

image only; annotated region of interest (ROI); ROI, pattern class and quality map; 

minutiae with ridge counts; extended features; extended features with skeleton; and 

minutiae with ridge counts but without the image. Human markup was used for the 

marks, while automated feature extraction was used for the 100000 (ten-print) 

records used as a background database. When extended features are added, 

performance is improved only for a subset of participants. 

Kwan and co-workers propose to incorporate expert feedback into AFIS (87). 

Relevance feedback is proposed as an add-on module to automated systems. The 

examiners decisions (‘positive’, when the impressions are judged similar, or 

‘negative’ otherwise) are used to transform the input space. When the examiner is 

faced with a list of possible matches, the positive match declared will be used to 

decrease the distance (or increase the score) by a certain amount, while a negative 

declared will do the opposite. A new comparison will then be mapped onto a 

semantic space created on the basis of the transformed distance (or score) matrix; in 

this way, the short-listed results will be increasingly similar with respect to the 

examiners assessment. 

The use of large, computerized databases may lead to the discovery of impressions 

that come from different fingers that show more similarity than what was expected 

before the use of AFIS searches. This is the subject of an article by Dror and 

Mnookin (84). In particular, the authors consider that the way fingerprint conclusions 

are arrived at (in particular the sufficiency requirements for an individualisation) 

should change with the use of AFIS. They also discuss the possible introduction of 

bias through the candidate list and the associated scores. The interaction between 

the examiner and the automated system is also the subject of an article by Wertheim 

(86). The different necessary steps for a successful search in an AFIS were followed 

on a set of 1368 images of marks. These steps were formatting (proper orientation 

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and scaling of marks), encoding (annotating minutiae and singular points in the 

universal latent workstation (ULW)), and comparison (submitting encoded searches 

to various databases and examining the candidates proposed, identifying one of the 

candidates if warranted). Failures occurred at all levels; however in the comparison, 

only false negatives were observed. At least some of these false negatives were 

caught by a blind verifier; the rate of individual examiner false negative errors was 

14%. Furthermore, almost half (7 out of 15) of the examiners missed one or more 

individualisations in this way, which shows that false negatives may happen more 

frequently than previously thought. 

Davis and Hufnagel have investigated the personal perception of NAFIS, before and 

after its implementation. The study was carried out on a small sample of fingerprint 

experts and younger fingerprint technicians (not yet having expert status). The 

organisational, role, task and process level changes, as expected or perceived by the 

fingerprint examiners, are exposed (85). 

Wireless transmission of fingerprints for operational purposes requires the images to 

be compressed; experiments have been carried out to determine the optimum form 

of compression, allowing realistic transmission times without compromising AFIS 

searches (or storing of the fingerprints in AFIS) (88). Image compression is also the 

subject of two further articles, where a proposed algorithm is compared to the 

compression algorithm used by the FBI (89, 90). Lossy compression standards, as 

well as evidence acquired and used in a digital format in general and image 

processing in this context, are criticised by Cherry and Imwinkelried (91). The 

probability of creating a false minutia (leading possibly to a false positive) is 

computed by estimating the probability of creating a square of pixels of the width of 

ridge, with grey values that are within 10 of the grey value of a ridge (for an 8-bit 

scale; for a 16-bit scale a difference of 2560 is used) by using an innocent image 

processing method. This probability is vanishingly small (92). 

2.1.2 Papillary skin features 

Both genetic (94) and environmental (95) factors influencing the 

development of friction ridge skin have been investigated, and a review 

presented (96). How ridges are linked to mechanoreceptor response is 

treated in (97),(98), and (99). Links between general characteristics of 

fingerprints and gender identity disorders (100), schizophrenia (101) as 

well as intelligence (102) have been investigated. 

Finger number and general pattern allow the distinction between two 

populations in the same country (103). Gender differences in general 

characteristics of fingerprints, mostly ridge densities, are reported for a 

large number of different populations (104-111); these data show that 

while such differences are rather easily interpretable for a single 

population, the values diverge between different populations and 

therefore, gender discrimination is less easily applicable when it is 

unknown which population the donor comes from. The selectivity of handshape 

is shown in a biometric context (112). Creases are the subject of 

two publications showing their usefulness: While (113) demonstrates 

selectivity, Zhou et al. (114) furthermore investigate the permanence of 

creases qualitatively. Two studies use AFIS in the context of fingerprint 

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individualization (115, 116). Several generative models for fingerprint 

individuality have been proposed (117-124). The addition of ridge points to 

minutiae has been evaluated with respect to matching performance (125, 

126), and minutiae frequencies have been presented (127). An operational 

trial on an evaluative model has been carried out (128). Finally, twin 

studies show that twins’ fingerprints are closer than randomly chosen 

ones, but distinguishable (129, 130). 

With respect to level III features, several studies concern their integration 

into the matching process (77, 131-137), one study assesses fingerprint 

examiners’ opinions on such features, and one study concerns the 

reproducibility of pore areas (138). Distortion has been qualitatively 

evaluated (139) and quantitatively modelled (140), and a different type of 

artefacts appearing in marks is discussed (141). 

Morphogenesis 

In order to gain a better understanding of the genetic modes of dermatoglyph 

inheritance, the dermatoglyphic traits after population admixture have been studied. 

The populations studied are Liujia Han, Kam and offspring of intermarried Kam and 

Liujia Han couples (China). Such marriages were not culturally permitted until 

recently. In particular it is observed that some traits either increase with respect to 

either of the parent populations (finger ridge counts and the whorl pattern) or 

decrease with respect to these parent populations (the frequency of arch patterns). 

These observations allow to suggest genetic pathways of the transmittal of these 

dermatoglyphic traits (94). A review of studies concerning the ontogeny of friction 

ridges is presented, concluding that these studies explain how the embryonic 

stresses and tensions result in the uniqueness of friction ridges (96). A study carried 

out using a sample of Dutch adults, identified through records of urban births during 

1943-1947, showed that environmental factors (war, famine) influence the difference 

in ridge count between the first and the fifth digit (95). 

Biological studies 

The influence of papillary ridges on Slowly Adapting type I (SA-I) mechanoreceptors 

in encoding edge discontinuities has been investigated; three necessary criteria of 

the lever-arm functionality are subjected to finite-element analysis. This results in 

discounting the lever-arm model, and a revised functionality is proposed (97). The 

function of ridges with respect to mechanoreceptors (spectral selection and 

amplification) is investigated using a biomimetic tactile sensor by Scheibert et al. 

(98). Here, the mechanoreceptors investigated are Pacinian corpuscules. In reaction 

to this article, Dahiya and Gori, while acknowledging the added insight gained by this 

study, highlight the fact that skin structure is more complex than the model used by 

Scheibert et al., and that therefore their study cannot conclusively answer all 

questions relating to the effect of skin structure on receptor responses (99). 

Links between fingerprints and other characteristics 

Several studies aiming at establishing links between psychological and physical 

attributes (e.g., fingerprints) have been published (100-102). These links are at most 

statistical and must be interpreted prudently. 

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Studies on general characteristics 

A study on two populations (two cities in the same country) using a classification and 

regression tree algorithm is presented (103); here, the goal is to use only finger 

number and general pattern for the prediction of a population that an individual 

belongs to. The classification is expected to be correct, when using all ten fingers, 

90.8% of the time if all ten general patterns of the individual are left loops or whorls 

with only the right ridge count, and 98.4% if all general patterns of the individual are 

whorls with two ridge counts. 

Frequencies of various dermatoglyphic measures for males and females of Kavalan 

(one of the Taiwan aboriginal populations) are reported by Chen et al. (104). These 

include general patterns, identical patterns on the corresponding fingers of the right 

and left hand, corresponding patterns on all fingers of one hand and on both hands, 

ridge counts, atd angle, tPD, palmar interdigital, thenar and hypothenar patterns, 

different combinations of palmar interdigital, thenar and hypothenar patterns on left 

and right corresponding hands, angles and simian line on palms. 

Ridge density in a sample of Spanish Caucasians is also reported (105). Ridge 

densities of 16/25 mm 2 or less in the radial area considered are more likely to be of 

male origin when using equal priors, while ridge densities of 17/25mm 2 or more are 

more likely to be of female origin; for the ulnar area these numbers are of 14/25mm 2 

or less for males and 17/25mm 2 or more for females. Sex-differences in an Indian 

population are reported by Nayak et al. (106) ; here, a mean ridge count of 12/25mm 2 

or less is more likely to be of male origin while a greater ridge count is more likely to 

be of female origin. Ridge densities for a South-Indian population are reported by 

Gungadin (107), where 13/25mm 2 or less are more likely if the impression is of male 

origin, while 14/25mm 2 or more are more likely if the impression is of female origin. 

Nayak et al. also report on ridge densities for Chinese and Malaysian populations 

(108, 109) ; here, in the Chinese population, a ridge density of less than 12/25mm 2 is 

more likely to be of male origin while ridge densities above 13/25mm 2 are more likely 

to be of female origin. In Malaysian subjects, it is a ridge density of 11/25mm 2 or less 

that is more likely to be of male origin, and a fingerprint with a ridge density of more 

than 13/25mm 2 is more likely to be of female origin. The relationship between ridge 

density and stature has however not been investigated. Furthermore, fingerprint 

classification and gender distribution was studied in a South Indian population (110), 

and no difference by sex between general patterns was highlighted in this study. The 

reason for the observed difference between sexes as to ridge density is the object of 

a letter to the editor (111). Data in relation to the general pattern present in the feet 

have also been published (142). Duta (112) shows the selectivity of the hand-shape 

as a biometric. 

Creases 

An automated extraction and comparison procedure for palmar flexion creases, 

carried out on a sample of 100 palms with 10 images of each one, achieves a 

genuine acceptance rate of 100%, for a false acceptance rate of 0.0045% (113), 

thereby illustrating the selectivity of this characteristic. Crease detection has been 

investigated by Zhou and co-workers (114), with particular application to the 

fingerprints of elderly people. The stability of creases is investigated, and matching 

based on creases and minutiae carried out. Also, the creases are used to remove 

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spurious minutiae around the crease, within a distance of 5 pixels. The combined use 

of minutiae and creases diminishes error rates in matching with respect to matching 

based exclusively on minutiae, but only for the database of elderly people. On a 

general population database this improvement is only very small. 

Level II detail 

The best way of analysing data in the form of match scores from a fingerprint 

matcher has been the object of two articles (143, 144). ROC (receiver operating 

characteristic) curves are compared to likelihood ratios (LR). Here, a LR above one 

yields the classification result that two impressions belong to the same person and a 

LR below one that they belong to different persons. For obtaining LRs, the within-and 

between-finger variabilities are modelled using gamma- and normal distributions. The 

likelihood ratio method outperforms ROC curves, but the performance of the two 

methods is similar when large numbers of minutiae are available (143). However, an 

analysis of four different matching systems shows that there is no definitive 

underlying distribution function for match and non-match scores. Nonparametric 

analyses on the discrete distribution functions of the four sets of scores (issued from 

the four different systems) are presented, and take the form of ROC curves (144). 

Note that while Srihari and Srinivasan (143) privilege LRs over ROC curves for 

performance reasons, Wu and Wilson (144) use ROC curves in the nonparametric 

approach. This second approach is, according to these authors, the proper way of 

analysing such data, due to differences in the distributions obtained when different 

matchers are used on one hand and the fact that the data is discrete (or can be 

transformed to discrete data) on the other hand. 

A study on the variability of minutiae in a Spanish population is presented by 

Gutierrez and al. (127). The authors studied in particular the total count of the 

minutiae, the count of each different type (where combined minutia types were used), 

both in the whole fingerprint and in the centre (defined as a circle with a radius of 18 

ridges) and periphery separately. The frequencies of the different minutia types are 

given. Also, the number of minutiae on the fingerprint is compared between male and 

female subjects. Differences are observed for the whole fingerprint, when the general 

pattern is whorl or loop, and differences are significant for all general patterns in the 

peripheral region, males having more minutiae than females. In the central area, 

none of the differences observed between male and female subjects are significant. 

Zhu and co-workers (123) use minutiae location and orientation in a generative 

model. This work and related ones has been reviewed also by Dass et al. (23). Here, 

a joint distribution is proposed, where minutiae locations are modelled by a bivariate 

normal mixture, and orientations by a Von-Mises distribution. A different approach, 

also using the model of Zhu et al. (123) as a basis, integrates image quality by 

adding a model for errors in minutiae detection and localisation (124). An internal 

report gives an overview of models for fingerprint individuality (145), proposing a 

classification of models into grid, fixed probability, ridge-based, relative 

measurement, and generative models. Generative models for birthdays, height and 

fingerprints are described (117). The fingerprint generative model includes minutiae 

locations and directions, as well as ridge lengths and the location and direction of 

ridge points, where one ridge point is selected in medium ridges and two ridge points 

for long ridges (none for short ridges). The parameters of the model are then 

estimated on the basis of data (100 fingerprints and 8 impressions from each 

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fingerprint, FVC2002 DB1), and the model is used to compute a probability of 

random correspondence given the number of minutiae and ridge points in the 

reference and query images, as well as the number or correspondences between the 

two. This model is further described by Su and Srihari (118) : a mixture of bivariate 

Gaussian distributions is used for minutiae location, von Mises distributions are used 

for minutiae orientation and the ridge lengths are considered to be distributed 

uniformly. Ridge points are described by the distance from the minutia, the direction 

between the minutia and the ridge point as well as the orientation of the ridge point. 

The distance of ridge points to minutiae is modelled using a one-dimensional 

Gaussian distribution. The model, with a computation of specific random 

correspondence probabilities, is the subject of (119). 

Chen and Moon carry out an investigation of discriminative power using minutiae 

data (location, direction). Minutiae location is modelled using a uniform (spatial) 

distribution, and a Von Mises distribution is used for the differences between 

orientations of two compared minutiae. First, the model for locations is compared to 

observed values (obtained on different available fingerprint databases), and second 

the model for both locations and directions is compared to known nonmatchcomparisons. 

The ‘score’ is constituted of matching pairs of minutiae between the 

(nonmatching) impressions. Good correspondence between the model and 

observations are shown (121). However, in a following paper, the authors show that 

the location of minutiae is not distributed according to complete spatial randomness, 

invalidating this first model (122), and the authors propose to model the minutiae 

locations using a quantitative stochastic model. The results obtained are again 

compared to observed data as well as to the first model, and an improvement (better 

correspondence to empirical data) is observed. 

The neighbourhood of minutiae is analysed by Hsu and Martin. The locations of 

minutiae are shown to be non-random (a preference of four inter-ridge distances is 

observed). The relative minutiae orientations and positions are distributed nonisotropically. 

Probabilities for various numbers of minutiae and numbers of nearest 

neighbours are given. For example, the probability for 12 minutiae is inferior to 10 -30 . 

The authors also show that manually annotated minutiae yield results that differ from 

those obtained on automatically extracted minutiae (120). 

Ridge points are described in (125), and their use for improving fingerprint matching 

is assessed using a modified version of the Bozorth and k-minutiae matchers. 

Minutiae types and orientations, as well as ridge types (defined using the ridge points 

described in (125)) are used in order to compute a PRC (probability of random 

correspondence) for fingerprints by Fang and co-workers (126). 

Two studies using AFIS (Automated Fingerprint Identification System) in the context 

of fingerprint identification have been carried out. One makes use of AFIS scores to 

determine the validity of fingerprint identification (115). Comparing mated inked 

prints, and using a logistic regression classifier, a threshold is used to declare a 

match. When this threshold is set to a probability of 0.5, the classification error is of 

2.4 ± 0.1 %. When marks acquired by research assistants were used in a second 

experiment, this classification error was of 4.5 ± 0.4%. In the third experiment, where 

marks from a NIST database were used (NIST Special Database 27), a classification 

error of 5.4 ± 1.0% was obtained (115). 

235

The second study aims at obtaining likelihood ratios from AFIS scores in the context 

of 1:1, mark-to-print comparisons. Here, a within- and between-finger variability are 

modelled, based on the characteristics of the mark and the suspects’ fingerprint 

examined. All computations are conditioned by the number of minutiae that are in 

agreement between the mark and the print (116). Two donors were used for the 

establishment and testing of this model. The rates of misleading evidence in favour of 

the prosecution (LRs above one when the impressions compared come from different 

fingers) for donor 1 (donor 2) are of 0.3% (5.2%), 0.3 (3.2%), 0.3% (0.8%), 0.0% 

(0.2%), and 0.0% (0.3%) for 6, 7, 8, 9 and 10 minutiae respectively, while for these 

same numbers of minutiae, the rates of misleading evidence in favour of the defence 

are of 0.7% (2.1%), 0.3% (0.3%), 0.3% (0.6%), 1.2% (0.00%), and 0.1% (0.00%). 

The background database used to extract the between finger variability consists only 

of images from the same finger number and general pattern as that of the finger that 

left the mark. An operational trial using a model to evaluate cases has been 

presented (128). 

Twin studies 

A trial has been carried out on a new set of 227 pairs of identical (188 pairs) and 

fraternal (39 pairs) twins’ fingerprints. Prints were acquired using a Smiths-Heimann 

scanner and processed using Cogent software. First, the probability that level 1 on 

the same finger of a pair of identical twins matches was computed and is of 66.71% 

(46.41% for fraternal twins). The MINDTCT algorithm was used for the extraction of 

minutiae and the Bozorth matcher for the computation of match scores. Distributions 

of scores are compared for twin-to-twin, twin-to-nontwin and genuine matches (where 

two impressions from the same finger are compared). These distributions all differ 

significantly. Error rates are also compared between non-twins, fraternal and identical 

twins ; these are 3.33%, 4.88% and 5.09%, respectively (129). Another study of twins 

(298 twin-pairs, 42 of which are fraternal twins) also shows that twins’ fingerprints are 

more likely to show the same general pattern for a given finger (56.92% for identical 

twins, 39.44% for fraternal twins and 31.76% for non-twins. Using the MINDTCT and 

Bozorth algorithms, the equal error rate for twins obtained is 6.17%, and 2.91% for 

non-twins. Testing of the distributions of match scores shows that the distribution for 

twins is different than that for comparisons between impressions from the same 

finger. The distribution for fraternal twins does not differ from that of identical twins, 

and the statistical test used does not formally show a difference between the 

distribution for twins and non-twins; it is concluded however that the similarity of 

fingerprints of twins is different from the similarity between arbitrary fingers (130). 

Level III detail 

A survey among practitioners has been carried out in order to clarify the definition 

and potential value of level III features. Disagreements in definition could largely be 

attributed to the wording or misunderstandings in the questions, but large differences 

were observed among examiners concerning the expected reproducibility of level III 

features, as well as their discriminative value (146). 

A large trial on level III features has been carried out, using pores, ridge contours and 

edge features. Matching capabilities are improved slightly by integrating level III 

features in addition to level II features (136), using the Bozorth matcher. A short 

overview of this trial has also been published (137). 

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A doctoral thesis (131) investigates additional features for use in fingerprint 

comparison, including ridge skeletons, pores, dots and incipient ridges. The matching 

performance of algorithms is improved by the addition of these features. When 

matching is performed in mark to inked print tasks (at 500 dpi) the improvement is 

largest for the worst images, and for ridge skeletons. These effects are attributed to 

the resolution of the images, which is small for the extraction of pores, in particular. 

When the comparisons are carried out between livescans, the improvement is 

greatest for the smallest areas used. In a second part, a fingerprint individuality 

model is proposed, incorporating minutiae locations and directions, ridge period and 

curvature as well as pore spacing. This model is an extension of one of the models 

presented above (123). The current model is fitted to five major fingerprint classes 

(whorl, left and right loop, arch and tented arch). Minutiae are clustered according to 

position and orientation, then position is modelled using a bivariate Gaussian 

distribution for each cluster, and the orientation by a Von-Mises distribution. The 

ridge period is modelled using a Gaussian mixture, and ridge curvature is modelled 

using a Poisson mixture. Finally, pore spacings along the ridge are modelled using a 

Gaussian distribution. Estimation of the model parameters follows, and the model is 

validated against empirical data. In the last part of the thesis, a touchless 3D 

fingerprint acquisition device is presented. In particular, the interoperability between 

these 3D images with legacy rolled prints is achieved through an algorithm 

constructing rolled-equivalent images from the 3D representations acquired by the 

sensor, and another to enhance the acquired images. The work carried out during 

this thesis is also the object of several articles (132-134). 

The usefulness of integrating level III features into the matching process is also 

shown by Vatsa et al. (147), Zhao et al. (148, 149) and Jain et al. (135). In this last 

reference, ridge edges as well as pores are used, in a hierarchical approach (level II 

is matched, and then level III features are used). Matching results are improved using 

this approach, also for images classified as being of low quality. But Indovina and 

Hicklin (cited above, under Automated Systems) only find an improvement for some 

of the systems tested, therefore highlighting that level III is not always useful in the 

matching stage (77). 

Pore area reproducibility has been measured on marks developed using ninhydrin 

and cyanoacrylate. Ten marks developed using ninhydrin and 50 marks developed 

using cyanoacrylate were used. The results show that pore area is not reproducible 

(138), indeed, the % coefficient of variance is between 38.5% of the mean and 

81.9%. 

Distortion 

The deformation of the skin under distortion conditions is observed experimentally by 

Maceo (139). First, the anatomy of the hand is described in a detailed fashion, 

describing the elements that are relevant to distortion. Then, the effects of vertical 

and horizontal sheering stresses and torque are examined, first on stills from videos 

and second on developed impressions. The two fingers used present different 

general patterns (loop and whorl) and differences are analysed qualitatively. This 

article is richly illustrated with commented images. A model for skin distortion in a 

biometric setting is proposed by Maltoni and Cappelli (140) (as well as its use for 

generating fingerprints and a discussion of the use of distortion to detect fake 

237

fingerprints). Butterflies (merging, overlapping or misalignment of two sections of 

ridge skin) are discussed by Pierce and Turnidge (141), who present examples of 

such merged impressions, and discuss how to detect their presence and interpret 

artefacts due to this kind of distorsion. 

2.1.3 The individualization process and the judicial system 

General considerations of the individualisation process are presented in 

(150) and (151). Errors are mentioned in several contributions (152-155); 

the definition of erroneous exclusions is discussed in (156). The influence 

of different evidence types (including fingerprint evidence) on verdicts is 

studied in (157). A change in the interpretation of fingerprint 

individualisation conclusion is apparent in (158, 159) and discussed in 

(160); these conclusions are, according to these publications, considered 

opinions (rather than facts). Bias is the subject of several publications 

(161-171); here (162), (163) and (165) present study results, and those of 

(162) and (163) show in part similar effects, while the third study has been 

criticised (166, 167). The validity of fingerprint individualization (i.e., the 

correctness of results) as well as the definition of ACE-V and whether or 

not it is scientific are all questioned (56, 58-60, 172-178) (criticised in 

(179))(180). Kaye makes interesting points on uniqueness and 

individualization (181, 182). Two articles spell out the ACE-V process 

(183, 184), while one study tests the validity of conclusions resulting from 

the process (185), and one other tests the validity of conclusions on 

laypeople (186). Finally, Cooney (187) inventories latent print training. 

Koehler (188) shows shortcomings of current proficiency tests for 

computing error rates and proposes solutions. Koppl (189) analyses the 

relative cost of errors and systematic blind verification for felony cases 

going to trial, and advocates blind verification. 

A decision theoretical approach is proposed for the identification sciences in general. 

The problem of how to come to a definitive decision (inclusion, exclusion, 

inconclusive) using probabilities and utilities is described and elegantly solved (150). 

The increase in the number of fingerprints (in the sense of the number of people on 

earth increasing, and not solely of the increase in database sizes) and the possible 

problems arising from this increase is the object of an article by Cherry and 

Imwinkelried (151). The ‘return’ to the use of the Henry system is also advocated by 

these authors (151), in particular they consider necessary to use the general pattern 

information of neighbouring fingers for identifications of marks (if a mark is found in 

conjunction with neighbouring marks that are highly likely to come from neighbouring 

fingers). 

A book with a part on fingerprints has been published (152). A perjury case, as well 

as a few errors are included. For the perjury, the case of New York State fingerprint 

examiners who identified marks that were lifted in the booking room as having come 

from burglary scenes between 1982 and 1992 is reported. The error in the Rick 

Jackson case is also reported, (two fingerprint examiners identified a print to 

Jackson, and two others, who were called upon by the defence, found an exclusion), 

as well as the Cowans and Mayfield cases. A large number of erroneous 

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identifications is reported in Michele Triplett’s fingerprint dictionary under “erroneous 

identifications and faulty evidence (not confirmed)” (153). Here problems with the 

LAPD fingerprint unit are mentioned, as well as the case of a fingerprint examiner 

from Seminole County, Florida (also mentioned in (152) and (189)). Only secondary 

sources are available for some of these problematic cases, for the LAPD this is the 

Los Angeles Times (154), and for the Houston crime laboratory the Houston 

Chronicle (155). Although some of these errors remain unconfirmed, recent events 

tend to show that profound problems concerning more than one case (LAPD 

laboratory and the Seminole county examiner) persist and are difficult to handle; 

therefore, appropriate reactions to such events should be foreseen. The results of the 

CTS test of 2010 (190) support this conclusion. 

A discussion of erroneous exclusions, taking into account the fact that there are 

lower thresholds applied to exclusions (a single discrepancy that cannot be 

explained) than to identifications is proposed; the author emphasizes that the way 

false negatives are viewed depends on the individual. Also, the author discusses the 

applicability of the logic of hypothesis testing, and type I and II errors, directly to 

fingerprint conclusions. He proposes to rather view ACE-V as ternary predicate logic 

(which states that an outcome of -1, 0 or +1 is possible) (156). 

In order to address the question of how different evidence types will affect jury 

verdicts, a questionnaire was given to jurors (n=233) and students (n=383); the first 

question addressed the perceived accuracy of the evidence, where fingerprints were 

rated as being slightly less accurate than DNA by both populations. The remainder of 

studies is carried out with students only. The participants received a case scenario 

(either a rape or a homicide) including different evidence types that were either 

incriminating or exculpatory. Incriminating DNA, fingerprint or victim testimony 

resulted in 100% of guilty verdicts, while hair analysis and eyewitness testimony lead 

to 92% and 75% of guilty verdicts, respectively. For the murder scenario, 

incriminating DNA evidence is the only evidence type leading to 100% of guilty 

verdicts. Hair analysis yields 83% of guilty verdicts, while fingerprint evidence leads 

to 67% of guilty verdicts. Victim testimony and eyewitness testimony lead to 55% and 

60% of guilty verdicts, respectively. The results when the evidence is exculpatory are 

particularly interesting: in the rape scenario, the percentages of guilty verdicts remain 

at 15% (DNA), 70% (hair), 67% (fingerprint), 27% (victim testimony) and 54% 

(eyewitness testimony), while these numbers are of 36% (DNA), 25% (hair), 46% 

(fingerprint), 64% (victim testimony) and 27% (eyewitness testimony) in the murder 

scenario. In this first study, only the location for DNA (and hair) was given; it was said 

that the DNA was from semen found on the victim for the rape case, under the 

victim’s fingernails and on her clothing in the murder case, while hair evidence was 

found on the victim in the rape and the murder case. A second study addressed the 

fact that this might have had an influence; this study was carried out with a student 

sample. Now, for all evidence types, the location of the evidence was given. Also, 

only a probability of guilt was asked for; the results are the following mean 

assessments of guilt : 65.2 (DNA), 55.71 (blood type), 48.00 (fingerprint), 43.15 (hair 

fibre) and 36.74 (eyewitness testimony). From a verification question, which asked 

whether each evidence type matched, it appeared that a part of the sample where 

only blood type was given as incriminating evidence considered DNA to be matching. 

In the third study reported, different elements related only to DNA evidence were 

239

tested (whether pre-trial beliefs regarding DNA, the reliability of the laboratory, or the 

style of cross-examination influenced the verdict (157)). 

Starting from dictionary definitions of the words ‘subjective’ and ‘objective’, Leo 

develops arguments in relation to different court cases where the use of the word 

‘subjective’ was important, according to this analysis (191). 

An important change occurs in the argument about fingerprint identification: it is 

being more and more considered an opinion, based on the specialists judgement, 

rather than fact (158-160). 

Bias 

Review papers in the subject of cognitive bias are now available (26, 192, 193). A 

short summary of the research contributions made during the reviewing period is 

given hereinafter. 

Whitman and Koppl (161) discusses biasing effects of the justice system and the 

integration of forensic services with the police, as well as reaction possibilities. This 

article also discusses ‘verification shopping’, which consists in searching for 

someone who will verify the original conclusions. Dror and Rosenthal describe two 

previously published studies and carry out a meta-analysis, reuniting the effects of 

bias observed in those two studies; the effect size thus obtained is quite large and 

with a p-value of 0.015 would generally be considered to be significant (162). Bias in 

verification has been tested for both experts (n=43) and novices (n=86). Three 

groups were created for both the experts and the novices. The first group carried out 

comparisons without any information (control), the second group received the 

comparisons and had answers provided that were stated to have been given by a 

fingerprint examiner trained to competency (low bias group); in the third group, the 

answers provided were said to come from prominent and internationally recognized 

expert (high bias group). For experts, there was a significant increase in inconclusive 

opinions in the low and high bias groups with respect to the control group. This 

increase is larger for the three different source trials than for the three same source 

trials (only one of the same source trials produced inconclusive opinions). One trial 

lead to an increase in the relative frequency of experts’ conclusion towards the bias 

prompt; in this trial the ground truth was different source, and the bias prompt was 

inconclusive. For novices, there were more inconclusive conclusions for the different 

source trials than for the same source trials, but there were not more inconclusive 

opinions in the bias groups with respect to the control group. There was, for novices, 

an increase in the relative percentage of responses in agreement with the bias 

prompt in the bias groups with respect to the control group (163). 

Dror analyses the sources of biases that are represented by the four idols of Francis 

Bacon, in order to improve forensic science by identifying such vulnerabilities (164). 

A study has been carried out with 70 fingerprint examiners, in a way resembling 

verification, and declared as an experiment. Examiners were in two groups, high and 

low emotional context (an allegation of murder and one of forgery). This study did not 

find any bias effects (165). The study has been criticised concerning both 

experimental flaws and problems with the interpretation of the data (166, 167). The 

authors react to both of these criticisms (168, 169). 

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Busey and Loftus discuss bias in eyewitness testimony and fingerprint comparison 

contexts, based on past research (170). 

Charlton et al. explore the satisfaction in doing fingerprint comparisons through 

interviews with 13 experienced fingerprint examiners. In particular, they highlight the 

job satisfaction which is related to the sense of pride the examiners have with respect 

to their skills, and also satisfaction with crime solving. Finding a match is described 

as a buzz, a really good feeling, and examiners like to see cases through to 

conclusion; there is a need for closure (171). 

Discussions on validity of fingerprint comparisons and ACE-V 

A special issue of Law, Probability & Risk, which appeared in 2008, is anchored 

around the ACE-V process. After an introduction (55), the validity (interpreted as both 

validity and reliability here) of the ACE-V method is discussed. This discussion 

includes proposals to carry out testing for validity. Perhaps most importantly and 

contrary to the current evolution of evaluation procedures, they propose the use of a 

numerical standard for evaluation (without defining it), but also, alternatively, a 

‘training and experience’ standard (56). The use of a numerical standard is also 

defended in (194), as a safeguard, and to protect the infallibility of fingerprint 

evidence. The reaction by Champod (57) to the article by Haber and Haber (56) 

clarifies that ACE-V is just a protocol, which is general to the forensic comparison 

sciences ; it does not insure reliability. A discussion of the usefulness of statistical 

models, specifying the selectivity of fingerprint features, is presented, along with a 

call for transparency including the spelling out of the ACE-V protocol. Cole (58), also 

in reaction to the Haber’s article (56), denies ACE-V the status of a methodology and 

discusses the irrelevance of the scientificity of the approach, highlighting the 

importance of validation (and reliability). Mnookin (59) estimates that indeed, 

research is necessary but that fingerprints could be used in court without the actually 

unfeasible testing Haber and Haber require “so long as the fingerprint community has 

done as much as it reasonably can do to establish the validity of its approach and the 

accuracy of its practitioners’ conclusions”. She considers particularly problematic that 

such feasible research has not been carried out by the fingerprint community. Haber 

and Haber react to these articles, and the arguments presented therein (60). They 

are also the authors of a book which describes the forensic use of fingerprints, how 

fingerprint work is carried out, revises court-related issues, error rates and accuracy, 

bias, training issues and finally fingerprint testimony (172). A book review by Bono 

has just been published (195). 

Cole emphasizes that it is the validity of the process that needs to be considered 

(173). Here, the author argues that expert testimony (not expert evidence) needs to 

be thought about, and that “evidentiary claims stand along a continuum of 

trustworthiness” rather than being clearly separable into reliable and unreliable 

evidence. In particular, rather than considering admissibility of evidence, according to 

the author, attention should be paid to over- or under-claiming during testimony. He 

then analyses fingerprint testimonies of 34 case transcripts (173), focussing on the 

“source attribution moments”. These were then divided into Process Statements (the 

mark was identified/matched to the inked impression), Source Attribution Statements 

(the defendant made the print/defendant could be the only source) and Identity 

Statements (the fingerprint was one and the same with the mark). It is considered 

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that Process Statements do not really attribute the mark to a source; it is only stated 

that the two impressions match, but how often this could happen in a population is 

not assessed. Source Attribution Statements are considered to represent overclaiming, 

while Identity Statements, which were less frequent in the 34 cases 

reviewed, go against the principle that no two impressions could be identical. In a 

later article, the same author argues that fingerprint individualization evidence 

satisfies neither the Daubert nor the Frye admissibility standard (196). Mnookin (174) 

argues that testing is needed to assess the scientific validity of a method, rather than 

explanations of the premises and the methods used. She considers that to know how 

a method works is less important than knowing that it works. An argument against 

the usefulness of uniqueness and the lack of necessity for individualisation 

conclusions in the legal system is presented in (175). A different approach to 

uniqueness, individualization and identification is used by Kaye (181), who discusses 

general and special uniqueness as well as ‘local’ and ‘universal’ individualizations. 

The same author argues that very small probabilities of fortuitous correspondence 

can reasonably be considered negligible (182). The author links this argument to the 

way that all science works today: Some inductions which, according to both laws of 

probability and Popper's writings, cannot be absolutely proven as true are, 

necessarily, considered as such. This directly responds to Saks and Koehler, who 

criticise the “uniqueness fallacy” which can occur when the frequency of occurrence 

is smaller than the number of objects that exist (e.g. f*n

Furthermore, the authors propose to consider the premises of fingerprint 

individualisation (persistence, unicity and classifiability) as a theory, hypotheses that 

have not been falsified through testing to date and therefore, being supported by 

evidence, have gained the status of theory (184). ACE-V, as applied by fingerprint 

examiners in a laboratory, has been tested in order to assess the accuracy, 

precision, reproducibility, repeatability and biasability of conclusions. Overall, three 

erroneous identifications (out of 720 comparisons) were observed but were attributed 

to transcription errors. Two of these were submitted in the course of the study to 

verification and were not verified, while the third erroneous identification was resubmitted 

to the same analyst, who did not repeat the error. Erroneous exclusions 

were more prevalent and less likely to be caught in verification. Also, divergences 

appeared between definitive exclusions (identification/exclusion) and inconclusive / 

no value conclusions. In particular, some comparisons resulted in ‘identification’ 

decisions from some examiners, and in ‘inconclusive’ decisions from others (185). 

Accuracy in fingerprint comparison has been tested using laypeople (186) ; it is 

shown that some fingers are more easily discriminated than others. 

Whether written training programs exist, whether they adhere to SWGFAST 

guidelines, whether the interpretation of these guidelines is consistent, and whether a 

difference exists between ASCLD/LAB accredited agencies and non-accredited 

agencies concerning the training standards has been investigated by the means of a 

survey (187). Out of 168 surveys sent out to members selected from the 2007 

International Association for Identification directory, 75 were returned. 75% of the 

respondents claimed having a written training program, and 72% claimed adhering to 

SWGFAST guidelines. Five follow-up questions were used to determine whether 

these agencies actually did follow the guidelines, and, based on these follow-up 

questions, only 25% do. As to the consensus in the interpretation of the guidelines, it 

exists in 7 out of 11 categories tested; the four categories where no consensus was 

apparent are degree requirements, the duration of the training program and of 

supervised case review, and the existence of a written pass or fail policy. There was 

a significant difference between accredited and non-accredited laboratories on a 

majority of the points examined, and the hypothesis that there is no difference 

between these two types of laboratories was therefore rejected. 

The shortcomings of current proficiency tests for estimating error rates and a way to 

obtain error rates for fingerprints through proficiency tests is discussed in (188). 

An argument for several verifications, including an economic perspective has also 

been presented (189). 

2.2 Fingermark composition, detection and visualization 

This chapter is structured as follows: the first section is dedicated to the composition 

and evolution of the fingermark residue under various environmental conditions. It is 

followed by the presentation of the recent developments in fingermark detection, the 

scientific contributions being classified according to the technique, the context, or the 

substrates. 

243

The following contributions constitute recent reviews that can serve as good starting 

points for readers not accustomed to the range of methods available in the field: 

detection of latent fingermarks (197-199), forensic chemistry (200), and forensic 

science (1). 

When summarizing the experimental studies published these last three years, we 

chose to focus on the “fingermark sampling” protocols, if described by the authors. 

This encompasses the nature of the secretions that were used (i.e., natural/nonenriched, 

eccrine-rich, or sebum-rich), the total number of donors and fingermarks, 

the time between the deposition of the fingermarks and the processing of the 

samples, and if depletion series were considered. A great versatility has been 

observed among the contributors, with protocols close or far-off realistic conditions 

(not necessarily casework-like). This emphasizes the fact that the sampling protocol 

is a key-element to be aptly chosen since it will influence the possibility to compare 

experimental results between researchers and to operationally implement a new 

technique. In this context, the recent publication by Terry Kent may constitute a step 

forward in the standardization of the sampling protocols (201). In this contribution, 

some sampling protocols and testing methods were discussed (e.g., effect of 

grooming, use of split fingermarks and of depletion series, timescale, substrates, 

donors, and performance evaluation). Another study, proposed by Croxton et al., 

could provide a quantitative proof of the lipid over-representation in “groomed” 

fingermarks compared to natural ones (202). In a third study, an original concept has 

been presented to empirically quantify the differences in contrast between the ridges 

(of a fingermark) and the valleys (generally: the underlying substrate) (203). The socalled 

“relative contrast index” could help in assessing the actual efficiency of a 

tested technique, and simultaneously provide a documentable and objective way of 

measuring contrast enhancement that could be used to perform comparative studies. 

In this context, Vanderwee et al. performed a literature survey of ca. 45 published 

papers and reported the information that could be found in the manuscripts in terms 

of visual assessment, scoring protocols, side-by-side comparison, or development 

time (non-exhaustive list of reported parameters) (204). It would be interesting to see 

if the forensic science research community will follow these advices and propositions 

in the coming years. 

2.2.1 Composition, aging, and persistence of fingermarks 

Attempts to correlate the chemical composition of latent fingermarks with 

the age of the individuals who have left them were published (205, 206), 

both concluding that it is possible to estimate the age of an unknown 

individual by analyzing the infrared spectra of marks. In another study, the 

analysis of the amino acid composition of latent fingermarks was used as 

a witness of a genetic disease (207). Finally, the composition in amino 

acids and lipids between natural and sebum-rich fingermarks was 

compared using GC-MS (202). 

The persistence of fingermarks under wet conditions has been 

investigated by various authors (208-210). If all emphasized the 

successful processing of wetted substrates to detect latent fingermarks, 

their conclusions differed about the techniques to be applied in such 

situation. Finally, photo- and thermal-degradation of eccrine secretion 

components has been studied (211). 

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Fourier transform infrared spectroscopy (FTIR) was used to determine if changes in 

fingermark composition could be correlated with the age of the person that has left 

the mark (205). For this study, latent fingermarks deposited on tin plates by 78 

individuals aging from four to 68 years were considered. Analyses were performed 

24 hours after deposition for children (0-19 years), and 48 hours after deposition for 

adults (20-70 years). The results showed that a combination of constituents could be 

found, whose composition linearly changes with age, with a significant shift around 

puberty. A linear model of age as a function of the FTIR spectrum has been 

proposed, with a root mean square error of calibration of less than four years. This 

linear model shows an inflection at the age of 25. If two linear models were 

constructed (i.e., for people

In the context of standardization of the fingermark sampling protocols, the study 

proposed by Croxton et al. showed that lipids are logically over represented in 

“groomed” fingermarks (i.e., when the donors are asked to rub their fingers on their 

face or hair prior the deposition of fingermarks) compared to natural marks (202). 

Eighteen donors were asked to leave “natural” fingermarks and sebum-rich ones on 

a non-porous substrate. All the fingermarks were immediately analysed using gas 

chromatography coupled with a mass spectrometer (GC-MS). On average, fatty acid 

content could be over represented by a ratio of 4:1, squalene by a ratio of 16:1, and 

the total sebaceous mass by a ratio of 6:1, compared to natural marks. This over 

representation in lipids could seriously compromise the validity of some published 

results. It should be noted that the amino acid composition remained significantly 

similar between the two kinds of fingermarks. Moreover, the relative abundance of 

amino acids was consistent with previously published studies (212), with serine as 

the most abundant amino acid followed by glycine, alanine and aspartic acid. Finally, 

no correlation between the total amount of fatty acids and of amino acids was 

observed in both kinds of fingermarks. This result seems logical since these 

chemicals are secreted by different glands. 

Soltyszewski et al. studied the effect of water on the recovery of fingermarks and 

DNA profiling (209). For this study, 456 sebum-rich fingermarks were left by eight 

donors, on glass (10 seconds of contact time, medium pressure). Additionally, four 

different kinds of water (i.e., river, sea, tap, and distilled), two temperature conditions 

(i.e., 5 and 20°C), and six immersion times (from one day to 42 days) were 

considered. The samples were then dried, processed by three fingermark detection 

techniques (i.e., aluminium and ferromagnetic powders, and cyanoacrylate), then 

swabbed for DNA profiling. Fingermarks were successfully retrieved for items being 

submerged in water for up to six weeks, with high success rates for one to seven 

days of immersion. Ferromagnetic powder and cyanoacrylate fuming showed to be 

more effective than aluminium powder. The quality of the fingermarks nevertheless 

decreased as the immersion times increased, and as the water temperature rose 

from 5°C to 20°C. Finally, no DNA profile could be obtained regardless of the tested 

conditions. 

Kerr et al. investigated the effect of water immersion on the efficiency of the 

commonly used fingermark detection techniques (208). For this study, non-enriched 

fingermarks were left on glass before being immersed in different water baths (i.e., 

cold, 50°C, or soapy) for varying times (i.e., five seconds and one minute). It should 

be noted that the fingermark aging time (i.e., the time left between the deposition of 

the marks and their wetting) has not been specified by the authors. We assume that 

the freshly deposited marks were immediately rinsed after their deposition. Nine 

fingermark detection techniques were compared (i.e., aluminium and gold powders, 

small particle reagent – SPR, eosin blue, erythrosine B, physical developer - PD, 

gentian violet, sudan black, and cyanoacrylate fuming) according to their ability to 

reproduce ridge details. As a conclusion, the authors recommend to first air dry 

wetted glass samples vertically, before processing them with either aluminium or gold 

powder. If vertical drying is impossible, they recommend using SPR while the 

samples are still wet. It should be noted that cyanoacrylate fuming gave poor results 

on the dried samples, which is in contradiction with the above-described study (209). 

246

Wood & James investigated the persistence of latent fingermarks left on white paper 

and uPVC plastic immersed in different kinds of water (i.e., river, rain, and tap) or in 

an accelerant (i.e., unleaded petrol) (210). For this study, freshly-deposited sebumrich 

fingermarks from one donor were used. The items were immersed in the different 

liquids for varying times (i.e., one hour, 24 hours and one week) before being dried 

and processed with commonly used fingermark detection techniques (i.e., magnetic 

powder, iodine fuming, ninhydrin, cyanoacrylate, SPR, PD, oil red O - ORO). On 

paper, the latent fingermarks were found to persist for over a week when immersed 

in water with successful application of magnetic powder, PD and ORO. After one 

week, identifiable marks were obtained only with ORO. Logically, ninhydrin gave poor 

results regardless of the immersion times due to the amino acids solubility in water. A 

more pronounced detrimental effect has been observed for the items immersed in the 

accelerant, with identifiable marks detected only for a 24-hour immersion time, using 

ninhydrin. On plastic, marks were easily detected after a water immersion for over a 

week, using all the compared techniques (with the notable exception of iodine 

fuming). The detrimental effect of the accelerant has been less than for the paper, 

with successful detection for all immersion times and techniques (except iodine 

fuming). Among the tested techniques, the authors recommend the use of ORO and 

SPR to detect fingermarks on porous and non-porous substrates, respectively. 

To study the effect of sunlight exposure and temperature on eccrine secretions, 

aqueous standard solutions of aspartic acid, glutamic acid, glycine, histidine, 

ornithine, serine, threonine, urea, and lactic acid were prepared (211). The photodegradation 

study was performed as follows: the standards were deposited on Teflon 

disks and steel coupons, then allowed to dry overnight in the dark before being 

exposed to a 200–500 Watt Xe⁄HgXe arc lamp for varying times (up to ca. 3h40), 

corresponding to sunlight exposure times ranging from 0 to 56 days. The thermaldegradation 

study was performed by depositing the standards on galvanized steel, 

then allowing them to dry overnight in the dark before being exposed to temperatures 

ranging from 50 to 150°C using a heat gun. Mass spectrometry was used to analyse 

the samples afterwards. No notable photo-degradation of the amino acids and urea 

was observed for both substrates, even after 56 days of sunlight exposure. Lactic 

acid however quickly underwent degradation (already after two days), leading to the 

generation of pyruvic acid which subsequently also underwent degradation. A 

substantial thermal degradation of the amino acids and urea occurred for 

temperatures above 100°C. For lactic acid, the thermal degradation started between 

50 and 100°C (along with pyruvic acid generation and subsequent degradation). 

2.2.2 Amino acid reagents 

The use of amino acid sensitive reagents still constitutes the method of 

choice to detect latent fingermarks on porous substrates. Even if such 

reagents have been known for a long time (199) and are widely used in 

casework, two major contributions have somewhat renewed the subject. 

First, the development of an amino acid standard obtained by printing a 

pattern using a modified inkjet printer (213), and a collaborative test 

carried out using this standard (214). Second, a thorough study detailing 

the intimate mechanisms of the reaction of amino acids with ninhydrin, 

1,8-diazafluoren-9-one (DFO) and 1,2-indanedione/zinc (215). Besides 

these contributions, the use of promising ninhydrin analogues has been 

247

investigated (216-218), as well as the effect of pressure and sweat on the 

quality of ninhydrin-processed fingermarks (219). An alternative to the use 

of a temperature- and humidity-controlled development cupboard has also 

been proposed (220). Concerning DFO, Schwarz and Beisel proposed a 

new formulation, said to be more robust against the formation of two 

phases (221). The studies on 1,2-indanedione were mainly focused on the 

optimization of the formulations and of the development procedures (222- 

224). Finally, lawsone (225) and isatin (226) were proposed as new 

promising amino acid reagents, while pDMAC was proved to actually react 

with amino acids, and not with urea as it was thought for long (227). 

Ninhydrin and analogues 

A computational study, initiated by Petraco et al. (see Interpol report 2004-2007) on 

the reaction mechanisms between ninhydrin (and analogues) and amino acids, has 

been pursued (216). According to their conclusions, some analogues could constitute 

promising substitutes for ninhydrin and should be synthesized for experimental tests. 

Following their research on a one-stage “ninhydrin / metal salt” premix formulation 

behaving like a dual reagent (i.e., leading to both coloured and luminescent 

fingermarks), Almog et al. showed that the use of commercially available ninhydrin 

analogues led to better results (217). By mixing 5-methoxyninhydrin (MN) or 5methylthioninhydrin 

(MTN) with zinc or cadmium salts, coloured fingermarks could be 

obtained (MN: orange with zinc, and pink with cadmium; MTN: pink with zinc, and red 

with cadmium) and observed in luminescence at room temperature, without the use 

of liquid nitrogen. The “MTN ⁄ ZnCl2” gave the best results, with fluorescence intensity 

comparable to the one of 1,8-diazafluoren-9-one (DFO), but with a much stronger 

visible colour. If classified according to the colour, the different reagents were ranked 

as follows: MTN/ZnCl2 ≅ MN/ZnCl2 > Ninhydrin/ZnCl2 > DFO. If the classification is 

made according to the fluorescence intensity at room temperature: MTN/ZnCl2 = 

DFO ≥ MN/ZnCl2 >> Ninhydrin/ZnCl2. 

Jasuja et al. investigated the effect of various parameters that occur during the 

deposition of a fingermark on the efficiency of the subsequent ninhydrin process 

(219). For this study, thirty individuals (among which five were considered as good 

sweat donors, and five as bad sweat donors) aged from 20 to 25 years left latent 

eccrine marks on white paper following different conditions: conventional situation 

(no specific pressure, no induced sweat), with pressure (i.e., 50 g, 100 g, 150 g, and 

200 g), and with induced sweat production before deposition using a polyethylene 

bag around the hand (i.e., for 30 to 180 seconds). Less than 24 hours later, all the 

fingermarks were processed using ninhydrin. Linear relations were established 

between the quality of the developed marks and the pressure intensity, under the 

studied conditions. As the induced sweating time increases, the quality of the marks 

increased as well for all types of donors (mixed, good and bad donors). An induced 

sweating time of 120 seconds produced the same quality of development of the 

fingermarks, irrespective of the donor quality. In case of good sweat donors, 

smudging of the fingermarks could occur when the pressure applied and induced 

sweating time exceeded 200 g and 180 seconds, respectively. On the contrary, in the 

case of the bad donors, greater pressure and induced sweating times definitely 

resulted in marks of better quality. 

248

The successful modification of a commercial bubble jet printer so that the cartridges 

are filled with amino acid standard solutions has been proposed as an alternative to 

the deposition of fingermarks from fingertips (213). By doing so, one gets rid of the 

variability induced by actual fingermarks when comparing two techniques (even if 

split fingermarks are considered). The pattern to be printed contains a scanned 

fingerprint and halftone printed gray scale fields. This method is quite reproducible 

with a standard deviation of 7% calculated after having printed 126 patterns using 21 

different cartridges (the method of calculation relies on the weight of the cartridges 

before and after the printing, so that the quantity of deposited amino acids can be 

evaluated). This method will not completely replace actual fingermarks (that should 

be considered before any casework application) but could be useful at the beginning 

of the comparison process between different detection techniques. A collaborative 

test has been carried out using this method to assess the efficiency of ninhydrin and 

DFO processes as used in different laboratories (214). Concerning ninhydrin, the 

results emphasized the role played by humidity during the development on the 

sensitivity of the method. 

A thorough mechanistic study has been conducted on DFO, ninhydrin, and 1,2indanedione 

(with and without zinc chloride) (215). It has been known for long that 

these reagents may produce varied results in different geographic areas, on different 

paper substrates, or under different environmental conditions, but no explanation of 

such different behaviours was actually reported. For this study, aqueous solutions of 

the nine major amino acids contained in a secretion residue (i.e., serine, glycine, 

ornithine, alanine, threonine, histidine, valine, leucine, and lysine) were deposited as 

spots on three kinds of porous substrates (i.e., ashless filter paper, 10% recycled 

copy paper, and cellulose-coated TLC plates), before being processed by the abovecited 

techniques. Absorption and luminescence spectra were finally recorded and 

analyzed. One of the most important observation made by the authors is the role 

played by water (moisture) in the successful reaction of 1,2-indanedione with amino 

acids. The role of zinc chloride has also been investigated as it seems to act as a 

catalyst, bypassing the need for high level of moisture for 1,2-indanedione to react. 

Almog et al. are currently working on analogues for ninhydrin and 1,2-indanedione, 

bearing thiol groups (-SH) in their chemical structure (218). The aim is to artificially 

enrich latent residue with thiols so that they could constitute efficient anchoring sites 

for metal nanoparticles. This research is still in its preliminary stage, but it illustrates 

quite well the current trend consisting in using nanoparticles as new efficient 

reagents to detect fingermarks (as further developed in section 2.2.6). 

Schwarz & Hermanowski proposed an alternative to the use of a development 

cupboard (with controlled levels of humidity and temperature) to develop ninhydrinprocessed 

items (220). The use of a saturated aqueous salt solution in place of water 

vapors could help developing fingermarks even if the item is left in a classical bench 

cupboard, at room temperature. They observed that after two or three days, the 

development was optimal with no background staining. A prolonged exposition time 

could lead to some unwanted background staining, according to the nature of the salt 

(e.g., sodium chloride or potassium citrate). 

249

1,8-diazafluoren-9-one (DFO) 

Schwarz and Beisel noticed that different DFO solutions prepared according to the 

same formulation showed different propensities to form two phases, and secondly, 

that unstable solutions lead to inferior detection results (221). A more robust 

formulation containing only HFE-7100, acetic acid, ethyl acetate, methanol and DFO 

(as a prerequisite) was therefore proposed. Petroleum ether, chloroform and 

dichloromethane were excluded, because of health and safety issues. The polarity of 

the solution is sufficiently important to avoid the formation of two phases, but is not 

sufficiently important to make inks run. The following development conditions were 

recommended: two immersions with a drying time in between, and then 40 minutes 

at 110°C. 

1,2-Indanedione 

The use of 1,2-indanedione as a competitive replacement of DFO (and ninhydrin) 

has been seriously considered after the publication of Stoilovic et al., who proposed a 

one-stage premix formulation containing ZnCl2 (Ind-Zn) (228). This formulation 

seems now to be widely accepted. Only few works were published since 2007, 

consisting mainly in minor optimizations of the formulation to enhance both colour 

and luminescence of the developed fingermarks. It has indeed been noticed by 

various people that 1,2-indanedione may perform inconsistently among laboratories 

worldwide. 

In an attempt to enhance the results obtained with the formulation published by 

Stoilovic et al.(228), Russel et al. proposed to increase the amount of zinc chloride, 

to dip the documents twice in the solution, and to increase the heating time from 10 

seconds to 30 seconds (222). Bicknell & Ramotowski performed a thorough study 

about the effect of laboratory humidity (% RH), moisture content of the paper, 

temperature, and zinc chloride concentration (223). They showed that there exists a 

correlation between the moisture content of the paper on which the prints are 

deposited and the % RH of the air, having for consequence that the development 

conditions have to be adapted accordingly. They also observed that increasing the 

quantity of ZnCl2 improved the results up to a maximum limit above which the 

efficiency begins to decrease. Finally, they showed that Ind-Zn was superior in colour 

and fluorescence intensity compared to DFO, even for 11-year-old marks. 

If several studies illustrated the superiority of Ind-Zn compared to DFO, it is not the 

case with the comparative study performed by Sears et al. in the United Kingdom 

(224). When comparing various formulations of Ind-Zn with the DFO formulation 

recommended by the Home Office (HOSDB), they concluded that DFO develops 

more fluorescent fingermarks of high quality than does the Ind-Zn solution. This was 

verified on a majority of the tested substrates, especially on two-day-old fingermarks. 

Nevertheless, the difference in luminescence tended to equalize (even to reverse) as 

the marks aged (14-day-old ones). It should be noted that, similarly to the 

observations made by the other authors, increasing the amount of ZnCl2 in the 

working formulation has a positive effect on the quality of the detected fingermarks. 

250

Other reagents 

Lawsone (2-hydroxy-1,4-naphthoquinone) is a new amino acid reagent that has been 

proposed by Jelly et al. to detect fingermarks on porous substrates (225). The 

reaction of lawsone with latent secretions leads to purple-brown marks that are also 

luminescent. However, only preliminary results on filter paper were presented, and 

further research is required to assess the quality of this reagent compared with 1,2indanedione 

or DFO. 

Isatin (1H-indole-2,3-dione) is structurally similar to ninhydrin or 1,2-indanedione and 

has consequently been proposed as a new amino acid reagent able to detect latent 

fingermarks (226). For this study, six donors were asked to leave fingermarks on 

white paper. The marks were aged during two days before being processed. The 

composition of the working solution, the development conditions, and the use of 

metal salts (i.e. zinc acetate, zinc chloride, and zinc nitrate) in the working solution or 

as post-treatments were studied. The authors concluded that a 0.05% (w/v) of isatin 

in 12.5% (v/v) acetone, 87.5% (v/v) dioxane and carbonate buffer gave the most 

intense luminescence, but with a less intensely coloured product. The development 

conditions were similar to those recommended for 1,2-indanedione, the samples 

being heated at 180°C for 10 s. The samples could subsequently be post-treated by 

dipping them in an ethanolic zinc chloride solution, and reheating them at 180°C for 

10 s. When compared with DFO and 1,2-indanedione/zinc on half-cut fingermarks, 

isatin consistently showed lower luminescence intensity and greater unwanted 

background luminescence. When compared with ninhydrin, isatin consistently gave 

no coloured fingermarks. These results illustrate the fact that further research is still 

required before proposing isatin as an alternative to the classical amino acid 

reagents. 

Para-dimethylaminocinnamaldehyde (pDMAC) has for long been considered as a 

reagent able to detect latent fingermarks by targeting urea contained in the 

secretions. A recent study performed by Lee et al. tends to prove that pDMAC is not 

reacting with urea but rather with amino acids (227). When compared with classical 

amino acid reagents to detect fingermarks on thermal papers, such as ninhydrin and 

DFO, pDMAC was shown to be far less effective in terms of colour and 

luminescence. Consequently, its use is not recommended. 

2.2.3 Cyanoacrylate fuming 

The possibility to rejuvenate old fingermarks to improve the cyanoacrylate 

fuming process has been investigated, and led to the conclusions that the 

items should be exposed to either acetic acid (229) or methylamine 

vapours (230). The role played by the environmental pH on the 

morphology and length of the polymer chains has also been investigated 

(231). An attempt to optimize the fuming time when processing 

fingermarks on skin failed in determining optimal conditions (232). Finally, 

a detrimental effect of cyanoacrylate fuming has been observed if the 

items (latex/nitrile gloves or duct tapes) have to be subsequently 

processed by powder suspensions (233). 

251

Wargacki et al. proposed two studies about the understanding of the cyanoacrylate 

fuming mechanisms. Using mass accumulation and molecular weight analysis, they 

first showed that carboxylate moieties (coming from lactic acid or amino acids) seem 

to be the primary initiator of the polymerization process and constitute better initiators 

than primary amines (231). They also showed that the influence of the pH occurs at 

the latter stages of the polymerization process (not in the initiating step). Indeed, a 

basic environment will inhibit the polymerization chain termination, leading to the 

formation of long chains with high molecular weight at long exposure times, while an 

acidic environment will promote the termination process and the formation of shortchained 

polymers. They also showed that, contrary to what is believed, water does 

not constitute the primary initiator and does not create the long-chained polymers 

that are visible on a fumed fingermark. In another study, Wargacki et al. studied the 

aging process and the possibility to rejuvenate aged fingermarks (229). The 

decreased quality observed for aged fingermarks seems to be due to a loss of 

initiators as a consequence of the erosion and degradation processes induced by the 

loss of water. Exposing latent eccrine fingermarks to vapors of acetic acid (to provide 

polymerization initiators) or ammonia (to bring the pH to basic values) for several 

minutes constitute two efficient enhancement processes able to rejuvenate old 

fingermarks and improve the growth of polymers on the ridges. Acetic acid is 

somewhat preferred compared to ammonia, given its higher efficiency and its lower 

toxicity. 

Similarly to Wargacki et al., McLaren et al. tried to rejuvenate old fingermarks to 

improve the quality of development once fumed with cyanoacrylate (230). They 

observed a minor positive effect due to the use of acetic acid and water vapours, 

whereas a more consistent effect was observed with ammonia. They nevertheless 

noticed that the donor plays a great role in the rejuvenating capabilities when using 

these chemicals. As an alternative, they recommend the exposition of old/dry 

fingermarks to vapours of a 10% w/v aqueous methylamine solution, for one hour. In 

this case, the role played by the donor seems not to be as pronounced as that of 

ammonia and acetic acid. It should be noted that, contrary to acetic acid and 

ammonia, some cases of excessive polymer formation across the ridges were 

observed with methylamine (leading to a degradation of the mark instead of an 

improvement). Additionally, health and safety issues were identified and should be 

investigated. 

King studied the effect of the fuming time when trying to recover latent fingermarks 

on skin using cyanoacrylate (232). This question arose from contradictory 

observations published in the literature, in which optimal fuming times can vary from 

a few minutes to more than an hour. Using pig skin (previously washed, dried, and 

warmed at room temperature) on which fresh and sebaceous latent fingermarks from 

a unique donor were deposited, King set the fuming times between 10 minutes to 

125 minutes. It should be noted that the fuming chamber was quite rudimentary, with 

no control of humidity or of the heating temperature. Once fumed, the fingermarks 

were powdered using a black magnetic powder and their quality rated. The author 

failed to find a significant relationship between the quality of the developed 

fingermarks and the fuming times. 

The influence of cyanoacrylate fuming prior to a subsequent processing of tapes and 

gloves with powder suspension has been investigated (233). For this study, one 

252

donor was asked to deposit fingermarks on four kinds of tapes (i.e., red and gray 

duct tapes, masking tape, and Scotch tape), two donors wore two kinds of gloves 

(i.e., latex and nitrile) for a variable amount of time (not specified in the article) before 

removing them, and worn nitrile gloves were processed after having been removed 

from trash (unknown donors). One hour after fingermark deposition, the samples 

were either processed by the sequence “cyanoacrylate fuming – black powder 

suspension (BPS)”, either by BPS only. A detrimental effect caused by cyanoacrylate 

fuming was observed, since no ridge details could be obtained by BPS if the items 

were first processed with cyanoacrylate. 

2.2.4 Oil red O (ORO) & Physical developer (PD) 

When considering wetted porous items, PD and ORO constitute the two 

techniques of choice available to the forensic scientists. A comparison 

study between ORO and PD led to the conclusion that ORO performs 

better on fresh marks (up to four weeks) and PD on aged ones (more than 

one month) (234). Another study showed that ORO outperforms PD on 

sebum-rich marks, but that both techniques perform equally on natural 

marks (235). The following processing sequence was recommended: 

“ORO first, followed by PD”. In another study, a commercial PD 

formulation was shown to be as efficient as a “home-made” one (236). 

Oil red O is quite a recent technique (first proposed in 2004) based on a lysochrome, 

staining the lipid fraction of latent secretion. This technique is usually proposed in 

competition (or in complement) with PD to detect fingermarks on porous substrates, 

especially if they have been wetted. Salama et al. performed an extensive 

comparison between both reagents, on various porous substrates (234). They 

concluded that both techniques are complementary, with the age of the latent 

fingermarks playing a key role. ORO is efficient on fresh fingermarks up to four 

weeks. For older marks, PD is preferred. Since it is generally difficult to assess the 

age of the marks to be detected, it is thus recommended to apply these techniques in 

sequence. On wet substrates, PD followed by ORO gave excellent results. 

Nevertheless, it is recommended to apply ORO first, followed by PD. This sequence 

is governed by the fact that PD could severely deteriorate a substrate on some 

occasions (e.g., blackening of the surface), annihilating subsequent attempts to 

process the item for fingermarks. 

When comparing ORO with PD, Wood and James studied the role played by the 

substrates and the liquid into which they are immersed (235). For this study, one 

donor was asked to deposit two kinds of fingermarks (i.e., natural/ungroomed ones 

and sebum-rich ones) on four different papers (i.e., standard print, high-quality print, 

lined and card). The marks were then left for one hour before being immersed in four 

different liquids (i.e., tap, rain and river water, and un-leaded petrol) for times ranging 

from one hour to one week. After the immersion, the samples were dried and cut in 

half, each half being processed either by ORO or by PD. Both techniques were 

unable to detect any mark on the samples immersed in the petrol, for all the 

conditions tested (certainly due to solubilisation of the fingermark lipid components). 

When the samples were immersed in water, ORO outperformed PD for all immersion 

times and paper types, when loaded marks were to be detected. In this case, after 

253

one day of immersion, PD failed in detecting identifiable marks (contrarily to ORO). 

For natural marks, both techniques performed equally (in few cases, PD even 

developed better marks than ORO). This could be explained by the fact that PD 

targets more constituents than ORO (which only stains lipids). Finally, no difference 

was observed between the three kinds of water that were used. Additionally, the role 

played by the paper type is minor, at the exception of the card for which only natural 

marks of low quality were obtained. 

The efficiency of a commercially available PD processing kit (brand: Sirchie 

Fingerprint Laboratories) and a “home-made” PD solution have been compared on 

different porous substrates (236). It has been concluded that both the commercial 

and the non-commercial solutions were competitive and acting equally in terms of 

fingermark quality (contrast and clarity), the commercial formulation being somewhat 

more sensitive when less secretions are to be detected. Nevertheless, a drawback of 

the commercial solution could arise if one wants to enhance the results using 

potassium iodide (KI) to darken the background (if it contains starch) while lightening 

the marks. Indeed, such post-treatment must be photographed while the substrate is 

submerged in the solution. Given that the tested commercial PD solution was 

opaque, the resulting contrast was considerably reduced. 

2.2.5 Dry micron-sized powders and powder suspensions 

Some contributions were dedicated to the traditional, dry and micron-sized 

powders. It is possible to cite the works performed on skin (237) and fruits 

or vegetables (238), as well as the propositions of new powders based on 

phosphorescent compounds (239), on banana peel activated carbon 

(240), or on dye-doped silicate clay powder (241). Most of the publications 

were dedicated to powder suspensions (SPRs), used to detect marks on 

wetted non-porous substrates. Comparison studies were conducted 

between commercially-available SPRs (242-245), as well as between SPR 

and vacuum metal deposition (246). Original research led to the 

proposition of a luminescent SPR (247), and the relation between the 

smoothness of a surface and its processing using SPR has been 

investigated (248). Finally, it was observed that wiping an SPR-processed 

surface with a paper towel makes new marks appear (249). 

Trapecar et al. illustrated the efficiency of some fingerprint powders (especially the 

Swedish Black powder) to successfully detect fingermarks on skin (237) and fruits or 

vegetables (238), compared to chemical techniques such as cyanoacrylate fuming 

and ruthenium tetroxyde. For both studies, non-enriched fingermarks from several 

donors were left by prolonged contact with the items (several seconds), and were 

aged from 15 to 45 minutes for living bodies, from 15 to 180 minutes for dead bodies, 

and from hours to two days for fruits and vegetables, before being processed. 

Liu et al. proposed a new phosphorescent powder based on a europium-doped 

strontium aluminate. This phosphorescent powder could be of a first interest in the 

case of fluorescent or multicoloured substrates that generally lead to an unwanted 

decrease of the resulting contrast when using traditional luminescent powders (239). 

For this study, fresh and aged fingermarks (up to two-month-old marks) from different 

254

donors were left on various substrates (i.e., non-porous, semi-porous and porous). 

The advantages are various: strong phosphor / afterglow effect that lasts longer than 

the fluorescence of the substrate, no specific device needed since the substrates are 

excited under UV light for two minutes before being observed in the dark, efficiency 

on non-porous and semi-porous surfaces (as well as some porous ones, such as 

wood or fabric) as well as on cyanoacrylate fumed fingermarks, and possibility to be 

lifted. According to the authors, there are no toxicity issues with this kind of powder. 

Nevertheless, since the samples have to be irradiated under long UV for two 

minutes, detrimental effects on DNA could occur. 

Mopoung & Thongcharoen proposed a new powder to be dusted on latent 

fingermarks, based of banana peel activated carbon mixed with sodium acetate 

(20%), mineral oil (2%), and methylene blue (0.2%) (240). When compared with a 

commercially-available black carbon powder, the authors observed that the 

commercial powder adhered better on the ridges, but that background staining was 

less when using the banana peel-based powder. For this study, very fresh, sebumrich 

fingermarks were left by one donor on glass. 

Chen et al. incorporated cationic dyes (i.e., rhodamine 6G and methylene blue) into 

silicate clay powder that was subsequently rendered hydrophobic by chemical 

functionalization (241). The clay was eventually ground, to be dusted on fresh latent 

fingermarks left on glass slides. This preliminary study led to good results in terms of 

ridge details and little background staining. The authors assumed that better results 

could be obtained if the powder size distribution was optimized (for example, by 

grinding it more finely, to microscaled sizes or below), as well as the dye-loading 

step. Additionally, the attachment of biolabeling agents may help in targeting some 

specific components of the latent secretion. 

Powder suspensions (or small particle reagents – SPRs) remain one of the well 

adapted solutions for non-porous substrates that have been wetted. In this domain, 

Jasuja et al. proposed an alternative to the classical black and white SPRs by 

combining zinc carbonate (white) with different kinds of luminescent dyes (i.e., 

rhodamine B, rhodamine 6G, acridine orange, anthracene, cyano blue, and basic 

yellow) (247). Excellent results were obtained when using rhodamine 6G, rhodamine 

B and cyano blue (other dyes led to non-luminescent marks) and after 30 to 60 

seconds immersion in the working solution. It should however be noted that almost 

ideal conditions were tested with fresh marks (detected immediately after deposition, 

or after 24 to 96 hours immersion in water) on smooth non-porous surfaces (e.g., 

glass, aluminium, polyethylene). Rhodamine 6G and B being classified as suspected 

carcinogens, the cyano blue-based SPR is recommended. 

Cohen & Cohen reported quite an original method to improve the number of latent 

marks that could be detected on smooth non-porous surfaces (e.g., metal, glass, 

plastic, aluminium) after application of a molybdenum sulphide-based SPR (MoS2) 

(249). According to the authors, a thorough wiping of the surface using paper towel 

after the SPR treatment, initially done to remove dried SPR residues, led to the 

visualisation of several unobserved fingermarks. As a consequence of the wiping, the 

background turned yellowish-brown while the marks appeared as grey patterns. 

Three major observations were also made: (1) The stronger the wiping, the better the 

marks, (2) sebum-rich marks give better results than eccrine-rich ones, and (3) if the 

255

same paper towel is used on an un-treated surface, new fingermarks can readily be 

detected. The lifting of the marks was however impossible and all marks have to be 

photographed in situ. Further research is underway to elucidate the chemical and 

physical underlying mechanisms responsible for this phenomenon. 

Two commercially available white powder suspensions (WPS) were tested (i.e., 

Wetwop and Wet Powder) in comparison with a “home-made” titanium dioxide-based 

formula (242, 243). Tests were performed with depleted series of one-day-old 

fingermarks left on some non-porous surfaces (e.g., various coloured plastic bags 

and black plastic-based car cowlings), before being immersed for six hours in water, 

dried (from two to eight days), and eventually processed. The results showed that all 

three WPS formulations led to similar results, with a slight preference for Wetwop for 

the least amount of background staining left after rinsing. On the same basis, the 

authors further compared the Wetwop SPR formulation with vacuum metal deposition 

(VMD) to detect fingermarks on wetted non-porous surfaces (246). The same kinds 

of substrates were used, and the fingermark deposition procedure was kept 

unchanged (except that drying times were of two to 28 days). Fingermarks were cut 

in halves and processed separately with the two techniques. Both techniques gave a 

large percentage of potentially identifiable marks across all substrates and ages (at 

the exception of the car cowling for which Wetwop outperformed VMD). Given that 

VMD requires extremely costly equipment and is a labor-intensive technique 

compared to powder suspensions, Wetwop is recommended to be applied when 

dealing with wetted non-porous surfaces. For those possessing a VMD machine, 

Wetwop could still be applied subsequently, in a sequence. 

Since several researches observed variations in the effectiveness of commercially 

available powder suspensions (in terms of ridge quality and background staining), 

Jones et al. compared four WPS formulas (i.e., TiO2 grade RG-15 - StanChem, Wet 

Powder white – Kjell Carlsson Innovation, Wetwop white – Armor Forensics, and 

Adhesive side Powder light - Sirchie) (244). The WPS formulas were applied on 

black insulating tape and the obtained results were analytically characterized using 

electron microscopy and X-ray photoelectron spectroscopy. The particle size 

distribution was quite similar for all brands (diameter range: 200-500 nm) and did not 

seem to play a role in the difference of effectiveness. On the other hand, the coating 

of the particles varies between the WPS formulations (in terms of morphology and 

chemical composition), and is more likely to explain the difference of performance 

observed between the different brands (245). In the same context, Jones et al. used 

atomic force microscopy and scanning electron microscopy to investigate the 

smoothness of surfaces classified as non-porous (i.e., formica, polyethylene, and 

unplasticised polyvinyl chloride) (248). They subsequently tried to correlate these 

analytical measures with the effectiveness of iron oxide powder suspension to detect 

fingermarks on those substrates (18-hour-old sebum-rich marks were used). They 

showed that both average roughness and topographical feature shape are important 

factors to consider for the processing of latent fingermarks. 

A detrimental effect of cyanoacrylate fuming has been observed if the items 

(latex/nitrile gloves or duct tapes) have to be subsequently processed by black 

powder suspension (233), as described in the “Cyanoacrylate fuming” section. 

256

2.2.6 Nanoparticles (nanopowders and nanocomposites) 

The use of nanoparticles certainly constitutes one of the major 

contributions in forensic science research these last years, with a sharp 

increase in number of scientific publications related to their integration in 

security documents, paints, inks, and most importantly new reagents to 

detect latent fingermarks (197, 250, 251). Numerous publications were 

consequently dedicated to the development of new methods based on 

gold nanoparticles (252-255), silica nanoparticles (256-259) (260), 

quantum dots (261-267), aluminium oxide (268), zinc oxide (269), 

phosphorescent compounds (270), titanium dioxide (244), and 

nanoparticles functionalized with antibody/antigen complexes (271, 272). 

The use of metal nanoparticles (e.g., gold and quantum dots) to detect 

latent fingermarks has been reviewed by Choi et al. (273). Nevertheless, 

most of the proposed techniques are still under development and have not 

yet succeeded in replacing classical reagents. It is certainly only a 

question of time before some of these new efficient nanoparticle-based 

detection techniques are integrated in casework. An overview of different 

kinds of nanoparticles is also presented in (274). 

Gold nanoparticles 

Works on gold nanoparticles were mainly driven by a readiness to enhance 

multimetal deposition (MMD), a method based on the use of colloidal gold to detect 

fingermarks on a wide range of substrates (275). First, the single-metal deposition 

technique (SMD), based on a simplified MMD process, has been optimized and is 

strongly recommended as a replacement for MMD (252). A luminescent version of 

the MMD has also been proposed, into which the silver coating has been replaced by 

a zinc oxide deposition step (253). This luminescent alternative is however still under 

development and cannot readily be applied in casework. Gao et al. proposed a one 

step MMD-like process to detect fingermarks, using glucose-capped gold 

nanoparticles and operating in a wider range of pH (254). On the contrary to what the 

authors claim, the mechanism looks more like a “gold-based SPR”, especially when it 

is said that it is working with blueish colloidal solution (this colour being a 

consequence of nanoparticles aggregation). Other authors trapped glutamatecapped 

gold nanoparticles in a lipophilic and cationic polymer (i.e., chitosan) to 

detect fingermarks after several hours of immersion (255). In this case, the way the 

fingermarks look like probably results from oil-enriched fingermarks. Extreme caution 

should be taken regarding the results. 

Quantum dots 

Quantum dots (QDs) are extremely luminescent nanoparticles, of one to 10 nm of 

diameter, which can be functionalized and solubilized in either aqueous solution or 

organic solvents. It is possible to identify three ways of integrating QDs in the 

detection of latent secretions: (1) as a dry powder, (2) as an aqueous solution, (3) 

embedded in a polymer that covalently binds to the secretion. Cadmium sulfide 

(CdS) encapsulated in a biopolymeric chitosan matrix has been used as a dusting 

powder for detecting fresh marks on aluminium foil (261). Luminescent fingermarks 

were obtained, but the use of dried cadmium-based nanoparticles as a dusting 

powder implies serious health and safety issues. Water soluble cadmium selenide 

257

(CdSe) QDs were synthesized and used to detect fresh marks on the adhesive 

surface of tape and promising results were obtained (262). Cadmium telluride (CdTe) 

QDs, synthesised in aqueous solution, were used to detect blood fingermarks on 

various nonporous surfaces such as aluminium foil, black polyethylene, glass or 

transparent polypropylene (265). A comparison with acid yellow 7, one of the best 

blood reagents for non-porous substrates, showed that the QDs were superior to acid 

yellow 7 on aluminium and equally effective for the other substrates. The immersion 

of a sample in an aqueous CdTe solution has also been proposed to detect 

fingermarks on non-porous substrates (263, 264). Several hours of immersion were 

however necessary, which constitutes too long a time. The embedment of QDs in a 

polyamidoamine dendrimer (PAMAM) has been successfully applied to enhance the 

contrast of fumed fingermarks on non-porous surfaces (266, 267). Nevertheless, an 

immersion time of several hours is required which is still much too long. As can be 

seen, the use of QDs to detect fingermarks currently remains a research topic and no 

casework uses of QDs have been reported yet. Some issues remain to be addressed 

before routine application, like the toxicity of cadmium and improved targeting of 

secretion residues by functionalization of the QD outer surface. 

Silica nanoparticles 

Silica nanoparticles constitute another kind of promising nanocomposites to detect 

fingermarks since a great freedom is offered in terms of outer functionalization and 

dye-doping. The use of luminescent silica nanoparticles for forensic purposes 

somewhat still remains at the pilot study stage. Theaker et al. recently reported 

entrapment of a variety of coloured and fluorescent dyes including basic red 28, 

basic yellow 40, fluorescein, methylene blue, oxazine perchlorate, rhodamine B, 

rhodamine 6G and thiazole orange within silica particles (256). The resulting doped 

nanoparticles were used in aqueous suspensions to detect fingermarks. The process 

is very similar to that described for small particle reagents. Micron-size particles also 

were used as dusting agents. Both fresh (20-minute-old) and aged fingermarks (40day-old) 

showed good definition after development. Liu et al. entrapped an europiumbased 

dye inside silica nanoparticles and further powdered the obtained 

nanocomposites to detect fresh and six-day-old latent fingermarks on various 

substrates (e.g., plastic bag, rubber glove, coloured paper, and green leaf) (257). 

Silica nanoparticles doped with carbon black have also been used as a fingerprint 

powder to detect fingermarks on glass and metal surfaces, before being lifted using a 

lifting tape (258-260). The originality of this work lies in the fact that the dusted marks 

were further imaged using surface assisted laser desorption / ionisation - time of 

flight - mass spectrometry (SALDI-TOF-MS). By doing this, it has been possible to 

differentiate fingermarks left by smokers (or drug users) from fingermarks left by nonsmokers, 

the silica nanoparticles performing as effective ionisation / desorption 

agents for the analysis of the drugs and metabolites contained in the secretion 

residue. 

Other nanoparticles 

Sodhi & Kaur chose to coat aluminium oxide nanoparticles with two different 

molecules: Lucifer Y (a fluorescent dye) and a natural hydrophobic substance (268). 

Their aim was to obtain a “nanopowder” to be dusted on substrates and 

characterized by an enhanced ability to detect fingermarks through lipophilic 

258

interactions. According to the authors, this nanopowder detects fingermarks on a 

wide range of surfaces (i.e., porous and non-porous, as well as white and 

multicoloured). It is particularly suitable for detecting fingermarks on glossy items, or 

on moist and sticky surfaces. 

Choi et al considered the use of ZnO nano-sized particles to be dusted or applied as 

small particle reagents (SPR) (269). Processed fingermarks were characterized by a 

visible fluorescence when illuminated with a long-range UV light source. ZnO-based 

SPR gave good results for all the tested surfaces (i.e., glass, polyethylene, 

aluminium), while dry dusting led to some background staining on polyethylene 

surface. When compared with conventional commercial powders, ZnO particles were 

less luminescent but showed excellent ridge detail, with minimum background 

staining. The authors also tried to dope the powder using lithium ions, to enhance the 

visible luminescence, but this did not significantly improve the results. 

Cheng et al. considered the use of two phosphorescent, surface-functionalized 

nanoparticles (i.e., doped LaF3 and ZnS) to detect trace biomaterials, such as the 

ones contained in fingermarks (270). The functionalization was such that the 

nanoparticles were water-soluble and functionalized with carboxylic groups (i.e. – 

COOH) to target amine groups from the latent secretions. Positive results were 

obtained for fresh fingermarks left on four tested substrates (i.e., aluminium, plastic, 

glass, and quartz). The detection procedure consisted in putting the substrates in 

contact with the nanoparticle solution for two hours. 

Finally, a new trend consists in functionalizing nanoparticles with biomolecules to 

take benefit of the highly selective antibody / antigen recognition process within the 

fingermark detection mechanism (271, 272). This strategy is further detailed in the 

next point. 

As described in the previous section, Jones et al. compared four brands of nanosized 

titanium dioxide to be used as powder suspensions on black insulating tapes 

(244). 

2.2.7 Antibody / Antigen recognition 

The use of antibodies constitutes a new and efficient approach to 

specifically target the secretion residue. We can distinguish methods using 

a direct antibody / antigen labelling procedure to target naturally secreted 

proteins (276, 277), from those using biofunctionalized nanoparticles to 

target drug metabolites present in the secretion (271, 272, 278). 

Hazarika et al. functionalized magnetic nanoparticles with a variety of antibodies to 

detect a range of drugs (i.e., THC from marijuana and methadone) or drug 

metabolites (of methadone and cocaine) in fingermarks left on glass slides (271, 272, 

278). The procedure requires the substrate to be in contact with the nanoparticles for 

15 to 30 minutes. One advantage of their method is that a magnet can be used to 

remove the excess of (magnetic) nanoparticles (the unbound ones). The remaining 

nanoparticles were fluorescently tagged before observation. Positive results were 

obtained for the four drugs and metabolites tested, with visible third-level details such 

as pores. 

259

Reinholz chose to target albumin, a protein constituting ca. 60% of the serum-protein 

volume and secreted by the eccrine sweat glands (277). More precisely, given that 

an “anti-human whole serum” was used, the whole serum constituents were actually 

targeted, and not specifically the albumin. Fingermarks were left on five porous 

substrates (i.e., white and manila envelopes, recycled paper, post-it, and 

nitrocellulose), and aged from 15 to 130 days under artificial and natural light. Once 

processed, purple fingermark ridges could be observed on a more or less clearer 

background (depending on the substrate), with visible third-level details. It should be 

noted that the detected marks progressively fade with time. Finally, the integration of 

this technique in the existing detection sequences was assessed. A DFO-processed 

fingermark (fresh mark on white paper) gave more details once subsequently 

processed with the albumin recognition technique. However, when applied after 

ninhydrin, the visible mark vanished and no detail could be further observed. Finally, 

this procedure gave equivalent results when applied after physical developer. The 

described procedure seems promising. Nevertheless, it is extremely time-consuming 

and labour-intensive, with several blocking/staining and rinsing baths. 

Drapel et al. identified and targeted three proteins present in the secretion residue 

and implicated in the skin regeneration process (i.e., keratin, cathepsin, and 

dermcidin) (276). Using a chromophoric enhancement process, positive results were 

obtained on fingermarks left on PVDF (a substrate commonly used in biology to fix 

proteins), but also on paper. Further research is currently performed, especially to 

decrease the time required by the procedure (several hours) and to introduce 

luminescent markers. 

2.2.8 Detection of fingermarks on thermal paper 

Some techniques specifically developed for thermal papers were 

described or renewed: incorporation of polyvinylpyrrolidones in a ninhydrin 

solution to avoid the darkening of the substrate (279), dry-application of 

1,2-indanedione (280), use of heat and steam (281, 282), iodine fuming 

(283), or vacuum metal deposition (282). The use of pDMAC to detect 

marks on thermal papers was also assessed (227), with the conclusion 

that classical amino acid reagents are better choices. 

In the previous report (284), we mentioned the “solution G3” as a powerful posttreatment 

for ninhydrin- and DFO-processed thermal papers whose reactive layer 

could have darkened due to the polar solvents contained in the reagent solutions 

(285). Recently, Schwarz & Klenke proposed a modified ninhydrin formulation 

containing polyvinylpyrrolidones (PVP) (279). The introduction of PVP in the working 

solution prevents the darkening of the thermal layer, and allows a one-step detection 

of fingermarks which readily appear in clear contrast with the background. PVP being 

nontoxic and reasonably priced, it could constitute a promising alternative to the 

existing two-steps discolouration procedure. One drawback of the currently proposed 

formulation is the ratio of polar solvent (compared to non-polar ones) that could 

cause ink running in particular cases. Further research is still undergoing, especially 

about the possibility to include PVP in DFO or 1,2-indanedione working solutions. 

260

The application of an amino acid reagent by “dry contact” with the thermal paper has 

been known for years. This technique simply consists in placing the substrate to be 

processed between two impregnated treatment papers, with no heat treatment (to 

avoid the substrate darkening). Recently, the use of 1,2-indanedione/ZnCl2 (Ind/Zn) 

has been proven compatible with this application protocol (280). Natural, nonenriched, 

fingermarks were left by one to four donors on unprinted and printed 

thermal papers (from supermarkets), and were processed between one and seven 

days after deposition. The use of an acetic acid-free Ind/Zn solution, with HFE-7100 

as the solvent carrier, gave the best results when the samples were left for 48 hours 

in contact with the treatment papers. It should be noted that HFE-7100 can be 

replaced by petroleum ether, but the quality of the developed fingermarks could 

decrease. 

The use of heat to detect fingermarks on thermal paper is one of the oldest methods. 

This technique is still used, as described by Scott who used a portable hair dryer 

combined with the application of steam during the heating process (281). 

Following their study about pDMAC, Lee et al. assessed the possibility of using this 

reagent to detect fingermarks on thermal papers (227). After having processed 

realistic depleted series of fingermarks on three kinds of thermal papers, they 

concluded that the best procedure to detect fingermarks on thermal paper consists in 

pre-dipping the sample into ethanol (warning: any text present on the substrate will 

be definitely lost) before using a classical amino acid reagent, such as DFO. 

Iodine is known to temporarily stain fresh latent fingermarks on a wide range of 

substrates. Recently, Jasuja & Singh announced that the use of iodine on thermal 

papers could lead to a permanent dyeing of the marks (brown / green colour), with 

marks up to one-year-old successfully detected (283). Their hypothesis was that 

iodine molecules trapped in the latent sebaceous secretions could promote the 

oxidation of the leuco-dyes molecules contained in the thermal paper layers. 

According to the authors, sebaceous marks react more intensely than eccrine ones. 

Vacuum metal deposition (VMD) or steam – by placing the sample in an incubator for 

ca. five minutes – were used to detect sebum-rich latent fingermarks, up to threeweek-old, 

on thermal paper (282). Both techniques offer the advantage of not 

causing a darkening of the thermal paper, as it can be observed with classical 

chemical methods containing polar solvents. 

2.2.9 Detection of fingermarks on metal and cartridge cases 

In the context of fingermark detection on metal (and therefore, cartridges), 

John W. Bond is certainly the major contributor of these last three years, 

with several publications describing the metal corrosion effect caused by 

latent secretions (286-293). Besides these works, a statistical study 

showed that the rate of recovery of fingermarks on firearms and cartridges 

(using cyanoacrylate and dye) was actually lower than expected (294), 

and another study aimed at determining the best sequence to be applied 

on brass and nickel cartridge casings (295), without success. 

261

The metal corrosion effect 

The metal corrosion effect is based on the discolouration of metal surface through 

the action of an oxydizing element, more particularly the ionic salts contained in 

latent secretions. When a metal surface bearing latent fingermarks is heated up to 

400-600°C, the marks become visible as a consequence of the metal corrosion 

induced by sodium chloride, in particular (287). The corrosion can also take place in 

air at room temperature, several days after fingermarks were deposited (287). 

Ambient humidity and oxygen are required for the corrosion to occur (292), as well as 

heat to obtain good ridge resolution. The success of the technique described by 

Bond et al. relies more on the composition of the metal (especially in terms of 

thermodynamic consideration, such as the redox potential) and on the quantity of 

salts in the secretion, rather than on the age of the fingermarks. Moreover, an 

effective (and therefore destructive) removal of the fingermark deposits from the 

surface is required before the corroded region could be processed. To this purpose, it 

was determined that immersing the sample in warm soapy water while rubbing with a 

tissue constituted the best technique so far to remove all traces on brass without 

affecting the properties of the metal surface (293). Once corroded, the marks are 

durable and resist any further cleaning attempt. This technique seems particularly 

suitable for samples found in arson scene or contaminated by paint (e.g., spray 

painting above fingermarks) (287). 

Preliminary tests were performed on brass, copper, steel and aluminium disks using 

non-enriched fingermarks, aged from less than five minutes to seven days, left by 40 

donors (287). For copper, temperatures should not exceed 400°C to avoid oxide 

flaking. On steel, fingermarks were of blue/dark appearance, certainly due to a 

chemical reaction with the metal (passivation or blueing). On aluminium, no 

fingermark could be detected, certainly due to the easiness of surface autopassivation 

preventing the corrosion to take place. In a second time, ten metal 

surfaces (i.e., aluminium, copper, gold, lead, magnesium, nickel, silver, tin, titanium, 

and zinc) and ten metal alloys were processed and compared (286). No heat was 

used, but the fingermarks (left by 40 donors) were aged at room temperature for ten 

days before being washed using water and a detergent. Spontaneous corrosion led 

to the observation of well-resolved fingermarks on some substrates (e.g., Ni, Sn, Pb, 

Cu, Ag and Au). The presence of chloride ions in eccrine sweat led to the formation 

of metal halide complexes, which enhanced the degree of noble metal corrosion 

(e.g., gold or silver). Finally, metal alloys containing more than 40% copper gave 

satisfactory results, with a good contrast level between the corroded and noncorroded 

parts, and the presence of third-level details (286). 

An enhancement procedure, based on electrostatic attraction, was also described 

(287-289, 291). This technique relies on: (1) the possibility to apply an electric 

potential (i.e., 2.5 kV) to metal objects, and (2) the preferential adherence of 

conductive carbon powder (10 µm, adsorbed on spherical beads) to the corroded 

parts of a metal surface (the corrosion occurring naturally with time due to the 

presence of fingermarks on the surface, as described above). This technique 

nevertheless requires washing the samples before they are processed, using water 

and acetone, to remove the remaining fingermark residue (and only leave the 

corroded part). The detection of fingermarks was successful for brass and copper 

disks, but failed for steel and aluminium. It should be noted that the authors 

compared this technique with small particle reagent (SPR), but after having cleaned 

262

all the samples. SPR logically failed in detecting fingermarks due to this destructive 

treatment. The comparison would have been more meaningful by applying SPR 

without having thoroughly cleaned the samples first. 

When applied on fired brass cartridge cases, the metal-corrosion technique gave 

some positive results, but almost none, certainly due to the physical abrasion and 

increased temperature that occurred during the shot (287). The author also published 

a casework performed on four 14-year-old discharged brass shell casing found at a 

crime scene (290). During the investigation, cyanoacrylate fuming combined with dye 

staining have been applied without success. In this casework, the author decided to 

heat the casings up to 700°C to remove the cyanoacrylate deposit and to induce the 

corrosive reaction between the brass surface and any fingermark that could be 

present on it. The application of the electrostatic enhancement (2.5 kV), described 

above, led to the observation of some ridges. 

Other techniques 

Edmiston & Johnson investigated the best sequence to be applied for detecting latent 

fingermarks on brass and nickel cartridge casings (295). The study has been 

performed on fired cartridges that were first cleaned, before fresh fingermarks were 

left by four donors and immediately processed (ten casings per tested sequence). It 

was concluded that the best-so-far sequence to be applied on brass cartridges was 

“Cyanoacrylate - Powder - Acidified Hydrogen Peroxide - Rhodamine 6G”, and for 

nickel cartridges “Cyanoacrylate - Rhodamine 6G - Acidified Hydrogen Peroxide - 

Powder”. During a “realistic test”, cartridges manipulated by bare hands were shot, 

and processed according to the two sequences described above. As a result, no 

identifiable marks were detected on the brass casings, and two potentially identifiable 

marks were detected on the nickel casings. The authors concluded that the 

sequence proposed for brass casings may not be optimal, given the experimental 

differences between the first (fresh fingermarks deposited on clean fired casings) and 

second part (older fingermarks deposited before the shot) of the study. Further 

studies are thus required. 

A statistical study showed that the rate of useable fingermarks detected on firearms, 

cartridges, or magazines processed by cyanoacrylate fuming followed by dye 

staining was quite low (294). The data provided by the Minneapolis Police 

Department Crime Laboratory covered a period of one year (from September 2006 to 

September 2007), representing a total of 3,436 items. Twelve percent of all firearms 

and twelve percent of all magazines led to useable fingermarks (35 and 19 marks, 

respectively). No fingermarks were useable on unfired cartridges, and only one mark 

was useable on a discharged cartridge casing (however, the mark is believed to have 

been left after the shot). 

2.2.10 Detection of fingermarks on tapes/adhesives 

The use of Un-du followed by the application of Wetwop was determined 

as being the best-so-far sequence to separate stuck adhesives and to 

subsequently detect latent marks (296). As an alternative to Un-du, 

submersing stuck adhesives in liquid nitrogen for 30 seconds could help in 

separating them (297). It was also observed that freezing adhesives 

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efore powdering them (while still cold) could help in detecting latent 

fingermarks (298). Finally, a comparative study was performed between 

four commercially available white powder suspensions (244). 

Molina tested the effectiveness of Un-du to separate stuck adhesives, and its effect 

on the subsequent detection techniques (i.e., gentian violet, Sticky-side Powder, and 

Wetwop) (296). Fingermarks were left on eight brands of adhesives and one selfadhesive 

stamp, which were further stuck on white paper and ironed for 2-3 minutes 

to induce immediate adhesion. Un-du was subsequently applied before the samples 

were processed. The results showed that excessive amount of Un-du could cause 

detrimental effects on the adhesive backings. All attempts provided latent 

development, but the best results were obtained by combining Un-du with Wetwop, if 

the latter was rinsed immediately after its application to avoid severe and irreversible 

background staining (this procedure could be repeated if required). 

The use of cold to help in detecting fingermarks on adhesives has been studied by 

Cramer & Glass (298). The study consisted in freezing (5.5°C) four types of 

adhesives (i.e., duct, clear packing, scotch and electrical tapes) bearing fingermarks 

for one to 120 minutes before being dusted immediately with black or magnetic 

powder. Successful detection was obtained in each case. Nevertheless, it seems that 

the operator was not experienced with fingermark powdering (according to the 

authors), leading to difficulty in evaluating the results at the beginning of the 

experiment, and no comparisons were made between frozen and non-frozen 

samples to really assess the role played by the freezing of the samples. 

Bergeron conducted a study about the use of liquid nitrogen to separate adhesive 

tapes that have stuck together (297). Considering 20 different brands of tape, the 

author succeeded in separating stuck adhesives after having submersed the tapes 

for 30 seconds in liquid nitrogen, followed by the detection of latent fingermarks using 

Sticky-Side Powder once the adhesives were brought back to room temperature. It 

should be noted that some difficulties could be encountered with some adhesives 

(e.g., no separation or breaking of the adhesive into pieces). The author also studied 

the effect of environmental conditions (e.g., heat, rain, humidity) with samples aged 

up to 36 days. When the samples were protected from extreme conditions (e.g., kept 

in laboratory), successful results could be obtained, even after 36 days. 

Nevertheless, when subjected to outdoor environments, the likelihood of finding 

latent marks was poor after a few days. Another interesting observation was made: a 

mirror image of a mark could be detected on the opposite adhesive side (i.e., the one 

that was stuck on the fingermark). This should be kept in mind. 

As described in a previous section, Jones et al. reported an analytical study in which 

they compared different white powder suspensions (i.e., TiO2 grade RG-15 - 

StanChem, Wet Powder white – Kjell Carlsson Innovation, Wetwop white – Armor 

Forensics, and Adhesive side Powder light - Sirchie) used to detect fingermarks on 

black insulating tape, and tried to explain the differences of quality in terms of 

chemical coating (244). 

264

2.2.11 Detection of fingermarks on items collected from arson scenes 

Several authors assessed the possibility to detect fingermarks in arson 

scenes (299 pp. 12-31, 2009] #483). The conclusions were quite 

encouraging since many concluded with the fact that fingermarks are quite 

resistant to elevated temperatures and could be retrieved if the adequate 

procedure is followed (i.e., soot removal process and detection 

techniques). Physical developer and black powder suspension were 

identified as being the most suitable techniques to be applied on porous 

and non-porous substrates, respectively (299 pp. 12-31, 2009] #483). On 

paper, DFO was recommended for exposure temperatures below 100°C, 

and physical developer above 100 °C (300). 

With respect to marks in blood, protein stains were shown to be still 

efficient up to 200 °C, on the contrary to heme reagents (such as LCV) 

(301 pp. 64-82, 2009] #577). Acid black 1 and acid violet 17 were 

recommended for porous and non-porous substrates, respectively. Tests 

performed with bloodstains in an apartment-like structure showed that 

luminol, fluorescein, and Bluestar were still efficient in less burned areas 

(302), but failed to detect blood marks in the most burned area. 

Some authors observed that fingermarks on paper could become naturally 

fluorescent once exposed to 160-180 °C (300, 303). Below or above these 

temperature ranges, this self-fluorescence could no longer be observed. 

Silicone casting and Absorene (a commercial product) were both 

recommended as soot removal techniques (299 pp. 12-31, 2009] #483, 

301 pp. 64-82, 2009] #577), as well as air-sprayed liquid latex (304, 305). 

Bradshaw et al. conducted a study about the recovery of fingermarks from arson 

scenes, and more particularly: the range of temperatures and exposure times at 

which latent fingermarks could survive, the soot removal process, and the efficiency 

of the fingermark detection techniques (299 pp. 12-31, 2009] #483). Various 

substrates, representative of what can be found in household equipment, were used 

(e.g., paper, ceramic, metal, smooth melamine). On each substrate, several donors 

were asked to deposit depletion series of “natural” fingermarks (the marks were not 

artificially enriched with sebum by rubbing the fingers on the face, for example). 

Eighteen hours after deposition, the fingermarks were exposed at 100 to 300 °C in a 

laboratory oven (for one to seven hours), at 400 to 600 °C in a high-temperature 

furnace (same time interval), or directly put in a simulated fire that was extinguished 

with water (the temperature reached up to 900 °C). The authors demonstrated that 

fingermarks can still be detected after a prolonged exposure at 200°C using most of 

the current detection techniques (i.e., DFO, ninhydrin, physical developer - PD, dry 

powders and powder suspensions, cyanoacrylate, and vacuum metal deposition - 

VMD). Above this temperature, the detection of latent fingermarks is still possible, but 

the range of techniques that can be applied is limited to cyanoacrylate (if the 

substrate has not been wetted) and VMD. In most of the cases, the most suitable 

techniques to be applied on porous and non-porous surfaces were physical 

developer and black powder suspension, respectively. The authors do not 

recommend to re-humidify the fingermarks (e.g., by breathing on it) before applying 

powders, since it was found to have a detrimental effect on several marks. If soot has 

to be removed prior to the processing of the surface, the authors recommend lifting 

tapes, silicone casting and Absorene (a commercial product used for cleaning 

paper). Washing with 4% sodium hydroxide gave good results but should be used in 

265

combination with a mechanical technique, such as lifting tape. Washing under water, 

ultrasonic bath, or use of organic solvents (e.g., chloroform or xylene) are not 

recommended since they induce damage to latent fingermarks. It should finally be 

noted that the soot can make fingermarks already visible. A careful examination 

should therefore precede any further treatment (soot removal or application of a 

fingermark detection technique). 

The possibility to recover fingermarks on paper exposed to elevated temperatures 

was also conducted (300). Five donors deposited non-enriched fingermarks on white 

paper. The marks were allowed to age from one hour to one month before being 

heated to temperatures ranging from 50 °C to 200 °C, for 10 to 320 minutes. 200 °C 

has been chosen as maximum temperature since paper auto-ignites at ca. 233 °C. 

DFO, ninhydrin and physical developer were further applied when attempting to 

detect latent fingermarks. The results showed that some fingermarks were readily 

visible after the heating process, especially at 150°C and 200°C. Moreover, the 

marks subjected to 150 °C were naturally fluorescent when excited at 473 to 548 nm 

and observed using a 549 nm filter (no fluorescence was however observed for the 

substrates exposed to 50, 100, and 200 °C). Successful results were obtained with 

the chemical methods for exposure conditions up to 200 °C for ca. five or six hours. 

DFO is the recommended technique for exposure temperatures below 100°C, and 

physical developer above 100 °C. The substrate could also be processed with PD 

subsequently to DFO. If the substrates have been wetted during the extinguishing of 

the fire, PD is to be applied whatever the exposure temperature. 

The natural fluorescence of fingermarks on paper exposed to elevated temperature 

has been further studied by the same team (303). Three kinds of fingermarks (i.e., 

eccrine, sebaceous, and “natural”/ungroomed) on white and filter papers were 

exposed to temperatures ranging from 110 °C to 190 °C, for 20 minutes. It was found 

that eccrine sweat was most subjected to fluorescence between 130 to 180 °C, with 

a maximum intensity obtained for the 160-180 °C range. It is hypothesized that 

thermal degradation of some amino acids could be at the origin of the observed 

luminescence. Sodium chloride and urea also fluoresced upon exposure to elevated 

temperature. Optimal observation conditions were determined as being: excitation 

source of 352 to 509 nm, combined with an observation filter of 510 nm. 

Considering the effect of fire on fingermarks in blood, Moore et al. showed that some 

of the commonly used protein stains (i.e., acid black 1, acid yellow 7, and acid violet 

17) continue to be efficient in detecting marks in blood for exposure of the 

fingermarks up to 200°C, while heme-specific reagents (such as LCV) stopped giving 

positive results between 100 and 150°C (301 pp. 64-82, 2009] #577). Various 

substrates were considered (e.g., paper, cotton fabric, ceramic tile, smooth 

melamine, metal) and defibrinated horse blood was used to create depletion series of 

blood fingermarks. Eighteen hours after deposition, the fingermarks were exposed to 

100 to 300 °C in a laboratory oven, and to 400 to 600 °C in a high-temperature 

furnace, for one to eight hours. Commonly used detection techniques were tested, 

specific for blood or not (i.e., acid black 1, acid violet 17, acid yellow 7, DFO, 

Leucocrystal violet - LCV, PD, VMD, cyanoacrylate with dye, solvent black 3, and 

powder suspensions). The recommended techniques are acid black 1 and acid violet 

17 for the processing of porous and non-porous surfaces, respectively. Interestingly, 

above 200°C, it is still possible to detect some marks but it becomes impossible to 

266

know if the mark was originally made of blood or not (due to a baking effect). Vacuum 

metal deposition has been shown to detect fingermarks up to 900°C on the ceramic 

tiles. Cotton fabric proved to be a very difficult surface, with none of the techniques 

giving ridge details. On this substrate, only acid yellow 7 gave luminescent spots 

above 150°C, even if it is commonly not recommended to be used on porous 

surfaces. In terms of soot removal techniques, their conclusions were similar to the 

above-described study, with silicone casting and Absorene. Contrary to classical 

latent marks, the use of lifting tape is not recommended since blood deposits could 

be lifted off. 

The effect of fire exposure on bloodstains (and the subsequent chemical 

enhancement and DNA profiling) has been studied by Tontarksi et al. (302). An 

apartment-like structure was built, various bloodstains and marks were placed on 

walls or furniture, before arson was ignited (i.e., sofa fire) with burning developed for 

ca. 45 minutes, to be eventually extinguished using water. Once the place cooled, 

changes in the previously applied bloodstain patterns were documented, chemical 

enhancement techniques were used (i.e., luminol, fluorescein, and Bluestar), and 

samples were collected for DNA analysis. During the fire, temperatures from 146 °C 

up to 923 °C were recorded in different areas of the apartment. In the most burned 

area, the chemical reagents gave negative results and no bloodstains could be 

observed using white light. Nevertheless, in less burned areas, bloodstain patterns 

were observed using a bright light and positive results were obtained with all the 

chemical enhancement techniques. Nevertheless, fluorescein is recommended to be 

applied first, since it requires less reagent to give a positive observable result 

(compared to luminol and Bluestar). Hemastix tests were conducted to assess the 

presence of blood, and gave positive results on stains visible with a bright light. It 

was sometimes necessary to remove the soot to obtain a positive result (done by 

wiping with 70% isopropanol or distilled water). As to DNA profiling, the results were 

primarily positive, except for some samples exposed to high temperatures and/or 

close to the ignition source (for measured temperatures above 845 °C, no DNA 

profile was obtained). The authors also showed that none of the chemical 

enhancement reagents interfered with the ability to obtain a DNA profile. The authors 

recommended trying to obtain DNA profiles even for stains that yield negative results 

after chemical screening tests for blood. This is especially true for the samples close 

to the fuel source that yield DNA profiles suitable for comparison despite negative 

indicative tests. 

Liquid latex was air-sprayed on an arson scene to remove the excess of soot 

resulting from the fire, before processing the surfaces to detect latent blood marks 

supposedly lying beneath (304). The procedure consisted in spraying a thin layer of 

latex, letting it dry for five to ten minutes, peeling it off once dry, and examining the 

surfaces for latent marks using an alternative light source. On painted doors or 

glossy surfaces, 90% of soot was removed by following this method, allowing the 

observation of the underlying fingermarks or bloodstains. On untreated woods and 

wallpapers, the rate of removal dropped to 80 and 70%, respectively. In some cases, 

the soot acted like a fingerprint powder and remained on the fingermark after the 

peeling of the latex. This method could be considered as an inexpensive alternative 

to sodium hydroxide washings, which could be destructive on some marks (such as 

blood marks, for example). Nevertheless, further experiments on the effect of such 

treatment on the subsequent chemical detection methods have to be carried out. 

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Spraying of liquid latex was also used by Clutter et al. who assessed its efficiency in 

removing soot on glass and drywall board (wallboard) and its effect on a subsequent 

ninhydrin treatment to detect fingermarks (305). Latent marks and blood spatters 

were deposited on both substrates before being almost immediately placed near a 

controlled burn (ca. 500 °C, high level of soot). The items were then removed so that 

no water was poured on them. After the items cooled, latex was sprayed, let dry and 

peeled off. Ninhydrin was then sprayed on the wallboard, while the glass was 

powdered using black and magna powders. If soot made some fingermarks already 

visible before any soot removal attempt, ninhydrin and powders failed in detecting 

new marks after the application of latex to remove soot. It should be noted that 

ninhydrin was not tested on wallboard to confirm that this technique is suitable to 

detect fingermarks on this kind of substrate. 

2.2.12 Blood marks - Fingermarks in blood and detection of bloodstains 

Few studies were dedicated to the detection of marks in blood, certainly 

reflecting the quality of the currently available detection techniques. The 

effect of fire exposure on blood marks has however been studied by 

various authors (301 pp. 64-82, 2009] #577, 302, 306). An original study 

aimed at optimizing the deposition procedure of fingermarks in blood 

(307). Finally, a new method to lift blood marks from difficult substrates 

(like garments) has been proposed (308). 

The influence of the deposition procedure on the ridge quality of blood marks has 

been investigated (307). The conclusions of this study could be of a first interest 

when conducting a study on blood detection techniques. To perform this trial, the 

author took into account several parameters such as the deposition pressure, the 

quantity of blood on the finger, the drying time before the deposition, or the 

orientation of the surface. This study showed that more marks of evidential value 

could be obtained when small amounts of blood were loaded on the finger (i.e., 10 to 

20 µL), and allowed to dry before being left on a vertical surface (glossy white poster 

board was used in this study) with some increased pressure. It should be noted that 

air temperature, airflow, blood and skin temperatures also affected the drying times 

and the resulting ridge details. The author also observed that negative marks (white 

ridges and bloody inter-ridges) could be obtained when first removing the excess of 

blood on the finger, then allowing the blood on ridges to dry before pressing the 

finger on the surface (the blood trapped in the inter-ridge furrows remaining wet). It 

has also been shown that the mechanism of formation of negative marks is not 

related with the pressure that would force the blood off the friction ridges and into the 

furrows during deposition, as it is commonly thought. 

The effect of fire exposure on blood marks (and the subsequent chemical 

enhancement and DNA profiling techniques) has been studied by various authors 

(301 pp. 64-82, 2009] #577, 302, 306). These works are detailed in the “Detection of 

fingermarks on items collected from arson scenes” section. 

The strong chemical affinity of TiO2 for blood has been known for several years (it 

has especially been used to detect blood marks on non-porous substrates (309)). 

Recently, lifting strips containing titanium dioxide (TiO2) were successfully applied to 

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lift blood fingermarks from several porous or non-porous substrates (308). In this 

study, lifting strips were made by fixing different chemical products (conventionally 

used in blood detection techniques) to a nylon membrane: titanium dioxide, Kodak 

Photo-Flo, 5-sulfosalicylic acid, Liqui-Nox, polymers, and water. Bloody impressions 

were left on six substrates (i.e., three non-porous - aluminium, black plastic, and 

metal, two semi-porous - black leather and wood, and one porous - cotton), and were 

aged from one hour to six months before being lifted and processed. The lifting strips 

successfully lifted blood impressions from all six substrates up to one month after 

their deposition, with visible identifiable ridge details (except for the stained wood for 

which ridge details were observed for up to one-week-old marks only). An advantage 

of these lifting strips is that the lifted mark can be further processed using the 

conventional blood detection techniques. 

2.2.13 Fingermark detection and DNA or biological fluid analysis 

The interactions between fingermark detection techniques and DNA 

recovery/profiling are always of a first interest when both evidence types 

are to be retrieved. On this subject, it has previously been shown that 

most of the fingermark detection techniques have no detrimental effect on 

the DNA profiling (310-312), at the exception of shortwave UV light, DAB, 

or physical developer (12). Nevertheless, the quantity of DNA seriously 

drops after the application of fingermark detection techniques. Finally, it 

seems that no correlation exists between the ability of an individual to be a 

good fingermark donor (i.e., donation status) and the quantity of DNA that 

can be recovered from his fingermarks (i.e., shedder status) (313). 

Sewell et al. investigated the effect of ninhydrin and 1,8-diazafluoren-9-one (DFO) on 

the recovery of DNA profiles recovered from fingermarks on paper (311). For this 

study, fresh sebum-loaded fingermarks from two donors were deposited on filter and 

office papers, glossy magazines, white card and newspaper. Two commerciallyavailable 

DNA profiling techniques were compared: QIAmp DNA mini kit and DNeasy 

plant mini kit (both from QIAGEN). A significant increase in DNA recovery has been 

observed by using the DNeasy kit as compared to the QIAamp kit, for all treatments 

and all paper types. This study also showed that the largest amounts of DNA were 

recovered from the magazines, on the contrary to white and office papers that 

interfered with the recovery process. Almost no DNA was recovered from the office 

paper, and no profile could be obtained. Full profiles were however obtained for 

traces on filter paper, magazines and newspapers. These observations were similar 

to previous studies on this topic (310, 312). Finally, a decrease of 60% in DNA 

recovery was observed after the application of fingermark detection techniques, 

especially on the filter paper and magazines. Nonetheless, these treatments were not 

shown to interfere with the PCR amplification and DNA profiling steps. 

Dominick et al. investigated the possible relationship between the fingermark 

donation ability of an individual and his DNA shedder status (313). For this study, 

eight donors were asked to deposit ten series of seven successively deposited 

fingermarks (i.e., without recharging the fingertips with secretions) on glass. For 

DNA, they were asked to hold a tube for several seconds. Fingermarks were 

processed using aluminium powder and ranked according to their quality. DNA was 

swabbed from the tubes before being extracted and amplified. No relationship was 

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found between the fingermark donation status and the DNA shedder status. The level 

of ridge details on a fingermark is consequently not correlated with the subsequent 

DNA profiling tests. 

2.2.14 CBRNE-related evidence (Chemical, Biological, Radiological, Nuclear, and 

Explosive) 

Research on CBRNE-contaminated evidence has seriously increased 

during these last three years. Most of the researches were dedicated to 

the effect of the contaminant, and of the decontamination procedures, on 

the subsequent detection of latent fingermarks. In the case of chemical 

contamination, if items were contaminated with acid chemical vapours, the 

use of volatile organic bases could help in improving the cyanoacrylate 

fuming process (314). In the case of biological contamination, the use of 

formaldehyde vapours during the decontamination procedure could be 

detrimental for some amino-acid reagents (315). No adverse effects were 

however observed if irradiation is used to decontaminate the items (316). 

Finally, bacterial activity showed almost no detrimental effect on the latent 

and blood fingermarks left on porous and non-porous substrates 

(variations were however obtained for adhesives) (317). In the case of 

irradiation, fingermarks could still be detected on glass and aluminium 

after high doses of irradiation (even if glass could become optically 

degraded) (318). However, detrimental effects induced to the substrate 

itself (especially for plastic and paper items) could hamper the fingermark 

detection processing. Nevertheless, up to 100 kGy, it has been shown that 

the processing of latent fingermarks using common detection techniques 

is still possible. In another study, some chemical and physical 

decontamination procedures were shown to be highly detrimental on the 

subsequent recovery of latent fingermarks on documents (319). In such 

cases, it is either recommended not to decontaminate the documents 

(even after the recovery of fingermarks), or to proceed to the 

decontamination after the recovery of fingermarks. Explosive 

contaminations could be located and identified on latent fingermarks using 

analytical tools such as Raman chemical imaging (320), or Fourier 

transform infrared spectroscopy (321, 322). Fingermarks on post-blast 

devices could still be detected using either cyanoacrylate fuming (323), or 

reflected UV imaging (RUVIS) (324). 

Chemical contamination 

The exposure of evidence to hydrochloric acid (HCl) vapours or chlorine (from a 

chemical weapon or from methamphetamine laboratories) could result in an 

acidification of the secretion residue, causing detrimental effects on some fingermark 

detection mechanisms (such as cyanoacrylate polymerization or ninhydrin). 

McDonald et al. assessed the efficiency of a basic pretreatment (using volatile bases) 

prior to the application of detection techniques (314). Fingermarks on glass slides 

and standard photocopy paper were considered. They observed that exposure to 

high level of acid vapors could indeed lead to an inhibition of the cyanoacrylate 

polymerization and to a modification of the ninhydrin reaction mechanism (leading to 

light pink-red marks instead of dark purple). Exposure of the evidence to 

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triethylamine and ethanolamine vapors prior to cyanoacrylate fuming leads to much 

more effective fingermark detection. Nevertheless, an increase of the background 

staining has been observed, especially if the evidence has not been exposed to acid 

environment prior to the basic pretreatment. Physical enhancement techniques (e.g., 

powder dusting or small particle reagents) could successfully be applied on the 

acidified substrates without any pretreatment step, but there is a risk that increased 

background staining occurs. 

Biological contamination 

Hoile et al. proposed to modify a decontamination protocol based on the use of 

formaldehyde, which acts like a biocidal/sporicidal agent and which is generally used 

in the case of biological contaminations (e.g., anthrax spores) (315). When dealing 

with an item suspected to have been contaminated with biological agents, a common 

decontamination procedure consists in using formaldehyde vapour for six to 12 

hours. This treatment results in the destruction of the spores, but also to a 

degradation of the amino acids present in the latent residue. Hoile et al. studied the 

effect of this decontamination procedure on some commonly used fingermark 

detection techniques (i.e., ninhydrin, DFO, 1,2-indanedione, physical developer, 

cyanoacrylate, and powders). For this study, fingermarks were left on glass and 

paper substrates, which were put in contact with formaldehyde vapors from 10 

minutes to 12 hours. If cyanoacrylate fuming, powdering, and physical developer 

continued to be successfully applied after the decontamination procedure, this was 

not the case with ninhydrin, DFO, and 1,2-indanedione which failed to detect latent 

marks. Hoile et al. revised the decontamination procedure by reducing the 

concentration of formaldehyde and the exposure time (i.e., 40 minutes instead of six 

to 12 hours), which resulted in 66%, 33%, and 8% successful recovery rates for 

ninhydrin, DFO, and 1,2-indanedione, respectively. 

The impact of bacterial agents on the efficiency of the classical fingermark detection 

techniques has been studied by Wilkinson et al. (317). For this study, various porous, 

non-porous and adhesive substrates were considered (e.g., plastic, glass, cardboard, 

metal, paper, tapes) on which latent and blood fingermarks were left. A high number 

of fingermarks (ca. 400) from eight donors were used, and were aged for one to 

seven days prior being exposed to the bacterial agents. The samples were 

contaminated with bacterial cultures for 24 to 48 hours, before being processed with 

classical fingermark detection reagents (i.e., DFO, ninhydrin, amido black, Hungarian 

red, leucomalachite green, powder suspension, cyanoacrylate fuming, and dry 

powders). The authors observed no effect of the bacterial exposure for porous 

exhibits (100% trace recovery), and a minor effect on nonporous substrates (97% 

trace recovery for cyanoacrylate). Blood reagents reacted quite well, with a rate of 

recovery between 86 and 97%. Only the adhesive items showed varied results. In 

addition to these observations, the bacterial exposure time (from 24 hours to 48 

hours) and the age of the fingermarks (from one to seven days) seemed to have no 

effect on the recovery rates. 

Hoile et al. investigated the use of gamma irradiation (25 to 40 kGy using cobalt-60 

as irradiation source) to decontaminate spore-contaminated evidence, and its effect 

on the subsequent fingermark detection techniques, as well as on the recovery of 

human DNA (316). Different substrates were considered (i.e., paper, cardboard, 

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glass, and plastic) on which fingermarks from three donors were left. After having 

been exposed to irradiation doses from 200 Gy to 5 kGy, the items were processed 

using common detection techniques (i.e., ninhydrin, DFO, 1,2-indanedione, physical 

developer, cyanoacrylate and dry powders). Preliminary tests on microbial colonies 

showed that a gamma dose of more than 3 kGy should be used to decontaminate 

evidence. As observed by Colella et al. (318), glass underwent some discolouration 

(browning) above 1 kGy without interfering with the quality of the fingermark 

detection process. No adverse effect due to irradiation (in terms of ridge details) was 

observed for the processing of the porous and non-porous substrates, even for a 40 

kGy irradiation dose. This observation is in agreement with the results obtained by 

Colella et al.. About DNA profiling, the quantity of DNA recovered could be affected 

by the irradiation process, but the possibility to recover human DNA from paper, post 

gamma irradiation, is not excluded even up to 50 kGy doses (more severe 

degradation occurs above this value). 

Radiation 

Colella et al. investigated the possibility to recover latent fingermarks from evidence 

that was exposed to ionizing radiation (318). Common surfaces were considered 

(i.e., aluminium, glass, office paper, and plastic), on which fresh and good 

fingermarks were left, before being irradiated to doses from 1 to 1000 kGy (which 

constitutes significantly higher exposure doses compared to the electron beam 

irradiation used to sanitize mail). A cobalt-60 ionization source was placed at one, 10 

and 100 mm of the substrates, and exposure times ranged from 24 hours to seven 

days. Common detection techniques were considered (i.e., cyanoacrylate with dye, 

black powder, ninhydrin, DFO, 1,2-indanedione, and physical developer). It was 

concluded that radiolysis had a considerable effect on the quality of the developed 

fingermarks, at the exception of marks on glass and aluminium which showed no 

influence due to the ionization process. With respect to glass, the irradiation caused 

an optical degradation of the substrate which became deep brown as the ionizing 

dose increased. Nevertheless, this effect had no impact on the detection of 

fingermarks (the only detrimental effect would be in terms of reduced contrast). For 

the other substrates, the irradiation caused physiochemical and mechanical damage 

in the inner structure of the material (e.g., plastics) especially for doses above 100 

kGy. This resulted in unwanted polymerization sites or background fluorescence, in a 

degradation of the substrate mechanical properties preventing them to be properly 

processed (e.g., paper), or in a chemical modification of the secretion residue making 

them less prone to react. Nevertheless, up to 100 kGy, it has been shown that the 

processing of latent fingermarks using common detection techniques is still possible. 

Parkinson et al. investigated the efficiency of two decontamination procedures used 

in case of irradiated documents, and their impact on fingermark detection techniques 

and ink analysis (319). Three irradiation sources were used (i.e., cesium-137, 

americium-241, and strontium-90) to contaminate paper samples that were 

subsequently processed for fingermark detection (using DFO, ninhydrin, 1,2indanedione, 

and physical developer). After the detection of fingermarks, the 

substrates were decontaminated by following a physical process (scraping of the 

surface with a scalpel followed by the use of a pencil eraser) or a chemical one 

(immersion of the evidence from 0 to 30 minutes in a sonication bath containing 

water or cyclohexane, and DEZ-1 - a complexing agent used as a decontaminant). 

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The use of DEZ-1 allowed a decontamination rate of 97% for both solvents (water or 

cyclohexane), but total decontamination could not be reached since a radiation level 

of 10 times the background level always remained (even at higher concentration of 

DEZ-1, and for longer sonication times). The optimized concentration of DEZ-1 has 

been evaluated at 2 % for an immersion time of five minutes. Increasing these 

parameters does not improve the decontamination rate but induces a degradation of 

the paper structure. The physical method was unsuccessful in decontaminating the 

cesium-137 and strontium-85 isotopes, with only 1.2% and 0.2% decontamination 

rates, respectively. However, it proved to be highly successful in removing the solid 

contaminating isotopes (yellowcakes), with 99% decontamination rates compared to 

the contaminated fingertips on an aluminium-coated microscope slide. The imaging 

was performed by comparing the spectra obtained from the scanned fingermarks 

with the reference spectra of the targeted explosives, using Pearson's crosscorrelation. 

In this preliminary study, it was possible to locate and identify explosives 

from contaminated fingermarks. The consideration of realistic surfaces and the effect 

of fingermark detection techniques (such as cyanoacrylate) on the subsequent 

identification of explosives have to be investigated in further experiments. 

Attenuated total reflection – Fourier transform infrared (ATR-FTIR) was used to 

locate and identify explosive particles in latent fingermarks (321). Fingertips were first 

contaminated with explosives (i.e., TNT, trinitrobenzene, or ammonium nitrate) by 

touching a glass slide on which explosive powder was spread. Excessive powder 

was brushed away, until no more powder was visible by the naked eye, and the 

fingers were pressed on stainless steel (this substrate is transparent in the infrared 

region) as well as on a more realistic substrate (i.e., metal lid from a jar). The 

procedure consisted in locating (explosive) particles (as small as 20 µm) and 

comparing their FTIR spectrum with an infrared spectral library. It should be noted 

that sweat residue could interfere with the resulting FTIR spectrum. Similarly to this 

work, the use of RDX-containing solution were used to contaminate fingertips before 

they were pressed on aluminium coated slides, and further analyzed using ordinary 

FTIR (322). 

The use of a reflective ultraviolet imaging system (RUVIS) is reported to locate 

fingermarks on post-blast material (324). The author stated that people generally look 

for fingermarks on tape and batteries (used to set up the explosive device), but not 

necessarily on the device itself, assuming that the heat of explosion had a strong 

detrimental effect on fingermarks. In this experiment, explosive devices were 

designed using C4, nitromethane and gasoline, a bag and plastic bottles. Following 

the blast in open air, pieces of the explosive device were collected and observed 

using a RUVIS. A single fingermark – further identified to the designer of the 

explosive devices - was found on a non-porous surface (a remaining piece of a 

plastic bottle). The advantage of RUVIS is that the resulting contrast is not dependent 

on the colour of the substrate. Moreover, this technique can be used on-site if 

explosive devices have to be destroyed in place, without a subsequent laboratory 

examination capability (e.g., Irak). 

2.2.15 Detection and lifting of fingermarks on skin 

The detection of marks on skin has been investigated through different 

environmental parameters (325) and by comparing several fingermark 

detection techniques (237, 325). Swedish Black, Magnetic Jet Black 

powder, or RTX spraying are recommended for use in this context (237). 

The lifting of unprocessed marks from skin should be done by using 

Transparent Instant lifter (237), whereas powdered marks should be lifted 

by using silicone casting material (325) or white gelatin lifter (326). 

Four fingerprint powders (i.e., Magnetic Jet Black, Magnetic Silver, Silver Special, 

and Swedish Black) and two chemical methods (i.e., cyanoacrylate fuming and 

ruthenium tetroxyde - RTX) were compared to assess their ability to detect 

fingermarks on skin (237). The marks were either processed directly on skin, or lifted 

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prior to being processed. It should also be noted that very fresh marks were 

considered (i.e., 45 min on living subjects and up to 180 min on dead bodies). Best 

results were obtained with Swedish Black and Magnetic Jet Black powders, as well 

as with RTX spraying. The authors failed to develop quality marks with 

cyanoacrylate, certainly due to the condensation that was formed on the dead 

bodies. 

Four fingerprint powders (i.e., granular black, Lightning grey, white powder, and 

Magnetic Jet Black) were tested to detect fresh and sebum-enriched fingermarks 

deposited on pig skin (325). The Magnetic Jet Black powder was determined as 

being the most suitable powder to use on fresh marks. The author also studied the 

effect of time and skin temperature on the rate of success of fingermark detection. 

However, given that only one mark per temperature value was considered and that 

the pig skin began to putrefy after 43 hours, serious discrepancies were observed 

between results. Consequently, the conclusions of this study (i.e., 43h and 48°C as 

maximal values beyond which no identifiable fingermarks were recovered) should be 

considered with caution. 

Four different lifters (Transparent Instant Lifter, Mipofilm for microtrace, Fuji glossy 

paper and silver plate.) were used for the transfer of untreated fingermarks from the 

skin before being processed (237). Best results were obtained using the Transparent 

Instant lifter. 

Five different lifting methods (i.e., white instant lifter, white gelatin, black gelatin, 

silicone casting material, and transparent adhesive tape) were compared to 

determine which one was the best to lift powdered fingermarks on skin of living 

bodies, using Swedish Black powder (326). Silicone casting material and white 

gelatin have given the best results, on the contrary to white instant lifter and 

transparent adhesive tape which gave the poorest results. 

Baran compared three different lifting materials (i.e., Mastercraft Silicone, AccuTrans, 

and Zhermack High Definition) to collect powdered fingermarks (using Magnetic Jet 

Black) on pig skin (325). The clear Mastercraft silicone casting material constituted 

an efficient solution in that context. 

2.2.16 Miscellaneous detection techniques 

This section reports all the fingermark detection techniques that were 

proposed in the literature during these last three years and which do not fit 

in the previous sections. Some of those deal with emerging reagents or 

procedures, pending further research before considering a casework 

application. Briefly, we can cite: use of a powder-based fire extinguisher to 

detect fingermarks on non-porous surfaces (327, 328), application of heat 

on paper leading to naturally luminescent fingermarks (303, 329, 330), 

polymerization of volatile sulphur nitride on various substrates (331, 332), 

application of a hydrophobic and luminescent gel on non-porous items 

leading to luminescent fingermarks (333), tests of new luminescent 

chemical reagents (334, 335), determination of the best procedures to 

detect oil-contaminated fingermarks or fingermarks left on oily substrates 

(336), successful detection of fingermarks on snakes to fight against 

275

animal smuggling (337), determination of the best procedures to detect 

marks on untreated plywood surfaces (338) or on a petroleum jelly jar 

(339). 

Dean reported the use of dry powder coming from a fire extinguisher to detect latent 

fingermarks on metal, plastic, glass and melamine substrates (327). Such powder is 

composed of ammonium sulphate, monoammonium phosphate, attapulgite clay, 

potassium aluminium silicate, silicon dioxide, methyl hydrogen polysiloxane, and 

colour pigment. The procedure consisted in spraying for a few seconds the 

substrates using a fire extinguisher with a rubber hose extension, then puffing away 

the excess of powders using an air stream and eventually photographing the visible 

fingermarks. The obtained marks are extremely labile and the use of any means to 

enhance the result (such as a brush) would degrade the marks. Lifting of the marks 

also seems to be a difficult task: tape lift or gel lifter should be applied directly on the 

mark and lifted straight up (no attempt should be made to remove air bubbles). 

Apparently, fresh marks from six donors were used, but no comparisons with 

classical detection techniques were illustrated to assess the actual quality of drypowdered 

fingermarks (obtained using a fire extinguisher). Nevertheless, the author 

indicates that his method could be used in the case of a complicated crime scene for 

which traditional methods would be too expensive or time-consuming (328). 

The thermal development (i.e., detection of latent fingermarks on untreated 

substrates, using heat only) has been known since 1940s but has never been really 

applied for casework. It has recently been revisited by some authors (303, 329, 330). 

Brown et al. observed that a short heating (i.e., 10 seconds) of paper substrates at 

300 °C led to UV-fluorescent (but invisible) fingermarks, which become visible (but no 

longer luminescent) after longer exposition (i.e., 20 seconds) (329). The marks were 

characterized by dark brown ridges on light brown background. Excessive heat led to 

a loss of the ridge details and contrast as the papers turned black and combusted. 

For this experiment, sebum- and eccrine-rich fingermarks from ten donors were left 

on various paper substrates, and aged for 48 hours before being processed. Optimal 

observation conditions were determined as being 505 nm for the excitation source, 

and 549 nm for the observation filter. The heat source also seemed to play a major 

role as the heat should be precisely monitored. Under these conditions, the use of an 

oven or a furnace set at 300 °C is recommended, rather that a hot air gun which 

gives uneven heat distribution and can only process a small area at a time. Direct 

contact heating (e.g., by using a heat press or a domestic metal iron) are not 

recommended since such apparatus failed in detecting fingermarks due to a quick 

background colouration. Nevertheless, in a further study, these authors reinvestigated 

the direct contact heating and refined it by using a coated, non-metallic 

or ceramic surface (i.e., a commercial hair straightener) set at ca. 230°C (330). By 

following this procedure, they produced superior results compared that what was 

obtained using hot air, since they succeeded in obtaining both visible and 

luminescent marks (which was impossible to obtain using hot air). The effect of heat 

on the chemical techniques commonly used to detect fingermarks on paper was also 

investigated. Using depletion series (of assumedly fresh fingermarks), thermal 

development was compared with ninhydrin and both techniques were shown to be 

equally sensitive. In an attempt to place thermal development in a sequence, it was 

determined that it should be placed before DFO and ninhydrin, and strictly up to the 

fluorescent stage only (short heating time). Further work is still required to study the 

276

effect of heat on DNA. During their work on evidence recovered from arson scenes, 

Dominick et al. investigated the effect of heat on the natural fluorescence of 

fingermarks on paper (303). They observed that eccrine sweat was most subjected to 

fluorescence between 130 to 180 °C, with a maximum intensity obtained for the 160- 

180 °C range. Optimal observation conditions were determined as being 352-509 nm 

for the excitation source, and 510 nm for the observation filter. It should be noted that 

Brown et al. (329) concluded that heating at lower temperature (i.e., below 200 °C) 

for longer durations did not yield successful fingermark detection, which seems to be 

in contradiction with Dominick's observations. It certainly emphasizes the fact that 

thermal development is still not entirely understood and that further researches are 

required. For example, according to Brown et al., the luminescence arises from the 

degradation of the substrate, while Dominick et al. speculated that it is the result of 

the chemical degradation of the secretion residue due to heat. 

Volatile sulphur nitride (S2N2) is reported to form a dark blue polymer on fingermarks, 

allowing their visualization after two or three hours of exposure (331, 332). Once 

processed, the fingermarks remain visible for several days before the polymer 

hydrolyzes (if exposed to air). The method is said to be successful on a great variety 

of substrates: porous (e.g., paper), non-porous (e.g., glass, plastic wrap, metal), 

cotton, fired cartridges, wetted substrates, washed with organic solvents, or heated 

up to 500 °C for several hours. Nevertheless, the production of S2N2 from S4N4 

seems to be a time-consuming operation requiring adapted laboratory equipment. 

Safety considerations have also to be kept in mind, given that those chemicals are 

sensitive to friction. The use of an adapted sealed vessel is also necessary to 

process fingermarks, due to health and safety issues related with the use of sulphur 

nitride. Besides the detection of fingermarks, it should be noted that S2N2 is also able 

to detect traces of inkjet inks (e.g., traces transferred from a printed paper to an 

envelope, by contact). 

Kwak et al. reported the use of a solid, highly fluorescent, and hydrophobic polymer 

gel able to detect marks by direct contact with a non-porous surface bearing latent 

marks (333). The detection process is based on the increase of the polymer 

fluorescence (more than ten times the initial level) when oily components (from the 

latent secretion) diffuse into the gel film, through close contact. Once transferred into 

the film, the mark can be imaged using UV light. The film can be subsequently rinsed 

by using acetone, which has for effect to “erase” the transferred mark and reinitialize 

the film for further detection. 

Plater et al. proposed new fingermark detection reagents. First, N-alkyl- and N-arylsubstituted 

benzimidazole-2-carboxyaldehydes were tested (334). When these 

reagents are solubilised in dichloromethane, fingermarks on a silica plate were 

detected by immersion, followed by a two-minute-development at 60-100 °C. Upon 

excitation at 254 nm, ridge details could be observed. In another study, 3,7dibromophenothiazin-5-ium 

perbromide was tested as an amino-acid reagent leading 

to a red-coloured product (335). When this compound is dissolved in a 2butanol:pyridine 

(100:0.5 v/v) mixture, fingermarks on a silica plate were detected by 

immersion, followed by a five-to-ten-minute-development at 100 °C. Fingermark 

ridges appear pink or pale red, in visible light. 

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A Master Thesis aimed at determining the best ways to detect fingermarks 

contaminated with oil or deposited on greasy/waxy surfaces was published (336). 

Three types of oil were considered: edible soybean oil, engine oil, and diesel fuel. 

RUVIS was determined as being the most suitable optical method for detecting 

marks on glass. After optical detection, the samples were frozen using liquid 

nitrogen, and kept at -20 °C, before being cyanoacrylate-fumed (and observed using 

RUVIS again). The most suitable subsequent treatments (following cyanoacrylate 

fuming) depend on the kinds of substrates and oils: black powder dusting (followed 

by adhesive lifting) for glass, aluminium, and galvanized iron surfaces; Basic Yellow 

40 staining for glass and aluminium surfaces (this can be followed by black powder 

dusting and lifting); magnetic powder followed by Tex-Lift (a special glue used to lift 

powdered fingermarks from rough surfaces - manufacturer: SPEX Forensics) for 

plastic oil container. It should be noted that Sudan Black, which has been 

successfully tested by the HOSDB (Home Office Scientific Development Branch) for 

such specific oil-contaminated fingermarks, has not been considered in this work. 

In the context of illegal animal smuggling, the possibility to detect fingermarks on the 

skin of reptiles has been investigated (337), the aim being to link the animal with a 

smuggler. Using 48 reptiles of different kinds, fresh sebaceous fingermarks were 

deposited then powdered using a black or white commercial powder. On frozen dead 

reptiles, fresh sebum-rich fingermarks were deposited then cyanoacrylate-fumed, 

and subsequently dye-stained. The authors showed that it was possible to detect 

fingermarks on reptile skin, with some limitations (besides casework conditions, 

instead of fresh sebum-rich marks): it can be difficult to detect marks on some 

reptiles with smaller scales or rough-textured skin. The author suggested the 

possibility to swab the reptile skin to collect DNA samples. 

A thorough study has been conducted about the most suitable techniques to detect 

fingermarks on untreated plywood surfaces (338). For this study, three donors (good, 

average, and poor) were asked to deposit fingermarks on an untreated plywood 

surface. The fingermarks were aged (i.e., from one day to 28 days) before being 

processed by 13 commonly used detection techniques (e.g., powders, ninhydrin, 1,2indanedione, 

cyanoacrylate). Ninhydrin and physical developer (PD) succeeded in 

detecting 28-day-old fingermarks. Iodine fuming followed by 7,8-benzoflavone 

succeeded in detecting one- and three-day-old marks. It was however less efficient 

than silver nitrate, able to detect one- to 14-day-old marks if UV exposure 

immediately follows the silver nitrate processing, to minimize unwanted background 

staining. Nevertheless, on a simulated casework example, it appeared that PD 

applied as a single method (i.e., not integrated in a sequence with ninhydrin or silver 

nitrate) gave the best results in terms of mark recovery rates for untreated plywood 

surfaces. If the result of a PD process is too weak, the use of bleach could help 

enhance the results by darkening the existing marks. 

When facing a sexual assault case, it is possible that a petroleum jelly jar has to be 

processed. Given that such non-porous items (side and lid of the jar) can be 

contaminated with semi-solid petroleum jelly medium, a study investigated the 

possibility to optimize a detection sequence, as well as casting methods (339). For 

this study, one donor was asked to rub his fingertips on petroleum jelly before 

depositing fingermarks on jars (side, lid, and label). The fresh marks were processed 

according to eight different sequences, using five common detection techniques (i.e., 

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cyanoacrylate fuming, sudan black, magnetic powder, ninhydrin, and physical 

developer). Different casting materials were also tested, as well as two storage 

conditions (i.e., room temperature and freezer). The best method consisted in 

processing the item at room temperature, according to the following sequence: first 

photographing visible prints using oblique light, then casting with Mikrosil before 

applying sudan black (this method will colour the underlying label, making the 

fingermark appear as white ridges on a darker background). 

2.3 Forensic light sources, photography and digital/chemical imaging 

2.3.1 Alternative light sources (ALS) 

Nd-YAG laser (340) and, for the first time, LED arrays (341) were 

proposed as alternatives to the commonly used alternative light sources 

based on xenon arc or quartz halogen lamps. The use of an ultraviolet 

emitting light source combined with a specific digital recording was also 

described (324, 342). Finally, the forensic application of infrared imaging 

was discussed (343). 

Lin et al. discussed the forensic application of infrared imaging (343). A digital 

camera able to record infrared light combined with a filter blocking all wavelengths 

below 930 nm has been used to capture bloodstains (dilution series), inks on fabric, 

tire marks, gunshot residue, and charred document on dark background. Even if this 

technique has not been applied to detect fingermarks, it could be interesting to note 

that bloodstains were shown to absorb the near-IR light, and could be detected up to 

1/4 dilution when using white light, and up to 1/8 dilution when using an IR light 

source. 

Following their studies on the use of a Nd-YAG laser in the ultraviolet region to detect 

sebum-rich fingermarks (284), Akiba et al. reported that a 280 nm excitation source is 

optimal to visualize fluorescent marks on white paper, with emission peaks at 330 

and 440 nm (340). Under continuous excitation, the fluorescence at 330 nm 

decreases while the one at 440 nm increases. It should be noted that this technique 

seems to be successful on sebum-rich marks, but gives poor results on eccrine-rich 

ones. 

Besides the conventional ALS based on xenon arc lamps or quartz-halogen lamps 

with filters, Takatsu et al. proposed an alternative based on LED arrays (341). LED 

arrays present two major advantages: (1) the lower cost compared to traditional ALS, 

and (2) the uniformity of the emitted light throughout the illuminated area. One of the 

biggest disadvantages is certainly the output power of an individual LED that is 

insufficient to detect weakly luminescent marks. To overcome this problem, arrays of 

several LED units were considered (i.e., 50 x 60 mm arrays composed of 90 LED). 

Nevertheless, the illumination intensities of LED arrays are still weaker by a 

proportion of 1:7 to 1:9 compared to a xenon arc lamp such as the Polilight. The 

authors tested two kinds of LED arrays (i.e., blue and green) on dye-stained 

cyanoacrylate-fumed fingermarks left on plastic, and on metal-complexed ninhydrinprocessed 

fingermarks left on paper. Well-contrasted images were obtained, making 

of LED-based excitation systems an attractive alternative for those for which 

conventional ALS are too expensive. 

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The use of reflected ultraviolet (such as RUVIS) is recommended as an efficient 

optical method allowing the detection of marks before any chemical treatment or in 

specific occasions (e.g., bitemarks or post-blast evidence) (324). However, one of the 

difficulties inherent to this technique is the photographic recording of the detected 

marks since reflected ultraviolet is invisible to the eye. Sanfilippo et al. discussed this 

issue and gave an overview of the advantages related with the use of reflected UV, 

as well as some experimental recommendations for a successful digital recording 

(342). First, they recommended to replace old camera loaded with panchromatic 

films with a recent digital camera able to record in the ultraviolet region (without the 

need of a light intensifier) and equipped with a “live-view” system, such as the Fujifilm 

FinePix S-3 Pro UVIR or IS-Pro. The sensitivity of the current sensors is sometimes 

better in the infrared region than in the near-ultraviolet light. To circumvent this 

problem (that could cause an unwanted loss of contrast), the use of highly efficient 

filters able to block all non-UV radiations (i.e., visible and infrared radiations up to 

1,100 nm) is recommended in replacement of the classical barrier filters (which might 

let pass some near-infrared light). The authors cite, for example, the Baader Venus 

filters. A less efficient alternative could consist in stacking two classical barrier filters, 

as a way to maximize the blocking of all non-UV radiations. Finally, the light source 

used to illuminate the evidence has a major role to play, since a pure near-ultraviolet 

light is preferred to an incandescent light (that emits near-infrared light as well). 

2.3.2 Photography 

When facing multicoloured backgrounds, full benefit should be taken from 

digital imaging functions, such as high dynamic range (HDR) (344) or the 

Black & White adjustment function included in the Adobe Photoshop 

software (345). When dealing with fingermarks left on rear-view mirrors, it 

could be necessary to remove the back reflective layer before attempting 

to capture the marks (346). Finally, a discussion about the image quality is 

proposed (347). 

The issues related to the digital recording of cyanoacrylate-processed fingermarks 

found on the interior of rear-view mirrors (i.e., inside vehicles) has been addressed 

(346). Indeed, due to the double reflection effect caused by such mirrors, ridge 

patterns may be split into two overlapping ones, creating fuzzy areas reducing the 

overall quality of the mark (even using coaxial lighting). To overcome this problem, 

the author proposed to remove the reflective layer at the back of the mirror by using a 

paint remover and cleaning abrasive metal oxide. 

Day proposed a tutorial explaining the advantages of using high dynamic range 

(HDR) images to extend the luminance of some fingermark images (344). An HDR 

image is built through the fusion of different photographs of the same scene but 

taken with different exposure times. Such operations are now easily achieved in 

some commercially available imaging software, such as Adobe Photoshop. This 

technique could be of interest in the case of scenes with strong differences between 

dark and bright regions, or presenting difficult contrast conditions (e.g., powdered 

mark on a soda can or ninhydrin-processed fingermark on a pink paper). HDR 

processing allows producing ridge detail of greater quality and contrast compared to 

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normally exposed images subsequently processed with standard digital 

modifications. 

When speaking about fingermarks (or about reference fingerprints), the image quality 

is of a first importance if it is defined as the apparent visibility of the existing ridges 

and ridge details. This subject is discussed by Smith in an article whose aim is to 

encourage the readers to think about such issues (347). 

A tutorial was dedicated to the use of a function named “Black & White adjustment” 

(B&W), included in the Adobe Photoshop CS3 software (and later), to enhance 

images of fingermarks left on multicoloured substrates (345). The B&W option 

provides the ability to work on a monochromatic representation of an image while still 

having the full access to the individual colour channels. Compared to the “channel 

mixer” function, which works on three colour channels (i.e., red, green, blue), the 

B&W option allows to work on six colour channels (i.e., red, green, blue, cyan, 

magenta, yellow), providing fine image tuning capabilities such as multicoloured 

background removal. 

2.3.3 Chemical imaging using Fourier transform infrared (FTIR) and Raman 

FTIR 

Vibrational spectroscopic techniques, such as Fourier transform infrared 

(FTIR) or Raman, represent a non-invasive way to investigate the 

chemical composition of latent fingermarks left on non-porous (348, 349) 

or on difficult substrates (350, 351), as well as to monitor the presence of 

exogenous substances, such as cosmetics (352), drugs (352-356) or 

explosives (320-322, 353, 357). But they could also be used to image 

superimposed fingermarks (357). Nevertheless, the sensitivity of these 

spectroscopy-based methods (to image fingermarks or to identify 

contaminants) strongly depends on the differentiation between the 

spectrum of the material from the spectral response of the substrate and 

of the secretion. Moreover, the issues related to the size of the imaged 

area, the difficulty to localize a mark on an entire item prior to any imaging 

attempt, and to the scanning time still constitute limiting aspects that are 

not always sufficiently emphasized in the contributions. 

The conjunction of attenuated total reflection microscopy with Fourier transform 

infrared spectroscopy (ATR-FTIR) provided the possibility to image gelatine-lifted 

(untreated) fingermarks to study the chemical composition of various components 

contained in adult fingermarks (348, 349). The advantage of ATR-FTIR, compared to 

conventional FTIR, mainly lies in the possibility to reduce the interference of the 

substrate (the gel lifter in this case) that could cover and hinder the spectral features 

of the latent secretions. In a first study, sebum-rich fingermarks were left on various 

substrates (e.g., door handle, mug handle, curved glass surface, computer screen) 

before being lifted using two commercial gel lifters (i.e., BVDA Gelatine Lifter and 

Dycem Gel Print Lifter) and analysed by ATR-FTIR. Imaging of sebaceous-rich 

regions of the lifted-fingermarks showed that the gels could spectrally dominate and 

hinder the latent secretion spectra, making difficult the investigation of the chemical 

composition of the fingermarks. By changing the angle of incidence, the interference 

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from the gel could be decreased and ridges could be visualized. The authors 

recommended the BVDA Gelatine Lifter to be used on this kind of imaging. In a 

second range of experiments, the authors imaged fingermarks left on non-porous 

surfaces and heterogeneously contaminated with cosmetic and drugs, by following 

the same procedure (352). Fingermarks were obtained by either rubbing the donor's 

head which had previously been treated with cosmetics, or by first touching drug 

powder (i.e., gamma-hydroxybutyrate - GHB) before depositing, lifting, and imaging 

the marks. They finally tried to image cosmetic-contaminated marks on fabric (cotton 

and polyester) and imaging them directly (without lifting them). Even though it was 

not possible to reconstruct the pattern of the fingermarks, the cosmetic products left 

on the clothes were successfully identified. The equipment allowed imaging an area 

of a size of 2.5 x 3.6 mm² (348) or 4.3 × 5.9 mm² (352), with a total scanning time of 

ca. 100 seconds per image. 

Other studies investigated the use of FTIR to image latent fingermarks contaminated 

with explosives (321, 322), as already described in the “CBRNE-related evidence” 

section. 

Ordinary reflectance FTIR was chosen to image processed fingermarks left on 

“difficult” surfaces, that is, containing illustrations or text that render the visualization 

difficult (e.g., polymer banknotes, papers, masking tapes, or aluminium drink cans) 

(350). An image area up to 4.48 x 4.48 cm² could be scanned through mosaics, in a 

few hours. One of the key-points of the study consisted in a systematic optimization 

of the settings (e.g., spectral resolution or number of scans) so that the collection 

time and the file-size were reduced without compromising the spatial resolution and 

the quality of the final fingermark images. For this study, sebum-rich fingermarks 

were left on various substrates before being processed by cyanoacrylate fumes or 

amino-acid reagents (i.e., ninhydrin, DFO, or 1,2-indanedione), then imaged by FTIR. 

Cyanoacrylate-fumed fingermarks on non-porous substrates yielded good results, 

especially when imaging the frequency slice within the 1750–1800 cm -1 range. On 

the contrary, the attempts to image ninhydrin-processed fingermarks on paper failed 

in giving positive results due to the swamping effect of the cellulose constituents of 

the paper. Moreover, the amount of product formed in fingermarks developed by 

ninhydrin or DFO must be less than or equal to the original number of amino acid 

molecules, whereas cyanoacrylate development of fingermarks has the advantage of 

amplification (which is of a first interest during the imaging step). The authors did not 

use ATR-FTIR, which may have enhanced the results, because “ATR imaging of an 

area as large as a fingermark was not technically feasible at the time of writing” 

(citing the authors). 

FTIR was used to locate and identify exogenous substances contained in latent 

fingermarks (e.g., caffeine, cocaine, paracetamol) through comparison with pure 

product libraries (353). For this study, sebum-rich fingertips were put in contact with 

the powder of interest (the excess of powder being brushed away from the finger), 

before being pressed on either a metal oxide-coated glass slide (Kevley 

Technologies) for reflection experiments, or on a silicon window for transmission 

experiments. Full fingermark images were obtained by scanning an overall area of 

1.12 × 1.12 cm, which took ca. 1h20. Different library searching algorithms were 

tested and compared, when trying to identify the exogenous materials. 

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The use of FTIR has also been proposed to detect superimposed or contaminated 

marks on reflective substrates (e.g., door knob, knife blade, or handle) (357). For this 

study, aluminium-coated glass substrates were used to image all the samples, and 

three scenarios were considered: (1) latent marks left by unwashed hands, (2) 

superimposition of two marks left by two differently-washed hands, and (3) latent 

marks left by hands having touched RDX powder. The authors observed that sebumrich 

features formed continuous patterns along the friction ridges, while protein-rich 

material was deposited as flakes along the ridges, producing a speckled pattern. The 

second scenario illustrated that morphological information obtained through imaging 

was similar to what could be obtained by chemical development followed by optical 

imaging, but that additional information could also be linked with the intrinsic 

chemical composition of the secretions. This allowed distinguishing two different 

marks that were optically similar in appearance. The third scenario consisted in the 

spatial localization of material without the need to sweep the entire area (to be further 

analyzed). No precise information was given on the time required to image a sample, 

or on the scanning size (even if sequential moving of the sample allows extending 

the scanned area to the required size). 

Maynard et al. conducted a thorough study about the chemical imaging of latent 

fingermarks in the near-infrared region (NIR) (351). For this study, sebum-rich latent 

fingermarks were left on several substrates (eight porous: cardboard, cheques, 

newsprint, and five coloured office papers – four semi-porous: white and coloured 

glossy inkjet paper, soft drink labels, and Australian banknotes – three non-porous: 

clear polyethylene bags, orange recycled polyethylene garbage bags, and glass 

microscope slides). The marks were allowed to age for one day, one week, two 

weeks, and one month, before being processed by conventional detection 

techniques. The absorption and luminescence properties of the treated marks were 

then examined over the spectral range 650–1100 nm, using a Condor Macroscopic 

Chemical Imaging System. Under these wavelengths, some fingermark detection 

reagents were characterized by NIR absorption (e.g., ninhydrin, iodine/benzoflavone, 

physical developer, powdering) or emission (e.g., 1,8-diazafluoren-9-one, ninhydrin 

with/without metal salt post-treatment, 1,-2 indanedione, genipin, cyanoacrylate 

stained with NIR dyes, dye-coated metal nanoparticles), resulting in a good contrast 

between the marks and some difficult substrates (e.g., highly luminescent). It should 

be noted that most of the good results were obtained on the fresh mark sets, 

especially the NIR absorption imaging. 

Raman 

West & Went applied Raman spectroscopy to the detection and identification of 

exogenous materials present in fingermarks after those were powdered and lifted 

(354, 355). This study is thus not directly related with imaging of the ridge patterns, 

but more on the identification of exogenous materials in fingermarks. In a first study, 

the authors used commercially available analgesic powders, as well as GHB, to 

contaminate fingertips by contact, before leaving fingermarks on glass slides (total of 

100 marks from one donor) (354). In another study, fingertips were contaminated 

with seized drugs (i.e., ecstasy, cocaine, ketamine, and amphetamine) before being 

powdered, lifted, and imaged (total of 70 marks from one donor) (355). Those marks 

were further processed with aluminium or magnetic iron powder, then lifted using 

lifting tape or hinge lifter, and imaged by Raman spectroscopy. If the application of 

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fingermark detection powders prior to the Raman analysis did not interfere with the 

detection and identification of the contaminants, it however increased the time taken 

to locate the exogenous material due to the physical presence of more material 

within the fingermark (four to five fold increase of time). Special care should be taken 

when choosing the lifting tapes since interfering Raman bands coming from the lift 

are to be avoided. Successful imaging of the sample through transparent evidence 

bags was performed, meaning that the items do not have to be removed from such 

bags to be examined. This consequently reduces contamination risks, and allows 

preserving the chain of evidence. 

Emmons et al. used Raman chemical imaging to detect and identify explosives (as 

trace contaminants) in contaminated fingermarks (320), as described in the “CBRNErelated 

evidence” section. 

Raman spectral mapping was used on tape-lifted fingertips to extract chemical 

information about sebum or contamination drug particles (356). For this study, one 

donor was asked to rub his fingers on his forehead, or to touch drug mixtures 

(containing ibuprofen, arginine and sodium bicarbonate for a first one, and sucrose 

and aspartame for a second one), before a Scotch tape was applied directly on his 

fingertips to lift a fingermark. These tapes were further imaged by Raman. Scanned 

areas were of 200 x 200 µm² for sebum-rich marks, and of 400 x 400 µm² for the 

drug-contaminated samples, which is too small to image a whole ridge pattern but 

sufficient to identify the contaminants. Advanced multivariate data analysis of the 

spectra was however necessary to overcome the overwhelming contribution of the 

tape on the spectra, and to be able to identify the trace components (sebum or drug). 

2.3.4 Miscellaneous imaging techniques 

Several other techniques were developed with a common aim: imaging the 

ridge patterns according to chemical or physico-chemical behaviour. 

However, these techniques are still seldom used compared to FTIR and 

Raman, mainly due to (1) the scanned area being too small to image a 

whole ridge pattern, (2) labor-intensive methods requiring too specific 

equipment. Therefore, most of the contributions constitute only preliminary 

studies that may be further investigated before considering casework 

application. These imaging techniques, briefly described below, are based 

on: scanning electrochemical microscope (358-361), electrochromic 

enhancement (362), tomography (363), surface roughness and refractive 

index (364), mass spectrometry (365), time-resolved luminescence (366), 

MALDI-ToF-MS (367), ToF-SIMS (368), scanning probe microscopy (369), 

MALDI-MS (370), and SALDI-ToF-MS (258-260). 

Scanning Probe Microscopy (SPM) was used to analyse latent fingermarks left on 

glass slides (369). The scanned area was 40 x 40 µm², which is insufficient to image 

a whole fingermark. The authors proposed the use of the “step-and-scan” mode that 

permits to enlarge the scanned area by generating a mosaic composed of 40 images 

of 70 x 70 µm². Nevertheless, this is still insufficient to distinguish ridge patterns. 

Attempts to analyse superimposed fingermarks was also illustrated. 

284

Zhang et al. showed that it was possible to chemically image multimetal deposition 

(MMD)-enhanced fingermarks using a scanning electrochemical microscope (SECM) 

(358-360). This method is based on the measurement of the redox activity on a 

localized area through to the solubilisation of silver. Since silver is preferentially 

reduced on the gold nanoparticles, themselves located on papillary ridges, it was 

possible to visualize the ridge details by scanning fingermarks processed by MMD. 

This method could help visualizing classical MMD results on dark or patterned 

substrates. Alternatively, SECM can also be used on silver-stained or benzoquinonetagged 

fingermarks (361). However, SECM imaging needs to be optimized to enlarge 

the scanning area and to reduce the time required to perform a scan before it could 

be applied in casework. 

Full-field swept-source optical coherence tomography was applied to image latent 

fingermarks left on semi-reflecting glass (363). This technique provides information 

about tomography as well as topography of the marks. According to the authors, the 

detection of very poor quality marks as well as old ones, buried beneath dust layers, 

is possible. Reconstructed ridge details are however not as clear as it should be, and 

the scanned area is smaller than the size of a single fingermark. 

The application of desorption electrospray ionization - mass spectroscopy (DESI-MS) 

was described (365). A solvent was electro-sprayed on the surface bearing the 

sample to be analyzed, generating secondary scattered droplets that are further 

evaporated and analyzed by MS for each point of the surface (spatial resolution: 150 

µm, scanned area: 19.2 x 14.8 mm²). This method gives chemical information, 

associated with the ridge pattern of the fingermark. For this study, fingertips were 

contaminated with drugs and explosives in solution, before being pressed on glass, 

paper and plastic. Good ridge details were obtained, also on tape-lifted marks that 

were analyzed by the same technique. 

Time-resolved luminescence imaging of cyanoacrylate-fumed fingermarks stained 

with an europium-based dye could be used to circumvent the problems caused by 

luminescent substrates (366). This technique took benefit of the long-lived europiumbased 

dye luminescence (micro- to milli-seconds) compared to the short-lived 

background luminescence (nanoseconds). The recording of the image has to take 

place a short time after the excitation source has been shut down. As a result, the 

unwanted background luminescence (or security features) is no more visible, and 

fingermarks can be visualized with a good contrast. Illustrations are given for latent 

fingermarks left on aluminium foil and plastic with security features. 

A diffractive glossmeter sensor is presented as a new imaging technique to visualize 

latent fingermarks left on glossy, curved objects (364). This technique gives 

information about the local gloss of the ridges, depending on the local surface 

roughness and refractive index. By rotating the item (i.e., a ballpoint pen in this case) 

in front of the light source, it was possible to analyze the reflected light coming from 

the scanned area. As a result, latent fingermarks were imaged and coloured 

according to the strength of light reflection. 

Time-of-flight - secondary ion mass spectrometry (ToF-SIMS) was applied to image 

and analyze drug-contaminated fingermarks (368). For this study, washed fingertips 

were contaminated with drug powder (i.e., amphetamine, methamphetamine, and 

285

ecstasy) before being pressed on four different substrates (i.e., steel, aluminium, 

brass and glass). Secondary ions mass spectra were recorded from a 500 x 500 µm² 

area, and further coloured according to the emission intensity of particular ions. This 

window size was insufficient to image a ridge pattern, but sufficient to detect and 

identify the contaminants. 

Matrix assisted laser desorption / ionisation - mass spectrometry (MALDI-MS) was 

used to image the endogenous lipids present in latent fingermarks, as well as their 

degradation products as the marks aged (370). For this study, sebum-rich and 

eccrine-rich fingermarks were left on pre-coated aluminium sheets (thin-layer 

chromatography plates from which the silica coating was scraped off with acetone). 

Ageing of the fingermarks was simulated by placing the samples at 4 °C, 37 °C and 

60 °C for seven days before being processed (i.e., spray-coated with a matrix) then 

imaged. This technique permitted to scan an area of 150 x 150 µm² that can be 

extended to the size of a fingermark (ca. 1.7 x 2.0 cm²) through mosaics, in ca. 44 

minutes. The advantage of chemical imaging being the possibility to visualize specific 

compounds according to their distribution in the sample, many endogenous lipids 

were observed (e.g., cholesterol, palmitoleic acid, stearic acid, or oleic acids). Aged 

fingermarks were imaged by retrieving the oleic acid and its degradation products. 

The authors showed that the matrix used to coat the fingermarks prior to their 

imaging can be eventually removed, while keeping the ability to detect the 

fingermarks using magnetic powder (on aluminium and glass). 

Surface assisted laser desorption / ionisation - time of flight - mass spectrometry 

(SALDI-ToF-MS) was used to image fingermarks from smokers, and more 

particularly by retrieving the peaks related to nicotine and cotinine (258, 259) or 

drugs (260). This study has been described in the “Nanoparticles (nanopowder and 

nanocomposites)” section. 

Beresford & Hillman proposed a method based on the selective deposition of an 

electrochromic polymer (i.e., polyaniline) to image fingermarks left on a metallic 

surface (i.e., stainless steel) (362). The technique is based on the fact that the 

electrochemical deposition process is inhibited by the latent secretion (that masks the 

underlying metallic surface), leading to a negative image of the latent fingermarks. 

Sebaceous and eccrine fingermarks were left on stainless steel, aged from two hours 

to 39 days, before being processed and imaged (extraction of images from a filmed 

electrochemical process). It should be noted that even if this technique led to welldefined 

ridge details, a major drawback is to be reported: if the electrochemical 

deposition is too important, the polymer can overfill the grooves between fingerprint 

ridges, resulting in polymer growth extending across the ridges, modifying thus the 

third level details from the ridge borders. 

An attempt to perform chemical imaging on fingermarks detected using sputtered 

gold and matrix assisted laser desorption / ionization – time of flight - mass 

spectrometry (MALDI-ToF-MS) was presented (367). If sputtered gold hits the 

secretion matrix, it is supposed that it prevents the gold aggregation, favouring the 

formation of gold nanoparticles of smaller sizes and pseudospherical shapes. On 

another hand, if gold hits the substrate (e.g., the grooves between the ridges), gold 

nanoparticles of irregular shape and larger size are created. Both kinds of gold 

nanoparticles exhibit different absorption bands in the visible region, leading to a 

286

visible contrast (this principle is highly similar to vacuum metal deposition). Chemical 

information was obtained through the MALDI-ToF-MS equipment. For this study, 

highly sebum-rich fingermarks from one donor were used by pressing the fingertip for 

ten seconds on a substrate. Contaminated fingermarks were also obtained by putting 

the fingertip in contact with drug powder for 20 seconds before pressing it on a 

substrate. Finally, experiments were conducted on superimposed fingermarks by 

artificially enriching each fingermark with a specific lipid (i.e., stearic acid for the first 

one and oleic acid for the second one). Unfortunately, these protocols are far from 

being realistic and degrade therefore the quality of the observations made. 

3 Miscellaneous 

Survival and transfer of bacteria has been tested on three biometric readers 

(Recognition Systems HandKey III, Crossmatch Verifier 300 LC and TechSphere VP- 

II S). Two strains of bacteria were tested independently of each other: Escherichia 

coli and Staphylococcus aureus. For the survivability test, a known concentration of 

bacteria was applied to the devices (after sterilization) and recovered by a single 

touch with a finger covered with a sterile finger cot after 5, 20, 40 and 60 minutes. 

For the transferability test, the device was touched 50 times with a finger, and 

contamination of the device occurred prior to touch number 11. The survivability 

study shows that at five minutes past contamination, 15% of the Staphilococcus 

aureus remain on the fingerprint reader (and 20% and 40 % for the vein and hand 

geometry readers, respectively); after 20 minutes, the survival rate was almost zero. 

For Escherichia coli, the percent survival after five minutes is 28% for the fingerprint 

reader (it is 40 and 50% for the hand geometry and vein readers, respectively). 

Again, after 20 minutes, the survival rate approaches zero. The number of cells 

transferred decreases over successive touches; most of the bacteria are transferred 

within the first ten touches (371) 

The relationship between oiliness, humidity, elasticity of the skin and image quality 

has been investigated using 190 individuals, 6 impressions per individuals and 9 

fingerprint sensors (4 capacitance, 4 optical and 1 thermal). The skin moisture, 

oiliness and elasticity were measured with a Triplesense device. While differences 

appear between the different sensor technologies, the relationship between skin 

moisture, oiliness and elasticity and image quality is not linear (372). 

The important factors in fingerprint scanners with respect to image quality (quality 

being defined with respect to the accuracy of automated fingerprint recognition) have 

been investigated. The main quality parameters defined by the FBI are used, and 

images are acquired and then degraded with respect to each of these parameters. 

No significant drop in performance (with respect to the ideal case) was observed 

when all parameters were set to the IAFIS IQS minimum requirements (but 

acquisition area is not defined); The PIV IQS requirement for the acquisition area 

allows a drop in performance when the minimal requirement is used. The three most 

critical parameters are acquisition area, output resolution, and geometric accuracy. 

When these three parameters were degraded simultaneously, the drop in 

performance was larger than the sum of the individual performance drops. (373) 

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Saijo et al. propose ultrasound imaging of fingerprints for medical purposes. The 

proposed method allows obtaining clear 3D images of the finger surface, but also of 

the rear surface of the fingerprint (dermal papillae), in vivo (374). 

Causin et al. (375) present the analysis of latex gloves using thermogravimetric 

analysis. This yielded a discriminating power of 99.5%. 

Stature estimation from hands and phalanges is described by Habib and Kamal 

(376); these estimations show a good mean correspondence to stature. Right and left 

hand and foot lengths are investigated for a North Indian population. In particular, the 

goal is to be able to infer the size of a missing member for identifying the deceased; 

indeed, these measures are correlated and regression equations are proposed, 

where, for example, the right foot length is predicted from the right hand length and 

breadth (377). In the same context of identification of human remains, a sex 

difference of the index to ring finger ratio is found (on 300 subjects of south Indian 

origin); this ratio is larger for females (378). 

Packaging of objects for the protection of different kinds of marks, including 

fingermarks is described by Tietze and Heer (379); for porous surfaces, paper bags 

or cardboard boxes should be used, for nonporous surfaces friction should be 

avoided; generally again cardboard is privileged, but plastic containers are also 

mentioned (for example for weapons, that are also the object of special attention in 

this article). 

The organization and fitting out of forensic laboratories has been analysed, and a 

basis for the conception of such laboratories has been presented. This concept is 

based on several modules that can be assembled according to the needs of a given 

laboratory. It emphasizes rationalisation, for example through shortening the 

distances to be covered in particular working sequences (380). 

3.1 Knuckles 

Both the inside and the outside of knuckles has been investigated for their use in 

biometric identification. The inside of knuckles, obtained from images of the whole 

palms, has been used by Nanni and Lumini. An equal error rate of a minimum of 0.3 

is obtained for one knuckle, and when both knuckles from middle and ring fingers, 

the equal error rate obtained in the best scenario is of zero, but obtained on 72 users 

only (and 10 images of the right hand for each user) (381). The outside of knuckles 

has been investigated by Kumar and Ravikanth (382), as well as Zhang et al. (383) ; 

they demonstrate that finger knuckles can be used to distinguish same-source from 

different-source acquisitions in a biometric setting. Whether or not this discrimination 

could be achieved when marks (if such marks did occur) are compared to reference 

material is not the subject of these articles, and therefore remains unknown; however 

these are promising biometrics that could help in source attributions. 

3.2 Barefoot impressions 

An overview of research on barefoot impressions has been proposed (384). Maltais 

and Yamashita (385) present an operational trial where nine marks from bare feet 

288

and ninety inked impressions were sent to 15 specialists. There was one erroneous 

exclusion (but the examiner requested better standards). Also, current Canadian 

criteria do not allow positive identification of barefoot impressions ; therefore, several 

inked prints could be included for a given mark. Several examiners therefore included 

inked impressions other than that of the foot that had left the mark. 

A study on an Indian subpopulation who customarily walks barefoot reports different 

measures, such as the frequency of the relative lengths of toes, the presence and 

number of humps in the ball line, creases, and the flatfoot condition (386). 

Stature and weight estimations based on footprint measurements have been 

reported in different studies. Krishan (387) estimates stature from various measures 

carried out on 2080 footprints from a Gujjar population (an ethnic group from North 

India). Stature is also estimated from a different Indian subpopulation, and it is 

highlighted that the relationship between foot length / breath and stature diverges 

between populations (388). Finally, Kanchan et al. also estimated stature from foot 

measurements, based on a sample of 200 subjects belonging to a North Indian 

population (389), a follow-up from previous work (390). 

The relationship between footprints and weight has also been studied. Here, 50 

volunteers (again from the Gujjar population) left footprints normally, and carrying 

5kg and 20kg additional weight (391). This kind of estimation is less precise than the 

estimation of stature from footprints. 

Gait dynamics are the subject of (392). Measures of footprints left both on a flat 

surface and on sand are analysed, as well as the pressure of different foot regions, 

impulses, velocity and 3D kinematics. Finally, the footprint depth in different foot 

regions is correlated with body mass, functional leg length, foot type, velocity, 

pressure and impulse. Interestingly, body mass is not significantly correlated with 

footprint depth, in any of the considered regions. When stepwise regression is used 

by foot region, pressure, leg length, velocity and impulses are the only variables that 

appear (392). 

3.3 Ears and earmarks 

The ear is receiving attention from both biometric and forensic perspectives. The 

advances in the use of ear image in biometric systems are covered by Hurley et al. 

(393) as well as in another review (394). Attarchi et al. (395) propose a new 

mechanism for biometric recognition. Some entries of the Encyclopedia of Biometrics 

(396-398) are also dealing with the biometric analysis of ear marks and shapes. 

Ear length and breadth, as well as a classification of the type of ear (long and narrow, 

short and broad, long and broad), position of the ear with respect to the head, 

whether the ear is vertical or oblique, and the ear lobe types (free, attached, absent) 

have been investigated on a sample of Kanyakuja Brahmin male subjects (399). 

While this data, which is collected from direct measurements and from photographs 

of the ears, cannot be directly applied to casework, variation is shown in these 

different measures. However, the frequencies of the different classes are reported 

separately. 

289

The efficiency of 20 observers to associate ear photographs has been shown by 

Asirdizer et al. on a database of 120 ear images. The error rate reported varies 

between right and left ear and lies between 12% and 15% (400). 

An investigation of the analysis of DNA obtained from earmarks shows that DNA can 

be obtained from these marks, but also that a large number of non-donor alleles are 

obtained. Full donor profiles were obtained from 10 swabs of ears. However, out of 

these 10 analyses, 6 were contaminated with non-donor alleles, the source of some 

of which is not identifiable (401). 

Automated ear identification is proposed and has been tested on two databases; one 

containing four images per person of 77 persons and one containing six images for 

each of 17 persons. On these databases, recognition rates of 90.50 and 95.05% are 

obtained, using 2 and 3 images for training, respectively; in the second database, 

when 4 images were used for training, the recognition rate is of 97.05% (402). 

Using pairwise comparison of 700 ears by measuring 12 distances between 

landmarks defined on the ear and computing the Euclidean distances between these 

measures, it is demonstrated that there is variation between different ears. The 

measures were taken directly on the ear, not on impressions, and 3 of the 

measurements are depths. The repeatability of measurements was tested on 20 ears 

(from 20 volunteers) by repeating the measures 10 times. An intra-pair distance of 

0.251 was therefore defined. All distances for comparisons between different ears 

from males (350 ears) were above this distance, while for female subjects, out of 

61075 paired combinations, 21 pairs of right ears and 26 pairs of left ears yielded 

distances that were inferior to this limit. Comparisons between the right and the left 

ear for all volunteers were also carried out and only few of these comparisons yielded 

distances smaller than the within-variability threshold (1 male or 0.3% and 7 females 

or 2%). For the pairs of ears with distances below the threshold, images were 

superimposed; the pairs could be distinguished on this basis (403). 

Measurements carried out on ears (left and right, 414 individuals) showed symmetry 

between left and right ear for height and width. A difference between males and 

females was observed for the mean axis of the pinna, and a decline in anti-helical 

take-off angle was observed with increasing age for male and female Caucasians 

(but not for the Afro-Caribbean or Indian subcontinent ethnic groups). Length and 

width of ear increased for all ethnic groups with age. Also, volunteers from India had 

the longest ears, followed by Caucasians and Afro-Caribbeans ; the relationship is 

only significant for males (404). Differences between males and females as well as 

between age groups (linear distances, areas and symmetry increase with age, while 

sagittal angles of the auricle and width to length ratios decrease with age) was also 

found in a study on 843 volunteers (405). 

3.4 Lip marks 

A historical review of lip print studies, a description of the anatomical aspects of lips, 

the different classifications of lip prints, methods of recording lip prints (including 

latent marks), and problems with the comparison of lip prints are presented by 

Caldas et al. (406). The problems in the comparison are essentially linked to the 

intravariability of such impressions, due to the mobility of the lips, and various 

290

pathological conditions that render impossible this comparison. In the same article, 

the study of palatal rugae is also reviewed in the same way. A study on the patterns 

of lip-prints of 966 Saudi subjects, including 13 identical twins, has been carried out. 

They were then classified according to the Renaud classification system, which was 

slightly modified. No two patterns were found to be identical, including in the identical 

twins (although twins showed some similarity in the groove types). Detailed 

frequencies of the different classes of lip patterns are reported, by sex and for each 

of six regions of the lips (407). Direct measurement of the lips of 532 male and 386 

female subjects has been used to investigate the variability between the dimensions 

of the mouth and lips of male and female subjects, and the age-related differences. A 

relatively large number of measures were carried out in the course of this study; it 

was shown that all volumes and the total lip area, as well as mouth and philtrum 

widths and total lip heights were larger for men than for women. The vermillion height 

to mouth width ratio is larger for girls than for boys and for women than for men, but 

larger for adolescent boys than for girls. All measurements significantly changed with 

age, and there were significant interactions between age and sex (408). 

The development of lipmarks left with lipstick on human skin has been investigated. 

A single type of protective lipstick has been used in this study, applied to a mould 

which was used to leave marks on the skin of 40 deceased individuals, on the right 

side of the neck and the anterior region of the forearm. Observation using a UV light 

source, dusting using REDescent Fluorescent Latent Prints Powder and Nile Red 

and observation under UV were used. Development was carried out 15 minutes after 

having left the lip mark. Observation under UV allowed locating the lip marks. Of the 

two reagents tested, REDescent Fluorescent Latent Prints Powder developed all 

marks, but not Nile Red (409). 

Barbaro et al. investigated DNA typing from lipstick prints on skin. The sample 

consisted of four women, who kissed the face of one man, once with light protective 

and once with long-lasting lipstick. Overall eight swabs were collected. Analyses 

were carried out both using Identifiler and Minifiler kits. Better recovery was obtained 

from light protective lipstick than from long-lasting; however generally a mixture 

between the female and male components was obtained, but both cases where only 

the male component was present and cases where only the female component was 

present were observed. In particular, the profile of one of the women never appeared 

(410). 

Note also that lips have been suggested as a biometric modality as well (411). 

4 Crime scenes and case studies 

Reported case studies were dealing with: indented fingermarks under 

paint (412), the voluntary deposition of a toemark on falsified checks to 

mislead the investigation (413), skin removal and conservation (414), the 

role played (or not) by the deprivation level on the recovery of fingermarks 

and DNA (415), voluntary incorporation of “fingermarks” on some CD 

brands (416), fingermarks on a window frame that resisted a washing step 

(417), processing of a self-adhesive stamp using heat, Wetwop and 

ninhydrin (418), fingermarks on foldable items (419), fingermarks found on 

291

the floor after a bank robbery (420), and processing of corpses in bad 

state (421-424), crime scene processing (425), loss of fingerprint 

characteristics due to a cancer treatment (426), and the identification of a 

deceased individual through occupational marks (427). 

The burglary of a clubhouse led to the detection of several fingermarks and palm 

impressions from the point of entry and other disturbed items (412). More particularly, 

a left palm and left little finger marks were recovered from an internal kitchen door 

and identified to an individual who claimed he had not been in this clubhouse for at 

least four years. In fact, it appeared that these marks were indented in the former 

paint coat (when it was not dry yet), and despite the fact that this kitchen door had 

been painted white 18 months ago and washed several times since, the marks were 

still visible and could be dusted / lifted due to the indented ridges. 

During an investigation on counterfeit checks, unusually large “fingermarks” were 

found on the front of eight checks (413). As a conclusion, it appeared that these 

marks were actually corresponding to the right toe of the suspect, who left them to 

mislead the investigation. 

The observation of a blood impression (i.e., fingers and palm) on the thigh of a dead 

body has been followed by the removal of the piece of skin. The difficulties related 

with its conservation were discussed (414). 

A study has been performed to determine if there exists a relationship between the 

deprivation level of a neighbourhood in which a crime was committed and the 

recovery of fingermarks and DNA by crime scene examiners (CSEs) (415). Burglary 

and auto crime data for Northamptonshire (U.K.) during a three-year period has been 

used (ca. 25,000 crime activities). No statistically significant relationship was found 

between the deprivation level and the time spent on crime scene or the recovery of 

fingermarks from the crime scene. However, a statistically significant relationship has 

been found for DNA recovery, which is more frequently recovered from less deprived 

neighbourhoods. The authors emitted the hypothesis that in less deprived areas, the 

victims are more forensically aware and are, therefore, more able to articulate to the 

CSEs which items from the scene may be of importance in the subsequent 

investigation. 

Two strictly identical fingermarks were discovered on two different CDs, processed in 

the frame of two different crime cases (416). The explanation was that these marks 

were left by the music production company as part of their brand, and are thus 

digitally included on the CDs during the manufacturing process. 

The same fingermark has been detected and lifted within the frame of two different 

burglary cases, which took place at the same apartment - but days after (417). The 

mark was initially lifted from the interior side of a kitchen window in an apartment. 

The damaged window frame has then been superficially cleaned by a repairman 

(apparently, it was sufficient to make the mark becoming latent again, but not to 

erase it), and the same fingermark has thus been dusted and lifted as another 

burglary took place at the same apartment, days after. 

292

The use of heat (using a hair drier and tweezers) to remove self-adhesive stamps 

from an envelope was used to further process the sticky side of the stamp but also 

the paper underneath the stamp (418). The use of Wetwop on the stamp gave no 

results, but the processing of the envelope with ninhydrin led to the detection of a 

mark lying initially beneath the stamp. It then appeared that the mark was transferred 

from the stamp (after it had touched the palm of the suspect) to the paper, giving a 

mirror image of the print. 

Three cases related to the detection of fingermarks on foldable or torn substrates 

have been reported (419). In one case, folding the ninhydrin-processed substrate 

allowed to join two fingermark halves, that was subsequently inserted as a whole in 

an AFIS search. In a second case, a torn paper sheet was reassembled to infer the 

cutting dynamics performed by the suspect. The last case was about the transfer of 

secretion residue that occurred between two paper substrates put in contact, which 

led to the detection of a pair of mirrored fingermarks. 

The report of a bank robbery for which the suspect entered the bank from the roof, 

through the attic, down to a back room was made (420). When dusting the floor, 

using black powder to detect footmarks, the author found a very well-defined 

handmark (palm and fingers) left on the floor when the suspect supposedly crawled 

around the room. 

Four case reports were related with corpses' identification. First, latex has been used 

to obtain a negative copy of damaged fingertips belonging to corpses in a bad state 

of preservation (e.g., mummy and carbonization) (421). Fingertips were first brushed 

with ether to remove the grease from the skin, before a thin layer (ca. 0.5 mm) of 

liquid latex was spread on the finger using a wooden rounded rod. Once dried (ca. 

after 10 minutes), the latex film was removed and placed over a finger-shaped 

support previously soaked in latex, to be finally used in a fingerprinting identification 

procedure (e.g., inked or photographed). It should be recalled that the record 

constitutes a negative picture of the original print, and will have to be reversed in 

colour and shape before using it for identification. The only restriction imposed by the 

latex technique is that the mummified or charred fingertip must not be previously 

treated with sodium hydroxide, because it prevents the rubber from vulcanizing. A 

second case reported the use of black powder on the skin (i.e., fingertips and palms) 

detached from a cadaver in bad state of conservation (422). The use of black powder 

had for effect to darken the ridges and to create a visual contrast enabling the 

photography of the skin, and its use as reference material. This solution has been 

chosen because an inking process would have caused detrimental effects on the 

fragile skin. Finally, the identification of 60-year-old mummified remains (i.e., forearm 

and hand) using fingerprints and DNA was reported (423). Fingertips were 

rejuvenated by rehydration, before being moulded with silicone rubber, then casted to 

create positive impressions of each fingertip. The identification of a badly 

decomposed body is the subject of (424). A possible name of the deceased was 

available, and ten-print records of this person were on file. One of these ten-print 

records showed clear pore details in the palmprints. Only three level II characteristics 

were found to be in agreement, however, a lot of pore detail was visible on both 

impressions and allowed the identification. In a further approach, the skin was 

cleaned, and three more minutiae could be matched to characteristics present on the 

exemplar. 

293

How to proceed on crime scenes for the preservation and further analysis of 

fingerprints and DNA has been described by Lauer (425) 

The loss of fingerprint characteristics due to the use of Capecitabine (a treatment for 

breast cancer) is reported by Garcia-Saenz and co-workers (426). 

A case where occupational marks were used for individualisation has been 

presented. In this case, unusual symmetrical skin lesions were present on the hands 

and the ankle joints of an unknown deceased individual. Based on these lesions, it 

was concluded that the deceased was a coconut tree or coconut palm tree climber; 

this allowed identifying the person (427). 

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(195) J. P. Bono, "Review Of: Challenges to Fingerprints", Journal of Forensic 

Sciences, vol. 55, p. 1137, 2010. 

(196) S. A. Cole, "Out of the Daubert Fire and into the Fryeing Pan?: Self Validation, 

Metaexpertise and the Admissibility of Latent Print Evidence in Frye 

Jurisdictions", Minnesota Journal of Law, Science and Technology, vol. 9, pp. 

453-542, 2008. 

(197) A. Becue, S. Moret, C. Champod, and P. Margot, "Use of Stains to Detect 

Fingermarks", Biotechnic & Histochemistry, DOI: 10.3109/10520290903462838, 


(198) C. Lennard, "Fingerprint Detection: Current Capabilities", Australian Journal of 


(199) R. Jelly, E. L. T. Patton, C. Lennard, S. W. Lewis, and K. F. Lim, "The Detection 

of Latent Fingermarks on Porous Surfaces Using Amino Acid Sensitive 

Reagents: A Review", Analytica Chimica Acta, vol. 652, pp. 128-142, 2009. 

(200) S. Bell, "Forensic Chemistry", Annual Review of Analytical Chemistry, vol. 2, pp. 

297-319, 2009. 

(201) T. Kent, "Standardizing Protocols for Fingerprint Reagent Testing", Journal of 

Forensic Identification, vol. 60, pp. 371-379, 2010. 

(202) R. S. Croxton, M. G. Baron, D. Butler, T. Kent, and V. G. Sears, "Variation in 

Amino Acid and Lipid Composition of Latent Fingerprints", Forensic Science 

International, vol. 199, pp. 93-102, 2010. 

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(203) J. D. Humphreys, G. Porter, and M. Bell, "The Quantification of Fingerprint 

Quality Using a Relative Contrast Index", Forensic Science International, vol. 

178, pp. 46-53, 2008. 

(204) J. Vanderwee, G. Porter, A. Renshaw, and M. Bell, "The Investigation of a 

Relative Contrast Index Model for Fingerprint Quantification", Forensic Science 

International, DOI:10.1016/j.forsciint.2010.05.005, 2010. 

(205) A. Hemmila, J. McGill, and D. Ritter, "Fourier Transform Infrared Reflectance 

Spectra of Latent Fingerprints: A Biometric Gauge for the Age of an Individual", 

Journal of Forensic Sciences, vol. 53, pp. 369-376, 2008. 

(206) K. M. Antoine, S. Mortazavi, A. D. Miller, and L. M. Miller, "Chemical Differences 

Are Observed in Children’s Versus Adults’ Latent Fingerprints as a Function of 

Time", Journal of Forensic Sciences, vol. 55, pp. 513-518, 2010. 

(207) A. Khedr, "The Profile of Free Amino Acids in Latent Fingerprint of Healthy and 

Beta-Thalassemic Volunteers", Journal of Chromatography B, vol. 878, pp. 

1576-1582, 2010. 

(208) V. Kerr, K. Stringer, G. Thompson, N. W. Scott, and M. Fowler, "Testing Latent 

Fingerprint Longevity on Glass after Exposure to Water", Fingerprint Whorld, 

vol. 34, pp. 86-95, 2008. 

(209) I. Soltyszewski, J. Moszczynski, W. Pepinski, S. Jastrzebowska, W. Makulec, R. 

Zbiec, and J. Janica, "Fingerprint Detection and DNA Typing on Objects 

Recovered from Water", Journal of Forensic Identification, vol. 57, pp. 681-687, 


(210) M. A. Wood and I. T. James, "Latent Fingerprint Persistence and Development 

Techniques on Wet Surfaces", Fingerprint Whorld, vol. 35, pp. 90-100, 2009. 

(211) G. De Paoli, S. A. Lewis Sr., E. L. Schuette, L. A. Lewis, R. M. Connatser, and 

T. Farkas, "Photo- and Thermal-Degradation Studies of Select Eccrine 

Fingerprint Constituents", Journal of Forensic Sciences, vol. 55, pp. 962-969, 


(212) R. S. Ramotowski, "Composition of Latent Print Residue", in Advances in 

Fingerprint Technology, 2nd Edition, H. C. Lee and R. E. Gaensslen, Eds. Boca 

Raton, Florida: CRC Press, 2001, pp. 63-104. 

(213) L. Schwarz, "An Amino Acid Model for Latent Fingerprints on Porous Surfaces", 


(214) L. Schwarz and M. Baisel, "Erster Ringversuch Zur Sicherung Latenter 

Daktyloskopischer Spuren Mit Reproduzierbaren Testspurenträgern", 

Kriminalistik, pp. 500-505, 2008. 

(215) X. Spindler, M. Stoilovic, C. Lennard, and A. Lennard, "Spectral Variations for 

Reaction Products Formed between Different Amino Acids and Latent 

Fingermark Detection Reagents on a Range of Cellulose-Based Substrates", 


(216) D. Sapse and N. D. K. Petraco, "A Step on the Path in the Discovery of New 

Latent Fingerprint Development Reagents: Substituted Ruhemann’s Purples 

and Implications for the Law", Journal of Molecular Modeling, vol. 13, pp. 943- 


(217) J. Almog, A. Klein, I. Davidi, Y. Cohen, M. Azoury, and M. Levin-Elad, "Dual 

Fingerprint Reagents with Enhanced Sensitivity: 5-Methoxy- and 5- 

Methylthioninhydrin", Journal of Forensic Sciences, vol. 53, pp. 364-368, 2008. 

(218) J. Almog and H. Glasner, "Ninhydrin Thiohemiketals: Basic Research Towards 

Improved Fingermark Detection Techniques Employing Nano-Technology", 

Journal of Forensic Sciences, vol. 55, pp. 215-220, 2010. 

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(219) O. P. Jasuja, M. A. Toofany, G. D. Singh, and G. S. Sodhi, "Dynamics of Latent 

Fingerprints: The Effect of Physical Factors on Quality of Ninhydrin Developed 

Prints - a Preliminary Study", Science & Justice, vol. 49, pp. 8-11, 2009. 

(220) L. Schwarz and M.-L. Hermanowski, "Luftfeuchtigkeit Als Notwendige 

Bedingung Für Die Daktyloskopische Spurensicherung Mit Ninhydrin: Ein 

Praxisorientierter Und Kostengünstiger Lösungsansatz Mit Kaliumcitrat", Archiv 

fur Kriminologie, vol. 225, pp. 39-45, 2010. 

(221) L. Schwarz, "Daktyloskopische Spurensicherung Auf Saugenden Oberflächen 

Mit DFO: Ein Praxistauglicher Ansatz", Archiv für Kriminologie, vol. 225, pp. 81- 


(222) S. E. Russel, G. L. John, and S. L. Naccarato, "Modifications to the 1,2- 

Indanedione/Zinc Chloride Formula for Latent Print Development", Journal of 


(223) D. E. Bicknell and R. S. Ramotowski, "Use of an Optimized 1,2-Indanedione 

Process for the Development of Latent Prints", Journal of Forensic Sciences, 

vol. 53, pp. 1108-1116, 2008. 

(224) V. Sears, R. Batham, and S. Bleay, "The Effectiveness of 1,2-Indanedione-Zinc 

Formulations and Comparison with HFE-Based 1,8-Diazafluoren-9-One for 

Fingerprint Development", Journal of Forensic Identification, vol. 59, pp. 654- 

678, 2009. 

(225) R. Jelly, S. W. Lewis, C. Lennard, K. F. Lim, and J. Almog, "Lawsone: A Novel 

Reagent for the Detection of Latent Fingermarks on Paper Surfaces", Chemical 

Communications, pp. 3513-3515, 2008. 

(226) J. Chan, R. Shimmon, X. Spindler, P. Maynard, C. Lennard, C. Roux, and B. H. 

Stuart, "An Investigation of Isatin as a Potential Reagent for Latent Fingermark 

Detection on Porous Surfaces", Journal of Forensic Identification, vol. 60, pp. 

320-336, 2010. 

(227) J. L. Lee, S. M. Bleay, V. G. Sears, S. Mehmet, and R. Croxton, "Evaluation of 

the Dimethylaminocinnemaldehyde Contact Transfer Process and Its 

Application to Fingerprint Development on Thermal Papers", Journal of 


(228) M. Stoilovic, C. Lennard, C. Wallace-Kunkel, and C. Roux, "Evaluation of a 1,2- 

Indanedione Formulation Containing Zinc Chloride for Improved Fingermark 

Detection on Paper", Journal of Forensic Identification, vol. 57, pp. 4-18, 2007. 

(229) S. P. Wargacki, L. A. Lewis, and M. D. Dadmun, "Enhancing the Quality of 

Aged Latent Fingerprints Developed by Superglue Fuming: Loss and 

Replenishment of Initiator", Journal of Forensic Sciences, vol. 53, pp. 1138- 

1144, 2008. 

(230) C. McLaren, C. Lennard, and M. Stoilovic, "Methylamine Pretreatment of Dry 

Latent Fingermarks on Polyethylene for Enhanced Detection by Cyanoacrylate 

Fuming", Journal of Forensic Identification, vol. 60, pp. 199-222, 2010. 

(231) S. P. Wargacki, L. A. Lewis, and M. D. Dadmun, "Understanding the Chemistry 

of the Development of Latent Fingerprints by Superglue Fuming", Journal of 


(232) W. R. King, "The Effects of Differential Cyanoacrylate Fuming Times on the 

Development of Fingerprints on Skin", Journal of Forensic Identification, vol. 59, 

pp. 537-544, 2009. 

(233) M. Scott, "Does CA Fuming Interfere with Powder Suspension Processing?", 


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(234) J. Salama, S. Aumeer-Donovan, C. Lennard, and C. Roux, "Evaluation of the 

Fingermark Reagent oil red O as a Possible Replacement for Physical 

Developer", Journal of Forensic Identification, vol. 58, pp. 203-237, 2008. 

(235) M. A. Wood and T. James, "ORO. The Physical Developer Replacement?", 

Science & Justice, vol. 49, pp. 272-276, 2009. 

(236) H. J. Swofford, "The Efficacy of Commercial Vs Noncommercial Physical 

Developer Solutions and the Sequential Enhancement of Friction Ridge Using 

Potassium Iodide", Journal of Forensic Identification, vol. 60, pp. 19-33, 2010. 

(237) M. Trapecar and J. Balazic, "Fingerprint Recovery from Human Skin Surfaces", 

Science & Justice, vol. 47, pp. 136-140, 2007. 

(238) M. Trapecar and M. K. Vinkovic, "Techniques for Fingerprint Recovery on 

Vegetable and Fruit Surfaces Used in Slovenia - a Preliminary Study", Science 

& Justice, vol. 48, pp. 192-195, 2008. 

(239) L. Liu, Z. Zhang, L. Zhang, and Y. Zhai, "The Effectiveness of Strong Afterglow 

Phosphor Powder in the Detection of Fingermarks", Forensic Science 

International, vol. 183, pp. 45-49, 2009. 

(240) S. Mopoung and P. Thongcharoen, "Coloured Intensity Enhancement of Latent 

Fingerprint Powder Obtained from Banana Peel Activated Carbon with 

Methylene Blue", Scientific Research and Essay, vol. 4, pp. 8-12, 2009. 

(241) Q. Chen, W. T. Kerk, A. M. Soutar, and X. T. Zeng, "Application of Dye 

Intercalated Bentonite for Developing Latent Fingerprints", Applied Clay 

Science, vol. 44, pp. 156-160, 2009. 

(242) N. Nic Daéid, S. Carter, and K. Laing, "Comparison of Three Types of White 

Powder Suspensions for the Recovery of Fingerprints on Wetted Nonporous 

Surfaces", Journal of Forensic Identification, vol. 58, pp. 590-599, 2008. 

(243) N. Nic Daéid, S. Carter, and K. Laing, "Powder Suspensions for the Recovery of 

Fingerprints on Wetted Nonporous Surfaces", Fingerprint Whorld, vol. 35, pp. 

122-128, 2009. 

(244) B. J. Jones, A. J. Reynolds, M. Richardson, and V. G. Sears, "Nano-Scale 

Composition of Commercial White Powders for Development of Latent 

Fingerprints on Adhesives ", Science & Justice, 

DOI:10.1016/j.scijus.2009.08.001, 2010. 

(245) A. J. Reynolds, B. J. Jones, V. Sears, and V. Bowman, "Nano-Scale Analysis of 

Titanium Dioxide Fingerprint Development Powders", Journal of Physics: 

Conference Series, vol. 126, art. no. 012069, 2008. 

(246) N. Nic Daéid, S. Carter, and K. Laing, "Comparison of Vacuum Metal Deposition 

and Powder Suspension for Recovery of Fingerprints on Wetted Nonporous 


(247) O. P. Jasuja, G. D. Singh, and G. S. Sodhi, "Small Particle Reagents: 

Development of Fluorescent Variants", Science & Justice, vol. 48, pp. 141-145, 

2008. 

(248) B. J. Jones, R. Downham, and V. G. Sears, "Effect of Substrate Surface 

Topography on Forensic Development of Latent Fingerprints with Iron Oxide 

Powder Suspension", Surface and Interface Analysis, vol. 42, pp. 438-442, 


(249) D. Cohen and E. H. Cohen, "A Significant Improvement to the SPR Process: 

More Latent Prints Were Revealed after Thorough Wiping of Small Particle 

Reagent-Treated Surface", Journal of Forensic Identification, vol. 60, pp. 152- 


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(250) A. A. Cantu, "Nanoparticles in Forensic Science", in Proceeding of SPIE - 

Optics and Photonics for Counterterrorism and Crime Fighting IV. vol. 7119, G. 

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(251) C. Lennard, "Fingerprint Detection: Future Prospects", Australian Journal of 


(252) P. Dürussel, E. Stauffer, A. Becue, C. Champod, and P. Margot, "Single-Metal 

Deposition: Optimization of This Fingermark Enhancement Technique", Journal 


(253) A. Becue, A. Scoundrianos, C. Champod, and P. Margot, "Fingermark 

Detection Based on the in Situ Growth of Luminescent Nanoparticles - Towards 

a New Generation of Multimetal Deposition", Forensic Science International, 

vol. 179, pp. 39-43, 2008. 

(254) D. Gao, F. Li, J. Song, X. Xu, Q. Zhang, and L. Niu, "One Step to Detect the 

Latent Fingermarks with Gold Nanoparticles", Talanta, vol. 80, pp. 479-483, 

2009. 

(255) N. Ul Islam, K. F. Ahmed, A. Sugunan, and J. Dutta, "Forensic Fingerprint 

Enhancement Using Bioadhesive Chitosan and Gold Nanoparticles", in 2 nd 

IEEE International Conference on Nano/Micro Engineered and Molecular 

Systems, Bangkok, Thailand, 2007, pp. 411-415. 

(256) B. J. Theaker, K. E. Hudson, and F. J. Rowell, "Doped Hydrophobic Silica 

Nano- and Micro-Particles as Novel Agents for Developing Latent Fingerprints", 


(257) L. Liu, S. K. Gill, Y. Gao, L. J. Hope-Weeks, and K. H. Cheng, "Exploration of 

the Use of Novel SiO2 Nanocomposites Doped with Fluorescent Eu 3+ /Sensitizer 

Complex for Latent Fingerprint Detection", Forensic Science International, vol. 

176, pp. 163-172, 2008. 

(258) M. Benton, F. Rowell, L. Sundar, and M. Jan, "Direct Detection of Nicotine and 

Cotinine in Dusted Latent Fingermarks of Smokers by Using Hydrophobic Silica 

Particles and Ms", Surface and Interface Analysis, vol. 42, pp. 378-385, 2009. 

(259) M. Benton, M. J. Chua, F. Gu, F. Rowell, and J. Ma, "Environmental Nicotine 

Contamination in Latent Fingermarks from Smoker Contacts and Passive 

Smoking", Forensic Science International, vol. 200, pp. 28-34, 2010. 

(260) F. Rowell, K. Hudson, and J. Seviour, "Detection of Drugs and Their Metabolites 

in Dusted Latent Fingermarks by Mass Spectrometry", Analyst, vol. 134, pp. 

701-707, 2009. 

(261) J. Dilag, H. Kobus, and A. V. Ellis, "Cadmium Sulfide Quantum Dot/Chitosan 

Nanocomposites for Latent Fingermark Detection", Forensic Science 


(262) Y. F. Wang, R. Q. Yang, Y. J. Wang, Z. X. Shi, and J. J. Liu, "Application of 

CdSe Nanoparticle Suspension for Developing Latent Fingermarks on the 

Sticky Side of Adhesives", Forensic Science International, vol. 185, pp. 96-99, 

2009. 

(263) K. H. Cheng, J. Ajimo, and W. Chen, "Exploration of Functionalized CdTe 

Nanoparticles for Latent Fingerprint Detection", Journal of Nanoscience and 

Nanotechnology, vol. 8, pp. 1170-1173, 2008. 

(264) J. J. Liu, Z. X. Shi, Y. Yu, R. Q. Yang, and S. Zuo, "Water-Soluble Multicolored 

Fluorescent CdTe Quantum Dots: Synthesis and Application for Fingerprint 

Developing", Journal of Colloid and Interface Science, vol. 342, pp. 278-282, 


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(265) A. Becue, S. Moret, C. Champod, and P. Margot, "Use of Quantum Dots in 

Aqueous Solution to Detect Blood Fingermarks on Non-Porous Surfaces", 


(266) Y.-J. Jin, Y.-J. Luo, G.-P. Li, J. Li, Y.-F. Wang, R.-Q. Yang, and W.-T. Lu, 

"Application of Photoluminescent CdS/PAMAM Nanocomposites in Fingerprint 

Detection", Forensic Science International, vol. 179, pp. 34-38, 2008. 

(267) Y.-F. Wang, Y.-J. Wang, R.-Q. Yang, and Y.-J. Jin, "Study on Amidation 

Reaction between CdS/PAMAM and Amino Acid and Its Application to Latent 

Fingerprint Development", Spectroscopy and Spectral Analysis, vol. 28, pp. 

2843-2846, 2008. 

(268) G. S. Sodhi and J. Kaur, "A Novel, Nanoparticle Size Fingerprint Detecting 

Composition Based on Fluorescent Lucifer Y Stain", Fingerprint Whorld, vol. 34, 


(269) M. J. Choi, K. E. McBean, P. H. R. Ng, A. M. McDonagh, P. J. Maynard, C. 

Lennard, and C. Roux, "An Evaluation of Nanostructured Zinc Oxide as a 

Fluorescent Powder for Fingerprint Detection", Journal of Materials Science, 

vol. 43, pp. 732-737, 2008. 

(270) K. H. Cheng, J. Aijmo, L. Ma, M. Yao, X. Zhang, J. Como, L. J. Hope-Weeks, J. 

Huang, and W. Chen, "Luminescence Decay Dynamics and Trace Biomaterials 

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Physical Chemistry C, vol. 112, pp. 17931-17939, 2008. 

(271) P. Hazarika, S. M. Jickells, K. Wolff, and D. A. Russel, "Imaging of Latent 

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Chemie, International Edition in English, vol. 47, pp. 10167-10170, 2008. 

(272) P. Hazarika, S. M. Jickells, and D. A. Russell, "Rapid Detection of Drug 

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134, pp. 93-96, 2009. 

(273) M. J. Choi, A. M. McDonagh, P. Maynard, and C. Roux, "Metal-Containing 

Nanoparticles and Nano-Structured Particles in Fingermark Detection", Forensic 

Science International, vol. 179, pp. 87-97, 2008. 

(274) A. Becue, "Les nanoparticules, une nouvelle arme contre le crime?", Actualité 

Chimique, vol. 342-343, pp. 52-58, 2010. 

(275) B. Schnetz and P. Margot, "Technical Note: Latent Fingermarks, Colloidal Gold 

and Multimetal Deposition (MMD) - Optimisation of the Method", Forensic 

Science International, vol. 118, pp. 21-28, 2001. 

(276) V. Drapel, A. Becue, C. Champod, and P. Margot, "Identification of Promising 

Antigenic Components in Latent Fingermark Residues", Forensic Science 


(277) A. D. Reinholz, "Albumin Development Method to Visualize Friction Ridge Detail 

on Porous Surfaces", Journal of Forensic Identification, vol. 58, pp. 524-539, 

2008. 

(278) O. S. Wolfbeis, "Nanoparticle-Enhanced Fluorescence Imaging of Latent 

Fingerprints Reveals Drug Abuse", Angewandte Chemie, International Edition in 

English, vol. 48, pp. 2268-2269, 2009. 

(279) L. Schwarz and I. Klenke, "Improvement in Latent Fingerprint Detection on 

Thermal Paper Using a One-Step Ninhydrin Treatment with 

Polyvinylpyrrolidones (PVP)", Journal of Forensic Sciences, vol. 55, pp. 1076- 


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(280) E. L. T. Patton, D. H. Brown, and S. W. Lewis, "Detection of Latent Fingermarks 

on Thermal Printer Paper by Dry Contact with 1,2-Indanedione", Analytical 

Methods, vol. 2, pp. 631-637, 2010. 

(281) M. Scott, "Improved Results in the Development of Latent Fingerprints on 

Thermal Paper", Journal of Forensic Identification, vol. 58, pp. 424-428, 2008. 

(282) S. G. Varshini Kusenthiran, T. Rogers, and W. Knaap, "Latent Fingerprint 

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Journal of Forensic Identification, vol. 60, pp. 34-44, 2010. 

(283) O. P. Jasuja and G. D. Singh, "Development of Latent Fingermarks on Thermal 

Paper: Preliminary Investigation into Use of Iodine Fuming", Forensic Science 

International, vol. 192, pp. e11-e16, 2009. 

(284) A. Becue, C. Champod, and P. Margot, "Fingermarks, Bitemarks and Other 

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Nic Daéid and M. M. Houck, Eds. Boca Raton, FL: CRC Press LLC, 2010. 

(285) L. Schwarz and I. Klenke, "Enhancement of Ninhydrin- or DFO-Treated Latent 

Fingerprints on Thermal Paper", Journal of Forensic Sciences, vol. 52, pp. 649- 


(286) J. W. Bond, "The Thermodynamics of Latent Fingerprint Corrosion of Metal 

Elements and Alloys", Journal of Forensic Sciences, vol. 53, pp. 1344-1352, 

2008. 

(287) J. W. Bond, "Visualization of Latent Fingerprint Corrosion of Metallic Surfaces", 


(288) J. W. Bond, "On the Electrical Characteristics of Latent Finger Mark Corrosion 

of Brass", Journal of Physics D: Applied Physics, vol. 41, art. no. 125502, 2008. 

(289) J. W. Bond, "Visualization of Latent Fingerprint Corrosion of Brass", Journal of 

Forensic Sciences, vol. 54, pp. 1034-1041, 2009. 

(290) J. W. Bond and C. Heidel, "Visualization of Latent Fingerprint Corrosion on a 

Discharged Brass Shell Casing", Journal of Forensic Sciences, vol. 54, pp. 892- 

894, 2009. 

(291) J. W. Bond, "Determination of the Characteristics of a Schottky Barrier Formed 

by Latent Finger Mark Corrosion of Brass", Journal of Physics D: Applied 

Physics, vol. 42, art. no. 235301, 2009. 

(292) A. J. Goddard, A. R. Hillman, and J. W. Bond, "High Resolution Imaging of 

Latent Fingerprints by Localized Corrosion on Brass Surfaces", Journal of 


(293) E. Paterson, J. W. Bond, and A. R. Hillman, "A Comparison of Cleaning 

Regimes for the Effective Removal of Fingerprint Deposits from Brass", Journal 

of Forensic Sciences, vol. 55, pp. 221-224, 2010. 

(294) S. Johnson, "Development of Latent Prints on Firearms Evidence", Journal of 


(295) K. E. Edmiston and J. Johnson, "Determining an Optimal Sequence for 

Chemical Development of Latent Prints on Cartridge Casings and Shotgun 

Shells", Journal of Forensic Sciences, vol. 54, pp. 1327-1331, 2009. 

(296) D. Molina, "The Use of Un-Du to Separate Adhesive Materials", Journal of 


(297) J. W. Bergeron, "Use of Liquid Nitrogen to Separate Adhesive Tapes", Journal 


(298) D. Cramer and K. Glass, "Developing Latent Fingerprints on the Adhesive Side 

of Tape Using a Freezing Technique", Journal of Forensic Identification, vol. 58, 

pp. 419-423, 2008. 

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(299) G. Bradshaw, S. Bleay, J. Deans, and N. Nic Daéid, "Recovery of Fingerprints 

from Arson Scenes: Part 1 - Latent Fingerprints", Journal of Forensic 

Identification, vol. 58, pp. 54-82, 2008 [also published in: Fingerprint Whorld, 

vol. 35, pp. 12-31, 2009]. 

(300) A. J. Dominick, N. Nic Daéid, S. M. Bleay, and V. Sears, "The Recoverability of 

Fingerprints on Paper Exposed to Elevated Temperatures - Part 1: Comparison 

of Enhancement Techniques", Journal of Forensic Identification, vol. 59, pp. 

325-339, 2009. 

(301) J. Moore, S. Bleay, J. Deans, and N. Nic Daéid, "Recovery of Fingerprints from 

Arson Scenes: Part 2 - Fingerprints in Blood", Journal of Forensic Identification, 

vol. 58, pp. 83-108, 2008 [also published in: Fingerprint Whorld, vol. 35, pp. 64- 

82, 2009]. 

(302) K. L. Tontarski, K. A. Hoskins, T. G. Watkins, L. Brun-Conti, and A. L. Michaud, 

"Chemical Enhancement Techniques of Bloodstain Patterns and DNA Recovery 

after Fire Exposure", Journal of Forensic Sciences, vol. 54, pp. 37-48, 2009. 

(303) A. J. Dominick, N. Nic Daéid, S. M. Bleay, and V. Sears, "The Recoverability of 

Fingerprints on Paper Exposed to Elevated Temperatures - Part 2: Natural 

Fluorescence", Journal of Forensic Identification, vol. 59, pp. 340-355, 2009. 

(304) T. P. B. Larkin, N. P. Marsh, and P. M. Larrigan, "Using Liquid Latex to Remove 

Soot to Facilitate Fingerprint and Bloodstain Examinations: A Case Study", 


(305) S. W. Clutter, R. Bailey, J. C. Everly, and K. Mercer, "The Use of Liquid Latex 

for Soot Removal from Fire Scenes and Attempted Fingerprint Development 

with Ninhydrin", Journal of Forensic Sciences, vol. 54, pp. 1332-1335, 2009. 

(306) J. Moore, S. Bleay, J. Deans, and N. Nic Daéid, "Recovery of Fingerprints from 

Arson Scenes: Part 2 - Fingerprints in Blood", Fingerprint Whorld, vol. 35, pp. 

64-82, 2009. 

(307) G. Langenburg, "Deposition of Bloody Friction Ridge Impressions", Journal of 


(308) J. Zarate, "A Method for Lifting Bloody Impressions Using a Lifting Strip 

Containing Titanium Dioxide", Midwest Forensics Resource Center - Research 

and Development Program Summary, November 2009, pp. 27-29, 2009. 

(309) J. Bergeron, "Development of Bloody Prints on Dark Surfaces with Titanium 

Dioxide and Methanol", Journal of Forensic Identification, vol. 53, pp. 149-161, 

2003. 

(310) J. J. Raymond, C. Roux, E. Du Pasquier, J. Sutton, and C. Lennard, "The Effect 

of Common Fingerprint Detection Techniques on the DNA Typing of 

Fingerprints Deposited on Different Surfaces", Journal of Forensic Identification, 

vol. 54, pp. 22-44, 2004. 

(311) J. Sewell, I. Quinones, C. Ames, B. Multaney, S. Curtis, H. Seeboruth, S. 

Moore, and B. Daniel, "Recovery of DNA and Fingerprints from Touched 

Documents", Forensic Science International: Genetics, vol. 2, pp. 281-285, 

2008. 

(312) J. J. Raymond, "A Criminalistic Approach to Biological Evidence: Trace DNA 

and Volume Crime Offences", in Centre for Forensic Science. Doctorate of 

Philosophy (Science) Sydney (Australia): University of Technology, 2010. 

(313) A. J. Dominick, L. A. Welch, N. Nic Daéid, and S. M. Bleay, "Is There a 

Relationship between Fingerprint Donation and DNA Shedding", Journal of 


312

(314) D. McDonald, H. Pope, and G. M. Miskelly, "The Effect of Chlorine and 

Hydrogen Chloride on Latent Fingermark Evidence", Forensic Science 


(315) R. Hoile, S. J. Walsh, and C. Roux, "Bioterrorism: Processing Contaminated 

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319

  

Biological Evidence 

Review 2007 – 2010 

Reinier J. Verduijn 

University van Amsterdam (UvA) 

  

  

320 

  


1.  ABSTRACT  323  

2.  BIOLOGICAL TRACE EVIDENCE  323  

2.1  Introduction  323  

2.2  Biological traces  323  

2.3  Detection of traces  324  

2.3.1  Fluorescence spectroscopy  324  

2.3.2  Raman spectroscopy  325  

2.3.3  XRF  325  

2.4  Recovery of traces  326  

3.  EVIDENCE IDENTIFICATION  326  


3.2  Blood  327  

3.2.1  Current confirmatory techniques  327  

3.2.2  Emerging and future techniques  328  

3.3  Semen  329  

3.3.1  Current confirmatory techniques  329  

3.3.2  Emerging and future techniques  329  

3.4  Other body fluids  330  

3.4.1  Saliva  330  

3.4.2  Vaginal fluid  331  

3.4.3  Urine  331  

3.4.4  Sweat  331  

3.5  RNA as a tool for evidence identification  331  

3.5.1  mRNA  331  

3.5.2  MicroRNA  332  

4.  EVIDENCE INDIVIDUALIZATION  332  


4.2  DNA typing  332  

4.3  Epigenetics  333  

4.4  Short Tandem Repeats  333  

4.5  Single Nucleotide Polymorphisms  334  

4.6  Polymerase Chain Reaction  334  

  

  

321 

5.  PRESENT CHALLENGES  336  

5.1  Quality Control  336  

5.2  Low Copy Number  337  

6.  FUTURE DEVELOPMENTS  338  


6.2  Mini‐STRs  339  

6.3  Deep sequencing  339  

6.4  Alternative DNA amplification methods  340  

6.5  Mobile technologies  340  


  

  

  

322 

1. Abstract 

  

This review focuses on the discovery, recovery, identification and individualization of 

biological material important in forensic investigations. Several techniques will be 

discussed that are currently applicable in the field of forensic biology. Furthermore 

new techniques and developments that might become applicable to the field of 

forensic biology in the near future will be discussed. The aim of this review is to give 

a summary of new developments since Fourney et al 1 published a review in Interpol’s 

Forensic Science Review (2007). The first chapter will deal with the discovery and 

recovery of biological trace material at the crime scene. The second chapter 

concerns individualization of the traces and focuses mainly on the biological 

approach to identifying body fluids, where ribonucleic acid (RNA) will play an 

increasingly prominent role. The last chapter deals with the individualization of crime 

scene samples based on the deoxyribonucleic acid (DNA) content. The principal 

topics this review covers are divided into: Trace evidence discovery and recovery; 

Identification of blood, semen, and other body fluids; Individualization based on the 

genetic fingerprint, epigenetic markers, short tandem repeats (STRs), single 

nucleotide polymorphisms (SNPs), and present challenges (e.g. Quality Control). 

Furthermore some techniques will be presented that could become valuable in future 

forensic biological examinations. As in any review a selection had to be made on 

many of the scientific topics papers that are relevant to the forensic scientist. 

Therefore not all topics that are relevant to forensic biology have been dealt with. 

Topics that relevant to forensic biology and are more than worth to be covered in 

future reviews include Ancestry Informative Markers (AIMs), mtDNA analysis, and 

nonhuman biological traces (e.g. microbial forensics). 

2. Biological trace evidence 

2.1 Introduction 

  

Crime scene investigation is the first and a crucial step in forensic casework. The 

time an investigating team can spend at a crime scene is limited due to 

environmental factors, for example after a drive-by shooting a road must be screened 

for evidence as soon as possible so it can be re-opened for the public in order to 

prevent a chaos. After the road is re-opened any remaining traces will be destroyed, 

thus an investigating team has only one opportunity to collect the evidence. It is 

therefore essential that techniques are being developed that can give immediate 

results to aid the investigating team on how they should proceed. Such techniques 

should for example aim at the detection and identification of biological traces. 

2.2 Biological traces 

  

Presumably when we think of biological traces found at a crime scene, blood comes 

to mind first. Besides blood, there are many more biological traces that are 

interesting for forensic examinations (e.g. skin cells, semen, and hairs). This review 

focuses on the identification of five different body fluids being: blood, semen, saliva, 

vaginal fluid, and urine. Because body fluids all serve a different function and can be 

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present in different criminal cases they are of importance for forensic examinations. 

For example blood is responsible for the transportation of oxygen throughout the 

body, when someone is stabbed blood can be spilled from the body. The blood could 

be transferred to the perpetrator’s clothing during a struggle, if the perpetrator throws 

his clothing in a nearby trashcan the DNA from the blood can reveal that this clothing 

contains blood from the victim. If then skin cells can be retrieved from the inside of 

the clothing that have been shed by the perpetrator the DNA from these cells can be 

used for the individualization process of a possible suspect. 

2.3 Detection of traces 

  

Some biological traces are readily visible, while others can only be detected with 

special equipment. The techniques that are commonly used in today's practice are 

presumptive in nature, meaning that they only give an indication on the nature of the 

trace but are not confirmatory. Although such tests can give a valuable investigative 

lead, there is always a need for immediate confirmation to positively identify the 

nature of the stain. Preferably tests should be non-destructive to assure that the 

sample is preserved. For example, the more DNA can be collected from a sample the 

more easy it is to obtain full typing results using conventional DNA typing methods. 

Virkler and Lednev 2 propose to use fluorescence spectroscopy and/or Raman 

spectroscopy for the detection of traces. 

2.3.1 Fluorescence spectroscopy 

In fluorescence spectroscopy, radiation (e.g. ultraviolet light) is absorbed by an 

analyte (e.g. nucleic acids, proteins, and lipids). The analyte will in return emit light 

with a specific longer wavelength. Because many different analytes are found in body 

fluids they all have a certain emission spectrum (Figure 1). If this were to be 

developed in a handheld instrument it would become possible to quickly scan a large 

region of stained material. The technique is very sensitive and moreover such an 

instrument would be able to identify different body fluids. 

Figure 1. The fluorescence emission spectra of semen (—), skin oil (‐ ͵͵ ‐ ͵͵ ), blood (‐ ‐ ‐), saliva (‐ ͵ ‐ ͵ ), and urine (———)  

excited at 260 nm (by Virkler and Lednev 2 )  

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2.3.2 Raman spectroscopy 

Raman spectroscopy uses near infrared light for excitation. Although this technique 

requires sample preparation (the required amount can be as low as picograms or 

femtoliters), the sample (e.g. swab) can still be used for laboratory DNA testing. The 

result of Raman spectroscopy is a Raman spectrum (Figure 2), which provides a 

vibrational signature of the sampled material. A portable Raman spectrometer is 

already available 3 . 

Figure 2. Raman spectra, with 785‐nm excitation (by Virkler and Lednev 2 )  

2.3.3 XRF 

Besides fluorescence spectroscopy and Raman spectroscopy, X-ray fluorescence 

(XRF) is a presumptive test worth mentioning. Originally XRF was designed for 

elemental analysis during planetary exploration, though it can also be used to detect 

blood due to the presence of iron in haemoglobin, and semen due to the presence of 

zinc. This technique is particularly valuable because it is non-destructive compared to 

other screening techniques. 

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2.4 Recovery of traces 

  

After being detected biological traces have to be recovered for subsequent laboratory 

analysis. The three most commonly used methods for sampling exhibits are: cutting, 

swabbing, and taping. The use of adhesive tape has several advantages compared 

to the other collection methods: it has the ability to collect shed epithelial (skin) cells 

from crime scene items such as the inside of a glove, combined with a reduced 

collection of PCR inhibitors (e.g. clothing dyes); furthermore the inside and outside 

can be sampled separately, thus avoiding surface-to-surface contamination and 

consequently reducing the risk of obtaining mixtures with possibly irrelevant DNA 

sources. Barash et al 4 have examined four different types of tapes, and determined 

which type of tape can be used for collecting biological material while in addition also 

assures that subsequent DNA analysis can be performed. They have been able to 

identify adhesive tapes that meet these requirements. Although the use of adhesive 

tapes for the collection of biological material has its benefits, special care should be 

taken when using them on the outside of a case item. Toothman et al 5 describe that 

the presence of dust in the evidentiary item can be a significant contamination source 

in forensic investigations, due to the fact that the human DNA component in the dust 

can be sufficient to interfere with the STR analysis. On the other hand dust can also 

be a useful source of biological evidence. For example to obtain DNA profiles from 

inhabitants within a defined location 5 . 

3. Evidence identification 


  

After being detected and recovered, biological traces have to be identified. This 

chapter will focus on the biological approach to identify five different body fluids: 

blood, semen, saliva, vaginal fluid, and urine. As mentioned before the human body 

contains different body fluids, which all serve a different function. As mentioned 

before blood is responsible for the transportation of oxygen throughout the body. The 

review article by Virkler and Lednev 2 provides the framework for this section. They 

discuss the current techniques as well as the new emerging techniques. This review 

will focus on confirmatory tests that can be applied at the crime scene. 

A distinction can be made between different body fluids based on their component 

makeup. This principle is the general idea for the identification of body fluids. In Table 

1 a summary is given of some fluid specific components. Even though some 

components are fluid specific, there are others which are not. For example prostate 

specific antigen (PSA) is a protein produced by the cells of the prostate gland, and 

therefore is present in semen, but can also be found in for example breast milk and 

vaginal fluid, be it in a much lower concentration 6 . Proving its presence of PSA is 

therefore presumptive, whilst proving the presence in combination with other 

components could be confirmatory. 

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Table 1. Composition of body fluids (by Virkler and Lednev 2 ) 

Blood Semen 

-Acid 

Saliva Vaginal fluid Urine Sweat 

-Hemoglobin phosphatase 

-Prostate-specific 

-Amylase -Acid phosphatase -Urea -Urea 

-Fibrinogen antigen -Lysozyme -Lactic acid -Creatinine -Lactic acid 

-Erythrocytes -Spermatozoa -Mucin 

-Buccal epithelial 

-Citric acid -Uric acid -Chloride 

-Albumin -Choline 

cells -Urea -Chlorine 

-Tamm- 

Horsfall 

-Sodium 

-Glucose -Spermine -Thiocyanate -Vaginal peptidase glycoprotein -Potassium 

-Glycogenated 

- 

-Immunoglobulins -Semenogelin -Potassium epithelial cells 

Immunoglobulins 

3.2 Blood 

-Zinc -Bicarbonate -Acetic acid 

-Citric acid -Phosphorus -Pyridine 

-Lactic acid -Glucose 

- 

-Squalene 

-Fructose Immunoglobulins -Immunoglobulins 

-Urea 

-Ascorbic acid 

-Immunoglobulins 

3.2.1 Current confirmatory techniques 

In today's forensic practice potential bloodstains that have been discovered at the 

scene of the crime are in first instance screened with presumptive tests like the 

Kastle-Meyer (KM). This presumptive blood tests meets practically all requirements 

that a diagnostic test should have: sensitive, high specificity, quick, simple, and safe. 

The tests that can be subsequently used for the confirmatory identification of blood 

are amongst others: the microscope test, spectroscopic tests, and immunological 

tests 2 . 

The microscope test involves the counting of visualized blood cells, this can be 

expanded by the use of a scanning electron microscope (SEM) which is able to show 

the morphology of the cells present in the stain. An energy dispersive X-ray (EDX) 

can also be used to show the elemental composition of the stain. 

Spectroscopic tests, such as UV-vis(ible) absorption, determine if haemoglobin and 

its derivatives are present in the stain, which can confirm the presence of blood in a 

stain 2 . Spectroscopic tests can yield a positive result when blood is present in very 

dilute amounts. Nevertheless this technique also has its limitations and 

disadvantages, for example results can be interfered by: the exposure to sunlight, 

heating, and rust. 

A disadvantage of both tests mentioned above is the lack of species specificity. A 

positive result means that we are dealing with blood, which can be from a human 

being but also from any other animals. To show if the blood in a stain is human For 

example the Rapid Stain Identification test (RSID) can be used. The test is specific 

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for human blood by using antibodies that are specific for the human specific and 

blood specific protein glycophorin A 7 . 

3.2.2 Emerging and future techniques 

New techniques which are developed for the detection of blood are aiming on the 

identification of the body fluid, and are species specific as well. Most of the new 

techniques deal with RNA analysis. The DNA from a cell’s nucleus is transcribed into 

messenger RNA (mRNA), which in turn is translated into proteins. The proteins in a 

cell determine its function, and therefore the protein makeup together with the RNAs 

in a cell can disclose the tissue or fluid type we are dealing with. Because of the rapid 

developments in DNA technology, the step towards the use of RNA technology is 

straightforward. In 2004, Alvarez et al 8 revealed a method to isolate RNA and DNA 

simultaneously from the same stain extract. This makes it possible to yield 

information about the nature of the stain using RNA analysis, while DNA analysis of 

the same extract can be used to individualize the sample. Juusola and Ballantyne 9 in 

2005 proposed a multiplex reverse transcription-polymerase chain reaction (RT-PCR) 

to identify different body fluids (blood, saliva, semen, and vaginal fluid). The RNA 

tests to identify aimed at the genes for beta spectrin (SPTB) and porphobilinogendeaminase 

(PBGD). Haas et al 10 found that positive results could be obtained from 

stains that were up to 15 months old. Menstrual blood was detected by the presence 

of the matrix metalloproteinase-7 (MMP-7) and MMP-11, Both proteinases are absent 

from circulatory blood and menstrual-free vaginal swabs 11,12 . Zubakov et al 13 in 2008, 

analyzed gene expression in aged bloodstains in order to find stable RNA markers, 

which are highly expressed in blood, and have low expression in other fluids. They 

concluded with nine possible markers 13 (CASP1, AMICA1, C1QR1, ALOX5APm 

AQP9, C5R1, NCF2, MNDA, and ARHGAP26). Although all of the markers were able 

to differentiate blood from semen samples, none of them could differentiate blood 

from vaginal fluid because of the complex nature of vaginal secretions. In 2009, 

Zubakov et al 14 used the nine markers mentioned above on old bloodstains, and 

proved their effectiveness in freshly dried blood samples, and stains up to at least 16 

years old. Because the age of crime scene sample can vary to a great extent it needs 

to be proven that mRNAs intended for forensic use can be amplified successfully 

under a wide range of sample conditions including aged samples. 

Besides mRNA, microRNA (miRNA) can also be used for body fluid identification. 

MiRNAs are non-protein coding molecules with important regulatory functions; many 

have tissue specific expression patterns 15 . They are on average 22 nucleotides long 

and have a stem-loop secondary structure, making them less prone to the 

degradation process compared to mRNA. In 2009 Hanson et al 16 were the first to use 

miRNAs for forensic body fluid identifications. Although no truly body fluid specific 

candidates were identified (in the sense of being present in one body fluid and 

completely absent in the other body fluids). They concluded that miR-451 and 

miR-16 could be used to differentiate blood from semen, saliva, and vaginal fluid 16 . 

cDNA can be produced from all RNA species within a sample, including mRNAs and 

other small noncoding RNAs 16 . One advantage is, that when the miRNAs are 

insufficient to identify the body fluid, mRNAs can be used from the same RT reaction 

to help identify the body fluid. Furthermore the production of cDNA from total RNA 

uses less genetic material from evidentiary samples, which is crucial when there is 

little genetic material present. In this case 1ng of total RNA from blood, saliva, vaginal 

fluid, and menstrual blood extracts and 5ng of total RNA from semen extracts were 

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used in the RT reaction. The expression of miRNAs was subsequently examined 

using a SYBR Green real time PCR assay. Unfortunately this procedure cannot be 

multiplexed and has to be performed in a separate reaction for every miRNA 16 . 

Nonetheless attempts could be made to develop multiplex reactions using multiple 

dyes. Zubakov et al 15 in 2010 reported three miRNAs (miR-20a, miR-106a, and 

miR-185) that have high expression levels in circulatory blood relative to semen, 

saliva and vaginal fluid. They used the TaqMan MicroRNA Reverse Transcription 

assay, which is species specific, however there are miRNAs which do appear in other 

species (e.g. non-human primates). Of the three miRNAs mentioned above only 

miR-106a is human specific. 

3.3 Semen 

3.3.1 Current confirmatory techniques 

The tests that can be used for the identification of semen are the microscope test, 

immunological tests, and electrophoretic methods 2 . 

The microscope test can be used for the detection of semen. The microscopic 

detection of sperm cells gives a direct identification of semen. Another advantage of 

the microscope test is the possibility to show the presence of semen in a mixture with 

blood and vaginal fluid. Unfortunately not all males have sperm cells in their 

ejaculate, meaning that the absence of sperm cells does not mean there is no 

semen. The absence of sperm cells can be explained by azoospermicity of the donor 

due to natural causes or by vasectomy. To be able to identify semen without sperm 

cells other techniques have been developed. An example is the use of prostate 

specific antigen (PSA), as mentioned in the introduction, for the identification of 

semen. Semen of azoospermic males will still contain this protein which is present in 

the seminal plasma 6 . PSA can be detected in semen which is 10 6 times diluted 2 . Due 

to its extreme sensitivity other body fluids (e.g. breast milk and vaginal fluid) that also 

contain PSA but in much lower concentrations, can produce a false positive result. In 

addition PSA can give a false positive reaction on male urine 17 . 

Semen can also be detected using a lateral flow immunochromatographic strip test. 

This test is a part of the Rapid Stain Identification test (RSID TM ) and is developed to 

detect the presence of semenogelin. The detection limit of this test is 2.5nL of human 

semen, and takes about 10 minutes to give a result. While it gives no positive result 

when tested on blood, saliva, vaginal fluid, and urine. 

3.3.2 Emerging and future techniques 

Like with blood, the newly developed techniques are aiming at identification and are 

species specific as well. Most of these new techniques deal with RNA. As mentioned 

before Alvarez et al 8 revealed a method to isolate RNA and DNA simultaneously from 

the same bloodstain extract. This method can also be applied to semen samples. 

Naturally the RT-PCR method described by Juusola and Ballantyne in 2005 also 

applies to semen. Juusola and Ballantyne 9 used the semen specific markers PRM1 

and protamine 2 (PRM2). Less than 200 pg of input RNA was needed for a positive 

result with both genes. The specificity of these genes for semen has been shown, 

because they were not detected in saliva, blood, menstrual blood, and vaginal fluid 9 . 

Accordingly Fleming and Harbison 11 mention that PRM1 and PRM2 have been found 

in ejaculated spermatozoa and dried semen stains, while being absent in blood, 

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sperm-free vaginal swabs and saliva. Sakurada et al 18 used the PRM2 gene together 

with the semenogelin 1 (SEMG1) to show the presence of semen in a stain. Both 

genes are being expressed in semen, however also 2 out of 5 urine samples showed 

some expression, but this is presumably caused by semen contamination of the 

urine. 

The same as with blood microRNAs can be used for semen identification. Hanson et 

al 16 reported two miRNAs (miR-135b and miR-10b) that can be used to differentiate 

semen from blood, saliva, and vaginal fluid. Zubakov et al 15 reported five miRNAs 

(miR-135a, miR-10a, miR-507, miR-891a, and miR-943) that have high expression 

levels in semen relative to blood, saliva and vaginal fluid. Of these miRNAs only 

miR-891a is human specific. 

3.4 Other body fluids 

  

Although other body fluids such as saliva, urine and vaginal fluid are encountered 

less frequent than blood and semen in forensic practice the unambiguous 

identification of these fluids can be of great value in certain forensic cases. The 

traditional method for identifying saliva is based on the detection of the alpha 

amylase enzyme. The classic test for urine is based on the detection of urea an 

organic compound that concentrates in urine. Both the amaylse test and the urea test 

are relatively insensitive and not specific for the body fluid of interest. At present 

there is no specific and sensitive presumptive test to screen for the presence of 

vaginal secretions. The coming availability of specific RNA tests for the identification 

of these body fluids will therefore be of great value for the forensic identification of 

these body fluids. 

3.4.1 Saliva 

The genes that can be used to identify saliva are statherin (STATH) and HTN3 12,18 . 

Both genes were not detected in blood, semen, and vaginal fluid. The HTN1 gene 

was also detected by the HTN3 primer and therefore showed some false positive 

results in semen 2 . Zubakov et al 13 concluded with five possible markers for saliva 13 

(SPRR3, SPRR1A, KRT4, KRT6A, and KRT13) SPRR3 and SPRR1A show no 

detectable expression in semen, and the remaining three (KRT4, KRT6A, and 

KRT13) show vast over-expression in saliva compared to semen. However the 

markers could not differentiate saliva from vaginal fluid, due to the complex nature of 

vaginal fluid. In addition, the markers could at least be identified in 6 months old 

stains. Furthermore Zubakov et al 13 remarked that tissue identification should not 

only be based on the presence of certain body fluid markers, but also on the absence 

of the other body fluid markers. In 2009 Hanson et al 16 were the first to use miRNAs 

for forensic body fluid identifications. For the differentiation of saliva from blood, 

semen, and vaginal fluid they concluded that miR205 and miR658 could be used 16 . 

Besides a molecular biology approach saliva can be identified using a lateral flow 

immunochromatographic strip test. This test is a part of the Rapid Stain Identification 

test (RSID TM ) and is developed to detect the presence of salivary α-amylase. The 

detection limit of this test is 1µL of human saliva, and takes about 10 minutes to give 

a result. 

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3.4.2 Vaginal fluid 

The genes that can be used to identify vaginal fluid are human beta-defensin 1 

(HBD-1) and mucin 4 (MUC4). A positive result is also observed in menstrual blood, 

but not in circulatory blood, semen, and saliva. Menstrual blood can be differentiated 

from circulatory blood and menstrual free vaginal swabs by using the gene matrix 

metalloproteinase 11 (MMP11) 11 . For the differentiation of vaginal fluid from blood, 

semen, and saliva miR124a and miR372 can be used 16 , although the miR372 

concentration is only slightly higher in vaginal fluid compared to other body fluids. 

3.4.3 Urine 

Until now no genes have been described that are truly specific to urine. Nevertheless 

Nakazono et al 19 described a method to show the presence of human urine. They 

identified human urine by detecting five 17-ketosteroid conjugates with electrospray 

ionization liquid chromatography mass spectrometry (ESI-LC-MS). At the same time 

they used diafiltration to purify DNA from the sample and full DNA profiles could be 

successfully obtained using capillary electrophoresis. Remarkable however, is that 

male urine contains less DNA than female urine. 

Urine can also be identified using the Rapid Stain Identification test (RSID TM ) and is 

developed to detect the presence of the urine specific Tamm-Horsfall protein (TPH). 

The detection limit of this test is 5µL of human urine, and takes about 15 minutes to 

give a result. 

3.4.4 Sweat 

Until now no genes have been described that are truly specific to sweat. 

Nevertheless one study showed that it is possible to identify sweat using the G-81 

monoclonal antibody and an ELISA technique 2 . 

3.5 RNA as a tool for evidence identification 

  

Most emerging techniques used for body fluid identification, discussed above, are 

dealing with RNA. Unlike DNA, the transcript, RNA has not been extensively used in 

forensic science. The potential of RNA so far has been underestimated. RNA was 

notorious for its allegedly rapid post-mortem and in vitro decay, making it less useful 

in forensic investigations. However nowadays new RNA technologies are developed 

and several studies show that RNA can be detected after a long time 14,20 . 

Nevertheless the environmental conditions play a major role. The review by Bauer 21 

in 2007, described three examinations RNA could be used for. First diagnosis of the 

cause of death, second determination of the age of wounds and injuries, and last 

identification of body fluids. The main problem with these examinations are the 

unpredictable conditions forensic stains are exposed to. In order for RNA to be useful 

in crime scene investigations it is necessary to know more about the degradation 

process, furthermore it is important to know what storage conditions are preferred for 

crime scene samples. Both aspects will be discussed below for mRNA and 

microRNA. 

3.5.1 mRNA 

Setzer et al 20 performed a study to gain insight into the process of body fluid specific 

RNA degradation. The mRNA genes used for this study were: PBGD and SPTB for 

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lood; HTN3 and STATH for saliva; PRM1 and PRM2 for semen; MUC4 for vaginal 

fluid; and S15 as housekeeping gene. Samples that were exposed to the natural 

outside environment exhibited dramatically reduced recovery compared to room 

temperature and light exposed samples. UV- and luminescent light did not have an 

impact on the recovery values of RNA from blood and saliva samples. Semen 

samples exhibited increased RNA recovery. It is speculated that the light treatment 

enhances RNA isolation from spermatozoa by facilitating the disruption of cellular 

integrity. Vaginal samples however exhibited a significant reduction in RNA recovery 

upon treatment with UV, but not luminescent light. This might be caused by 

germicidal actions, as a vaginal swab is expected to contain microbial communities 

from the female reproductive tract. More research can be expected, unravelling the 

molecular basis of mRNA degradation, including whether the 5’ end, the 3’ poly-Atail, 

or the intervening sequences are specifically degraded or whether nucleolytic 

damage is randomly distributed. 

3.5.2 MicroRNA 

As discussed above, RNA degradation is promoted by environmental factors (e.g. UV 

light, moisture, and heat) 15 . This degradation will negatively affect the detection of 

RNA. To reduce this negative effect, miRNAs can be used instead of mRNA, 

because of their shorter amplicon length 16 . Zubakov et al 15 , published that blood and 

semen samples aged for 1 year under lab conditions (relatively constant humidity 

and ambient temperature, no UV exposure, dust free, etc.) still contained a 

detectable amount of miRNAs. It seemed that the aging had no negative effect on the 

absolute level of miRNAs that could be detected. Furthermore the effect of 

environmental factors on miRNA still needs to be investigated. Although it can be 

expected that miRNA degradation is influenced by the same environmental factors as 

mRNA. 

4. Evidence individualization 


  

After samples are collected and have shown to be of value for further examination 

(by e.g. identity testing) the individualization process begins. Individualization is used 

to prove with a certain amount of confidence that the evidence originates from the 

suspect or another individual, thereby linking the biological trace evidence to its 

donor. 

4.2 DNA typing 

  

Base sequence variations in the human genome can be used for DNA typing, which 

is also known as DNA identification analysis, identity testing, DNA testing, genetic 

fingerprinting, genotyping, or DNA profiling. DNA as a tool for the individualization of 

biological samples was introduced by Sir Alec Jeffreys in 1985 22 . Every human 

(except identical twins), each possesses a unique genotype, furthermore the DNA of 

an individual is identical whether it is extracted from buccal cells, white blood cells, or 

spermatozoa (note that each spermatozoa contain only half of the DNA due to 

meiosis). These principles of individual uniqueness and identical DNA structure within 

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all tissues of the same body provide the basis for DNA profiling 23 . DNA typing has 

completely pushed away conventional genetic typing methods such as blood group 

typing and analysis of polymorphic enzymes. Forensic DNA typing can be used to 

identify a suspect and to show someone's innocence. DNA typing methods can also 

be used to prove kinship in civil cases and for the identification of individuals after a 

mass disaster (flight accidents). 

4.3 Epigenetics 

  

Besides variations in the genotype, there are the so called epigenetic variations. 

Epigenetics is to some degree responsible for phenotypic alterations 24 . Most 

phenotypic alterations are caused by the genetics of an individual, but how does 

epigenetics work when we know that the base sequence of the DNA is not altered? 

The expression of certain genes may be increased by or inhibited through 

biochemical modifications of the DNA and histones 25 , the proteins around which the 

DNA coils that are crucial for DNA compacting into chromatin. Neither the gene 

structure nor the nucleotide sequence is modified. The process of biochemical DNA 

modification involves methylation of DNA molecules in many sites and acetylation, 

phosphorylation, methylation and ubiquitination of histones 24 . Such epigenetic 

modifications, so-called epigenetic markers, appear progressively throughout an 

individual’s life and with increasing frequency as the individual ages. They are the 

unexpected explanation to the phenotypic variations observed during aging. They 

probably also explain some of the phenotypic differences observed among 

monozygotic twins. A case study by Bach 24 revealed that about one third of identical 

twin pairs show differences in DNA methylation patterns. Furthermore nothing 

indicates today that epigenetic markers are transmitted beyond a limited number of 

generations. 

4.4 Short Tandem Repeats 

  

Of all the DNA in a cell approximately 2% is coding for heritable traits, while the other 

98% is ‘non coding’ 26 . Genetic mapping and sequencing studies have led to the 

discovery of highly variable regions 27 in the non-coding part of DNA between different 

individuals. These hypervariable regions, often termed minisatellites 22 and 

microsatellites (or short tandem repeats - STRs), are generically known as VNTRs 

(Variable Number Tandem Repeats) and are found at many sites throughout the 

genome. Each VNTR contains a defined sequence of nucleotides which is repeated 

a number of times in a tandem fashion, for example TCAT TCAT TCAT TCAT, the 

greater the number of repeats, the longer the VNTR. The number of tandem repeats 

in any particular minisatellite or microsatellite varies from one person to another. This 

is exploited in identifying individuals on the basis of their DNA profile. The size of the 

fragments identified will depend on the number of repeats that each microsatellite 

contains, and the pattern (a series of peaks 28 ) obtained after capillary electrophoresis 

(CE) is characteristic of the profiled individual 29 , except for identical twins, although 

they might be distinguished when examining epigenetic markers 24 . 

Fourney, DesRoches and Buckle 1 discussed the developments in STR technology 

until 2007 and mentioned the usage of Y-STRs when examining genetic genealogy. 

In the following three years new developments occurred and some will be discussed 

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here. Y-STRs are obtained by profiling the male sex chromosome, unfortunately 

when female offspring are under investigation this genetic genealogy on the basis of 

Y-STR profiling cannot be performed (note that only males have a Y chromosome). 

However the X-chromosome men transmit to their daughters is the single Xchromosome 

they received from their mother. Bobillo et al 30 in 2008 tried to show the 

applicability of X-STRs in two cases which were unable to be solved by conventional 

typing approaches. They used ten X-STR loci (DXS8378, DXS9898, DXS7133, 

GATA31E08, GATA172D05, DXS7423, DXS6809, DXS7132, DXS9902, and 

DXS6789) and were able to obtain results that did/didn’t support kinship claims. 

Aquino et al 31 in 2009 compared an autosomal STR kit (using 15 STRs), with those 

supplemented by an X-chromosome STR hexaplex (DXS7133, DXS7424, DXS8378, 

DXS6807, DXS7423, and DXS8377) in kinship cases where putative paternal 

grandmothers or half sisters were genotyped in the absence of the alleged father. In 

this study, the likelihood ratios (LR) calculated with the autosomal STR kit were 

around a 100. While in combination with the X-STR hexaplex the LR increased 6,500 

times 31 , proving its potential in complex genetic genealogy cases. Of course 

supplemental autosomal STR loci can also be used to improve resolving complex 

kinship cases 32 . 

4.5 Single Nucleotide Polymorphisms 

  

Another genetic marker that can be used for identifying individuals besides the STR 

is the single nucleotide polymorphism (SNP). Approximately 85% of human genomic 

variation is based on SNPs, meaning there is an abundance available for identity 

testing. Nevertheless not all SNPs are the same, some can be used for identity 

testing while others are useful AIMs 33 . Identity testing requires SNPs to have high 

heterozygosity and low population heterogeneity 34 . As the name already infers SNPs 

have only one nucleotide that needs to be detected, therefore the amplicon length is 

reduced and is in the range of 50bp. The short amplicon length is important when 

degraded material is under investigation, where the higher molecular weight STR 

markers can to amplify, while the SNP markers may still provide valuable information. 

In addition SNPs have a low mutation rate, which makes them useful for paternity 

testing and complex kinship cases. Furthermore stutter peaks do not occur in SNP 

analysis 33 . However due to the fact that most SNPs are bi-allelic 35 they are less 

informative for identity testing than an STR locus. On the other hand non-binary 

SNPs (e.g. tri-allelic or tetra-allelic) are suited for the detection of a second DNA 

source in a sample, a DNA mixture in a ratio up to 1:8 can be recognized by the 

presence of three alleles on one locus, unexpected peak height ratios, and uncalled 

peaks above the detection threshold 33 . To reach a discrimination power that is equal 

to 12-16 STRs a large battery of approximately 45-65 bi-allelic SNPs is required. A 

new alternative to PCR has been developed to detect SNPs 36 . This technique, rolling 

circle amplification (RCA), will be discussed below in the future development section. 

Until now little attempts have been made to develop forensic SNP multiplexes. 

Nevertheless SNPs could become implemented universally 34 . 

4.6 Polymerase Chain Reaction 

  

Isolated DNA can be present in limited quantities in evidential material. In order to 

assure enough material is available for analysis the polymerase chain reaction (PCR) 

  334 

can be used. PCR is a cyclic process in which target DNA, for example an STR, is 

being duplicated in vitro. The PCR process was developed in 1985 37 . PCR is widely 

used in DNA profiling in circumstances where there are only minute quantities of 

starting material. One cycle of the PCR process consists of denaturation of the DNA, 

binding of the primers, and elongation of the new DNA strands 38 . Because this cycle 

can be repeated multiple times the concentration of DNA will increase exponentially. 

The amount of amplicons (target DNA sequences) after a certain amount of PCR 

cycles can be defined by 2 n – (2n+2), where n is the number of cycles. In forensic 

cases somewhere between 28 and 34 cycles are used 28 . When the amplification 

process is finished the PCR mixture will almost exclusively consist of amplicons, 

therefore only the amplicons will be visible during further analysis. Every PCR 

process requires at least two primers, one on each site of the target DNA. By 

selecting suitable primers, highly variable regions or STRs can be amplified from very 

small amounts of DNA. However the complementary primer sequence has to be very 

specific, to ensure that only the target DNA is amplified. Furthermore PCR primers for 

STR loci have to bind as close as possible to the tandem repeat to decrease the 

amplicon length, because a shorter template STR has more chance of surviving 

degradation, and therefore increasing DNA profiling success. Current PCR 

techniques are able to amplify multiple STRs at the same time (this process is called 

multiplexing), meaning that only a small amount of DNA is needed and consumed for 

DNA typing (for more information on multiplexing readers should refer to Budowle 

and van Daal 34 ). 

In forensic investigations typically at least ten different loci are examined 26,28 , 

including amelogenin for sex determination. The loci that are examined are the same 

in every sample. All loci have their own specific code, for example D21S11, FGA and 

TH01. Every locus (singular of loci) has its own set of DNA-features, if all the features 

of the different loci are found it is said to be a full DNA-profile. The information from a 

DNA-profile is used to decide, whether any two samples of DNA are from the same 

individual or not 29 . The usage of the same standard set of loci by different countries 

provides the ability to compare a DNA-profile with different DNA databases on an 

international level. The Interpol standard set of loci (ISSOL) currently preferred is 

given in Table 2. Recently a DNA match in an unsolved murder, which occurred in 

the Netherlands in 1996, has led to the arrest of a German man. The DNA profiles 

were exchanged on the basis of the Treaty of Prüm. The Netherlands is affiliated with 

the Treaty of Prüm, which has now been transposed into European Union legislation. 

Because of this, DNA profiles of different countries can be compared on a daily basis. 

The Netherlands can compare DNA profiles with Germany, Austria, Spain, 

Luxembourg, Slovenia, Finland, and France. This has already led to a total of 1103 

international matches, of which 629 occurred in 2009. 

  335 

Table 2. The Interpol Standard Set of Loci (ISSOL), identical with the European Standard 

Set (ESS), is recommended by the European Network of Forensic Science Institutes 

(ENFSI). 

Supplier 

Locus NGM TM 

Applied-Biosystems Promega 

SGM Plus Profiler Plus 

D21S11 X X 

Profiler 

X 

Plus 

FGA X X X 

VWA X X X 

THO1 X X 

D3S1358 X X X 

D8S1179 X X X 

D18S51 X X X 

Amelogenin X X X 

X 

Identifiler 

Power-Plex 16 & 17 

X 

Power-Plex 

X 

16 & 17 

X X X 

X X 

X X 

X X 

X X 

X X 

X X 

D1S1656* X X 

X 

D2S441* X 

D10S1248* X 

D12S391* X X 

D22S1045* X X 

Additional Loci 

TPOX CSF1P0 D13S317 D7S820 D5S818 D16S53 

9 

  336  

X 

X 

D2S1338 D19S433 

Penta D Penta E FES F13A1 F13B SE33 CD4 GABA 

* Following new recommendations by the European Network of Forensic Science Institutes the 

ISSOL was expanded in 2010 from the existing seven to twelve markers 39 . 

5. Present challenges 

5.1 Quality Control 

  

A consistent high quality of forensic science disciplines is of vital importance for 

criminal proceedings. Judges need to work with the evidence that is presented to 

them: it has to be valid and reliable which makes the implementation of 

comprehensive quality control procedures necessary. The report by the ‘Committee 

on Identifying the Needs of the Forensic Sciences Community’ 40 shows that the 

quality between different forensic science disciplines varies greatly mainly due to 

absence of solid scientific research (proving validity of the different techniques),

quantifiable measures of reliability and accuracy, quantifiable measures of 

uncertainty in the conclusions, and the lack of research on observer bias and sources 

of human error. DNA analysis however has proven to give consistent results, with a 

high degree of certainty, sufficient to demonstrate the connection between evidence 

and a specific individual or source. In this respect the stochastic effects observed 

during PCR and especially Low Copy Number typing (LCN) can complicate the 

procedure. To make no mistakes, or have a zero error rate is practically impossible. 

Therefore it should be a good scientific practice to show the limits of a specific 

method and to address for example the impact of potential bias. Thompson 41 gives 

an example of a blind procedure, preventing observer bias, in forensic DNA profile 

interpretations. He first argues that “the widespread failure to adopt blind procedures 

for making DNA interpretations sets the stage for target shifting”. In which target 

shifting means that examiners change their interpretation on the basis of the 

available reference profiles from suspects which they know are likely to match. 

Thompson 41 proposes to prevent this by sequential unmasking, in which the features 

of the contested material are observed first. After this the examiner has to point out 

which DNA profiles could contribute to this material. Only after that he can continue 

his investigation, comparing the reference samples with the contested material. 

As the DNA technology is advancing, the statistical part becomes more and more 

important. Hedman et al 42 proposed a statistical model for unbiased quality control of 

forensic DNA profiles (e.g. capillary electrophoresis electropherograms). The 

measure proposed is the forensic DNA profile index (FI). Where the total peak height 

of the capillary electrophoresis electropherograms (e.g. the sum of the heights of the 

observed STR peaks given in relative fluorescent units (rfu)) together with the mean 

local balance (e.g. the mean of intra-locus balances or discrepancies between peak 

heights within a heterozygous STR marker), and the Shannon entropy (e.g. 

discrepancies, or inter-locus balance, between the sum of the peak heights between 

the markers) 42 are combined. The FI can be used to ensure the overall quality of 

DNA profiling within a laboratory. 

5.2 Low Copy Number 

  

In order to analyze samples that contain less than 200 pg of template DNA 28,43 (in 

literature referred to as low template DNA (LTDNA) or touch DNA) LCN can be used. 

There are several variants of the LCN technique, the most basic (the variant with 34 

PCR cycles, a replication strategy with two or more replications, and a validated 

interpretation methodology) is used in the Netherlands, the UK and in New 

Zealand 44 . LCN uses STRs to produce a DNA profile. In comparison to the 

conventional STR methods applied in normal casework LCN uses methodologies to 

increase DNA detection sensitivity. For example modifications to the PCR (e.g. other 

primers) and/or post-PCR manipulations 34,28,43,45 . 

The increase in detection sensitivity gives a concomitant increased risk of 

contamination. For example more sensitivity of detecting DNA, enhances the chance 

to find not only the DNA profile of the perpetrator, but also the DNA profile of other 

persons 46 (whose cells were present on e.g. the victim due to casual contact 28 ). 

Furthermore stochastic effects are also more likely to occur. Stochastic effects are 

heterozygote peak imbalance, allele or locus drop-out, and stutter (for more 

  337 

information on stochastic effects readers should refer to Butler and Hill 47 ). To prevent 

stochastic effects several measures can be taken: (i) prevent contamination of the 

samples 28 (e.g. by instructing and training crime scene investigators, police, and 

forensic laboratory personnel), (ii) replicating the LCN analyses (reproducibility of 

alleles in replicates is the basis for reliable results), and (iii) improving the recovery, 

extraction, and PCR 34,48,49,50 (e.g. increasing the efficiency of recovery and yield from 

a collection device could make LCN unnecessary, because sufficient material is 

obtained to successfully undertake conventional STR analysis). Furthermore it is 

important to decide, before collecting samples, if samples should be considered for 

subsequent LCN typing in order to prevent contamination. Police forces should be 

made aware of the limitations the LCN procedure has. Of course contamination must 

also be prevented during ‘normal’ sample collection. Caddy et al 28 report that until 

then (2008) there is no standardization in interpretation amongst all LCN providers. 

Furthermore the success rate of LCN typing is not yet available. The main reason for 

this lies in a lack of agreement on what constitutes a success 28 (is it a full DNA 

profile? or is it a particular number of ‘clear’ peaks in the capillary electrophoresis 

electropherogram?). Caddy et al further mention that as a matter of best scientific 

practice a DNA quantification step should be implemented before the actual (LCN) 

DNA analyses. The quantification step can be used to reveal the presence of 

possible PCR inhibitors, and can be used to reduce the chances of generating over 

amplification of the PCR products. The importance of validation of LCN typing 

methods has been made clear in a number of papers 28,43,44,47,51,52 including 

interpretation of LCN mixed profiles 44,47,52 . Caddy et al propose that LCN-produced 

DNA profiles should always be reported to the jury with the caveats that: the nature of 

the starting material maybe unknown; that the time of DNA transfer cannot be 

inferred; and that the opportunity for secondary transfer is increased in comparison to 

conventional DNA profiling 28 . 

6. Future developments 


  

The use of DNA in forensic science began 20-25 years ago. Since then various 

techniques have been developed and implemented. It can be assumed that no 

dramatic changes in current techniques will be developed, but that merely 

refinements will be made, although forensic scientists should be aware that they 

have to do anything in their power to embrace new technologies that will benefit 

society for example by their ability to analyze more challenging samples 34 . An 

example of a refinement is in the forensic DNA databases that currently contain DNA 

profiles with STRs (there are >8 million reference profiles in the CODIS database and 

>90 thousand reference profiles in the Dutch database). Implementing new STRs (as 

in the ISSOL) and removing the older ones ensures that databases can be used on 

an international level while it infers that a part of the existing databases are no longer 

useable (losing potentially valuable information). Although valuable information might 

be ‘lost’ some changes have to be made, because most countries currently have 

their own specific set of STRs making it difficult or virtually impossible to search for 

DNA profiles in databases from other countries. Furthermore if more loci can be 

compared, the strength of the evidence in case of a match will increase. 

  338 

6.2 Mini-STRs 

  

Improving the success rate of DNA profiling with the use of STRs requires more 

specific STR designs. Reducing the amplicon length for example, as discussed in the 

SNP section, will make the STRs less prone to the degradation process (this 

improvement relies on the same principles as the mRNA to miRNA conversion). 

Amplicons in the range of 71-250bp are called mini-STRs 34 . Because mini-STRs will 

overlap in size there are more fluorescent dyes required to distinguish them. Mulero 

et al 53 describe the conversion of eight STRs into mini-STRs and Gill 54 et al describe 

the acceptance of three mini-STRs into the European standard Interpol loci. 

6.3 Deep sequencing 

  

Determining the nucleotide sequence in a DNA fragment is called sequencing. The 

first DNA sequencing experiments were conducted in 1977 by Maxam and Gilbert 

and shortly after by Sanger 37 . Since then many modifications and improvements 

have been made to the widely adopted Sanger method. New sequencing 

technologies have the ability to generate huge numbers of sequencing reads per 

instrument run. Such sequencing is often referred to as deep sequencing 55 . Deep 

sequencing methods are 454-, Solexa-, SOLID-, Polonator-, and 

HeliScope-sequencing 56 . 454 Pyrosequencing, is based on the detection of released 

pyrophosphate (PPi) during DNA synthesis. A cascade of enzymatic reactions 

generates visible light with an intensity that is proportional to the number of 

incorporated nucleotides 57 , for example adenine (A) is complementary to thymine (T), 

thus when adenosine becomes incorporated it binds to thymidine, when the template 

sequence is TTA or TTTA the adenosine will be incorporated two or three times 

respectively. The amount of light produced is detected by, for example, a photodiode. 

Furthermore because the added nucleotide is known the sequence of the template 

DNA (e.g. A-G-C-T-A-G-C-T) can be determined after multiple cycles 56 . It is however 

essential that all free nucleotides are removed at the end of every incorporation step 

by either washing or enzymatic degradation 57 . 

Pyrosequencing is suitable for STR analysis 58 , moreover the markers with simple 

repeats consisting of only two different types of nucleotides (e.g. A and G in Penta E) 

are easier because they do not require the other nucleotide types. Pyrosequencing is 

currently the fastest method for sequencing a PCR product 57 and it is user-friendly 59 . 

The potential of pyrosequencing lies in its ability to be automated. Furthermore the 

technique dispenses with the need of labelled primers, labelled nucleotides, and gel- 

or capillary-electrophoresis 57 . The cost per genotype for Pyrosequencing is 1.5 USD 

(including PCR) compared to 1.9 USD per genotype using the Power Plex 16 

(Promega) and 1.5 USD per genotype for the AmpFLSTR Identifiler kit (Applied 

Biosystems) 58 . Nevertheless pyrosequencing also has its limitations and drawbacks. 

The multiplexing capability is limited 58 , e.g. multiplex sequencing can only be 

performed if all markers have different repeat structures 59 , therefore more research is 

needed to improve this capability. Furthermore pyrosequencing uses high cycle 

numbers of PCR, which can make the technology more prone to stochastic effects. 

Despite the increase in PCR cycles, no allele drop-in has been observed in a study 

by Divne et al 58 . 

  339 

6.4 Alternative DNA amplification methods 

  

Currently the PCR amplification method is used in forensic DNA analysis. However 

methods based on PCR are considered to be complex because it requires 

sophisticated and expensive equipment and skilled operators to perform the assay 36 . 

PCR may introduce false positive results from nonspecific amplification or allelic 

drop-in (due to slippage, following a random genetic drift model 43 ) because of the 

thermal cycling steps 36 . Furthermore PCR does not have true multiplexing 

properties 60 . Therefore the forensic community has to be aware of alternative DNA 

amplification methods that are being developed. For example whole genome 

amplification (WGA) can be used to amplify all the DNA in a sample 34 . An alternative 

to PCR is rolling circle amplification (RCA). Hatch et al 61 proposed to use RCA in 

order to identify single nucleotide polymorphisms (SNPs). RCA provides highly 

specific DNA sequence recognition and intense signal amplification from small 

starting probes, and can be used to perform parallel screening of DNA for SNPs. 

Recently Su et al 36 described the use of RCA in combination with magnetic beads 

and electrochemiluminescence to detect point mutations (e.g. SNPs). They 

demonstrated that RCA could be used to discriminate between two alleles and can 

be applicable for the detection of a variety of biorelated markers. 

6.5 Mobile technologies 

  

The above mentioned future developments are noteworthy, nevertheless there is one 

development expected to revolutionize current DNA casework: the application of 

mobile DNA profiling technologies. Such an instrument is described by Hopwood et 

al 62 , which uses an integrated microfluidic system for the analysis of multiplex STR 

DNA profiles from reference buccal samples. Until now routine analysis of DNA 

reference samples (e.g. buccal swabs) have to undergo laboratory testing, which 

means that the samples have to be transported from the police force to a forensic 

laboratory. To assure this transportation process is efficient police forces store the 

samples until they have enough samples to dispatch. This step is time consuming 

therefore an advantage of a portable instrument would be the ability to process 

reference samples in for example a police custody suite, assuring immediate analysis 

of samples. The development of such mobile technologies has been discussed for 

many years. Integrated microfluidic systems, also called micro total analysis systems 

(microTAS) have demonstrated to be successful modules for DNA extraction, PCR 

amplification, and capillary electrophoresis (CE), offering a tremendous potential for 

miniaturizing and integrating DNA analysis in forensic STR analysis. However some 

improvements are required if the systems are to be used for probative means, 

nevertheless they can be used as a tool to provide some probable cause in the 

apprehension of potential perpetrators whose profiles have been previously entered 

into a DNA database 63 . The biggest challenge of microTAS is the extraction of DNA 

from a crude cell lysate and processing it into a full DNA profile without operator 

intervention (which would make it relatively simple to use). To ensure that a full DNA 

profile is obtained a number of variables were identified as critical: sample collection, 

DNA extraction, PCR efficiency, PCR recovery, and robustness of the CE detection 

instrumentation 62 . The major cause of variation in the system’s output is the input, for 

different samples taken from different individuals have different quantities of cells, 

releasing different quantities of DNA upon lysis. 

  340 

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forensic DNA analysis at a crime scene using a portable microchip analyzer. 

Forensic Science International 2008; 2:301-309 

  345 

Examination of 

Questioned documents/Handwriting 


MAJ Thibaud FRITZ, Mr Frederic HERRY, 

CNE Franck PARTOUCHE 

Institut de Recherche Criminelle de la Gendarmerie Nationale (IRCGN) 

Rosny Sous Bois 

FRANCE 

346

Examination of  346  

Questioned documents/Handwriting  346  


2.  SOURCES OF REFERENCES  349  

3.  STATE OF THE ART OF THE EQUIPMENT  351  

3.1  The basic optical equipment which experts in questioned documents rely  upon:  351  

3.2  The composition of inks: review of the analytical methods  352  

3.2.1  Liquid Chromatography:  352  

3.3.2  Spectroscopy:  352  

3.3.3  GC and/or MS Techniques:  353  

3.3.4  Elemental analysis:  353  

3.3.5  Miscellaneous  353  

4  BALL POINT, GEL, MARKERS (…) PEN INKS  353  

5.  INCREASE OF INKJET AND TONER PRINTING  354  

6.  TONER  354  

7.  INKJET  355  

8.  INK AGING  356  

9.  PAPER ANALYSIS  356  

10.  DETERMINATION OF WRITING OR PRINTING SEQUENCE  357  

11.  SECURITY DOCUMENTS  357  

12.  HANDWRITING  358  

13.  INDENTED IMPRESSION  359  

14.  QUALITY ASSURANCE  360  

14.  MISCELLANEOUS  361  

15.  CHALLENGES  361  

347

16.  ACKOWLEDGEMENT  361  


348


This paper aims at making an exhaustive review paper of the technical advances in the 

field of Document Examination including handwriting reported since the 15th Interpol 

Forensic Science Symposium in 2007. 

This review paper is essentially based on articles published in the major forensic or 

generalist science journals as well as presentations or posters presented at international 

forensic meetings during the period 2007 – 2010. 

The main goal is to gather every useful and relevant elements for the improvement or 

even implementation of a questioned documents forensic lab. It also aims at helping 

labs to choose some good technical orientations for new internal developments for 

coming years and to facilitate their technology intelligence. 

Due to the high number of references found (more than 200) and the variety of sources, 

it is important to underline that the scientific basis of all papers, presentations and 

posters aren’t validated by the authors of this review. 

The fields of expertise in questioned documents are various, so we decided to sort the 

bibliography regarding the main aim of each paper: handwriting, ink composition, writing 

ink analysis, toner and inkjet ink analysis, aging, printing and/or writing sequence, paper 

substrate analysis, indented impression, altered documents, security documents, Quality 

Insurance and Miscellaneous. 

The number of existing information and papers that can be useful for document 

examiners is huge. Therefore some lacks or omissions are possible. 

2. Sources of references 

The sources of references are various. The main we used were: 

a) Sources with a generalist forensic goal, but regularly publishing articles about QD 

including handwriting : 

To be mentioned: 

• Forensic Science Communications 

• Science&justice 

• Forensic Science International 

• Journal of Forensic Sciences 

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) Scientific sources hyper specialised in some professional fields but occasionally 

delivering useful papers for the QD examiners community: 

• NIP, Society for Imaging Science and Technology (IS&T), review of 

international conference on digital printing technologies, covering the widerange 

of digital printing process and ink composition. 

• Journal of the American Society for Mass Spectrometry 

• Talanta 

• Pattern Recognition 

• Journal of Chromatography 

• Journal of Pulp and Paper Sciences 

• Measurement Science and Technology 

c) Sources directly dedicated to QD examiners or specialists in security documents: 

• ASQDE, American Society of Questioned Document Examiners. 

• Journal of Forensic Document Examination 

• Infosecura (for security documents) 

• Intergraf (for security documents) 

d) Proceedings of meetings on the examination of questioned documents including 

the following: 

• The annual meeting of ASQDE 

• The triennial conference of EAFS 

• The annual conferences of AAFS 

• The annual meetings of EDEWG and ENFHEX (ENFSI) 

• International Conference on Document Analysis and Recognition (ICDAR) 

• The International Workshop on Computational Forensics (IWCF) 

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e) Finally in the specific field of technical watching, regarding printing or paper 

making processes, or handwriting it is important to underline that the participation 

to specific forums or conferences is a means of being informed of the latest 

developments impacting on our speciality and its future: 

• DRUPA 

• Paper World 

• Cartes & Identification 

• International conference on digital printing technologies (IST & NIP) 

• International Graphonomics Society (IGS) 

3. State of the art of the equipment 

UNODC is about to deliver a guide named “Guide for the development of forensic 

document examination capacity” This guide will describe the different level of 

competences of the laboratories (basics, medium and advanced) and the equipment 

requested for each level. It will be an interesting tool for each country that wants to 

implement a national structure dedicated to document examination. 

3.1 The basic optical equipment which experts in questioned documents rely 

upon: 

• Video spectral analysis stations, where the most renowned producers are 

Foster & Freeman with its range of VSCs or Projectina. Other producers 

include Regula and CTMS… 

• An Electrostatic Detection Device to reveal indented marks on paper. 

• Stereo microscopes. 

These apparatus are gradually evolving in order to face the new security measures and 

to improve comfort of use. 

For chemical analysis, the basic techniques remain TLC or HPTLC, but it is important to 

underline that they are becoming less effective because of the increase of pigmented 

inks. Therefore, it is necessary to add some other analytical techniques to cope with this 

matter. These materials are various and some bring identical or complementary results 

whether it is for ink or paper analysis. Some of them are quoted in the ink analysis and 

paper analysis parts. 

For the analysis of TLC or HPTLC plates, some publications insist on the fact that the 

discrimination results are better by observing the plates under different illuminations 

[97], [98]. 

Djozan [78] even propose a method based on image analysis of the scanned plate 

(measure of intensity profiles of RGB characteristics of the development) with an 

estimated discriminating power of 92.8%. 

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In the particular field of Handwriting, we note the emergence of some software 

dedicated to help the examiners in his writing report. One of the most distributed at this 

time is WriteOn 2. 

3.2 The composition of inks: review of the analytical methods 

The expert in documents is brought to examine, in the majority of the cases, three major 

classes of ink: 

• Inks of pen ([66] to [102]). 

• Toner ([103] to [111]). 

• Ink of inkjet printer ([112] to [119]). 

Whilst there are many publications on the forensic analysis of inks of pen inks (37 

papers, posters or presentations found), fewer have been written on the analysis of 

inkjet inks (8 papers, posters or presentations found) and toner inks (7 papers, posters 

or presentations found). However, it seems that today, it is this kind of inks that are 

regularly asked to be analysed by magistrates or investigators more than pen inks. 

To allow further analysis, the document examiners are often compelled to preserve the 

sample he has to analyse. 

Today, Raman spectroscopy ([72], [93], [108], [109], [122], [125], [128], [146], [148]), 

remains the reference process but we can see some emergent techniques able to make 

non destructive analysis applicable to inks. 

As said below, it is extremely hard to give an exhaustive list of analytical techniques that 

can be used for ink analysis. Listed below are the references appearing in the review 

grouped according to technique, the relative popularities of which are likely to be due to 

their availability in the university or lab undertaking the tests. 

3.2.1 Liquid Chromatography: 

• TLC (Thin Layer Chromatography) [74], [78], [80], [81], [85], [95], [97], [98], [119], 

[129], [140] 

• HPTLC (High Performance Thin Layer Chromatography) [80], [81], [91] 

• HPLC (High Performance Liquid Chromatography) [91], [104], [119], [134], [140], 

[141] 

3.3.2 Spectroscopy: 

• FTIR spectroscopy (Fourier Transform Infra Red) [76], [85], [89], [108], [119], 

[126], [127], [129], [144], [147], [156] 

• µFTIR [104], [128] 

• Raman spectroscopy [72], [93], [108], [109], [122], [125], [128], [146], [148] 

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3.3.3 GC and/or MS Techniques: 

• Pyrolysis / GC / MS (inkjet and toner) (Gaz Chromatography / Mass Spectra) [68], 

[104], [109], [119], [127], [138] 

• SPME / GC / MS (Solid-Phase Micro-Extraction / Gaz Chromatography / Mass 

Spectra) [153] 

• Thermal desorption GC MS or GCFID [96], [135], [133], [136], [141] 

• GC / MS (Gaz Chromatography / Mass Spectra) [95], [135], [137] 

• ICP/MS (inductively coupled plasma mass spectrometry) [152] 

• Secondary Ion Mass Spectrometry (SIMS) [65], [87], [90], [100], [112], [123] 

• LA ICP MS [91], [101], [105], [108], [157] 

• LDMS (Laser Desorption – Mass Spectra including LDI and MALDI-TOF MS) 

[83], [84], [92], [114], [116] 

• ESI MS (Electrospray Ionisation Mass Spectrometry) [86] 

• DART MS Direct Analysis in Real Time [73], [102], [118] 

• IRMS [157] 

3.3.4 Elemental analysis: 

• LIBS [101] 

• SEM (Scanning Electron Microscopy) [126], [127] 

• XRF including µXRF and EDXRF [74], [124], [125], [127], [128], [157], [197] 

3.3.5 Miscellaneous 

• Capillary Electrophoresis [67], [115] 

• X-Ray diffraction [126], [156] 

We can notice that, with the exception of now traditional Raman and IR (FT and ATR), a 

significant number of papers were focused on LDMS and SIMS. Apparently these two 

methods provide interesting results despite the cost for the latter. 

4 Ball point, gel, markers (…) pen inks 

These subjects were again widely covered with at least 34 references. 

One body of the work that merits highlighting is that of Neumann et al [80], [81], [82], 

where they developed in three stages a complete approach for objective forensic ink 

analysis by HPTLC using a probabilistic model. The first phase deals with the quality 

assurance process with the development of a standard methodology for analysing ink 

samples in a more reproducible way. The second speaks about the development, 

testing and selection of most suited mathematical algorithms for automatic comparison 

and finally the third phase underlines the operational applications and evaluations for 

identification and evidential value assessment. This approach may be used as a good 

framework to follow for each lab that wants to improve its existing methods or implement 

new ones. 

Papson et al [84] proposes LD MS as a good replacement tool for TLC to analyse both 

dyes and pigments of pen inks. 

353

Matthews et al [92], use LDI – TOFMS. The extraction of a single inked paper fibre 

seems sufficient to get clear mass spectrometric identification of the dyes. 

Dunn et al [83], propose an alternative to LD/MS to examine inks. It is Matrix-Assisted 

Laser Desorption Ionization (MALDI) MS, that permit a better discrimination because of 

its ability to identify multiply charged dyes. 

Causin et al [85] give complementary techniques they judge excellent to be 

implemented in an analytical scheme for ballpoint inks analysis. These techniques are 

UV-Vis spectrophotometry, TLC and FTIR. 

Adam [79] and Hammond [88], both propose improved optical methods. Adam applies a 

methodology based on Principal Component Analysis (PCA) to generate linear 

regression graphs of data sets obtained with a standard luminescence spectrometer. 

The graphs represent separation of the components of luminescence from the ink-line 

and the paper. 

Hammond uses LAB colour mode available in some software like Adobe Photoshop to 

differentiate writing instrument inks. The results are interesting (no false identification) 

but it is important to underline that all the tests were done on substrates with identical 

properties (from the same brand). 

5. Increase of inkjet and toner printing 

The continuing developments in the quality of impression, combined with the ever 

reducing cost of inkjet and toner printers, has allowed this technology to spread and thus 

be used in increasing numbers of homes, for any type of document, including for a 

criminal aim, so it is thus very natural to find this type of impression in our laboratories. 

Many publications describe the analytical methods able to give information on the 

composition of these inks. These techniques can vary according to whether the inks 

contain dyes or pigments. 

6. Toner 

Few publications make reference to the forensic analysis of printed toners. However, 

Konarowska et al [107] and Zlotnick et al [108] remind that the capacity for document 

examiners to discriminate between toner printings is interesting to fight against the 

forgeries. The investigations must be led as possible by minimizing the destruction of 

the questioned documents. 

In this context, Konarowska et al [107] uses a method of microscopic investigation at 

large magnifications to study the surface aspect of the printing. Moussa et al [106] 

specifies that no appreciable microscopic difference of the state of surface of a toner 

printing was shown from a same laser facsimile machine over a period of about 5 years. 

This information underline the fact that a laser printer can still be identified a long period 

of time after having printed a questioned document. 

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Zlotnick et al [108] suggests the use of Raman spectroscopy and micro Fourier 

transform infrared (FT-IR) analysis among other more destructive analytical techniques 

(LA ICP MS, PY-GC/MS and SEM), because of the inefficiency of the TLC technique. 

His results underline that the document examiners must multiply analytical methods to 

obtain a best discrimination of toners printings, when they compare toner based 

documents between them. 

Higashikawa et al [104] proposes a similar approach based on: SEM/EDX, PY/GC/MS, 

FTIR and HPLC. 

Besides, Zlotnick et al [108] and Higashikawa et al [104], uses the scanning electron 

microscopy (SEM) to obtain information on morphological observations or elemental 

analysis. More destructive methods can produce very discriminating complementary 

information. This is demonstrated by Higashikawa et al [104] who analyse photocopied 

characters by pyrolysis gas chromatrography coupled mass spectrometry (PY-GC/MS) 

to study resins, and by high performance liquid chromatography (HPLC) to examine 

organic additives. Szynkowska et al [105] use the inductively coupled plasma time-offlight 

mass spectrometry with laser ablation (LA-ICP-TOF-MS). 

In their work, document examiners must also answer the following question: Did the 

suspected laser printer print the toner-based questioned documents? To help them, 

Tanaka [111] suggests an approach based on the correlation of photoreceptor defects to 

electrophotographic documents. Indeed, the location of such defective areas on the 

photoreceptors are visualized by plotting the voltage data for the entire circumference 

and length of the photoreceptor using automated equipment for testing the electrical 

properties of photoreceptors in the electrophotographic industry. These defective areas 

are then compared to individual defects on the printed pages to determine whether a 

correlation between them is possible. 

7. Inkjet 

Szafarska et al [115] emphasise that chemical analysis of inkjet printer inks has become 

increasingly important in forensic research due to the extensive use of inkjet technology 

by counterfeiters. Therefore, the ability to differentiate between inkjet inks is of great 

interest in document examination. He suggests using the capillary electrophoresis 

technique, with simplified stages of extraction. 

For similar reasons, Donnelly et al [116] analyses inkjet inks by LDMS to determine the 

number of inks used by a printer and the chemical composition of the colorants. 

For his part, LaPorte et al [118] tested an emergent methodology for the identification of 

coloured inkjet inks: Direct Analysis in Real Time – Mass spectrometry (DART -MS). 

This analytical technique, virtually non-destructive, involves very little sample 

preparation and allows creation of a spectral searchable database. 

Boileau et al [119] are interested in the analysis of black inkjet printed inks with thin layer 

chromatography (TLC), high performance liquid chromatography (HPLC), pyrolysis gas 

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chromatography coupled with mass spectrometry (PyGC/MS) and attenuated total 

reflection fourier transform infrared spectrophotometry (ATR-FTIR). His results allowed 

the presentation of a classification procedure that can assist the forensic scientist in the 

examination, identification and discrimination of the different inkjet computer generated 

documents. 

8. Ink aging 

This subject is still difficult because it remains controversial. Indeed, it is commonly 

admitted that ink aging may be possible but necessitates knowing some prior elements 

like, initial ink composition, conditions of storage and interaction with the substrate... 

These parameters are often unknown in forensic cases. 

Like other ink analysis technique, research is still predominantly on ball point pen inks 

rather than on other pen inks or printing inks. 

Nevertheless, Andrasko [133] investigates the aging of inkjet ink and toner. Wang [134] 

is particularly interested in ink of fountain pens while Tomcik and. Laporte [137] 

approaches globally non-ballpoint inks (fountain, fiber tip, roller ball and gel). 

A good starting point for labs that are interested in undertaking research in this field is to 

study the Ezcurra et al [131] paper, which reviews the main studies related to the aging 

of ballpoint pen inks and gel pen inks presented these last decades. 

The studies on the aging of ballpoint ink are now traditionally based on the behaviour of 

the phenoxy ethanol component, as shown in Bügler et al [135] and Ezcurra et al [140] 

works. However, the other compounds can present an interest. Choi et al [142] mention 

the by-products of glycol and phenoxy-ethanol, whereas Berger-Karin [136] quotes 

polymers and Kirsch et al [141] the resins. 

For his part, Weyerman et al [132] propose a modelling of the aging of an ink, but 

underline that this model cannot be generalized for inks stored in different conditions on 

different papers. 

For these various tests, mass spectrometry remains the technique of choice for the 

study of the aging of ink. It is sometimes coupled with gas chromatography and with 

thermal desorption [135], [136], [141], or with liquid chromatography (HPLC, HPTLC) 

[134], [140], [141]. 

9. Paper analysis 

McGaw et al [151], [152] proposed a method to differentiate recycled papers by 

identifying trace element concentrations (Mg, Al, Mn, Fe, Sr, Y, Ba, Ce and Nd) by 

Inductively Coupled Plasma mass spectrometry (ICP MS). They concluded that Ba is 

the most discriminatory element. 

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Hrivnak et al [153] described a rapid and non destructive sampling technique based on 

SPME GC MS for the analysis of volatile organic compounds emitted by paper sheet. 

They identified 21 compounds and underlined the short sampling time, simplicity and 

economic aspect. 

Fritz et al [154] proposed an image analysis method dedicated to measure the fibre 

orientation for paper differentiation. Added to other non destructive method this 

technique allowed to discriminate papers hard to classify. 

Berger [155] allowed common copy paper discrimination by spatial frequency analysis of 

light transmission images acquired by flatbed scanner. 

Van Es et al [157] tested XRF, LA ICP MS and IRMS for the discrimination of document 

papers. They concluded that LA-ICP-MS had the highest overall discriminating power 

and that it is improved by combination with XRF and even better with IRMS. 

10. Determination of writing or printing sequence 

Bojko et al [144] worked on ATR-FTIR spectral imaging to determine the sequence of 

heterogeneous line intersections produced using ballpoint pens and laser printers 

obtaining a 100% success rate. They underlined that this technique was inefficient for 

ink-jet, gel pens, roller ball and felt tip pens… 

Saini et al [145] determined the sequence between gel pen and laser printer strokes by 

microscopic methods. They found that the best results were obtained by using an 

illumination angle of 15°. 

11. Security documents 

Document fraud dealing with security documents is a field of activity generally dealt with 

border police but with a wider spectrum of criminality than illegal immigration. 

A good collaboration with these services is necessary, first of all to stand aware of the 

last kind of counterfeits and forgeries and to bring them the lab capacities to identify 

criminal networks. 

Due to the huge amount of existing document fraud at a country size, a central lab 

facility is insufficient to answer to all national request of authentication. Thus the general 

activity of fight against document fraud must be founded on different levels of expertise. 

The “Guide for the development of forensic document examination capacity” of UNODC 

[221] proposes a pyramidal structure with a basic level then a specialist level and finally 

an advanced level. When the document lab belong to a police structure, it is useful that 

this lab occupies the advanced level and coordinates the overall architecture by 

exchanging information and undertaking operational and strategic profiling. It has also a 

training role to fulfil. 

Moreover it is the role of forensic document examiners to follow all new evolutions in the 

field of security documents as optical, electronics, covert and overt security. In this wide 

357

area of expertise, close links have to be assured with electronics lab, signal processing 

labs (image) and fingerprint labs. Besides, thanks to their experience of false 

documents, document examiners have an obligation of advising authorities responsible 

for choosing the security measures embedded in all new national documents. This role 

is necessary to avoid implementation of ineffective security and to reach agreed 

standards at each level of control (basic, specialist and advanced practitioners). 

Few publications and/or forums deal with security documents, with the exception of 

some specialized reviews such as Infosecura or the Journal of Documents Identity 

published by Keesing. Itergraf conferences also provide an interesting source of 

information. These reviews provide information on innovations in security and protection 

of printed personal data. Articles provide information on the introduction of new safety 

films, holograms or electronics [169-191]. 

New supports such as PET, ABS, PVC or polycarbonate instead of traditional paper 

require additional polymer knowledge to identify substrate sources. No reference was 

found dealing with this kind of analysis and/or the foreseeable results it could bring to 

forensic applications. 

12. Handwriting 

Like every triennial period, handwriting is a widely covered subject. Some of the papers 

are essentially focused on traditional expertise like those available in the Journal of 

American Society of Questioned Document Examiners. Regarding this traditional 

approach, Harrison et al [26] propose a paper describing what the bases of handwriting 

examination are, and quote different ASTM standards and papers. They all strengthen 

the validity of the expertise and support the principle of handwriting individuality. 

Found et al [23], Dyer et al [25], Bird et al [53], [54] and Dewhurst et al [55] delivered 

test results dealing with questioned signatures, essentially disguised and simulated. It 

principally shows that Forensic Handwriting Examiners (FHEs), thanks to their search 

strategy, are significantly more confident at identifying writers’ signature as either 

forgeries or the disguised writing of a specimen provider than lay people. They also 

show that it is easier for FHEs to identify writers’ genuine signatures than writers’ 

disguised signatures. They confirm the absolute necessity for FHEs to make a first 

analyse dedicated to evaluate if the signature is simulated or disguised before forming 

an opinion on authorship. 

Alewijnse et al [50] worked on the assessment of signature complexity based on 

previous work to validate and improve an assessment model and to investigate whether 

simulation quality is influenced by signature complexity. They conclude that complexity 

is not related to the simulation quality as expected, which may be because it was 

impossible to give a preliminary assessment of the forging skills of the simulators. 

Other papers dealt with the foreseeable future of handwriting analysis where 

conclusions of FHEs are likely be supported by automatic techniques, but these must 

remain a decision-making aid. 

358

These automatic methods will be based on a probabilistic approach where relevant 

criteria have to be chosen. That is the most difficult step since handwriting is an evolving 

biometry where relevant criteria may be different from one writer to another. 

Marquis et al [57] worked on the Bayesian evaluation of the shape of the handwritten 

letters “a”. The aim is to provide a quantitative support to handwriting evidence 

evaluation. They use Fourier Descriptors to characterise the shape of loops “a”. The 

findings demonstrate that the probabilistic methodology correctly supports either the 

hypothesis of authorship or the alternative hypothesis. 

Srihari et al [24] showed that an automatic handwriting verification system can be a 

good tool to demonstrate the individuality of handwriting, through research using the 

CEDAR FOX system. He demonstrated that handwriting of twins is less discriminable 

than that of non-twins (an overall error rate of 12.9% for twins and 3.7% for non-twins) 

and that error rates with identical twins are higher than with fraternal twins. This kind of 

use of automation can be considered as a step towards a more scientific approach to 

the field of handwriting examination. 

Durina et al [29], [41] conducted a research project responding to criticisms that earlier 

studies on the individuality of handwriting did not include populations from 

“homogeneous writing communities”, and relied on computer analysis of handwriting 

rather than on human examiners. The results of their study offered evidence to support 

that there is a high degree of inter-writer variation among writers, even in a population of 

writers who all were taught the same copybook style at the same elementary school by 

the same teachers about 40 years ago, and that FHE were able to extract features from 

the writing samples that enable them to attribute authorship. 

Walch et al [16], [18], [31], [36], [37], [39] made different papers and presentation related 

to a language independent system for handwriting identification named “Flash ID”. The 

process is based on writing segmentation (graphemes) that make the system language 

independent. The results are presented as excellent and the tool as a good support for 

FHEs. 

Brink et al [46] worked on the quantity of handwritten text needed for text independent 

writer verification and identification. For the best features, it appears that the minimum 

amount of text required is about 100 characters. To get this result, they used two 

handwriting datasets, Firemaker and IAM. 

Blankers et al [51] organised a competition between different automated biometric 

signature verification methods using on- and off-line skilled forgery detection. The 

results are promising, especially for the on-line process. It also shows that automated 

methods might be able to support FHEs. 

13. Indented impression 

There are generally few papers dealing with the issue of indented impression, however, 

we found 7 references. Some were about methods to improve the actual one. For 

359

example, Tanaka [158] worked on measurement on the characteristics of the voltage at 

the surface of the film covering the document. 

Yaraskavitch [159] et al used a similar approach, concluding, among others, that a 

period of charge decay before image development is not required when operating in 

optimal regime, which is in opposition with previous findings (Seward 1998). 

Nic Daéid [160] et al, worked more precisely on the ESDA (F&F) and its cascade 

developer. Their results showed that “humidification of the cascade developer doesn’t 

have advantage over storage in a normal environment”. They also found that “200 g of 

cascade developer gives good quality results for up to 30 traces”. 

Tolliver et al [163], underlined a very important matter, which is “the potential of the 

Electrostatic Detection Device (EDD) serving as a source for transfer of DNA”. Indeed, it 

is quasi impossible to prevent cross contamination when using this kind of device. It is 

really important to know about this issue in the sample analysis process especially when 

DNA research is done by external labs. 

14. Quality assurance 

We found only one reference dealing with quality assurance directly applied to 

Questioned document matters. 

This reference is from Ottiker [202] and raises the problem of the choice of reference 

standard for handwriting examination. In his precise case it is ISO/IEC 17020 and 

ISO/IEC 17025 that are discussed. He concluded stating that “when applying ISO 

17025 for comparative analysis, the expert is considered to be the measurement 

instrument and the ultimate goal is to validate the result of this instrument (the opinion 

given by the expert). Based on this statement, the method validation as required by ISO 

17025 consists in verifying the expert’s competence, which is exactly what ISO/IEC 

17020 mainly asks for competence assurance”. 

Thus, whatever the choice of the quality standard is, it is absolutely necessary for a 

Questioned Document Examination lab to follow an official standard which is often 

recommended by the international forensic networks. For example ENFSI recommends 

ISO/IEC 17025 and/or 17020. Moreover these international networks often propose 

technical standards regarding document and handwriting analysis. Thus, before, 

creating new methods, it is necessary to be informed of all existing standards and 

papers that can be a good basis to avoid bad practices and to facilitate the process of 

accreditation. An ideal situation, would be to take part in the technical work of these 

networks. 

Moreover a good way for a laboratory to demonstrate the quality of its work or even to 

prove its commitment to continuous improvement is to take part in the various available 

proficiency testing programmes like the ones from CTS and ST 2 AR (both handwriting 

and documents , EDEWG (documents) and ENFHEX (Handwriting). 

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In this part we decided to deal with two different aspects: the first one is the emergence 

of new technologies in the field of printing techniques, the second one is more linked to 

the use of new technologies to identify printers. 

Regarding the new technologies dedicated to produce documents, the striking one is the 

recent appearance of Computer To Plate (CTP) inkjet for offset press which was 

presented during the DRUPA forum of 2008. This method allows production of offset 

plates thanks to an inkjet process. At the end, the printed document looks under 

magnification like an inkjet document although it is an offset one. If this kind of novelty 

isn’t a problem for identifying false or authentic document by comparison it may be a 

great problem for identifying the technique used and the level of the fraud (especially for 

banknotes or security documents). A similar issue, is the appearance of the inkjet press 

with millions of nozzles, for the moment in white and black, but likely evolving in colour. 

The issue of authenticating precisely the origin of a document has always existed. 

Digital technologies can now allow this, by linking a document to its printer or scanner. 

Chiang, Khanna, Mikkilineni and al [211], [216] made interesting reviews of the existing 

techniques to embed information in the printing at the digital document level or at the 

printer mechanism level. Although these techniques are not widely disseminated, it is of 

use to at least know of their existence and detect their presence, since they can give 

accurate information for investigators. The forensic part of the “Non Impact Printing 

Conference” often deals with this kind of matter. It is a proactive way to know what could 

be the track and trace solutions for tomorrow. 

15. Challenges 

The document examination expert now has at his disposal an array of techniques, 

complementary but requiring widened competencies (chemistry, printing processes, 

image analysis, electronic for security documents, statistics and probabilistic…). Thus 

for a document examiner to remain efficient it is becoming necessary to move towards 

the specialists skills of data and signal processing, whilst remaining informed of the 

developments in the technologies of printing, the fast progress of which makes the work 

increasingly difficult. Possessing all these competencies in the single field of document 

examination is becoming almost impossible. So a way to allow general improvement of 

the methods is to integrate a questioned document facility inside a laboratory, where all 

the required activities are already undertaken (electronic, signal and image analysis…) 

16. Ackowledgement 

We would like to acknowledge, Raymond Marquis (UNIL), Isabelle Montani (UNIL), Wil 

Fagel (NFI), Laurent Köhler (INCC), Laetitia Heudt (ULg/INCC) and Meredith Miller 

361

(independent FDE, FBI certified and qualified) for having sent us interesting references 

we wouldn’t have found without their precious help and advice. 


Handwriting 

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Danjun Pu - Signature verification using a Bayesian approach - IWCF 2008 : 

Washington DC, USA - 2008, LNCS 5158, PP 192-203. 

[2] Katrin Franke - Stroke-morphology analysis using super-imposed writing 

movements - IWCF 2008 : Washington DC, USA - 2008, LNCS 5158, PP 204-217. 

[3] Azah Kamilah Muda, Siti Mariyam Shamsuddin and Maslina Darus - Invariants 

discretization for individuality representation in handwritten authorship - IWCF 2008 

: Washington DC, USA - 2008, LNCS 5158, PP 218-228. 

[4] Yau-sang patrick ChengA Study on the influence of wearing a glove on handwriting 

by chinese in Hong-Kong - Journal of American Society of Questioned Document 

Examiners - 2007, Volume 10, Number 1. 

[5] Céline Equey, Raymond Marquis and Williams D. Mazella - Original research 

paper influence of writing posture on the dimensions of signatures - Journal of 

American Society of Questioned Document Examiners - 2007, Volume 10, Number 

2. 

[6] Linda J. Hartand and Lamar Miller - Microsoft access in the questionned document 

laboratory - Journal of American Society of Questioned Document Examiners - 

2007, Volume 10, Number 2. 

[7] Raymond Orta M. and Magdalena Ezcurra G. - Neurosciences applied to 

handwriting examination - Journal of American Society of Questioned Document 

Examiners - 2007, Volume 10, Number 2. 

[8] Nazia Mehrban and Ian J. Turner - A comparison of the identifying features in 

original signatures and electronically scanned signatures - Journal of American 

Society of Questioned Document Examiners - 2008, Volume 11, Number 1. 

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in San Diego County - Journal of American Society of Questioned Document 

Examiners - 2008, Volume 11, Number 1. 

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Questioned Document Examiners - 2008, Volume 11, Number 1. 

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Society of Questioned Document Examiners - 2008, Volume 11, Number 1. 

[12] Tatiana Shlafer - A method of statistical evaluation of handwriting characteristics 

developed in Russia - Journal of American Society of Questioned Document 


[13] Martha A. Blake - Ranking handwriting attributes in the signatures of one person: A 

survey of forensic document examiners - Journal of American Society of 


[14] Linton Mohammed, Bryan Found and Doug Rogers - Genuine and disguised 

signatures - an empirical approach - American Academy of Forensic Sciences - 

Washington - 2008, Volume XIV, J1. 

362

[15] Derek L. Hammond - Skill based testing in forensic document examination : Is 

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independent system for handwriting identification - American Academy of Forensic 

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Birincioglu and H. Bulent Uner - The effect of writing speed on handwriting in 

turkish alphabet - American Academy of Forensic Sciences - Washington - 2008, 


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for handwriting derived biometric identification - American Academy of Forensic 


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JoAnn Buscaglia - Statistical characterization of writers for identification - 

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comparison between biometric and forensic handwriting individuality - American 

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complex handwriting comparisons - American Academy of Forensic Sciences - 


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handwritten characters loops - Forensic Science International - 2007, Volume 172, 

number 1, pp 10-16. 

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Examiners identification and elimination opinions on questioned signatures - 

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the handwriting of twins - Journal of Forensic Science - 2008, Volume 53, number 

2, pp 430-446. 

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examiner expertise for discriminating between forged and disguised signatures - 

Journal of Forensic Science - 2008, Volume 53, number 5, pp 1154-1158. 

[26] Diana Harrison - Handwriting Examination: Meeting the Challenges of Science and 

the Law - Forensic Science Communications - 2009, Volume 11, number 4. 

[27] Ahmed Al Haddad, Peter C. White and Michael D. Cole - Examination of a 

collection of arabic signatures - Journal of American Society of Questioned 

Document Examiners - 2009, Volume 12, number 1. 

[28] Abdulaziz Al-Musa Alkahtani and Andrew W. G. Platt - Relative difficulty of 

freehand simulation of four proportional elements in arabic signatures - Journal of 

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2. 

363

[29] Marie E. Durina and Michael P. Caligiuri - The determination of authorship from a 

homogenous group of writers - Journal of American Society of Questioned 


[30] James L. Hayes - Influence of age, gender and handedness in signature imitation - 

Journal of American Society of Questioned Document Examiners - 2009, Volume 

12, number 2. 

[31] M.A. WaIch, D.T. Gantz, J.J. Miller and J. Buscaglia - Testing the languageindependant 

flash ID system for handwriting identification in english and arabic - 

European Academy of Forensic Science Conference - 2009, 5TH conference 

EAFS , QD 08. 

[32] G. Schirripa Spagnolo - Color separation to facilitate handwriting examination - 


EAFS , september, QD 10 

[33] Nesma Houmani, Sonia Garcia-Salicetti and Bernadette Dorizzi, - A novel personal 

entropy measure confronted with online signature verification systems’ 

performance - Biometrics: Theory, Applications, and Systems (BTAS) - 2008, 

September, pp 1-6. 

[34] Nesma Houmani, Sonia Garcia-Salicetti and Bernadette Dorizzi - On assessing the 

robustness of pen coordinates, pen pressure and pen inclination to time variability 

with personal entropy - Biometrics: Theory, Applications, and Systems (BTAS) - 

2009, September, pp 1-7. 

[35] Sonia Garcia-Salicetti, Nesma Houmani and Bernadette Dorizzi - A novel criterion 

for writer enrolment based on a time-normalized signature sample entropy 

measure - EURASIP journal on advances in signal processing - 2009, june, vol. 

2009. 

[36] Mark A. Walch - Validation of an automated handwriting-derived biometric 

identification system (FLASH ID) on english and arabic writings - American 

Academy of Forensic Sciences - Denver - 2009, Volume XV, J1. 

[37] Mark A. Walch, Donald T. Gantz, John J. Miller and JoAnn Buscaglia - Evaluation 

of the language-independent process in the FLASH ID system for handwriting 

identification - American Academy of Forensic Sciences - Denver - 2009, Volume 

XV, J2. 

[38] Christopher P. Saunders and JoAnn Buscaglia - Handwriting individuality: 

Probability models, subsampling routines, and implications - American Academy 

of Forensic Sciences - Denver - 2009, Volume XV, J3. 

[39] Carole E. Chaski and Mark A. Walch - Validation testing for FLASH ID on the 

Chaski writer sample database - American Academy of Forensic Sciences - 

Denver - 2009, Volume XV, J4. 

[40] Yoko Seki - How does a document examiner differentiate authors? - American 


[41] Marie E. Durina - The determination of authorship from a homogenous group of 

writers - American Academy of Forensic Sciences - Seattle - 2010, Volume XVI, 

J3. 

[42] Amanda Hepler and Christopher Saunders - A comparison between different 

likelihood ratios for assessing handwriting evidence - American Academy of 

Forensic Sciences - Seattle - 2010, Volume XVI, J4. 

364

[43] Donald T. Gantz, John J. Miller, Christopher P. Saunders and JoAnn Buscaglia - 

New results for addressing the open set problem in automated handwriting 

identification - American Academy of Forensic Sciences - Seattle - 2010, Volume 

XVI, J5. 

[44] Cedric Neumann and Glenn M. Langenburg - On the conclusions of handwriting 

examination - American Academy of Forensic Sciences - Seattle - 2010, Volume 

XVI, J6. 

[45] Yoko Seki - Optimal variables for handwriting identification - American Academy of 


[46] Axel Brink, Marius Bulacu and Lambert Schomaker - How much handwritten text is 

needed for text-independent writer verification and identification - Proc. of 

Document Recognition and Retrieval – 2008. 

[47] Axel Brink, Harro van der Klauw and Lambert Schomaker - Automatic removal of 

crossed-out handwritten text and the effect on writer verification and identification - 

Proc. of Document Recognition and Retrieval – 2008. 

[48] Marius Bulacu, Axel Brink, Tijn van der Zant and Lambert Schomaker - Recognition 

of handwritten numerical fields in a large single-writer historical collection - Proc. of 

Document Recognition and Retrieval – 2009. 

[49] Abdulaziz Al-Musa Alkahtani and Andrew W.G. Platt - A statistical study of the 

relative difficulty of freehand simulation of form, proportion, and line quality in 

Arabic signatures - Science & Justice - 2010, Volume 50, number 2, pp 72-76. 

[50] Linda C. Alewijnse, C. Elisa Van Den Heuvel, Reinoud D. Stoel and Katrin Franke - 

Analysis of Signature Complexity - Proceedings of the 14th Biennial Conference of 

the International Graphonomics Society - 2009. 

[51] V.L. Blankers, C.E. van den Heuvel, K.Y. Franke and L.G. Vuurpijl - The ICDAR 

2009 Signature Verification Competition - Proceedings of the 10th International 

Conference on Document Analysis - 2009, pp 1403-1409. 

[52] Mohammed L., Found B., Caligiuri M and Rogers D. - Can dynamic features be 

used to discriminate between genuine, auto-simulated, and simulated signatures? - 

Journal of Forensic Science - 2010, in press. 

[53] Bird C., Found B., Ballantyne K. and Rogers D. - Forensic handwriting examiners' 

opinions on the process of production of disguised and simulated signatures - 

Forensic Science International - 2010, Volume 195, pp 103-107. 

[54] Bird C., Found B. and Rogers D. - Forensic handwriting examiners' skill in 

distinguishing between natural and disguised handwriting behaviours - Journal of 

Forensic Sciences - 2009, in press. 

[55] Dewhurst T., Found B. & Rogers D. - Are expert penmen better than lay people at 

producing simulations of a model signature? - Forensic Science International - 

2008, Volume 180, number 1, pp 50-53. 

[56] Silvia Bozza, Franco Taroni, Raymond Marquis and Matthieu Schmittbuhl – 

probabilistic evaluation of handwriting evidence: Likelihood ration for authorship – 

Journal of the Royal Statistical society – 2008, Volume 57, number 3, pp 329-341. 

[57] Raymond Marquis, Silvia Bozza, Matthieu Schmittbuhl and Franco Taroni – 

Bayesian evaluation of the shape of hadwritten characters “a” – International 

Graphonomics Society – Dijon – France – 2009. 

[58] Raymond Marquis, Silvia Bozza, Matthieu Schmittbuhl and Franco Taroni – 

Application of a multivariate Bayesian methodology to evaluate handwriting 

evidence – European Academy of Forensic Science Conference – 2009. 

365

Ink composition 

[59] Michael A. Hopper - Concerning the formation of chemical toners using a latex 

aggregation process. - NIP 23 : International Conference on Digital Printing 

Technologies - 2007, 2007, pp 252-255. 

[60] Ming-Huei Liu, Fang-Yi Hsu and Chen-Jen Yang-Preparation of polymer 

microspheres by phase-separation/aggregation (PSA) technique for color toner 

applications - NIP 23 : International Conference on Digital Printing Technologies - 

2007, pp 256-259. 

[61] Elizabeth Yuan and Sharon Hiergesell - Specialty low molecular weight polyolefins 

for digital printing applications - NIP 23 : International Conference on Digital 

Printing Technologies - 2007, pp 264-269. 

[62] Dinesh Tyagi - Polyester-based chemically prepared toner for high-speed digital 

production printing - NIP 23 : International Conference on Digital Printing 

Technologies - 2007, pp 270-273. 

[63] Dmitry Fomitchev, Hairuo Tu, Adam Mackay, Casey Whicher, Jincheng Xiong, 

William Williams, Joachim K. Floess and Doug West - Colloidal silica particles for 

toners: treatment chemistry - NIP 23 : International Conference on Digital Printing 

Technologies - 2007, pp 283-287. 

[64] Masanobu Kaneeda, Akira Inoue, Yuki Amano, Paul Brandl, Maria R Nargiello and 

Kai Schumacher - Novel fumed mixed silica-titania (FM-Si/Ti) core materials for a 

new class of external toner additives - NIP 23 : International Conference on Digital 

Printing Technologies - 2007, pp 288-291. 

[65] Masaya Shibatani, Tsutomu Asakawa, Toshiharu Enomae and Akira Isogai - 

Approach for detecting localization of inkjet ink components using dynamic-SIMS 

previous analysis - Colloids and Surfaces A: Physicochemical and Engineering 

Aspects - 2008, Volume 326, number 1-2, pp 61-66. 

Writing ink analysis 

[66] Shi, X.; Xu, Y.; Wang, Y.; Li, X. and Wang, J. - Analysis of extracts from black 

water-based pen ink by HPLC - Fenxi Shiyanshi - 2008, Volume 27, number 1, pp 

46-49. 

[67] Shih C., Liu, J., Chen B. and Lin C. - Separation of crystal violet dyes and its 

application to pen ink analysis using capillary zone electrophoresis (CZE) and 

micellar electrokinetic chromatography (MEKC) - J. Sep. Sci. - 2008, Volume 31, 

number 5, pp 893-897. 

[68] Zhao P., Wang Y., Zhao Y., Wang Y., Li X., Yao L. and Wang, J. - Classification of 

blue gel inks by using pyrolysis GC - Sepu - 2007, Volume 25, number 5, pp 762- 

765. 

[69] Wang Y., Wang J. and Yao L. - Differentiation of species of blue inks for roller-ball 

pens and the determination of relative age post-deposition - Sepu - 2007, Volume 

25, number 4, pp 468-472. 

[70] Albert H. Lyter - Micro-spectroscopic examination of writing inks - American 


[71] Nicole J. Deitz and Lawrence Quarino - Microscopic examination of blue gel inks 

after cellulase digestion of paper - American Academy of Forensic Sciences - 


366

[72] Irina Geiman, John R. Lombardi, Marco Leona and Thomas Kubic - Application of 

raman spectroscopy to the analysis of questioned documents - American 


[73] Roger W. Jones, Susan E. Lorge and John F. McClelland - Characterizing writing 

inks on paper using DART mass spectrometry - American Academy of Forensic 


[74] Rachel Dubin, Elizabeth Hirst and Walter F. Rowe - Analysis of writings made with 

black gel pens - American Academy of Forensic Sciences - Washington - 2008, 


[75] Derek L. Hammond - Discriminatory power of various dichroic filters in the 

differentiation of ballpoint pen inks - American Academy of Forensic Sciences - 


[76] Christie L. Mitchell and Walter F. Rowe - Analysis of dry erase markers - American 


[77] John A. Denman, Ivan M. Kempson, William M. Skinner, K Paul Kirkbride - 

Discrimination of pencil markings on paper using elemental analysis : An initial 

investigation - Forensic Science International - 2007, Volume 175, number 2-3, pp 

123-129. 

[78] Djavanshir Djozan, Thmineh Baheri, Ghader Karimian and Masomeh Shahidi - 

Forensic discrimination of blue ballpoint pen inks based on thin layer 

chromatography and image analysis - Forensic Science International - 2008, 

Volume 179, number 2-3, pp199-205. 

[79] Craig D. Adam - In situ luminescence spectroscopy with mutivariate analysis for 

the discrimination of back ballpoint pen ink-lines on paper. - Forensic Science 

International - 2008, Volume182 , number 1-3, pp 27-34. 

[80] Cedric Neuman and Pierre Margot - New perspectives in the use of ink evidence in 

forensic science : Part I. - Development of a quality assurance process for forensic 

ink analysis by HPTLC. - Forensic Science International - 2009, Volume 185, 

number 1-3, pp 29-37. 

[81] Cedric Neuman and Pierre Margot - New perspectives in the use of ink evidence in 

forensic science : Part II. - Development and testing of mathematical algorithms for 

the automatic comparison of ink samples analysed by HPTLC - Forensic Science 

International - 2009, Volume 185, number 1-3, pp 38-50. 

[82] Cedric Neumann and Pierre Margot - New perspectives in the use of ink evidence 

in forensic science: Part III: Operational applications and evaluation - Forensic 

Science International - 2009, Volume 192, number 1-3, pp 29-42. 

[83] Jamie D. Dunn and John Allison - The detection of multiply charged dyes using 

matrix-assisted laser desorption/ionization mass spectrometry for the forensic 

examination of pen ink dyes directly from paper - Journal of Forensic Science - 

2007, Volume 52, number 5, pp 1205-1212. 

[84] Kaitlin Papson, Sylwia Stachura, Luke Boralsky and John Allision - Identification of 

colorants in pidmented pen inks by laser désorption mass spectrometry - Journal of 

Forensic Science - 2008, Volume 53, number 1, pp 100-106. 

367

[85] Valerio Causin, Rosario Casamassima, Carla Marega, Pietro Maida, Sergio 

Schiavone, Antonio Marigo and Antonio Villari - The discrimination potential of 

ultraviolet-visible spectrophotometry, thin layer chromatography, and fourier 

transform infrared spectroscopy for the forensic analysis of black and blue ballpoint 

inks - Journal of Forensic Science - 2008, Volume 53, number 6, pp1468-1473. 

[86] Mary R. Williams, Chris Moody, Lei-Ann Arceneaux, Caitlin Rinke, Katie White and 

Mickael E. Sigman - Analysis of black writing ink by electrospray ionization mass 

spectrometry - Forensic Science International - 2009, Volume 191, number 1-3, pp 

97-103. 

[87] John Coumbaros, K. Paul Kirkbride, Gunter Klass and William Skinner - Application 

of time of flight secondary ion mass spectrometry to the in situ analysis of ballpoint 

pen previous inks on paper - Forensic Science International - 2009, Volume 193, 

number 1-3, pp 42-46. 

[88] Derek L. Hammond - Validation of LAB color mode as a nondestructive method to 

differenciate black ballpoint pen inks - Journal of Forensic Science - 2007, Volume 

52, number 4, pp 967-973. 

[89] Warnadi Dirwono, Jin Sook Park, M.R. Agustin-Camacho, Jiyeon Kim, Hyun-Mee 

Park, Yeonhee Lee and Kang-Bong Lee - Application of micro-attenuated total 

reflectance FTIR spectroscopy in the forensic study of questioned documents 

involving red seal inks - Forensic Science International - 2010, article in Press. 

[90] John A. Denman, William M. Skinner, K. Paul Kirkbride and Ivan M. Kempson - 

Organic and inorganic discrimination of ballpoint pen inks by ToF-SIMS and 

multivariate statistics - Appl. Surface Sci. - 2010, Volume 256, number 7, pp 2155- 

2163. 

[91] K. Garzo, P. Korodi, A. Szökéné Toth and Z. Stefãnka - Application of 

chromatographic and spectroscopic analytical methods for differenciation of blue 

ballpoint pen inks - European Academy of Forensic Science Conference - 2009, 

5TH conference EAFS , QD 11. 

[92] B. Matthews, G.S. Walker and H.J. Kobus - The analysis of ball point inks on single 

paper fibres using laser desorption ionisation time of flight mass spectrometry (LDI- 

TOFMS) - European Academy of Forensic Science Conference - 2009, 5TH 

conference EAFS , september 2009, QD 13. 

[93] G.Reed N. NicDaeid, K. Savagea, K.Faulds and C.Keeley - The analysis of blue, 

black and red gel pen inks by RAMAN spectroscopy - Preliminary findings - 


EAFS , QD 15. 

[94] J. Pukkila - Thermal characterization of metallic paints and ball points pen inks on 

paper with a multipurpose PTV injector - European Academy of Forensic Science 

Conference - 2009, 5TH conference EAFS , QD 17. 

[95] Walter F. Rowe and Stephanie Moore - Examination of blue gel inks - American 


[96] Vanessa E. Abercrombie, Stephanie M.Houlgrave, Gerald M. LaPorte and Joseph 

C. Stephens - The characterization of vehicles found in ballpoint ink cartridges of 

different composition and possible implications on ink aging examinations - 

American Academy of Forensic Sciences - Denver - 2009, Volume XV, J16. 

368

[97] Stephanie M. Houlgrave, Gerald M. LaPorte, Joseph C. Stephens, Vanessa E. 

Abercrombie and Justin L. Wilson - The use of filtered lighting and infrared 

luminescence for the evaluation of writing inks analyzed using thin layer 

chromatography - American Academy of Forensic Sciences - Denver - 2009, 

Volume XV, J18. 

[98] Cara A. Plese, Sara E. Nedley and Rebecca Schuler - Hyperspectral imaging of 

TLC plates: A novel approach to ink discrimination - American Academy of 


[99] Derek L. Hammond, Sara E. Nedley and Rebecca Schuler - Hyperspectral imaging 

vs. video spectral analysis: A continued comparison of ink discrimination 

capabilities - American Academy of Forensic Sciences - Seattle - 2010, Volume 

XVI, J17. 

[100] Albert H. Lyter and Albert Schnieders - Analysis of writing inks by time of flight 

secondary ion mass spectrometry (ToF-SIMS) - A case study - American 

Academy of Forensic Sciences - Seattle - 2010, Volume XVI, J19. 

[101] Tatiana Trejos, Kristin Beiswenger, Alyssa Kress, Richard R. Hark and Jose R. 

Almirall - Forensic analysis of inks by laser induced breakdown spectrometry 

(LIBS) and laser ablation inductively coupled plasma mass spectrometry (LAICP- 

MS) - American Academy of Forensic Sciences - Seattle - 2010, Volume XVI, 

J22. 

[102] Joseph C. Stephens, Gerald M. LaPorte and Danna Bicknell - Comparison of a 

standard set of black ballpoint inks using a direct analysis in real time mass 

spectrometer (DART-MS) - American Academy of Forensic Sciences - Denver - 

2009, Volume XV, J19. 

Toner analysis 

[103] Noriyuki Iwata, Atsuyuki Watada, Katsuhiko Tani, Yokohama, Kanagawa, Hiromi 

Ikeura-Sekiguchi Tsukuba and Ibaraki - Application of scanning transmission X-ray 

microscopy for observation of organic compounds in toner particles - NIP 23 : 

International Conference on Digital Printing Technologies - 2007, pp 248-251. 

[104] Yoshiyasu Higashikawa, Hiroaki Kanno, Tsuyoshi Kaneko and Shinichi Suzuki - 

Forensic Discrimination of Trace Fragments of Photocopied Materials - Japan 

Society of analytical chemistry - 2007, Volume 56, Number 12. 

[105] M. I. Szynkowska, K. Czerski, T. Paryjczak and A. Parczewski - Ablative analysis 

of black and colored toners using LAICP-TOF-MS for the forensic discrimination of 

photocopy and printer toners - Surface and interface analysis - 2010, Volume 42, 

number 5, pp 429-437. 

[106] Candice Moussa and Michelle Novotny - Does my output change with age? - 

Journal of American Society of Questioned Document Examiners - 2009, Volume 

12, number 1. 

[107] U. Konarowska, S. Szepanczyk and K. Markiewicz - The use of optical microscopy 

to the verification of counterfeit documents on the laser printer - European 

Academy of Forensic Science Conference - 2009, 5TH conference EAFS , QD 09. 

[108] Joel A. Zlotnick and Douglas K. Shaffer - The need for research into the analysis of 

pigmented printing inks - American Academy of Forensic Sciences - Seattle - 

2010, Volume XVI, J18. 

369

[109] L. Engisch, J. Örtegren and W. Goedl - Studies on inkjet ink with confocal raman 

microscopy - NIP 23 : International Conference on Digital Printing Technologies - 

2007, pp 340. 

[110] Williams D. Mazella and Raymond Marquis - Forensic image analysis of laserprinted 

documents - Journal of American Society of Questioned Document 

Examiners - 2007, Volume 10, Number 1, june 2007. 

[111] Tobin A. Tanaka - The correlation of photoreceptor defects to electrophotographic 

(toner based) documents - American Academy of Forensic Sciences - Washington 

- 2008, Volume XIV, J16. 

Inkjet ink analysis 

[112] Masaya Shibatani, Tsutomu Asakawa, Toshiharu Enomae and Akira Isogai - 

Approach for detecting localization of inkjet ink components using dynamic-SIMS 

analysis - Colloids and Surfaces A: Physicochemical and Engineering Aspects - 

2008, Volume 326, number 1-2, pp 61-66 

[113] Shawn N. Donnelly, Trevor P. Cornell and John Allison - Analysis of inkjet-printed 

documents I : Physical and chemical challenges - American Academy of Forensic 


[114] John Allison, Trevor P. Cornell and Shawn N. Donnelly - Analysis of inkjet-printed 

documents II : Colorant analysis by laser desorption mass spectrometry - 

American Academy of Forensic Sciences - Washington - 2008, Volume XIV, J24. 

[115] M. Szafarska, A. Solarz, R. Wietecha-Poluszny, M. Wozniakiewicz and P. 

KoscieIniak - Optimization of extraction of inkjet inks from paper for forensic 

purposes - European Academy of Forensic Science Conference - 2009, 5TH 

conference EAFS , QD 05. 

[116] Shawn Donnelly, Josette E. Marrero, Trevor Cornell, Kevin Fowler and John 

Allison - Analysis of pigmented inkjet printer inks and printed documents by laser 

desorption/mass spectrometry - Journal of Forensic Science - 2009, Volume 55, 

number 1, pp 129-135. 

[117] Rebecca L. Schuler and Cara A. Plese - A novel method for the examination and 

characterization of documents printed with inkjet printers - American Academy of 

Forensic Sciences - Denver - 2009, Volume XV, J7. 

[118] Gerald M. LaPorte, Justin L. Wilson, Stephanie M. Houlgrave and Joseph C. 

Stephens - The classification of inkjet inks using accuTOF DART (Direct Analysis 

in Real Time) mass spectrometry - American Academy of Forensic Sciences - 

Denver - 2009, Volume XV, J17. 

[119] Michelle Boileau, Peter R. De Forest and John A. Reffner - The characterization of 

black inkjet computer printer inks using chromatographic and spectrophotometric 

techniques - American Academy of Forensic Sciences - Seattle - 2010, Volume 

XVI, J20. 

Miscellaneous ink analysis 

[120] James L. Streeter, Greg P. Kettering, and Kenneth B. Zercie - Invisible ink : 

Refresher for document examiners - Journal of American Society of Questioned 

Document Examiners - 2007, Volume 10, Number 1. 

370

[121] Shimoyama, M. - IR imaging of document examinations - Nippon Sekigaisen 

Gakkaishi - 2007, Volume 16, number 1, pp 25-30. 

[122] Heng H., Ke W. and Ji K. - Application of micro-Raman to the identification of ink 

mark - Guangxue Jishu - 2007, Volume 33, number 3, pp 456-458. 

[123] Lee J., Lee C., Lee K. and Lee Y. - Use of TOF-SIMS for the analysis of redsealing 

inks on paper - Appl. Surface Sci. - 2008, Volume 255, number 4, pp 

1523-1526. 

[124] Hatzistavros V. S. and Kallithrakas-Kontos N. G. - trace element ink spiking for 

signature authentication - J. Radioanal. Nucl. Chem. - 2008, Volume 277, number 

2, pp 399-404. 

[125] Janina Zieba-Palus, Rafał Borusiewicz and Marcin Kunicki - PRAXIS — combined 

µ-Raman and µ-XRF spectrometers in the examination of forensic samples - 


[126] Anna Vila, Nuria Ferrer and Jose F. García - Colored inks analysis and 

differentiation: A first step in artistic contemporary prints discrimination - Analytica 

Chimica Acta - 2007, Volume 588, number 1, pp 96-107. 

[127] A. Gambaro, R. Ganzerla, M. Fantin, E. Cappelletto, R. Piazza and W.R.L. Cairns - 

Study of 19th century inks from archives in the Palazzo Ducale (Venice, Italy) using 

various analytical techniques - Microchemical Journal - 2009, Volume 91, number 

2, pp 202-208. 

[128] Jana Striová, Gabriele Coccolini, Sara Micheli, Cristiana Lofrumento, Monica 

Galeotti, Andrea Cagnini and Emilio Mario Castellucci - Non-destructive and noninvasive 

analyses shed light on the realization technique of ancient polychrome 

prints - Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy - 


[129] H. Koivula, J.S. Preston, P.J. Heard and M. Toivakka - Visualisation of the 

distribution of offset ink components printed onto coated paper - Colloids and 

Surfaces A: Physicochemical and Engineering Aspects - 2008, Volume 317, 

number 1-3, pp 557-567. 

[130] Diane K. Williams and Kerri L. Moloughney - Thermal imaging of obliterated writing 

- American Academy of Forensic Sciences - Denver - 2009, Volume XV, J9. 

Aging 

[131] Magdalena Ezcurra, Juan M.G. Góngora, Itxaso Maguregui and Rosa Alonso - 

Analytical methods for dating modern writing instrument inks on paper - Forensic 

Science International - 2010, Volume 197, number 1-3, pp 1-20. 

[132] Céline Weyermann and Bernhard Spengler - The potential of artificial aging for 

modelling of natural aging processes of ballpoint ink - Forensic Science 

International - 2008, Volume 180, number 1, pp 23-31. 

[133] Jan Andrasko - Some examples of applications of a microthermal desorption 

device in the forensic laboratoryForensic Science International2009,Volume 54, 

number 5, pp 1055-1058. 

[134] Xiang-feng Wang, Jing Yu, Meng-Xia Xie, Ya-Tong Yao, Jie Han - Identification 

and dating of the fountain pen ink entries on documents by ion-pairing high 

performance liquid chromatography - Forensic Science International - 2008, 

Volume 180, number 1, pp 43-49. 

371

[135] Jürgen H. Bügler, Hans Buchner and Anton Dallmayer - Age determination of 

ballpoint pen ink by thermal desorption and gas chromatography mass 

spectrometry - Journal of Forensic Science - 2008, Volume 53, number 4, pp 982- 

988. 

[136} Claudia Berger-Karin, Ursula Hendricks and Jochen Geyer-Lippmann - 

Comparison of natural and artificial aging of ball point inks - Journal of Forensic 

Science - 2008, Volume 53, number 4, pp 989-992. 

[137] Ryan C. Tomcik and Gerald M. Laporte - The analysis of non ballpoint inks using 

gas chromatography/mass spectrometry : Relevance to ink dating - American 


[138] Y. Keheyan, G. Eliazyan, P. Engel and B. Rittmeier - Py/GC/MS characterisation of 

naturally and artificially aged inks and papers - Journal of Analytical and Applied 

Pyrolysis - 2009, Volume 186, number 1, pp 192-199. 

[139] Céline Weyermann, Beatrice Schiffer and Pierre Margot - A logical framework to 

ballpoint ink dating interpretation - Science & Justice - 2008, Volume 48, number 3, 

pp 118-125. 

[140] Magdalena Ezcurra G., Itxaso Velasco, Juan M. G. Góngora, M. Itxaso Maguregui 

and Rosa M. Alonso - Analysis of bic cristal medium ballpoint pen inks - Journal of 

American Society of Questioned Document Examiners - 2009, Volume 12, number 

2. 

[141] D. Kirsch, V. Guillou, P. Seiler and F. Koehler - Different analytical methods for the 

characterization of ballpoint ink entries - European Academy of Forensic Science 

Conference - 2009, 5TH conference EAFS , QD 07. 

[142] Mi-Jung Choi, Sung-Woo Park, Yale-Shik Sun and Chang-Seong Kim - The 

quantitative analysis of ballpoint pen inks solvents on paper - American Academy 

of Forensic Sciences - Seattle - 2010, Volume XVI, J10. 

Printing and/or writing sequence 

[143] James R. Daniels - Dual mode polarizing pellicle - Journal of American Society of 


[144] Katherine Bojko, Claude Roux and Brian J. Reedy - An examination of the 

sequence of intersecting lines using attenuated total reflectance-fourier transform 

infrared spectral imaging - Journal of Forensic Science - 2008, Volume 53, number 

6, pp 1458-1467. 

[145] Komal Saini, Ridamjeet Kaur and N.C. Sood - Determining the sequence of 

intersecting gel pen and laser printed strokes - A comparative study - Science & 

Justice - 2009, Volume 49, number , pp 286-291. 

[146] Thomas J. tague Jr., Marco Leona and Peng Wang - Raman spectroscopy of 

documents - Defense and Homeland Security - 2010, Avril 2010, pp 20-25. 

[147] Tamas Gal, Agnes Karoly and Judit Sandor - The use of microscopy-FTIR-ATR 

technique to determine the sequence of crossed lines - American Academy of 


[148] Tamas Gal - Examination of fraud documents by microscopy raman spectroscopy 

method - American Academy of Forensic Sciences - Seattle - 2010, Volume XVI, 

J16. 

372

[149] Magdalena Ezcurra - Is atomic force microscope a suitable tool for studying 

crossing lines? - Journal of American Society of Questioned Document Examiners 

- 2008, Volume 11, Number 2, december 2008. 

[150] N. Nic Daéid, L. Whitehead and M. Allen - Examining the effects of paper type, pen 

type, writing pressure and angle of intersection on white and dark dominance in 

ESDA impressions of sequenced strokes - An application of the likehood ratio - 


Papers 

[151] Elizabeth A. McGaw, David W. Szymanski and Ruth Waddell Smith - 

Characterization of undigested particulate material following microwave digestion 

of recycled document papers - Forensic Science International - 2009, Volume 54, 

number 5, pp 1171-1175. 

[152] Elizabeth A. McGaw, David W. Szymanski and Ruth Waddell Smith - 

Determination of trace elemental concentrations in document papers for forensic 

comparison using inductively coupled-plasma-mass spectrometry - Forensic 

Science International - 2009, Volume 54, number 5, pp 1163-1170. 

[153] Ján Hrivňák, Peter Tölgyessy, Soňa Figedyová and Svetozár Katuščák - Solidphase 

microcolumn extraction and gas chromatography–mass spectrometry 

identification of volatile organic compounds emitted by paper - Talanta - 2009, 


[154] T. Fritz, J.F. Bloch - Use of fibres orientation measurement for paper comparison - 


EAFS , QD 03. 

[155] C.E.H Berger - Objective paper structure comparison through processing of 

transmitted light images - European Academy of Forensic Science Conference - 

2009, 5TH conference EAFS , QD 06. 

[156] V. Causin, R. Casamassima, C. Marega, G. Peluso, R. Saini, A. Mango and S. 

Schiavone - The discrimination potential of X-RAY diffraction and of infrared 

spectroscopy for the forensic analysis of paper - European Academy of Forensic 

Science Conference - 2009, 5TH conference EAFS , QD 14. 

[157] A. Van Es , J. de Koeijer and G. van der Peijl - Discrimination of document paper 

by XRF, LA–ICP–MS and IRMS using multivariatestatistical techniques - Science & 

Justice - 2009, Volume 49, number 2, pp 120-126. 

Indented impression 

[158] Tobin A. Tanaka - Voltage and indentation development - Journal of American 

Society of Questioned Document Examiners - 2007, Volume 10, Number 1. 

[159] Luke Yaraskavitch, Matthew Graydon, Tobin Tanak and Lay-Keow Ng - Controlled 

electrostatic methodology for imaging indentations in documents - Forensic 

Science International - 2008, Volume 177, number 2-3, pp 97-104. 

[160] N. Nic Daéid, K. Hayes and M. Allen - Investigations into factors affecting the 

cascade developer used in ESDA-A review - Forensic Science International - 2008, 


373

[161] Larry A. Olson - Indentations produced by the document feeder mechanisms of two 

black and white photocopiers - Journal of American Society of Questioned 


[162] A. B. Giles and Audrey Giles - Electrostatic detection apparatus enhancement 

using astronomical image stacking and processing software - Journal of American 

Society of Questioned Document Examiners - 2009, Volume 12, number 1. 

[163] Diane K. Tolliver and Carl A. Sobieralski - Awareness of the Potential of the EDD 

Serving as a Source for Transfer of DNA - Journal of American Society of 


Altered documents 

[164] Atul K. Singla, O. P. Jasuja and Sarbjit Kaur - The effect of water soaking on 

ballpoint pen writings - Journal of American Society of Questioned Document 


[165] Patrick De Smet - Reconstruction of ripped-up documents using fragment stack 

analysis procedures - Forensic Science International - 2008, Volume 176, number 

2-3, pp 124-136. 

[166] P. De Smet - Two-way interactive stack desequencing for reconstruction of rippedup 

- European Academy of Forensic Science Conference - 2009, 5TH conference 

EAFS , QD 04. 

[167] Gregg M. Mokrzycki and Gabriel D. Watts - Charred documents - American 


Security documents 

[168] Carl R. Mc Clary - Design and security features of the euro - Journal of American 

Society of Questioned Document Examiners - 2007, Volume 10, Number 1. 

[169] Helene Andersson - Forged LASER engraved documents - INTERGRAF – XXII. 

International Security Printers’ Conference – Nice – 2007. 

[170] Philip Berrie - The importance of machine readibility as a security feature now, and 

five years time. - INTERGRAF – XXII. International Security Printers’ Conference – 

Nice – 2007. 

[171] Chris Catlin - Co-processing of paper and polymer banknotes. A system approach. 

- INTERGRAF – XXII. International Security Printers’ Conference – Nice – 2007. 

[172] Bill Dorricott and Emmanuel Elgart - Innovation in stamp paper making - 

INTERGRAF – XXII. International Security Printers’ Conference – Nice – 2007. 

[173] Iain Drummond - The document's own DNA as the ultimate authenticator - 


[174] Jan-Dirk Enschedé - New ideas in improving security features in banknotes - 


[175] Martyn Fry - Stamp re-use and counterfeiting in the UK. - INTERGRAF – XXII. 


[176] Réne Michael Holly - US views on data protection and biometrics - INTERGRAF – 

XXII. International Security Printers’ Conference – Nice – 2007. 

[177] Peter Hradek - Innovative use of mobile terminals for secure documents - 


374

[178] Dr. Eckhard Koch - The digital seal for security documents - INTERGRAF – XXII. 


[179] Georgy Kornilov - Innovative security features for banknote substrates - 


[180] David Lewis - A new approach to IR-absorbers for high security printing - 


[181] Luís Machado and José Rosmaninho - The introduction of the portuguese 

biometric passport - INTERGRAF – XXII. International Security Printers’ 

Conference – Nice – 2007. 

[182] Bernd Mainzer - ID cards with high data storage capacity and high security 

features in one medium - INTERGRAF – XXII. International Security Printers’ 

Conference – Nice – 2007. 

[183] Allister McCallum - Trends in eurobanknote couterfeiting lessons for the future - 


[184] Hervé Naullet - Long-range vs short-range detection of security documents - 


[185] Pandelis Papadimitriou - New developments in liquid crystal pigment technology as 

a security system - INTERGRAF – XXII. International Security Printers’ Conference 

– Nice – 2007. 

[186] Hugues Souparis - Combination of latest optical technologies on the french epassport 

- INTERGRAF – XXII. International Security Printers’ Conference – Nice – 


[187] Charlie Stevens - Development of ICAO machine readable travel documents with 

particular emphasis on biometrics and their application to border controls - 


[188] Ewout van Haeften - Counterfeiting of eurobanknotes - INTERGRAF – XXII. 


[189] Nigel Ward - Enhancing document security trough multi-factor identity verification - 


[190] Dr. Andreas Wolf - Border security, e-passports, interoperability - INTERGRAF – 

XXII. International Security Printers’ Conference – Nice – 2007. 

[191] Jürgen Zerbes - The integrative design approach to functional banknote security 

features - INTERGRAF – XXII. International Security Printers’ Conference – Nice – 


[192] D. Joseph, S. D. Maind, A. Saxena and R. K. Choudhury - Characterization of 

offset printing ink tagged with rare-earth taggants by X-ray emission techniques - 

Int. J. PIXE - 2007, Volume 17, number 3-4, pp 183-191. 

[193] Elaine Wooton, Douglas K. Shaffer and Troy Eberhardt - Methods of assessment 

of true, simulated, and counterfeit watermarks - American Academy of Forensic 


[194] Shigeru Sugawara - Passport examination by a confocal-type laser profile 

microscope - Forensic Science International - 2008, Volume 178, number 1, pp 40- 

45. 

[195] Marcel A. Verhoff, Carsten Wtzel, Kerstin Kreutz and franck Ramsthaler - The ideal 

subject dictance for passport pictures - Forensic Science International - 2008, 

Volume 178, number 2-3, pp 153-156. 

375

[196] S. D. Maind, N. Chattopadhyay, Ch. Gandhi, S. C. Kumar and M. Sudersanan - 

Quantitative evaluation of europium in blue ballpoint pen inks/offset printing inks 

tagged with europium thenoyltrifluoroacetonate by spectrofluorometry and ICP- 

AES - Science & Justice - 2008, Volume 48, number 2, pp 61-66. 

[197] Sangita Dhara, N.L. Misra, S.D. Maind, Sanjukta A. Kumar, N. Chattopadhyay and 

S.K. Aggarwal - Forensic application of total reflection X-ray fluorescence 

spectrometry for elemental characterization of ink samples - Spectrochimica Acta 

Part B: Atomic Spectroscopy - 2010, Volume 65, number 2, pp 167-170. 

[198] Pei-Ju Chiang, George T.-C. Chiu, Edward J. Delp and Jan P. Allebach - 

Optimized Embedding and Decoding of Extrinsic Signatures for 

Electrophotographic Halftone Images - NIP 24 : International Conference on 

Digital Printing Technologies - 2008, 2008 , pp 782-786. 

[199] Eva Snirer, Margaret Kalacska and Rick B. Parent - Spectral analysis of canadian 

currency as a potential tool for counterfeit detection - American Academy of 

Forensic Sciences - Denver - 2009, Volume XV, J20. 

[200] Aravind K. Mikkilineni, George T. C. Chiu, Jan P. Allebach and Edward J. Delp - 

High-capacity data hiding in text documents - Proceedings of SPIE, the 

International Society for Optical Engineering - 2009, Volume 7254. 

[201] Utpal Garain and Biswajit Halder - Machine Authentication of Security Documents - 

Proceedings of the 2009 10th International Conference on Document Analysis and 

Recognition - 2009, pp 718-722. 

Quality assurance 

[202] T. Ottiker - Accreditation of handwriting examination under ISO/IEC 17020 or 

ISO/IEC 17025 : What differences are to be expected? - European Academy of 

Forensic Science Conference - 2009, 5TH conference EAFS , QD 16. 

Miscalleneous 

[203] Linlin Zheng Rodriguez, S. Lu Zhang Botao Shao Li-Rong Zheng - Design and 

implementation of a fully reconfigurable chipless RFID tag using inkjet printing 

technology - IEEE circuits and systems - 2008, pp 1524-1527. 

[204] Jiro Oi and Hideo Taniguchi - New erase head for RFID thermal rewritable media - 

NIP 23 : International Conference on Digital Printing Technologies - 2007, 2007, pp 

148-150. 

[205] Haley A. Elliott - Decoding the United States postal service barcodes - American 


[206] Joachim Heinzl - Ink Jets - Comprehensive Microsystems - 2008, Volume 3, pp 

335-368. 

[207] Wenjuan Cuia, Wensheng Lua, Yakun Zhang, Guanhua Lina, Tianxin Wei and 

Long Jiang - Gold nanoparticle ink suitable for electric-conductive pattern 

fabrication using in previous ink-jet printing technology - Colloids and Surfaces A: 

Physicochemical and Engineering Aspects - 2010, Volume 358, number 1-3, pp 

35-41. 

[208] Douglas K. Shaffer - Evidential value of adhesives in questioned document 

examinations - American Academy of Forensic Sciences - Seattle - 2010, Volume 

XVI, J15. 

376

[209] Yoshihisa Kitano, Toshiharu Enomae and Akira Isogai - Comparative studies of 

gloss development in electrophotography and offset printing - Journal of Imaging 

Science and Technology - 2008, Volume 52, number 1, pp 010504. 

[210] John Welch - Erasable ink; something old, something new - Science & Justice - 


[211] Pei-Ju Chiang, Nitin Khanna, Aravind K. Mikkilineni, Maria V. ortiz Segovia, 

Sungjoo Suh, Jan P. Allebach, George T.C. Chiu and Edward J. Delp - Printer et 

scanner forensics - SECE, SME, Purdue University, West Lafayette, Indiana, USA 

- 2008, LNCS 5158, PP 22-34, 2008. 

[212] S.J. Simske, J.S. Aronoff, M. Strugill, F. Collins, G. Golodetz and R. Israel - 

Security printing deterrants : A comparison of TIJ, DEP and LED printing - NIP 23 : 

International Conference on Digital Printing Technologies - 2007, pp 543-548. 

[213] O. Bulan, V. Monga, G. Sharna, and B Oztan - Data embedding in hardcopy 

images via halftone-dot orientation modulation - SPIE International Conference on 

Security, Steganoigraphy, and Watermarking of Multimedia Contents X - 2009, Vol. 

6819. 

[214] S. Suth, J.P. allebach, , G.T.-C. Chiu and E.J. Delp - Printer mechanism-level 

information embedding and extraction for halftone documents : New results - NIP 

23 : International Conference on Digital Printing Technologies - 2007, pp 549-553. 

[215] Joost Van Beusekom, Faisal Shafait and Thomas M. Breuel - Document signature 

using intrinsic features for counterfeit detection - ICWF 2008 - 2008, LNCS 5158, 

PP 47-57. 

[216] Nitin Khanna, Aravind K. Mikkilineni, George T.C. Chiu, Jan P. Allebach and 

Edward J. Delp - Survey of scanner et printer forensics - IWCF 2008 : Washington 

DC, USA - 2008, LNCS 5158, PP 22-34. 

[217] Janette Dove Guscott - Case report : Can you have the perfect training case? - 

Journal of American Society of Questioned Document Examiners - 2007, Volume 

10, Number 2. 

[218] Yoshinori Akao, Atsushi Yamamoto and Yoshiyasu Higashikawa - Estimation of 

inkjet printer spur gear teeth number from pitch data string of limited length - 

Proceedings of the 3rd International Workshop on Computational Forensics - 2009, 

Volume 5718, pp 25-32. 

[219] Christian Schulze, Marco Schreyer, Armin Stahl and Thomas M. Breuel - 

Evaluation of graylevel-features for printing technique classification in highthroughput 

document management systems - Proceedings of the 2nd international 

workshop on Computational Forensics - 2008, Volume 5158, pp 35-46. 

[220] Orhan Bulan, Junwen Mao and Gaurav Sharma - Geometric distortion signatures 

for printer identification - IEEE International Conference on Acoustics, Speech and 

Signal Processing - 2009, Volume 0, pp 1401-1404. 

[221] Guide for the development of forensic document examination capacity UNODC to 

be published on the following website 

http://www.unodc.org/unodc/en/scientists/publications.html?ref=menuside 

377

MEDIA EVIDENCE 

  378 

Audio Analysis 

Review : 2007-2010 

Carl R. Kriigel, MA 1 

Geoff Smith, M.Sc. 1 

Matthew Graves, M.F.S. 1 

Forensic Audio Examiner, Digital Evidence Branch, U.S. Army Criminal Investigation 

Laboratory 

  

  

  

    

  

  

379 



2.  AUTHENTICATION (GEOFF SMITH)  381  

3.  SPEAKER IDENTIFICATION (MATTHEW GRAVES)  383  

4.  AUDIO ENHANCEMENT, RELATED AUDIO FIELDS, LABORATORY PROCEDURES AND  

WORKING GROUPS (CARL KRIIGEL)  386  

5.  RELATED FIELDS OF AUDIO EVIDENCE  387  

6.  LABORATORY PROCEDURES  388  

7.  U.S. REPORT ON FORENSIC SCIENCE  389  

8.  WORKING GROUPS AND ORGANIZATIONS  389  

9.  ACKNOWLEDGMENT  390  

10.  REFERENCES:  390  

  

    

  

  

380 


This review focuses on Forensic Audio Analysis and the advances in technology during 

the past three years. The principal topics covered are divided into five sections: 

Authentication, Speaker Identification, Audio Enhancement, Related Fields of Audio 

Evidence, and Working Groups and Organizations. 

The selection of specific sections to be reviewed was based on a broad literature review 

of approximately 100 references which was further refined. For most of the references 

the authors reviewed the complete articles; however, the abstracts were sufficient for a 

small number of reviews. 

Over the past few years there have been several areas of substantial research and 

publication. One area of interest is the use of Electronic Network Frequency (ENF) 

analysis for authentication of audio recordings. Work has been on-going for several 

years in Europe, while in the United States, research has been started in an effort to 

implement this technique. Additionally there have been various projects dedicated to 

testing automated speaker identification systems and developing better methods for 

speaker identification as well as research projects relating to speaker identification, 

each of which help strengthen the current state of the science. There also appears to 

be an increased focus on forensic audio procedures as more laboratories have 

requirements for forensic laboratory accreditation and practitioner certification. In the 

United States there is extensive discussion in the forensic science community in 

response to the 2009 release of the U. S. National Academy of Sciences released 

report entitled “Strengthening Forensic Science in the United States: A Path Forward.” 

2. Authentication (Geoff Smith) 

  

The authentication method garnering the most attention is the use of Electric Network 

Frequency (ENF) analysis that was first proposed by Grigoras in 2003 (1). It has been 

called the “'biggest breakthrough since Watergate [1972 U.S. political scandal]” (2) in 

the field of audio authentication. In this method, the frequency changes of the mains 

hum (50 Hz or 60 Hz depending on location) in a digital recording are compared to a 

historical database of the mains frequency changes. Depending on the conditions, it is 

possible to provide details about the recording including the date and time the ENF 

corresponds to, the geographical area the ENF corresponds to, if the recording is 

original or copied, and the continuity of the recording. 

For current methodologies see Grigoras (3) and Cooper (4). These methodologies 

include ENF database collection, extraction of the ENF data from a recording, and an 

automated procedure for matching the extracted data to the historical database. ENF 

analysis has been tested in both continental Europe by Brixen (5), Michalek (6), and 

  

    

  

  

381 

Grigoras and in mainland U. K. by Cooper. Sanders (7) conducted preliminary 

research comparing ENF measurements within and between each of the three major 

U.S. power grids. He found good intra-grid consistency and no correlation between the 

grids. 

The above cited methods use a mean squared error-based procedure to match the 

recording ENF data to the historical database. Huijbregtse and Geradts (8) put forth a 

matching procedure based on the correlation coefficient. They demonstrated the 

possibility of erroneous matching due to frequency offset using the mean squared errorbased 

procedure. The correlation coefficient-based procedure provided improvement 

especially for recordings shorter than 10 minutes. 

Brixen (9) and Sanders & Popolo (10) tested the susceptibility of battery-powered 

recording devices to ENF. Both tests included various recording devices and varying 

magnetic field strength. Brixen reported that no ENF was detected on a recording 

device with an electrets microphone regardless of the strength of the magnetic field. A 

majority of the recorders in the Sanders & Popolo test found no ENF at normal or 

slightly-elevated environmental levels. From these studies it can be seen that not all 

recordings will contain usable ENF data. Both studies concluded that further tests are 

needed in this subject to understand recorder susceptibility to ENF. 

Two scientific working groups have addressed ENF analysis. The ENFSI (European 

Network of Forensic Science Institutes) which has a Working Group for Forensic 

Speech and Audio Analysis (FSAAWG) published a best practices guide (11) for the 

use of ENF analysis in authentication. The second, a U.S. group, the Scientific Working 

Group on Digital Evidence (SWGDE) released a discussion paper (12) to describe the 

potential use of ENF analysis in the United States. This paper also contains 

engineering and forensic concerns with the method that should be addressed. 

Nicolalde and Apolinario (13) presented a technique of analyzing ENF data that is 

distinct from typical ENF analysis in that the ENF phase change of a recording is 

analyzed to identify discontinuities. Also analyzed were discontinuities in spectral 

distance. Using both ENF phase change and spectral distance measurements provide 

a high probability of detecting if a portion of the speech signal has been deleted from or 

inserted into a recording. 

Outside of ENF analysis publications, there have been additional advances in digital 

audio authentication. Koenig and Lacey (14) provide a thorough 11-step protocol of 

digital audio authentication. This protocol encompasses the entire examination process 

from evidence marking through reporting. The paper complements an earlier protocol 

for analog recordings from 1990 (15) with numerous changes to account for the 

  

    

  

  

382 

differences between analog and digital media. To address the need to authenticate 

digital audio, Grigoras (16) presented statistical tools to detect traces of 

(re)compression data. The tool is used to assess compression generation and to 

discriminate between different compression algorithms. In a technique specific to MP3 

compression, Yang, Qu and Huang (17) examined the frame offsets to detect 

manipulation. MP3 audio samples are divided into frames to encode with each frame 

having its own offset after encoding. Manipulation can lead to broken frame grids. 

Detection rates using this technique were reported to be 94%. In order to detect buttsliced 

edits, Cooper (18) presented a simple signal processing method having 

significant benefits over traditional approaches. This method is based on high pass 

filtering the audio and modeling a discontinuity at higher frequencies; it is used to 

complement other techniques since no single method can determine authenticity. 

While not as frequent as with digital recordings, authentication of analog recordings is 

still necessary on occasion. Visualization of magnetic features is the most effective 

method for authentication of analog recording. Boss (19) described methods of 

visualization including using crystals with polarizing microscopes, a MOSeS (Magneto- 

Optical Sensor System) device, and a MOA-KOF (Magneto-Optical Analysis) device. 

Images of visualized recordings including start and stop marks; an example is given of 

how visualization can explain certain recording peculiarities (in this case likely due to a 

tape fold). Another discussion of magnetic visualization methods was presented by 

Broyles (20). The advantages and disadvantages of ferrofluids compared to magnetooptical 

film imaging methods are discussed. Supporting test images are included. 

Koenig, Lacey and Killion (21) described the components of a macrophotography 

system to capture images of Bitter (ferrofluid development) patterns commonly used on 

audio duplication, enhancement, authentication and voice comparison examination of 

audio tapes. 

In a technique valid to both analog and digital audio recordings, Malik and Farid (22) 

described a technique to estimate the amount of reverberation in a recording. 

Differences in estimated reverberation in a recording purported to be from a single 

environment can be used as evidence of manipulation. 

3. Speaker Identification (Matthew Graves) 

  

This review will focus on speaker identification and the developments and research 

conducted in this field between 2007 and 2010. While the overview provided here 

contains many sources for information regarding speaker identification, members of the 

Digital Evidence Branch at the United States Army Criminal Investigation Laboratory 

  

    

  

  

383 

writing this review, do not perform and are not trained in this discipline. Much of the 

work is being conducted by Michael Jessen of the Budeskkriminalamt Germany (BKA). 

The two main approaches in speaker identification by experts remain the auditoryacoustic 

method and the automatic method. There have been several projects 

dedicated to testing automated systems and developing better methods and techniques 

for conducting this type of examination. In addition, many research projects relating to 

speaker identification have occurred that may help strengthen the current state of the 

science. Although testing current methodologies, techniques, and systems remains 

important, reporting of results must be considered as well. Therefore, some work has 

been dedicated to Likelihood Ratios (LRs). 

Automated systems can be used for speaker recognition. For an overview of automatic 

speaker recognition technology, see Kinnunen and Li (23). One voice comparison 

system, the SPES (SPrecher-Erkennungs-System) which is an automatic voice 

comparison system used by the BKA and is designed for forensic examinations of 

speech recordings was tested using real case data. The results show how the system 

performance is influenced by real world recording scenarios (24). Additional works by 

Chakroborty and Saha (25), Charbuillet et al. (26), Chetouani et al.(27), Liao et al.(28), 

Lu and Dang (29), Kwon and Narayanan (30), Ramos-Castro et al.(31), Shahin (32), 

and Wu and Lin (33) provide various methods and techniques for increasing the 

effectiveness of automated systems. Finally, Ramos et al. (34) address database 

mismatch from a speech database in Spanish: Ahumada III, which was created using 

data from real casework. 

Becker et al. (35) tested a new approach combining the GMM-UBM (Gaussian Mixture 

Model – Universal Background Model) framework with “formant features which have 

shown a high discrimination ability among speakers in the acoustic-phonetic approach 

of manual speaker verification.” While this technique showed promise for future use in 

forensic cases, the authors caution that additional research is needed. 

The use of “higher-level” features in automatic systems to aid in forensic speaker 

recognition was evaluated by Shriberg and Stolcke (36). In their paper, they provide an 

overview of such systems, potential advantages, and issues when using higher-level 

features in forensic applications. They state that using these features improves 

recognition accuracy because they “add the most information to a traditional low-level 

speaker recognition system.” 

In many cases, the amount of information that is available for analysis is limited. 

Therefore, it is necessary to have a method or technique for dealing with such 

situations. Gonzalez-Dominguez et al. (37) examined the use of Factor Analysis (FA) to 

overcome limited data sets that are common in forensic applications. 

  

    

  

  

384 

Occasionally, limited information may be due to the type of device that was transmitting 

the voice, for instance VoIP (Voice over Internet Protocol) communication systems. 

Two projects by Khan et al. (38,39) explored speaker recognition from data that was 

encrypted or compressed due to a communication system design. With the growing 

popularity of VoIP communications, this research could prove to be valuable to the 

forensic community in the future. 

Disguised voices may be another obstacle that some practitioners of speaker 

identification may encounter. It is possible, that if a speaker knows he/she is being 

recorded, that a disguise may be employed to mask identity. To show how a disguised 

voice could affect an examination, Zhang and Tan (40) evaluated an automatic speaker 

recognition system against 10 types of voice disguises. 

Improvements to the underlying science and validations of current methods are of vital 

importance for any discipline. One paper, by Amino and Arai (41) discusses “speakerdependent 

characteristics of the nasals” which were found to be useful for Perceptual 

Speaker Identification (PSI). Another, by Ariyaeeinia et al. (42) tests the ability of 

speaker verification technology to differentiate between twins. Additionally, Jessen (43) 

measured the articulation rate (AR) of 100 German-speaking male subjects in an 

attempt to build on multi-speaker studies in this area of research. 

Another significant area of research possibilities comes from Nolan et al. (44) and the 

Dynamic Variability in Speech (DyViS) database compiled at the University of 

Cambridge. The goals of the project are to “test the practicality of ‘speaker-space’ for 

distinguishing members of a large population of speakers, quantify articulatory-acoustic 

dynamic features for individual speakers, test diachronic change as a source of speaker 

idiosyncrasy, and make available a speech database of Standard Southern British 

English (SSBE) for wider use by other researchers, forensic phonetic practitioners, and 

other interested persons.” 

As with any analysis, the reporting of findings remains the most critical step in the 

process, since any error here could misrepresent the data the practitioner collected. 

According to Morrison (45), the forensic comparison sciences are in the midst of a 

paradigm shift that is tending towards “quantitative data-based implementation of the 

likelihood-ratio framework with quantitative evaluation of the reliability of results.” As a 

result, Morrison et al. (46) describe two procedures that are designed to estimate the 

“precision of the output of a forensic-comparison system.” While LRs may be useful for 

reporting results, it is important not to misrepresent the weight of the evidence, as 

cautioned by Gonzalez-Rodriguez (47). 

  

    

  

  

385 

In June of 2010, Michael Jessen of the BKA presented Current developments in 

forensic speaker identification at Odyssey 2010: The Speaker and Language 

Recognition Workshop. That presentation contains a wealth of information that may be 

useful to speaker identification practitioners. 

4. Audio Enhancement, Related Audio Fields, Laboratory Procedures and 

Working Groups (Carl Kriigel) 

  

To improve the intelligibility of speech it is necessary to suppress or attenuate the 

unwanted noise that occurs in audio recordings. Ellwart and Czyzewski (48) propose 

an algorithm that is based on a combination of adaptive filtration and spectral 

subtraction to reduce noise and improve the speech quality. The method utilizes two 

signals: the sound recorded by the microphone containing the speech including noise 

and the second signal containing only the noise. Several experiments were conducted 

using microphones with different characteristics. The results indicate the method 

worked well in various environmental conditions. Bitzer and Brandt (49) investigated 

the weakness of standard noise reduction approaches and offered solutions for burst 

from GSM (Global System Mobile Communication). One method for GSM burst 

removal, suggested by Bitzer and Brandt, utilized a template signal cancellation. The 

approach is to build a template of the burst signal and apply it over all detected bursts to 

remove the GSM. Bitzer and Brandt discussed other approaches to speech 

enhancement and limitations (such as low grade microphones, A/D converters and 

compression/coding algorithms). 

There are several manufacturers of commercial noise-reduction systems that contain a 

wide range of parameters that have to be adjusted by the forensic practitioners. These 

adjustments are opinion settings based on the practitioners’ experience and training but 

may not produce optimal results. Hilkhuysen and Huckvale (50) studied the differences 

between opinion based operator settings and settings using an intelligibility model. 

The intelligibility model is founded on a speech envelope distortion measure to predict 

optimal parameter settings of commercial noise-reduction systems. Hilkhuysen and 

Huckvale’s results indicate that the intelligibility model is able to predict optimal 

parameter settings better than human operators, even though the model contains 

imperfections. 

Audio restoration software in forensic applications has raised some questions regarding 

the validity of investigative and legal recordings. Koenig and Lacey (51) tested two 

widely available software programs using both speech information and sine waves. The 

results of the restoration were compared to the original source clip recording using a 

  

    

  

  

386 

variety of methods to assess the “corrections.” The results suggest that the two 

programs tested produced few improvements in the recording quality and often 

degraded the voice and other signals. The research supported that detailed testing of 

clipped-sample restoration software is strongly recommended before being utilized in 

forensic laboratories. 

Compressed digital audio recordings are being introduced to law enforcement, 

emergency and business communities as they migrate to digital media. To gain insight 

into the optimal playback and effects of digital audio recording and playback, Koenig 

and Lacey (52) tested ten small solid-state (flash memory) audio recorders. Several 

results concerning applications in forensic examinations, including authenticity, 

enhancement, voice comparison, and signal analysis were featured. Koenig and Lacy 

encouraged more research into intelligibility and suitability of using small solid-state 

recorders under typical investigative conditions. 

5. Related Fields of Audio Evidence 

Microphones play a key role in audio recordings and their use is often overlooked, 

leaving post-processing efforts as the norm to clean up these recordings. Using 

multiple microphones is more advantageous in enhancing speech signals, thereby 

improving their intelligibility. Gunel et al. (53) focused on the performance of a closedform 

acoustic scene decomposition technique. The technique blindly decomposes the 

sound field using signals obtained from a coincided microphone array. Decomposition 

both localizes the audio objects and enhances them, thereby improving the intelligibility 

of speech signals. The results indicate this technique achieves very good performance 

in realistic forensic scenarios where the intelligibility of the speech is important. 

Comparison of sounds recorded during a phone call to an emergency 911 recording 

system can sometimes be useful to the investigation. During a homicide investigation, 

the investigating department determined that the suspect had probably hit the victim on 

the head with an aluminum baseball bat. An acoustic analysis was conducted to 

determine if the recovered bat had produced the five “ping” sounds that had occurred 

during the assault. These ping sounds were recorded by the department’s 911 system 

through an open telephone line at the scene. Marr and Koenig (54) conducted this 

unique signal analysis. Several laboratory comparisons were made between the 

submitted baseball bat, two other test bats, and the 911 recording. The process used 

narrow-band spectrum analysis. The examination results indicated the resonant 

frequency of the submitted ball bat was generally consistent with the 911 ping 

recordings. 

  

    

  

  

387 

Recordings of audio gunshots can assist in crime scene reconstruction. The sound 

information can assist in estimating the shooter’s location and verify eyewitness 

accounts. Maher and Shaw (55) conducted a study by recording a systematic set of 

rifle shots. The study summarizes the primary acoustical evidence from the recorded 

gunshots and outlines several strengths and weaknesses of gunshot analysis for 

forensic purposes. 

6. Laboratory Procedures 

Forensic Laboratory procedures and practices are becoming more important as 

agencies begin to have requirements for certification and accreditation. Practices, 

policies and organization structure discussed by How (56) provide insight into the 

United Kingdom’s largest Forensic Audio Laboratory at the Metropolitan Police Digital 

and Electronics Forensic Services , London , U.K . 

Koenig, Lacey and Killion (57) provide a protocol to improve voice intelligibility of 

investigative and forensic audio recordings. Sections include the differences between 

analog and digital recordings, appropriate laboratory space, applicable equipment and 

software, examination procedures, evidence handling and expert testimony. 

Forensic audio professionals, according to Rogers and Smith (58), are beginning to 

have overlapping roles in computer forensic applications. Defining the audio 

professional role in a digital world allows for opportunities in training and understanding 

the fundamentals of the infrastructure and computing systems that create, store and 

process the audio evidence. Audio professionals are encouraged to embrace 

technology dealing with acquisition and recovery of digital media. 

The Scientific Working Group on Digital Evidence (SWGDE) prepared SWGDE Best 

Practices for Forensic Audio (59) to provide forensic audio practitioners 

recommendations for the handling and examination of forensic audio evidence in order 

to successfully introduce audio evidence in a court of law. The document covers 

laboratory equipment considerations, collection of evidence, examination, and 

documentation. The best practice guide is useful for agencies currently conducting 

audio examinations or those planning on expanding their forensic capabilities. 

The European Network of Forensic Science Institutes (ENFSI) Forensic Speech and 

Audio Analysis Working Group produced a document for ENF (Electronic Network 

Frequency) analysis in the area of forensic authentication of digital audio and 

audio/video recordings(60). The ENFSI Forensic Speech and Audio Analysis Working 

  

    

  

  

388 

Group also produced a document for Terms of Reference for Forensic Speaker Analysis 

(61) in order to successfully present reports or testimony in a court of law. 

7. U.S. Report on Forensic Science 

In 2009 the U.S. National Academies of Sciences released a report entitled 

“Strengthening Forensic Science in the United States: A Path Forward” (62) to the 

Congress of the United States. The report contained a broad overview of the status of 

forensic science in the United States along with thirteen recommendations for 

improvement. These recommendations generated significant discussion and could 

impact the forensic audio community, if adopted. SWGDE published a position paper 

(63) on the report, responding to each of the recommendations. 

8. Working Groups and Organizations 

Forensic audio analysis is a growing community that has members in several 

international working groups: 

AAFS- American Academy of Forensic Science: Within the American Academy of 

Forensic Science is the newly formed Digital and Multimedia Sciences section that 

includes forensic audio and speech analysis. http://aafs.org/digital-multimedia-sciences 

AES- Audio Engineering Society: The Audio Engineering Society is devoted exclusively 

to audio technology. Founded in the United States in 1948, the AES has grown to 

become an international organization that unites audio engineers, creative artists, 

scientists and students worldwide by promoting advances in audio and disseminating 

new knowledge and research. http://www.aes.org/ 

ENFSISA –European Network of Forensic Science Institutes Forensic Speech and 

Audio Analysis Working Group: a European group that is focused on all aspects of 

forensic audio and speech analysis, including linguistics. 

http://www.enfsi.eu/page.php?uid=63 

NCMF-National Center for Media Forensics: a newly formed American center that is 

part of the University of Colorado which has a strong forensic audio program and is 

conducting research on ENF. http://cam.cudenver.edu/ncmf/ 

  

    

  

  

389 

SWGDE- Scientific Working Group on Digital Evidence: an American group that 

includes a forensic audio committee that has produced best practices manuals and is 

promoting research on ENF. http://www.swgde.org/ 

9. Acknowledgment 

We thank Micheal Jessen of the Bundeskriminalamt Germany, Jurrien Bijhold of the 

Netherlands Forensic Institute, Catalin Grigoras and Jeff Smith of the University of 

Colorado, Denver, National Center for Media Forensics, Ken Marr of the FBI Forensic 

Audio, Video and Image Analysis Unit, David Hallimore of the Scientific Working 

Group on Digital Evidence, and Doug Lacey of BEK TEK LLC, for assistance in 

preparation this manuscript. 

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Digital Analysis 


Paul Reedy 

Manager, Forensic Operations 

Forensic and Data Centres 

Australian Federal Police. 

Michael Holohan, 

Senior legal officer, 

Crime and misconduct commission, 

Queensland, Australia 

397


1  STATISTICAL SURVEY OF CONSUMER USAGE OF DIGITAL MEDIA  400  

2  STORAGE DEVICES AND DEVICE CONVERGENCE  400  

2.1  Data Volume  400  

2.2.  Data Speed  400  

2.3  Security and Encryption  401  

2.4  Anti forensic techniques  402  

2.5  Social networking  403  

3  LABORATORY ACCREDITATION AND STANDARDS  405  

3.1  Practitioner training  405  

3.5  Tool validation  406  

4  DIGITAL FORENSIC EXAMINATIONS  407  

4.1  Evidence triage  407  

4.2  Evidence reproducibility  407  

4.3  Operating systems  408  

4.4  Hand held devices including mobile (cell) phones  408  

4.5  Forensic tools  409  

4.6  Managing investigators and prosecutors expectations  409  

5  LEGAL ISSUES  410  

5.1  Presentation of digital evidence  410  

5.2  Limitations in the Court’s understanding of digital evidence  411  

6  FUTURE TRENDS  411  

6.1  The future of digital forensics  411  

6.2  Investigative management  411  

6.3  Evidentiary problems with emerging technologies  412  

6.4  Hand held devices  412  

6.5  Practitioner welfare  412  

398

6.6  Virtualisation  413  


399

1 Statistical survey of consumer usage of digital media 

Internet usage statistics report a growth in world internet usage of 

approximately 400% from 2000 to 2009 with the largest growth regions 

being Asia, Africa and the Middle East (1). World internet penetration rates 

report that approximately 76% of the population of North America, 60% of 

Oceania and Australia and 53% of Europe use the internet (2) . 

The growth in electronic multi media is evidenced by the growth in free 

access media sharing sites such as YouTube. On October 9, 2009, the 

third anniversary of the acquisition by Google, Chad Hurley announced in a 

blog posting that YouTube was serving "well over a billion views a day" 

worldwide (3). 

2 Storage devices and device convergence 

2.1 Data Volume 

The cost of data storage halves every 18 months (4). In 2004 the cost per 

megabyte of a hard disk was $1.15; in 2009 it was 5–6 cents. Then 250GB 

was thought to be enormous (5). Today 1500 GB disks are relatively 

inexpensive costing in the order of $100. 

Casey states that the increasing capacity of commonly used storage 

media, large amounts of data on networks, expanding variety of computing 

devices, growing case loads, and limited resources are combining to 

create a crisis for Digital Forensics Laboratories (6). Casey submits that it 

is becoming critical for digital forensic procedures to deal with the mounting 

quantities of digital evidence, and to avoid causing delays in the criminal 

justice process. Laboratories, he submits, can address these problems by 

establishing thresholds to prioritize cases and scaling the forensic 

examination process to fit the needs of the case. He states that the scale 

of the examination should be dependent upon considerations that include 

the seriousness of the offence under investigation and the type of evidence 

sought. 

Casey suggests that training investigators, forensic examiners, and 

prosecutors to recognize when a survey/triage forensic inspection or 

preliminary forensic examination are appropriate will maximize the 

investigative opportunities that digital evidence can create when it is 

available (7). In doing so, Laboratories can make more effective use of 

resources, satisfy the needs of more customers, and limit case backlogs. 

2.2. Data Speed 

Harris states that advancements in computer technology have increased 

processing speeds and storage capacity, creating the need and ability to 

store large amounts of metadata for organizing and searching information 

stored in a computer (8). 

400

2.3 Security and Encryption 

Bluetooth enabled devices have become widely available and readily 

accessible to consumers in the form of mobile phones, laptop computers 

and GPS navigation systems. Browning has described Bluetooth 

technology to be vulnerable to hacking measures (9). In the case of 

mobile phones, Bluetooth technology may provide an opportunity for 

someone to listen to phone conversations or read text messages without 

the authorized user’s knowledge. Moreover, calls or texts may be made 

without authorization (10). 

Wireless access points have also emerged rapidly to provide consumers 

with widely accessible, immediate ad in many cases free and unrestricted 

access to the internet. Inherent in this technology is the vulnerability of the 

network provider and consumer to criminal liability or violation. Network 

users may be exposed to security vulnerabilities and unwittingly place 

personal information at risk of discovery or manipulation. The network 

provider may be exposed by providing a network to be abused for criminal 

purposes. Herein lies a further challenge to the digital forensic 

investigator. Takahashia reports that network protocol analyzers, typically 

developed for network performance analyses, may provide an investigative 

tool to investigate such network criminality (11). Takahashia reports that 

some characteristics in IEEE 802. (11) Traffic particularly help identify 

network users. Like actual human fingerprints, there are also unique traffic 

characteristics for each network user. They are called network user 

fingerprints, by tracking which more than half of network users can be 

connected to their traffic even with medium access control (MAC) layer 

pseudonyms. The concept of network user fingerprints is likely to be a 

powerful tool for intrusion detection and digital forensics in the future (12.) 

Growth in the use of and sophistication of digital encryption will require law 

enforcement agencies to develop critical investigative skills. Dr Glen 

Dardick, Longwood University states that current level of sophistication and 

potential for spoliation and tampering with evidence going undiscovered is 

a cause for concern (13). Applications and systems are increasingly 

becoming more sensitive to the privacy needs of individuals. These 

systems may not necessarily be deleting evidence. They may not be 

creating evidence in the first place. Applications with the specific purpose 

of deleting evidence (e.g. Evidence Eliminator, Timestomp) have now 

become off the shelf software. Dardick argues that if the digital forensic 

expert is a finder of fact, then that expert may be finding those facts more 

difficult to discover. The role of the digital forensics expert is now much 

more than a finder of fact. The expert needs to develop further critical 

investigative skill relevant to counter anti forensics. It maybe necessary to 

locate evidence of spoliation rather than the evidence itself. A clever 

analogy presented was one involving the needle in a haystack. In a digital 

world it maybe that a copy of a haystack is used to cover the needle (e.g. 

overwriting). It is the haystack and not the needle that becomes crucial 

evidence. 

401

Commercial and open source encryption tools are becoming increasingly 

popular to protect personal information from unauthorised use. Truecrypt 

is a common free open source encryption tool. The Truecrypt website 

promotes its product as enabling the creation of a virtual encrypted disk 

within a file and mount it as a real disk. It may be used to encrypt an entire 

partition or storage device such as USB flash drive or hard drive. It may 

encrypt a partition or drive where Windows is installed. Disturbingly it 

promotes the capability to rovides plausible deniability, in case an 

adversary forces the user to reveal the password (14). Truecrypt reports 

that the software has been freely downloaded on over 14.2 million 

occasions worldwide (15). 

Encryption provides a major challenge to the digital forensic investigator 

attempting to locate evidence on a digital storage device. Fortea states 

that digital forensic experts are currently working on the technological front 

together with software vendors although cooperation is not expected in this 

regard as any public knowledge of a covert backdoor in the software will 

see its marketability rapidly decline (16). 

2.4 Anti forensic techniques 

Anti-forensics is the catch-all term now used to reference research and 

developments dedicated to the idea of manipulating data to undermine the 

reliability of digital forensic investigations (17). Behr states that information 

technology security experts throughout the world are now stating that 

digital forensics is unreliable due to the sheer volume, easy access, and 

complications anti-forensic tools present for the core objectives of digital 

forensics (18). Behr describes at least four component parts to antiforensics, 

namely evidence destruction, evidence hiding, evidence source 

elimination and evidence counterfeiting (19). He concludes that while 

attempts to employ any one of the above-referenced anti-forensic 

strategies by someone who is unfamiliar with them would likely leave a trail 

of evidence that could result in significant sanctions, a thorough antiforensic 

attack may often render data-mining through computer forensics a 

practice in futility. Equally, he states, a comprehensive attack can trick 

investigators by leaving evidentiary trails that may well lead to injustices for 

the innocent resulting in confiscation of property or liberty. 

Initial research has recently commenced into the investigation and 

identification of issues relating to the analysis of the Platter Resident 

Firmware Area (20). According to Sutherland the possibility that the Platter 

Resident Firmware Area could be manipulated and exploited to facilitate a 

form of steganography will become more readily available to enable 

information to be concealed from discovery by a digital forensic examiner 

(21). Sutherland expresses a concern that this may be exploited as a 

possible route to introduce malware into a computer system. Potential 

solutions to this risk include the standardisation of some aspects in the way 

which disk firmware is structured or the generation of a library of trusted 

and true firmware which a forensic investigator can use to compare a 

suspects disk. 

402

Kuncik states that counter forensic tools have become more common and 

more powerful. None of these methods however guarantee to eliminate all 

evidence of their use and are more likely to leave some evidence of their 

use on computers, networks and servers (22). Any attempt to destroy 

evidence indicates an intention to hide data and is likely to destroy the 

credibility of those who employ it. Unfortunately an incentive to destroy 

evidence remains due to the fact that the penalty for evidence tampering 

may not be as severe as the penalty for being caught with the original 

evidence. 

Chval states that while evidence that is obtained during a cyberinvestigation 

through the application of computer forensic techniques may 

be untainted, the reality is that anti-forensic applications are pervasive and 

becoming more accessible to the general public with each passing day, 

both in terms of their general availability and their ease of use (23). 

Attempts to employ anti-forensic strategies by someone who is unfamiliar 

with them would likely leave a trail of evidence that could result in 

significant sanctions, particularly if the evidence itself is evidence of the 

use of anti forensic technologies. In contrast, Chval states that a thorough 

anti-forensic attack may often render data-mining through computer 

forensics a practice in futility. Equally, a comprehensive attack can trick 

investigators by leaving evidentiary trails that may well lead to injustices for 

the innocent resulting in the loss of property or liberty (24). Sophisticated 

users, on the other hand, will be able to wreck havoc with little concern of 

consequence. 

Chval concludes that while many courts continue to accept computer 

forensics' tools, techniques and concomitant evidence as presumptively 

reliable, both the developments in anti-forensics and research coming out 

of related fields point to a reality where computer forensic tools and 

techniques merit far greater scrutiny (25). While many courts have not yet 

adopted standards for addressing these concerns, others are busy at work 

laying that foundation (26). 

2.5 Social networking 

During the reporting period the world has seen a growth in social 

networking web sites such as Facebook and Myspace through Web 2.0 

enabling peer to peer communication and self expression through the 

sharing of autobiographical information. 

Facebook describes its mission to give people the power to share and 

make the world more open and connected. The site boasts that millions of 

people use Facebook everyday to keep up with friends, upload an 

unlimited number of photos, share links and videos, and learn more about 

the people they meet (27). The site has over 400 Million users (28). 

MySpace is also an online community that lets users “meet their friends' 

friends” (29). The site boasts that a user can create a community on 

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MySpace and you can share photos, journals and interests with your 

growing network of mutual friends (30). 

Minotti describes social networking sites as presenting discovery problems 

because information is not physically stored on the user’s computer (31). 

Berman states that Web 2.0 applications offer a framework in which its 

users interact with and even create content themselves, rapidly sharing the 

information across users and encouraging other users to respond in kind 

(32). Another aspect that social networking sites have in common is that 

they promote very informal means of communication (33). It is for this 

reason that forensic examination often unveils truly damning email 

communications. This informality according to Berman is even truer of 

Web 2.0 applications with Blogs and journal entries on social networking 

sites often being a stream of consciousness statements (34). 

National Research Council, Strengthening Forensic Science in the United 

States, review on the forensic Sciences including the discipline of digital 

forensics 

In 2009, the United States congressionally mandated report from the 

National Research Council, Strengthening Forensic Science in the United 

States, found allegedly serious deficiencies in the nation's forensic science 

system and called for major reforms and new research (35). The Report 

concluded, inter alia, that there are serious issues regarding the capacity 

and quality of the current forensic science system. Specifically, the courts 

continue to rely on forensic evidence without fully understanding and 

addressing the limitations of different forensic science disciplines. 

Moreover, it concluded that scientific evidence tests the abilities of judges, 

lawyers, and jurors, all of whom may lack the scientific expertise to 

comprehend the evidence and evaluate it in an informed manner. 

The report’s findings are applicable to the presentation of digital forensic 

evidence in criminal proceedings internationally. The challenges to the 

digital forensic discipline nationally are not insurmountable. A concerted 

effort is required by law enforcement agencies and the administrators of 

justice in criminal proceedings nationally to enhance the probative value of 

digital evidence. 

The proliferation of digital and multimedia technologies and data storage 

have provided significant challenges to law enforcement agencies and the 

administrators of justice in criminal proceedings nationally (36). The 

Report states “Moreover, in almost every instance, scientific evidence tests 

the abilities of judges, lawyers, and jurors, all of whom may lack the 

scientific expertise to comprehend the evidence and evaluate it in an 

informed manner” (37). Moreover it states that the law’s greatest dilemma 

in its heavy reliance on forensic evidence, however, concerns the question 

of whether, and to what extent, there is science in any given “forensic 

science” discipline (38). 

404

The Report poses two very important questions that it states, should 

underlie the law’s admission of and reliance upon forensic evidence in 

criminal trials: (1) the extent to which a particular forensic discipline is 

founded on a reliable scientific methodology that gives it the capacity to 

accurately analyze evidence and report findings and (2) the extent to which 

practitioners in a particular forensic discipline rely on human interpretation 

that could be tainted by error, the threat of bias, or the absence of sound 

operational procedures and robust performance standards39. 

The Report states that digital evidence has undergone a rapid maturation 

process. This discipline did not start in forensic laboratories. Instead, 

computers taken as evidence were studied by police officers and 

detectives who had some interest or expertise in computers. Over the past 

10 years, this process has become more routine and subject to the rigors 

and expectations of other fields of forensic science. Three holdover 

challenges remain: (1) the digital evidence community does not have an 

agreed certification program or list of qualifications for digital forensic 

examiners; (2) some agencies still treat the examination of digital evidence 

as an investigative rather than a forensic activity; and (3) there is wide 

variability in and uncertainty about the education, experience, and training 

of those practicing this discipline. (40) 

3 Laboratory Accreditation and Standards 

3.1 Practitioner training 

While examining existing resources to ensure the availability of both the 

skills and equipment necessary for effective forensic science facilities, the 

United Nations continue to receive a range of requests for advice and 

assistance concerning the training of staff and the selection of appropriate 

equipment and reference material (41). The United Nations developed the 

Staff Skill Requirements and Equipment Recommendations for Forensic 

Science Laboratories Manual in response to these requests and aims to 

provide practical assistance for the establishment or upgrading of national 

forensic science infrastructure general forensic science facilities and law 

enforcement technical departments. The Digital and Multimedia Evidence 

(42) discipline for example, while gradually moving in the same direction, 

continues to be performed largely by investigative personnel or 

investigative personnel re-assigned to the forensic laboratory environment. 

Only recently have advanced education programs and certification 

programs emerged specifically for this discipline, as well as the 

development of external proficiency testing providers. The manual has 

identified an array of requisite skills commensurate with the growth in 

diversity in the digital forensic discipline. These skills include: 

• Knowledge to recognize non-traditional operating systems and 

proprietary software applications, password protected files, and 

encryption 

405

• Knowledge of how data is stored on various media types and 

deleted data recovery techniques 

• Knowledge of the concepts for hiding data (Steganography) 

• Ability to examine and interpret computer forensic artefacts (internet 

history, temporary internet files, email, etc.) to provide a timeline 

analysis of events that occurred on a computer or other digital 

device under examination 

• Ability to analyze advanced or non-traditional compter operating 

systems or proprietary software applications 

• Ability to analyze unique digital devices such as global positioning 

systems or entertainment systems, and 

• Ability to extract data from a live computer or network using 

advanced forensic extraction tools and methodologies Knowledge to 

recognize non-traditional operating systems and proprietary 

software applications, password protected files, and encryption 

• Knowledge of how data is stored on various media types and 

deleted data recovery techniques 

• Knowledge of the concepts for hiding data (Steganography) 

• Ability to examine and interpret computer forensic artefacts (internet 

history, temporary internet files, email, etc.) to provide a timeline 

analysis of events that occurred on a computer or other digital 

device under examination 

• Ability to analyze advanced or non-traditional computer operating 

systems or proprietary software applications 

• Ability to analyze unique digital devices such as global positioning 

systems or entertainment systems 

• Ability to extract data from a live computer or network using 

advanced forensic extraction tools and methodologies (43) 

In 2010, the Scientific Working Groups on Digital Evidence and Information 

Technology (SWGDE/SWGIT) published the Guidelines & 

Recommendations for Training in Digital & Multimedia Evidence to assist 

agencies with designing training programs and to maintain awareness of 

new developments (44). 

3.5 Tool validation 

In 2009 the Scientific Working Group on Digital Evidence published the 

Recommended Guidelines for Validation Testing for all organizations 

406

performing digital forensic examinations (45). The recommendations state 

that validation testing is critical to the outcome of the entire examination 

process. Validation is essential to demonstrate that examination tools, 

techniques and procedures are suitable for their intended purpose. It 

recommends that tools, techniques and procedures should be validated 

prior to initial use in digital forensic processes. Failure to implement a 

validation program can have detrimental effects (46). 

Sommer argues that regulatory trends in forensic science point strongly to 

the need for exhaustive testing of all findings and tools (47). Sommer 

states that the message from forensic science to expert witnesses is clear: 

evidence tendered. should be based on validated scientific knowledge; all 

tools and procedures deployed should have been subject to independent 

testing to demonstrate compliance with their stated claims. He poses the 

question, what happens if the raw material with which you work changes at 

a rate faster than these cycles of publishing and testing? In fields such as 

digital forensics, the rate of technological change is faster than the normal 

times required for peer-reviewed publication. 

4 Digital Forensic Examinations 

4.1 Evidence triage 

Casey submits that missed evidence is a concern with survey/triage 

forensic inspections and preliminary forensic examinations with the 

possibility of overlooking exculpatory evidence48. Forensic examiners 

must respond to all inquiries objectively, and they have a duty to the court 

to report any potentially exculpatory or additional inculpatory evidence they 

find. Forensic examiners are also obliged to reevaluate their findings or 

alter their opinion in light of raised defences. However, there is no ethical 

requirement that the forensic examiner fully investigate any or all potential 

defenses (49). 

Casey submit that if laboratories are to keep up with mounting case loads 

while at the same time keeping current with technology developments, they 

must innovate and adapt to a constantly evolving field (50). 

4.2 Evidence reproducibility 

The Internet is continuously developing with websites being added, 

removed, and continuously updated. Eltgroth states that because of this 

transitive nature of the World Wide Web, there is no certainty that a user 

will be able to access content in an identical form in the future. For this 

reason, the evidential value of data from a prohibited web site found on a 

suspect’s computer may be limited if the prosecution is unable to prove the 

existence of that website and its content at that point in time. 

Eltgroth discusses the application of the Wayback Machine, a service 

provided by the Internet Archive that accesses the largest online digital 

collection of archived Web pages in the world. The Internet Archive 

407

indexes and stores Web pages to allow researchers to access discarded or 

since-altered versions. Such a capability may be an asset for the 

prosecution also to prove the contents of a particular website a specific 

point in time. According to Eltgroth, however, some courts have excluded 

evidence obtained via the Wayback Machine, citing both authentication 

and hearsay concerns. As a result, parties who rely on the content of a 

Web page currently face an unclear evidentiary standard for the admission 

of what may serve as the only available record of since vanished content 

(51). Once a party removes content from a website, the current case law 

fails to provide a clear standard for how to authenticate archived Web 

pages offered in lieu of the missing original (52). The skepticism that courts 

have displayed with respect to archived Web content appears to result 

from a combination of reluctance to attribute archived content to a content 

provider and concerns about accuracy and the potential for intermeddling 

(53). 

Berman states that in order properly to preserve a webpage, it must be 

captured at the exact moment in time in which the preservation is desired 

(54). It may also require multiple captures to produce a series of 

"snapshots," showing how a site changed over time which is especially 

useful in instances where the very fact of habitually updating a website, or 

the dates and times of the updates, are of evidentiary significance (55). 

Once content on a social networking site, blog or wiki has been identified 

as relevant, it must be preserved in such a way that it can later be 

authenticated for use as evidence in litigation, as well as to show that a 

complete preservation was performed (56). Preservation must be done 

completely to ensure that all website information including that which 

might be contradictory to the stance of a party is captured (57). 

4.3 Operating systems 

Sommer states that all operating systems, applications and services are in 

very wide use and within all are forensic traces that have significant 

potential for a very wide range of investigations (58). He states that the 

arguments about speed of change in relation to personal computers can be 

applied with at least equal force to cellphones, where major manufacturers 

release new models with new features and new opportunities for digital 

footprints to be created every 3 or 4 months. He states that digital 

evidence will have to be at least a year perhaps more behind the ways in 

which computers are used by organizations and individuals and criminals. 

4.4 Hand held devices including mobile (cell) phones 

Marshall describes technical assistants such as smart phones as 

“unsleeping witnesses”(59). Stillwagon describes them as portable 

microcomputers embedded in public spaces (60). Even though the device 

itself may not have played any role in the crime, its very presence at the 

crime scene, or use by one of those involved, often leads to it being a 

potential source of information or images of evidentiary value (61). 

408

Navigation devices are increasingly being examined in criminal cases to 

identify data of evidential value. The evidential value of such devices are 

obvious and becoming increasing available with the growth in the 

availability of this technology particularly outside of traditional applications. 

This technology is now a common feature within smart phones to provide 

not only a traditional navigation function but to geographically record the 

location at which a photograph was taken, or report the location of the 

nearest teller machine. Obviously such evidence may provide a strong 

indication between the user and a particular address. In any offence 

where a person’s movements are of interest, the analysis of a GPS device 

to retrieve the stored locations may be evidentiary value. Forensic 

analysis may recover deleted location records and identify where the 

device has been in addition to destinations which may have been entered 

in the device (62). 

4.5 Forensic tools 

Limongelli states that there is a tendency among many proponents of 

open-source software to assert that proprietary software applications 

should be evaluated by courts by different, and higher, standards than 

those applied to scientific or technical evidence generally (63). He states 

that although there is case law regarding the proper standards for the 

admission of technical or scientific evidence, none of that case law 

supports a distinction between proprietary software and open-source tools. 

He submits that if a particular proprietary software is unreliable, the 

appropriate remedy is already embodied in existing case law and the rules 

of evidence. 

Lei pan submits that correctness testing methodology enables the tester to 

achieve testing results of high quality from a manageable number of 

observations and in a dynamic but controllable way (64). This is of 

particular interest to forensic testers who do not have access to 

sophisticated equipment and who can allocate only a small amount of time 

to testing. Lei Pan stated that the focus of the testing should be on the 

verification of the user’s desired features and prioritise frequently used 

features. One concern however with tool validation is the assumption that 

the tester and ultimate end user have operated the tool correctly according 

to the user manual. A further concern for the reliance on tool validation is 

that CFT are using the tool on a wide variety of hardware and software, not 

necessarily the hardware and software employed for the validation 

process. The question may arise “Was the tool validated for the Asus 

Laptop model XYZ? If not, is the tool reliable? A further question also 

relates to the release of new versions of a tool. For example what were 

the changes between version 1.5 and 1.4 and does version 1.5 invalidate 

1.4? The practitioner should expect these questions in cross examination. 

4.6 Managing investigators and prosecutors expectations 

409

Pollitt states that managing expectations is the key to conducting good, 

defensible forensic examinations (65). This he states is achieved by 

learning to clearly articulate digital forensic case objectives and the 

concomitant forensic questions, to gain the understanding and agreement 

of digital forensic clients, and provide realistic feedback on the proposed 

investigative request. He states that digital forensic examiners need to be 

persuasive communicators who can help their clients to more clearly 

understand their own cases and the limits of the examiner’s ability to help 

them. 

5 Legal Issues 

5.1 Presentation of digital evidence 

The case of Daubert obligated trial court judges to assume the role of 

“gatekeepers” and to exclude proffered scientific evidence unless it rested 

on scientifically valid reasoning and methodology (66). The National 

Acedemy of Sciences Report states the least that the courts should insist 

upon from any forensic discipline is certainty that practitioners in the field 

adhere to enforceable standards, ensuring that any and all scientific 

testimony or evidence admitted is not only relevant, but reliable (67). 

According to Neufeld, for years in the forensic science community, the 

dominant argument against regulating experts was that every time a 

forensic scientist steps into a courtroom, his or her work is vigorously peer 

reviewed and scrutinized by opposing counsel. Neufeld states that a 

forensic scientist might occasionally make an error in the crime laboratory, 

but any vigorous cross-examination would expose it at trial (68). 

The National Academy of Sciences Report states that scientific 

conclusions are subject to perpetual revision with the law in contrast 

required resolve disputes finally and quickly (69). Law enforcement officials 

and the members of society they serve need to be assured that forensic 

techniques are reliable. Therefore, we must limit the risk of having the 

reliability of certain forensic science methodologies condoned by the courts 

before the techniques have been properly studied and their accuracy 

verified (70). 

The requirements are not without their detractors. Casey argues that 

although these requirements are designed to raise the bar for forensic 

disciplines, they could have unintended adverse ramifications for digital 

forensic practitioners and labs (71). Casey states requiring digital forensic 

practitioners to be board-certified may be overly restrictive, and may need 

to be broadened to accommodate several digital forensic certifications. 

Casey states that unfairly burdening small local law enforcement and 

private sector labs with accreditation requirements designed for large 

410

government labs could be counterproductive, exhausting their limited 

resources and driving them out of business. 

5.2 Limitations in the Court’s understanding of digital evidence 

Edmund argues that police, investigative and security agencies, 

prosecutors and federal and state courts have not developed principled 

approaches to the use and admissibility of emerging technologies, their 

products and the different forms of expertise and opinion based upon them 

(72). Edmund argued that these developments are unfortunate because 

relatively few of the new techniques have been tested or independently 

reviewed (73). It is inevitable that courts will impose more demanding 

admissibility standards and that those involved in e-forensics, biometrics 

and identification should begin to respond to the anticipated interest in the 

validity and reliability of their systems and opinions (74). 

6 Future trends 

6.1 The future of digital forensics 

Technological advances in virtualisation tools have and will continue to 

create difficulty for digital forensic examiners. Barrett states that we are 

moving towards a dynamic operating environment where organizations are 

streaming applications virtual operating systems from remote servers to 

desktops on an as needed basis (75). The virtual user environment can be 

deleted hence providing a challenge to the digital forensic examiner to 

research and implement methods to find virtualization artifacts and identify 

virtual activities that affect the examination process of certain virtualized 

user environments (76). 

6.2 Investigative management 

SO/IEC JTC 1/SC 27 Information Technology - Security Techniques, 

Guidelines for identification, collection and/or acquisition and preservation 

of digital evidence is an international standard to ensure that responsible 

individuals manage digital evidence in accordance with practical ways that 

are acceptable worldwide, with the objective to preserve its integrity and 

authenticity (77). The International Standard provides guidelines for digital 

evidence management. It describes the processes of identification, 

collection, acquisition and preservation of potential digital evidence that 

may be of evidentiary value. The objective is to assist organizations in their 

disciplinary procedures, and to facilitate the exchange of potential digital 

evidence between jurisdictions. This standard deals with common 

situations encountered throughout the digital management process. The 

potential digital evidence may be sourced from any type of media, and 

refers to data that is already in a digital format. This International Standard 

does not attempt to cover the conversion of analog data into digital format 

(78). 

411

6.3 Evidentiary problems with emerging technologies 

Nelson states that in spite of a concerted effort by law enforcement to 

teach first responders how to properly seize electronic evidence, we still 

see instances where the last access dates of files have been altered by 

untrained investigators or officers looking at the evidence post-seizure 

(79). She states that it appears to be particularly alluring to take a look at 

anything involving sex, but trampling on the evidence in their eager- ness 

to see what they have provides for the defence counsel a fortuitous result 

in which proper forensic procedures were not followed and the dates of last 

access by the defendant are now permanently lost or altered (80). 

6.4 Hand held devices 

The Apple iPhone was released in the US in June 2007 and in late 2009 

Apple released the 3GS version with 32GB storage capacity. The iPhone 

has internet connectivity, GPS navigation, camera and video capability. 

Apple promotes the iPhone feature “Remote Wipe” which can be launched 

as its name suggests remotely by the authorized user to protect their 

private information in the event that the phone becomes lost or stolen, or 

by inference when it is seized by a law enforcement agency. Apple 

promotes this feature stating “Your iPhone contains information you don’t 

want in the hands of a stranger. So if you lose your iPhone and displaying 

a message on it hasn’t resulted in its safe return, you can remotely lock it 

with a passcode or initiate a remote wipe and restore factory settings (81)”. 

6.5 Practitioner welfare 

The University of New Hampshire, Crimes Against Children Research 

Centre published a survey of work exposure to child pornography in 

Internet Crimes Against Children (ICAC) Task Forces and affiliates in 

November 2009. The report concluded that the well‐being of personnel 

who investigate child pornography cases is a matter of concern, not just for 

compassionate reasons, but also because unrelieved stress can lead to a 

reduction of productivity and retention of the specialized and highly trained 

staff who investigate these crimes. It recommended the undertaking of 

further research to systematically examine what characteristics or 

circumstances place some individuals at risk for adverse reactions and 

what agency policies and practices best alleviate negative impacts for 

those doing this important work (82). The report claimed to be the first step 

to answering critical questions about the impact of viewing child 

pornography on law en‐ forcement investigators. The findings indicated 

that some law enforcement personnel suffer ill effects, while others cope 

well (83). Application of this International Standard requires compliance 

with national laws, rules and regulations. The International Standard 

outlines the minimum requirements necessary for enabling transfer of 

digital evidence between jurisdictions. It provides a framework for the 

development of processes and procedures for the identification, collection, 

acquisition and preservation of digital evidence (84). 

412

6.6 Virtualisation 

Davis states that cloud computing has been heralded as a new era in the 

evolution of information and communications technologies (85). 

Businesses using cloud computing services will benefit from reduced 

operating costs as they cut back on IT infrastructure and personnel. 

Individual users will no longer need to pay for software application 

licences. Users will have access to their data from any internet ready 

device in the world. Cloud computing comes with inherent legal risks. The 

global nature of cloud computing raises many questions about privacy, 

security, confidentiality and access to data. For example, Google 

reportedly scans the content of emails to stream “personalised” targeted 

advertising to the end user. Moreover, it is uncertain as to who has access 

to the end user data. It is uncertain how the information can become “deidentified” 

or destroyed. In the face of legal uncertainty, users should rely 

on cloud computing with caution. 

Palmer describes three-dimensional virtual environments (3dves) as being 

the new generation of digital multi-user social networking platforms (86). 

Their immersive character allows users to create a digital humanised 

representation or avatar, enabling a degree of virtual interaction not 

possible through conventional text-based internet technologies (87). In 

addition to the conventional range of cybercrimes, including economic 

fraud, child pornography and copyright violations, the ‘virtual-reality’ 

promoted by 3dves is the source of great speculation and concern over a 

range of specific and emerging forms of crime and harm to user (88). 

These immersive and synchronous elements of 3dves alter conventional 

notions of cyber-harm by converging text, audio and video communication 

methods. Therefore, offensive conduct can encompass interaction through 

speech, gestures, or simulated behaviours including sexual acts, assaults, 

gunshots and even terrorist attacks (89). 

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89 





418

CHEMICAL EVIDENCE 

  419 

Fire cause and fire debris analysis 


Niina Viitala, MSc, Chair of ENFSI/ FEIWG 

Tia Kakko, BSc 

National Bureau of Investigation 

Box 285 

FI-01301 Vantaa 

Finland 

email: niina.viitala@poliisi.fi 

http://www.poliisi.fi/poliisi/krp/home.nsf/pages/indexeng 

420


1.INTRODUCTION  422  

2.FIRE SCENE EXAMINATION  423  

2.1  Site examination techniques and arson dogs  423  

2.2  Sampling and sample packaging  424  

2.3  Vehicle and technical fires  425  

2.4  Fatal fires  426  

3.LABORATORY ANALYSIS  427  

3.1  Sampling and sample preparation techniques  427  

3.2  Methods of analysis  428  

3.3  Individualisation of ignitable liquids  431  

3.4  Statistical methods in analysis  432  

3.5  Spontaneous combustion  433  

3.5  Background matrices studies  435  

4.MODELLING  436  

5.EDUCATIONAL ASPECTS OF FIRE INVESTIGATION  437  

6.EXPLOSIONS  438  

7.CRIMINOLOGICAL ASPECTS OF FIRE SETTING  438  

8.INTERNATIONAL CO‐OPERATION  439  

8.1  Working groups and conferences  439  

8.2  International Collaborative Tests  440  

9.SUMMARY  441  

10.BOOKS AND OTHER PUBLICATIONS  442  

11.REFERENCES  443  

421


Investigating a large fire scene may be demanding as detecting sampling places is 

crucial. Electronic sniffers and arson dogs may be used in detection, but their 

performance has to be evaluated. Validation and the use of electronic equipment 

have been a part of everyday work for a long time. Likewise, there has been a 

discussion about arson dogs and the need for their validation. Vehicle fires have 

received quite a lot of attention recently due to the high number of casualties. 

Reconstructions and modelling are the methods used by fire investigators to 

determine and evaluate the cause of the fire. 

In the sampling field, the main issue has been the use of solid phase micro extraction 

and its developments receiving a lot of academic interest. Two-dimensional gas 

chromatography has gained interest too due to advances made in mathematical data 

analysis. Interpreting chromatograms has become easier with statistical methods. 

Identification and classification of ignitable liquids is one very important aspect of fire 

investigation; chemometrics are also used in this area. The problems of background 

matrice effects have partly been solved by collecting data of fire debris matrices. 

Another analytical aspect in fire debris analysis is the increasing amount of biodiesel 

samples. The challenge lies in two different components of biodiesel, namely 

vegetable oil and hydrocarbon, which require different methods for analysis. 

Spontaneous combustion also needs to be taken into consideration while handling 

biodiesel samples. 

Understanding firesetters’ behaviour is important in order to facilitate solving arson 

crimes. Most of them are young; studies indicate that the younger the person is when 

setting their first fire, the more likely it is that he will become a repeat offender. 

Alcohol abuse, emotional and behavioural problems in childhood as well as 

intellectual disabilities are often found to have a connection with multiple fire settings. 

A danger of terrorism cannot be underestimated either, as pyro-terrorism has 

become a tactic of choice for terrorists. 

International co-operation is a very important way of sharing resources in forensic 

science. Many countries have only one forensic laboratory and only a few members 

working in the fire investigation group limiting their possibilities for development work 

in all fields of fire investigation. Different kinds of proficiency tests are a part of the 

collaboration for forensic scientists with limited resources to develop their work. 

There is a large review article about fire investigation spanning the years 2001-2007 

covering various subject matters, references to which are made in many sections of 

this paper. 1 

422

2. Fire scene examination 

2.1 Site examination techniques and arson dogs 

A fairly modest amount of work has been done for improvements in fire field 

investigation during the last three years. A practical guide emphasizing the 

significance of a good and thorough fire scene investigation has been published for 

first responders. A proposal for doing site examination in a specific order, including 

the correct documentation consisting of photography and sketches with dates and all 

important information, is essential for securing quality of the work. Being aware of 

one’s own behaviour at a fire scene is crucial in order to avoid damaging or altering 

the scene. Preserving evidence is very important for final fire investigation in order to 

determine the origin and spreading of the fire throughout the scene. 2, 3 A thorough 

compilation of fire patterns includes an overview of both historical and recent 

research. It emphasizes the need for burn pattern analysis in respect to pattern 

reproducibility and formation airflow including ventilation studies. 4 

The selection of fire debris samples can be challenging due to destructive processes 

occuring during a fire. A study about a liquid fuel on carpet was conducted in order to 

aid the selection process. The aim was to reveal the physical processes of and 

indicators left by such a fire. The article largely questions the current practices 

involved in fire site examination and proposes scientists to undertake research work 

to advance the role of science in fire investigation. 5 A book about fire debris analyses 

includes a chapter focusing on detection of ignitable liquid residues at fire scenes 

with colour pictures of burn patterns where ignitable liquids have been used and 

comparison pictures where they have not been used. Principles of electronic sniffers 

and colorimetric tubes are also demonstrated including both the benefits and 

disadvantages of the methods in question. The operating principle of ignitable liquid 

absorbent and ultraviolet detection as well as the use of these methods is also 

considered. 6 

Portable field devices — misleadingly also referred to as electronic noses — have 

been studied in a few articles. Fluorescence-based vapour sensors have been 

designed and tested. The results of analyzing petroleum distillates in different 

experiments seem promising, although more studies are needed to prove the 

usefulness of the devices. 7 An artificial nose using optical fibre arrays to sense 

vapours has also been developed. Fibres are functionalised with analyte sensitive 

materials, which are also cross-reactive. That enables the use of a pattern 

recognition algorithm and thus the identification of a large amount of different 

vapours including ignitable liquids. 8 Multiple electronic noses were compared in a 

review article and the article lists even the technologies they use and their 

manufacturers. The article discusses the methods, their effectiveness and 

applications from food factories to explosive detection. Future trends are also 

discussed. 9 A Vapor-Sensitive Microsphere Array application that has been used in 

the detection of volatile organic compounds and nerve agents was explored in a 

study to detect ignitable liquids. Despite obtaining high accuracy in distinguishing 

three different ignitable liquid classes in burned and unburned samples, additional 

studies are needed in order to challenge the system with more realistic, simulated fire 

debris samples. 10 

423

A few articles related to arson dogs have been published during the recent years. As 

accelerant detection dogs have become more widely used, validation of the training 

and the dogs’ ability to work have become increasingly important. 1, 11, 12 A standard 

assessment technique is useful for comparing the effectiveness of different dogs, 

breeds, training methods and agencies. It has been demonstrated that assessment 

of behaviour is noth objectively and subjectively important 13 A study was also 

conducted to compare arson accelerant dogs and solid absorbents. According to the 

study, sensitivity of ignitable liquid absorbent was not effective enough to reveal 

either paint thinner, camp fuel or gasoline from floor panels (the term gasoline is 

used in this report instead of the British term petrol), whereas the detection dog 

alerted 16 out of 18 panels. 14 In another case, the evidential value of ethanol was 

pointed out with an arson dog; the canine unit was used to demonstrate sampling 

positions. The results showed ethanol in samples and thereby the complainant’s 

explanation for the fire was implausible; the indidual confessed to setting up the fire 

with the commercial product ethanol. 15 

Hypotheses may be formulated in borderline cases where conclusions between 

chromatograms and dog’s behaviour is not self-explanatory. In these cases, 

collaboration between scientific analysts and canine accelerant detection teams is 

extremely valuable. In all cases where accelerant is detected in a sample by a dog or 

via another method, the sample must be analysed in a laboratory for formal 

identification and confirmation of the results. 16 An article concerning standardization 

of the common methods in forensic science also addressed the sources of standards 

and how the standards compared to ones in quality assurance schemes. 17 

A two-part series touches upon intelligence-led crime scene processing. The first part 

gives instructions on how to allow forensic science to become involved in security 

assessments by means of forensic intelligence. 18 The second part integrates crime 

scene investigation into the intelligence thinking model. New perspectives for 

investigations are introduced when the examination process is conducted in a 

different way. 19 

2.2 Sampling and sample packaging 

Although sampling is one of the most important issues to consider in fire investigation, 

literature focusing on sampling is scarce.. Mistakes made in sampling cannot be 

corrected in further fields of sample analysis. Samples from a fire scene must be kept 

in a vapour tight and non-contaminant container. Different kinds of commercial 

containers have been studied to reveal their hydrocarbon leaking rates. 20 

Some studies have been made on the topic of gasoline on hands. Background 

effects and efficiency in the collection of gasoline have been evaluated by using 

different kinds of gloves. Statistical methods are included in the analysis in order to 

evaluate variations in the results. According to the results, the suspect should wear 

gloves for approximately twenty minutes after which the gloves should be placed in a 

vapour tight packaging. A prototype sampling kit was developed for the purpose with 

optimal support. In order to minimize contamination, the sampling kit should have 

424

instructions on the label on the top of the kit. The developed kit has been considered 

21, 22 

to be practical and easy to use. 

2.3 Vehicle and technical fires 

Car fires may be divided into different types according to the origin of the fire. Fire 

debris can be used as a comparison sample or as trace evidence. Ignitable liquids 

found close to a burned car can be used as evidence to prove that igniting the fire/car 

was intentional. This report covers different articles on the topic. Firstly there are 

cases with casualties, either suicidal or accidental 23, 24 , secondly malfunctions of 

electronic or technical apparatus 3, 25 and thirdly evidence gathered from a burned 

vehicle. 26 

Security improvements in cars during the last decades have simultaneously altered 

both the amount and the quality of combustible materials. For example, plastics 

constitute the major fire load instead of gasoline, being the primary cause of death in 

car crashes. The fire progresses rapidly depending on the direction of the impact and 

the amount of combustible plastics. Foam surfaces and fabrics change the direction 

in which the fire is spreading while the passengers are stagnant, which is why 

survival time inside a flammable vehicle is usually less than 10-15 minutes. 27 

Formation of different kinds of gases plays a significant role in car fires. For instance, 

inhalation of chlorine or carbon monoxide gases has been reported to be the cause 

of death. The origin of such gases is either the covering of a passenger compartment 

or a supplementary heater used in a cabin of a truck. Malfunctions in heaters 

generate carbon monoxide while the heat transfer to the vicinity of a heater might 

23, 24, 28 

also cause a fire, subsequently leading to poisonous fumes being released. 

Vehicle reconstructions have been used to help fire investigators determine the 

cause and origin of car fires. Evolutions of fire were controlled by thermocouples and 

the fires were photographed and recorded. Furthermore, fire patterns were used to 

analyse the origin of the fire. Previous assumptions about vehicle fires and their 

developments, evolutions and venting were dispelled. 29 

Functional problems of electronic and technical apparatus in vehicles are often 

difficult to detect due to severe damage to the structures. An individual case case 

might not be serious, but occurrence of more than one similar incident can be 

dangerous. An example of such cases would be automobile recalls. One cause has 

been e.g. leakage of gasoline from a fuel line, as polymetric materials might build up 

an electrostatic current and therefore create a small hole in the fuel line. Daylight 

running modules have been in poor contact between pins and sockets, producing 

overheating and thus fire even when the car has not been running. 3 Another, 

different case study focuses on metallurgical analysis to detect the origin of the fire 

on a bus. The fire occurred while the bus was travelling at a highway speed. 

Inspectors had two different theories for the cause of the fire; either a dragging brake 

or an overheated underinflated tyre. The method of analysis consisted of 

microstructural analysis and hardness tests. One tyre from the bus was used as a 

comparison reference in this case. The metallurgical analysis showed that the 

dragging brake was the origin of the fire. 25 

425

A burned vehicle and its interiors may be used as fingerprinting evidence in forensic 

work. The geological matrices inside the car can be studied and compared. A study 

has been conducted on examining quartz grains after they have been exposed to fire. 

The article demonstrates that while soil and sediment alter in car fire temperatures, 

most quartz grains are not altered in temperatures below 900°C. As a result, 

thoroughly studied grains can be used in forensic work. 26 

As mentioned before, a failure of a mechanical or electronic component may cause 

fire. Ignitions based on mechanical origins are varied, including friction and further 

sparks, overheating, abrasion of electronic wires, leakage of flammable gases and 

corrosion. Electronic sources of ignition are also multiple, consisting of arcing, 

resistance heating (e.g. by poor connections), external heating due to a rupture in 

3, 30 

insulation and a discharge of static electricity. 

A suggestion for the technical improvement of the detection area has been 

demonstrated in a meeting. Scanning Electron Microscope (SEM) has usually been 

used for locating metallic damage with a conductor and excluding other matter. A 

comparison between SEM and an alternative method, a confocal laser scanning 

microscope, has been conducted by examining dozens of electronic conductors 

gathered from fire scenes. 31 

2.4 Fatal fires 

More than a million people die each year as victims of suicide or homicide. Some of 

these cases are straightforward and obvious, but some challenge investigators in 

detecting the cause of crime. Depending on the country, its region and culture, the 

method of fatal death varies greatly. For example, burning is the most common 

suicidal method in females in India. 32 In the Adana region of Turkey, nearly seven 

per cent of deaths due to burns are criminal in nature. Homicide cases are committed 

either by torching of homes or arson with kerosene whereas fire accelerants are used 

in cases of suicide. 33 In Germany, on the other hand, the number of self-cremation is 

on the rise. Using an ignitable substance is also common there. The number of 

psychotic patients is high and victims have often attempted suicide earlier on. 34 

There is a report on catastrophic fires that claimed multiple victims in the United 

States in 2008 covering structural and non-structural fires and determining roles of 

smoke alarms and sprinklers. 35 

One extreme unsettled fire cause is ignition of a human body where the body often 

has significant burns on the abdomen, upper legs and chest. No apparent source of 

ignition exists and generation of the fire seems to have been spontaneous. 

Furthermore, the surroundings of the body are often unharmed. Despite various case 

reports about this matter, the general mechanism of the fire seems to be a modest 

external flame source which inflames clothes or other suitable substances and 

subsequently releases subcutaneous fat. 36-38 

Fatal fire scene investigation has been developing towards archaeological 

techniques due to the possibilities they offer of maximising evidence recovery and 

interpretation. 39 Archaeological data collection methods are systematic, consisting of 

grids or zones and starting from the least destructed areas in order to avoid scene 

disturbance. The use of alternative light sources can be used in searching the scene. 

426

39 The distinction between natural and unnatural death is based on the careful 

collection and correct interpretation of forensic findings. 34 It is important to 

understand the fire incident and its by-products, and how they affect the human body. 

It is recommended that the body is examined, with all due respect to the deceased, 

at the fire scene in order not to lose any crucial evidence. Documenting the fire 

debris found above and underneath the body is also necessary due to determination 

of incapacitation of the victim. 40-42 

A different area is toxic gases released during the fire and further during the heat 

below the combustion temperatures of 200 °C. Carbon monoxide is usually assumed 

to be the cause of asphyxia and thus other gases are neglected. Nitrogen, carbon 

monoxide (CO) and hydrogen cyanide (HCN) from polyurethane are identified by the 

cryogenic mode of GC-MS. A novel sampling system is also designed. 43 A 

reconstruction of a hospital fire is also made to demonstrate the requirements for 

materials used in general facilities. Flooring material consisting of polyurethane 

based mattresses, which does not ignite easily, but after ignition it burned intensely 

producing hydrochloride, isocyanate and carbon monoxide etc. All of the 

aforementioned gases might have been the cause of death, but only CO and HCN 

were actively looked for as they belong to the normal autopsy procedure. 44 

3. Laboratory analysis 

3.1 Sampling and sample preparation techniques 

During the last decade development of solid phase micro extraction fibres (SPME) 

has been under discussion. Fused silica has been the key component of the 

substrate to which the coating has been applied. Unfortunately, the base of the silica 

is very fragile and thus its use is limited. 45 Studies concerning SPME methods focus 

on the issues related to either substrate or coating development. 

Recent progress in SPME fibres has been the introduction of elastic NiTi assemblies. 

This rigid titanium structure has been combined with polydimethylsiloxane (PDMS), 

sol-gel and silica particles to increase mechanical durability and extraction capacity. 

A study was conducted on the performance of extraction, repeatability, reproducibility 

and extraction kinetics. Even potential applicability was explored with a fire debris 

analysis. The study found that, when compared with commercial fibre, the methods 

gathered identical chromatograms. The absolute peak magnitudes carry the 

differences in the methods. 45 In addition, there have been a few studies focusing on 

a home-made SPME fibre. The fibre was coated with 1:1 molar ratio of 

octyltriethoxysilane (C8-TEOS). Methyltrimethoxysilane (MTMOS) and a 

polydimethylsiloxane divinylbenzene (PDMS/DVB) fibre was used as a comparison. 

Selectivity and extraction capacity of the fibres were determined. Selectivity of C8fibre 

was good and enabling the extraction of hydrocarbon compounds from 

simulated arson samples with better extraction efficiency than the reference fibre. 46, 

47 

In another study, a direct contact is made between solid phase micro extraction fibre 

(SPME) and the fire debris. 48, 49 This was made possible by an improvement for 

recovery of low volatility components in the sample. The method was discovered to 

427

e suitable for use in screening in forensic work. 49 Another study reported that 

SPME is an effective and solventless technique for recovery of accelerants from fire 

debris. 50 

Singed hairs in arson cases provide a different kind of evidentiary value. When using 

volatile accelerant for setting up a fire, hair on the back of an arsonist’s hands and 

head is damaged due to deflagration. A sampling method for such cases has been 

studied. In the study, samples were taken with a special razor previously proven to 

be effective in routine investigations. Stereo and transmitted light microscopy was 

used in analysing the samples to search for evidence of heat damage. The results 

show that the damage caused to the hairs could last up to two months. As only a few 

hairs in an area might be singed and others can be undamaged, they should be 

collected in a thorough manner with particular attention paid to the fingers. 51 

3.2 Methods of analysis 

Analytical methods are divided into three sections: comparison of samples, 

comparisons of analysis methods and (novel) techniques. In some cases it is difficult 

to define the appropriate section for the subject of a study. 

In order to obtain qualitative results in a fire debris analysis and the related extraction 

process, the use of an internal standard is suggested. Characteristics of the standard 

should involve a few key features. Firstly, the compound needs to be a compound 

not commonly found in fire debris samples; secondly, it needs to be eluted within the 

same time frame as the target compounds; thirdly, the mass spectrum of the 

standard should be different enough to be distinguished from the sample; and lastly, 

the cost of the internal standard should be reasonable. 52 The study further 

recommends that the internal standards be delivered into the fire debris in the same 

solvent that is used to elute the charcoal strips or that the standard is placed in a 

paper that is subsequently added into the fire debris can. In the matter of evaluating 

the standards, the study suggests that a combination of two different standards be 

used in order to obtain the whole range of boiling points, i.e. compounds from both 

low and high boiling point areas. 52 

Comparison samples are often present in fire investigation due to their revealing 

aspect. Sometimes the aim is to demonstrate the use of a certain commercial 

chemical or substance. 53 Alternatively, the aim might be to establish the evidence 

value of a compound. In such a case, thorough knowledge of the sample and its 

composition is crucial. 54 A number of publications touch upon this topic. In one often, 

jet fuels are classified by using fast gas chromatography coupled with a fast scanning 

quadrupole ion trap mass spectrometer (GC-QIT MS). The method was applied in a 

two-way data collection mode. After transforming the data with wavelet and principal 

component transformation, the fuzzy rule building expert system (FuRES) and partial 

least squares (PLS) are used for classification. These mathematical methods were 

employed due to their reproducibility and adaptability to interpretation. 55 

A large survey was conducted on Canadian gasolines. Seventy-six (76) samples 

consisting of regular, mid-range and premium unleaded gasolines were collected in 

2004 from 12 different retailers around Canada. GC-MS was used as a method of 

analysis and the data was processed with Agilent Chemstation software. In the article 

428

considering examining the total ion chromatograms (TICs), extracted ion profiles 

(EIPs) and ASTM methods have generated discussion. The so-called average 

gasoline pattern which can be applied to case work was a point of interest. The 

examiner, on the other hand, might notice an exceptional pattern that can produce 

difficulties in identification the substance as gasoline originated. A variety of refineries 

contributes to the diversity of chemical compositions in gasolines. The article 

concludes that two ASTM methods (1387 and E1618) can provide general guidelines 

for forensic work. 56 

In relation to the above-mentioned subject, discrimination of diesel oil by origin and 

type is also discussed. Methods are physicochemical instead of gas chromatographic, 

and results are valuated by mathematical methods, such as principal component 

analysis and linear discriminant analysis. 57 According to the study, determining the 

origin and type of a diesel sample is possible without the use of complimentary 

assays. On the other hand, grouping by composition is not distinguished. 57 

Furthermore, spectrofluorimetric methods are used in order to determine the residual 

oil in diesel oils. There the mixture of fuels is under research due to the obligatory 

commercialization of biodiesels. The synchronous fluorescence spectra are able to 

categorize diesel oil samples and make groupings by using linear discriminant 

analysis. 58 In the same series of studies, gasoline adulteration is identified by using 

comprehensive two-dimensional gas chromatography. A generic method is 

developed in order to assess the adulteration of gasolines; this method of analysis 

can be a powerful tool in solving similar analytical problems with complex mixtures. 59 

Since the use of base stock in motor oils has many applications, the possible 

association of a motor vehicle in arson is probable. It is in these cases that 

comparing the oils becomes relevant. Some investigations have been made into 

comparisons of fuel oils and motor oils. Distinguishing several different oil brands and 

grades from each other is possible by using high temperature gas chromatography 

mass spectrometry (HTGC-MS) with a programmable temperature vaporizer injector. 

Increasing viscosity lengthened retention times and while “the chromatogram 

envelopes” broadened identification of the samples whereas unused oils from the 

same brand and viscosity could not be distinguished. 60 In another example, 

composition and a weathering pattern of heavy fuels is analysed based on IR, GC- 

FID and GC-MS analyses 54 , and further with high temperature two dimensional gas 

chromatography HT-GC x GC 61 and easy ambient sonic-spray ionisation mass 

spectrometry. 62 

In a Japanese study, detection of total sulphur in fuels demonstrated that different 

fuel types could be distinguished by using gas chromatography with atomic emission 

detection (GC-AED). Due to the high degree of selectivity and sensitivity in GC-AED 

and detection of characteristic elements in compounds, detection of trace amounts of 

substances in fire debris samples is possible. Furthermore, although the distillation 

characteristics had altered, discrimination of fuel types was successful. 48 

A novel concept of isomer abundance analysis (IAA), supersonic gas 

chromatography mass spectrometry, has been used in order to widen the range of 

fuel and hydrocarbon analysis. The method reveals molecular ions in all 

hydrocarbons and this helps to obtain their isomer distribution and provide 

identification for them. Due to the trace amounts of fuels collected from fire scenes, 

429

sensitivity of the method is important. In diesel fuel characterization the Supersonic 

GC-MS achieved the ppb -level (ng/ml). Biodiesel samples were also studied but the 

analysis was made without pre-treating the samples. The molecular ions of the 

samples were discovered. The method was found adaptable to use in forensic 

work. 63 Another type of separation capacity is considered when GC-MS is used in 

analysing alkylcyclohexanes. The purpose of the method was to determine whether 

the residues found in the fire debris were kerosene, gas oil or plastic based. 64 

Another suggested method for the characterization of gasoline samples is multidimensional 

GC-fourier transform ion cyclotron resonance MS analysis (GC-FT-ICR- 

MS). The author of the study refers to fingerprinting crude oils and refined petroleum 

products. Therefore the use of chemical ionization, with protonated reagent ions, can 

be used to identify the presence of co-eluting isomers in gasoline samples. When 

necessary, the method includes dimensions that can be added to the analysis in 

order to achieve greater accuracy. 65 

A different kind of detector for forensic work, ion mobility spectrometry (IMS), has 

been evaluated and developed. An article and a review written about the method, its 

history, ion source development and applications serve to deepen the know-how of a 

fire analyst. The method is expected to play a significant role in ignitable liquid and 

66, 67 

arson investigations. 

In addition to the study mentioned in the previous section, there have been two novel 

studies made; firstly, an application of gas chromatography differential mobility 

spectrometry (GC-DMS) with two-way classification of ignitable liquids in fire debris. 

The GC-DMS provides unique profiles for different ignitable liquids and, combined 

with the fuzzy rule building expert system (FuRES) on two-way data objects, the 

system was successful in forensic analysis of fire debris. Also the method seemed 

promising for onsite detection for arson investigation. 68 Secondly, the same writer 

has published a comparison of different detection methods in order to value their 

performances in ignitable liquid classification by using gas chromatography coupled 

with either differential mobility spectrometry or mass spectrometry, GC-DMS and GC- 

MS respectively. The comparison of the methods was made with projected difference 

resolution (PDR) and the results were furthermore verified by FuRES. One-way 

datasets were determined in order to calculate minimum resolution values. The 

results were that MS data achieved higher resolution, but DMS gained more 

informative data. The two way (2D-GC) datasets in neat samples demonstrated that 

GC-MS would have had smaller prediction errors than DMS in classifying the 

ignitable liquids. No statistic difference occurred between the two methods 

considering the high resolution values of neat and spiked samples, yet with burned 

samples a smaller prediction error was obtained with GC-MS. 69 

Detection of gasoline and paint thinner directly from common household material has 

been studied. The analysis method is a direct analysis of real time mass 

spectrometry (DART). The method does not need sample pre-treatment. Samples 

are introduced at atmospheric pressure and placed into a stream of exited gas, from 

which analytes are desorbed and ionized. The method has been demonstrated to be 

sensitive and chemical fingerprinting was obtained when samples were analysed 

within 1-24 hours after being spiked. Further studies considering matrices and related 

effects for detection are proposed. 70 DART is also mentioned in a review article 

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about forensic applications of ambient ionisation methods. The method is capable of 

detecting compounds in a wide range with minimal sample workup, yet the time of 

analysis is short. Also here the development of the technique(s) is crucial for it to 

become widely used. 71 

Another detection method using mass spectrometry is a proton transfer reaction, 

PTR-MS. The technique has been developed almost exclusively for detection of 

gaseous organic compounds in air, but the possibility of detecting fire accelerants 

onsite has been shown. It is also possible to identify substances after a prolonged 

storage of burnt material with a characteristic mass spectra coupled with principal 

component analysis. 72 

The high content of ethanol in motor fuels may produce problems in fingerprinting 

samples. Especially aliphatic alcohols with the carbon number C3-C8 provide 

challenges for the analytical method used. A flow switching GC x GC has been 

demonstrated to generate high speed and high resolution separations as well as to 

gain quantitative and accurate results. The fast runtime combined with separation 

efficiency seems to surpass the compared ASTM methods. A drawback of the 

method is that it is not capable to separate ethers from gasoline aliphatics, although 

they are not normally added to the gasoline under study (E85). 73 The presence of 

foreign material improperly mixed to samples can be detected by fingerprinting 

applied to chromatographic data. In addition to multi-way calibration techniques 

combined with GC x GC-FID can reveal complex mixtures being treated as a single 

analyte. 59 

Outside the scope of general analysis methods, a study about gasolines and their 

modifications during evaporation and burning by normal phase HPLC has been made. 

The task was to determine polycyclic aromatic hydrocarbons, because they are less 

subjected to alterations during the fire. Arenes were identified not only by retention 

time but also by distribution of absorption spectral lines at peak points. Relative 

concentration of alkylarenes was experimentally obtained. 74 

Another method not so common in arson analysis is stable isotope profiling of burned 

matches. Burned matches may be recovered from fire scenes, but unfortunately the 

isotope profile of matches with gasoline from mock fires were not significantly 

different from the ones without fire accelerant. 75 

3.3 Individualisation of ignitable liquids 

Identification and classification of fire accelerants in arson residues, especially 

ignitable liquids, has gained more efforts each year. The method used to obtain 

distribution varies, although in many cases analysis is made by GC-MS combined 

with different types of data handling. For example, comprehensive two-dimensional 

gas chromatography combined with mass spectrometry has been a preferred 

technique for analysis of complex volatile and semi-volatile mixtures. The use of two 

different types of columns and a modulator offers an orthogonal separation system 

and also provides the target compound’s carbon number and chemical structure. 76 

Alteration of fire debris samples due to microbial degradation of sample matrices 

might lead to differences in chromatographic profiles of ignitable liquids. 

431

Biodegradation occurs rapidly in soil, rotting wood and other organic matter. Because 

fire debris samples are often stored at room temperature for a prolonged time, the 

study concentrates on exploring biodegradation as a function of time. Findings 

include a loss of n-alkanes while branched remained unchanged. Key components of 

gasoline were also degraded; among monosubstituted benzenes, propylbenzene 

was the most vulnerable. 77 Another study about biodegradation considers biodiesel 

mixtures in the sea. In that initial study, fatty acid methyl esters were degraded at the 

same rate as n-alkanes, and more rapidly than other hydrocarbon components, being 

indistinguishable within weeks. 78 

Multivariate pattern recognition is one aspect that has attained attention in sample 

identification. The nature of petroleum-based accelerants is complex and this already 

generates problems for analysis. Yet weathering, microbiological degradation and 

continually changing product formulations of ignitable liquids complicate identification 

even more. Thus an application that uses GC-MS technique combined with 

multivariate pattern recognition techniques is examined to match the patterns in 

weathered and unevaporated fuels. 79 

Another, important study focuses on two relevant matters, namely optimization of 

sampling by SPME and classifying ignitable liquids. The comparison of the 

suggested classification system with a prior one has also been estimated. Data 

obtained from GC was pre-processed and normalised and further processed with 

mathematical models: principal component analysis (PCA) and soft independent 

modelling of class analogy (SIMCA). The data of the study showed that results from 

SIMCA may be used as a predictor of accelerant class and that the samples did not 

show any correlations to other classes. 50 As an alternative process for categorisation 

of commercially available ignitable liquids, classification is made by summed-ion 

mass-spectrum. A spectrum is created by summing the intensity of each ion across 

the entire chromatographic range and furthermore normalizing the resulting spectrum. 

The benefit of the method is that it removes laboratory-depending chromatographic 

profiles and provides a set of characteristic m/z ions for sample identification. The 

data set can lack some portions of the components only if the samples were 

analyzed with very different conditions. The suggested method is the ASTM E 1618 

80 protocol where probability to obtain a correct match is high. 81 

3.4 Statistical methods in analysis 

During the last three years, statistical or chemometric analysis methods have 

become more important in data interpretation. This includes association and 

discrimination of ignitable liquids with the principal aim to eliminate errors in a visual 

comparison of the chromatograms. These methods may be used on group samples 

based on chemical similarities by statistically quantifying the matching pattern. The 

studies related to this period of time have been made by gas-chromatography mass 

82, 83 

spectrometry (GC-MS) as it is a routine method when analyzing ignitable liquids. 

Three different studies have been made on diesel samples because diesel is 

chemically more complex than gasoline. All of the surveys applied data pretreatments, 

such as retention time alignment, normalization and mean centring. 

Samples were analysed in triplicate to ensure reproducibility of the analysis. The 

Pearson product moment correlation (PPMC) was used to analyse correlation 

432

etween pairs of diesels whereas principal component analysis (PCA) was applied to 

the complete data set to assess variation among the diesel samples. 82-84 The results 

of the studies were that one got a strong correlation of PPMC, but had some 

retention time troubles, reminding that care must be taken in data interpretation, 83 

while the other study succeeded to use PCA in identification natural clusters of 

similar diesels and the most discriminatory chemical compounds in the data set 

stating that PPMC could be applied in forensic arson investigations. 82 The third 

examination included temperature programming of the gas chromatograph in the 

PPMC and PCA studies gaining adequate results for use in forensic work. 84 

A different approach to statistical methods is covariance mapping by t-tests, in which 

power analysis is performed to estimate the number of analyses needed to 

discriminate between two samples. As comparative forensic analysis may link an 

individual person to a crime in this method, it is crucial to keep the power of the test 

high. This means that using a significance level of 5 % the risk of type I error is 5 %. 

The error type II occurs when a null hypothesis was accepted incorrectly. In practise, 

the significance level of the tests is dependent on the consequences and keeping 

them low. 85 

Liquid gasoline samples from fire scenes can be subjected to various conditions. 

That brings along variation into samples when comparing them to ones purchased 

from service stations. Multivariate analysis for fire debris casework has been 

developed. Dimensions of gathered GC-MS data was reduced depending on the 

method into two or three, four and ten in canonical variate analysis (CVA), orthogonal 

canonical variate analysis (OCVA) and PCA, respectively. In this study the 

procedures were tested with advance knowledge of the correct assignments. The 

method needs to be studied further in order to have an applicable method to fire 

debris casework. 86 

Sometimes comparison of two different methods and their ability to reveal information 

needs to be evaluated. A method for comparing two methods has been published. In 

the study the within and between -object variances and covariances were estimated. 

Reliability of the two methods was estimated in addition and the parameters were 

evaluated with a performance likelihood ratio model. A similar kind of comparison can 

be made between different kinds of databases, although care must be taken while 

accepting data to the method. 87 

3.5 Spontaneous combustion 

This subject consists of various substances starting from coal industry, saw dust, 

cotton and laundry fires. A major part of recent studies relate to vegetable oils and 

biodiesel and their autoignition. Fundamentals of auto-oxidation relate closely to 

thermodynamics and the surroundings of the self-heating material. 1, 88 In oil tank 

fires oxidation of iron sulphides may lead to tank fires and even explosions. 89 In 

another case, the combustible material was Auricularia auricular, dried mushroom. In 

2003, a warehouse filled with agaric, cinnamon and aniseed was the starting point of 

a severe, fatal fire and the reason is still undetermined. Risks of this kind of materiasl 

are underestimated. 90 

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Although alternative fuels have been a marginal part of fire debris analysis, in the 

near future the amount of samples is supposed to increase. An introduction to 

different types of biodiesel fuels, their sample preparation and analysis has been 

published. 1, 91 In a study of spontaneous ignition of biodiesel samples, it was found 

that the autoignition temperature of biodiesel is much lower, being in isothermal 

conditions around 100 °C, than in vegetable oil. The inner temperature of the sample 

rose rapidly with generation of smoke and the flash point of biodiesel is dependent on 

92, 93 

the concentration of methanol originated from the manufacturing technique. 

The following studies about the deposition of vegetable oils and biodiesel samples 

need to be noted. When dealing with fire debris samples, extraction of ignitable liquid 

residues needs to be carried out first. Fatty acids of vegetable oils are not disturbed if 

samples are heated in 65 °C for 16 hours. Solvent extraction is needed to recover 

vegetable oils, but fatty acid methyl esters (FAME) are volatile and can be analyzed 

by a passive head space. Heavy gasoline distillate chromatogram changes when 

FAMEs are present; in such a case the identification of every extraneous peak is 

essential in order to detect the presence of biodiesel. Another challenge with 

suspected vegetable oil in fire debris are storage conditions of fire debris samples, as 

they should be waterless and the samples sealed well to reduce degradation. The 

91, 94 

ageing of such samples, including biodiesel samples, must be studied further. 

An analysis method for vegetable oil from fire debris has been developed. Different 

derivatization methods were optimized in order to convert fatty acids into esters. 

Furthermore three various columns and different GC programs were used. Base 

catalysed trans-esterification derivated fatty acids in vegetable oils successfully and a 

polar column separated the peaks clearly. The study serves as a starting point for 

research in fire debris in respect to vegetable oils. 95 A fast sampling and analysis 

method of another kind is solid-phase microextraction (SPME) used with a gas 

chromatograph. In this method, thermal desorption of unsaponifiable substances was 

carried out first. Then the remaining oil in the fibre was recovered with solvent, further 

base-catalysed trans-esterification and injected to GC-MS. The analysis was made 

by using two different gas chromatographs with proper columns in the oven. This 

method provided more useful information than the reference method. 96 

Self heating tendency and thermal stability of vegetable oils have also been studied 

with differential scanning calorimetry (DSC). Being a fast and useful technique, it is 

possible to use this method when a sample of questioned oil is available from the fire 

88, 92 

scene. 

An analytical method of C80 calorimeter was used to identify reasons for 

spontaneous combustion of cotton. Calculating the thermal behaviour of cotton and 

its self-accelerating decomposition temperature, variation of results with different 

parameters demonstrated that water plays a key role in the process. Further 

consideration seems to be that microbiological degradation of cotton generates 

oxygen and methane which could lead to ignition. 97 

Spontaneous combustion of laundry has been unrecognised as a fire hazard. Ignition 

of laundry from restaurants or hotels with stains some customary cooking oils has 

been investigated as to tendency to ignite cotton material. The tests showed that the 

prevalent temperature level together with the oil’s properties influences the auto- 

434

ignition. Therefore aims have been modified to develop a proper method for studying 

chemical spontaneous ignition. 98 In another study, this has already been made by 

using a Semenov Model to evaluate the possibility of spontaneous combustion in 

tumble dryers. This model uses steady-state treatment and considers the body of the 

reactor uniform, thus being the simplest thermal ignition model. Two kinds of 

calculations were made in the study: geometry and heat losses for typical dryers and 

flow through the tumbling fabric in the drum. The obtained critical temperature 

predictions showed that the temperature required for ignition would be approximately 

twice the normal operating temperature. This led to the conclusion that in order to 

ignite, external heating for several hours or contamination with a reactive material 

that would self-heat at lower temperatures would be needed. 99 

Spontaneous combustion of Refuse-derived fuels (RDF) has been studied widely. 

Just two studies are mentioned here. Some accidents caused by spontaneous 

ignition of RDF, even with fatalities, have served as initiatives for studying the 

behaviour of refuse-derived fuel in storage conditions. A chemiluminescence 

technique has been used to investigate the phenomenon in low temperatures. The 

stated reason for chemiluminescence is cellulose in RDF. During the preparation, 

RDF was treated with high and low temperatures, and thus cellulose undergoes autooxidation 

producing hydroperoxides, which accumulate in RDF, making spontaneous 

combustion possible. 100 In another study, various conditions including moisture 

content, initial temperature, particle size and the presence of micro-organisms was 

examined. The conditions contributed to the ignition in the sense that a rise in water 

content reduced the induction time, the particle size of RDF sample showed that 

ignition time was shorter in samples that had not been grounded. No relationship 

between bacterial fermentation and spontaneous ignition was observed. 101 

3.5 Background matrices studies 

In forensic fire investigation it is important to compare the analysis results with real 

commercial products. This could be the case with detergents which cannot be used 

as fire accelerants due to their water content, in spite of including the compounds of 

flammable liquids. 53 To prove the likelihood of presence of gasoline residues in the 

shoes or clothes of an arson suspect can be necessary. In a pilot investigation, it was 

demonstrated that although a person has had regular activities with gasoline during 

their work shift, getting a detectable amount of gasoline from the samples is unlikely. 

If the action has been lawn mowning, on the other hand, or a noticeable amount of 

gasoline has been spilt on the clothes or shoes, it is possible to obtain traces of 

gasoline or its components. 102 

Another side in a view of forensic characterization is having the same kind of product 

that is just manufactured in a different way. For example, this might unfold in plastic 

fire debris samples. DSC instruments are widely used for thermal characterization of 

polymers. With the help of DSC it is possible to reveal differences in the amount of 

cristallinity of the samples, which is due to the different manufacturing processes. 103 

Useful software for background matrices comparisons is available at the TWGFEX 

web site. The database is open access, which is why in a case of figuring out the 

matrix of a fire debris sample, the use of the site is recommended for screening the 

samples. 104 

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4 Modelling 

The importance of basics of modelling cannot be neglected and thus the same article 

by Olivier Delémont as in the previous review is mentioned here as well. 105 Fire 

safety studies were the first applications for modelling fires. Motivation for using 

modelling in forensic fire investigation has been occasional from the beginning of the 

1990s. Although it has been advised that all investigators should be aware of the 

meaning of modelling, it has been underused in forensic fire investigation. 106 The 

modelling technique has been divided into categories including hand calculations, 

zone models and field models. 105, 106 These are bases for different approaches 

according to the use, level of precision and complexity. Hand calculation uses basic 

mathematics and still provides a reliable prediction about the fire. The zone model 

uses distinguished layers, but the method suffers from consequent limitations due to 

the relative simplicity and the empirical assumption of stratification. 106 The field 

model, on the other hand, divides the fire environment into small cells and the fire 

fluid flows are calculated according to computational fluid dynamics (CFD) known as 

field models. Results of the field models are gained by using higher level 

mathematics and they include limitations due to some unavoidable errors and 

105, 106 

uncertainties. 

The most appropriate use of mathematical modelling in forensic fire investigation is in 

testing experiments of hypotheses made about the fire. Fire dynamic simulation (FDS) 

is software using computational fluid dynamics to provide evidence for fire 

investigation. It uses a collection of data obtained from the fire. This includes the 

samples, testimonies, chronology of the events and other matters that reduce the 

probability of hypotheses. Data obtained from FDS may be used by judicial 

authorities. For example, the data might prove claims made by an eye witness wrong 

or/and the data obtained by the model may bear visual significance for court. The 

model might also provide a timeline and analyse the progression of events. 

105, 107, 108 

Other circumstances for the use of modelling are situations in which the fire 

development was peculiar. In such cases, fire modelling may explain factors 

contributing to the fire and its specific propagation by using multiple hypotheses. 105, 

109 

A disadvantage of modelling is the time spent in the calculations, and thus the count 

of the mesh has to be reasonable but large enough to attain reliable results. Also 

diversity of construction materials in general creates a challenge for fire simulation. 

The method uses average values instead of accurate ones and therefore an error is 

106, 110, 111 

always present. 

Fire dynamic simulation has been used as a defined test plan for a full scale 

experimental. There the heat releasing rate (HRR) and temperature evolution were 

the most important features. 109 In other experiments, reconstruction of a hotel arson 

and fire growth in a tunnel were studied. The experiments provide important 

information not only to the forensics but also to the fire administrations. 107, 110, 112 A 

sealed structure of trains and a possibility of large fires due to arson have been a 

major influence to do numerical simulations of pressure changes in closed chambers. 

The prediction of pressure, temperature distribution and air flow pattern were studied. 

The experiment had a good correlation between prediction and experimental results. 

113 

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A novel model for prediction of a multi-component liquid being spilt for the purposeful 

intention of arson has been developed. The method presages concentrations of fuel 

vapours, evaporation rate and the flash point on a liquid fuel consisting of many 

components. The model uses Raoult’s law considering the activity coefficient and the 

estimated values corresponded well with the measured ones. 114 

One way model has been used to predict the ignition time and spontaneous ignition 

of some woods. The investigation has been both theoretical and experimental, 

consisting of pyrolysis and spontaneous ignition studies. The heat was radiated from 

upper electronic radiator and ignition was initiated without other external sources. 

Four different radiator powers and four tree species were tested. The modelling 

results matched well with the experimented ones. 115 

Different kinds of software of numerical simulation have been used mainly by the fire 

prevention purposes. For example, the selection of a fire origin room has been 

determined by a mathematical, multicriteria decision-making model in which fire 

loads are estimated. 116, 117 Also experimental investigations of fire spread and 

flashover time in office fires have been made. 118 Furthermore, the data obtained 

from the fire ignition location, probability of fire spread or evaluation of risks of fatality 

119, 120 

in building fires can all be calculated by the Bayesian network. 

The Bayesian theorem includes two separate but important features of forensic work: 

the question if scientific evidence can be based on a probabilistic argumentation and 

identification of forensic analyses. The method helps in sorting and evaluating 

analytical information in complex cases. 121 A few publications considering the 

Bayesian network have been published. The first considers the logic and 

argumentative implications in which examination is made with a distinct perspective 

in order to avoid false interpretations. 122 The second reviews the use of the methods 

in chemistry, including forensic sciences, chromatographic and mass spectrometric 

methods. 123 The third article serves as a guide in parameter estimation when using 

forensic data. It is the first part which introduces the way to apply this method; the 

second part is on its way and will be published this year. It will demonstrate the 

network fragments as building blocks while constructing larger interference 

models. 124 

The number of tragic accidents and fires in road tunnels during the past decades has 

been a starting point for tunnel safety and simulations of tunnel fire events. 125 

Several fire safety studies have been made considering fire scenarios in road tunnels. 

Many of the experiments and their results can be used in other tunnels, such as 

subway or river tunnels. 111 Here only a few of them are cited, in case of interest or a 

112, 126-132 

possible need for fire investigations. 

5. Educational aspects of fire investigation 

There are some problems in teaching fire investigation, such as a lack of 

uncontaminated fire fields, time dependence and varying circumstances. Thus the 

development of fire scene investigator students in understanding investigation 

theories, skills and applications has been studied in a virtual environment. A 

437

simulated environment in order to investigate the fire scene in a three dimensional 

model is produced by a software. The students got basic skills and their knowledge 

developed from the learning environment CD. A reward of the CD was to use the 

program when it suits best to the students’ schedule. The time the student could 

spend with the program was unlimited. On the other hand, limitations of the 

technique were that the field of view, visual acuity and navigation of the environment 

were limited. Although the results were promising, importance of real tasks in 

authentic fire places cannot be underestimated. 133 

Another point of view on arson investigation is the educational point for chemistry 

students. Using fictitious forensic cases, the studies connect the experiments to realworld 

applications. The sampling is made by absorbing the substances either by 

charcoal tubes or stripes, the identification and characterization of the accelerants is 

made by GC (FID) or GC-MS. The students have been able to develop their problemsolving 

abilities and their enthusiasm to understand chemistry has deepened. 134-137 

6. Explosions 

Explosions of flammable substances, such as natural gas or solid combustible dust, 

usually lead to casualties. Fires and explosions occur in many different industries, 

such as plastic products, chemical manufacturing and primary metal and wood 

factories. A large study considering the explosions during the last 25 years and an 

assessment about dust explosion hazards have been made. 138-140 

An assumption that materials are inert can lead to major incidents. This assumption 

underlies the explosion of aluminum mats when being restored. 141 Also in Italy a 

dust explosion occurred due to the domino effect caused by a spark inflaming a 

mixture of vegetable dust, wool fibres and inorganic substances. 142 Combustible 

dusts are a serious hazard in industrial manufacturing plants and multidisciplinary 

techniques might be needed to develop plausible theories for explosions and 

reconstructions. Such cases often include gases, for example acetylene, which burn 

very rapidly. 138, 143 A discussion of a BLEVE’s (Boiling Liquid Expanding Vapour 

Explosion) occurrence in a portable gasoline container has been attended. In the 

same abstract, it was shown that a flame arrester would have prevented the 

explosion inside of a gasoline container. 144 

Another matter is co-operation of forensics with the pathologist. A case report about 

an unusual suicidal explosive death reminds us of the importance to collaborate in 

order to find the correct cause of death. 139 

7. Criminological aspects of fire setting 

Arson and fire setting are significant concerns for public health and safety. The cost 

of arsons and their consequences are billions of dollars each year. On the other hand, 

there are damages paid for thousands of injuries and deaths annually. 145 During the 

last decades an enormous amount of global studies has been made to understand 

the behaviour of arsonists and firesetters. Lately the focus has been in juveniles and 

adolescents due to the opportunity to find a way to interfere in their behaviour at an 

early stage. 146-148 Thus persons under the age of 16 account for more than half of all 

fire-related arrests and the younger the persons were when setting the first fire, the 

438

more likely they are to become a frequent fire setter. 149 Furthermore, antisocial 

147, 150 

behaviour and alcohol abuse had more often relation with multiple fire settings. 

The link between maltreatment and anger disgorging with fire has been 

demonstrated. Fire setting seems to originate from difficulties in child’s emotions and 

behaviour. 151 Retrospective analyses of arsonists demonstrate that their intellectual 

disabilities (IQ lower than 85) 152 are more common than in general population; 

personality disorders and revenge serve often as reasons for fire setting. 153 Feelings 

of tension and excitement were more common in fire recidivism group. 145 Challenges 

in psychiatric and psychological research, such as the amount of population and 

classification of the surveys, are the contributors for not being able to find “a general 

145, 154 

arsonist”. 

A revealing article about pyro-terrorism and its potential to become a tactic of choice 

for terrorists has been published. Terrorist organizations are creative and try to 

balance the risk between gained attention and the costs of action. Thus low-tech 

methods have been applied when the target is highly vulnerable, meaning mainly the 

employment of arson. The article points out recent cases from Israel, Spain, Estonia 

and Greece. Furthermore, it reveals implications for Northern America. 155 

8. International co-operation 

8.1 Working groups and conferences 

International working groups aim to share knowledge, share experiences, collect 

information, develop methods and give advice to working group members. The 

European Network of Forensic Science (ENFSI) is recognized as an expert group in 

the field of forensic sciences. One of them is the Fire and Explosion Investigation 

Working group (FEIWG). The sphere of activities of the working group is field 

investigation, technical investigation and chemical analyses of fire debris samples. 

FEIWG strives to ensure the quality of development of fire and explosion 

investigation. There are three sub-committees: Training and Education, Accelerants 

and Explosion. The Working Group published a Practical Guide for First Responders 

to Fire Scene in March 2008. It has been translated into French, German, 

Portuguese and Spanish and will be translated into Swedish, Danish, Italian and 

Greek in 2010. FEIWG has organised two New Science Seminars in Switzerland. 

The first one in 2008 discussed Scene Investigation and in 2010 the topic was fire 

debris analysis. 156 

In the USA, the Technical Working Group of Fire and Explosion (TWGFEX) maintains 

co-operation among personnel in the Forensic laboratory, public safety, private 

investigation, and legal communities. The Working Group develops protocols and/or 

guides for collection and analysis of fire and explosion debris, training and quality 

assurance and introduces new techniques in the field of forensic fire and explosion 

investigation and laboratory analyses. TWGFE has three active scene committees: 

Education/Training, Fire Modeling Database and Scene Protocol Committee, and six 

laboratory committees: Explosives Education and Training, Explosives Database, 

Explosives Standard Protocols, Fire Education and Training, Ignitable Liquids 

Database and Fire Standard Protocols. TWGFEX has published a Training Outline 

for First Responders at Fire Scenes (October 2008) and Recommended Guidelines 

439

for Forensic Identification of Post-Blast Explosive Residues. TWGFEX organised a 

one-day symposium and many training courses in Orlando in 2009. 157 

The Ignitable Liquids Reference Collection (ILRC) was developed by the ILRC 

Committee of the TWGFEX. The Ignitable Liquids Reference Collection is a 

compilation of reference materials used by forensic analysts to conduct fire debris 

analysis. The ILRC consists of a comprehensive set of ignitable liquids and 

accompanying characterization data used in the analysis of fire debris samples in 

accordance with the American Society for Testing and Materials (ASTM) E-1618 

standard test methods. Another TWGFEX database is a substrate database, which is 

a tool designed for screening purposes only. It does not replace the need for 

obtaining comparison samples to evaluate the matrix, but it gives good tools for 

estimating the results. These two databases are easy to use and all information is 

valuable for regular work. 

Other important forensic discussion forums are international conferences. The 

European Academia of Forensic Science (EAFS) organises Triennial Meetings, 

stimulates OOS workshops and effective transfer of knowledge between institutions. 

In the EAFS conference organised in 2009, the working group (FEIWG) had two 

workshops: Fatal fires and Interpretation of analytical data. 156 The American 

Academia of Forensic Science (AAFS) organised annual meetings including 

workshops, presentations and posters. 158 The Australian and New Zealand Forensic 

Science Society (ANZFSS) holds an International Symposium every two years. The 

meeting and sessions cover the major disciplines of forensic science. 159 The 

International Association of Forensic Sciences (IAFS) brings together academics and 

practicing professionals of various disciplines in forensic sciences and organizes 

triennial meetings. 160 A part of Nordic co-operation are the meetings of the fire group 

(Nordisk brandgruppe). The group meets twice a year discussing the latest 

viewpoints of fire investigation. The meetings include seminars and some tests, such 

as interpretations of fire scene photographs. Also a manual for fire investigation was 

published in 1999 and it has been updated several times since then. 161 

8.2 International Collaborative Tests 

Quite many forensic laboratories are accredited and they need a collaborative or 

proficiency test to prove the reliability of the test results. One possibility is 

Collaborative Testing Services (CTS) 162 , which combines experience with forensic 

interlaboratory tests as Flammable Analyses. All participants report the results and 

methodology used in case it may help others to develop their own analyses. For the 

Flammables tests, examiners provide the detection, identification and comparison of 

flammable residue evidence. In 2009, the Flammables Analysis Test was sent to 371 

participants and 302 participants (81%) returned the data. Approximately 93% of the 

returners did the identification of the flammable substance correctly. 163 

The ENFSI Fire and Explosion Investigation Working Group is the provider of a 

Collaborative Testing Program for ignitable liquid analysis to Working Group 

members. The collaborative tests are designed to share and exchange knowledge on 

subjects such as techniques, products used as accelerants in various countries, 

matrix and weathering effects and so on. They are not designed to monitor the 

performance of individual laboratories like the proficiency tests. Participation in this 

440

ENFSI collaborative testing programme gives the laboratories an opportunity to 

review their original methods by taking advantage of the varied information derived 

from these exercises. The overall review of the laboratories’ results is carried out by 

the organising committee and represents one of its valuable outputs. 164 In the test 

organized in 2008, forty-five laboratories were listed to participate, eventually fortythree 

of them (96%) returned their analysis results for evaluation. From the 42 

laboratories that identified and classified the samples, 37 classifications were 

considered correct (88%). One laboratory was not able to detect this product in the 

165, 166 

sample. 

Table 1. Results of CTS-tests. 

CTS-test 2007 167 2008 168 2009 169 

Participants 329 345 371 

Returned 269 (82%) 286 (83%) 302 (81%) 

Correct answer from the returned 90-96% 94-98% 91-96% 

Table 2. Results of ENFSI/FEIWG-tests. 

ENFSI/FEIWG-test 2007 2008 2009 

Participants 41 45 44 

Returners 38 (93%) 43 (97%) 42 

varies per 

Correct answer from the returned 71-84% 37 (88%) element of the 

test 

9. Summary 

Although being perhaps the most important section of forensic fire investigation, 

sampling has not gained adequate attention in the studies. Samples gathered from 

fire scenes require new and suitable packaging methods. Packaging materials should 

be easy not only for carrying, but also for handling and sampling for different analysis 

techniques. The use of canine units is useful in field examinations and even their 

number should be raised. On the other hand, attention should be paid to the dogs’ 

training, in validation and monitoring their quality. 

Fuels have been classified e.g. on the basis of high temperature gas chromatography 

mass spectrometry (HTGC-MS) with a programmable temperature vaporizer injector 

and fast gas chromatography coupled with a fast scanning quadrupole ion trap mass 

spectrometer (GC-QIT MS). These methods were referred to in respect to fire debris 

analysis only in single articles. It will be interesting to see how these aforememtioned 

methods will be developed further. 

Statistical methods have gained some attention in analytics, but those applied for fire 

scene investigation will hopefully be developed in the near future. At the same time 

modelling may be a powerful tool for retrospectively establishing the cause and the 

starting point of fire. Applying modelling methods calls for deeper knowledge about 

441

information technology and this in fact may be one reason for modelling not being so 

widely spread in forensic casework. 

International co-operation will become more important because of increasing 

efficiency demands. Swift means of communication and common databases enable 

smooth co-operation within the forensic community. Despite modern IT technology, 

the demand for international conferences and personal discussions with scientists 

cannot be neglected. 

10. Books and other publications 

Robert D. Blackledge, Forensic Analysis on the Cutting Edge –New Methods for 

Trace Evidence Analysis, USA 2007, ISBN-10: 0-471-71644-8 

Carl Chasteen, Fire and Explosion Investigations and Forensic Analyses: Near- and 

Long-Term Needs Assessment for State and Local Law Enforcement, The National 

Center for Forensic Science, and The Thecnical/Scientific Working Group for Fire 

and Explosions, December 2008 

Oliver Pourret, Patric Naïm, Bruce Margot, Bayesian networks: A Practical Guide to 

Applications, John Wiley and Sons, Ltd., March 2008, ISBN 978-0-470-06030-8 

Christopher W. Schmidt, Steven A. Symes, The Analysis of Burned Human Remains, 

Elsevier Great Britain, 2008, ISBN 978-0-12-372510-3 

Eric Stauffer, Julia A. Dolan, Reta Newman, Fire Debris Analysis, Elsevier USA, 2008, 

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Craig L. Beyler, Analysis of the Fire Investigation Methods and Procedures Used in 

the Criminal Arson Cases Against Ernest Ray Willis and Cameron Todd Willingham, 

Texas Forensic Science Commission, Sam Houston State University College of 

Criminal Justice, 17 August 2009 

David Widlund, Elektricitet och bränder med inriktning på brandutredning, 

Myndigheten för samhällsskydd och beredskap, (Electricity and fire from fire cause 

determination viewpoint), Brandskyddsföreningen, elektriska nämnden Statens 

Kriminaltekniska Laboratorium, Karlstad, 2009, ISBN 978-91-7383-041-6 

Yao Lu, Forensic Applications of Gas Chromatography―Differential Mobility 

Spectrometry, Gas Chromatography/Mass Spectrometry, and Ion Mobility 

Spectrometry with Chemometric Analysis, Ph.D thesis, The University of Ohio, March 


Wolfram Jahn, Inverse Modelling to Forecast Enclosure Fire Dynamics, Ph.D thesis, 

The University of Edinburgh, 2010 

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452

Analysis and Detection of 

Explosives and Explosives Residues 


Richard A. Strobel 

Chief, Forensic Explosives Group 

United States Department of Justice 

Bureau of Alcohol, Tobacco, Firearms and Explosives 

Forensic Science Laboratory 

6000 Ammendale Road 

Ammendale, MD 20705 USA 

Greg Czarnopys 

Deputy Assistant Director Forensic Services 

United States Department of Justice 

Bureau of Alcohol, Tobacco, Firearms and Explosives 

Forensic Science Laboratory 

6000 Ammendale Road 

Ammendale, MD 20705 USA 

453


1.  INTRODUCTION AND COVERAGE OF THE LITERATURE  455  

2.  REVIEW ARTICLES  455  

3.  SAMPLING AND CONCENTRATION OF EXPLOSIVE TRACES  456  

4.  IDENTIFICATION OF EXPLOSIVES AND RESIDUES  457  

5.  INSTRUMENTAL ANALYSIS OF EXPLOSIVES  457  

5.1  LC/HPLC  457  

5.2  Ion Chromatography  458  

5.3  Capillary Electrophoresis  458  

5.4  Gas Chromatography  458  

5.5  Mass Spectroscopy  458  

5.6  Isotope Ratio Mass Spectroscopy, IRMS  460  

5.7  General Spectroscopy: Fluorescence, Luminescence, Spectrophotometric, UV, Chemiluminescence 460  

5.8  FTIR  460  

5.9  Raman Spectroscopy  461  

5.10  DSC, Thermal Analysis, TG  461  

5.11  Medical; Pathology; Injuries; Toxicology  461  

6.  DETECTION  461  

7.  ENVIRONMENTAL  461  

8.  ACKNOWLEDGEMENTS  462  


454

1. Introduction and Coverage of the Literature 

The coverage of the literature in this review period is a beneficiary (or victim) of the 

improved computer networking, the web, and the growth of abstracting services utilized 

by library technicians worldwide. Hence the record-setting number of citations listed. 

The additional citations are not the result of an increase in research in the forensic field 

but are the result of the ability to survey many more specialty journals which were 

previously unknown to the forensic practitioner. Without the current technology which 

allows one to easily view the abstracts in these far afield journals, it would be 

impossible in either expense or time to research them. It has been recognized for 

some time that the science in forensic science is largely derivative of more basic 

research and methods development activities. Forensic science methods are more 

accurately described as applications of existing science and technology. This 

demonstrates the requirement for the forensic scientist to be familiar with the literature 

of other scientific fields in order to advance. This is the reason that many citations are 

included as they might provide the spark of an idea for a forensic application. The two 

fields which greatly add to the volume of citations are the fields of environmental 

science and explosives detection technologies which will undoubtedly contribute to 

future advancements in forensic techniques. 

With nearly 600 references this author and anyone plowing their way through those 

would have a difficult time evaluating the usefulness of the material. Because of this 

the hyperlinks to abstracts of the articles are included. One can usually get an idea 

whether to pursue the article through a quick review of the abstract. Another feature 

which the reader should make use of is the “bookmark” feature in Microsoft Word. This 

will greatly assist navigation through the bibliography section starting on page 11 of this 

document. With the volume of references, the reader will have to excuse the fact that 

the author would never have sufficient time to comment on every article. Very valuable 

resources are any review articles which usually are specific to subject areas of interest 

and encompass references for numerous time periods. These will be the first articles 

discussed. 

2. Review Articles 

An all encompassing review is provided by Brettell, [1] et al on Forensic Science with a 

subject section devoted specifically to explosives. The period of coverage is pre-2007. 

A review article specifically dedicated to explosives analysis appeared in the Journal of 

Analytical and Bioanalytical Chemistry by Moore and Goodpaster [2] covering the 

period up till 2009. In the field of detection Singh Suman [3] addressees approaches to 

explosives detection in a June 2007 article. Instrumental specific review articles include 

an article covering HPLC analysis of explosives by Gaurav [43] in the journal Critical 

Reviews in Analytical Chemistry in 2007. Continuing with HPLC analysis Fedorowski 

and LaCourse review post-column photochemical systems with electrochemical 

detection [47]. In the medical field and more specifically in battlefield injuries, Ulrich, 

Kalbermatten, et al, explore the possibility cancer caused by grenade fragments [152] 

via a literature review. The environmental field Viada, et al review explosive residue 

455

effects on sea turtles in a review [319]. Ghafari, et al explore removal techniques for 

nitrates in wastewater [386]. 

The explosives detection field includes a review of biosensor technology in a paper by 

Smith, D’Souza, et al [161] covering biosensor and biologically inspired explosives 

detection systems. Laser-based standoff detection of explosives is reviewed by Wallin, 

et al [164]. Explosives detection via Laser-Induced Breakdown Spectroscopy, LIBS, is 

examined by Gottfried, et al [165] in a review article covering advances, challenges, 

and future advances in a 2009 article in the journal Analytical and Bioanalytical 

Chemistry. It is important to note the possible forensic applications of this technique in 

the detection of post-blast explosive residues. In the Spectroscopic area Neutron 

Activation Analysis receives a renewed look in a review in Spectroscopy Reviews in an 

article by Hee-Jung [174]. In the area of air cargo explosives detection Runkle, et al 

[215] looks a Photon and Neutron techniques for detection in a review. The FAIMS 

technique utilizing IMS and differential IMS is explored by Kolakowski [241] in the 

journal Analyst. 

3. Sampling and Concentration of Explosive Traces 

Whether involving recovery of traces of explosives for post-blast identification or 

similarly small amounts for detection of explosives in security applications a critical step 

involves selectively trapping and separating the explosive molecule from every other 

chemical compound present. A number of techniques are examined such as Zeichner, 

et al studying the use of adhesive coated stubs for collection of explosives residues 

from a suspect’s hands [7]. Another technique for collection of residues from humans 

is Oxley, et al, examining factors for sorption and collection of explosives from hair [13]. 

Solid phase extraction is frequently examined both for collection and clean-up of 

explosive traces. Articles studying the application of SPME (Solid Phase Micro- 

Extraction) appeared in the Journal of Chromatography where Harvey, et al looked 

SPME use with specially coated fibers [8]. Metal β-diketonate polymers were utilized 

to capture explosives from vapor in a paper by Harvey [9]. The continued use of 

SPME for sample collection and clean-up has made a review of the Solid Phase 

Sorbents a necessity. This is accomplished in a review article on the subject by 

Tachon, et al [11] which examines the use of sorbents for clean-up of organic 

explosives. A current explosive of interest, urea nitrate, is covered in two articles by 

Almog, et al [12] and Tamiri [40]. The Tamiri article addresses initial studies while 

describing the synthesis and the Almog article covers trace recovery and detection of 

urea nitrate. 

The success or failure to identify explosive traces begins at the scene of the crime. 

Failure here makes subsequent laboratory analysis fruitless. If a forensic scientist can 

insure quality samples are collected at the scene through preliminary testing 

subsequent laboratory analysis will have a higher probability of success. The 

Netherlands Forensic Institute examined the scene detection of explosives in a study 

by van Asten [17] where the technology of Raman Spectroscopy and GC-MS with 

SPME sampling at an explosives scene is examined. Maximizing the amount of 

456

evidence gathered at a bombing scene is always crucial to the successful outcome of 

an investigation. The key is found not only in the collection but also preservation of 

evidence. Bille [16] examines on-site cyanoacrylate fuming to preserve fingerprint 

evidence and the subsequent effect on both DNA and explosive residue examination. 

4. Identification of Explosives and Residues 

Major interest remains in all types of peroxide explosives whether for detection pre- 

and post-blast. Also of interest are the properties of these explosives. Detection for 

security reasons is addressed in papers by Burks [18] which examines current trends; 

Benedet [19] which examines amperometric sensing; Lock [20] which using specific 

films for specific microcantilever detectors; Banerjee [21] examining nanotube sensor 

technology; Bohrer [22] looking a vapor phase detection and Cotte-Rodriguez [23] 

examining in-situ detection by AP-CI-MS with sampling via desorption electrospray 

ionization. The properties of peroxide explosives, specifically TATP, are provided in a 

2-part paper by Zeman [25], [26] which examines TATP mixtures. Oxley [28] examines 

Raman and Infrared spectroscopy of peroxide explosives. Detection of HMTD is 

examined in papers by Laine[29], Persons [30] and Parajuli [31]. Detection of TATP by 

a wide variety of techniques is examined in papers by Shen [32], Xie [33], Wilkinson 

[34], Räsänen [35], Lubczyk [36], Capua [37], Bauer [38] and Germain [39]. The 

papers cover a complete variety of instrumental techniques from fluorescence, mass 

spectroscopy, custom sensors, terahertz spectroscopy to chronoamperometry. 

A selection of papers covering novel explosives and properties, degradation of 

explosives and performance characteristics are included in references [419] to [546]. It 

is beyond the scope of a forensic review article to cover these in depth but they are 

provided to the reader for convenient further investigation. 

Explosive residues are the primary evidentiary examined by the forensic scientist. Two 

articles covered issues directly related to the subject: Vermeij [41] looked specifically at 

the morphology and the chemical composition of pyrotechnic residues as affected by 

the degree of confinement at the time of explosives. Kunz [42] measured amounts of 

explosives residues in trace situations for the purpose of guiding future development of 

sensors for pre-blast detection. 

5. Instrumental Analysis of Explosives 

5.1 LC/HPLC 

Much of the work of the forensic scientist in the laboratory is devoted to utilization of 

instrumental techniques to identify explosive traces. LC or HPLC continues to be a vital 

separation technique and when coupled with a specific detection method, an 

identification technique. The review article by Gaurav [43], mentioned previously, 

provides a good overview of LC techniques as applied to explosives. Many of the 

articles in this section deal with specific applications for solving specific problems in 

explosives analysis with many covering explosives processing issues; Gaurav [44], 

457

Bausinger [45]. Borowsky [46] looks at development of an electroosmotic pump for 

development of LC on a microchip moving forward to the goal of lab-on-a-chip 

technology for explosives detection. In order to further the selectivity of detection 

systems for use with LC, Fedorowski reviews post-column photochemical reaction 

systems [47] coupled with electrochemical detection. Gaurav [48] examines a SPME 

followed by LC-UV analysis for explosives in aqueous samples. 

5.2 Ion Chromatography 

The technique of Ion Chromatography has received an upgraded role in forensic postblast 

analysis with the addition of a more specific detector, the mass spectrometer. 

The following two articles utilized conductivity detection but the separation techniques 

prove valuable. Johns [50] offers a system that identifies 18 anions and 12 cations of 

interest for post-blast analysis. Additionally the utility of the technique was tested with 

the actual post-blast analysis of witness plate materials and soil. The results were 

confirmed using capillary electrophoresis. Meng [49] tests five inorganic anions and 

three cations in a single chromatographic run using a non-suppressed ion 

chromatographic technique. Lang and Boyle [51] apply IC-MS to identify black powder 

substitutes. 

5.3 Capillary Electrophoresis 

Tagliaro[54] offers a review article on recent advances in the application of CE to 

Forensic Science with a section specifically devoted to explosives. Other articles by 

Piccin [52], Pumera [53][55] examine electrophoresis applied on a microchip for 

explosives detection. Hutchinson [56] uses a portable CE to examine post-blast 

inorganic anions and cations. Indirect photometric detection was accomplished by 

utilizing a miniature LED detector. Fifteen anions and 12 cations of interest in 

explosives analysis were separated and detected. 

5.4 Gas Chromatography 

GC continues to play a role in the separation of explosive compounds from various 

matrices prior to detection by a variety of means. Li [57] offers a SPME technique 

coupled with GC for determination of the DMNB tagging agent for plastic explosives. 

Burleson [58] examines gunpowder residue by using SPME extraction and GC coupled 

with a nitrogen-phosphorous detector. Nitroaromatic compounds and their Kovats 

Retention Indices are examined in Caulfield [59]. A novel capillary column coating is 

offered by Harvey [60] for selective retention of explosives related compounds. 

5.5 Mass Spectroscopy 

The variety of mass spectrometric techniques has blossomed in recent years. 

Options are numerous in the selection of type of mass detector and the initial means 

for fragmenting the molecule of interest. Bečanová [61] examined TNT and the 

degradation of TNT with LC-electrospray ionization MS. Nitroaromatic explosive 

458

compounds were also examined by Adeem [73], McEnnis [81] using laser induced 

fragmentation and Sulzer [82] who examined dinitrobenzene, a toxic yet very 

insensitive explosive. Alizadeh [62] utilized electron attachment to nitroaromatic 

compound 2-nitro-m-xylene in the gas phase. Schramm [63] used thermal 

desorption single photon ionization ion-trap MS (TD-SPI-ITMS) in an application for 

real-time detection of explosives for screening wipes. The technique offers very 

gentle ionization to maintain the molecule’s mass and detection selectivity. In 

addition a successful field trial is included as demonstration the technique’s 

applicability to security screening. Scherperel [64] examines smokeless powders 

with nanoelectrospray ionization MS (nESI-MS). The technique provides forensic 

identification of residues by examining the organic stabilizers to identify and 

differentiate unburned smokeless powders from different brands of ammunition in 

gunshot residue analysis. Chen [65] looked at utilizing EESI-MS for examination of 

explosive traces on human skin. Mullen [66] used TOF-MS to identify 

nitroaromatics and related compounds using laser photoionization. Martinez-Lozano 

[67] used secondary electrospray ionization (SESI) to identify explosives in the vapor 

state at PPT concentrations. Collin [68] used two techniques, GC separation with 

Quadrapole Ion Trap (QIT) and a second technique with fast GC separation followed 

by dynamic collision-induced dissociation (DCID). Picogram sensitivity is claimed, 

rapid analysis (2.5 second GC runs) and the two MS techniques used together are 

offered as a confirmational technique. Song [69] compares NI-DART with NI-APPI 

for a wide range of compounds including explosives. Both techniques offer 

numerous ionization mechanisms for a wider range of compound identification. ESI 

and APCI techniques are compared and contrasted. Mauracher [70] examined 

musk-ketone compounds with the present results compared with previous aromatic 

nitrocompounds studied in our laboratory recently. Particularly the close similarity of 

musk ketone and the explosive trinitrotoluene is of special interest. Talaty [71] 

analyzed for explosives and on fabric samples by ambient DESI. Sampling was 

done in-situ without sample preparation and with a wide variety of common 

interferent compounds. High sample throughput and picogram sensitivity are 

reported for a technique which could be used for security screening. Wells [72] used 

field portable MS instrumentation utilizing DART and DESI ionization. The team 

used a portable MS with API interface adapted to DESI and DART sample 

introduction to rapidly analyze explosives and other compounds of security interest. 

Huestis [74] and Schramm [75] measured ionization potentials for a number of 

compounds including nine explosives for investigating whether the compounds could 

potentially be detected via single photon ionization time of flight mass spectrometry 

(SPI-TOFMS). Na [76] used an ambient ion source to detect explosives on solid 

surfaces. Justes [77] used a similar source to examine explosives on skin. 

Williamson [78] provides a review which covers the rationale for using TOF for LC 

detection, and describes the many methods currently in the literature for the 

quantitation of pharmaceuticals, environmental pollutants, explosives and many 

phytochemicals. Jönsson [79] used SPME sample collection with (GC–ECNI-MS) 

and isotope dilution quantification for the analysis of nitroaromatic compounds in 

complex, water based samples. Ochsenbein [80] compares SPME and direct 

injection techniques for explosive compounds in water. 

459

5.6 Isotope Ratio Mass Spectroscopy, IRMS 

IRMS continues to stimulate interest in the field of explosives analysis. Its full 

potential has yet to be realized but some specific applications are represented 

among the following publications. Lock [84] [92] looks at isotope linkages between 

precursor and high explosive products using IRMS. Quirk [83] looks at IRMS 

characteristics of plastics in post-blast debris. Sources of explosives are examined 

using IRMS in a paper by Widory [85]. Mechanisms for explaining detonation 

mechanisms are studied in papers by Anisichkin [89] and [90]. Environmental 

analysis using IRMS are outlined in papers by Bernstein [86], [88] and Amaral [87]. 

Specific explosives are examined by Gentile [91], ammonium nitrate and black 

powder; Benson [93], ammonium nitrate and Benson [94], TATP and PETN. Results 

of four inter-laboratory comparisons in a test provided by the FIRMS network are 

discussed by Carter [95]. Proceedings from the FIRMS network Conference in 2005 

are provided by Wakelin [96]. 

5.7 General Spectroscopy: Fluorescence, Luminescence, 

Spectrophotometric, UV, Chemiluminescence 

Many of the techniques offered in the spectroscopy area fall into distinct categories 

of spectrum. Many of the techniques apply to specific explosive compounds or 

processes. Some of the techniques are applied to detection as opposed to 

applications for the forensic laboratory. Never the less they could have a future 

application. Spectrophotometric techniques are offered by Üzer [99] for 

determination of RDX in mixes and residues. A selective spectrophotometric 

technique for TNT is covered by Erçağ [102] using a colorimetric sensor. 

Fluorescence based techniques are presented by Sanchez [106] for nitroaromatics, 

nitramine and nitrate ester explosive residues. Fluorescence quenching based 

sensors and techniques are offered by Shi [107], Germain [108], Zyryanov [110], 

Monterola [113], Sheaf [114], Meaney [116], Hughes [117], Sushobhan [118], Naddo 

[111] and Andrew [120]. Techniques for use in stand-off detection of explosives are 

covered by Babushok [100], Pacheco-Londoño [101] and Nadezhdinskii [104]. 

Wang [97] examines laser induced plasma for plastics and an explosive. Ramos 

[98] looks at photochemistry and its use with cavity ringdown spectroscopy. A 

specific spectroscopic sensor for TNT is explored by Changmin Deng [103]. UV 

plays a role in Chaudhary’s [105] work which looks at TNT and RDX detection and 

Sharma [109] who uses UV to provide photo-fragmentation detection for TNT. 

Meaney [112] provides a review of Luminescence based detection systems and 

Sanchez [119] looks at explosives sensing with 1,1-Tetraphenylsilole- and 1,1- 

Silafluorene-vinylene Polymers through their luminescence properties. Naddo[121] 

uses a novel fluorescent nanofibril film for explosives detection. 

5.8 FTIR 

Fourier Transform Infrared Spectroscopy has had a long tenure in forensic 

explosives analysis. The technique continues this use in Banas [122] paper on post- 

460

last traces of explosives and Mou’s [124] work on ATR-FTIR use for identification of 

explosive particles in fingerprints. Sharmaand [123] examines EDA complexes with 

amines for determination of vertical electron affinity of TNT. 

5.9 Raman Spectroscopy 

The single largest application of the Raman technique is in stand-off detection of 

explosives. Papers covering this aspect are offered by Ramos [125], Hobro [126], 

Gaft [132], Katz [134], Li [135] and Eliasson [136]. The last reference covering the 

more recent interest in detection techniques for liquid explosives. Raman has also 

proved useful for in-situ analysis of explosives such as in fingerprints, Emmons [127] 

and Esam [129] utilizing a confocal Raman microscope to look at explosive particles 

on clothing. Portnov [131] uses backward coherent anti-stokes Raman to detect 

explosive particles. Fleger [128] examined narrow gated Raman and the 

luminescence of explosives. Nagli [133] looked at UN, PETN, TATP, RDX, TNT and 

PETN and determined the Raman cross-section dependence on excitation energy in 

spectral range 620–248 nm. 

5.10 DSC, Thermal Analysis, TG 

Korošec [137] examined emulsion explosives and utilized both TG and DSC for 

quantitative determination of the individual nitrates and water in ‘water in oil’ 

emulsion explosives. 

5.11 Medical; Pathology; Injuries; Toxicology 

A variety of articles covering medical aspects of explosives are provided in references 

138-152. 

6. Detection 

References for explosives detection are numerous and generally beyond the scope of 

the forensic analysis of explosives. The number are undoubtedly due to monetary 

investment in explosives detection technology for security purposes. The fact remains 

that many of the detection technologies might ultimately find a use in the forensic 

laboratory and are included because of this. IMS is but one prominent example of this. 

For the reader the ”Detection” section is broken down according to specific detection 

technologies. The section is arranged as follows: Canine Explosives Detection, 

Biosensors, LIBS Detection, Neutron, Terahertz, Nuclear Techniques (NQR, 

Quadrupole Resonance), X-Ray, Ion Mobility Spectroscopy, Human Factors, Novel 

Detection. 

7. Environmental 

Another major contributor to forensic techniques are analytical techniques 

specifically tailored to environmental analysis of explosives. Environmental 

461

equirements mandate the monitoring of explosive compounds and by-products 

during the manufacturing process and later in the environment at large. References 

in this section are arranged as follows: Environmental (general), Soil, Water and 

Wastewater, Bioremediation and Biodegredation. 

8. Acknowledgements 

Ms. Susan Barned, Librarian for the Bureau of Alcohol, Tobacco and Firearms 

Laboratory obtained virtually all of the literature citations and journal articles that 

were necessary to prepare this survey report. Her many hours of work are greatly 

appreciated. 

9. References 

Review Articles 

[1] Brettell, T. A.; Butler, J. M.; Almirall, J. R Forensic Science. Analytical 

Chemistry, 6/15/2007, Vol. 79 Issue 12, p4365-438 

  

[2] Moore, David S.; Goodpaster, John V Explosives analysis. Analytical & 

Bioanalytical Chemistry, Sep2009, Vol. 395 Issue 2, p245-246 

  

[3] Suman Singh. Sensors -- An effective approach for the detection of explosives 

Journal of Hazardous Materials 144.1-2 (June 1, 2007): p.15(14). 

  

Explosives Standards Reference Material, Laboratory Quality Control, 

Contamination Prevention 

[4] MacCrehan, William A. A NIST Standard Reference Material (SRM) to Support 

the Detection of Trace Explosives. Analytical Chemistry, 9/1/2009, Vol. 81 

Issue 17, p7189-7196 

[5] Robert A. Fletcher, Jacquelyn A. Brazin, Matthew E. Staymates, Bruce A. 

Benner Jr. and J. Greg Gillen. Fabrication of polymer microsphere particle 

standards containing trace explosives using an oil/water emulsion solvent 

extraction piezoelectric printing process Talanta Volume 76, Issue 4, 15 

August 2008, Pages 949-955 

[6] Matthew S. Beardah, Sean P. Doyle and Charlotte E. Hendey. Effectiveness of 

contamination prevention procedures in a trace explosives laboratory Science 

& Justice 47.3 (Nov 2007): p.120-124.  

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Sample Preparation, Sample Recovery, Sample Clean-up, Sample 

Preconcentration 

[7] Arie Zeichner, Sara Abramovich-Bar, Tsippy Tamiri and Joseph Almog. A 

feasibility study on the use of double-sided adhesive coated stubs for sampling 

of explosive traces from hands Forensic Science International Volume 184, 

Issues 1-3, 30 January 2009, Pages 42-46 

[8] Scott D. Harvey Selective solid-phase microextraction of explosives using 

fibers coated with the La(III) complex of p-di(4,4,5,5,6,6,6-heptafluoro-1,3hexanedionyl)benzene 

Journal of Chromatography A Volume 1213, Issue 2, 

12 December 2008, Pages 110-117 

[9] Scott D. Harvey and Thomas J. Wenzel (2008) Selective gas-phase capture of 

explosives on metal β-diketonate polymers Journal of Chromatography A 

Volume 1192, Issue 2, Pages 212-217 (May 30) 

[10] Ioana Voiculescu, Mona Zaghloul and Nachchinarkkinian Narasimhan (2008) 

Microfabricated chemical preconcentrators for gas-phase microanalytical 

detection systems TrAC Trends in Analytical Chemistry Volume 27, Issue 4, 

Pages 327-343 

[11] Romain Tachon, Valérie Pichon, Martine Barbe Le Borgne and Jean-Jacques 

Minet (2008) Comparison of solid-phase extraction sorbents for sample cleanup 

in the analysis of organic explosives Journal of Chromatography A Volume 

1185, Issue 1, Pages 1-8 

[12] Almog, Joseph; Burda, Gila; Shloosh, Yael; Abramovich-Bar, Sara; Wolf, Ehud; 

Tamiri, Tsippy Recovery and Detection of Urea Nitrate in Traces. Journal of 

Forensic Sciences, Nov2007, Vol. 52 Issue 6, p1284-129 

[13] Oxley JC, Smith JL, Kirschenbaum LJ, Marimganti S, Accumulation of 

explosives in hair--part II: factors affecting sorption. Journal of Forensic 

Sciences [J Forensic Sci], ISSN: 0022-1198, 2007 Nov; Vol. 52 (6), pp. 1291-6; 

  

[14] Michael R. Walsh, Marianne E. Walsh and Alan D. Hewitt Energetic residues 

from field disposal of gun propellants Journal of Hazardous Materials Volume 

173, Issues 1-3, 15 January 2010, Pages 115-122 

[15] Üzer, Ayşem; Erçağ, Erol; Apak, Reşat Selective colorimetric determination of 

TNT partitioned between an alkaline solution and a strongly basic Dowex 1-X8 

anion exchanger. Forensic Science International, Jan2008, Vol. 174 Issue 2/3, 

p239-243, 

  

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Crime Scene: Field Detection 

[16] Bille TW, Cromartie C, Farr M. Effects of cyanoacrylate fuming, time after 

recovery, and location of biological material on the recovery and analysis of 

DNA from post-blast pipe bomb fragments J Forensic Sci. 2009 

Sep;54(5):1059-67 

[17] A.C. van Asten, J. Dalmolen, M. Koeberg, A. Schoina and T. Pacqué Noninvasive 

detection of explosives at the crime scene Science &Justice Volume 

50, Issue 1, March 2010, Page 35 

  

Hydrogen Peroxide, Peroxides, General 

[18] Burks, Raychelle M.; Hage, David S. Current trends in the detection of 

peroxide-based explosives. Analytical & Bioanalytical Chemistry, Sep2009, 

Vol. 395 Issue 2, p301-313 

  

[19] Benedet, John; Donglai Lu; Cizek, Karel; Belle, Jeff La; Wang, Joseph. 

Amperometric sensing of hydrogen peroxide vapor for security screening. 

Analytical & Bioanalytical Chemistry, Sep2009, Vol. 395 Issue 2, p371-376 

  

[20] John P. Lock, Edward Geraghty, Lawino C. Kagumba and Ken K. Mahmud 

Trace detection of peroxides using a microcantilever detector Thin Solid Films 

Volume 517, Issue 12, 30 April 2009, Pages 3584-3587 

[21] Subarna Banerjee, Susanta K Mohapatra, Mano Misra and Indu B Mishra The 

detection of improvised nonmilitary peroxide based explosives using a titania 

nanotube array sensor Nanotechnology 20:7 p. 075502 

[22] Bohrer FI; Colesniuc CN; Park J; Schuller IK; Kummel AC; Trogler WC 

Selective detection of vapor phase hydrogen peroxide with phthalocyanine 

chemiresistors Journal Of The American Chemical Society [J Am Chem Soc] 

2008 Mar 26; Vol. 130 (12), pp. 3712-3 

[23] Ismael Cotte-Rodriguez, Heriberto Hernandez-Soto, Hao Chen and R. Graham 

Cooks. In-situ trace detection of peroxide explosives by desorption electrospray 

ionization and desorption atmospheric pressure chemical ionization. Analytical 

Chemistry 80.5 (March 1, 2008): p.1512(8). 

  

[24] Luciano Carlos, Debora Fabbri, Alberto L. Capparelli, Alessandra Bianco 

Prevot, Edmondo Pramauro and Fernando García Einschlag Effect of 

simulated solar light on the autocatalytic degradation of nitrobenzene using 

Fe 3+ and hydrogen peroxide Journal of Photochemistry and Photobiology A: 

Chemistry Volume 201, Issue 1, 1 January 2009, Pages 32-38 

464

[25] Svatopluk Zeman, Waldemar A. Trzciński and Robert Matyáš (2008) Some 

properties of explosive mixtures containing peroxides Part I. Relative 

performance and detonation of mixtures with triacetone triperoxide Journal of 

Hazardous Materials Volume 154, Issues 1-3 Pages 192-198 

[26] Svatopluk Zeman and Cécile Bartei (2008) Some properties of explosive 

mixtures containing peroxides Part II. Relationships between detonation 

parameters and thermal reactivity of the mixtures with triacetone triperoxide. 

Journal of Hazardous Materials Volume 154, Issues 1-3, Pages 199-203 

[27] Ming-Jer Liou and Ming-Chun Lu (2008) Catalytic degradation of explosives 

with goethite and hydrogen peroxide Journal of Hazardous Materials Volume 

151, Issues 2-3, Pages 540-546 

[28] Oxley J, Smith J, Brady J, Dubnikova F, Kosloff R, Zeiri L, Zeiri Y Raman and 

infrared fingerprint spectroscopy of peroxide-based explosives. Appl 

Spectrosc. 2008 Aug;62(8):906-15. 

HMTD 

[29] Derek F. Laine and I. Francis Cheng Electrochemical detection of the 

explosive, hexamethylene triperoxide diamine (HMTD) Microchemical Journal 

Volume 91, Issue 1, January 2009, Pages 125-128 

[30] Persons J, Harbison GS (2007) The (14)N quadrupole coupling in 

hexamethylene triperoxide diamine (HMTD Magn Reson Chem. 45(11):905-8 

  

[31] Parajuli, Suman; Wujian Miao Sensitive Determination of Hexamethylene 

Triperoxide Diamine Explosives, Using Electrogenerated Chemiluminescence 

Enhanced by Silver Nitrate. Analytical Chemistry, 7/1/2009, Vol. 81 Issue 13, 

p5267-5272 

  

TATP 

[32] Chengyin Shen, Jianquan Li, Haiyan Han, Hongmei Wang, Haihe Jiang and 

Yannan Chu Triacetone triperoxide detection using low reduced-field proton 

transfer reaction mass spectrometer International Journal of Mass 

Spectrometry Volume 285, Issues 1-2, 1 August 2009, Pages 100-103 

[33] Yuqun Xie and I. Francis Cheng. Selective and rapid detection of triacetone 

triperoxide by double-step chronoamperometry. Microchemical Journal Volume 

94, Issue 2, March 2010, Pages 166-170 

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[34] John Wilkinson , , Christopher T. Konek, Jesse S. Moran, Ewelina M. Witko and 

Timothy M. Korter. Terahertz absorption spectrum of triacetone triperoxide 

(TATP). Chemical Physics Letters Volume 478, Issues 4-6, 27 August 2009, 

Pages 172-174 

[35] Riikka-Marjaana Räsänen, Marjaana Nousiainen, Kaleva Peräkorpi, Mika 

Sillanpää, Lauri Polari, Osmo Anttalainen and Mikko Utriainen Determination of 

gas phase triacetone triperoxide with aspiration ion mobility spectrometry and 

gas chromatography–mass spectrometry. Analytica Chimica Acta Volume 623, 

Issue 1, 8 August 2008, Pages 59-65 

[36] Daniel Lubczyk, Carsten Siering, Jürgen Lörgen, Zinaida B. Shifrina, Klaus 

Müllen and Siegfried R. Waldvogel. Simple and sensitive online detection of 

triacetone triperoxide explosive. Sensors and Actuators B: Chemical Volume 

143, Issue 2, 7 January 2010, Pages 561-566 

[37] Eyal Capua, Roberto Cao, Chaim N. Sukenik and Ron Naaman. Detection of 

triacetone triperoxide (TATP) with an array of sensors based on non-specific 

interactions. Sensors and Actuators B: Chemical Volume 140, Issue 1, 18 June 

2009, Pages 122-127 

[38] C Bauer, U Willer, R Lewicki, A Pohlkötter, A Kosterev, D Kosynkin, F K Tittel 

and W Schade. A Mid-infrared QEPAS sensor device for TATP detection. 

Journal of Physics: Conference Series Volume 157, Number 1 012002 

[39] Germain ME, Knapp MJ. Turn-on fluorescence detection of H2O2 and TATP. 

Inorg Chem. 2008 Nov 3;47(21):9748-50 

Urea Nitrate 

[40] Tamiri, Tsippy; Rozin, Rinat; Lemberger, Nitay; Almog, Joseph. Urea nitrate, 

an exceptionally easy-to-make improvised explosive: studies towards trace 

characterization. Analytical & Bioanalytical Chemistry, Sep2009, Vol. 395 

Issue 2, p421-428 

  

Residues and Residue Characerization 

[41] Erwin Vermeij Willem Duvalois, Rutger Webb and Mattijs Koeberg 

Morphology and composition of pyrotechnic residues formed at different levels 

of confinement Forensic Science International Volume 186, Issues 1-3, 15 

April 2009, Pages 68-74 

466

[42] Kunz, Roderick R.; Gregory, Kerin Clow; Hardy, Dennis; Oyler, Jonathan; 

Ostazeski, Stanley A.; Fountain III, Augustus Way Measurement of trace 

explosive residues in a surrogate operational environment: implications for 

tactical use of chemical sensing in C-IED operations. Analytical & Bioanalytical 

Chemistry, Sep2009, Vol. 395 Issue 2, p357-369 

  

LC; HPLC 

[43] Gaurav, Dhingra; Malik, Ashok Kumar; Rai, P. K High-Performance Liquid 

Chromatographic Methods for the Analysis of Explosives. Critical Reviews in 

Analytical Chemistry, 2007, Vol. 37 Issue 4, p227-268 

  

[44] Gaurav, Malik AK, Rai PK Solid phase microextraction-high performance liquid 

chromatographic determination of octahydro-1,3,5,7-tetranitro-1,3,5,7tetrazocine 

(HMX) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in the 

presence of sodium dodecyl sulfate surfactant. J Sep Sci. 2008 

Jul;31(12):2173-81 

[45] Tobias Bausinger and Johannes Preuss Stability of nitroaromatic specialty 

explosives in reversed-phase liquid chromatographic systems Journal of 

Hazardous Materials Volume 162, Issues 2-3, 15 March 2009, Pages 1578- 

1582 

[46] Borowsky JF, Giordano BC, Lu Q, Terray A, Collins GE. Electroosmotic flowbased 

pump for liquid chromatography on a planar microchip. Anal Chem. 

2008 Nov 1;80(21):8287-92 

[47] Jennifer Fedorowski and William R. LaCourse A review of post-column 

photochemical reaction systems coupled to electrochemical detection in HPLC 

Analytica Chimica Acta Volume 657, Issue 1, 4 January 2010, Pages 1-8 

[48] Gaurav, Ashok Kumar Malik and P.K. Rai Development of a new SPME– 

HPLC–UV method for the analysis of nitro explosives on reverse phase amide 

column and application to analysis of aqueous samples Journal of Hazardous 

Materials Volume 172, Issues 2-3, 30 December 2009, Pages 1652-1658 

Ion Chromatography 

[49] Hong-Bo Meng, Tian-Ran Wang, Bao-Yuan Guo, Yuki Hashi, Can-Xiong Guo 

and Jin-Ming Lin Simultaneous determination of inorganic anions and cations 

in explosive residues by ion chromatography Talanta Volume 76, Issue 2, 15 

July 2008, Pages 241-245 

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[50] Cameron Johns, Robert A. Shellie, Oscar G. Potter, John W. O’Reilly, Joseph 

P. Hutchinson, Rosanne M. Guijt, Michael C. Breadmore, Emily F. Hilder, Greg 

W. Dicinoski and Paul R. Haddad (2008) Identification of homemade inorganic 

explosives by ion chromatographic analysis of post-blast residues Journal of 

Chromatography A Volume 1182, Issue 2, Pages 205-214 (Feb 29) 

[51] Lang GH, Boyle KM. The analysis of black powder substitutes containing 

ascorbic acid by ion chromatography/mass spectrometry. J Forensic Sci. 2009 

Nov; 54(6):1315-22 

Capillary Electrophoresis 

[52] Piccin E, Dossi N, Cagan A, Carrilho E, Wang J. Rapid and sensitive 

measurements of nitrate ester explosives using microchip electrophoresis with 

electrochemical detection. Analyst. 2009 Mar;134(3):528-32 

[53] Pumera M (2008) Trends in analysis of explosives by microchip electrophoresis 

and conventional CE. Electrophoresis. 29(1):269-73. 

  

[54] Tagliaro F, Bortolotti F Recent advances in the applications of CE to forensic 

sciences (2005-2007). Electrophoresis [Electrophoresis], ISSN: 0173-0835, 

2008 Jan; Vol. 29 (1), pp. 260-8; 

  

[55] Pumera M.(2007) Trends in analysis of explosives by microchip electrophoresis 

and conventional CE. Electrophoresis. 2007 Nov 30 

  

[56] Hutchinson, Joseph P.; Evenhuis, Christopher J.; Johns, Cameron; Kazarian, 

Artaches A.; Breadmore, Michael C.; Macka, Miroslav; Hilder, Emily F.; Guijt, 

Rosanne M.; Dicinoski, Greg W.; Haddad, Paul R.. Identification of Inorganic 

Improvised Explosive Devices by Analysis of Postblast Residues Using 

Portable Capillary Electrophoresis Instrumentation and Indirect Photometric 

Detection with a Light-Emitting Diode. Analytical Chemistry, 9/15/2007, Vol. 79 

Issue 18, p7005-7013 

Gas Chromatography 

[57] Xiujuan Li, Zhaorui Zeng and Yi Zeng Solid-phase microextraction coupled to 

gas chromatography for the determination of 2,3-dimethyl-2,3-dinitrobutane as 

a marking agent for explosives Talanta 72.4 (June 15, 2007): p.1581(5). 

  

[58] Garrett Lee Burleson, Brittney Gonzalez, Kelsie Simons and Jorn C.C. Yu 

Forensic analysis of a single particle of partially burnt gunpowder by solid 

phase micro-extraction–gas chromatography-nitrogen phosphorus detector 

Journal of Chromatography A Volume 1216, Issue 22, 29 May 2009, Pages 

4679-4683 

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[59] Jeffrey A. Caulfield; Thomas J. Bruno; Keith E. Miller Enthalpy of Solution and 

Kováts Retention Indices for Nitroaromatic Compounds on Stationary Phases 

Using Gas Chromatography. Journal of Chemical & Engineering Data, 

Jun2009, Vol. 54 Issue 6, p1814-1822 

  

[60] Scott D. Harvey and Thomas J. Wenzel Selective retention of explosives and 

related compounds on gas-chromatographic capillary columns coated with 

lanthanide(III) β-diketonate polymers Journal of Chromatography A Volume 

1216, Issue 36, 4 September 2009, Pages 6417-6423 

Mass Spectrometry 

[61] Jitka Bečanová, Zdeněk Friedl and Zdeněk Šimek Identification and 

determination of trinitrotoluenes and their degradation products using liquid 

chromatography–electrospray ionization mass spectrometry International 

Journal of Mass Spectrometry Volume 291, Issue 3, 15 April 2010, Pages 133- 

139 

[62] E. Alizadeh, K. Graupner, A. Mauracher, S. Haughey, A. Edtbauer, M. Probst, 

T.D. Märk, T.A. Field and P. Scheier Electron attachment to 2-nitro-m-xylene 

International Journal of Mass Spectrometry Volume 289, Issues 2-3, 15 

January 2010, Pages 128-137 

[63] Schramm E, Hölzer J, Pütz M, Schulte-Ladbeck R, Schultze R, Sklorz M, Ulrich 

A, Wieser J, Zimmermann R. Real-time trace detection of security-relevant 

compounds in complex sample matrices by thermal desorption-single photon 

ionization-ion trap mass spectrometry (TD-SPI-ITMS) Anal Bioanal Chem. 

2009 Nov;395(6):1795-807 

[64] Scherperel, Gwynyth; Reid, Gavin E.; Smith, Ruth Waddell. Characterization 

of smokeless powders using nanoelectrospray ionization mass spectrometry 

(nESI-MS). Analytical & Bioanalytical Chemistry, Aug2009, Vol. 394 Issue 8, 

p2019-2028, 

  

[65] Huanwen Chen, Bin Hu, Yan Hu, Yanfu Huan, Zhiquan Zhou and Xiaolin Qiao 

Neutral Desorption Using a Sealed Enclosure to Sample Explosives on Human 

Skin for Rapid Detection by EESI-MS Journal of the American Society for 

Mass Spectrometry Volume 20, Issue 4, April 2009, Pages 719-722 

[66] Christopher Mullen, Michael J. Coggiola and Harald Oser Femtosecond Laser 

Photoionization Time-of-Flight Mass Spectrometry of Nitro-aromatic Explosives 

and Explosives Related Compounds Journal of the American Society for Mass 

Spectrometry Volume 20, Issue 3, March 2009, Pages 419-429 

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[67] Pablo Martínez-Lozano, Juan Rus, Gonzalo Fernández de la Mora, Marta 

Hernández and Juan Fernández de la Mora Secondary Electrospray Ionization 

(SESI) of Ambient Vapors for Explosive Detection at Concentrations Below 

Parts Per Trillion Journal of the American Society for Mass Spectrometry 

Volume 20, Issue 2, February 2009, Pages 287-294 

[68] Olivier L. Collin, Carolyn M. Zimmermann and Glen P. Jackson Fast gas 

chromatography negative chemical ionization tandem mass spectrometry of 

explosive compounds using dynamic collision-induced dissociation 


January 2009, Pages 93-99 

[69] Liguo Song, Andrew B. Dykstra, Huifang Yao and John E. Bartmess Ionization 

Mechanism of Negative Ion-Direct Analysis in Real Time: A Comparative Study 

with Negative Ion-Atmospheric Pressure Photoionization Journal of the 

American Society for Mass Spectrometry Volume 20, Issue 1, January 2009, 

Pages 42-50 

[70] Mauracher, P. Sulzer, E. Alizadeh, S. Denifl, F. Ferreira da Silva, M. Probst, 

T.D. Märk, P. Limão-Vieira and P. Scheier Electron attachment studies to 

musk ketone and high mass resolution anionic isobaric fragment detection 


November 2008, Pages 123-129 

[71] Talaty N, Mulligan CC, Justes DR, Jackson AU, Noll RJ, Cooks RG. Fabric 

analysis by ambient mass spectrometry for explosives and drugs. Analyst. 

2008 Nov;133(11):1532-40 

[72] Mitchell Wells, Michael J. Roth, Adam D. Keil, John W. Grossenbacher, Dina 

R. Justes, Garth E. Patterson and Dennis J. Barket Jr Implementation of 

DART and DESI Ionization on a Fieldable Mass Spectrometer Journal of the 

American Society for Mass Spectrometry Volume 19, Issue 10, October 2008, 

Pages 1419-1424 

[73] Aleem, A. Mauracher, P. Sulzer, S. Denifl, F. Zappa, A. Bacher, N. Wendt, T.D. 

Märk and P. Scheier (2008) Relative partial cross sections for anions formed 

upon electron attachment to nitrotoluene International Journal of Mass 

Spectrometry Volume 271, Issues 1-3, Pages 36-44 

[74] HUESTIS, DAVID L., MULLEN, CHRISTOPHER, COGGIOLA, MICHAEL J., 

OSER, HARALD LASER-IONIZATION MASS SPECTROMETRY OF 

EXPLOSIVES AND CHEMICAL WARFARE SIMULANTS. International Journal 

of High Speed Electronics & Systems; Mar2008, Vol. 18 Issue 1, p159-165 

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[75] E. Schramm; F. Mühlberger; S. Mitschke; G. Reichardt; R. Schulte-Ladbeck; M. 

Pütz; R. Zimmermann Determination of the Ionization Potentials of Security- 

Relevant Substances with Single Photon Ionization Mass Spectrometry Using 

Synchrotron Radiation. Applied Spectroscopy, Feb2008, Vol. 62 Issue 2, p238- 

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[76] Na N, Zhang C, Zhao M, Zhang S, Yang C, Fang X, Zhang X Direct detection 

of explosives on solid surfaces by mass spectrometry with an ambient ion 

source based on dielectric barrier discharge. Journal Of Mass Spectrometry: 

JMS [J Mass Spectrom], ISSN: 1076-5174, 2007 Aug; Vol. 42 (8), pp. 1079-85 

  

[77] Justes DR, Talaty N, Cotte-Rodriguez I, Cooks RG, Detection of explosives on 

skin using ambient ionization mass spectrometry. Chemical Communications 

(Cambridge, England) [Chem Commun (Camb)], ISSN: 1359-7345, 2007 Jun 

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[78] Williamson LN, Bartlett MG Quantitative liquid chromatography/time-of-flight 

mass spectrometry. Biomedical Chromatography: BMC [Biomed Chromatogr], 

ISSN: 0269-3879, 2007 Jun; Vol. 21 (6), pp. 567-76 

  

[79] S. Jönsson, L. Gustavsson and B. van Bavel (2007) Analysis of nitroaromatic 

compounds in complex samples using solid-phase microextraction and isotope 

dilution quantification gas chromatography–electron-capture negative ionisation 

mass spectrometry Journal of Chromatography A Volume 1164, Issues 1-2, 

Pages 65-73 

[80] Ochsenbein U, Zeh M, Berset JD. Comparing solid phase extraction and direct 

injection for the analysis of ultra-trace levels of relevant explosives in lake 

water and tributaries using liquid chromatography-electrospray tandem mass 

spectrometry. Chemosphere. 2008 Jun;72(6):974-80 

[81] Caroline McEnnis, Yamac Dikmelik and James B. Spicer (2007) Femtosecond 

laser-induced fragmentation and cluster formation studies of solid phase 

trinitrotoluene using time-of-flight mass spectrometry Applied Surface Science 

Volume 254, Issue 2, Pages 557-562 

[82] Philipp Sulzer, Andreas Mauracher, Stephan Denifl, Michael Probst, Tilmann D. 

Märk, Paul Scheier and Eugen Illenberger (2007) Probing di-nitrobenzene by 

low energy electrons Identification of isomers via resonances in dissociative 

electron attachment International Journal of Mass Spectrometry Volume 266, 

Issues 1-3, Pages 138-148 

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IRMS, Isotope Ratio MS 

[83] Anthony T. Quirk, John M. Bellerby, James F. Carter , Fay A. Thomas and 

Jenny C. Hill An initial evaluation of stable isotopic characterisation of postblast 

plastic debris from improvised explosive devices Science &Justice 

Volume 49, Issue 2, June 2009, Pages 87-93 

[84] Claire M. Lock and Wolfram Meier-Augenstein Investigation of isotopic linkage 

between precursor and product in the synthesis of a high explosive Forensic 

Science International Volume 179, Issues 2-3, 6 August 2008, Pages 157-162 

[85] David Widory, Jean-Jacques Minet and Martine Barbe-Leborgne Sourcing 

explosives: A multi-isotope approach Science &Justice Volume 49, Issue 2, 

June 2009, Pages 62-72 

[86] Anat Bernstein, Eilon Adar, Zeev Ronen, Harald Lowag, Willibald Stichler and 

Rainer U. Meckenstock Quantifying RDX biodegradation in groundwater using 

δ 15 N isotope analysis Journal of Contaminant Hydrology Volume 111, Issues 

1-4, 15 January 2010, Pages 25-35 

[87] Amaral HI, Fernandes J, Berg M, Schwarzenbach RP, Kipfer R. Assessing 

TNT and DNT groundwater contamination by compound-specific isotope 

analysis and 3H-3He groundwater dating: a case study in Portugal 

Chemosphere. 2009 Oct;77(6):805-12 

[88] Bernstein A, Ronen Z, Adar E, Nativ R, Lowag H, Stichler W, Meckenstock RU. 

Compound-specific isotope analysis of RDX and stable isotope fractionation 

during aerobic and anaerobic biodegradation. Environ Sci Technol. 2008 Nov 

1;42(21):7772-7. 

[89] V. F. Anisichkin Isotope studies of detonation mechanisms of TNT, RDX, and 

HMX Combustion, Explosion, and Shock Waves 43.5 (Sept 2007): p.580(7). 

  

[90] Anisichkin, V. F Isotope studies of detonation mechanisms of TNT, RDX, and 

HMX. Combustion, Explosion, & Shock Waves, Sep2007, Vol. 43 Issue 5, 

p580-586 

  

[91] N. Gentile, R.T.W. Siegwolf and O. Delémont Contribution of isotope ratio 

mass spectrometry to the investigation of improvised explosives: Isotopic study 

of black powders and ammonium nitrates Science &Justice Volume 50, Issue 

1, March 2010, Page 42 

[92] C. Lock and M. Beardah. Establishing Links Between Explosives Precursors 

and Products Using Isotope Ratio Mass Spectrometry. Science &Justice 

Volume 50, Issue 1, March 2010, Page 42 

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[93] Sarah J. Benson, Christopher J. Lennard, Philip Maynard, David M. Hill, Anita 

S. Andrew and Claude Roux. Forensic Analysis of Explosives Using Isotope 

Ratio Mass Spectrometry (IRMS) — Discrimination of Ammonium Nitrate 

Sources. Science &Justice Volume 49, Issue 2, June 2009, Pages 73-80 

  

[94] Sarah J. Benson, Christopher J. Lennard, Philip Maynard, David M. Hill, Anita 

S. Andrew and Claude Roux. Forensic Analysis of Explosives Using Isotope 

Ratio Mass Spectrometry (IRMS) — Preliminary Study on TATP and PETN 

Science &Justice Volume 49, Issue 2, June 2009, Pages 81-86 

[95] James F. Carter, Jenny C. Hill, Sean Doyle and Claire Lock Results of four 

inter-laboratory comparisons provided by the Forensic Isotope Ratio Mass 

Spectrometry (FIRMS) network Science &Justice Volume 49, Issue 2, June 

2009, Pages 127-137 

[96] Dianne Wakelin, Sean Doyle, Chris Andrews, Sarah Mountford and Niamh Nic 

Daeid (2008). Network developing forensic applications of stable Isotope Ratio 

Mass Spectrometry conference 2005 Science &Justice Volume 48, Issue 2, 

Pages 79-90 

General Spectroscopy: Fluorescence, Luminescence, Spectrophotometric, UV, 

Chemiluminescence 

[97] Qianqian Wang, Peter Jander, Cord Fricke-Begemann and Reinhard Noll. 

Comparison of 1064 nm and 266 nm excitation of laser-induced plasmas for 

several types of plastics and one explosive Spectrochimica Acta Part B: 

Atomic Spectroscopy Volume 63, Issue 10, October 2008, Pages 1011-1015 

[98] Christopher Ramos and Paul J. Dagdigian.Effect of photochemistry on 

molecular detection by cavity ringdown spectroscopy: case study of an 

explosive-related compound. Applied Optics 46.26 (Sept 10, 2007): p.6526(7). 

  

[99] Üzer, E. Erçağ and R. Apak (2008) Spectrophotometric determination of 

cyclotrimethylenetrinitramine (RDX) in explosive mixtures and residues with the 

Berthelot reaction Analytica Chimica Acta Volume 612, Issue 1, Pages 53-64 

[100] V.I. Babushok, F.C. Delucia, P.J. Dagdigian, J.L. Gottfried, C.A. Munson, M.J. 

Nusca and A.W. Miziolek. Kinetic modeling study of the laser-induced plasma 

plume of cyclotrimethylenetrinitramine (RDX) Spectrochimica Acta Part B: 

Atomic Spectroscopy 62.12 (Dec 2007): p.1321(8). 

  

[101] Pacheco-Londoño, Leonardo C.; Ortiz-Rivera, William; Primera-Pedrozo, Oliva 

M.; Hernández-Rivera, Samuel P. Vibrational spectroscopy standoff detection 

of explosives. Analytical & Bioanalytical Chemistry, Sep2009, Vol. 395 Issue 2, 

p323-335 

  

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[102] Erol Erçağ, Ayşem Üzer and Reşat Apak Selective spectrophotometric 

determination of TNT using a dicyclohexylamine-based colorimetric sensor 

Talanta Volume 78, Issue 3, 15 May 2009, Pages 772-780 

[103] Changmin Deng, Qingguo He, Jiangong Cheng, Defeng Zhu, Chao He and 

Tong Lin Unusual spectroscopic properties of PPE/TiO2 composite and its 

sensor response to TNT Synthetic Metals Volume 159, Issues 3-4, February 

2009, Pages 320-324 

[104] A.I. Nadezhdinskii, Ya.Ya. Ponurovskii and D.B. Stavrovskii. Non-contact 

detection of explosives by means of a tunable diode laser spectroscopy 

Applied Physics B: Lasers and Optics 90.2 (Feb 2008): p.361(4).   

  

[105] K. Chaudhary, A. M. Rudra, P. Kumbhakar and G. C. Bhar. Generation of 

coherent tunable deep UV radiation for detection and absorption studies of 

explosives RDX and TNT. Journal of Applied Spectroscopy 74.4 (July 

2007): p.571(7). 

  

[106] Sanchez JC, Toal SJ, Wang Z, Dugan RE, Trogler WC (2007) Selective 

Detection of Trace Nitroaromatic, Nitramine, and Nitrate Ester Explosive 

Residues Using a Three-Step Fluorimetric Sensing Process: A Tandem Turnoff, 

Turn-on Sensor. J Forensic Sci Nov2007, Vol. 52 Issue 6, p1308-1313 

  

[107] Shi GH, Shang ZB, Wang Y, Jin WJ, Zhang TC.(2007) Fluorescence 

quenching of CdSe quantum dots by nitroaromatic explosives and their relative 

compounds Spectrochim Acta A Mol Biomol Spectrosc. 2007 Aug 8 

  

[108] Germain ME, Knapp MJ. Optical explosives detection: from color changes to 

fluorescence turn-on. Chem Soc Rev. 2009 Sep;38(9):2543-55 

[109] Ramesh C. Sharma, Tracy S. Miller, Alexander D. Usachev, Jagdish P. Singh, 

Fang-Yu Yueh and David L. Monts Photo-fragmentation cross-section of 

gaseous 2,4,6-trinitrotoluene at different ultraviolet wavelengths 

Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy Volume 

72, Issue 3, April 2009, Pages 470-473 

[110] Zyryanov GV, Palacios MA, Anzenbacher P Jr. Simple molecule-based 

fluorescent sensors for vapor detection of TNT. Org Lett. 2008 Sep 

4;10(17):3681-4 

[111] Tammene Naddo a , Xiaomei Yang, Jeffrey S. Moore and Ling Zang Highly 

responsive fluorescent sensing of explosives taggant with an organic nanofibril 

film Sensors and Actuators B: Chemical Volume 134, Issue 1, 28 August 2008, 

Pages 287-291 

[112] Meaney MS, McGuffin VL Luminescence-based methods for sensing and 

detection of explosives. Anal Bioanal Chem. 2008 Aug;391(7):2557-76 

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[113] Monterola MP, Smith BW, Omenetto N, Winefordner JD. Photofragmentation 

of nitro-based explosives with chemiluminescence detection Anal Bioanal 

Chem. 2008 Aug;391(7):2617-26 

[114] Sheaff CN, Eastwood D, Wai CM, Addleman RS. Fluorescence detection and 

identification of tagging agents and impurities found in explosives. Appl 

Spectrosc. 2008 Jul;62(7):739-46 

[115] Adam R. Krause, Charles Van Neste, Larry Senesac, Thomas Thundat, and 

Eric Finot (2008) Trace explosive detection using photothermal deflection 

spectroscopy Appl. Phys. / Volume 103 / Issue 9 J. Appl. Phys. 103, 094906 

[116] Meaney, Melissa S., McGuffin, Victoria L. Investigation of common 

fluorophores for the detection of nitrated explosives by fluorescence quenching. 

Analytica Chimica Acta; Mar2008, Vol. 610 Issue 1, p57-67 

[117] Hughes AD, Glenn IC, Patrick AD, Ellington A, Anslyn EV. A pattern 

recognition based fluorescence quenching assay for the detection and 

identification of nitrated explosive analytes. Chemistry (Weinheim An Der 

Bergstrasse, Germany) [Chemistry], ISSN: 0947-6539, 2008; Vol. 14 (6), pp. 

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[118] Sushobhan Ghosh; Partha Sarathi Mukherjee Self-Assembly of a Nanoscopic 

Prism via a New Organometallic Pt3Acceptor and Its Fluorescent Detection of 

Nitroaromatics. Organometallics, Jan2008, Vol. 27 Issue 3, p316-319 

  

[119] Jason C. Sanchez; Antonio G. DiPasquale; Arnold L. Rheingold; William C. 

Trogler Synthesis, Luminescence Properties, and Explosives Sensing with 1,1- 

Tetraphenylsilole- and 1,1-Silafluorene-vinylene Polymers. Chemistry of 

Materials, Nov2007, Vol. 19 Issue 26, p6459-6470, 

[120] Andrew, Trisha L.; Swager, Timothy M A Fluorescence Turn-On Mechanism to 

Detect High Explosives RDX and PETN. Journal of the American Chemical 

Society, 6/13/2007, Vol. 129 Issue 23, p7254-7255, 

  

[121] Tammene Naddo, Yanke Che, Wei Zhang, Kaushik Balakrishnan, Xiaomei 

Yang, Max Yen, Jincai Zhao, Jeffrey S. Moore and Ling Zang Detection of 

explosives with a fluorescent nanofibril film Journal of the American Chemical 

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[122] Banas, K. Banas, M. Bahou, H.O. Moser, L. Wen, P. Yang, Z.J. Li, M. 

Cholewa, S.K. Lim and Ch.H. Lim Post-blast detection of traces of explosives 

by means of Fourier transform infrared spectroscopy Vibrational Spectroscopy 

Volume 51, Issue 2, 10 November 2009, Pages 168-176 

[123] S.P. Sharmaand S.C. Lahiri (2008) Absorption spectroscopic and FTIR studies 

on EDA complexes between TNT (2,4,6-trinitrotoluene) with amines in DMSO 

and determination of the vertical electron affinity of TNT Spectrochimica Acta 

Part A: Molecular and Biomolecular Spectroscopy Volume 70, Issue 1, Pages 

144-153 

[124] Mou, Yongyan; Rabalais, J. Wayne Detection and Identification of Explosive 

Particles in Fingerprints Using Attenuated Total Reflection-Fourier Transform 

Infrared Spectromicroscopy. Journal of Forensic Sciences Jul2009, Vol. 54 

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Raman Spectroscopy 

[125] Moros J, Lorenzo JA, Lucena P, Tobaria LM, Laserna JJ. Simultaneous 

Raman spectroscopy-laser-induced breakdown spectroscopy for instant 

standoff analysis of explosives using a mobile integrated sensor platform Anal 

Chem. 2010 Feb 15;82(4):1389-400 

[126] Alison J. Hobro and Bernhard Lendl Stand-off Raman spectroscopy TrAC 

Trends in Analytical Chemistry Volume 28, Issue 11, December 2009, Pages 

1235-1242 

[127] Emmons ED, Tripathi A, Guicheteau JA, Christesen SD, Fountain AW 3rd. 

Raman chemical imaging of explosive-contaminated fingerprints Appl 

Spectrosc. 2009 Nov;63(11):1197-203 

[128] Y. Fleger, L. Nagli, M. Gaft and M. Rosenbluh Narrow gated Raman and 

luminescence of explosives Journal of Luminescence Volume 129, Issue 9, 

September 2009, Pages 979-983 

[129] Esam M.A. Ali, Howell G.M. Edwards and Ian J. Scowen In-situ detection of 

single particles of explosive on clothing with confocal Raman microscopy 

Talanta Volume 78, Issue 3, 15 May 2009, Pages 1201-1203 

[130] Comanescu G, Manka CK, Grun J, Nikitin S, Zabetakis D. Identification of 

explosives with two-dimensional ultraviolet resonance Raman spectroscopy. 

Appl Spectrosc. 2008 Aug;62(8):833-9 

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[131] Alexander Portnov, Salman Rosenwaks, and Ilana Bar Detection of particles of 

explosives via backward coherent anti-Stokes Raman spectroscopy Appl. 

Phys. Lett. / Volume 93 / Issue 4 041115 31 July 2008 

[132] Gaft and L. Nagli UV gated Raman spectroscopy for standoff detection of 

explosives Optical Materials Volume 30, Issue 11, July 2008, Pages 1739- 

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[133] L. Nagli, M. Gaft, Y. Fleger and M. Rosenbluh Absolute Raman cross-sections 

of some explosives: Trend to UV Optical Materials Volume 30, Issue 11, July 

2008, Pages 1747-1754 

[134] Katz, A. Natan Y. Silberberg, and S. Rosenwaks (2008) Standoff detection of 

trace amounts of solids by nonlinear Raman spectroscopy using shaped 

femtosecond pulses Appl. Phys. Lett. / Volume 92 / Issue 17 171116 (April 28) 

[135] Li H, Harris DA, Xu B, Wrzesinski PJ, Lozovoy VV, Dantus M Coherent modeselective 

Raman excitation towards standoff detection. Opt Express. 2008 Apr 

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[136] C. Eliasson, N.A. Macleod and P. Matousek Noninvasive detection of 

concealed liquid explosives using Raman spectroscopy. Analytical 

Chemistry 79.21 (Nov 1, 2007): p.8185(5). 

  

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[137] Korošec, Romana; Kajič, Petra; Bukovec, P. Determination of water, 

ammonium nitrate and sodium nitrate content in ‘water-in-oil’ emulsions using 

TG and DSC. Journal of Thermal Analysis & Calorimetry, Aug2007, Vol. 89 

Issue 2, p619-624 

  

Medical; Pathology; Injuries; Toxicology 

[138] Szakal, Christopher; Brewer, Tim M Analysis and Mechanisms of 

Cyclotrimethylenetrinitramine Ion Formation in Desorption Electrospray 

Ionization. Analytical Chemistry, 7/1/2009, Vol. 81 Issue 13, p5257-5266 

  

[139] Baohong Zhang and Xiaoping Pan RDX Induces Aberrant Expression of 

MicroRNAs in Mouse Brain and Liver Environ Health Perspect. 2009 February; 

117(2): 231–240. (This article is available online) 

[140] K.R. Nagesh MD, Associate Professor, Ritesh G. Menezes MD DNB, Associate 

Professor, B. Suresh Kumar Shetty MD, Associate Professor and Anand 

Menon MD, Professor and Head Circumscribed injuries caused by an 

explosion in firecracker factory Journal of Forensic and Legal Medicine 

Volume 17, Issue 3, April 2010, Pages 169-171 

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[141] Mark Lenz, Ralf Schmidt, Thomas Muckley, Torsten Donicke, Reinhard Friedel 

and Gunther O Hofmann Wheelbarrow tire explosion causing trauma to the 

forearm and hand: a case report Journal of Medical Case Reports 2009, 3:129 

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[142] Stephen J Wolf MD, Maj Vikhyat S Bebarta MD, USAF, Carl J Bonnett MD, 

Prof Peter T Pons MD and Stephen V Cantrill Blast injuries The Lancet 

Volume 374, Issue 9687, 1 August 2009-7 August 2009, Pages 405-415 

[143] Deborah L. Warden, Louis M. French, Leslie Shupenko, Jamie Fargus, Gerard 

Riedy, Marleigh E. Erickson, Michael S. Jaffee and David F. Moore Case 

report of a soldier with primary blast brain injury NeuroImage Volume 47, 

Supplement 2, August 2009, Pages T152-T153 

[144] Cockerham, Glenn C.; Goodrich, Gregory L.; Weichel, Eric D.; Orcutt, James 

C.; Rizzo, Joseph F.; Bower, Kraig S.; Schuchard, Ronald A Eye and visual 

function in traumatic brain injury. Journal of Rehabilitation Research & 

Development, 2009, Vol. 46 Issue 6, p811-818 

  

[145] V. Macko, L. Straka and J. Krajcovic C-13 explosion victims – peculiarities of 

autopsy findings Legal Medicine Volume 11, Supplement 1, April 2009, Pages 

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[146] Yoko Ihama, Tetsuji Miyazaki, Chiaki Fuke and Zenshin Taira A case of death 

from the explosion of a 66 mm M72 High Explosive Anti-Tank rocket Legal 

Medicine Volume 10, Issue 4, July 2008, Pages 210-215 

[147] Johnson MS, Quinn MJ, Bazar MA, Gust KA, Escalon BL, Perkins EJ (2007) 

Subacute toxicity of oral 2,6-dinitrotoluene and 1,3,5-trinitro-1,3,5-triazine 

(RDX) exposure to the northern bobwhite (Colinus virginianus). Environ Toxicol 

Chem. 26(7):1481-7.  

  

[148] Fausti, Stephen A.; Wilmington, Debra J.; Gallun, Frederick J.; Myers, Paula J.; 

Henry, James A Auditory and vestibular dysfunction associated with blastrelated 

traumatic brain injury. Journal of Rehabilitation Research & 

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[149] Weil YA, Peleg K, Givon A; Israeli Trauma Group, Mosheiff R. Musculoskeletal 

injuries in terrorist attacks--a comparison between the injuries sustained and 

those related to motor vehicle accidents, based on a national registry database 

Injury. 2008 Dec;39(12):1359-64 

[150] Ramasamy A, Harrisson SE, Clasper JC, Stewart MP. Injuries from roadside 

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[151] Ulrich M. Rieger, Daniel F. Kalbermatten, Reto Wettstein, Ilonka Heider, Martin 

Haug and Gerhard Pierer (2008) Marjolin's ulcer revisited – basal cell 

carcinoma arising from grenade fragments? Case report and review of the 

literature Journal of Plastic, Reconstructive & Aesthetic Surgery Volume 61, 

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[152] D. Potocnjak, R. Bari&cacute;-Rafaj, N. Lemo, V. Matijatko, I. Kis, V. 

Mrljak and I. Harapin. Poisoning of a dog with the explosive pentaerythrityl 

tetranitrate Journal of Small Animal Practice 49.6 (June 2008): p.314(5). 

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[153] S.D. Gosling and S.J. Hilliard Personality and performance in explosivedetection 

military working dogs (MWDs) Journal of Veterinary Behavior: 

Clinical Applications and Research Volume 4, Issue 6, November-December 

2009, Pages 239-240 

[154] L.J. Myers, E. Mueller and W. Weimar Reliability of operational detector dog 

handler teams and associated factors Journal of Veterinary Behavior: Clinical 

Applications and Research Volume 4, Issue 6, November-December 2009, 

Page 252 (No abstract available) 

[155] W.A. Kafka SOKKS ® micro amount animal odor conditioning Journal of 

Veterinary Behavior: Clinical Applications and Research Volume 4, Issue 6, 

November-December 2009, Pages 244-245 (No abstract available) 

[156] William S. Helton (2007) Skill in Expert Dogs Journal of Experimental 

Psychology: Applied Volume 13, Issue 3 Pages 171-178 

  

[157] Aspects of Explosives Detection Chapter 3 - Detection of Explosives by Dogs 

J.C. Oxley and L.P. Waggoner Available online 18 February 2009. Copyright © 

2009 Elsevier B.V. All rights reserved. Edited by: Maurice Marshall and Jimmie 

C. Oxley.ISBN: 978-0-12-374533-0  

[158] Luis Alberto Marin Guerrero Quick Learning Techniques: Dogs used to detect 

narcotics and explosives Journal of Veterinary Behavior: Clinical Applications 

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Biosensors 

[159] Ehrentreich-Förster E, Orgel D, Krause-Griep A, Cech B, Erdmann VA, Bier F, 

Scheller FW, Rimmele M Biosensor-based on-site explosives detection using 

aptamers as recognition elements. Anal Bioanal Chem. 2008 Jul;391(5):1793- 

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[160] Smith RG; D'Souza N; Nicklin S A review of biosensors and biologicallyinspired 

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Detection; LIBS 

[161] R. González; P. Lucena; L. M. Tobaria; J. J. Laserna Standoff LIBS Detection 

of Explosive Residues Behind a Barrier JAAS (Journal of Analytical Atomic 

Spectrometry), Jul2009, Vol. 24 Issue 8, p1123-1126 

  

[162] V. Lazic, A. Palucci, S. Jovicevic, C. Poggi and E. Buono. Analysis of 

explosive and other organic residues by laser induced breakdown 

spectroscopy Spectrochimica Acta Part B: Atomic Spectroscopy Volume 64, 

Issue 10, October 2009, Pages 1028-1039 

[163] Wallin, Sara; Pettersson, Anna; Östmark, Henric; Hobro, Alison Laser-based 

standoff detection of explosives: a critical review. Analytical & Bioanalytical 


  

[164] Gottfried, Jennifer L.; De Lucia Jr., Frank C.; Munson, Chase A.; Miziolek, 

Andrzej W. Laser-induced breakdown spectroscopy for detection of explosives 

residues: a review of recent advances, challenges, and future prospects. 

Analytical & Bioanalytical Chemistry, Sep2009, Vol. 395 Issue 2, p283-300 

  

[165] Gottfried JL; De Lucia FC Jr; Munson CA; Miziolek AW Standoff detection of 

chemical and biological threats using laser-induced breakdown spectroscopy. 

Applied Spectroscopy [Appl Spectrosc] 2008 Apr; Vol. 62 (4), pp. 353-63 

[166] Jennifer L. Gottfried; Frank C. De Lucia Jr; Chase A. Munson; Andrzej W. 

Miziolek Strategies for residue explosives detection using laser-induced 

breakdown spectroscopy. JAAS (Journal of Analytical Atomic Spectrometry), 

Jan2008, Vol. 23 Issue 2, p205-216, 

  

[167] Frank C. De Lucia, Jennifer L. Gottfried, Chase A. Munson and Andrzej W. 

Miziolek Double pulse laser-induced breakdown spectroscopy of explosives: 

Initial study towards improved discrimination Spectrochimica Acta Part B: 

Atomic Spectroscopy 62.12 (Dec 2007): p.1399(6). 

  

[168] Jennifer L. Gottfried, Frank C. De Lucia, Chase A. Munson and Andrzej W. 

Miziolek. Double-pulse standoff laser-induced breakdown spectroscopy for 

versatile hazardous materials detection. Spectrochimica Acta Part B: Atomic 

Spectroscopy 62.12 (Dec 2007): p.1405(7). 

  

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[169] Christian Bohling, Konrad Hohmann, Dirk Scheel, Christoph Bauer, Wolfgang 

Schippers, Jorg Burgmeier, Ulrike Willer, Gerhard Holl, Wolfgang Schade Allfiber-coupled 

laser-induced breakdown spectroscopy sensor for hazardous 

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Spectroscopy 62.12 (Dec 2007): p.1519(9). 

  

[170] Moros J, Lorenzo JA, Lucena P, Tobaria LM, Laserna JJ. Simultaneous 

Raman spectroscopy-laser-induced breakdown spectroscopy for instant 

standoff analysis of explosives using a mobile integrated sensor platform Anal 

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Detection: Neutron 

[171] Vladivoj Valkovic, Davorin Sudac and Dario Matika Fast neutron sensor for 

detection of explosives and chemical warfare agents Applied Radiation and 

Isotopes Volume 68, Issues 4-5, April-May 2010, Pages 888-892 

[172] V A Gribkov, S V Latyshev, R A Miklaszewski, M Chernyshova, K Drozdowicz, 

U Wiącek, K Tomaszewski and B D Lemeshko. A dense plasma focus-based 

neutron source for a single-shot detection of illicit materials and explosives by a 

nanosecond neutron pulse Physica Scripta Volume 81, Number 3 035502 

[173] Hee-Jung Im; Kyuseok Song. Applications of Prompt Gamma Ray Neutron 

Activation Analysis: Detection of Illicit Materials. Applied Spectroscopy 

Reviews, Jul/Aug2009, Vol. 44 Issue 4, p317-334 

  

[174] D. Koltick and E. Sword A Neutron Based Scanner to Detect Explosives in 

Small, Sealed Containers AIP Conf. Proc. -- March 10, 2009 -- Volume 1099, 

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[175] E. H. Seabury, D. L. Chichester, C. J. Wharton, and A. J. Caffrey A 

Comparison of Neutron-Based Non-Destructive Assessment Methods for 

Chemical Warfare Materiel and High Explosives AIP Conf. Proc. -- March 10, 

2009 -- Volume 1099, pp. 928-931 

[176] D Vartsky, G Feldman, I Mor, M B Goldberg, D Bar and V Dangendorf Signal 

and noise analysis in TRION—Time-Resolved Integrative Optical Fast Neutron 

detector JINST 4 P02001 16 February 2009 (open access article) 

[177] M D Aspinall, M J Joyce, R O Mackin, Z Jarrah, A J Boston, P J Nolan, A J 

Peyton and N P Hawkes Sample-interpolation timing: an optimized technique 

for the digital measurement of time of flight for γ rays and neutrons at relatively 

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[178] Viesti, G.; Botosso, C.; Fabris, D.; Lunardon, M.; Moretto, S.; Nebbia, G.; 

Pesente, S.; Zenoni, A.; Donzella, A.; Perot, B.; Carasco, C.; Bernard, S.; 

Mariani, A.; Szabo, J.-L.; Sannie, G.; Valkovic, V.; Sudac, D.; Nad, K.; Peerani, 

P.; Sequeira, V.. Scanning Cargo Containers with Tagged Neutrons. AIP 

Conference Proceedings, 10/26/2007, Vol. 947 Issue 1, p57-62, 

  

[179] Forero, N. C.; Cruz, A. H.; Cristancho, F Simulation of Neutron Backscattering 

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[180] D. Rezaei Ochbelagh, H. Miri Hakimabad and R. Izadi Najafabadi The soil 

moisture and its effect on the detection of buried hydrogenous material by 

neutron backscattering technique Radiation Physics and Chemistry 

Volume 78, Issue 5, May 2009, Pages 303-306 

[181] Maxim Karetnikov, Anatoly Klimov, Sergey Korotkov, Evgeny Meleshko, Igor 

Ostashev, Timur Khasaev and Guenrikh Yakovlev (2007) Efficiency of 

background suppression by tagged neutron technology Nuclear Instruments 

and Methods in Physics Research Section B: Beam Interactions with Materials 

and Atoms Volume 261, Issues 1-2 Pages 307-310 

Detection: Terahertz 

[182] Appleby, Roger; Wallace, H. Bruce Standoff Detection of Weapons and 

Contraband in the 100 GHz to 1 THz Region. IEEE Transactions on Antennas 

& Propagation, Nov2007 Part 1 of 2, Vol. 55 Issue 11, p2944-2956 

  

[183] Hua Zhong; Redo-Sanchez, Albert; Xi-Cheng Zhang. STANDOFF SENSING 

AND IMAGING OF EXPLOSIVE RELATED CHEMICAL AND BIO-CHEMICAL 

MATERIALS USING THz-TDS. International Journal of High Speed Electronics 

& Systems, Jun2007, Vol. 17 Issue 2, p239-249, 

  

[184] YUNQING CHEN; HAIBO LIU; ZHANG, X. –C EXPERIMENTAL AND 

DENSITY FUNCTIONAL THEORY STUDY ON THz SPECTRA OF 4-NT AND 

2, 6-DNT. International Journal of High Speed Electronics & Systems, 

Jun2007, Vol. 17 Issue 2, p283-291 

  

[185] Michael J. Fitch, Megan R. Leahy-Hoppa, Edward W. Ott and Robert Osiander 

(2007) Molecular absorption cross-section and absolute absorptivity in the THz 

frequency range for the explosives TNT, RDX, HMX, and PETN Chemical 

Physics Letters Volume 443, Issues 4-6, Pages 284-288 

  

[186] Leahy-Hoppa, Megan R.; Fitch, Michael J.; Osiander, Robert Terahertz 

spectroscopy techniques for explosives detection. Analytical & Bioanalytical 


  

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[187] Zhou Zhen, Chen An-Tao and Feng Li-Shuang Studies on 2,4-DNT Mixtures 

Using Reflection Terahertz Time Domain Spectroscopy for Explosives 

Detection Chinese Phys. Lett. 26 037801 Issue 3 (March 2009) 

[188] Damian G. Allis, J. Axel Zeitler, Philip F. Taday and Timothy M. Korter 

Theoretical analysis of the solid-state terahertz spectrum of the high explosive 

RDX Chemical Physics Letters Volume 463, Issues 1-3, 22 September 2008, 

Pages 84-89 

[189] Joseph S. Melinger, N. Laman, and D. Grischkowsky The underlying terahertz 

vibrational spectrum of explosives solids Appl. Phys. Lett. / Volume 93 / Issue 

1 011102 8 July 2008 

[190] Liangliang Zhang, Hua Zhong, Chao Deng, Cunlin Zhang, and Yuejin Zhao 

(2008) Terahertz wave reference-free phase imaging for identification of 

explosives Appl. Phys. Lett. / Volume 92 / Issue 9 / 091117 

[191] Giles Davies, Andrew D. Burnett, Wenhui Fan, Edmund H. Linfield and John E. 

Cunningham (2008) Terahertz spectroscopy of explosives and drugs Materials 

Today Volume 11, Issue 3, Pages 18-26 

[192] Alexander Sinyukov, Ivan Zorych, Zoi-Heleni Michalopoulou, Dale Gary, Robert 

Baratand John F. Federici (2008). Detection of explosives by Terahertz 

synthetic aperture imaging—focusing and spectral classification (Détection 

d'explosives par l'imagerie térahertz à ouverture synthétique : mise au point et 

classification spectrale) Comptes Rendus Physique Volume 9, Issue 2, Pages 

248-261 

[193] Yingxin Wang, Ziran Zhao, Zhiqiang Chen, Kejun Kang, Bing Feng and Yan 

Zhang. Terahertz absorbance spectrum fitting method for quantitative detection 

of concealed contraband Journal of Applied Physics 102.11 (Dec 1, 

2007): p.113108-1-113108-6 

  

[194] Zimdars, david; white, jeffrey; stuk, g.; sucha, g.; fichter, g.; williamson, s. L 

time domain terahertz imaging of threats in luggage and personnel. 

International Journal of High Speed Electronics & Systems, Jun2007, Vol. 17 

Issue 2, p271-281 

  

[195] Kemp, michael c.; glauser, antony; baker, colin multi-spectral terahertz imaging 

using reflected and scattered radiation. International Journal of High Speed 

Electronics & Systems, Jun2007, Vol. 17 Issue 2, p403-414, 

  

[196] J. Hooper, E. Mitchell, C. Konek and J. Wilkinson Terahertz optical properties 

of the high explosive β-HMX Chemical Physics Letters Volume 467, Issues 4- 

6, 5 January 2009, Pages 309-312 

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[197] Ortolani, J. S. Lee, U. Schade, and H.-W. Hübers Surface roughness effects 

on the terahertz reflectance of pure explosive materials Appl. Phys. Lett. / 

Volume 93 / Issue 8 081906 25 August 2008 

[199] Qing-li Zhou, Cun-lin Zhang, Kai-jun Mu, Bin Jin, Liang-liang Zhang, Wei-wei 

Li, and Rui-shu Feng (2008) Optical property and spectroscopy studies on the 

explosive 2,4,6-trinitro-1,3,5-trihydroxybenzene in the terahertz range Appl. 

Phys. Lett. / Volume 92 / Issue 10 101106 (March 10) 

Detection; Nuclear Techniques, NQR, Quadrupole Resonance 

[200] Hong Xiong, Jian Li and Geoffrey A. Barrall Joint TNT and RDX detection via 

quadrupole resonance. IEEE Transactions on Aerospace and Electronic 

Systems 43.4 (Oct 2007): p.1282(12). 

  

[201] Alan Gregorovič and Tomaž Apih TNT detection with 14 N NQR: Multipulse 

sequences and matched filter Journal of Magnetic Resonance Volume 198, 

Issue 2, June 2009, Pages 215-221 

[202] Jakobsson, Andreas; Mossberg, Magnus Using Spatial Diversity to Detect 

Narcotics and Explosives Using NQR Signals. IEEE Transactions on Signal 

Processing, Sep2007, Vol. 55 Issue 9, p4721-4726 

[203] S.D. Somasundaram, A. Jakobsson and J.A.S. Smith (2008) Analysis of 

nuclear quadrupole resonance signals from mixtures Signal Processing 


[204] Janko Luznik, Janez Pirnat, Vojko Jazbinsek, Tomaz Apih, Robert Blinc, Janez 

Seliger and Zvonko Trontelj. Improved [super 14]N nuclear quadrupole 

resonance detection of trinitrotoluene using polarization transfer from protons 

to [super 14]N nuclei. Journal of Applied Physics 102.8 (Oct 15, 

2007): p.084903-1-084903-7. 

  

[205] Daren R. Norman, James L. Jones, Brandon W. Blackburn, Kevin J. Haskell, 

James T. Johnson, Scott M. Watson, Alan W. Hunt, Randy Spaulding, Frank 

Harmon Time-dependent delayed signatures from energetic photon 

interrogations Nuclear Inst. and Methods in Physics Research, B 262.1 (August 


  

[206] William Bertozzi, Stephen E. Korbly, Robert J. Ledoux and William Park 

Nuclear resonance fluorescence and effective Z determination applied to 

detection and imaging of special nuclear material, explosives, toxic substances 

and contraband Nuclear Inst. and Methods in Physics Research, 

B 262.1 (August 2007): p.331(6). 

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[207] C.E. Demonchy, M. Caamaño, H. Wang, W. Mittig, P. Roussel-Chomaz, H. 

Savajols, M. Chartier, D. Cortina-Gil, A. Fomichev, G. Frémont, P. Gangnant, 

A. Gillibert, L. Giot, M.S. Golovkov, B. Jurado, J.F. Libin, A. Obertelli, E. 

Pollaco, A. Rodin, Ch. Spitaels, S. Stepantsov, G. Ter-Akopian and R. Wolski 

(2007) MAYA: An active-target detector for binary reactions with exotic beams 

Nuclear Instruments and Methods in Physics Research Section A: 

Accelerators, Spectrometers, Detectors and Associated Equipment Volume 

583, Issues 2-3 Pages 341-349 

[208] Davorin Sudac, Sasa Blagus and Vladivoj Valkovic (2007) The limitations of 

associated alpha particle technique for contraband container inspections 

Nuclear Instruments and Methods in Physics Research Section B: Beam 

Interactions with Materials and Atoms Volume 263, Issue 1 Pages 123-126 

[209] W.L. Dunn, C.J. Solomon, K.W. Loschke, D.A. Bradley and W.B. Gilboy (2007) 

Ionizing photon methods for standoff bomb detection Nuclear Instruments and 

Methods in Physics Research Section A: Accelerators, Spectrometers, 

Detectors and Associated Equipment Volume 580, Issue 1 Pages 778-781 

[210] K.A. Ten, V.M. Aulchenko, L.A. Lukjanchikov, E.R. Pruuel, L.I. Shekhtman, 

B.P. Tolochko, I.L. Zhogin and V.V. Zhulanov Application of introduced nanodiamonds 

for the study of carbon condensation during detonation of condensed 

explosives Nuclear Instruments and Methods in Physics Research Section A: 


603, Issues 1-2, 11 May 2009, Pages 102-104 

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Farrell, A. Khilchenko, V. Kobets, S. Konstantinov, A. Krivenko, A. 

Kudryavtsev, M. Tiunov, V. Savkin, V. Shirokov and I. Sorokin (2007) Status of 

BINP proton tandem accelerator Nuclear Instruments and Methods in Physics 

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[212] Daren R. Norman, James L. Jones, Brandon W. Blackburn, Kevin J. Haskell, 

James T. Johnson, Scott M. Watson, Alan W. Hunt, Randy Spaulding and 

Frank Harmon (2007) Time-dependent delayed signatures from energetic 

photon interrogations Nuclear Instruments and Methods in Physics Research 

Section B: Beam Interactions with Materials and Atoms Volume 261, Issues 1-2 

Pages 316-320 

[213] King, Michael Joseph; Miller, Gill T.; Reijonen, Jani; Qing Ji; Andresen, Nord; 

Gicquel, Frederic; Kalvas, Taneli; Ka-Ngo Leung; Kwan, Joe W Initial 

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Brian A. Collins Photon and neutron interrogation techniques for chemical 

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J.-L. Szabo, G. Sannie, M. Lunardon, C. Bottosso, S. Moretto, S. Pesente, P. 

Peerani, V. Sequeira and M. Salvato (2007) Photon attenuation and neutron 

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S. Pesente (2007) Fast neutron inspection of sea containers for the presence 

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B: Beam Interactions with Materials and Atoms Volume 263, Issue 1, Pages 

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A. Zenoni, G. Bonomi, A. Donzella, A. Fontana, G. Boghen, V. Valkovic, D. 

Sudac, M. Moszynski, T. Batsch, M. Gierlik, D. Woski, W. Klamra, P. Isaksson, 

P. Le Tourneur, M. Lhuissier, A. Colonna, C. Tintori, P. Peerani, V. Sequeira 

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inspection system Nuclear Instruments and Methods in Physics Research 

Section B: Beam Interactions with Materials and Atoms Volume 261, Issues 1- 

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(2007) Portable fast-neutron radiography with the nuclear materials 

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Atoms Volume 261, Issues 1-2, Pages 387-390 

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V. Valkovic, D. Sudac, G. Viesti, M. Lunardon, C. Bottosso, D. Fabris, G. 

Nebbia, S. Pesente, S. Moretto, A. Zenoni, A. Donzella, M. Moszynski, M. 

Gierlik, T. Batsch, D. Wolski, W. Klamra, P. Le Tourneur, M. Lhuissier, A. 

Colonna, C. Tintori, P. Peerani, V. Sequeira and M. Salvato (2008) In-field tests 

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(2007) Comparison of DD, DT and Cf-252 neutron excitation of light and 

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[224] Xiao-Hong Li, Rui-Zhou Zhang, Xiang-Dong Yang and Hong Zhang (2007) 

Density functional studies of the heats of formation of several nitroester 

compounds Journal of Molecular Structure: THEOCHEM 

[225] Alexandros Paraskevas and Beverley Arendell (2007) A strategic framework for 

terrorism prevention and mitigation in tourism destinations Tourism 

Management Volume 28, Issue 6 Pages 1560-1573 

[226] Anthony Dicken, Keith Rogers, Paul Evans, Joseph Rogers and Jer Wang 

Chan The separation of X-ray diffraction patterns for threat detection Applied 

Radiation and Isotopes Volume 68, Issue 3, March 2010, Pages 439-443 

[227] Y. Liu, B.D. Sowerbyand J.R. Tickner (2008) Comparison of neutron and highenergy 

X-ray dual-beam radiography for air cargo inspection Applied Radiation 

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X-ray dual-beam radiography for air cargo inspection. Applied 

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[229] Mahmoud Tabrizchi and Vahideh ILbeigi Detection of explosives by positive 

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Volume 176, Issues 1-3, 15 April 2010, Pages 692-696 

[230] Babis, J. S.; Sperline, R. P.; Knight, A. K.; Jones, D. A.; Gresham, C. A.; 

Denton, M. B Performance evaluation of a miniature ion mobility spectrometer 

drift cell for application in hand-held explosives detection ion mobility 

spectrometers. Analytical & Bioanalytical Chemistry, Sep2009, Vol. 395 Issue 

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[231] Joshi, Monica; Delgado, Yisenny; Guerra, Patricia; Lai, Hanh; Almirall, José R 

Detection of odor signatures of smokeless powders using solid phase 

microextraction coupled to an ion mobility spectrometer. Forensic Science 

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[232] Melanie J. Waltman, Prabha Dwivedi, Herbert H. Hill Jr, William C. Blanchard 

and Robert G. Ewing Characterization of a distributed plasma ionization 

source (DPIS) for ion mobility spectrometry and mass spectrometry Talanta 

Volume 77, Issue 1, 19 October 2008, Pages 249-255 

[233] Guerra P, Lai H, Almirall JR Analysis of the volatile chemical markers of 

explosives using novel solid phase microextraction coupled to ion mobility 

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[234] Oxley, Jimmie C. Smith, James L., Kirschenbaum, Louis J., Marimganti, 

Suvarna, Vadlamannati, Sravanthi Detection of Explosives in Hair Using Ion 

Mobility Spectrometry Journal of Forensic Sciences; May2008, Vol. 53 Issue 3, 

p690-693 

[235] Lai H, Guerra P, Joshi M, Almirall JR Analysis of volatile components of drugs 

and explosives by solid phase microextraction-ion mobility spectrometry. 

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[236] Michael Martin, Mark Crain, Kevin Walsh, R. Andrew McGill, Eric Houser, 

Jennifer Stepnowski, Stanley Stepnowski, Huey-Daw Wu, Stuart Ross 

Microfabricated vapor preconcentrator for portable ion mobility spectroscopy 

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[237] Stefan Zimmermann, Nora Abel, Wolfgang Baether and Sebastian Barth An 

ion-focusing aspiration condenser as an ion mobility spectrometer Sensors & 

Actuators: B. Chemical 125.2 (August 8, 2007): p.428(7). 

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[238] Abu B. Kanu and Herbert H. Hill Jr (2007) Identity confirmation of drugs and 

explosives in ion mobility spectrometry using a secondary drift gas Talanta 

Volume 73, Issue 4 Pages 692-699 

[239] Stefan Zimmermann, Nora Abel, Wolfgang Baether and Sebastian Barth (2007) 

An ion-focusing aspiration condenser as an ion mobility spectrometer Sensors 

and Actuators B: Chemical Volume 125, Issue 2, Pages 428-434 

[240] Kolakowski BM, Mester Z.(2007) Review of applications of high-field 

asymmetric waveform ion mobility spectrometry (FAIMS) and differential 

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[241] Lu, Sijun; Zhang, Jian; Dagan Feng, David Detecting unattended packages 

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R. Papantonakis, V. Nguyen, G. K. Hubler, and R. A. McGill Stand-off 

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Egger and Paul J. Petersen Idaho Explosives Detection System: Development 

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Trogler Lewis acid–base interactions enhance explosives sensing in silacycle 

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[245] Xinghua Xie , Jing Zhu and Huisheng Zhou Deflagration products from 

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[246] Willer, Ulrike; Schade, Wolfgang Photonic sensor devices for explosive 

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Li J, Tao N. A hybrid electrochemical-colorimetric sensing platform for 

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[248] A.T. Nimal, Upendra Mittal, Mohan Singh, Mamta Khaneja, G.K. Kannan, J.C. 

Kapoor, Vinita Dubey, P.K. Gutch, Gobardhan Lal, K.D. Vyas and D.C. Gupta. 

Development of handheld SAW vapor sensors for explosives and CW agents 

Sensors and Actuators B: Chemical Volume 135, Issue 2, 15 January 2009, 

Pages 399-410 

[249] Li Zheng and Jun-feng Song Curcumin multi-wall carbon nanotubes modified 

glassy carbon electrode and its electrocatalytic activity towards oxidation of 

hydrazine Sensors and Actuators B: Chemical Volume 135, Issue 2, 15 


[250] Ravishankar S. Dudhe, S. P. Tiwari, Harshil N. Raval, Mrunal A. Khaderbad, 

Rahul Singh, Jasmine Sinha, M. Yedukondalu, M. Ravikanth, Anil Kumar, and 

V. Ramgopal Rao Explosive vapor sensor using poly (3-hexylthiophene) and 

Cu II tetraphenylporphyrin composite based organic field effect transistors Appl. 

Phys. Lett. / Volume 93 / Issue 26 / (Dec 31) 

[251] Irena Grigoriants, Boris Markovsky, Rachel Persky, Ilana Perelshtein, Aharon 

Gedanken, Doron Aurbach, Boris Filanovsky, Tatiana Bourenko and Israel 

Felner Electrochemical reduction of trinitrotoluene on core–shell tin–carbon 

electrodes Electrochimica Acta Volume 54, Issue 2, 30 December 2008, 

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[252] Dechang Yi, Anders Greve, Jan H. Hales, Larry R. Senesac, Zachary J. Davis, 

Don M. Nicholson, Anja Boisen, and Thomas Thundat Detection of adsorbed 

explosive molecules using thermal response of suspended microfabricated 

bridges Appl. Phys. Lett. / Volume 93 / Issue 15 16 October 2008 

[254] Gengxin Zhang, Rajasekar Pitchimani, and Brandon L. Weeks A simple and 

flexible thin film evaporating device for energetic materials Rev. Sci. Instrum. / 

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[255] Robert G. Bozic, Alan C. West and Rastislav Levicky Square wave 

voltammetric detection of 2,4,6-trinitrotoluene and 2,4-dinitrotoluene on a gold 

electrode modified with self-assembled monolayers Sensors and Actuators B: 

Chemical Volume 133, Issue 2, 12 August 2008, Pages 509-515 

[256] Toshikazu Kawaguchi, Dhesingh Ravi Shankaran, Sook Jin Kim, Kiyoshi 

Matsumoto, Kiyoshi Toko and Norio Miura. Surface plasmon resonance 

immunosensor using Au nanoparticle for detection of TNT Sensors and 

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M, Akstein Z. Characterization of industrial explosives based on the 

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Litong Jin (2008). Trace detection of nitroaromatic compounds with layer-bylayer 

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B: Chemical Volume 131, Issue 2 Pages 432-438 (May 14) 

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azide and its univalent and divalent ionic forms. Journal of Hazardous Materials 

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screening of biomimetic coatings for selective detection of explosives. 

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detection of explosive residues using photothermal microcantilevers Appl. 

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[264] Hua Bai, Chun Li and Gaoquan Shi (2008) Rapid nitroaromatic compounds 

sensing based on oligopyrene Sensors and Actuators B: Chemical Volume 

130, Issue 2 Pages 777-782 (March 14) 

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electrochemical immunoassay for 2,4,6-trinitrotoluene based on functionalized 

silica nanoparticle labels Analytica Chimica Acta Volume 610, Issue 1 Pages 

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microcantilever using surface reduction. Scanning [Scanning] 2008 Mar-Apr; 

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optimized heat profile Microelectronic Engineering Volume 87, Issues 5-8, 

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fluorescent detector Talanta. 2010 Apr 15;81(1-2):543-8. 

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radiation interrogation to detect explosives Applied Radiation and Isotopes 

Volume 68, Issues 4-5, April-May 2010, Pages 893-895 

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photon signatures Applied Radiation and Isotopes Volume 68, Issues 4-5, 

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[274] Yufang Chen, Zhang Chen, Yejuan He, Hailan Lin, Pengtao Sheng, Chengbin 

Liu, Shenglian Luo and Qingyun Cai L-cysteine-capped CdTe QD-based 

sensor for simple and selective detection of trinitrotoluene Nanotechnology 

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Boisen Micro-calorimetric sensor for vapor phase explosive detection with 

optimized heat profile Microelectronic Engineering Volume 87, Issues 5-8, 

May-August 2010, Pages 696-698 

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radiation interrogation to detect explosives Applied Radiation and Isotopes 

Volume 68, Issues 4-5, April-May 2010, Pages 893-895 

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photon signatures Applied Radiation and Isotopes Volume 68, Issues 4-5, 

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[279] Yufang Chen, Zhang Chen, Yejuan He, Hailan Lin, Pengtao Sheng, Chengbin 

Liu, Shenglian Luo and Qingyun Cai L-cysteine-capped CdTe QD-based 

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[280] Jichang Feng, Yang Li and Mujie Yang Conjugated polymer-grafted silica 

nanoparticles for the sensitive detection of TNT Sensors and Actuators B: 

Chemical Volume 145, Issue 1, 4 March 2010, Pages 438-443 

[281] M Espy, M Flynn, J Gomez, C Hanson, R Kraus, P Magnelind, K Maskaly, A 

Matlashov, S Newman, T Owens, M Peters, H Sandin, I Savukov, L Schultz, A 

Urbaitis, P Volegov and V Zotev Ultra-low-field MRI for the detection of liquid 

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Ray Tsui, Avi Cagan, John Wake, Jeff La Belle and Joseph Wang. Integrated 

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Effect of film thickness, blending and undercoating on optical detection of 

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February 2010, Pages 842-847 

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Margaret M. Timmers Detection of vapor phase trinitrotoluene in the parts-pertrillion 

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Chemical Volume 143, Issue 2, 7 January 2010, Pages 574-582 

[288] Xiangyu Cao, Yasuhiro Oda and Fumihide Shiraishi Photocatalytic and 

adsorptive treatment of 2,4-dinitrophenol using a TiO2 film covering activated 

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[290] Matthew J. Aernecke and David R. Walt Optical-fiber arrays for vapor sensing 

Sensors and Actuators B: Chemical Volume 142, Issue 2, 5 November 2009, 

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detection of trinitrotoluene using off-resonance effects Solid State Nuclear 

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Kiyoshi Matsumoto Development of a sensitive surface plasmon resonance 

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[295] M. Guillemot, F. Dayber, P. Montméat, C. Barthet and P. Prené Detection of 

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A.L. Sanin, I.N. Sorokin, Yu.S. Sulyaev, S.Yu. Taskaev, V.V. Shirokov and 

Yu.I. Eidelman The detection of nitrogen using nuclear resonance absorption 

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[300] Feng Wang, Wenbo Wang, Bianhua Liu, Zhenyang Wang and Zhongping 

Zhang Copolypeptide-doped polyaniline nanofibers for electrochemical 

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[301] Sun Y, Liu J, Frye-Mason G, Ja SJ, Thompson AK, Fan X. Optofluidic ring 

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[302] Yanghai Gui Changsheng Xie, Jiaqiang Xu and Guoqing Wang Detection and 

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[304] Thomas Caron, Marianne Guillemot, Florian Veignal, Pierre Montméat,Eric 

Pasquinet, Philippe Prené, François Perraut, Jean-pierre Lère-Porte, and 

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[333] Jinyou Shen, Jianfa Zhang, Yi Zuo, Lianjun Wang, Xiuyun Sun, Jiansheng Li, 

Weiqing Han and Rui He Biodegradation of 2,4,6-trinitrophenol by 

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Jerald L. Schnoor and Benoit Van Aken Expression of glutathione Stransferases 

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[408] Benoit Van Aken Transgenic plants for enhanced phytoremediation of toxic 

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Jiansheng Li, Weiqing Han and Lu Xu Biodegradation of 2,4,6-trinitrophenol 

(picric acid) in a biological aerated filter (BAF) Bioresource Technology 

Volume 100, Issue 6, March 2009, Pages 1922-1930 

[411] Mary Ann Robinson-Lora and Rachel A. Brennan The use of crab-shell chitin 

for biological denitrification: Batch and column tests Bioresource 

TechnologyVolume 100, Issue 2, January 2009, Pages 534-541 

[412] Ben Stenuit and Spiros N. Agathos Rapid and unbiased colorimetric 

quantification of nitrite and ammonium ions released from 2,4,6-trinitrotoluene 

during biodegradation studies: Eliminating interferences International 

Biodeterioration & Biodegradation Volume 63, Issue 1, January 2009, Pages 

116-122 

[413] Grube M, Muter O, Strikauska S, Gavare M, Limane B. Application of FT-IR 

spectroscopy for control of the medium composition during the biodegradation 

of nitro aromatic compounds. J Ind Microbiol Biotechnol. 2008 

Nov;35(11):1545-9. 

[414] Lihua Li, Jing Wang, Jiti Zhou, Fenglin Yang, Chengyou Jin, Yuanyuan Qu, 

Ang Li and Long Zhang Enhancement of nitroaromatic compounds anaerobic 

biotransformation using a novel immobilized redox mediator prepared by 

electropolymerization Bioresource Technology Volume 99, Issue 15, October 

2008, Pages 6908-6916 

[415] Zlatka Alexieva, Maria Gerginova, Plamena Zlateva, Jordan Manasiev, Danka 

Ivanova and Nely Dimova (2008) Monitoring of aromatic pollutants 

biodegradation Biochemical Engineering Journal Volume 40, Issue 2 Pages 

233-240 

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[416] Ziganshin, A. M.; Naumov, A. V.; Suvorova, E. S.; Naumenko, E. A.; Naumova, 

R. P Hydride-mediated reduction of 2,4,6-trinitrotoluene by yeasts as the way 

to its deep degradation. Microbiology (00262617), Nov2007, Vol. 76 Issue 6, 

p676-682 

[417] Jackson RG, Rylott EL, Fournier D, Hawari J, Bruce NC. (2007) Exploring the 

biochemical properties and remediation applications of the unusual explosivedegrading 

P450 system XplA/B Proc Natl Acad Sci U S A. 104(43):16822-7. 

  

[418] Zhao JS, Manno D, Thiboutot S, Ampleman G, Hawari J. (2007) Shewanella 

canadensis sp. nov. and Shewanella atlantica sp. nov., manganese dioxide- 

and hexahydro-1,3,5-trinitro-1,3,5-triazine-reducing, psychrophilic marine 

bacteria. Int J Syst Evol Microbiol Sep;57(Pt 9):2155-62 

  

New, Novel Explosives; Synthesis and Characterization: Properties, 

Sensitivity, Kinetics, Detonation Properties, Degredation 

[419] Byoung Sun Min and Young Chul Park A study on the aliphatic energetic 

plasticizers containing nitrate ester and nitramine Journal of Industrial and 

Engineering Chemistry Volume 15, Issue 4, 25 July 2009, Pages 595-601 

[420] Lewis L. Stevens, Nenad Velisavljevic, Daniel E. Hooks, and Dana M. 

Dattelbaum The high-pressure phase behavior and compressibility of 2,4,6trinitrotoluene 

Appl. Phys. Lett. / Volume 93 / Issue 8 081912 27 August 2008 

[421] Xiufang Ma, Weihua Zhu, Jijun Xiao and Heming Xiao Molecular dynamics 

study of the structure and performance of simple and double bases propellants 

Journal of Hazardous Materials Volume 156, Issues 1-3, 15 August 2008, 

Pages 201-207 

[422] Alex M. Remennikov and Timothy A. Rose (2007) Predicting the effectiveness 

of blast wall barriers using neural networks International Journal of Impact 

Engineering Volume 34, Issue 12, Pages 1907-1923 

[423] Huaxiong Chen, Shusen Chen, Lijie Li, Qingze Jiao, Tianyu Wei and Shaohua 

Jin Synthesis, single crystal structure and characterization of 

pentanitromonoformylhexaazaisowurtzitane Journal of Hazardous Materials 

Volume 175, Issues 1-3, 15 March 2010, Pages 569-574 

[424] A.Saikia, R. Sivabalan, B.G. Polke, G.M. Gore, Amarjit Singh, A. Subhananda 

Rao and A.K. Sikder Synthesis and characterization of 3,6-bis(1H-1,2,3,4tetrazol-5-ylamino)-1,2,4,5-tetrazine 

(BTATz): Novel high-nitrogen content 

insensitive high energy material Journal of Hazardous Materials Volume 170, 

Issue 1, 15 October 2009, Pages 306-313 

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[425] Hongyan Chen, Tonglai Zhang and Jianguo Zhang Synthesis, characterization 

and properties of nitrogen-rich salts of trinitrophloroglucinol Journal of 

Hazardous Materials Volume 161, Issues 2-3, 30 January 2009, Pages 1473- 

1477 

[426] Thomas M. Klapötke, Norma K. Minar and Jörg Stierstorfer Investigations of 

bis(methyltetrazolyl)triazenes as nitrogen-rich ingredients in solid rocket 

propellants – Synthesis, characterization and properties Polyhedron Volume 

28, Issue 1, 14 January 2009, Pages 13-26 

[427] Sheng Jun Luo, Wan Long Pan, Ying Nan Chi, Yan Qing Xu, Kun Lin Huan and 

Chang Wen Hu Synthesis and characterization of a novel high thermally stable 

energy compound: 1-(1-Adamantylamino)-2,4,6-trinitrobenzene Chinese 

Chemical Letters Volume 19, Issue 10, October 2008, Pages 1147-1150 

[428] Liqiong Wang, Hongyan Chen, Tonglai Zhang, Jianguo Zhang and Li Yang 

(2007) Synthesis, characterization, thermal and explosive properties of 

potassium salts of trinitrophloroglucinol Journal of Hazardous Materials Volume 

147, Issues 1-2, Pages 576-580 

[429] Victor J. Bellitto and Mikhail I. Melnik Surface defects and their role in the 

shock sensitivity of cyclotrimethylene-trinitramine Applied Surface Science 

Volume 256, Issue 11, 15 March 2010, Pages 3478-3481 

[430] Jinshan Li A multivariate relationship for the impact sensitivities of energetic Nnitrocompounds 

based on bond dissociation energy Journal of Hazardous 

Materials Volume 174, Issues 1-3, 15 February 2010, Pages 728-733 

[431] Lemi Türker Contemplation on spark sensitivity of certain nitramine type 

explosives Journal of Hazardous Materials Volume 169, Issues 1-3, 30 

September 2009, Pages 454-459 

[432] Lemi Türker Tunneling effect and impact sensitivity of certain explosives 

Journal of Hazardous Materials Volume 169, Issues 1-3, 30 September 2009, 

Pages 819-823 

[433] Mohammad Hossein Keshavar , Hamid Reza Pouretedal and Abolfazl Semnani 

Reliable prediction of electric spark sensitivity of nitramines: A general 

correlation with detonation pressure Journal of Hazardous Materials Volume 

167, Issues 1-3, 15 August 2009, Pages 461-466 

[434] Keshavarz, Mohammad Hossein; Zali, Abbas; Shokrolahi, Arash A simple 

approach for predicting impact sensitivity of polynitroheteroarenes Journal of 

Hazardous Materials, Jul2009, Vol. 166 Issue 2/3, p1115-1119 

  

  

  

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[435] Xiaolan Song, Yi Wang, Chongwei An, Xiaode Guo and Fengsheng Li 

Dependence of particle morphology and size on the mechanical sensitivity and 

thermal stability of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine Journal of 

Hazardous Materials Volume 159, Issues 2-3, 30 November 2008, Pages 222- 

229 

[436] V.P. Sinditskii, M.C. Vu, A.B. Sheremetev and N.S. Alexandrova Study on 

thermal decomposition and combustion of insensitive explosive 3,3′-diamino- 

4,4′-azofurazan (DAAzF) Thermochimica Acta Volume 473, Issues 1-2, 10 

July 2008, Pages 25-31 

[437] Xiaoshu Song, Xinlu Cheng, Xiangdong Yang, Dehua Li and Rongfeng Linghu 

Correlation between the bond dissociation energies and impact sensitivities in 

nitramine and polynitro benzoate molecules with polynitro alkyl groupings 

Journal of Hazardous Materials Volume 150, Issue 2, 31 January 2008, Pages 

317-321 

[438] H. Czerski and W.G. Proud. Relationship between the morphology of granular 

cyclotrimethylene-trinitramine and its shock sensitivity. Journal of Applied 

Physics 102.11 (Dec 1, 2007): p.113515-1-113515-8 

  

[439] Peterson, P. D.; Lee, K.-Y.; Moore, D. S.; Scharff, R. J.; Avilucea The Evolution 

Of Sensitivity In HMX-Based Explosives During The Reversion From Delta To 

Beta Phase. AIP Conference Proceedings, 12/12/2007, Vol. 955 Issue 1, p987- 

990 

  

[440] V. Dubovik Analysis of standard indices of impact sensitivity of solid explosives 

Doklady Physical Chemistry 415.2 (August 2007): p.218(3). R. K. Weese, A. K. 

Burnham, Heidi C. Turner and T. D. Tran. Exploring the physical, chemical and 

thermal characteristics of a new potentially insensitive high explosive RX-55- 

AE-5 Journal of Thermal Analysis and Calorimetry 89.2 (August 


  

[441] R. K. Weese, A. K. Burnham, Heidi C. Turner and T. D. Tran. Exploring the 

physical, chemical and thermal characteristics of a new potentially insensitive 

high explosive RX-55-AE-5 Journal of Thermal Analysis and 

Calorimetry 89.2 (August 2007): p.465(9). 

  

[442] M.R. Sovizi, S.S. Hajimirsadeghi and B. Naderizadeh Effect of particle size on 

thermal decomposition of nitrocellulose Journal of Hazardous Materials 

Volume 168, Issues 2-3, 15 September 2009, Pages 1134-1139 

[443] I.Häring, M. Schönherr and C. Richter Quantitative hazard and risk analysis for 

fragments of high-explosive shells in air Reliability Engineering & System 

Safety Volume 94, Issue 9, September 2009, Pages 1461-1470 

510

[444] Xinghua Xie , Jing Zhu and Huisheng Zhou Deflagration products from 

emulsion explosive unconventional nano-powders from ion reaction Journal of 

Physics: Conference Series Volume 188, Number 1 p. 012003 

[445] Z.W. Liu, H.M. Xie, K.X. Li, P.W. Chen and F.L. Huang Fracture behavior of 

PBX simulation subject to combined thermal and mechanical loads Polymer 

Testing Volume 28, Issue 6, September 2009, Pages 627-635 

[446] Jennifer Mott Peuker, Patrick Lynch, Herman Krier and Nick Glumac Optical 

depth measurements of fireballs from aluminized high explosives Optics and 

Lasers in Engineering Volume 47, Issue 9, September 2009, Pages 1009-1015 

[447] Yu, Weifei; Zhang, Tonglai; Huang, Yigang; Yang, Li; Li, Gang; Li, Haibo; Li, 

Jinshan; Huang, Hui. Effect of microwave irradiation on TATB explosive. 

Journal of Hazardous Materials, Sep2009, Vol. 168 Issue 2/3, p952-954 

  

[448] Song Guo, Qingsong Wang, Jinhua Sun , , Xin Liao and Ze-shan Wang Study 

on the influence of moisture content on thermal stability of propellant Journal 

of Hazardous Materials Volume 168, Issue 1, 30 August 2009, Pages 536-541 

[449] R.S. Damse, B. Omprakash, B.G. Tope, T.K. Chakraborthy and Amarjit Singh 

Study of N-n-butyl-N-(2-nitroxyethyl)nitramine in RDX based gun propellant 

Journal of Hazardous Materials Volume 167, Issues 1-3, 15 August 2009, 

Pages 1222-1225 

[450] Zhang Jiao-qiang, Gao Hong-xu, Su Li-hong, Hu Rong-zu, Zhao Feng-qi and 

Wang Bo-zhou Non-isothermal thermal decomposition reaction kinetics of 2nitroimino-5-nitro-hexahydro-1,3,5-triazine 

(NNHT) Journal of Hazardous 

Materials Volume 167, Issues 1-3, 15 August 2009, Pages 205-208 

[451] Keshavarz, Mohammad Hossein Simple correlation for predicting detonation 

velocity of ideal and non-ideal explosives. Journal of Hazardous Materials, 

Jul2009, Vol. 166 Issue 2/3, p762-769 

  

[452] Limei Qiu, Xuedong Gong, Jian Zheng and Heming Xiao Theoretical studies 

on polynitro-1,3-bishomopentaprismanes as potential high energy density 

compounds. Journal of Hazardous Materials Volume 166, Issues 2-3, 30 July 

2009, Pages 931-938 

  

[453] R.S. Damse and A.K. Sikder Suitability of nitrogen rich compounds for gun 

propellant formulations Journal of Hazardous Materials Volume 166, Issues 2- 

3, 30 July 2009, Pages 967-971 

  

[454] Keshavarz, Mohammad Hossein Prediction of detonation performance of 

CHNO and CHNOAl explosives through molecular structure Journal of 

Hazardous Materials, Jul2009, Vol. 166 Issue 2/3, p1296-1301 

  

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[455] Dmitry Bedrov, Justin B. Hooper, Grant D. Smith and Thomas D. Sewell 

Shock-induced transformations in crystalline RDX: A uniaxial constant-stress 

Hugoniostat molecular dynamics simulation study J. Chem. Phys. 131, 034712 

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[456] Im, Hee-Jung; Song, Byoung Chul; Park, Yong Joon; Song, Kyuseok 

Classification of materials for explosives from prompt gamma spectra by using 

principal component analysis. Applied Radiation & Isotopes, Jul2009, Vol. 67 

Issue 7/8, p1458-1462 

  

[457] Xin Zhang, Yu-man Lin and Zu-liang Chen 2,4,6-Trinitrotoluene reduction 

kinetics in aqueous solution using nanoscale zero-valent iron Journal of 

Hazardous Materials Volume 165, Issues 1-3, 15 June 2009, Pages 923-927 

[458] Jian-hua Yi, Feng-qi Zhao, Si-yu Xu, La-ying Zhang, Hong-xu Gao and Rongzu 

Hu Effects of pressure and TEGDN content on decomposition reaction 

mechanism and kinetics of DB gun propellant containing the mixed ester of 

TEGDN and NG Journal of Hazardous Materials Volume 165, Issues 1-3, 15 

June 2009, Pages 853-859 

[459] Richard Gunawan and Dongke Zhang Thermal stability and kinetics of 

decomposition of ammonium nitrate in the presence of pyrite Journal of 

Hazardous Materials Volume 165, Issues 1-3, 15 June 2009, Pages 751-758 

[460] Xiaohong Li, Zhengxin Tang, Xianzhou Zhang and Xiangdong Yang The heats 

of formation in a series of nitroester energetic compounds: A theoretical study 

Journal of Hazardous Materials Volume 165, Issues 1-3, 15 June 2009, Pages 

372-378 

[461] Ryan M. Richard and David W. Ball. B3LYP calculations on the 

thermodynamic properties of a series of nitroxycubanes having the formula 

C8H8−x(NO3)x (x = 1–8) Journal of Hazardous Materials Volume 164, Issues 2- 

3, 30 May 2009, Pages 1595-1600 

[462] Dušan Klasovitý, Svatopluk Zeman , , Aleš Růžička, Marcela Jungová and 

Michal Roháč cis-1,3,4,6-Tetranitrooctahydroimidazo-[4,5-d]imidazole 

(BCHMX), its properties and initiation reactivity Journal of Hazardous Materials 

Volume 164, Issues 2-3, 30 May 2009, Pages 954-961 

[463] Wei Zhu Jijun Xiao, Weihua Zhu and Heming Xiao Molecular dynamics 

simulations of RDX and RDX-based plastic-bonded explosives Journal of 

Hazardous Materials Volume 164, Issues 2-3, 30 May 2009, Pages 1082-1088 

[464] Shyam S. Shukla, Kenneth L. Dorris and Bhaskara V. Chikkaveeraiah 

Photocatalytic degradation of 2,4-dinitrophenol Journal of Hazardous Materials 

Volume 164, Issue 1, 15 May 2009, Pages 310-314 

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[465] Ling Qiu and Heming Xiao Molecular dynamics study of binding energies, 

mechanical properties, and detonation performances of bicyclo-HMX-based 

PBXs Journal of Hazardous Materials Volume 164, Issue 1, 15 May 2009, 

Pages 329-336 

[466] Mauracher, S. Denifl, M. Probst, T. D. Märk, and P. Scheier Investigations Of 

Electron Attachment To Nitro-Compounds Towards Explosives AIP Conf. 

Proc. -- May 2, 2009 -- Volume 1125, pp. 157-165 

[467] Marc J. Schniederjans, Michelle L. Pantoya, James J. Hoffman and Darrin L. 

Willauer A multi-objective modeling approach for energetic material evaluation 

decisions European Journal of Operational Research Volume 194, Issue 3, 1 

May 2009, Pages 629-636 

[468] Jacques Bouillard, Alexis Vignes, Olivier Dufaud, Laurent Perrin and 

Dominique Thomas Explosion risks from nanomaterials Journal of Physics: 

Conference Series Volume 170, Number 1 012032 

[469] Miao-miao Li, Gui-xiang Wang, Xiao-de Guo, Zhan-wu Wu and Hong-chang 

Song Theoretical studies on the structures, thermodynamic properties, 

detonation properties, and pyrolysis mechanisms of four trinitrate esters 

Journal of Molecular Structure: THEOCHEM Volume 900, Issues 1-3, 30 April 

2009, Pages 90-95 

[470] Weihua Zhu, Xiaowen Zhang, Tao Wei and Heming Xiao First-principles study 

of crystalline mono-amino-2,4,6-trinitrobenzene, 1,3-diamino-2,4,6trinitrobenzene, 

and 1,3,5-triamino-2,4,6-trinitrobenzene Journal of Molecular 

Structure: THEOCHEM Volume 900, Issues 1-3, 30 April 2009, Pages 84-89 

[471] C. Gruau, D. Picart, R. Belmas, E. Bouton, F. Delmaire-Sizes, J. Sabatier and 

H. Trumel Ignition of a confined high explosive under low velocity impact 

International Journal of Impact Engineering Volume 36, Issue 4, April 2009, 

Pages 537-550 

[472] Alexey Makarov, Rosario LoBrutto, Christos Christodoulatos and Anton 

Jerkovich The use of ultra high-performance liquid chromatography for 

studying hydrolysis kinetics of CL-20 and related energetic compounds Journal 

of Hazardous Materials Volume 162, Issues 2-3, 15 March 2009, Pages 1034- 

1040 

[473] T. Mondal, B. Saritha, S. Ghanta, T.K. Roy, S. Mahapatra and M. Durga 

Prasad On some strategies to design new high energy density molecules 

Journal of Molecular Structure: THEOCHEM Volume 897, Issues 1-3, 15 March 

2009, Pages 42-47 

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[474] Mohammad Hossein Keshavarz Simple method for prediction of activation 

energies of the thermal decomposition of nitramines Journal of Hazardous 

Materials Volume 162, Issues 2-3, 15 March 2009, Pages 1557-1562 

[475] S.M. Pourmortazavi, S.G. Hosseini, M. Rahimi-Nasrabadi, S.S. Hajimirsadeghi 

and H. Momenian Effect of nitrate content on thermal decomposition of 

nitrocellulose Journal of Hazardous Materials Volume 162, Issues 2-3, 15 

March 2009, Pages 1141-1144 

[476] Yingxin Zhou and Arnfinn Jenssen Internal separation distances for 

underground explosives storage in hard rock Tunnelling and Underground 

Space Technology Volume 24, Issue 2, March 2009, Pages 119-125 

[477] Mi-An Xue, Xiaoli Yuan, Xiaoying Fu, Jinghe Wu, Pengzhi Lin, Dong Hu, Song 

Ye, Wenjun Zhu, Yanping Wang and Zhongwei Huang Effects of copper 

micro-particles on the detonation characteristics of RDX powder J. Phys. D: 

Appl. Phys. 42:4 p. 045504 Issue 4 (21 February 2009) 

[478] Lemi Türker and Taner Atalar Computational studies on nitratoethylnitramine 

(NENA), its tautomers and charged forms Journal of Hazardous Materials 

Volume 162, Issue 1, 15 February 2009, Pages 193-203 

[479] Weihua Zhu, Tao Wei, Xiaowen Zhang and Heming Xiao Density functional 

theory study of structural, vibrational, and thermodynamic properties of 

crystalline 2,4-dinitrophenol, 2,4-dinitroresorcinol, and 4,6-dinitroresorcinol 

Journal of Molecular Structure: THEOCHEM Volume 895, Issues 1-3, 15 

February 2009, Pages 131-137 

[480] Gregory Young, Kyle Sullivan, Michael R. Zachariah and Kenneth Yu 

Combustion characteristics of boron nanoparticles Combustion and Flame 


[481] Xiu-E Shen, Xiao-Quan Shan, De-Ming Dong, Xiu-Yi Hua and Gary Owens 

Kinetics and thermodynamics of sorption of nitroaromatic compounds to asgrown 

and oxidized multiwalled carbon nanotubes Journal of Colloid and 

Interface Science Volume 330, Issue 1, 1 February 2009, Pages 1-8 

[482] Leonid Kagan and Gregory Sivashinsky An elementary model for the transition 

from conductive to penetrative burning in gas-permeable explosives 

Combustion and Flame Volume 156, Issue 2, February 2009, Pages 531-538 

[483] R.J. Kalbasi, M. Ghiaci and A.R. Massah Highly selective vapor phase 

nitration of toluene to 4-nitro toluene using modified and unmodified 

H3PO4/ZSM-5 Applied Catalysis A: General Volume 353, Issue 1, 31 January 

2009, Pages 1-8 

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[484] H. Muthurajan, R. Sivabalan, N. Pon Saravanan and M.B. Talawar Computer 

code to predict the heat of explosion of high energy materials Journal of 

Hazardous Materials Volume 161, Issues 2-3, 30 January 2009, Pages 714- 

717 

[485] Xinfang Su, Xinlu Cheng, Chuanmin Meng and Xiaoli Yuan Quantum chemical 

study on nitroimidazole, polynitroimidazole and their methyl derivatives Journal 

of Hazardous Materials Volume 161, Issue 1, 15 January 2009, Pages 551-558 

[486] Fengbin Lei, Zuoshan Wang and Baoguo Zhao Preparation and mechanism of 

nanometer Al5O6N via shock wave plasma technique Materials Research 

Bulletin Volume 44, Issue 1, 8 January 2009, Pages 198-201 

[487] Yana Kholod, Dmytro Kosenkov, Sergiy Okovytyy, Leonid Gorb, Mohammad 

Qasim and Jerzy Leszczynsk CL-20 photodecomposition: Ab initio foundations 

for identification of products Spectrochimica Acta Part A: Molecular and 

Biomolecular Spectroscopy Volume 71, Issue 1, 1 November 2008, Pages 230- 

237 

[488] Hai-Xia Ma, Ji-Rong Song, Rong-Zu Hu, Qing Pan and Yuan Wang Nonisothermal 

decomposition kinetics, thermal behavior and computational 

detonation properties on 4-amino-1,2,4-triazol-5-one (ATO) Journal of 

Analytical and Applied Pyrolysis Volume 83, Issue 2, November 2008, Pages 

145-150 

[489] Seok-Young Oh, Pei C. Chiu and Daniel K. Cha Reductive transformation of 

2,4,6-trinitrotoluene, hexahydro-1,3,5-trinitro-1,3,5-triazine, and nitroglycerin by 

pyrite and magnetite Journal of Hazardous Materials Volume 158, Issues 2-3, 

30 October 2008, Pages 652-655 

[490] Sylke G. Boyd and Kevin J. Boyd A computational analysis of the interaction 

of lattice and intramolecular vibrational modes in crystalline -RDX J. Chem. 

Phys. / Volume 129 / Issue 13 1 October 2008 

[492] Andrew J. Heim, Niels Grønbech-Jensen, Edward M. Kober, Jerome J. 

Erpenbeck, and Timothy C. Germann Interaction potential for atomic 

simulations of conventional high explosives Phys. Rev. E Volume 78 Issue 4 

(October 28, 2008) 

[493] Xiao-Wei Fan, Xue-Hai Ju and He-Ming Xiao Density functional theory study of 

piperidine and diazocine compounds Journal of Hazardous Materials Volume 

156, Issues 1-3, 15 August 2008, Pages 342-347 

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[494] S. Jönsson, L.A. Eriksson and B. van Bavel Multivariate characterisation and 

quantitative structure–property relationship modelling of nitroaromatic 

compounds Analytica Chimica Acta Volume 621, Issue 2, 28 July 2008, Pages 

155-162 

[495] M Comet, B Siegert, V Pichot, P Gibot and D Spitzer Preparation of explosive 

nanoparticles in a porous chromium(III) oxide matrix: a first attempt to control 

the reactivity of explosives Nanotechnology 19 285716 

[496] J.A. Sanchidrián, L.M. López and P. Segarra The influence of some blasting 

techniques on the probability of ignition of firedamp by permissible explosives 

Journal of Hazardous Materials Volume 155, Issue 3, 15 July 2008, Pages 580- 

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[497] Kenneth J. Balkus Jr., Thomas J. Pisklak, Greg Hundt, John Sibert and 

Yangfeng Zhang (2008) Photoluminescent and redox active periodic 

mesoporous organosilicas based on 2,7-diazapyrene Microporous and 

Mesoporous Materials Volume 112, Issues 1-3, Pages 1-13 (July) 

[498] Ulrich Hauptmanns (2008) Comparative assessment of the dynamic behaviour 

of an exothermal chemical reaction including data uncertainties Chemical 

Engineering Journal Volume 140, Issues 1-3, Pages 278-286 

[499] Yunlan Sun and Shufen Li (2008) The effect of nitrate esters on the thermal 

decomposition mechanism of GAP Journal of Hazardous Materials Volume 

154, Issues 1-3, Pages 112-117 (June 15) 

[500] Zhenhua Liu, Tonglai Zhang, Jianguo Zhang and Shaozong Wang (2008) 

Studies on three-dimensional coordination polymer [Cd2(N2H4)2(N3)4]n: Crystal 

structure, thermal decomposition mechanism and explosive properties Journal 

of Hazardous Materials Volume 154, Issues 1-3 Pages 832-838 

[501] Martin Rahm and Tore Brinck (2008) Dinitraminic acid (HDN) isomerization and 

self-decomposition revisited Chemical Physics Volume 348, Issues 1-3, Pages 

53-60 

[502] Su-hong Ge, Xin-lu Cheng, Zheng-lai Liu, Xiang-dong Yang and Fang-fang 

Dong Correlation Between Energy Transfer Rate and Atomization Energy of 

Some Trinitro Aromatic Explosive Molecules Chin. J. Chem. Phys. 21:3 250- 

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[503] Wen-Shing Chenand Jing-Song Liang (2008) Decomposition of nitrotoluenes 

from trinitrotoluene manufacturing process by Electro-Fenton oxidation 

Chemosphere Volume 72, Issue 4, Pages 601-607 

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[504] Mrinal Kanti Mandal, Sukalyan Dutta and P.K. Bhattacharya (2008) 

Characterization of blended polymeric membranes for pervaporation of 

hydrazine hydrate Chemical Engineering Journal Volume 138, Issues 1-3, 

Pages 10-19 

[505] Wen-Shing Chenand Guo-Chao Huang (2008) Ultrasound-assisted 

crystallization of high purity of 2,4-dinitrotoluene from spent acid Ultrasonics 

Sonochemistry Volume 15, Issue 5, Pages 909-915 (May 2008) 

  

[506] Hubertus Giefers and Michael Pravica. Radiation-induced decomposition of 

PETN and TATB under extreme conditions. Journal of Physical Chemistry 

A 112.15 (April 17, 2008): p.3352-3359. 

  

[507] D M Williamson, C R Siviour, W G Proud, S J P Palmer, R Govier, K Ellis, P 

Blackwell and C Leppard (2008) Temperature–time response of a polymer 

bonded explosive in compression (EDC37) J. Phys. D: Appl. Phys. 41:8 

085404 (10pp) (April 21) 

[508] Kyung-cheol Lee, Ki-hong Kim, and Jack J. Yoh (2008) Modeling of high 

energy laser ignition of energetic materials J. Appl. Phys. / Volume 103 / Issue 

8 / 083536 

[510] Xiao-Hong Li, Zheng-Xin Tang, Abraham F. Jalbout, Xian-Zhou Zhang and Xin- 

Lu Cheng (2008) A DFT study of bond dissociation energies of several alkyl 

nitrate and nitrite compounds Journal of Molecular Structure: THEOCHEM 

Volume 854, Issues 1-3, Pages 76-80 

[511] Staiano M; Saviano M; Herman P; Grycznyski Z; Fini C; Varriale A; Parracino 

A; Kold AB; Rossi M; D'Auria S Time-resolved fluorescence spectroscopy and 

molecular dynamics simulations point out the effects of pressure on the stability 

and dynamics of the porcine odorant-binding protein. Biopolymers 

[Biopolymers] 2008 Apr; Vol. 89 (4), pp. 284-91 

[512] Badgujar, D.M. Talawar, M.B. Asthana, S.N. Mahulikar, P.P. Advances in 

science and technology of modern energetic materials: An overview Journal of 

Hazardous Materials; Mar2008, Vol. 151 Issue 2/3, p289-305 

[513] Qi-Long Yan, Xiao-Jiang Li, Han Wang, Li-Hua Nie, Ze-Yi Zhang and Hong-Xu 

Gao Thermal decomposition and kinetics studies on 1,4-dinitropiperazine 

(DNP) Journal of Hazardous Materials Volume 151, Issues 2-3, 1 March 2008, 

Pages 515-521 

  

[514] Xiufang Ma, Feng Zhao, Guangfu Ji, Weihua Zhu, Jijun Xiao and Heming Xiao 

(2008) Computational study of structure and performance of four constituents 

HMX-based composite material Journal of Molecular Structure: THEOCHEM 

Volume 851, Issues 1-3 Pages 22-29 

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[515] Jijun Xiao, Hui Huang, Jinshan Li, Hang Zhang, Wei Zhu and Heming Xiao 

(2008) A molecular dynamics study of interface interactions and mechanical 

properties of HMX-based PBXs with PEG and HTPB Journal of Molecular 

Structure: THEOCHEM Volume 851, Issues 1-3, Pages 242-248 

[516] Inder Pal Singh Kapoor, Pratibha Srivastava and Gurdip Singh (2008) 

Preparation, characterization and thermolysis of phenylenediammonium 

dinitrate salts Journal of Hazardous Materials Volume 150, Issue 3, Pages 687- 

694 (Feb 11) 

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Charge-density studies of energetic materials: CL-20 and FOX-7 Acta 

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[531] Martina Chovancova and Svatopluk Zeman Study of initiation reactivity of some 

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[534] R. Pitchimani, W. Zheng, S. L. Simon, L. J. Hope-Weeks, A. K. 

Burnham and B. L. Weeks Thermodynamic analysis of pure and impurity 

doped pentaerythritol tetranitrate crystals grown at room temperature. Journal 

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[535] Chanyang Zhang. Computational investigation on the desensitizing mechanism 

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Journal of Physical Chemistry B 111.22 (June 7, 2007): p.6208-6213. 

  

[536] D.L. Frost, C. Ornthanalai, Z. Zarei, V. Tanguay and F. Zhang Particle 

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[537] Merrill W. Beckstead, Karthik Puduppakkam, Piyush Thakre and Vigor Yang 

(2007) Modeling of combustion and ignition of solid-propellant ingredients 

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[538] Ming-Jer Liou and Ming-Chun Lu (2007) Catalytic degradation of nitroaromatic 

explosives with Fenton's reagent Journal of Molecular Catalysis A: Chemical 


[539] G.A. Pinhasi, A. Ullmann and A. Dayan (2007) 1D plane numerical model for 

boiling liquid expanding vapor explosion (BLEVE) International Journal of Heat 

and Mass Transfer Volume 50, Issues 23-24 Pages 4780-4795 

[540] Yunfeng Shi and Donald W. Brenner (2007) Simulated thermal decomposition 

and detonation of nitrogen cubane by molecular dynamics J. Chem. Phys. 127 

134503-134509 

[541] Antoine Osmont, Laurent Catoire, Iskender Gökalp and Vigor Yang (2007). Ab 

initio quantum chemical predictions of enthalpies of formation, heat capacities, 

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[542] Luísa Durães, Benilde F.O. Costa, Regina Santos, António Correia, José 

Campos and António Portugal (2007) Fe2O3/aluminum thermite reaction 

intermediate and final products characterization Materials Science and 

Engineering: A Volume 465, Issues 1-2 Pages 199-210 

[543] Martina Chovancová and Svatopluk Zeman (2007) Study of initiation reactivity 

of some plastic explosives by vacuum stability test and non-isothermal 

differential thermal analysis Thermochimica Acta Volume 460, Issues 1-2, 

Pages 67-76 

[544] S. Ravi Annapragada, Dawei Sun and Suresh V. Garimella (2007) Prediction of 

effective thermo-mechanical properties of particulate composites 

Computational Materials Science Volume 40, Issue 2, Pages 255-266 

[545] Gui-xiang Wang, Xue-dong Gong, Yan Liu and He-ming Xiao A theoretical 

study on the vibrational spectra and thermodynamic properties for the nitro 

derivatives of phenols Spectrochimica Acta Part A: Molecular and 

Biomolecular Spectroscopy Volume 74, Issue 2, 1 October 2009, Pages 569- 

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[546] Mohammad Hossein Keshavarz and Hamid Reza Pouretedal A reliable simple 

method to estimate density of nitroaliphatics, nitrate esters and nitramines 

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Pages 158-169 

LandMine, Mine Detection, Demining 

[547] Corcelli A, Lobasso S, Lopalco P, Dibattista M, Araneda R, Peterlin Z, Firestein 

S. Detection of explosives by olfactory sensory neurons. J Hazard Mater. 

2010 Mar 15;175(1-3):1096-100. 

[548] Victor Bom, A.M. Osman and A.M. Abdel Monem A novel scanning land mine 

detector based on the technique of neutron back scattering imaging IEEE 

Transactions on Nuclear Science 55.2 (April 2008): p.741(7). 

  

[549] E.T.H. Clifford, J.E. McFee, H. Ing, H.R. Andrews, D. Tennant, E. Harper and 

A.A. Faust (2007) A militarily fielded thermal neutron activation sensor for 

landmine detection Nuclear Instruments and Methods in Physics Research 

Section A: Accelerators, Spectrometers, Detectors and Associated Equipment 

Volume 579, Issue 1 Pages 418-425 

[550] D. Rezaei Ochbelagh, H. Miri Hakimabad and R. Izadi Najafabadi (2007) The 

investigation of Am–Be neutron source shield effect used on landmine 

detection Nuclear Instruments and Methods in Physics Research Section A: 


577, Issue 3 Pages 756-761 

[551] Maik Irrazábal, Samuel P. Hernández-Rivera and Julio G. Briano Modeling of 

TNT transport from landmines: Numerical approach Chemosphere Volume 77, 

Issue 4, October 2009, Pages 546-551 

[552] Hashem Miri-Hakimabad and Alireza Vejdani-Noghreiyan Increasing the 

Personnel Safety of a Landmine Detection System using Proper Shielding 

Materials Journal of Medical Imaging and Radiation Sciences Volume 40, 

Issue 3, September 2009, Pages 105-108 

[553] Zare, Alina; Bolton, Jeremy; Gader, Paul; Schatten, Miranda Vegetation 

Mapping for Landmine Detection Using Long-Wave Hyperspectral Imagery. 

IEEE Transactions on Geoscience & Remote Sensing, Jan2008, Vol. 46 Issue 

1, p172-178, 

  

[554] Maki K. Habib Controlled biological and biomimetic systems for landmine 

detection. Biosensors and Bioelectronics 23.1 (August 30, 2007): p.1(18). 

  

[555] Lihan He; Shihao Ji; Scott Jr., Waymond R.; Carin, Lawrence Adaptive 

Multimodality Sensing of Landmines. IEEE Transactions on Geoscience & 

Remote Sensing, Jun2007 Part 2, Vol. 45, p1756-1774, 

  

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[556] Maki K. Habib (2007) Controlled biological and biomimetic systems for 

landmine detection Biosensors and Bioelectronics Volume 23, Issue 1, Pages 

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[557] Grujicic, M.; Pandurangan, B.; Summers, J.D.; Cheeseman, B.A.; Roy, W.N.; 

Skaggs, R.R Application of the modified compaction material model to the 

analysis of landmine detonation in soil with various degrees of water saturation. 

Shock & Vibration, 2008, Vol. 15 Issue 1, p79-99, 

  

[558] Olga Lopera and Nada Milisavljevic (2007) Prediction of the effects of soil and 

target properties on the antipersonnel landmine detection performance of 

ground-penetrating radar: A Colombian case study Journal of Applied 

Geophysics Volume 63, Issue 1, Pages 13-23 

[559] N.; Skaggs, R. R.. A Combined Multi-Material Euler/Lagrange Computational 

Analysis of Blast Loading Resulting from Detonation of Buried Landmines. 

Multidiscipline Modeling in Materials & Structures, 2008, Vol. 4 Issue 2, p105- 

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[560] McFee, J. E.; Faust, A. A.; Andrews, H. R.; Kovaltchouk, V.; Clifford, E. T.; Ing, 

H A Comparison of Fast Inorganic Scintillators for Thermal Neutron Analysis 

Landmine Detection. Transactions on Nuclear Science, Jun2009 Part 3 of 3, 

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[561] Yoshikawa, Kiyoshi; Masuda, Kai; Takamatsu, Teruhisa; Yamamoto, Yasushi; 

Toku, Hisayuki; Fujimoto, Takeshi; Hotta, Eiki; Yamauchi, Kunihito; Ohnishi, 

Masami; Osawa, Hodaka; Shiroya, Seiji; Misawa, Tsuyoshi; Takahashi, 

Yoshiyuki; Kubo, Yoshikazu; Doi, Toshiro Research and Development of the 

Humanitarian Landmine Detection System by a Compact Fusion Neutron 

Source. IEEE Transactions on Nuclear Science, Jun2009 Part 2 of 3, Vol. 56 

Issue 3, p1193-1202 

  

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Hisayuki Toku, Takashi Fujimoto, Eiki Hotta, Kunihito Yamauchi, Masami 

Ohnishi, Hodaka Osawa, Seiji Shiroya, Tsuyoshi Misawa, Yoshiyuki Takahashi, 

Yoshikazu Kubo, and Toshiro Doi Results of the Development of Humanitarian 

Landmine Detection System by a Compact Fusion Neutron Source and Dual 

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Explosives Safety 

[563] Tasneem Abbasi, H.J. Pasman and S.A. Abbasi A scheme for the 

classification of explosions in the chemical process industry Journal of 

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280 

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523

Hazardous materials: 

Chemical biological radiological 

And nuclear 


Eric Wenger, PhD 

Australian CBRN Data Centre 

Australian Federal Police 

GPO Box 401 

Canberra, ACT 2601 

Australia 

524


1.  EXECUTIVE SUMMARY  526  


3.  CURRENT CBRN TRENDS  527  

3.1  Chemical  528  

3.2  Biological  528  

3.3  Radiological and nuclear  528  

4.  DETECTION  528  

4.1  Field Detection  529  

4.1.2 Chemical  529  

4.1.3  Biological  530  

4.1.4  Radiological  532  

4.2  OpticallyStimulated Luminescence  532  

4.3  In Vivo Detection  533  

4.3.1  Chemical  533  


4.3.3  Radiological  536  

5.  CBRN FORENSIC ANALYSIS  538  

5.1  Dealing with CBR contaminated evidence  538  

5.2  Forensic study of CBRN materials  538  

5.2.1  Chemical  538  


5.3.3  Radiological and nuclear  541  

5.4  Impact of CBRN agents on standard forensic evidence  541  

5.5  Decontamination of forensic evidence  542  

5.6  Contaminated bodies  543  

6.  CONCLUSION  544  


525

1. Executive Summary 

Terrorist groups remain interested in using CBR materials in attacks and, as a 

result, many countries have been investing in developing a CBRN forensic 

capability. However, the crime scene associated with the preparation or 

release of hazardous chemical, biological, radiological or nuclear (CBRN) 

material provides unique challenges for investigators. Following a terrorist, 

criminal or inadvertent release of CBRN agents, emergency service (first 

responder) agencies may have to deal with mass casualties and public panic, 

where significant numbers of people in the area will seek medical attention 

believing they have been exposed. The risks that these agents pose to first 

responders and crime scene investigators are significant factors in managing 

the response. The CBRN crime scene may be difficult to identify and map, 

particularly following a covert release of agents. The crime scene may be 

both hazardous and fragile, with the potential for critical material to be lost due 

to time delays or lost to the elements, including rain, wind and heat. Efforts to 

decontaminate the scene may also destroy critical forensic evidence. 

Evidence gathered from the scene will be considered contaminated and 

without purpose-build containment laboratories, cannot be examined for 

traditional forensic evidence (fingerprints, fibre, handwriting) until it has been 

decontaminated. The integrity of this evidence may be compromised by the 

agents themselves or by the methods used for decontamination. This article 

is a sequel to the previous review that concentrated on the CBR agents 1 and 

will concentrate on the research carried out since 2007 in support of forensic 

investigation involving CBRN agents. The article is not an exhaustive review 

but will highlight the difficulties CBRN agents create for forensic investigation 

and will give representative examples of the research carried out to overcome 

them. 


In many aspects, a CBRN crime scene is very different to the ones law 

enforcement is used to. Indicators of a CBRN terrorist attack may include 

mass-casualties with symptoms typical of exposure to a CBRN agent and 

claims of responsibility by the perpetrators; however, the covert release of 

CBRN agents, particularly biological and radiological agents, may only be 

recognised as a result of affected people presenting for medical treatment. 

Misdiagnosis and delayed diagnosis, including failure to consider an ‘unusual’ 

cause, such as an act of terrorism, all reduce the likelihood that a terrorist 

event will be detected early and therefore investigated effectively. Critical 

evidence, particularly clinical samples from affected individuals, may have 

been collected and analysed without law enforcement involvement or 

adherence to chain-of-custody principles. This means that locating the site of 

the release and the source of the agent used may require a combined effort 

by law enforcement investigators and public health epidemiologists, who are 

expert at tracing the source of disease outbreaks. Chemical weapons are 

more likely to produce immediate effects, so identifying the crime scene and 

526

the type of weapon used is often more straight-forward. 

Efforts by law enforcement agencies to identify and interdict planning 

processes for CBRN terrorism is a key pillar in preventing CBRN attacks. In 

these cases, law enforcement agencies are required to recognise the 

indicators of a CBRN production/preparation facility (clandestine laboratory), 

as well as other indicators of planning, such as recruitment of scientists from 

universities or legitimate laboratories, trial releases, acquisition of laboratory 

equipment and terrorist training literature/recipes. Crime scene investigators 

need to be aware of the indicators of clandestine laboratories involved in the 

preparation of materials other than illicit drugs. Key indicators include 

dedicated equipment (centrifuges, glassware, detectors), unusual laboratory 

chemicals (agar, culture media, chemical precursors) and possible dispersion 

devices (explosives, gas cylinders, pressurised sprayers). Specific laboratory 

equipment for safe handling of radioactive materials will most likely include 

lead bricks for shielding during storage/preparation and radiation detectors. 

Finally, once evidence has been processed and is not longer needed, specific 

arrangements must be made for the safe destruction and/or disposal of CBRN 

agents and contaminated items. Such items may be considered as hazardous 

materials with specific requirements and reporting. 

The different aspects of investigations involving specific CBRN agents have 

recently been described, 2 as well as the main issues linked with chemical 3 or 

biological 4 agents. However, the forensic investigations related to such events 

may require techniques and protocols not usually available in standard 

forensic laboratories. As a result, many countries are developing some CBRN 

forensic capabilities, usually by agreements with partner agencies that 

specialise in the handling of pathogens or nuclear materials. This review will 

attempt to highlight some of the specific issues arising with CBRN agents and 

give some recent examples of the techniques that are currently being 

developed. 

3. Current CBRN trends 

The ability to stage a successful CBRN attack will depend greatly on the 

technical ability of the terrorist group. While relatively simple “back yard” 

laboratories might allow the preparation of some simple biological and 

chemical agents, more complex laboratories, requiring commercial laboratory 

equipment, would be required to prepare most biological and chemical 

warfare agents, particularly if large-scale or high-purity preparations are the 

desired end-product. 5 There have not been significant CBRN incidents over 

the last 3 years period but the awareness of the disruption that a CBRN attack 

may cause is increasing, hence it is only a question of time before a terrorist 

attack using CBRN materials is performed. 

527

3.1 Chemical 

Chemical warfare agents (CWA) have been used in the past by terrorist 

groups as in the case of the sarin attack in the Tokyo subway in 1995, but 

these high end agents have remained mostly aspirational. The opportunistic 

nature of most terrorist groups has seen a shift towards using more easily 

available toxic industrial chemicals in attacks, such as the recent use of 

chlorine in Iraq to enhance improvised explosive devices. Other toxic 

chemicals of interest are readily generated gases, pesticides or even heavy 

metals such as thallium. 6 

3.2 Biological 

The 2001 Amerithrax case 7 and aspirations of groups like Al Qaida 8 have 

brought anthrax into the limelight and many hoaxes, also known as “white 

powder incidents”, are based on this fear. However, this agent is not that easy 

to prepare and forensic investigators are more likely to encounter other types 

of biological agents. 9 Plague is a potential bioterrorism agent that cannot be 

ignored although several technical hurdles remain. However, many natural 

toxins such as ricin or abrin are readily prepared and can be used to poison 

small numbers of individuals. In general, such toxins are more likely to be 

used than bacteriological agents such as anthrax, or viruses that are even 

harder to culture. 

3.3 Radiological and nuclear 

Although the use of a nuclear device by a terrorist group is highly unlikely, the 

use of radioactive materials to cause harm is a much more likely scenario. It is 

technically infeasible to manufacture radiological material in a clandestine 

laboratory as such preparations require access to a nuclear reactor. 

Nevertheless, the assembly of an Radiological Dispersion Device using 

illicitly-acquired radioactive material and conventional explosives might be 

achieved in a clandestine laboratory. 

4. Detection 

The greatest concern to the investigator is the hazards of the CBRN 

environment, hence rapid identification of the agent is essential. Information 

gathered from the crime scene will help determine the likely impact and risks 

to crime scene examiners and the general public 10 . However, the need for 

personal protective equipment (PPE) can severely limit the capability of crime 

scene investigators to gather appropriate and adequate evidence. If CBRN 

agents have been released in a public place, there will be pressure to rapidly 

528

decontaminate the site to limit the potential for further impact on the 

environment and to restore normal operations. The greatest challenge 

presented to investigators following a covert act of CBRN terrorism may be 

identifying and containing the crime scene. Further, there may be multiple 

crime scenes, including where the agent was manufactured and the site(s) of 

release. 

4.1 Field Detection 

The presence of CBRN agents in a crime scene may not be visible to the 

naked eye, hence a collection approach that maps the crime scene may be 

required. Field detectors will allow the scene examiner to determine the 

location of the agent for sampling. Further, adequate samples will need to be 

collected from within and around the crime scene to determine background 

levels. These data will also be important for site remediation and for declaring 

the previously contaminated site clean. 

A number of field-deployable detectors and screening kits exist that can 

rapidly identify the presence of explosive, radiological, chemical and biological 

agents at a CBRN scene, though the accuracy and sensitivity of these kits for 

specific agents is highly variable. 11 Ultimately, the aim is to be able to 

incorporate the techniques for the detection of CBRN agents, drugs and 

explosives into a single device that could be used by investigators in confined 

places such as clandestine laboratories. 

4.1.2 Chemical 

There are a number of hand held electronic detectors and colourimetric 

indicators that are utilised for presumptive identification of CWA, toxic 

industrial chemicals (TICs) and toxic industrial materials (TIMs). Detection 

principles utilised in equipment available for the detection of chemicals in the 

field include colourimetric, electrochemical, ion mobility spectrometry, flame 

photometry, photoionisation and enzyme based techniques. 12,13 

Field (on-site) and laboratory (off-site) based analytical methods used for the 

detection of chemical warfare agents, their characteristic degradation 

products and related compounds have recently been reviewed, 14 with an 

emphasis on analytical methods developed over the past five to seven years 

which could be used for forensic or other purposes. 

Detectors based on the lab-on-a-chip concept are a field of intense study. 

These detectors, based on microfluidics, could allow fast and cheap field 

detection of a large variety of chemicals including explosive and drugs, which 

could potentially make such a device very useful for clandestine laboratory 

529

investigations. 15,16 Chips based on capillary electrophoresis (CE) are being 

used in the development of novel analysers for chemical and biological threat 

agents. 17 Such a capillary electrophoresis (CE)-based microfluidic device with 

an integrated carbon sensing electrode has been built for the sensing of 

chemicals such as morphine and codeine. 18 

New detectors for chemical warfare agents and their degradation products are 

currently being developed using the biomimetic properties of molecularly 

imprinted polymers (MIPs) that make them attractive for molecular recognition 

and sensing. 19 

Desorption electrospray ionisation-tandem mass spectrometry (DESI-MS) is a 

promising technique for the detection of chemical agents adsorbed on 

surfaces. For example, arsenic species can directly be detected on plant 

tissues without any sample preparation. 20 

Rapid sampling and sample preparation, both in the laboratory and on-site 

can be obtained by using solid-phase microextraction (SPME). Unlike 

traditional sample preparation methods, SPME is a non-exhaustive extraction 

technique in which only a small portion of the target analyte is removed from 

the sample matrix. 21 This method is also commonly used for the study of 

chemical agents (see Forensic study of CBRN agents). 

4.1.3 Biological 

Unlike acts of terrorism involving chemical or radiological agents, the 

investigation of acts of bioterrorism or biocrimes is hindered by the lack of 

real-time detectors for the identification of biological agents. This provides a 

significant gap in the ability of first responders to determine risk when tasked 

with the initial investigation of potential biological releases. 22,23 

To date, immunoassays provide the fastest biological detection technology 

that is commercially available. Most immunoassays use an antibody-mediated 

capture of their target antigens. Typically, detection and identification is by 

means of specific antibodies that are covalently bound with visible or 

fluorescent dyes or with enzymes that generate visible or chemiluminescent 

reaction products. Such a method has been used for the detection of botulism 

neurotoxin A, 24 while a rapid analyte measurement platform (RAMP) as been 

applied to the detection of Bacillus anthracis. 25 

Similar techniques have been used for the detection of ricin. A combination of 

immunocapture of the toxin, examination of the ricin protein on a DNA 

substrate that mimics the toxin's natural RNA target, and analysis of the tryptic 

fragments of the actual toxin, followed by mass spectrometry, allowed the 

530

successful identification of ricin. 26 However, the major downfall of 

immunoassay technologies is that they are currently limited to one or a few 

targets in each test. 

A recent study has evaluated 3 cost-effective biological screening tools 

capable of non-specifically determining the presence of a biological material, 

such as ricin or anthrax, in a suspicious powder. 27 Evaluation of the Guardian 

Bio-Threat alert (BTA) system has shown that it was capable of detecting 

several toxins, such as staphylococcal enterotoxin B (SEB), botulinum toxins 

(BTX) A and B, and ricin with no interference from white-powdered materials 

or coloured matrices; although temperature and toxin sub-types proved 

important factors to take into account. 28,29 

Genetic techniques like the polymerase chain reaction (PCR) and microarrays 

provide the high sensitivity required for the detection of biological agents in 

the field and those samples submitted to the laboratory. To date, real time 

PCR (RT-PCR) technology has shown the greatest application for the genetic 

detection and identification of biological agents. 30 One of the major 

disadvantages of RT-PCR is the inability to multiplex numerous simultaneous 

targets. 

Another technology is based on microarrays, consisting of a bank of nucleic 

acid probes bound to the surface of a glass slide. The advantage of 

microarrays is that the technology allows the simultaneous analyses of 

hundreds to thousands of targets, but traditional microarray technology is 

lengthy, complex, and technically difficult. The integration of automation and 

software processing has largely removed human involvement into many of the 

repetitive tasks involved in the processing and interpretation of microarrays. 

Recent work on the combination of nucleic acid amplification, lateral flow 

chromatography and microarray technologies provides a novel platform from 

which microarrays can be developed for field deployment. 31 

Other methods are also appearing, such as biosensors based on the surface 

plasmon resonance technique, where the surfaces were prepared with 

antibody fragments specific for the biological toxin ricin. 32,33 

Stand-off detection has also attracted some attention as it does not require 

contact with the agent. The detection and discrimination of the biological 

warfare agent surrogate Bacillus subtilis (BG) at a distance of 20 m was 

achieved using standoff laser-induced breakdown spectroscopy (ST-LIBS) 

and linear correlation. 34 

A covert biological attack is likely to be detected by medical practitioners in 

the first instance, but there is a high likelihood that the diseases arising from a 

biological agent may be misdiagnosed due to their rarity. For example, 

531

detection systems for plague may now be needed in non-endemic regions 

and the existing rapid tests for the diagnosis of plague have been reviewed. 35 

4.1.4 Radiological 

The techniques for the detection of alpha, beta, gamma and neutron radiation 

are well established and current research in the field is mainly focussed in 

increasing the sensitivity of these methods. The current state of commercial 

instruments has recently been reviewed for different categories of detection 

equipment including personal radiation detectors, radioisotope identifiers, 

man-portable detection equipment, and radiation portal monitors. In addition, 

emerging technologies are being examined, such as spectroscopic detectors 

and advanced spectroscopic portal monitors. 36 . The principle of operation and 

properties of several types of detectors have also been reviewed, briefly 

detailing the functioning of ionization and wire chambers, semiconductor 

detectors, and scintillation detectors. 37 

4.2 Optically-Stimulated Luminescence 

Optically-Stimulated Luminescence (OSL) is a detection technique that allows 

the detection of items that have been in close proximity with a radioactive 

source. This technique has great potential for counter-terrorism, in helping to 

determine locations where a source or a radioactive device may have been 

stored. 

Ionizing radiation, from a radioactive source, can create charges within 

mineral crystals that may be present around it (concrete, ceramics, even 

mineral pigments in some paints). These charges can become localized, or 

trapped, and held there over significant time scales. Exposure to heat, light, or 

high pressures can release the charges, which is often accompanied by a 

detectable release of light. OSL provides therefore a useful tool for forensic 

investigations and for tracking the former locations of a radioactive source. 38,39 

Recent instrumental developments that can make this method more field 

portable have been reviewed. 40 

A study investigated the OSL response of electronic components found within 

portable electronic devices such as cell phones and pagers, portable 

computers, music and video players, global positioning system receivers, 

cameras, and digital watches. The surface-mount resistors with alumina 

porcelain substrates of such items consistently produced OSL following 

gamma irradiation, allowing to determine whether such an object has been 

close to a source. 41 The same useful property has been found for alumina-rich 

electronic components from a USB flash drive. 42 It has even been shown that 

some epoxy resins used to encapsulate electronic chips produce some OSL 

effects. This is due to the silica used in some cases as filler in the resin. 43 

532

The same technique has been used to determine whether red brick have 

been in contact with beta radiations, 44 and even some glasses have been 

found to be suitable for some dosimetric applications. 45 Gypsum wallboards 

have also been shown to display OSL after irradiations as low as 500 mGy. 46 

4.3 In Vivo Detection 

The covert release of CBRN agents, particularly biological and radiological 

agents, may remain unrecognized for some time. Accurate and fast diagnosis 

will be important to provide fast treatment or to prevent further people being 

affected, but doses or internal physiological effects that could help 

determining the time since exposure will also be essential in order to locate 

the crime scene. It is also anticipated that many terrorist attacks using CBR 

agents may not be very effective or lethal, hence the study of metabolites may 

provide the only indication that people have been exposed. Another direction 

for the research into CBR metabolites is the need for some fast screening 

methods that will allow first responders to separate people needing treatment 

after exposure from the “worried well”. However, at this stage, most 

techniques described below remain laboratory techniques that would be 

difficult to apply in the field. 


The perceived threat of the use of nerve agents by terrorists against civilian 

targets implies the need for methods for point-of-care (POC) diagnosis. The 

methods that are currently available for the assessment of exposure to nerve 

agents are mostly mass-spectrometry based, but new technologies are 

emerging that allow immunochemical detection of acetylcholinesterase that is 

inhibited by nerve agents. These latter techniques could be more suitable for 

field deployment. 47 

Tandem mass spectrometry combined with liquid- or gas-chromatography 

(LC-MS/MS or GC-MS/MS, respectively) have become methods of choice for 

the analysis of toxic chemicals present in the body and the metabolites that 

can be produced after exposure to toxic chemicals. Metabolites, proteins, 

and/or DNA adducts have been identified for the majority of nerve agents and 

vesicants. 48,49 For example, phosphylated butyrylcholinesterase has been 

found to be an important biomarker for the confirmation of exposure to nerve 

agents. This compound can be analysed by LC-MS/MS by the detection of a 

phosphylated nonapeptide produced after the digestion of 

butyrylcholinesterase (BuChE) with pepsin. 50 Several methods, based on 

isotope dilution LC-MS/MS, have been developed for the determination of the 

metabolites of organophosphorus nerve agents such as VX, rVX, Sarin (GB), 

Soman (GD), or Cyclosarin (GF) in human urine. 51,52 

533

Other methods to detect organophosphorus nerve agents exposure consist of 

refluoridation of nerve agents adducted to the butyrylcholinesterase prior to 

solid-phase extraction and analysis by isotope-dilution gas chromatographymass 

spectrometry, 53 or by analysis of the dialkylphosphates metabolites 

found in urine by GC-MS/MS. 54 The fluoride ion-based regeneration method 

has been used successfully in a number of cases. For example, it allowed the 

quantification of VX-butyrylcholinesterase adduct in plasma, resulting from a 

relatively low-level accidental human exposure case, by gas chromatographyhigh 

resolution mass spectrometry of the resulting biomarker ethyl 

methylphosphonofluoridate (VX-G), 55 or by GC-MS/MS of VX-G in red blood 

cells. 56 The same method was applied to the detection of VX-G in Gottingen 

minipig red blood cells after whole-body exposure to VX vapour, 57 while VX in 

plasma could be identified directly. 58 

The fluoride method was also applied to analysis of soman in rat plasma. 59 

The relative toxicity of the four soman stereoisomers was also investigated. 60 

A new method, based on high-performance liquid chromatography-mass 

spectrometry with negative ion electrospray ionisation and time-of-flight 

detection (LC-ESI-MS-TOF), has been developed and validated for the 

determination of isopropyl methylphosphonic acid (IMPA) and cyclohexyl 

methylphosphonic acid (CMPA), the metabolic hydrolysis products of sarin, 

and cyclosarin, respectively. 61 

A lab-on-a-chip method has been developed for the detection of sarin in 

blood, based on continuous-flow microfluidics with sequential stages for lysis 

of whole blood, regeneration of free nerve agent from its complex with blood 

cholinesterase, protein precipitation, filtration, enzyme-assisted reaction, and 

optical detection. 62 

Exposures to vesicants such as sulphur mustard (HD) or nitrogen mustard 

(HN) can also be determined by metabolites analysis. For example, exposure 

to HD can be ascertained by measuring thiodiglycol (TDG), TDG-sulphoxide, 

and the bis-mercapturate of mustard sulphone in urine. 63 Other metabolites 

such as adducts of HD with albumin, with glutamic and aspartic acids of 

plasma proteins or with haemoglobin have also been extensively 

studied, 64,65,66 while detection of blood protein adducts can be identified from 

the cleavage of protein-hydroxyethylthioethyl esters to produce thiodiglycol. 67 

The albumin-derived adduct biomarkers present in blood samples have also 

been investigated for nitrogen mustards (HN-1, HN-2, and HN-3). 68 

The determination of the sesqui-mustard metabolites bis(2hydroxyethylthio)alkanes 

(n = 1-5) and the oxy-mustard metabolite bis(2hydroxyethylthioethyl)ether 

in human urine by HPLC-MS/MS could similarly 

provide indication of exposure to sesqui- and oxy-mustards that are potential 

chemical weapons because of their vesicant properties. 69 

534

Biomarkers or new analytical methods for exposures to industrial toxics have 

also been studied. Chlorine exposure has been found to trigger both 3chlorotyrosine 

(CY) and 3,5-dichlorotyrosine (dCY) – produced by the 

modification of tyrosine by reactive chlorine – making them promising new 

metabolites to investigate. 70 Using an LC-MS/MS method, six rodenticides 

could be detected and quantified after liquid/liquid extraction of blood 

samples. 71 

A simple method, based on liquid chromatography-mass spectrometry (LC- 

MS), has been developed and validated for the determination of fenitrothion, 

its metabolite 3-methyl-4-nitrophenol, and five other organophosphorus 

pesticides (acephate, cyanophos, methidathion, ethyl-p-nitrophenyl 

phenylphosphonothioate, and malathion) in human urine. 72 

A gas chromatography-mass spectroscopic (GC-MS) method has also been 

described for the routine assessment of organo-phosphorus pesticide 

exposure via automated solid phase extraction of the metabolites present in 

urine prior to derivatisation with pentafluorobenzyl bromide and quantitation 

by GC-MS, 73 while the same method was used for the determination of 

dichlorvos in blood samples. 74 

Inorganic materials in body samples can also be analysed successfully. A 

rapid and reliable method, based on high-performance liquid chromatography 

(HPLC)-inductively coupled plasma-mass spectrometry (ICP-MS), has been 

developed for the determination of inorganic arsenic in human urine. 75 The 

same method was used to analyse Lewisite metabolites in urine. 76 

A headspace gas chromatographic method with electron capture detection 

has been applied to the determination of cyanide in blood, 77 while highperformance 

liquid chromatography with inductively coupled plasma mass 

spectrometry (HPLC-ICP-MS) was used to analyse arsine-exposed human 

blood samples for arsenic speciation, 78 or arsenic in urine. 79 

A spectrophotometric method has been developed for the analysis of 

inorganic arsenic in urine but the samples need to be treated with copper 

granules prior to reaction with ammonium molybdate for visual detection. 80 

Inductively coupled plasma optical emission spectrometry (ICP-OES) has 

been found suitable for the determination of mercury and lead in biological 

fluids and tissues at concentrations likely to be encountered in cases of acute 

poisoning, but the sensitivity remains weak for lower exposures. 81 

Toxic gas such as phosphine, present in body samples, could be detected by 

using headspace gas chromatography with nitrogen-phosphorus detection 

(HS-GC/NPD), 82 while concentrations of thiosulphate in urine could indicate 

exposure to hydrogen sulphide gas. 83 

535


Intensive work has been carried out to determine levels of toxins in the 

body, 84 several using new liquid chromatographic-tandem mass spectrometric 

(LC-MS-MS) methods. Such a method has been developed for the 

simultaneous determination of aconitine, mesaconitine, hypaconitine, and 

jesaconitine in whole blood, 85 aconitine in blood and urine, 86 or has been used 

for the identification of cytosine in post-mortem samples. 87 Saxitoxin and 

neosaxitoxin in urine samples preconcentrated by solid-phase extraction 

could be identified by electrospray ionisation and tandem mass 

spectrometry. 88 Multi-analyte procedures have been developed for the 

detection and validated quantification of the toxic alkaloids aconitine, atropine, 

colchicine, coniine, cytisine, nicotine and cotinine, physostigmine, and 

scopolamine in plasma using liquid chromatography-atmospheric pressure 

chemical ionisation-mass spectrometry (LC-APCI-MS) and liquid 

chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI- 

MS/MS) after solid-phase extraction. 89 

Ricinine, a piperidine alkaloidal toxin found in the castor bean, is often 

described as a biomarker for exposure to ricin. This biomarker was recently 

successfully identified in blood, urine, and vitreous humour specimens using 

solid-phase extraction and liquid chromatography-tandem mass spectrometry 

(LC-MS/MS) with an electrospray source in positive ionisation mode. 90 

Abrin – a toxic protein similar to ricin that is found in the jequirity (or rosary) 

seed – has been analysed in urine samples subjected to solid-phase 

extraction prior to high-performance liquid chromatographic (HPLC) 

separation and mass spectrometric analysis. 91 

A different method to identify exposure to ricin is based on a sensitive 

luminescent assay detecting the adenine produced by a ribosome 

depurination catalysed by ricin. 92 

More sophisticated methods are also starting to appear, such as the use of 

nano-electrospray hybrid quadrupole time-of-flight mass spectrometry for the 

accurate identification of anatoxin A. 93 

4.3.3 Radiological 

Medical intervention in the aftermath of a radiological or nuclear incident will 

require rapid assessment of the extent of exposure of those involved, usually 

via the determination of the internal dose received. 94 A recent study has 

compared different methods of inductively coupled plasma mass spectrometry 

(ICP-MS) for the determination of plutonium in urine with a thermal ionisation 

536

mass spectrometry (TIMS) method and two alpha spectrometry methods. 95 

However, the main field of research is concerned with biodosimetry. 96 

Biodosimetry reflects the radiation dose absorbed by an individual. It is not a 

direct, real-time reading of dose, but is based on clinical observations and 

laboratory results used to estimate the radiation dose received after exposure 

or internal contamination. 97 Extensive work is being carried out in this field 

with a large variety of techniques, including the development of reliable and 

simple mathematical and statistical tools to analyse the observed data. 98 

There are several methods that can be used in biodosimetry that will only be 

briefly mentioned in this paper as each of them would require a review of their 

own. 99 Measurements based on physical methods use dose estimation of 

physical changes in samples such as teeth and nails with techniques such as 

EPR or spectroscopy. More recently, optically stimulated luminescence (OSL) 

properties of dental enamel have been investigated as a tool to determine 

exposure to radiation. The potential and challenges of OSL for filling the 

technological gap in biodosimetry required for medical triage following a 

radiological or nuclear accident or terrorist event have been reviewed. 100 The 

dose response and thermal stability characteristics of an adult human molar 

tooth to blue photon stimulation have been reported. Statistically significant 

dose response OSL was observed for beta doses ≥ 1.4 Gy. 101 

Biological methods are based on the measure of biological processes that are 

affected by radiations, such as abnormal cell divisions or on clinical 

parameters such as lymphosite count, chromosomal aberrations, the 

identification of specific biomarkers resulting from irradiated organs or timing 

of nausea or vomiting. Cytogenetic dosimetry, based on chromosomal 

aberrations, is probably the most widespread method and one aspect of it, 

chromosome translocations due to radiations, has recently been reviewed. 102 

However, while the traditional radiation exposure biomarkers based on 

cytogenetic assays serve as standards, the development of rapid and noninvasive 

tests for radiation exposure is needed. Genomics based knowledge 

is providing new avenues for investigation. The examination of gene 

expression after ionizing radiation exposure could provide potential molecular 

markers for biodosimetry. 103 Some promising macromolecules found in 

normal (without tumour response) irradiated tissues have been identified. 104 

Irradiated human cells have recently been screened, and 55 ionizing-radiation 

responsive proteins have been identified. 105 

537

5. CBRN forensic analysis 

5.1 Dealing with CBR contaminated evidence 

The rarity and nature of CBRN criminal and terrorism events means that 

investigations tend to be extensive and protracted. This may necessitate longterm 

storage of the agents and contaminated items, including biological 

specimens, computers and vehicles. Many CBRN agents, or the solvents in 

which they are collected, can react with standard evidence storage containers 

(notably plastic bottles or bags), potentially resulting in the loss of evidence 

and harm to people and the environment. In addition, some plastics in storage 

bags and tubes can absorb chemical and biological agents, thus reducing the 

yield of agent for analysis or altering its properties. As most CBRN agents are 

unstable, they will degrade over time, losing their viability or potency and even 

changing their chemical properties. In some cases, it is possible to treat the 

agents to minimise loss (such as freezing, drying, or immobilizing in a matrix), 

but all of these treatments will alter the properties of the agent from its original 

condition. 

Most standard forensic laboratories and law enforcement agencies do not 

have appropriate facilities for safe, long-term storage of CBRN agents and 

contaminated items. 106 These items may need to be stored in facilities 

dedicated to the analysis or research of specific material, where capacity to 

store large or multiple items may be lacking. Consideration must be given to 

maintaining security, evidence integrity and chain-of-custody for storage of 

CBRN items in facilities that are not managed by law enforcement agencies. 

5.2 Forensic study of CBRN materials 

Specific disciplines are required for the direct examination of CBRN agents 

and their production/preparation processes, providing information that can be 

used for the attribution of CBRN crime. As with any forensic field, the 

application and interpretation of data is critical in excluding a particular source 

as the origin of the sample, as well as attributing the sample to a particular 

source. This is particularly important in discriminating malicious use of CBRN 

agents from natural causes. 


The analysis of chemical reagents, chemical degradation products and 

equipment found at the scene can determine the types agents being 

manufactured and the preparatory method. Chemical trace analyses, 

including isotopic ratios and mass spectrometry, are not only useful for 

chemical tracing but can support microbial forensics to determine the origin of 

538

source material (such as toxic plants) and the origins of growth media. 

A new analytical protocol for the forensic identification of chemical warfare 

agent surrogates for sulphur mustard, sarin, VX, soman and tabun, based on 

rapid solid-phase microextraction followed by gas chromatographic-mass 

spectrometric (GC-MS) analysis, has been developed for various media. 107 

Desorption electrospray ionization mass spectrometry (DESI-MS) is a 

promising new technique for forensic applications. It has been applied to the 

direct analysis of CWA samples collected on swabs or solid-phase 

microextraction fibres without the need for additional sample handling. 108 

A simple and rapid generic method has also been developed for the 

simultaneous extraction of various classes of pesticides, mycotoxins, plant 

toxins, and veterinary drugs in a range of matrices for subsequent analysis by 

liquid chromatography-mass spectrometry (LC-MS). The new generic 

extraction method was validated for 136 pesticides, 36 natural toxins, and 86 

veterinary drugs in compound feed and honey using ultraperformance liquid 

chromatography-tandem mass spectrometry (UPLC-MS/MS) for analysis. 109 

One- and two-dimensional nuclear magnetic resonance (NMR) spectroscopy 

can also be applied to the identifications of unknown powders. 110 

The analysis of impurities in warfare agents to provide forensic signatures is 

field that has attracted more attention in recent time. The use of bioinformatic 

and chemometric tools on liquid chromatography-mass spectrometry (LC-MS) 

data has proved very useful for the discovery of trace forensic signatures for 

sample matching of ten stocks of the nerve-agent precursor known as 

methylphosphonic dichloride (dichlor). 111 The feasibility of using analytical and 

chemometric methodologies to reveal and exploit the chemical impurity 

profiles from commercial di-Me methylphosphonate (DMMP) samples has 

been explored to illustrate the type of forensic information that may be 

obtained from chemical-attack evidence. 112 Alkyl methylphosphonic acids 

(RMPAs) and methylphosphonic acid (MPA), both nerve gas hydrolysis 

products, can be readily identified by GC-MS after having undergone tertbutyldimethylsilylation. 

113 


Since the Amerithrax events of 2001, a new scientific discipline has appeared 

– “microbial forensics” – that is dedicated to the analysis of evidence from an 

actual or threatened bioterrorism act, biocrime, or inadvertent 

microorganism/toxin release for attribution purposes. 114 The forensic 

examination of microbial agents will be most effective if there is sufficient 

basic scientific information concerning microbial genetics, evolution, 

physiology and ecology of the agent. 115,116 

539

Apart from providing greater confidence in the data for attribution purposes, 

understanding the ‘microbial background’ of relevant geographic areas is 

critical for determining the source of the biological agent for consequence 

management and for remediation. For example, a catchment-scale microbial 

source tracking study in the Leven Estuary drainage basin in northwestern 

England has been carried out. Although discrimination between livestock- and 

human-derived faecal indicator concentrations could be obtained, the study 

failed to provide quantitative source apportionment for the catchment 

studied. 117 

Mass spectrometry (MS) has proved a useful tool for microbial forensics, 

allowing the identification of microorganisms through their microbial 

biomarkers rapidly and reliably without time-consuming sample pre-treatment. 

These MS techniques have recently been reviewed. 118 Three different mass 

spectral techniques were used to characterise the culture medium of Bacillus 

spores to predict the production environment of unknown-source 

microorganisms. The data of the three techniques were integrated via a 

Bayesian statistical framework, significantly improving the accuracy and 

robustness of the analyses. 119 

Electrospray ionisation tandem mass spectrometry (ESI-MS) is a promising 

emerging technique for microbial forensics. This technique was used for the 

detection of trace agar residue within suspect microbial samples, indicating 

that the potential agent was produced using agar-solidified growth media. 120 

This method, coupled with PCR (PCR/ESI-MS), enables rapid detection, 

identification, and serotyping of pathogens. The assay employs a highperformance 

mass spectrometer to “weigh” the amplicons obtained from PCR. 

The measured masses are then converted to base compositions and, by 

comparison against a database of the genetic sequences, the serotype 

present in a sample can be determined. 121,122 

Hyperspectral imaging-enabled microscopy has also been evaluated for the 

identification of chemical signatures in simulated bioagent materials. 123 

The sampling of the biological agent is also important. The optimal 

procedures for testing everyday surfaces likely to be contaminated following 

the release of environmentally stable biological weapons such as anthrax 

spores and ricin have been studied, showing that dry devices, including 

swabs, wipes, and Trace Evidence Collection Filters, had poor retrieval 

efficiencies. 124 

In order to get a forensic signature for ricin extracts to enable cultivar and 

provenance determination, seeds from 14 Ricinus communis specimens were 

collected from the east coast of Australia and their metabolome – the 

540

complete set of small-molecule metabolites to be found within a biological 

sample – analysed by high pressure liquid chromatography with UV detection 

(HPLC-UV), liquid chromatography–mass spectrometry (LC-MS), and 1 H 

NMR spectroscopy. 125 

5.3.3 Radiological and nuclear 

“Nuclear forensics” is the branch of science that seeks to examine the nature, 

use and origin of intercepted illicit nuclear or radioactive materials. 126,127 The 

analysis seeks to produce a characteristic energetic “signature” for the 

material as evidence for attribution. It can be applied to investigations 

involving the malevolent use of nuclear materials and is instrumental in 

determining adherence to international safeguards for illicit trafficking. 

Nuclear forensic analyses are also essential to determine the origin of the 

radioactive material. 

New methods for the detection of previous neutron irradiation and 

reprocessing of illicit nuclear materials have been reported for the 

characterisation of nuclear materials of unknown origin in nuclear forensics. 

Alpha spectrometry and inductively coupled plasma sector-field mass 

spectrometry (ICP-SFMS) were used to measure traces of uranium and 

plutonium isotopes derived from previous neutron irradiation of uraniumcontaining 

nuclear materials. 128 A similar laser ablation assisted inductively 

coupled plasma-mass spectrometry (ICP-MS) was used for the rapid 

classification of nuclear forensic evidence based on uranium isotopic 

composition of seized samples. 129 

Confirmation of the origin of a plutonium sample found in Hanford USA in 

2004 as being the X-10 reactor in Oak Ridge, Tennessee, was obtained from 

the analytical results of minor actinide isotopes and reactor model 

simulations. 130 

5.4 Impact of CBRN agents on standard forensic evidence 

The CBRN agents themselves, particularly radiological and chemical agents, 

can damage or destroy critical evidence, including biological evidence (DNA, 

hair, fingerprints), as well as documents, computer data and other reactive 

material. Standard forensic analysis for DNA, fingerprints, handwriting from 

evidence gathered from a CBRN crime scene will be unsafe if evidence is not 

handled with physical protection or decontaminated. 

It has been shown that DNA can be safely recovered from blood samples 

having been exposed to chemical agents – sulphur mustard, sodium 

fluoroacetate, sarin and diazinon – after extraction of the sample, 131 although 

541

fresh and aged bloodstains exposed to hydrogen cyanide, phosgene, 

chlorine, dimethyl sulphate, sodium fluoroacetate, diazinon, sulphur mustard, 

lewisite I and sarin, in gaseous and/or liquid form tended to give lower 

concentrations of DNA. 132 

The enhancement of latent fingerprints with ninhydrin or cyanoacrylate has 

been found to be inhibited by the exposure of such evidence to either 

hydrogen chloride or chlorine, resulting in acidification. In such cases, pretreating 

the samples with volatile bases, such as triethylamine or 

ethanolamine, is required to obtain a successful visualisation of the latent 

fingerprints. 133 

The effect of aerosolised bacteria on fingerprint impression evidence was 

studied by contaminating latent and blood-contaminated fingerprints 

deposited on various surfaces with bacteria which serve as standard 

simulants for anthrax, plague, and tularaemia. It was found that all of the 

fingerprint reagents tested performed well in the presence of these biological 

agents. 134 

Irradiation of forensic evidences may arise from the release of a radioactive 

compound but can also be the result of decontamination of evidences 

contaminated with biological agents by irradiation (see also Decontamination 

of forensic evidence). 

Radiations can have significant impact on fingerprints. Latent fingerprints from 

surfaces such as aluminium, glass, paper, and plastic, were exposed to doses 

of radiation ranging from 1 to 1000 kGy. With the exception of glass and 

aluminium substrates, radiolysis was found to have a considerable effect on 

the quality of the developed prints. 135 

5.5 Decontamination of forensic evidence 

Most forensic laboratories do not have facilities that allow physical 

examination of contaminated evidence, so decontamination is often the only 

option. Chemical, heat and irradiation methods used for decontamination 

have the potential to significantly alter surface materials as well as affecting 

the integrity of latent fingerprints, DNA and the characteristics of writing inks 

and dyes. 

A study of bloodstains exposed to chemical warfare agents and 

decontamination agents were subjected to DNA extraction and typing. DNA 

profiles were not obtained from blood dilution samples exposed to the 

CASCAD and MDF LSA-100 decontamination products. 132 

542

One technique to decontaminate evidences contaminated with anthrax spores 

is to expose the evidence to formaldehyde gas. A new protocol for such 

decontamination was developed that allows the destruction of viable spores 

without affecting the successful recovery of latent fingerprints. 136 

The effect of electron beam irradiation – used for disinfecting letters 

contaminated with biological agents – on various writing inks was 

investigated, showing only little degradation of the inks. 137 A similar study 

sought to determine whether gamma irradiation could be used as an effective 

biological decontaminant. Its effect on the recovery of latent fingermarks and 

DNA was tested and established that gamma irradiation was a viable 

decontamination option, 138 although an earlier study indicated that the 

irradiation process can have a detrimental effect on the success of certain 

visualization reagents. 139 

The efficacy of two decontamination techniques (chemical and physical) 

designed for the removal of radiological material dispersed on documents 

have been assessed. The impact that these procedures have on the 

development of latent fingermarks and the forensic analysis of the inks on 

these documents has also been studied. It was found that slight changes in 

the colour and chemical composition of a variety of document inks and a 

destruction of fingermark ridges occurred after chemical decontamination, but 

physical decontamination had no impact on these parameters. 140 

5.6 Contaminated bodies 

Bodies contaminated with CBRN agents provide their own challenges. The 

bodies themselves may require decontamination, but internal organs may still 

retain some of the toxic agent. A guide for medical examiners and coroners 

has recently been issued, providing information and suggestions about the 

principles involved in decontamination procedures as they may impact on 

evidence collection and/or the temporal sequence of death investigation. 141 

The risks associated with contaminated CBR fatalities depend on the nature 

of the hazards and a number of environmental factors encountered during 

handling. The UK Government has developed body process pathway 

algorithms to provide guidance to those involved at all stages of contaminated 

mass fatalities. 142 The situation is even more complicated for radioactively 

contaminated fatalities, where regulations about the transport and storage of 

radioactive material may apply. 143 

The requirement for identification of the deceased may necessitate the 

removal of contaminated samples to a DNA laboratory for processing. The 

potential risks and difficulties associated with processing samples of this type 

have been reviewed. 144 

543

6. Conclusion 

The nature of a CBRN crime scene is quite different to other ones. It is an 

aggressive environment that requires the investigator to be protected and that 

will make the collection of evidence difficult. Detection techniques for chemical 

and radiological agents are adequate but a method for rapid identification of 

biological agents is still lacking. Detection of the exposure to CBR agents has 

come a long way, with biomarkers being identified and the field of 

biodosimetry being developed. However, the handling of the evidence 

remains a problem. Techniques to investigate the agents, especially regarding 

their source attribution, are being developed. Meanwhile obtaining good 

forensic evidence from contaminated items remains difficult although more 

harmless decontamination methods are being investigated. 

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188. 

555

143 Wood CM, DePaolo F, Whitaker RD. Guidelines for Handling Decedents 

Contaminated with Radioactive Materials. Report from the US Department 

of Health and Human Services and the Centers for Disease Control and 

Prevention 2007. 

144 Socratous E, Graham EAM. DNA reviews: DNA identification following 

CBRN incidents. Forensic Science, Medicine, and Pathology 2008; 4(4): 

255-258. 

556

Environmental crime 

Review : 2007 - 2010 

Jennifer A. Suggs, M.S. 

EPA NEIC 

P.O. Box 25227 

Building 25, Denver Federal Center 

Denver, Colorado USA 80225-5227 

suggs.jennifer@epa.gov 

557


1.  Introduction  561  

2.  Environmental crime  562  

3.  Environmental forensics  566  

3.1  10 years of environmental forensics  566  

Table 1.  Topics in Selected Environmental Forensics Review Papers  567  

Table 2.  Survey of Recent Topics Published in Environmental Forensics (2007–May 2010)  570  

3.2  recent books and papers  571  

Table 3.  Recent Papers in the Field of Environmental Forensics  572  

4.  Electronic waste  577  

5.  Regulatory and legislative considerations  578  

6.  Contamination and health effects  580  

7.  Analytical methods  582  

Table 4.  Publications from the Townsend Research Group, University of Florida, Gainesville  584  

8.  Biological waste  585  

8.1  Concentrated animal feeding operations (cafos)  585  

8.1.1  Environmental Impacts  585  

8.1.2  Analytical Methods for Emissions  587  

8.1.3  Methods for Tracking Waste Releases  588  

8.2  Microbial source tracking  589  

9.  Acknowledgments  592  

10.  References  592  

558

LIST OF ACRONYMS 

AFO – animal feeding operation 

ARF – advance recovery fee 

ASTER – Advanced Spaceborne Thermal Emission 

and Reflection Radiometer 

BAN – Basel Action Network 

BTEX – benzene, toluene, ethylbenzene, and 

xylene 

CAA – Clean Air Act 

CAFO – concentrated animal feeding operation 

CERCLA – Comprehensive Environmental 

Response, Compensation, and Liability Act 

CF-IRMS – continuous flow-isotope ratio mass 

spectrometry 

CPU – central processing unit 

CRT – cathode ray tube 

CWA – Clean Water Act 

DART – direct analysis in real time 

DIN S4 – extraction method of the Institut fur 

Normung, Germany 

DNA – deoxyribonucleic acid 

DTSC – California Department of Toxic Substance 

Control 

EA NEN 7371/NEN 7371 – Dutch Environmental 

Agency Availability Test 

ECD – electron capture detector 

EDXRF – energy-dispersive X-ray fluorescence 

EEE – electrical and electronic equipment 

EIA – Environmental Investigation Agency 

EPR – extended producer responsibility 

ERI – electrical resistivity imaging 

ESA – Endangered Species Act 

EU – European Union 

e-waste – electronic waste 

MS – mass spectrometry 

MST – microbial source tracking 

MSW – municipal solid waste 

MTBE – methyl tert-butyl ether 

NAPL – non-aqueous phase liquids 

NCI – negative chemical ionization 

NMP – Nutrient Management Plan 

NPDES – National Pollutant Discharge Elimination 

System 

NZEC – New Zealand Environmental Court 

OCP – organochlorine pesticide 

OP-FTIR – open path Fourier transform infrared 

spectroscopy 

PAH – polycyclic aromatic hydrocarbon 

PBDD – polybrominated dibenzo-p-dioxin 

PBDF – polybrominated dibenzofuran 

PCB – polychlorinated biphenyl 

PCE – tetrachloroethene 

PCR – polymerase chain reaction 

PDA – portable digital assistant 

FID – flame ionization detector 

FIFRA – Federal Insecticide, Fungicide, and 

Rodenticide Act 

FTIR – Fourier transform infrared spectroscopy 

GC – gas chromatography 

GC-ECD – gas chromatography-electron capture 

detection 

GC-FID – gas chromatography-flame ionization 

detection 

GC/HRMS – gas chromatography/high-resolution 

mass spectrometry 

GC-IRMS – gas chromatography-isotope ratio 


GC-MS, GC/MS – gas chromatography-mass 

spectrometry 

GC-NCI-MS – gas chromatography-negative 

chemical ionization mass spectrometry 

GIS – geographic information systems 

GPR – ground-penetrating radar 

GPS – global positioning system 

HARL – Home Appliance Recycling Law (Japan) 

HF-LPME – hollow fiber based liquid-phase 

microextraction 

ICPMS – inductively coupled plasma-mass 

spectrometry 

IRMS – isotope ratio mass spectrometry 

LA – laser ablation 

LA-ICPMS – laser ablation inductively coupled 


LPEUR – Law for Promotion of Effective Utilization 

of Resources (Japan) 

LPME – liquid-phase microextraction 

MODIS – moderate resolution imaging 

spectroradiometer 

SRM – standard reference material 

SW-846 – US EPA Test Methods for Evaluating 

Solid Waste, Physical/Chemical Methods 

TCA – trichloroethane 

TCE – trichloroethene 

TCLP – Toxicity Characteristic Leaching Procedure 

TSCA – Toxic Substances Control Act 

TTLC – Total Threshold Limit Concentrations 

TV – television 

UEEE – used electrical and electronic equipment 

US – United States 

US EPA – United States Environmental Protection 

Agency 

UV – ultraviolet 

VTB – viral toolbox 

VOC – volatile organic compound 

WEEE – waste electrical and electronic equipment 

WET – Waste Extraction Test 

XRD – X-ray diffraction 

559

LIST OF ACRONYMS 

PDF – portable document format 

PFPD – pulsed flame photometric detectors 

RCRA – Resource Conservation and Recovery Act 

REACH – Registration, Evaluation, and 

Authorization of Chemicals 

RHA – Rivers and Harbors Act 

RoHS – Restriction on Hazardous Substances (EU) 

RT – reverse transcription 

SDWA – Safe Drinking Water Act 

SEM – scanning electron microscope 

SPLP – Synthetic Precipitation Leaching Procedure 

SPE – solid phase extraction 

560


This environmental crime review paper is a follow up to the review prepared for the 15 th 

Interpol International Forensic Science Symposium in October 2007. It is compiled 

primarily from a literature review of papers published since early 2007 on specific 

environmental crime and environmental forensics topics. Books, critical reviews, and 

other papers that were published before 2007 are also included when those may be of 

interest to the reader. 

This paper differs in format from previous environmental crime review papers in that it 

does not present a field or crime scene examination component nor does it summarize 

the wide range of analytical instrumentation and methods useful to environmental 

analysis. For readers desiring some background knowledge for field investigations and 

sample analysis, the review papers prepared for the three prior Interpol International 

Forensic Science Symposia are available. 1-3 

Electronic waste (e-waste) and microbial source tracking are two topics highlighted in 

this review. Concerns over the hazardous components in electrical and electronic 

equipment (EEE) have led to regulations that seek changes in the components of the 

equipment and establish requirements for disposing of that equipment. 4-10 Initiatives for 

reuse, recycling, and proper disposal of e-waste are underway in many nations, but the 

success of the initiatives and the compliance with regulations vary. 11-15 Literature 

searches reveal many papers on the contamination and health effects from improper 

recycling of waste EEE (WEEE). 16-20 Many papers discuss the leachability of metal 

contaminants from e-waste with reference to the United States Environmental 

21, 22 

Protection Agency (US EPA) Toxicity Characteristic Leaching Procedure (TCLP), 

23, 24 

although metals are not the only contaminants of concern. 

Microbial source tracking (MST) involves the various biological and chemical techniques 

that can be used to determine a source of fecal pollution. Both human and animal 

waste can contaminate water sources, and the origins and causes for contamination 

events must be determined in order to prevent repeat occurrences. 25, 26 Several groups 

are investigating methods that may help locate a single source or multiple contributing 

sources of a contamination. 27-30 

Environmental crime and environmental forensics are large, general search topics. The 

environmental crime search yielded papers with specific regulatory and enforcement 

reviews 31-34 and legal perspectives on environmental crime cases. 35-37 

An essential environmental forensics journal turns 10 years old; some of its history and 

fundamental papers are revisited. 38 Chemical characterization and determining sources 

of pollutants are key topics in environmental forensics papers. 39-44 A new area of 

environmental forensics investigations was proposed in response to problems with 

imported Chinese drywall. 45 

561

2. Environmental crime 

For the past 5 years, Solow has reviewed United States (US) environmental crime 

enforcement actions from the preceding year. 33, 46-49 For the survey of enforcement 

actions of 2009, Solow and Carpenter summarized notable criminal cases and 

graphically presented the number of cases prosecuted under environmental statutes. 

Also included were summaries of the current US EPA National Enforcement Initiatives, 

the new US Department of Justice guidance on discovery, the increased oversight of 

chemicals in the Toxic Substances Control Act, and the emerging environmental issue 

of e-waste. 33 In addition to the usual summaries of selected cases, Solow’s survey of 

2008 developments included a note about criminal cases on illegal discharges from 

vessels and the decisions on the corporate criminal liability for acts of non-managerial 

employees involved in these illegal discharges. 46 The 2007 survey included an 

interview with Stacey Mitchell, chief of the US Department of Justice Environmental 

Crimes Section, a mention of a highly-rated environmental crimes blog, and discussions 

on Clean Water Act (CWA) prosecutions, voluntary disclosures, and attorney-client 

privilege. 47 

Sanders writes an annual environmental law review for Water Environment Research. 

Each review includes regulatory changes and developments in wastewater treatment, 

ambient waters, hazardous wastes, and emerging issues. Also included are summaries 

of agreements, enforcement actions, and lawsuits from the previous year. 50-52 

Natural Resources & Environment, a journal published quarterly by the Section of 

Environment, Energy, and Resources of the American Bar Association, dedicated an 

issue to environmental crime (Winter 2009). 53 The issue included an interview with 

Granta Nakayama who was the Assistant Administrator for the US EPA Office of 

Enforcement and Compliance Assurance at that time. 54 Harrell et al. discussed the 

“knowing violations” aspect of criminal prosecutions, the variation in penalties among 

the US environmental statutes, and the complexities of prosecuting an environmental 

crime case. The authors explained that an environmental prosecutor would need to 

develop an understanding of statutes, programs, and the multiple layers of regulations 

at federal, state, and local levels and develop the skill to present the complex material to 

a jury. 37 Periconi provided a history of environmental crime prosecutions in the State of 

New York and added information on current trends and case law developments. The 

author described an overall decline in environmental crime prosecutions in New York 

state and suggested that one of the reasons given for this decline in prosecutions was 

the lack of jail time imposed for environmental crimes. Periconi noted that judges and 

juries seem more inclined to impose jail time for more threatening crimes (for example, 

street crime); civil enforcement, with its large fines and monitoring programs for 

environmental compliance, seems to have moved to the forefront of dealing with 

environmental violations. 32 Other topics from the environmental crime articles in the 

issue included the role of motive in environmental crimes, victims’ rights, the 

prosecution of corporate officers, and new European Union (EU) chemical regulations. 53 

562

Burns et al. provided an overview of issues, history, and developments in the wide field 

of environmental crime in their book, Environmental Law, Crime, and Justice. The 

authors briefly reviewed types of environmental harm and detailed the US history of 

environmentalism and how it stimulated actions and policies of the federal government. 

The founding and history of the US Environmental Protection Agency, the development 

and enforcement of environmental laws (including crime investigation and data 

sources), and the connection between environmental justice and enforcement are 

detailed. The authors also shared their perspectives on the future challenges and 

expectations regarding regulatory efforts and criminal prosecutions of environmental 

crimes. 55 

Shafer et al. authored a concise summary of environmental crime as defined by US 

statutes. The article starts with general introductions to criminal versus civil penalties, 

federal enforcement responsibilities, and links with other criminal violations. After a 

brief summary of the “knowing” element of criminal violations, the authors discuss the 

topics of liability, defenses, voluntary compliance, and sentencing. In the following 

sections, the authors examined the Clean Air Act (CAA), the Clean Water Act (CWA), 

the Rivers and Harbors Act (RHA), the Safe Drinking Water Act (SDWA), the 

Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA), 

the Resource Conservation and Recovery Act (RCRA), the Toxic Substances Control 

Act (TSCA), the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA), and the 

Endangered Species Act (ESA). For each, the authors provided the purpose of the 

statute and the elements of an offense of that statute including violations, intent, and 

exceptions. Possible defenses and penalties for violation of the statute are also 

included. While each section in this review is brief, the authors provided more than 600 

footnotes. 36 

Brickey wrote Environmental Crime: Law, Policy, Prosecution, a law school textbook 

dedicated to the study of environmental crime. The book, published in 2009, includes 

discussions on the environmental regulatory framework and the integration of criminal 

law principles with environmental law. Brickey focused on the criminal provisions for 

CWA, CAA, RCRA, and CERCLA and focused on the concept of knowing 

endangerment. Conventional criminal statues often linked to environmental crime 

cases, such as conspiracy, mail fraud, false statements, and obstruction of justice, were 

discussed along with some enforcement issues. Also included were several excerpts 

from legal work by other authors. 56 

Sachs compared the chemical toxicity regulations of the US to those of the EU and 

found the US regulations lacking. The author stated that the Toxic Substances Control 

Act (TSCA) in the US has been hindered by the heavy information demand it places on 

the US EPA. Thousands of chemicals in production at the time TSCA was written were 

exempted from the Act, and, since then, the burden has been on the US EPA to prove 

that a chemical poses an unreasonable risk before it can compel testing by the 

manufacturer. In contrast to this, other US chemical regulations are more strict; 

manufacturers of pesticides and pharmaceuticals are required to prove the safety of 

their chemical products to regulators. Sachs described the EU Registration, Evaluation, 

563

and Authorization of Chemicals (REACH) legislation as the law that will buttress TSCA. 

REACH requires that chemicals manufactured or imported into the EU be registered 

and undergo an amount of toxicity testing in line with the volume sold. Manufacturers 

outside the EU must provide chemical registration and toxicity data on their chemicals 

prior to receiving approval to import into the EU. In this way, the toxicity data collected 

via REACH could be shared with the US EPA to improve the enforcement of TSCA. 31 

The International Network for Environmental Compliance and Enforcement published 

the second edition of the Principles of Environmental Compliance and Enforcement 

Handbook in April 2009. 57, 58 This handbook is available in portable document format 

(PDF) and can also be accessed online at the International Police Expertise Platform 

Web site. 58, 59 This handbook gives the reader an overview of compliance and 

enforcement programs, the principles involved to make them effective, and information 

on how to build and manage compliance and enforcement programs. 

The International Lawyer, a journal published quarterly by the American Bar Association 

Section of International Law, has an annual summary of selected developments, 

conferences, reports, and publications in international environmental law from the 

previous year. Review topics generally include atmosphere and climate, marine 

conservation, international hazard management, natural resources, and international 

economy and the environment. The 2009 review also included a section on 

35, 60, 61 

international environmental litigation. 

The Environmental Investigation Agency (EIA) published “Environmental Crime–A 

Threat to Our Future” in October of 2008 and it is available online in PDF. 62 This report 

stated the definition of international environmental crime in five broad areas of offenses: 

illegal trade in wildlife, illegal trade in ozone-depleting substances, illegal transport and 

dumping of hazardous waste, illegal fishing, and illegal logging and trade in timber. 

Four case studies of crimes and five examples of successful enforcement models were 

included in the report. 

With a strong emphasis on environmental harm and an international perspective on 

environmental issues, White wrote Crimes Against Nature: Environmental Criminology 

and Ecological Justice. Published in 2008, this book gave many examples of 

environmental harm from toxic waste dumping, airborne pollutants, and contamination 

of soil and water. The author included sections on environmental regulations and law 

enforcement. 63 

Tal et al. presented the interesting results of a year-long study of environmental 

enforcement actions in Israel. In the study, the authors compared the outcomes of 

criminal enforcement for environmental violations to the outcomes of administrative 

enforcement for environmental violations. Administrative enforcement actions were 

faster, required less detailed evidence, and were perceived as being less costly than 

criminal enforcement actions. The authors did warn that the use of administrative 

actions might lead a company to determine that the cost of the fines for violations could 

be less than the cost to improve environmental controls. “Rather than creating a 

564

‘polluter pays’ dynamic, the message becomes: ‘it pays to pollute.’” 64 In contrast, the 

stigma of criminal conviction or incarceration might outweigh any financial calculation or 

monetary savings in avoiding environmental improvements. The authors came to three 

conclusions from their study of 100 criminal cases and 100 administrative actions. The 

first conclusion was that the criminal enforcement process was more effective than the 

administrative process in achieving present and future compliance. The administrative 

path did not have clear consequences and a clear timetable for delaying or ignoring 

compliance requests. The administrative system contained additional weaknesses, as 

pointed out in the authors’ second conclusion; it lacked a timely schedule and a formal 

process for followup site visits to determine compliance. This time delay could allow an 

environmental violation to continue too long before a decision is made to pursue a 

criminal enforcement action. With the third conclusion, the authors stated the 

advantage of a combined benefit and penalty approach to environmental enforcement. 

The authors discovered that when enforcement personnel could offer something to the 

regulated community, such as funds to assist in making environmental improvements, 

compliance and cooperation was more likely to occur. 64 

Stott gave a brief summary of how environmental enforcement is handled in the United 

Kingdom and provided a table of the acts of Parliament and the statutes used by the 

Environment Agency to protect the environment. Stott mentioned that, in general, the 

fines for water quality violations were higher than those for waste violations and 

speculated that the difference might be due to the inability to demonstrate the real cost 

of illegal waste dumping to the courts. One example he presented was a case in which 

a man abandoned nearly 200 drums of toxic waste. The fine for this violation was about 

half of what the man had been paid when he was hired to do the “disposal.” The 

authorities ended up spending more than five times the amount of the fine in cleanup 

costs. Stott pointed out that, with new regulations, the Environmental Damage 

(Prevention and Remediation) Regulations 2008, the courts could ensure that pollution 

will be cleaned up and the costs or damages would be recovered. 65 

Bostan et al. outlined how environmental crimes are categorized in Romania. The 

authors presented the punishment categories (range of time for imprisonment or range 

of criminal fines) and listed below each of the penalties descriptions of qualifying 

violations (for example, releasing pollutants into the water, atmosphere, or soil). 

Personal responsibility for environmental crimes and the classifications of crimes 

stipulated in environmental law in Romania also were presented in the paper. 34 

Two papers, both filled with references, discussed the problems and some possible 

solutions to environmental pollution issues faced by China. Sitaraman provided the 

history behind and development of environmental laws in China. Included in the paper 

was a table listing the major environmental laws of China from 1979 to 2004 and a table 

listing the major environmental treaties signed or ratified by China. Sitaraman observed 

that the environmental laws are not always specific enough to avoid variations in 

interpretation between different regions in China. The author suggested cleanup efforts, 

advanced technology, and citizen involvement as tools that could be used in improving 

environmental protection efforts. 66 

565

Goelz suggested that a special environmental court, modeled after the New Zealand 

Environmental Court (NZEC), could provide a good start to dealing with China’s 

environmental problems. Goelz compared this proposition to the current existence of 

China’s other specialized courts, including military courts, railway transport courts, 

maritime transport courts, and forestry courts. The author made some comparisons 

between the structure and operation of the NZEC and that of the Chinese maritime 

courts and outlined how an environmental court could be designed to align with the 

Chinese legal system and best serve the goal of environmental protection. 67 

3. Environmental forensics 

3.1 10 years of environmental forensics 

March 2000 hailed the first print issue of the journal Environmental Forensics. The first 

editorial traced the emergence of “environmental forensics” back nearly 20 years to the 

need to distinguish between different petroleum hydrocarbons in the environment. 68 

Since that time, the editors noted, the field of environmental forensics had expanded to 

include a wide range of scientific techniques. An investigator was no longer just 

involved with identification of hydrocarbon products (often termed “chemical 

fingerprinting”) or the fate and transport modeling of hydrocarbons in the environment. 

Now, the editors pointed out, an investigator could use a toolbox of techniques from 

historical sources and scientific disciplines: historical aerial photography, industrial 

archeology, regulatory history, analytical chemistry, atmospheric chemistry, 

geochemistry, toxicology, hydrogeology, environmental fate and transport, computer 

modeling, and health risk assessment. 68 

That editorial also included the prediction that environmental forensics would take a 

larger role in regulatory and legal actions in the future. Environmental attorneys, 

regulatory agencies, and insurance companies seeking to establish or allocate liability 

for environmental contamination would need environmental forensics. Also, scientists 

and the courtroom would need the knowledge to distinguish between the evidence and 

expert opinions based on valid scientific methods and the unwanted intrusion of junk 

science. 68 

In the 10 years since that first editorial, Environmental Forensics has become a primary 

journal for developments in the field of environmental forensics. It also has been one of 

the best sources for reviews on the disciplines and techniques used in environmental 

forensics. A newcomer to this type of investigative work would find many excellent 

fundamental reviews throughout this journal. The legal and investigative emphasis of 

Environmental Forensics complements two other journals in the environmental arena, 

the Royal Society of Chemistry’s Journal of Environmental Monitoring and the 

International Journal of Environmental Analytical Chemistry, the official journal of the 

69, 70 

International Association of Environmental Analytical Chemistry. 

566

Morrison published a two-part critical review of environmental forensics techniques in 

the journal during its first year. 71, 72 Morrison stated that when the results of 

environmental forensics techniques are introduced in the courtroom as evidence, the 

scientific validity of the data can be examined and challenged, often with success. His 

purpose for writing the review papers was to give an overview of the different 

techniques available and the benefits and purpose of each. The reader could then 

choose the single technique or combination of techniques needed to successfully 

investigate and prove the case. 71, 72 Table 1 lists the topics from these two Morrison 

review papers. It also lists other important review papers published in Environmental 

Forensics and the key topics from those papers. 

A wide variety of topics have been published in Environmental Forensics since 2007. 

Table 2 lists a survey of the topics in these categories. These topics have been 

organized in the general categories of contaminants, techniques, location or media, 

legal, or other. 

Table 1. Topics in Selected Environmental Forensics Review Papers 

Title, Author(s) and Year Topics 

Critical Review of Environmental 

Forensic Techniques, Part I 71 

• Aerial Photography 

• Underground Storage Tank Corrosion Models 

RD Morrison 

• Commercial Availability of a Chemical 

2000 

• Chemical Formulations and Applications Unique to a 

Manufacturing Activity 

• Polychlorinated Biphenyls (PCBs) 

• Age Dating Chlorinated Solvents 

• Presence of Non-Aqueous Phase Liquids (NAPLs) 

Critical Review of Environmental 

Forensic Techniques, Part II 72 

• Petroleum Hydrocarbons 

• Fingerprinting 

RD Morrison 

• Proprietary Additives 

2000 

• Alkyl-leads 

• Oxygenates 

• Dyes 

• Stable Isotope Analysis 

• Weathering and Biomarkers 

• Degradation Models 

• Pristane/Phytane Ratios 

• Benzene/Toluene/Ethylbenzene/Xylene (BTEX) 

Ratios 

• Contaminant Transport Models 

567



Application of Forensic 

Techniques for Age Dating and 

Source Identification in 

Environmental Litigation 73 

• Underground Storage Tank Corrosion Models 

• Commercial Availability of a Chemical 

• Chemical Applications Unique to a Manufacturing 

Activity 

RD Morrison 

• Age Dating and Source Identification of Chlorinated 

2000 

Solvents 

• Age Dating and Source Identification of Petroleum 

Hydrocarbons 

• Proprietary Additives 

• Alkyl-Leads 

• Lead Scavengers 

• Oxygenates 

• Stable Isotope Analysis 

• Biomarkers 

• Degradation Models 

• Contaminant Transport Models 

• Contaminant Transport through Pavement and Soil 

• Ground Water Models 

Application of Aerial 

Photography and 

Photogrammetry in 

Environmental Forensic 

Investigations 74 

• Photograph Acquisition 

• Interpretation of Aerial Photography 

WM Grip, RW Grip, and RD 

Morrison 

2000 

• Trial Presentation of Aerial Photography 

Age Dating of Environmental 

Organic Residues 75 

• Long-Term Methods: 

o Radiometric Carbon-14 ( 

IR Kaplan 

2003 

14 C) 

o Amino Acid Racemization 

• Short-Term Method: 

o Release Times of Crude Oil, Refined 

Petroleum Fuels, and Chlorinated Solvents 

Fingerprinting of Hydrocarbon 

Fuel Contaminants: Literature 

Review 76 

• Fingerprinting Techniques 

• Characterization of Hydrocarbon Fuel Contaminants: 

o Fingerprinting of Fuel Contaminants Using 

H Alimi, T Ertel, and B Schug 

Polycyclic Aromatic Hydrocarbon (PAH) 

2003 

Compounds 

o Fingerprinting of Fuel Contaminants Using 

Biomarkers 

• Fate of Hydrocarbon Fuels in the Environment: 

o Weathering of Volatile Aromatic Hydrocarbons 

o Weathering of n-Alkanes (or n-Paraffins) 

o Weathering of PAHs 

o Weathering of Biomarkers 

568



Combustion-Derived Polycyclic 

Aromatic Hydrocarbons in the 

Environment—A Review 77 

• Formation of PAHs 

• Environmental Fate: 

o Physicochemical Properties 

ALC Lima, JW Farrington, and CM o Biodegradation 

Reddy 

o Photodegradation and Chemical Oxidation 

2005 

• Source Apportionment: 

o Source Diagnostic Ratios 

o Historical Records 

o Stable Carbon Isotopic Composition 

o Radiocarbon Measurements 

Forensic Fingerprinting of 

Biomarkers for Oil Spill 

Characterization and Source 

Identification 78 

• Biomarker Chemistry 

• Biomarker Analysis Methodologies 

• Characteristic Fragment Ions of Biomarkers 

• Biomarker Distributions 

Z Wang, SA Stout, and M Fingas • Diagnostic Ratios (Indices) and Cross-Plots of 

2006 

Biomarkers 

• Unique Biomarker Compounds 

• Effects of Weathering on Chemical Composition of Oil 

and Biomarkers 

• Application of Multivariate Statistical Methods for 

Biomarker Fingerprinting 

Applied Dendroecology and 

Environmental Forensics. 

Characterizing and Age Dating 

Environmental Releases: 

Fundamentals and Case 

Studies 79 

• Fundamentals: 

o Scientific Background 

o Dendroecological Principles 

o Field and Laboratory Procedures 

o Ring-Width Data and Principles 

o Characterizing the Environmental Release 

J-C Balouet, G Oudijk, KT Smith, I • Case Studies: 

Petrisor, H Grudd, and B 

o Contamination by Heating Oil 

Stocklassa 

o Chlorinated Solvent Plume 


o Unknown Plumes 

o Leaking Underground Storage Tanks 

o Former Gasoline Service Station 

Age Dating Heating Oil Releases, 

Part 1. Heating-Oil Composition 

and Subsurface Weathering 80 

• Heating-Oil Composition 

• Heating Oil in the Subsurface 

• Assessing Heating-Oil Release 

G Oudijk 

• Factors Influencing Petroleum Weathering 

2009 

o Hydrocarbon Physical State, Petroleum 

Chemistry 

o Temperature, Light 

o Contact with Water, Hydrologic Conditions 

o Soil Lithology, Texture, and Moisture 

o Oxygen and Nutrients, Redox Conditions 

o Vegetation, Bacteriocides 

• Sequence of Biodegradation 

• Christensen and Larsen Method 

569



Age Dating Heating Oil Releases, 

Part 2. Assessing Weathering 

and Release Time Frames 

Through Chemistry, Geology and 

Site History 81 

• Age-Dating Methodology 

• Assessing Petroleum Weathering with 

Chromatograms 

• Sampling and Laboratory Analyses 

• Evaluating the Age Range 

G Oudijk 

• Critiquing the Matrix Age Range 

2009 

• Range of Error 

• Case Study 

The Rise and Fall of 

Organometallic Additives in 

Automotive Gasoline 82 

• Tetraethyl Lead-Based Additives 

• Lead Scavengers 

• Tetraethyl Lead Extenders 

G Oudijk 

• Mixed Lead Additives 


• Lead Phase-Out 

• Organometallic Anti-Knock Agents 

Reconstructed Plume Method for 

Identifying Sources of 

Chlorinated Solvents 83 

• Degradation Pathways 

• Reconstructing Chlorinated Solvent Plumes 

• Factoring in Groundwater Flow Direction: A Case 

BL Murphy and F Mohsen 

Study 


• Trichloroethane (TCA) Plume Age-Dating 

Table 2. Survey of Recent Topics Published in Environmental Forensics (2007–May 

2010) 

Category Topics Include 

Contaminant Polychlorinated biphenyls, lead, benzene, chlorinated hydrocarbons, 

gasolines, fuels, pesticides, dioxins, methyl tert-butyl ether (MTBE), 

polycyclic aromatic hydrocarbons, air particulates, mercury 

Technique Fingerprinting (hydrocarbons, lead), ground-penetrating radar (GPR), 

environmental modeling, wipe samples and analysis, carbon aerosol 

measurements, microscopy, statistics, dendrochronology, isotopic 

Location or 

Media 

characterization 

Surface water, ground water, river water, public drinking water, coastal 

erosion, soils, air, nutrient concentrations, sewage overflow, mining waste, 

buried waste 

Legal Bias and testimony, Daubert, cost allocations, crime investigation 

Other Biological infections, climate change, greenhouse gases, chemical warfare 

agent surrogates, standard reference materials (SRM), radiological hazards 

570

3.2 recent books and papers 

Hester and Harrison edited the 2008 book Environmental Forensics. This was volume 

26 in the Royal Society of Chemistry Publishing series, “Issues in Environmental 

Science and Technology.” Contributors to this book tackled topics of source 

identification, microbial techniques, spatial considerations of stable isotope analysis, 

chemical fingerprinting of petroleum, chlorinated solvents, and ground water pollution. 84 

Wang and Stout compiled the expertise of more than 30 scientists into the 2007 book 

Oil Spill Environmental Forensics: Fingerprinting and Source Identification. Topics from 

the 17 chapters included methods and factors affecting petroleum fingerprinting, spill 

site investigation, petroleum biomarker fingerprinting, identification by comprehensive 

two-dimensional gas chromatography, quantitative chemical fingerprinting, chemical 

heterogeneity of modern marine fuel oils, biodegradation of oil hydrocarbons, oil spill 

remote sensing, and case studies. 85 

Murphy and Morrison released a second edition of their book, Introduction to 

Environmental Forensics, in 2007. This book was designed to give the reader an 

organized presentation of the forensic tools available to use in environmental forensics. 

Updates from the first edition included chapters on laser ablation inductively coupled 

mass spectrometry (LA-ICPMS), scanning electron microscope (SEM) techniques, Xray 

diffraction (XRD), pattern recognition methods, and emerging forensic techniques. 

The chapters on sampling techniques and statistical methods were also expanded for 

the second edition. 86 

Mudge was the editor of Methods in Environmental Forensics, published in 2009. 

Mudge also authored or co-authored four of the chapters. Topics in this book included 

radionuclides, chemical fingerprinting of petroleum hydrocarbons, biomarkers and 

stable isotopes, volatile organic compound (VOC) analysis, molecular microbiology, 

statistical methods, air pollution monitoring, and evidence and expert witnesses. 87 

Mudge also contributed a chapter in Criminal and Environmental Soil Forensics. 88 Even 

though this book had a heavy emphasis on soil forensics specific to the effects from 

cadavers and footwear, it contained some chapters that could be useful for the analysis 

of pollutants in soils. These chapters included discussions on tracing soil and ground 

water pollution, current and future spectroscopic methods of analysis for soils, and a 

novel handheld sensor using anodic stripping voltammetry for real-time detection of 

heavy metals. 89 

Selected papers published on environmental forensics topics are listed and briefly 

described in Table 3. Most of these papers are from the past three years, but a few 

older review papers are included as an aid to the interested reader. 

571

Table 3. Recent Papers in the Field of Environmental Forensics 

Category Description 

Contaminant • Use of three methods (ground water modeling, subsurface environmental conditions, and isotopic fingerprinting) to 

evaluate the extent of dissolved perchlorate from multiple sources 90 

• Discussion and case study on analysis and attribution of polycyclic aromatic hydrocarbons in sediments; emphasized 

need for analysis that includes both parent (non-alkylated) and alkylated compounds 91 

• Investigation of polychlorinated biphenyl concentrations in gas and particles from ambient air samples taken in a suburb 

in Bursa, Turkey; temperature, evaporation, and atmospheric transport were factored into the study 92 

• Study of the differences in chemical composition of used and unused motor oils; used motor oils showed the presence of 

gasoline combustion residues and PAHs 93 

• Testing at an oil and gas processing facility for oil characteristics that would distinguish between naturally-occurring crude 

oil seepage and oils present on the site due to anthropogenic causes; use of gas chromatography/mass spectrometry 

(GC/MS) and isotopic techniques to determine chemical signatures 94 

• Characterization of ambient air particulate mass and ionic species near industrial zones and the Taiwan Strait in central 

Taiwan; major components of fine (PM2.5) and coarse (PM2.5-10) particulate matter were identified along with possible 

sources 95 

• Characterization and identification problems of two oil spills (off-road diesel and crude oil) undergoing methanogenic 

biodegradation; concerns expressed about fingerprinting during this degradation process 96 

• Physical and chemical characterization of samples of “slag” and “tar-like” materials; physical analysis included density, 

microscopic character, magnetic properties, and float-sink behavior; chemical analysis included the chemical 

fingerprinting of the total extractable hydrocarbons, polycyclic aromatic hydrocarbons, and petroleum biomarkers 97 

• Study on methods to identify sources of PAHs in soils; analyzed for 45 PAHs, measured PAH ratios, and measured nalkanes; 

recommended the extended 45 PAH analysis before pursuing additional methods for source identification 98 

• Total alkylated PAH characterization using Ion Signature software 99 

Technique • Use of satellite images to detect sources of pollution on the coast of Lebanon; more than 80 major sources of pollution 

were detected using the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) and Landsat 7 

ETM+ satellite images 100 

• Use of moderate resolution imaging spectroradiometer (MODIS) remote sensing technique to take spectral 

measurements of five oil types (kerosene, lubricating oil, light diesel oil, heavy diesel oil, and crude oil); MODIS also used 

to monitor Jeyeh, Lebanon, storage tank leaking event 40 

• Distinguishing oil spill patterns on water and determining the size of a major oil spill on the coast of Lebanon using 

satellite imagery 101 

• Use of ground-penetrating radar and electrical resistivity imaging (ERI) to assess the geometry and extent of illegally 

buried toxic waste 102 

572



• Two case studies in using geological materials to exclude or link suspects to illegal waste disposal in Northern Ireland 103 

• Study on soil-sampling techniques using soil treated with fipronil insecticide and placed in either opaque bags that block 

ultraviolet (UV) light or clear glass jars; factors of time, temperature, humidity, and UV light exposure were considered in 

the study 104 

• Use of marker compounds and analysis by gas chromatography-mass spectrometry (GC-MS) and gas chromatography 

with flame ionization detection (GC-FID) to identify the oil contamination in the Bohai Sea of China 105 

• Use of hollow fiber based liquid-phase microextraction (HF-LPME) coupled with GC, GC-MS, and gas chromatographyisotope 

ratio mass spectrometry (GC-IRMS) to identify hydrocarbons in oil spills 44 

• Geochemical study of the tailings after processing and disposal of the Agios Filippos high sulfidation epithermal deposit 

(located in Kirki in northeast Greece); assessed the toxicity and potential for acid generation from the tailings 106 

• Use of gas chromatography-combustion-isotope ratio mass spectrometry to determine stable carbon isotopes for ethenes 

in ground water samples to understand history of a trichloroethene (TCE) ground water plume; some results pointed to a 

tetrachloroethene (PCE) contamination 107 

• Use of gas chromatography/high-resolution mass spectrometry (GC/HRMS) to identify compounds from water samples 

taken above a pollutant plume 108 

• Fingerprinting five types of commercial oils (heavy diesel, crude oil, diesel, gasoline, and lubricant) using three 

dimensional fluorescence spectroscopy and GC-MS 39 

• Use of compound-specific stable carbon isotope analysis and consideration of physical site characteristics responsible for 

petroleum weathering to understand the age of methyl tert-butyl ether releases 109 

• Characterization of PAH sources in rivers and coastal environments of Louisiana using principal components analysis 110 

• Study of arsenic contamination in Bangladesh using geographic information systems (GIS) and univariate and bivariate 

statistical analysis 111 

• Use of carbon and hydrogen isotopes to create a two-dimensional isotope fingerprint to differentiate gasoline sources in 

environmental assessments of gasoline-contaminated sites 112 

• Study on the potential for rapid semi-quantitative surface mapping and analysis of “contaminants” (tested on powdered 

aspirin and caffeine) using a direct analysis in real time (DART)/time-of-flight mass spectrometer 113 

• New GC-MS method to distinguish between anthropogenic petroleum hydrocarbons and biogenic organic compounds in 

order to properly assess contamination sites and toxicity risks 43 

• Study to correlate odor index to pollutants from a pharmaceutical plant using open path Fourier transform infrared (OP- 

FTIR) and dispersion modeling; two volatile organic compounds emitted by the plant were ethyl acetate and acetone 114 

• Comparison between two different sampling media, quartz filters and overhead projection film, for ambient air particulates 

and particulate mercury at a site in central Taiwan 115 

573



• Development of new method for compound-specific chlorine stable isotope analysis for vinyl chloride using a continuous 

flow-isotope ratio mass spectrometer (CF-IRMS); samples were taken from a confined sandy aquifer contaminated with 

vinyl chloride and located near Ferrara in northern Italy 116 

• Investigation of sources of benzene and chlorobenzene in a shallow and deep aquifer using the combination of 

compound-specific isotope analyses, hydrogeologic data, contaminant concentrations, and site history 117 

• Analysis of soil samples to determine concentrations of residual organochlorine pesticides (OCP): used gas 

chromatograph-electron capture detection (GC-ECD) and gas chromatograph-negative chemical ionization mass 

spectrometry (GC-NCI-MS) after Soxhlet extraction and Florisil solid phase extraction (SPE) column cleanup 118 

• Critical review of compound-specific stable isotope analysis of organic contaminants using gas chromatography-isotope 

ratio mass spectrometry; 2004 paper with biodegradation table and over 100 references 119 

• General review of forensic applications of isotope ratio mass spectrometry, including references to the determination of 

the origin of contaminants in the environment, analysis of environmental contaminants, and carbon and chlorine isotopes 

in chlorinated organic compounds 120 

• Review of the applications of stable isotopes to environmental studies including discussions on BTEX, PAHs, PCBs, and 

MTBE 121 

• Critical review on the current and future uses of enriched stable isotope analysis in biological systems; includes tables 

Location or 

Media 

listing tracers for aquatic ecosystems, terrestrial ecosystems, animals, and humans 122 

• Nutrient analysis of the Ria Formosa Lagoon in Portugal; nitrogen sources traced to agricultural areas, and phosphates 

traced to a golf complex and sewage discharges 123 

• Analysis of sediment samples from eight locations in the Ria Formosa Lagoon for PAH concentrations; the most 

significant source of PAHs in the summer was determined to be boat traffic 124 

• Determining water quality and sources of pollution in the Ljubljanica River in Slovenia; input of pollution was categorized 

as coming from point sources (tannery and municipal effluents) or diffuse sources 125 

• Assessment of the metal contamination in the Riou Mort River watershed, an area affected by mining and ore-treatment 

activities, using hydrological and geochemical monitoring 126 

• Sampling and analysis for organic pollutants along a portion of the Iloilo River in the Philippines; analysis focused on 

three major groups: PAHs, branched and cyclic saturated hydrocarbons, and sterols 41 

• Site assessment of a section of floodplain used for the disposal of tailings and waste from multiple milling and smelting 

operations; historical documents, aerial photographs, geochemical and stratigraphic data were used to develop mixing 

equations to quantify contributions from the different mining sources 127 

• Use of a methodology to predict the nitrogen concentration in the ground water beneath unsewered areas; field 

measurements matched predictions of nitrogen concentration; methodology was applied to the Liman Region in Antalya, 

Turkey 128 

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• Study of PAHs and hopanes in atmospheric aerosols from samples taken in nine locations in peninsular Malaysia; 

biomass burning and vehicular emissions were determined to be the main sources of PAHs, and the significant source of 

hopanes was determined to be crankcase oil in street dust 129 

• Case investigation of a subsurface plume of non-aqueous phase liquid (NAPL) at an automotive service station; four 

different gasoline formulations were identified from analysis with the conclusion that four separate gasoline releases 

occurred 130 

• Investigation of a subsurface plume of NAPL at a truck fueling facility; two separate sources of diesel contamination were 

determined from the distribution patterns of diesel-range alkane and aromatic hydrocarbons and from an evaluation of 

source and weathering diagnostic ratios 131 

• Study of a large coastal area to assign sources to PAH contamination; sources were classified as petrogenic, pyrogenic, 

diagenetic, or biogenic 42 

• Compilation of data from ambient air studies in Asia from 1997 to 2006, including sampling techniques, sampling 

instruments, sources, and average concentrations for dry deposition and related metallic elements 132 

• Study of total gaseous mercury concentrations in indoor and outdoor air in residential locations in downtown Chongqing, 

China; results indicated higher concentrations at night and during the summer, but gaseous mercury concentrations were 

highly variable overall due to anthropogenic emissions, atmospheric changes, and unique events 133 

• Review of methyl tert-butyl ether water monitoring data from drinking water supplies across the United States 134 

• Characterization of street sediments in Kolkata, India; samples were taken from roadsides in residential, industrial, and 

inner city areas and from five sites along a highway; results of energy-dispersive X-ray fluorescence (EDXRF) analysis 

showed higher concentrations of copper, zinc, tin, lead, antimony, chromium, and nickel than from ambient or control 

soils; Kolkata metals concentrations were compared to reported analyses of street sediments from other major cities 135 

Legal • Advice for expert witnesses on presenting scientific findings in a Daubert hearing 136 

• Calculating cost allocations to account for plume scenarios and petroleum contamination in soil and ground water 137 

Other • Discussion on background values in environmental forensics, including references for general forensic techniques 

(historical document review, fingerprinting, statistical analysis) and specific forensic techniques (isotopic fingerprinting, 

geophysical method for evaluating metal backgrounds, dendroecology, and deoxyribonucleic acid [DNA] fingerprinting) 138 

• Discussion of an error in the 1,1,1-trichloroethane (1,1,1-TCA) degradation pathway; reports the literature history with 

both correct and incorrect degradation pathways 139 

• Study of natural spring water from the Eastern Black Sea region of Turkey of the activity concentrations of four 

radionuclides ( 226 Ra, 232 Th, 40 K, and 137 Cs); concentrations of the elements in water were measured using inductively 

coupled plasma-optical emission spectroscopy 140 

• Review of developments in standard reference materials (SRMs) for environmental forensics, including details on 

recertified and reissued standards and new standards for organic contaminants and biological and environmental 

matrices 141 

575



• Discussion of the defective Chinese drywall used in thousands of homes in the United States that off-gas sulfur 

compounds; overview of the scientific and legal aspects of the problem 45 

• Editorial on age-dating gasoline spills when insufficient or inadequate sampling has already occurred; includes table on 

various plume observations that can give an indication of the age of the plume 142 

• Crime investigation of a landfill site containing hazardous waste from tannery and shoe factories located in Arroio do 

Meio, Rio Grande do Sul, Brazil; traditional crime scene examination conducted, along with a detailed description of the 

area; examination of possible pollutant sources; historical data gathering and sampling; and analysis of water, sediment, 

and animal tissues 143 

576

4. Electronic waste 

Electronic waste or “e-waste” has been the fastest-growing part of the solid waste 

stream since the mid-1990s. 144 In 2004, an estimated 100 million personal computers 

worldwide entered the waste stream for either recycling or disposal. 145 In 2005, more 

than 40 million computers were discarded in the US alone. 146 Governmental agencies, 

multinational committees, and environmental conservation groups internationally are on 

4, 5, 11, 147-153 

alert to find solutions to this immense waste problem. 

Personal computers (including central processing units, monitors, keyboards and 

peripherals) are not the only type of waste electrical and electronic equipment (WEEE) 

of concern. Electrical and electronic equipment (EEE) that might be discarded into the 

waste stream includes large and small household appliances, cellular and regular 

telephones, portable digital assistants (PDAs), video and audio equipment (televisions, 

stereos, cameras, video recorders), electrical tools, and other computer equipment 

9, 154, 155 

(scanners, printers, computer game units). 

Some discarded EEE is classified as used or UEEE for the purpose of reuse and/or 

recycling. WEEE or e-waste may be banned from importation into a country, but the 

same country may allow the importation of UEEE as part of a reuse or recycling 

operation. Li et al. listed the criteria to distinguish between WEEE and UEEE for 11 

Asian countries, but the criteria are not uniform. The authors stated that the 

development of consistent standards could help restrict illegal transboundary movement 

and processing of e-waste. 156 

Trade shipments of UEEE frequently include some WEEE. The quantity of WEEE in 

the shipment of UEEE can be an amount negotiated between the exporter and the 

recycler/importer. As part of a deal for desirable UEEE, the recycler/importer agrees to 

deal with disposing of the useless WEEE. Or, the addition and amount of WEEE 

included in a shipment of UEEE can be an unfortunate surprise. Either way, the WEEE 

must be disposed of somehow in the importing country. Schmidt presented the findings 

of a Basel Action Network (BAN) investigation in Africa, with examples of the 

unregulated EEE market at work. 157 BAN representatives observed thousands of 

vendors in the Ikeja Computer Village, near Lagos, Nigeria, selling repaired and 

refurbished electronics such as computers, fax machines, and cell phones; these were 

products from the UEEE imports. They also observed piles of e-wastes filling swamps; 

these were the EEE junk items, the WEEE. When the piles of e-waste grew too high, 

the piles were burned, producing fumes that affected the local residents. BAN 

estimated that, of the purported UEEE shipped to Nigeria, between 25 to 75 percent 

was just waste or WEEE. 157 

Williams et al. presented the multiple facets to the reuse and recycling of computers in 

the global market and described some contrasting points of view about the e-waste 

issue. The authors used published research to make the argument that the hazards of 

leaching toxic materials out of landfills are minimal and that disposing of old computers 

in landfills could be environmentally preferable to recycling. In contrast, though, a lack 

577

of old, reusable computers would decrease the ability for poorer, developing markets to 

access information technology through low-priced, refurbished computers. 

Economically, the authors stated, the reuse and recycling market provides employment, 

but unregulated recycling efforts have caused great harm to the environment and 

human health. 158 

Ogunseitan et al. described a major difficulty in dealing with the international e-waste 

problem in their 2009 policy brief. The difficulty is the worldwide patchwork of 

regulations and standards dealing with three parts of the e-waste problem: the 

movement of hazardous wastes, the use of hazardous substances, and the recycling 

responsibilities of the EEE producers. The authors explained that uneven 

implementation of standards had created “risk holes” in poor and developing 

communities. These are areas where markets trading in second-hand electronics and 

the recycling of illegally imported or domestic e-waste thrive. 144 Mckenna expressed 

concerns about illegal e-waste in developing areas and discussed the application of the 

Basel Convention and WEEE Directive to the e-waste problem. 159 The Basel 

Convention set into action a multi-national agreement on the control of transboundary 

movements and disposal of hazardous wastes. 6 The WEEE Directive of the European 

Parliament (Directive 2002/96/EC) set an objective to reduce the quantity of waste 

electrical and electronic equipment and the harm to the environment. 9 Together, these 

European initiatives have the potential to decrease illegal e-waste. 

5. Regulatory and legislative considerations 

Ladou and Lovegrove briefly outlined the e-waste problem and presented the 

regulations and initiatives that might assist countries in dealing with e-wastes. The 

Basel Convention was described first, followed by the European Restriction on 

Hazardous Substances (RoHS) and the WEEE legislation. The authors mentioned the 

US EPA RCRA regulations but indicated that, even with the openness of the US EPA to 

consider improvements to these regulations, many impediments exist to actually 

revising the regulations. It was noted that several US states have passed or developed 

regulations similar to the RoHS legislation of Europe. Regulations developed in Asia 

such as the Home Appliance Recycling Law (HARL, Japan) and the Law for Promotion 

of Effective Utilization of Resources (LPEUR, Japan) have promoted the recycling of 

millions of appliances per year. The Environmental Protection Agency of Taiwan 

assigned the responsibility of recycling waste personal computers to producers in 1997. 

The authors added that several Asian countries are actively working on their own ewaste 

recycling or RoHS-type regulations. 160 

The US Government Accountability Office produced a report in 2008 on the weakness 

of the US regulations to address e-waste exports. The report explained that cathode 

ray tubes (CRT) are the only electronic device regulated by the US EPA. In order for 

CRTs to be exported for reuse, the US EPA must receive a notification of the intent to 

export CRTs and approval from the importing country. The report gave examples of the 

578

need for increased enforcement of the RCRA regulations limiting CRT exports, and it 

recommended expanding the scope of the CRT rule to include other e-waste types. 13-15 

Kutz provided a quick review of the e-waste problem, including a section on examples 

of what initiatives are being taken to correct the problem. The author described the 

efforts of businesses, the restrictions on hazardous chemicals as directed by the EU, 

and the possibilities of biodegradable and eco-friendly (non-plastic) case designs. 

Programs in Japan, Canada, and Taiwan were listed as examples that the US could 

observe in order to design a program that could best address e-waste issues. Two 

programs were emphasized as potential e-waste solutions: advance recovery fees 

(ARFs) and extended producer responsibility (EPR). With ARFs, consumers pay a 

recycling or recovery fee for the electronic item at the time the item is purchased with 

the purpose of collecting enough funds to support e-recycling programs. EPR programs 

can place responsibilities on manufacturers (or producers) with the purpose of having 

the manufacturer cover the costs of managing the waste at the end of the product life. 

Kutz recommended an EPR program for the US and provided guidance for structuring 

that system. 161 

EPR and WEEE management initiatives were discussed by Nnorom and Osibanjo. The 

authors listed examples of the administrative, economic, and informative policy 

instruments that could be used to implement EPR. Administrative instruments included 

implementing product take-back programs, setting emission limits, and setting reuse 

and recycling targets. Economic approaches included advance disposal fees and 

deposit-refund systems. Environmental reports and labeling were two of the informative 

instruments listed. WEEE management initiatives in the EU, Finland, Japan, 

Switzerland, and Taiwan were also described. 162 

Krishna and Kulshrestha discussed the problem of e-waste in developing nations and 

focused their attention on Indian and Chinese directives. The authors mentioned the 

Basel Convention and the EU WEEE and RoHS Directives and also described the 

Bamako Convention. The Bamako Convention was adopted by 51 African countries 

and banned the import and trafficking of hazardous waste into Africa. 163 

E-waste initiatives for China, Nigeria, and India were the topics of several other papers. 

Ye et al. discussed the elements of the current legislative framework for e-waste 

management in China and analyzed the challenges that still exist. 12 Ni and Zeng 

presented a set of recommendations to address the e-waste crisis that included 

enhancing law enforcement, establishing national or local e-waste disposal centers, and 

coordinating with other countries on e-waste. 164 Umesi and Onyia suggested solutions 

for e-waste problems in Nigeria and lamented the fact that the Basel Convention 

provisions are not being enforced in originating countries. The authors recommended 

establishing adequate material collection systems and additional systems for the proper 

reuse, refurbishment, recovery, and recycling of materials; this plan would create 

employment and provide environmentally sound management. 155 Nnorom and 

Osibanjo reviewed the flow of secondhand and scrap electronic devices into Nigeria and 

noted that much of the growth in the information and communications technology in 

579

Nigeria had come from secondhand electronics. Management practices for e-waste 

were discussed, and the recommendation was made for formal recycling facilities and 

the implementation of a plan to check secondhand EEE for functionality before 

importing the devices. 165 Bandyopadhyay reviewed the e-waste management initiatives 

in India and included tables on the economics of the e-waste trade, estimates of ewaste 

in Mumbai, and the Indian WEEE policy framework. 166 Dwivedy and Mittal 

estimated current and future quantities of e-waste generation in India. Estimates of ewaste 

quantities could help recyclers make decisions in planning recycling infrastructure 

and capacity building. Total WEEE estimates for 2007-2011 were given at 

approximately 2.5 million metric tons, about 30 percent of which would be from personal 

computers. 167 

6. Contamination and health effects 

Materials used to manufacture electrical and electronic equipment include both valuable 

and hazardous components. Unregulated recovery of the valuable materials, such as 

copper wire or gold from UEEE or WEEE, can lead to situations in which very 

hazardous components are released, which may endanger human health and the 

environment. In his recent review on e-waste, Robinson made the point that the 

chemical composition of e-waste varied by the age and the type of the discarded item. 

The author stated that the composition continually changes to meet the demands of 

technological developments, the regulatory requirements of environmental agencies, 

and the requests of environmental organizations. 168 A table of environmental 

contaminants found in e-waste was provided in Robinson’s review, and the health 

effects of many of these contaminants were listed by Schmidt. 169 

Basel Action Network representatives visited a city in China that was, and still is, a 

center for e-waste metals recovery. 169 Residents of Guiyu, according to BAN, had been 

receiving e-waste to process since 1995. The processes used for recovering metals 

included no protection for human health or the environment. Residents heated circuit 

boards in woks to melt lead solder to remove computer chips; the molten lead and 

burning plastic created toxic fumes. The molten lead was then poured on the ground, 

contaminating the soil and nearby river area. Pits filled with “aqua regia,” a strong 

combination of hydrochloric and nitric acid that can produce smoke and fumes, were 

used to dissolve the computer chips. Used acid and sludge were dumped into the local 

river. 169 

Guiyu was visited in 2008 by BAN and a crew from the CBS network’s primetime 

investigative show, 60 Minutes. The show obtained footage of smoldering piles of ewaste; 

residents heating circuit boards in the open air and pooling molten lead; 

residents using a large acid bath for gold recovery; and views of the smoky air and the 

dark, polluted river water. 170-172 

Fresh water samples from inside and outside of Guiyu were analyzed for metals by 

Wong et al. High metal concentrations attributable to the acid leaching of e-wastes 

(silver, cadmium, copper, and nickel) were found. Lead isotope composition from river 

580

samples was studied and indicated that lead from multiple sources was present in the 

two local rivers. 19 Air, soil, and sediment samples from locations in Guiyu were 

analyzed for persistent organic pollutants (flame retardants, dioxins and furans, 

polycyclic aromatic hydrocarbons, and polychlorinated biphenyls) and heavy metals in 

another study. Open combustion of e-waste and the dumping of materials at the end of 

the recycling process were suspected as major sources of toxic chemicals in the local 

environment. As expected, results from air analyses indicated high levels of the 

persistent organic pollutants and heavy metals when compared to samples from other 

cities. Soil and sediment analysis results also indicated extremely high levels of 

pollution. 173 

Lead and cadmium levels in the blood of the children living in Guiyu were the focus of 

work by Zheng et al. The authors sampled blood from children in Guiyu and from 

children in nearby Chendian, a town with no e-waste processing but with clothing 

manufacturing as the primary industry. Analyses found that children in Guiyu had 

higher levels of lead and cadmium in the blood when compared to the control children of 

Chendian. 174 Xing et al. tested for PCB levels in local river fish, air, and human milk 

samples in Guiyu. Results indicated that inhalation exposure was a more significant 

contributor to PCB levels in humans than dietary exposure (fish). 20 

Analyses were performed on soil samples taken in e-waste recycling areas in the 

province of Taizhou, China. Jun-hui and Hang sampled the rice paddy soils from seven 

different sites, all with past or present e-waste recycling operations. The authors found 

soil samples from one of the active e-waste recycling sites to be high in cadmium. 16 

Tang et al. analyzed soil samples taken from Wenling, an emerging e-waste recycling 

city in Taizhou. The samples were analyzed for heavy metals (copper, chromium, 

cadmium, lead, zinc, mercury, and arsenic) and persistent organic compounds, 

including polycyclic aromatic hydrocarbons and polychlorinated biphenyls. Results 

indicated that most heavy metals exceeded the Grade II value of soil quality standards 

set by the State Environmental Protection Administration of China. PCB levels ranged 

from 2 to more than 200 times higher in the e-waste area soil samples than in samples 

taken from reference sites. PAH concentrations were higher in the samples taken near 

small, household workshops. Overall, the authors found the small, household e-waste 

recycling workshops to be greater contributors to local soil contamination than largescale 

plants. 18 

Samples from an e-waste recycling facility, agricultural land, and a chemical-industrial 

complex in Taizhou were analyzed for polybrominated dibenzo-p-dioxins and 

dibenzofurans (PBDD/Fs) by Ma et al. The matrices included workshop-floor dust, 

electronic shredder residues, leaves from trees and shrubs, and surface soils. 

Estimated daily human intakes of PBDD/Fs were calculated. 175 Chen et al. sampled 

surface sediment of the Nanguan River running through Taizhou. The authors 

determined the concentrations of PCBs, PAHs, and heavy metals in the samples. 

Toxicity studies were also performed. The authors noted that pollutant and toxicity 

levels near industrial recycling parks were much lower than areas near household 

581

workshops. This was attributed to the management and recycling technologies at use 

in the industrial parks. 176 

Wang analyzed scalp hair samples to determine heavy metal exposure to residents in 

Taizhou and used samples from two control sites for comparison. Lead, copper, 

manganese, and cadmium were significantly higher in concentration in the samples 

from Taizhou than in the control samples. 177 

Contamination by e-waste recycling in Bangalore, India, was studied by Ha et al. The 

authors sampled soil, airborne particulates, and human hair from e-waste recycling sites 

and reference sites in Bangalore and Chennai and analyzed the samples for trace 

elements. Concentrations were reported for 16 elements (vanadium, chromium, 

manganese, cobalt, copper, zinc, molybdenum, silver, cadmium, indium, tin, antimony, 

mercury, thallium, lead, and bismuth). In each results table, the analytical data from this 

study were compared to metals analysis data from other published studies. The 

authors concluded that a link could be made to metal contamination and exposure from 

e-waste recycling. But, the authors conceded, since not all sample types were taken 

from every site and no lifestyle considerations were factored into the hair analysis, this 

study was only a preliminary start for further investigations. 17 

7. Analytical methods 

Testing for the hazards posed by e-waste in the US has centered on the Toxicity 

Characteristic Leaching Procedure in the US EPA Test Methods for Evaluating Solid 

Waste, Physical/Chemical Methods (SW-846). 178 This test attempts to mimic conditions 

for waste items in a landfill. Liquid in a landfill may contact waste and may cause the 

leaching of metals or organic chemicals from that waste into the liquid. A breech in the 

landfill structure could allow this liquid to exit and contact soils, surface waters, or 

ground water. If one or more of 40 contaminants exceed toxicity limits, then the waste 

may be considered to be hazardous and issues of waste treatment and land disposal 

restrictions may need to be addressed. 179 Questions about the metals that might leach 

from e-waste disposed in landfills have led to several studies using computer parts and 

the TCLP method. 

The primary researcher and research center for e-waste and TCLP during the past 10 

years has been Townsend at the University of Florida in Gainesville. 169 The work 

published by his research group has shown that lead is the main concern for toxic, 

leachable metals in e-waste. Publications by the group are listed and described in 

Table 4. Townsend and his group also produced two reports, one for the Florida Center 

for Solid and Hazardous Waste Management on lead leachability from CRTs using the 

TCLP method, and another for the US EPA in 2004 on RCRA toxicity characterization of 

180, 181 

computer central processing units (CPU) and additional electronic devices. 

Lincoln et al. tested cellular telephones for metals and organic compounds using three 

methods: the US EPA TCLP, the California Department of Toxic Substance Control 

(DTSC) Waste Extraction Test (WET) and California DTSC’s Total Threshold Limit 

582

Concentrations (TTLC). Phones were grouped by physical dimensions; phones outside 

a specific size range were eliminated. Phones were shredded, homogenized, then 

divided equally by mass. The authors determined that the phones exceeded TCLP 

limits for lead and failed the standards of the TTLC for five metals (copper, nickel, lead, 

antimny, and zinc). No limits were exceeded in the WET test. 22 

Keith et al. used four different leachability tests on a selection of computer CRTs, 

printed circuit boards, computer mice, television (TV) remote controls, and mobile 

phones. Each of the six devices were crushed and then tested using the US EPA 

TCLP, the US EPA Synthetic Precipitation Leaching Procedure (SPLP, Method 1312), 

the Dutch Environmental Agency Availability Test (EA NEN 7371), and method DIN S4 

of the Institut fur Normung, Germany. The extracts were analyzed for lead, cadmium, 

chromium, and silver. TCLP extractions were repeated on composite CRT glass (one 

additional extraction) and filter papers with residual solids (two additional extractions), 

and then those extracts were filtered and analyzed. Results indicated increased lead 

leachability with smaller CRT and circuit board particle sizes and extracted lead 

increased with the repeated TCLP extractions. The SPLP, NEN 7371, and DIN S4 tests 

yielded very low results for the leachable metals, and the authors did not recommend 

those tests for assessing potential metal leachability from e-waste. 21 

Researchers are actively seeking ways to detect unwanted chemicals in EEE 

fabrication, ways to better deconstruct EEE for recovery or reuse of materials, and ways 

to destroy or isolate the usable and disposable WEEE. This requires a thorough 

understanding of the chemical composition of the devices, and most researchers have 

provided the chemical analysis process details and data in their published work. Lee 

and Wright developed a standard operating procedure for assessing and confirming 

compliance with regulations on the use of ozone-depleting chemicals. 23 Cui and 

Forssberg used a variety of methods to characterize television scrap and determine 

cost-effective ways to separate the components of a shredded TV mix for potential 

material recovery. 182 Lee et al. presented a good overview of the recycling process for 

scrap computers, including tables on compositions of components, separation methods, 

and CRT coating-removal methods. 183 Méar et al. characterized the waste funnel and 

panel glass from dismantled CRTs in order to develop reuse applications. 184 Chen et al. 

tested a pyrovacuum process that removes and recovers lead from the funnel glass of 

CRTs and makes a porous glass residue. 185 Guo et al. reviewed the recycling methods 

for the non-metallic fractions of printed circuit boards. 186 Hino et al. developed a 

method for pulverizing waste printed circuit boards with integrated circuits. 187 Niu and Li 

studied two methods for making printed wire boards non-hazardous: high-pressure 

compaction and cement solidification. The low-impact resistance of the compacted 

samples made them too unstable to be a long-term disposal solution. The results from 

the TCLP tests on the highly impact-resistant, solidified samples showed lead at 

concentrations well below the regulatory limits; a good indication that this method could 

render the boards non-hazardous. 188 

583

Table 4. Publications from the Townsend Research Group, University of Florida, 

Gainesville 

Title, Author (s), Year Description of Research and Results 

Characterization of Lead 

Leachability from Cathode 

Ray Tubes Using the Toxicity 

Characteristic Leaching 

Procedure 189 

SE Musson, Y-C Jang, TG 

Townsend, and I-H Chung 

2000 

Leaching of Lead from 

Computer Printed Wire 

Boards and Cathode Ray 

Tubes by Municipal Solid 

Waste Landfill Leachates 190 

Y-C Jang and TG Townsend 

2003 

RCRA Toxicity 

Characterization of Discarded 

Electronic Devices 191 

SE Musson, KN Vann, Y-C 

Jang, S Mutha, A Jordan, B 

Pearson, and TG Townsend 

2006 

Factors Affecting TCLP Lead 

Leachability from Computer 

CPUs 192 

KN Vann, SE Musson, and TG 

Townsend 

2006 

Evaluation of a Modified 

TCLP Methodology for RCRA 

Toxicity Characterization of 

Computer CPUs 194 

K Vann, S Musson, and T 

Townsend 

2006 

Used TCLP procedure on 36 CRTs from television and 

computer monitors; found that most color monitors (21 of 

30) exceeded regulatory limits for lead, but no 

monochrome monitors exceeded lead limits; the frit seal 

in color CRTs was a major source of lead 

Varied the leachates used to test printed wire boards and 

CRTs from television and computer monitors; test 

methods and leachates used were the standard TCLP 

method; leachates that were sampled from 11 Florida 

landfills, California’s Waste Extraction Test, and the US 

EPA Synthetic Precipitation Leaching Procedure; 

leaching test results indicated that lead concentrations 

were highest with the TCLP test, followed by WET, then 

leaching with the landfill leachates, and, lastly, the SPLP 

test 

Thirteen electronic devices (printers, cell phones, 

laptops, flat-panel displays, computers peripherals, etc.) 

were tested using a standard TCLP test and/or a 

modified TCLP test; tests were modified to be either 

large-scale or small-scale; all toxicity characteristic 

metals were below regulatory limits except lead; a 

correlation was observed between the amounts of 

ferrous metal in the device and lead concentrations in the 

leachate 

Performed TCLP particle reduction on computer central 

processing units, then leached different combinations of 

the CPU components; high lead concentrations were 

observed when leaching printed wire boards; again 

observed the correlation between decreased lead 

concentrations when ferrous metal was included and 

discussed the connection of iron and zinc concentrations 

and the hydrous ferric oxide interaction as previously 

reported by Kendall; 193 also noted increased lead 

concentration with increased head space and that 

additional particle size reduction below the size required 

for TCLP did not increase the lead concentration 

Large-size leaching modification to TCLP test to allow 

disassembled CPUs to be leached almost whole in a 

large vessel in order to be more representative of the 

device and decrease sample preparation time; observed 

higher lead concentrations for the large-scale leaching 

when compared to standard TCLP conditions; reported 

that the large-scale leaching did reduce sample 

processing time (sample particle reduction), but 

584

Table 4. Publications from the Townsend Research Group, University of Florida, 

Gainesville 

Title, Author (s), Year Description of Research and Results 

Impact of Electronic Waste 

Disposal on Lead 

Concentrations in Landfill 

Leachate 195 

E Spalvins, B Dubey, and T 

Townsend 

2008 

8. Biological waste 

increased the cost of analysis 

Created simulated landfills in large columns using a 

manufactured municipal solid waste (MSW) in five 

columns and excavated waste and soil in the sixth 

column; identical sets of e-waste were added to three of 

the manufactured MSW columns, and dechlorinated tap 

water was added to all columns on nearly 30 occasions; 

analysis of extracts from the columns indicated lead 

concentrations between columns with e-waste were not 

statistically significant when compared with lead 

concentrations in the control columns 

8.1 Concentrated animal feeding operations (cafos) 

Animal Feeding Operations (AFOs) are defined as agricultural operations where 

animals are kept and raised in a confined area. 196 Animals live and grow in a 

production setting that contains feed, excrement, and sometimes dead animals, all in a 

limited space. The US EPA has defined AFOs by animal confinement times and the 

lack of natural vegetation in the confined area. Concentrated Animal Feeding 

Operations (CAFOs) must meet the definition of an AFO, and, in addition, CAFOs must 

meet one of the specific regulatory definitions for small, medium, or large CAFOs. 

CAFOs may be regulated by the US EPA under the National Pollutant Discharge 

Elimination System (NPDES) which is a part of the Clean Water Act. 196-198 

8.1.1 Environmental Impacts 

CAFOs are a concern in the environment due to the large amounts of waste produced 

from the confined animal productions. Potential problems include leaking, leaching, or 

runoff from manure stockpiles or storage pits, which can contaminate ground water and 

fresh water sources. 199, 200 Water contamination is not the sole concern; emissions of 

gases and particulate matter from the production facilities may cause harm to human 

health. 

Water Environment Research publishes several annual reviews on the types of 

pollutants that can contaminate surface or ground water. Several review papers 

address the contaminants that can originate from agricultural waste and the detection of 

chemicals or pathogens in the environment. 201-209 

Lee et al. prepared an in-depth review of the effect of agricultural antimicrobials and 

hormones on soil and surface and ground waters. The authors covered the use and 

occurrence, sorption, degradation, transport processes, and ecological and health 

effects of the chemicals. Analytical methods and suggestions for future research were 

585

also included. The authors stated that more studies were needed to make conclusive 

connections between animal-derived antibiotic-resistant pathogens and human health 

impacts. 210 

Dolliver and Gupta addressed the potential for manure runoff to be a source of 

emerging contaminants to the environment. The authors quantified three commonly 

used beef antibiotics, chlortetracycline, monensin, and tylosin, in runoff from beef 

manure stockpiles. Due to concerns from the spread of antibiotic resistance in the 

environment, they suggested covered facilities to decrease runoff. The authors also 

compared the antibiotic runoff concentrations to the antibiotic concentrations reported in 

studies about composting and manure-applied fields; the latter concentrations were 

generally found to be much less than those found in runoff losses. 199 

Sapkota et al. sampled surface and ground water up-gradient and down-gradient of a 

swine facility to determine if antibiotic-resistant enterococci and other fecal indicators 

were present. The results indicated higher concentrations of fecal bacteria in the down 

gradient surface and ground water samples. The down-gradient surface water 

concentrations were consistently in excess of the US EPA bacterial water quality for 

recreational standards. 200 

Seepage of animal wastes stored and treated in anaerobic lagoons can lead to an 

increase of nitrate concentrations in the ground water. Excess nitrates in rivers can 

lead to areas of hypoxia, or oxygen deficiency, which can dramatically affect the plant 

and fish life. Mariappan et al. used nitrogen isotope ratios in their study to identify 

sources of lagoon seepage. 15 N isotope enrichment was dependent on the ambient 

temperatures and the rates of mixing or addition of fresh wastes. 211 

Burkholder et al. reviewed the possible contaminants in CAFO waste and the potential 

impacts to water and wildlife. 212 The authors asserted that current livestock waste 

management practices are inadequate to protect water resources. This review was 

prepared as part of the mini-monograph, “Environmental Health Impacts of 

Concentrated Animal Feeding Operations: Anticipating Hazards—Searching for 

Solutions,” which resulted from a 2004 scientific conference and workshop addressing 

CAFO environmental health issues. 213 

Emissions from CAFOs can be more than just malodorous and can include ammonia, 

hydrogen sulfide, and particulate matter contaminated with endotoxins. Endotoxins are 

inflammatory agents produced by bacteria that can either aid in the development of 

immunity or cause harm by inducing asthmatic reactions. Heederik et al. reviewed the 

human health effects from CAFO emissions as part of the CAFO mini-monograph 

mentioned above. 214 

Wash and runoff water from CAFOs can be used beneficially as a fertilizer and soil 

215, 216 

amendment when an acceptable Nutrient Management Plan (NMP) is followed. 

An additional advantage is the reduction in the need for fresh water irrigation in fields 

when the CAFO water is applied. 

586

Bradford et al. provided a thorough review on the use of CAFO wastewater on 

agricultural lands. The review was divided into three sections: environmental 

contaminants, land application, and treatments. Six classes of potential waste lagoon 

contaminants were addressed in the environmental contaminant section: nutrients and 

organics, heavy metals, salts, pathogens, antibiotics, and hormones. For each of these 

classes, the authors described the potential environmental problems. For example, the 

authors stated that some waste lagoon water exceeded the recommended copper and 

zinc concentrations for irrigation water. The potential negative effects from an excess of 

these metals could be phytotoxicity and surface and ground water contamination. The 

land application section of the review discussed the US EPA requirement for NMPs that 

are designed to consider all nutrient input sources, nutrient losses, and nutrient uptake 

rates for the plants. 215 The authors explained the basic assumption of the NMP, that all 

CAFO contaminants would be absorbed or degraded in the root zone and no 

contaminants would enter the surface or ground water. Possible weaknesses to NMP 

designs and implementation also were discussed in the paper. 216 

8.1.2 Analytical Methods for Emissions 

Bunton et al. reviewed methods for ammonia and hydrogen sulfide monitoring. The 

review contained information on the type of equipment and analyzers that have been 

used in various studies. The authors provided cautionary summaries regarding odor 

measurements and the monitoring of particulate matter and described potential 

problems with each. For odor measurements, a chemical characterization can be 

performed and constituents quantified, but the result may be a complex mixture of 

compounds and the correlation of the compounds to human scent responses can be 

poor. Also, taking only air samples in the field may leave a study incomplete because 

some odorous material may attach to particulate matter that could be filtered out, 

depending on the sampling method. For particulate matter emitted from CAFOs, 

instrumentation does exist for sampling and measurement, but the authors pointed out 

that dust in the rural areas around CAFOs has the potential to interfere with those 

results. The authors also included sections on plume dispersion models and 

recommendations for future studies. 217 

Ni and Heber provided an extensive review of ammonia sampling and measurement 

techniques for animal facilities in volume 98 of the Advances in Agronomy series. The 

review included sampling methods categorized into closed, point, and open-path 

methods and advice for selecting a method. Measurement techniques and devices 

were described in brief and were well-referenced. A comparison of ammonia 

measurement devices and summaries from several comparative studies were provided 

at the end of the measurement section. An important addition to the review was the 

detailed table of potential sources of error in the sampling or measurement of ammonia 

concentrations. The authors categorized the errors (calibration, sampling, 

measurement, and data processing) and provided suggestions to reduce or eliminate 

each source of error. 218 

587

Todd et al. used acid gas washing samplers and a calibrated flow injection analyzer to 

obtain ammonia concentrations from sampling sites at a beef cattle feedyard. 

Continuous ammonia measurements were taken during a summer period using a 

chemiluminescence analyzer. Measurements for wind velocity and direction, air 

temperature and humidity, and precipitation were also collected to provide further data 

for modeling ammonia emission rates in spring, summer, and winter months. 219 

Trabue et al. measured volatile sulfur compounds at swine and poultry feeding 

operations. They employed a canister-based method and used fused silica-lined minicanisters 

to collect the air samples at lagoon and building locations. The sampled air 

was focused into a gas chromatograph system with both mass spectrometer and pulsed 

flame photometric detectors (PFPD). Target compounds for the analysis were 

hydrogen sulfide, carbonyl sulfide, methanethiol, dimethyl sulfide, carbon disulfide, 

dimethyl disulfide, and dimethyl trisulfide. Storage stability for samples was a problem 

especially when sampled from warm, humid environments. 220 

8.1.3 Methods for Tracking Waste Releases 

Several research groups have published their efforts to find one or more optimal 

methods to trace the source of an overflow or leak from a CAFO. 28, 199, 200, 211 Causes of 

leaks or overflows can be obvious, for example, from a weather related event (heavy 

rains filling an uncovered lagoon) or a structural or engineering fault (a lagoon wall 

failure). Observed lagoon problems like these likely would not require tracking methods 

to find the source of waste contamination and encourage a CAFO operator to improve 

the facilities to avoid future contamination events. But, when fecal environmental 

contamination occurs and several potential sources of fecal waste exist, using source 

identification tools may be the solution to track down the cause of contamination. 

Environmental contamination from fecal waste is not limited to AFOs or CAFOs. When 

urban areas are involved, fecal material entering the environment may come from 

humans (leaks in sanitary sewers or sewer connections to storm drains), domestic 

animals (including pets), and urban wildlife (birds, raccoons, rats, etc.). Mallin et al. 

listed these contributors to waste from urban areas and studied the impacts from 

stormwater runoff in urban, suburban, and rural areas. The authors noted that, in some 

rural areas, unsuitable septic systems, pastureland, and farmland receiving applications 

of manure as fertilizer can contribute to fecal material found in waterways. 26 

Chemicals such as artificial sweeteners, pharmaceuticals, detergents, fragrances, and 

caffeine can be used to trace discharges into a waste stream. 30, 221 Buerge et al. in 

Switzerland sampled untreated and treated wastewater, ground water, and tap water 

and tested for the presence of four sweeteners, acesulfame, cyclamate, saccharin, and 

sucralose. The authors observed that acesulfame was prevalent in all waters and was 

not removed through wastewater treatment, but cyclamate and saccharin were 

removed. Buerge’s team concluded that assumptions could be made about the source 

of a leak from those sweeteners. Assuming that some contamination has occurred, 

ground water samples without saccharin and cyclamate could point to a treated 

588

wastewater leak and ground water samples with those two sweeteners could indicate a 

leak from untreated wastewater. 221 

Using specific chemicals to locate the source of a release of fecal waste into water 

resources is one of the tracking methods discussed in the literature and, in the paper by 

Cimenti et al., it is listed as “chemical microbial source tracking” or chemical MST. 

Microbial source tracking encompasses many different methods that can be used alone 

for a single-source investigation or in combination for scenarios in which multiple 

sources of contamination exist. The large volume of research and developments in 

MST is complex, but some excellent papers exist to help the investigator decide the 

most efficient and informative tracking approach. 30 

8.2 Microbial source tracking 

The goal of microbial source tracking is to determine the source of a fecal contamination 

event. To trace back to the origin, it is necessary to determine what species produced 

the fecal material. Is the fecal material of human or animal origin? If animal, can 

indicators point to the type of animal (agricultural animals, domestic animals, urban 

wildlife)? 

Contamination can also be from multiple sources. More than one method of tracking 

may be needed to locate those sources and the pathways used to spread the fecal 

waste in the environment. Considering the ecology of an area can be an essential tool 

in understanding the spread of contamination. 25 

Cimenti et al. provided an excellent overview of microbial source tracking in their 

concise, informative review paper. The authors acknowledged several fundamental 

papers in microbial source tracking in their text. 222-225 In the paper, MST methods were 

sorted into five classifications: direct monitoring of pathogens, culturing methods, 

phenotypic methods, genetic methods, and chemical methods. The classifications and 

several examples of each were listed in a summary table and later described in detail 

with methods and references to published work. The authors suggested that direct 

monitoring of pathogens, while effective for determining health risks, may not be the 

most efficient or affordable. More than one pathogen may exist in water affected by 

fecal contamination, and detection of pathogens can be complex and costly. 30 

The other four method classes (culturing methods, phenotypic methods, genetic 

methods, and chemical methods) track fecal indicators, which, if chosen well, should 

only be present whenever fecal contamination is present; the indicators should not be a 

normal part of the environment. An ideal indicator should also be effective for tracking 

in all types of waters (surface, ground, and marine waters). These indicator 

characteristics and a few others (fast analysis, cost-effective, etc.) were listed in the 

Cimenti paper and credited to a book by Maier et al. 226 Gerba contributed the chapter 

on indicator microorganisms, and in it he listed the criteria for an ideal indicator 

organism. 227 

589

Detection of enteroviruses falls into the category of direct monitoring of pathogens. 

Enteroviruses are very host-specific, and detection of these confirms fecal 

contamination. But, the authors pointed out, the lack of enteroviruses in a sample does 

not exclude fecal contamination because water polluted by feces does not always have 

enteroviruses present. 30 Rajtar et al. detailed the danger of enteroviruses in the aquatic 

environment and listed some methods of detection. 228 

Culturing methods involve isolating a microorganism that is specific to a potential source 

of the fecal pollution (humans, agricultural animals, domestic animals, urban wildlife). 

These microorganisms can include fecal coliforms, fecal streptococci, protozoa, or 

viruses. The authors cautioned that culturing media is rarely selective and that few 

organisms are host-specific and have “ideal indicator” criteria as presented in Maier et 

30, 226, 227 

al. 

Phenotypic methods look at bacterial species that are present in both humans and 

animals, but differentiate the bacterium based on how it changed while existing within a 

specific host. Different phenotypic characteristics develop under different intestinal 

conditions. The authors stated that biochemical responses to these methods could be 

too similar to distinguish a difference, but, if multiple phenotypic characteristics are 

detected, the accuracy of the method could be improved. Antibiotic resistance analysis 

is one phenotypic method. Bacteria are isolated, cultured, and measured for their 

responses to several antibiotics. 30 

The preferred MST methods are the genetic methods. These use intestinal bacteria to 

discern sources of fecal pollution by either making a host-specific match for DNA or 

identifying a genetic characteristic that indicates an adaptation to a specific host. The 

drawback to the genetic method is that it requires databases or libraries to match the 

genetic profiles, and these profiles can vary by location and time. The authors provided 

brief descriptions of a few genetic methods, but point to other papers for more 

information. 30 

Cimenti et al. also listed several possible chemical tracers that can be either directly 

associated with fecal material or can be discharged in the same wastewaters as fecal 

material. These chemical methods, the authors pointed out, do not easily correlate to 

pathogens, but can still help in locating a source of pollution. Using information about 

emerging contaminants in water, the authors provided lists of veterinary and human 

antibiotics, veterinary medicines, prescription and non-prescription drugs, and 

hormones as possible chemical tracers. 30 

Field and Samadpour published a review of source tracking methods and included 

details on the applicability of certain methods, the usefulness of the methods, and the 

drawbacks of the methods. They expressed concerns about public health and the 

occurrence of pathogens in water and suggested a combination of targeted pathogen 

monitoring with targeted fecal source tracking to better regulate water quality. 229 Zhang 

et al. reviewed antibiotic resistance genes and provided detailed tables, categorized by 

590

antibiotic, that list the genes, the biological and environmental sources, and references 

of papers with studies on each specific gene. 230 

Stoeckel and Hardwood prepared a review of MST that addressed the strategies for 

developing, validating, and evaluating a microbial source tracking study. The authors 

compared method performance data, including statistics, and outlined considerations for 

the field study design. 231 Santo Domingo et al. reviewed MST methods and listed the 

research gaps and suggested some goals for future studies. 27 

Witty et al. addressed the complexity of determining a single source of pollution from 

multiple potential and contributing sources. The authors used multiple methods but 

concluded that DNA sequence analysis for MST in combination with a study of the 

ecosystem is a better approach than microbiological applications alone. 25 Reischer et 

al. emphasized the need to consider hydrological catchment dynamics in source 

tracking study design and gathered data for their study over a 31-month period of 

time. 232 Stapleton et al. performed a catchment scale study and found that MST 

techniques needed further development in order to become a useful tool in catchment 

systems. 233 Vogel et al. studied water and sediment samples in a catchment reach for 

fecal contamination. The authors found Escherichia coli in both water and sediment, 

while the Bacteroidales markers were detected more frequently in the sediment 

samples. 234 

Wolf et al. created a library-independent “viral toolbox” (VTB) to distinguish between 

human and animal fecal pollution. Their VTB used real-time reverse transcription (RT)polymerase 

chain reaction (PCR) assays for norovirus genogroups, adenovirus species, 

and F+ RNA bacteriophage genogroups. The authors found their assays to be highly 

sensitive and specific to the targets. 29 

Microbial source tracking papers are filled with PCR assays. Agudelo et al. used a 

multiplex real-time PCR assay for fast quantification and obtained higher detection 

levels than with conventional microbiological techniques. 235 Kortbaoui et al. used dotblotting 

membranes containing specific oligonucleotides for human and for four animals 

(bovine, porcine, ovine, and chicken) to detect mitochondrial DNA in water samples. 

The dot-blot assays were found to be as specific and sensitive as a nested-PCR 

approach. 28 Lamendella et al. evaluated two swine-specific PCR assays. The 

Prevotella strain-based PCR assay provided positive results more often than the 

methanogen-based PCR assay, and it also had a high number of cross-reactions with 

non-target fecal DNA samples. 236 

591

9. Acknowledgments 

This paper would not have been possible without the assistance of several people, and I 

offer my thanks to them. Thank you to Nancy Greer for her constant help with acquiring 

the books and publications needed for this review paper and for responding quickly and 

knowledgably to all the miscellaneous queries I sent her way. 

Thank you to Diane Sipe and Beth Mishalanie for their critical reviews of the entire 

paper; their questions and corrections provided clarity and readability in many sections. 

Thank you to Mark Goodwin and Jennifer Lee for their comments to improve my 

accuracy and presentation of the information in the biological section. 

Thank you to Lindsay Powis for entering more than 200 references into the EndNote 

bibliography software. And, thank you to Dianne Clark for providing her editorial check 

of the review paper and preparing it for outside distribution. 


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607

DRUGS AND TOXICOLOGY  

  608 

Toxicology 

Review : 2007 - 2010 

Compiled by 

Wing-chi Cheng Ph.D.; Wing-sum Chan Ph.D.; Nai-leung Poon Ph.D. 

Government Laboratory, Hong Kong 

CHINA 

Presented by 

Fu-chiu Kwok Ph.D. 

All correspondence should be addressed to Dr. Fu-chiu Kwok, Acting Assistant 

Government Chemist, Government Laboratory, 7/F, Homantin Government 

Offices, 88 Chung Hau Street, Ho Man Tin, Kowloon, Hong Kong, China. 

609


  

1. INTRODUCTION  613  

2. CURRENT TOXICOLOGICAL ISSUES  613  

2.1 Driving Under the Influence  613  

2.1.1 Alcohol  613  

2.1.2 Drugs  616  

2.1.2.1 Detection of DUID  616  

(i) Impairment Approach  616  

(ii) Drug Presence Approach  617  

2.1.2.2 Epidemiological studies  621  

2.2 Drug Facilitated Sexual Assaults (DFSA)  625  

2.3 Workplace Drug Testing  627  

2.3.1 Urine  628  

2.3.2 Alternative specimens  631  

2.3.2.1 Oral fluid  631  

2.3.2.2 Hair  632  

2.3.2.3 Sweat  634  

2.4 Emergence of New Drugs of Abuse  635  

2.4.1 Phenethylamines  635  

2.4.2 Piperazines  636  

2.4.3 Synthetic cannabinoids  637  

2.4.4 Pyrrolidinophenones  638  

2.5 Survey on Trends of Drug Use  638  

2.5.1 Opiates  638  

2.5.2 Amphetamines  639  

2.5.3 GHB  640  

2.5.4 Ketamine  640  

2.5.5 Other Drugs  641  

2.6 Quality Assurance and Accreditation  643  

610

3. ADVANCES IN TOXICOLOGICAL ANALYSIS 645  

3.1 Instrumental techniques  645  

3.1.1 GC‐MS  645  

3.1.2 LC‐MS  646  

3.1.3 Capillary electrophoresis  648  

3.2 Extraction Techniques  649  

3.2.1 Liquid‐phase microextraction (LPME)  649  

3.2.2 Electrokinetic membrane extraction (EME)  650  

3.2.3 Solid‐phase extractions (SPE)  650  

3.2.4 Microextraction in packed sorbent (MEPS)  651  

3.2.5 Solid‐phase microextraction (SPME)  651  

3.3 Analysis of Specific Drugs  652  

3.3.1 Markers for ethanol consumption  652  

3.3.2 Small molecules  653  

3.3.3 Drugs of abuse  653  

3.3.4 Chemical warfare agents  655  

3.3.5 Toxic herbs and venoms  657  

3.3.6 Drugs in sports and doping control  658  

3.3.6.1 Anabolic androgenic steroids  660  

3.3.6.2 Stimulants  662  

3.3.6.3 Synthetic protein hormone, blood doping and gene doping  662  

3.4 ALTERNATIVE SPECIMENS  665  

3.4.1 Oral fluid  666  

3.4.2 Hair  667  

3.4.3 Sweat  671  

3.4.4 Meconium  672  

3.4.5 Other specimens  673  

3.5 Interpretation of toxicological results  674  

3.5.1 Drug levels in postmortem specimens  674  

3.5.1.1 Alcohols and related biomarkers  675  

3.5.1.2 Abused drugs  677  

3.5.1.3 Other drugs  679  

3.5.2 Drug stability, artifact and post‐mortem redistribution  682  

3.5.3 Pharmacogenomics  684  

611

4. CONCLUSIONS                          685  

5. REFERENCES                          688  

612

1. INTRODUCTION    

Forensic toxicology is of paramount importance to the judicial system and for 

legal protection of individuals and society, in providing scientifically sound 

information relevant to the cause of incident. Areas of interest include, but are 

not limited to, road safety, criminal activities with suspected involvement of 

drugs/poisons, workplace drug testing, drugs of abuse monitoring in prisons 

and custodial medicine, death investigations, and doping in sports. 

Scientific advancement on one hand facilitates the development of more 

sensitive and specific toxicological examination techniques for detection of a 

wider scope of drugs/poisons at very low concentrations in biological 

specimens that would have otherwise been difficult or impossible in the past. 

On the other hand, it also helps in filling in the gaps to better understand the 

toxicological findings through research such as controlled experiments, 

epidemiological studies, and compilation of casework results of similar types. 

Scientific advancement, however, also results in the development of new 

drugs, both in the form of pharmaceutical preparations and illicit drugs, which 

pose continual challenges to toxicologists with respect to developing strategies 

for their identification, quantification, and interpretation of results. This review 

is divided into 2 parts, namely “Current Toxicological Issues” and “Advances in 

Toxicological Analysis”. It covers the most significant progress in toxicology 

published over the past 3 years. 

2. Current Toxicological Issues 

2.1 Driving Under the Influence 

Driving is a very complex task, requiring simultaneous coordination of several 

different cognitive and psychomotor functions. Although crashes can be the 

consequence of many different factors, driving under the influence of alcohol 

and drugs is considered to be one of the major problems worldwide, causing a 

million people to be killed and many millions more to be injured in traffic 

crashes. 

2.1.1 Alcohol 

Driving under the influence of alcohol (DUI) is continued to be one of the major 

road safety problems. The breath alcohol concentration (BrAC) has been used 

extensively in many jurisdictions for DUI enforcement and prosecution 

613

purposes. The pulmonary exchange processes of ethyl alcohol after drinking 

were studied [1,2]. During a single-exhalation breath test, a person with a 

smaller lung size can give a greater BrAC reading [1]. With 504 pairs of BrAC 

and BAC (Blood Alcohol Concentration) tests evaluated, the mean hourly 

BrAC elimination rate was found to be 0.082 mg/L/h (0.050–0.114, 95% range) 

[3]. Paired BAC and BrAC (BAC, in g/dL, and BrAC, in g/210 L), were 

determined for 11,837 drivers apprehended in New Zealand. With a mean time 

delay of 0.73 h between breath and blood sampling, the BrAC false-positive 

rate decreased from 31.3% to only 2.8% after time-adjustment for BAC using 

0.019 g/dL/h as the blood alcohol clearance rate [4]. In a controlled study 

performed with volunteers who drank 3 glasses of wine, 46% of females and 

no male subjects exceeded a BrAC of 0.25 mg/L. The main predictor of 

self-perception of impaired driving skills was the feeling of nausea, followed by 

a BrAC in excess of 0.25 mg/L [5]. The usefulness of portable, handheld 

breath analysers was assessed by re-calculation of the results using mean 

elimination rates of 0.156 ±0.099 g/L/h for blood and 0.091± 0.054 mg/L/h for 

breath; the readings from the portable instruments were in very good 

agreement with the results taken from stationary devices, but correlation with 

the results of blood analysis was weaker [6]. 

Some drivers may claim to have consumed alcohol after traffic accidents, 

causing difficulties in prosecution. The relationships of the change in urinary 

alcohol concentrations (UACs) between two successive voids, and that 

between the ratio of BAC and UAC in DUI cases, were studied [7]. With 

supporting evidence from after-drink cases, the UAC/BAC ratio for the first 

void was close to or less than unity, and the UAC increased by 0.21 g/L 

between the two voids (about 1 hour interval). For those cases without 

evidence of an after-drink, the ratio was 1.46 and the UAC decreased by 0.25 

g/L on average, indicating it is in the post-absorptive phase. The inter- and 

intra-subject variability in the elimination rate and absorption rate was 

estimated with BAC measured over time in 12 male volunteers on 4 occasions. 

Results showed that inter-subject variability was substantially smaller than 

intra-subject variability when the dose varied [ 8 ]. The gender specific 

elimination rates were studied and determined that the elimination rates for 

women are generally higher than those for men [9,10]; this can partly, but not 

completely, be explained by progesterone levels [9]. A BAC analysis was 

carried out for two consecutive blood specimens to correlate the elimination 

rate of alcohol and BAC. Results showed a slight change in alcohol elimination 

614

ate with increase of BAC, with the mean value of 0.221 ± 0.075 g/kg [11]. 

The results of a pilot study have shown that the use of alcohol biomarkers 

[carbohydrate-deficient transferrin (CDT), and the Early Detection of Alcohol 

Consumption (EDAC) test] in the U.S. could provide an objective tool to 

identify high-risk drivers [12]. While a study in Canada showed that all 

biomarkers except fatty acid ethyl esters (FAEE) [phosphatidyl ethanol (PETH), 

CDT and gamma glutamyltransferase (GGT) in blood samples; ethyl 

glucuronide (EtG) and ethyl sulphate (EtS) in urine samples; and FAEE and 

EtG in hair samples] were related significantly to the interlock BAC test profiles 

[13]. Hence, it was suggested that the alcohol biomarkers should serve as a 

good means for determining license regranting for DUI offenders. The value of 

analyzing ethanol metabolites as measures of abstinence was evaluated for a 

subject who was convicted of DUI but claimed abstinence from alcohol after 

supervised disulfiram treatment [14]. Apart from traditional biomarkers, CDT, 

EtG and FAEE in hair, phosphatidylethanol in whole blood, and EtG and ethyl 

sulphate in urine were measured. The subject was found with EtG in distal 

parts of hair only, with FAEE concentrations typical for either moderate social 

drinking or abstinence. 

The efficacy of a fructose drink that claimed to noticeably increase the alcohol 

elimination rate was investigated using 30 volunteers under a randomized 

double-blind, placebo-controlled, cross-over study. While the soft drink may 

decrease the alcohol concentration by about 10%, the claim of fructose drinks 

to work as a 'soberade' cannot be proven from a scientific point of view [15]. 

A comparison of the performance of a portable impairment test device known 

as the Roadside Impairment Testing Apparatus (RITA) with the Field 

Impairment Tests (FIT) was conducted using 122 volunteers who received a 

dose of alcohol and placebo on separate days [16]. The accuracy of RITA was 

between 66 and 70%, not significantly higher than that of FIT. Horizontal gaze 

nystagmus, not currently included in FIT, was also investigated and posted an 

accuracy of 74%. Driving is a classic example of visually guided behavior in 

which the eyes move before some other action. A study showed that the 

optimal eye-hand coordination relationship deteriorates with levels of breath 

alcohol well within the current UK legal limit for driving, and that deterioration in 

eye movement can be detected by an automated in-car system, which warns 

the driver when he is no longer fit to drive [17]. 

615

2.1.2 Drugs 

Driving under the influence of drugs (DUID) is scientifically and legally more 

complex than DUI. The complexity of drug impaired driving is compounded by 

drug-alcohol or drug-drug interactions which can be additive, synergistic or 

antagonistic in nature. A major problem in assessing the true public health 

impact of drug-use on driving and overall traffic safety is that the variables 

being measured across studies vary significantly. A project was initiated by 

several international organizations to develop guidelines for research on 

drugged driving [18]. This Guidelines Document is divided into three major 

sections, each focusing upon different aspects of drugged driving research 

within the critical issue areas of 'behavior', 'epidemiology', and 'toxicology'. 

Apart from the control over the use of illicit drugs, concerns on the impact of 

the use of centrally acting medicinal drugs on driving cannot be ignored. The 

International Council of Alcohol and Drugs on Traffic Safety (ICADTS) has 

proposed a three-tier categorization system by comparing the effects within the 

three categories with effect of different blood alcohol concentrations (BAC), 

where categories I, II, and III as respectively equivalent to BACs < 0.5 g/L 

( 0.8 g/L (>0.08%) [19]. 

2.1.2.1 Detection of DUID 

Two approaches, namely the impairment approach and the drug presence 

approach, are currently used to identify DUID drivers. 

(i) Impairment Approach 

Owing to limited scopes of drugs that can be identified by drug testing devices, 

the impairment approach for law enforcement to identify DUID drivers is 

currently used in many countries. The impairment tests are based on the 

evaluation of a variety of observable signs and symptoms, and divided 

attention tests that have been proven to be reliable indicators of impairment. 

There are no roadside testing devices commercially available for the detection 

of ketamine, which is increasingly being abused in many Asian countries. A 

study found that more than 70% of ketamine abusers failed in the FIT 

assessment. The typical observable signs and symptoms being a lack of 

convergence, horizontal gaze nystamus, elevated pulse rate, and general 

failing the divided attention tests, especially the walk-and-turn and one-leg 

616

stand [20]. In Norway, an impairment-based law is used to handle suspected 

drugged driving and it was determined in a study that benzodiazepines were 

the most frequently detected drugs in comparison with other common illegal 

drugs such as Δ 9 -tetrahydrocannabinol (THC) and amphetamine [21]. The 

relationship between the blood concentration of benzodiazepines and the 

influence on performance in field sobriety tests was studied for 171 cases. The 

observations of behavior, walking, walking after turn, and Romberg's test were 

found to be significantly related to the benzodiazepines concentration, while no 

significant relation was found with respect to pupil size, nystagmus, or 

orientation [22]. Apart from illicit drugs, consumption of prescription drugs may 

also cause driving impairment. For example, the prevalence of gabapentin in 

impaired driving cases in the U.S. found that the mean concentration of 

gabapentin in blood from impaired driving cases (n = 137) was 8.4 ± 5.4 mg/L, 

and that the subjects may exhibit psychophysical indicators of a central 

nervous system depressant (e.g., horizontal gaze nystagmus) but with clinical 

indicators that are not consistent with depressants (e.g., dilated pupils, low 

body temperature, and elevated pulse and blood pressure) [23]. 12 drivers 

arrested for driving under the influence of dextromethorphan alone or in 

combination with chlorpheniramine revealed that blood dextromethorphan 

concentrations ranged from 150 to 1220 ng/mL and that the subjects were 

evaluated to be under the influence of a drug in the CNS-depressant category 

[24]. Six blood samples obtained from arrested DUID drivers tested positive for 

toluene, with blood concentrations of 12 to 45 mg/L, and the subjects 

displayed horizontal but not vertical nystagmus, elevated pulse and blood 

pressure, and lower body temperatures [25]. 

A statistically based approach to drug classification decisions in Drug 

Evaluation and Classification (DEC) evaluations suggested that nine clinical 

indicators significantly enhanced the prediction of drug category, including 

pulse rate, condition of the eyes and eyelids, lack of convergence, hippus, 

reaction to light, rebound dilation, systolic blood pressure, and the presence of 

injection sites [26]. 

(ii) Drug Presence Approach 

Blood is the preferred specimen for investigating impaired driving, but some 

jurisdictions allow the use of other specimens such as urine and oral fluid. One 

of the key elements in the enforcement process is the availability of rapid and 

reliable drug screening devices for on-site screening of drugged drivers, 

617

equired for immediate administrative measures. With recent progress in 

performance of the point-of-collection testing device performance, controlled 

drug administration studies, immunoassay and chromatographic methods, and 

collection devices, there is no doubt that oral fluid has an increasingly 

important role in testing for suspected DUID drivers [27]. In 2009, modification 

of the Traffic Act in Belgium was made to allow for random oral fluid drug 

testing and analysis of oral fluid samples for evidence of DUID (comparable to 

the Australian situation) [28]. Analytical techniques for detection of drugs in 

oral fluid have been reviewed with emphasis on applications used in European 

Union (EU) roadside testing projects [29]. Various on-site oral testing devices 

were evaluated [30,31,32,33]. While good results were found for amphetamine 

(Drugwipe, Oralab6), cocaine (OraLab, Dräger), and opiates (OraLab, 

Oralab6), improvements are still needed regarding the detection of cannabis 

and benzodiazepines. Efforts have been made to look for reliable devices for 

the detection of THC in oral fluid. 25 subjects, 5 free and 20 addicts from a 

heroin detoxification center with time of cannabis last smoked recorded, were 

used to evaluate the Cozart DDSV to detect cannabis in oral fluid. The 5 

drug-free subjects were negative for cannabis; however, the device could only 

detect 8 positive subjects (with THC concentrations in the buffer in the range 

15-219 ng/mL) [34]. In a more recent evaluation of three devices, namely 

Drugwipe-5(+), RapidSTAT and Drug Test 5000, these devices were concluded 

to be able to detect about 70% of all cocaine and cannabis users in a roadside 

setting. Moreover, with the lowering of the cutoff for THC for the new Drug Test 

5000 (i.e., 5 ng/mL), a sensitivity of 93% was reported [35]. 

In 2006-2008, 13 oral fluid screening devices for drugs of abuse were 

evaluated in operational police practice by 11 teams from 6 EU member states, 

as part of a work package of the EU funded integrated project “Driving under 

the Influence of Drugs, Alcohol and Medicines” (DRUID) [28]. The 13 

screening devices selected for the ROSITA II project were evaluated for 

operational aspects in two phases. At the end of the evaluation of over 2950 

tests, a total of 7 devices were considered to be promising (from a practical 

police perspective) for use during daily traffic law enforcement activities, and a 

set of Police Users Requirements and Specifications has been developed. 

Similarly, 8 on-site oral fluid drug screening devices were evaluated for their 

analytical performance in another DRUID project performed in Belgium, 

Finland, and Denmark. However, this latter evaluation concluded that none of 

the tests reached the target value of 80% for sensitivity, specificity, and 

618

accuracy for all the separate tests. The sensitivities of the cannabis and 

cocaine tests were all quite low; however, the sensitivities of the devices are 

generally better for amphetamines [36]. 

In the context of toxicological analysis of samples from DUID cases, 

immunoassays tailored for a limited number of drugs (of abuse) are usually 

applied as prescreening tests at the roadside and/or in the laboratory. However, 

many other common drugs, such as anesthetics, antidepressants, 

antiepileptics, antihistamines, newer designer drugs, herbal drugs, 

neuroleptics (antipsychotics), opioids, or sedative-hypnotics, can also impair 

drivers. The pros and cons of multianalyte single-stage and tandem gas or 

liquid chromatography-mass spectrometry procedures for the screening, 

identification, and validated quantification of such drugs in blood were 

reviewed with respect to their probable usefulness in impaired driving 

toxicology [ 37 ]. A survey conducted in the U.S. identified marijuana, 

benzodiazepines, cocaine, prescription and illicit opiates, muscle relaxants, 

amphetamines, CNS depressants, and sleep aids used as hypnotics, as being 

the most frequently encountered drugs in these cases [38]. Recommendations 

as to what specific members of these drug classes should (at a minimum) be 

tested for in the investigation of suspected DUID cases, with suggested 

analytical cutoffs for screening and confirmation of drugs in blood and urine, 

were reported. 

With the limited sample volume of oral fluid available for testing in the context 

of DUID, a comprehensive methodology is needed for simultaneous 

measurement of multiple analytes in a single aliquot; liquid 

chromatography-tandem mass spectrometry (LC-MS/MS) is the common 

method of choice. An ultra performance liquid chromatography-tandem mass 

spectrometry (UPLC-MS/MS) method was developed for detection of 29 drugs 

(including THC and benzoylecgonine) in oral fluid collected randomly and 

anonymously at the roadside from drivers in Denmark throughout 2008-2009. 

Using a solid phase extraction of 0.2 mL oral fluid, the LOQs were 0.5 µg/kg for 

all analytes. A preliminary account of oral fluid samples collected at the 

roadside showed the presence of amphetamine, cocaine, codeine, THC, 

tramadol, and zopiclone [39]. A semi-automated LC-MS/MS method for the 

simultaneous extraction, identification, and quantitation of 21 analytes in oral 

fluid was developed with LOQs ranged from 0.4 to 5 ng/mL using 0.2 mL of 

diluted oral fluid [40]. Another LC-MS/MS method for the detection and 

619

quantitation of common drugs of abuse in oral fluid, collected with Quantasil 

oral fluid collection kits, was reported with LODs/LOQs being 2-10 ng/mL [41]. 

A confirmational analysis of drugs, including morphine, cocaine, 

benzoylecgonine, and amphetamines in oral fluid by LC-MS was also reported. 

300 µL of oral fluid is required for the testing, with LODs of 0.5 – 1 ng/mL [33]. 

That the window of detection of drugs in oral fluid reflects the corresponding 

window in blood suggests that oral fluid is a specimen of choice for roadside 

testing. However, oral fluid/blood ratios vary from drug to drug, from person to 

person, and even intra-individually, posing challenges in interpretation of the 

results for oral fluid. Blood (B) and oral fluid (OF) samples collected from 

drivers suspected of DUID in Belgium, Germany, Finland, and Norway for the 

ROSITA-2 project were used to study the relationship between the oral fluid 

and the blood concentrations of drugs of abuse [42]. The ratios found in this 

study were comparable with those that were published previously, but the 

range was wider. The median of the OF:B ratios of basic drugs such as 

amphetamines, cocaine, and opiates were 4-13, 22, and 2-10 respectively. 

The ratios for benzodiazepines were very low (median OF:B ratio: 0.02-0.1) 

while that for THC was 15. Several methods for estimating the prevalence of 

high blood concentrations of THC and amphetamine in a population of drug 

users by analysing their OF suggested that dividing the drug concentration in 

OF by the OF/B regression coefficient gives an acceptable estimation of high 

blood drug concentrations, which may give valuable additional information on 

possible drug impairment [43]. The presence of THC in blood is usually 

interpreted as an indicator for recent drug use. However, a study of chronic 

cannabis users over 7 days of continuous monitored abstinence showed that 

substantial whole blood THC concentrations (0.3 ± 0.7 ng/mL) persisted 

multiple days after drug discontinuation in heavy chronic cannabis users [44]. A 

comparison of the impairment of driving-relevant skills by alcohol or cannabis 

suggested that a THC concentration in serum of 7-10 ng/mL is correlated with 

an impairment comparable to that caused by BAC of 0.05%. The per se limits 

for THC in blood appear to be the most effective approach to differentiating 

drivers who are impaired by cannabis use from those who are no longer under 

the influence [45]. The relationship between blood zopiclone and zolpidem 

concentrations and driving impairment was assessed by a clinical test, and the 

impairment was compared in that with a group of drivers with suspected 

alcohol-related impairment [46]. It is concluded that the percentage of impaired 

drivers with blood zopiclone concentrations above 130 µg/L roughly 

620

corresponded to the percentage of impaired drivers with BACs higher than 

0.1%, but a positive relationship of blood concentration and impairment was 

not found for zolpidem. Some prescription medications will metabolize into 

illicit drugs, leading to complicated interpretations. For instance, in a study 

conducted by Jones et al. revealed that when 6-acetylmorphine (6-MAM) was 

present in urine (N = 324), the mean morphine/codeine ratio in blood was 7.5, 

and this important ratio was less than unity in only two cases. This study finds 

compelling evidence that approximately 90% of apprehended drivers in 

Sweden with morphine and codeine in their blood had used heroin [47]. 

Some countries such as Sweden and Italy require offenders of DUID with their 

driving license suspended to have passed a drug test before considering 

re-issuance of the driving license [48,49]. In a pilot study of hair analysis of 

drivers previously convicted of DUID, both clients and clinicians thought that 

hair sampling was easier than urine sampling. With 99 hair samples and 198 

urine samples being collected from 84 clients during the 12-month study period, 

hair analysis identified illegal drugs on seven different occasions, whereas 

urine failed to identify any illegal drugs [48]. Another similar study on hair 

analysis of cocaine and its metabolite benzoylecgonine was also conducted. 

With relatively short half-life (

2007 in Sweden revealed that 22% of them had alcohol above the legal limit (> 

0.2 g/L), and 18% with drugs other than alcohol found (where 4.8% was found 

with illicit drugs, with amphetamine being the most common one identified at 

mean concentrations of 1.5 mg/L) [53]. A study on re-arrest rate of DUI and 

DUID in Sweden during a 4-year survey showed that 44% of individuals 

(16,277 out of 36,799 cases) re-offended 3.2 times on average, and that the 

re-arrest rate of DUID is higher than that of DUI [54]. The median BAC was 1.5 

mg/g (~ 0.15 g%) compared with 1.6-1.7 mg/g in recidivists, while 

amphetamine, the most frequently detected drug found in DUID drivers, with 

median concentration of 0.5 mg/L in the first-time offenders compared with 1.0 

mg/L found in blood samples in recidivists [54]. Also, a 5-year survey of DUID 

cases showed that amphetamines were found in 60% of the cases (15,898 of 

26,556 cases), with mean blood concentration of 1.01 mg/L for amphetamine; 

0.85 mg/L and 0.34 mg/L, for amphetamine and MA, respectively; and 0.23 

mg/L for ecstasy [55]. In a similar 5-year survey by the same group for cocaine 

and benzyolecgonine, most of the cases (574 cases) had only 

benzyolecgonine found in apprehended drivers, with mean concentrations of 

0.19 mg/L. For the remaining cases (221 cases), cocaine and 

benzyolecgonine were both found in the blood samples at mean 

concentrations of 0.076 mg/L and 0.859 mg/L, respectively [ 56 ]. Over a 

10-year period (1995-2004), between 18% and 30% of all DUID suspects had 

measurable amounts of THC in their blood (> 0.3 ng/mL), either alone or 

together with other drugs, with mean THC concentration of 2.1 ng/mL [57]. 

Zero-tolerance or limits of quantitation laws are a much more pragmatic way to 

enforce DUID legislation. Occurrences of GHB in blood samples from people 

arrested in Sweden for DUID between 1998 and 2007 were also studied. With 

the use of a cut-off concentration of 8 µg/mL, 215 cases (39%) were detected, 

with GHB only at mean blood concentrations of 91 mg/L [58]. In a similar study 

by the same group in 473 cases for DUID, the mean GHB blood concentration 

was 90 mg/L; people arrested for petty drug offenses (N=1061) were 111-118 

mg/L; and GHB-related deaths (N=33) were 190 mg/L. The signs of drug 

influence noted by arresting police officers included sedation, agitation, 

unsteady gait, slurred speech, irrational behavior, jerky body movements, 

dilated pupils, and spitting [59]. 

A study of trends of DUID in Finland showed that there was an 18-fold increase 

in DUID cases during 1977-2007 [60]. The most commonly found substances 

were benzodiazepines (75.7%), amphetamines (46.0%), cannabinoids 

622

(27.7%), and opioids (13.8%). A 7-year survey in Greece for the prevalence of 

alcohol/drugs involved in road traffic accidents indicated that there was a drop 

in the detection of alcohol from 37% in 1998-2000 to 29% in 2000-2004, and 

that cannabis, benzodiazepines, opiates, and cocaine were found in 4%, 4%, 

4%, and 1% of the total number of cases, respectively [61]. The prevalence of 

drugs in the first year after the introduction of the revision of the Swiss Traffic 

Law for DUID in 2005 was studied [62]. The most frequently encountered 

drugs in whole blood were cannabinoids (48% of total number of cases), 

ethanol (35%), cocaine (25%), opiates (10%), amphetamines (7%), 

benzodiazepines (6%), and methadone (5%). In the U.K., the drugs and 

alcohol findings for victims of 1047 cases in fatal road traffic incidents indicated 

that alcohol and cannabinoids were the most frequently detected substances 

across the victim groups, while a wide range of drugs were also detected (e.g., 

drugs of abuse, anti-convulsants, anti-histamines, anti-inflammatories, 

anti-psychotics, cardiac drugs, and over-the-counter products) [ 63 ]. 

Epidemiological data in fatal road accidents in France suggested that alcohol 

levels above the legal limit (0.5 g/L of blood) were found in 21% of all drivers 

involved in accidents (killed, injured, or unharmed). Cannabis headed the list of 

illicit drugs detected, with a prevalence of 6.8% (THC > or = 1 ng/mL) [64]. The 

prevalence of the use of psychoactive substances for 1000 truck drivers in 

France was studied by analyzing urine samples. The drugs found included 

cannabinoids (85 cases), ethanol (50), opiates (41), buprenorphine (18), 

methadone (5), benzodiazepines (4), amphetamines (3), cocaine (1), and 

6-MAM (1) [65]. A study in Austria for the period of 2003-2007 showed that 

cannabis (50%), opiates (20%), amphetamines (18%), cocaine (15%), and 

benzodiazepines (20%) were those with the highest prevalence in blood 

samples taken from drivers; a newly developed urinary road-site-test system, 

Check 24 (Protzek GmbH, Germany), with two different cut-off values for 

THCCOOH-glucuronide, was used to identify drivers under the influence of 

cannabis [66]. Post-mortem cases for amphetamine-based drugs in 1999-2004 

in the Netherlands were reviewed and compared with those of non-fatal DUID 

cases. Amphetamine-based drugs were present in 70 post-mortem cases and 

467 DUI cases. The majority of MDMA- and amphetamine-caused deaths 

occurred with blood concentrations above 1.5 and 0.80 mg/L, respectively, and 

the concentrations in drug related deaths and DUI cases overlap the range of 

fatal concentrations. Thus, the MDMA and amphetamine concentrations 

should never be used alone to establish the cause of death [ 67]. 274 

amphetamines (amphetamine, MA, MDA, MDMA and/or 

623

3,4-methylenedioxyethylamphetamine (MDE)) positive serum samples 

collected by police in Germany for roadside testing were also analysed for 

GHB by LC-MS/MS after acetonitrile protein precipitation [68]. Applying a 

cut-off concentration of 10 µg/mL, 5 samples exceeded this concentration and 

were found positive for GHB. 

In a retrospective study (2001-2005) of the blood samples from DUI drivers in 

Ontario, Canada for toxicological examinations for drugs, showed that THC 

was the most frequently encountered drug (n=18), followed by 

benzoylecgonine/cocaine (n=8), morphine (n=6), lorazepam (n=5), and 

diphenhydramine (n=4) [69]. 

The state of Victoria in Australia has had a random drug-testing program for 

drivers since 2004, targeting THC and methamphetamines (MA and MDMA) in 

oral fluid [70,71]. With 13,176 roadside drug tests performed in the first year, 

the overall drug positive rate was 2.4% of the screened population. There was 

one false positive for cannabis when the results of both on-site devices were 

considered, and four for methamphetamines [70]. Oral fluid samples were 

collected from drivers to estimate the prevalence of drug driving in Queensland, 

Australia [72]. Positive rate for at least one illicit substance was 3.7% [72]. 436 

specimens taken from injured drivers involved in motor vehicle collisions and 

subsequently transported to a major adult trauma centre in Victoria, revealed 

that metabolites of cannabis were the most commonly found drug (46.7%). 

THC was found in 33 specimens, which was almost exclusively found in the 

15-44 years age group. Also, there was an increasing use of benzodiazepines 

in older drivers, particularly women >65 years old [73]. 

1635 motor vehicle drivers were randomly selected in Thailand in 2005-2006 

to determine the prevalence of psychoactive drug and alcohol use among 

general drivers [ 74 ]. 5.5% tested positive for breath alcohol, with 2% 

exceeding the legal limit (≥ 50 mg%). A psychoactive drug was found in 158 

(9.7%) urine samples. The top 3 most frequently detected licit drugs were 

antihistamines (2.0%), sedative cough suppressants and benzodiazepines, 

while the most frequently detected illicit drugs were amphetamine (1.8%), 

cannabis (1.1%), mitragynine (Kratom) (0.9%), and morphine (0.1%). 

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2.2 Drug Facilitated Sexual Assaults (DFSA) 

The true prevalence of DFSA will never be fully realized due to the general 

underreporting of sexual assaults. In addition, the drugs used can be difficult to 

detect (active products at low doses, chemical instability), possess amnesic 

properties, are rapidly cleared from the body (short half-life), and are difficult to 

detect when there is delay in specimen collection. To perform successful 

toxicological examinations, analysts need to obtain sufficient amount of the 

corresponding biological specimens (blood and urine as soon as possible, hair 

about 1 month after the alleged event), use sensitive methods to detect 

relevant drugs and/or metabolites, and be cautious in results interpretation. 

The Society of Forensic Toxicologists (SOFT) DFSA Committee developed a 

chart of the most prevalent drugs associated with DFSA and the associated 

recommended “maximum detection limits” when analyzing urine specimens 

[75]. To address the current challenges in dealing with DFSA cases, a series of 

reviews were published dealing with laboratory management of DFSA cases 

[ 76 ], frequency of DFSA investigation [77 ], the use of alcohol [ 78 ], 

benzodiazepines [79], GHB and its analogs [80], miscellaneous prescription 

drugs [81], over the counter drugs [82] and the “Z-drugs” (i.e., zopiclone, 

zolpidem, and Zaleplon) [83]. The analysis of benzodiazepines and their 

derivatives using their isotopically labeled analogs has also been reviewed 

[84]. 

In a review of the current understanding of alleged DFSA, several definitions of 

DFSA are provided as well as a list of the intoxicating substances which have 

so far been implicated in this type of crime [85]. Also, the effects of alcohol on 

sexual behaviour, drinking patterns, and specific quantitative research with 

very high alcohol levels in some instances, are also addressed. Reviews for 

DFSA cases [86] and in cases involving the elderly and chemical poisonings 

[87], with particular emphasis on the analysis of hair, were presented. Kintz 

commented that hair should not be considered as an alternative to blood and 

urine analyses, but as a complement. This approach may find useful 

applications, but the definition of legally defensible cutoff values would require 

much more data. In a literature research by Madea et al., the modes of action 

and toxicological means of detection of substances used as knock-out drugs 

were described. The most frequently used drug in DFSA was still alcohol 

followed by illicit drugs (cannabis, cocaine, etc.) [88]. Only in relatively few 

cases was the presence of involuntarily consumed medications and drugs of 

625

abuse found by routine toxicological analysis, with benzodiazepines most 

commonly found followed by other hypnotics. 

Alleged sexual assault cases in North Ireland from 1999 to 2005 were studied 

[89]. The estimated average blood alcohol concentration remained broadly 

similar (218 mg% in 1999 vs 217 mg% in 2005), while there was an increase in 

the detection of antidepressants, recreational drugs, benzodiazepines, and 

analgesics, some of which were also associated with alcohol consumption. 

The study, like others, failed to identify specific date-rape drugs, such as GHB 

or Rohypnol (flunitrazepam). Likewise, a study by the U.K.'s National Forensic 

Services also suggested that there was no evidence for the use of 

flunitrazepam in DFSA during its 3-year investigation [90]. Alcohol and drug 

consumption, sexual abuse history, sexual impulsivity, and risk of recidivism 

were evaluated for 104 convicts in Brazil. It was concluded that alcohol use or 

abuse was associated with the perpetration of sexual aggression. The role of 

alcohol consumption seems to be greater in sexual offenders against boys 

than against girls [91]. A study by recruiting sexual assault complainants in the 

U.S. to provide urine and hair specimens for analysis of 45 drugs estimated 

that 43% of 144 subjects, and 7% of 859 complainants, were characterized as 

victims of DFSA [92] Results of toxicological analysis of blood and urine 

specimens from 1806 female victims of alleged non-consensual sexual activity 

in Sweden were reported [93]. It was found that 772 cases had ethanol only, 

while 215 cases had ethanol together with at least one other drug. The mean 

BAC was 1.24 g/L. Amphetamine and THC were the most commonly detected 

illicit drugs in blood or urine (262 cases). Among prescription drugs, 

sedative-hypnotics such as diazepam and zopiclone were the most commonly 

found, along with SSRI antidepressants and various opiate analgesics. 

A DFSA case involving the use of acepromazine (a phenothiazine tranquilizer) 

was reported [94]. The presence of acepromazine was not detected in the 

blood but it was subsequently detected in the victim's hair, collected a month 

and a half after the incident, at a concentration of 31 pg/mg. A commonly 

available over-the-counter drug, tetrahydrozoline, was reported to induce an 

obtunded compliant victim with no memory of the period during the sexual 

assault [95]. It may be warranted to include imidazoline derivatives in the 

testing for cases of DFSA. A case of a 24-year-old girl who was sexually 

assaulted after administration of GHB, and morphine was reported [96]. 

Toxicological analysis of her hair using GC/MS with 2/3-cm segmentation of all 

626

the length (20 cm) of the hair showed a higher value of GHB, and the presence 

of morphine was found in the period associated with the alleged criminal 

activity. Current analytical methods used for screening drugs and their 

metabolites in biological samples from victims of DFSA or other vulnerable 

groups can lack sufficient sensitivity. A UPLC-MS/MS method for the detection 

of ketamine and its metabolites was developed to analyze urine specimens 

from 6 volunteers following a small oral dose of ketamine (50 mg) [97]. With 

LODs for ketamine and norketamine being 0.03 ng/mL and 0.05 ng/mL, 

respectively, detection in urine could be up to 5 and 6 days, respectively, and 

the detection could be extended up to 10 days if dehydro-norketamine is also 

analyzed. 

2.3 Workplace Drug Testing 

To combat misuse of drugs or alcohol in the workplace, the application of 

workplace drug testing has grown at a phenomenal rate around the world. The 

U.S. Department of Transportation updated its drug and alcohol testing 

procedures to make specimen validity testing mandatory [98], and also to 

authorize employers to disclose to State commercial driver licensing (CDL) 

authorities the drug and alcohol violations of employees who hold CDLs and 

operate commercial motor vehicles [99]. 

The current perspective of workplace drug testing in the U.S. was reviewed 

[100]. A number of inconsistencies were observed based on epidemiological 

data between illegal drug-use self-reported survey and drug testing results 

which might be due to the increased specificity of immunoassay screening 

tests and the critical issues of adulteration and substitution. In addition, the 

performance of rapid point-of-collection (POCT) oral fluid drug-testing devices 

was also reviewed. Most of the POCT devices can detect amphetamines and 

opiates very well, while their ability to detect cocaine varies significantly across 

devices and the detection of cannabis use is generally poor across all devices 

[100]. The Finnish legislation on workplace drug testing (2004) incorporates 

provisions related to drug use testing [101]. Their purpose is to ensure that 

drug testing is carried out by good occupational health care practice and 

laboratory quality standards, taking into account the integrity and protection of 

privacy of the persons tested as well as their other fundamental rights. In 

connection to the legislation, the guidelines to be followed by laboratories were 

described, along with the requirement for accreditation [102]. An overview of 

the New Zealand experience in drug-free workplace program was described, 

627

highlighting the mix of testing options employed, the industry sector trends, the 

categories of drugs misused, the influence of significant Employment Court 

Judgements, proposed changes to the AS/NZS 4308(2006), and current oral 

fluid research projects [103]. 

The importance of LC-MS in supporting workplace drug-testing programs was 

reviewed with particular focus on the combination of LC-MS with innovative 

instrumentation such as triple quadrupoles, ion traps, or time-of-flight mass 

spectrometers [104]. 

2.3.1 Urine 

The most common specimen for illicit drug testing is urine. Immunoassay is 

commonly used for screening because the method is simple, sensitive, rapid, 

and is reasonably cost-effective. The potential false-positive and 

false-negative results in urine drug screens that occur with immunoassays of 5 

substances (amphetamines, cannabinoids, cocaine, opiates, and 

phencyclidine) and also with alcohol, benzodiazepines, and tricyclic 

antidepressants, were discussed [105]. Other pitfalls, such as adulteration, 

substitution, and dilution of urine samples, were also included. A 

homogeneous immunoassay (EMIT-type assay) has been developed for the 

detection of zolpidem in urine, with an LOD of 5 ng/mL. The zolpidem antibody 

is highly specific and does not cross-react with zopiclone and zaleplon, and the 

accuracy of the assay was greater than 90% as demonstrated by authentic 

urine samples, with results confirmed by LC-MS/MS [106]. 

A method to analyze 40 different drugs of abuse using UPLC-MS/MS with a 

run time of 5.2 min to replace immunoassay screening for drugs of abuse in 

urine was developed. Urine samples were hydrolyzed enzymatically, diluted 

and injected with isotopically labeled internal standards; a parallel analysis with 

GC/MS showed excellent correlation results [ 107 ]. An ELISA and an 

LC-MS/MS confirmation method were developed and validated for the 

identification and quantitation of ketamine and its major metabolite 

norketamine (NK) in urine samples [108]. A cutoff value of 25 ng/mL ketamine 

in the ELISA screen was proposed. In another study, positive ion chemical 

ionization-gas chromatography-mass spectrometry (PCI-GC-MS) with an 

automatic solid-phase extraction (SPE) apparatus was used to study ketamine 

and NK in urine samples with LODs and LOQs of 25 ng/mL and 50 ng/mL, 

respectively, for both analytes [ 109 ]. Two solid phase extraction 

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(SPE)-LC-MS/MS methods for the analysis of THC-COOH in human urine 

were developed, with LOQs of 2 ng/mL [110] and 5 ng/mL [111] and no 

carryover evidenced at both 2000 ng/mL [110] and 5000 ng/mL [111]. An 

LC-MS/MS method was developed for detection of cocaine and its metabolites 

in urine at concentrations as low as 1.0 ng/mL [112]. 

Understanding urinary excretion profiles following controlled drug 

administration is important for interpretation of urine test results. The pattern 

and timeframe of excretion of MDMA and its metabolites in urine were studied, 

where placebo, 1.0 mg/kg, and 1.6 mg/kg oral MDMA doses were 

administered double-blind to healthy adult MDMA users. After 1.6 mg/kg, 

median of maximum concentrations, C(max), were 21,470 (MDMA), 2229 

(MDA), 20,793 (HMMA), and 876 ng/mL (HMA) at median time to maximum, 

T(max), of 13.9, 23.0, 9.2 and 23.3 h. Identification of 

4-hydroxy-3-methoxymethamphetamine (HMMA) as well as MDMA increased 

the ability to identify positive specimens, but required hydrolysis [113]. Huestis 

and co-workers studied the urinary excretion profile of cocaine through various 

modes of administration under controlled conditions [114,115]. Both studies 

concluded that urine ecgonine concentrations peaked later and had longer 

detection times than the other minor metabolites [114,115]. With a 50 ng/mL 

cutoff concentration, and following low doses of 10 to 45 mg cocaine by 

multiple routes, detection times extended up to 98 h [115]. The excretion 

profiles of flunitrazepam metabolites in urine after a single dose (0.5 mg or 2 

mg) were investigated using CEDIA (300 µg/L cutoff) for screening and 

quantitation using LC-MS/MS (0.5 µg/L). It was found that none of the subjects 

receiving 0.5 mg screened positive, and only 23 of 102 samples from the 

subjects given 2.0 mg were positive with CEDIA, but that 

7-aminoflunitrazepam was detected up to 120 h, and that LC-MS/MS was able 

to detect two subjects after more than 240 h administration at low dose [116]. 

In a EtG controlled administration study, the maximum EtG concentration 

normalized to creatinine after the uptake of trace amounts of 1 g and 3 g of 

ethanol was 0.32 mg/L and 1.53 mg/L, respectively, and 0.15 mg/L and 1.17 

mg/L for EtS. These peak concentrations are considered to be positive by 

many laboratories testing urine for ethanol conjugates in work-place testing 

progammes [117]. Differentiating new intake of drugs-of-abuse from residual 

drug excretion may be difficult, especially following chronic drug usage and for 

drugs with long elimination half-lives such as cannabis. In a case of young 

pregnant woman following heavy and chronic cannabis use, urinary 

11-nor-9-carboxy-delta-9-tetrahydrocannnabinol (THCCOOH) concentrations 

629

measured by LC-MS, declined from 348 to 3.9 ng/mL over a surprisingly long 

period of 12 weeks (84 days), but were supported by her THCCOOH excretion 

profile, indicating that special caution is needed in interpreting urine test results 

from heavy, chronic cannabis users [118]. To test the validity of THC and 

11-hydroxy-THC (11-OH-THC) in urine as indicators of recent cannabis use, 

urinary cannabinoid excretion in 33 chronic cannabis smokers, who resided on 

a secure research unit under 24 h continuous medical surveillance, were 

monitored. Extended urinary excretion of THC and 11-OH-THC was observed 

for at least 24 days, negating their effectiveness as biomarkers of recent 

cannabis exposure, and substantiating long terminal elimination times for 

urinary cannabinoids following chronic cannabis smoking [119]. The time 

course of THCCOOH elimination in urine was characterized in 60 cannabis 

users during 24 h monitored abstinence for up to 30 days, with urine samples 

taken for GC-MS analysis. Data were divided into three groups, 0-50, 51-150, 

and > 150 ng/mg, and results from this study showed that the number of days 

until first negative and last positive urine specimens for the three groups were 

0.6 and 4.3, 3.2 and 9.7, and 4.7 and 15.4 days, respectively [120]. 

In a retrospective study of a military testing program that had been conducted 

over a three-year period, it was found that 50% of all positive marijuana and 

cocaine urine metabolite were below median values of 65 and 968 ng/mL, 

respectively. A median drug concentration combined with the percentile or 

frequency that a particular urine drug concentration occurs, may provide useful 

information for evaluation [121]. 22 urine specimens reported by the same 

group for the presence of high concentrations of amphetamine only (28,028 to 

241,142 ng/mL) were subjected to further analysis for the presence of 

methamphetamine (MA) [ 122 ]. It has been suggested that a urine 

concentration ratio of MA to amphetamine less than 0.5%, with similar isomeric 

distribution of d-(S) and l-(R) amphetamine and MA, respectively, may not 

necessarily indicate the use of both amphetamine and MA. 

A study reported that increased storage temperatures were associated with 

increased urine pH, which might be due to degradation of nitrogenous urine 

[123]. The pH values of specimens stored at -20 o C are relatively stable, 

whereas pH results > 9 are achieved when samples are stored at room 

temperature or higher. Statistical data of consecutive urine-creatinine 

concentrations were evaluated as a laboratory marker of urine dilution [124]. 

The 2.5-97.5% percentile ranges for women were: 129-2690 mg/L, while for 

men were 204-3520 mg/L. Using cut-offs greater than the gender-specific 

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2.5% percentiles bear a high risk of misinterpretation, and gender-specific and 

creatinine-adjusted decision limits are suggested to differentiate urine 

adulteration. Two drug normalization procedures based on specific gravity and 

creatinine to compensate for excessive hydration were applied to urine 

specimens collected from three distinct groups (pain patients, heroin users, 

and marijuana/cocaine users) [ 125 ]. By applying this normalization, an 

increase in percent positives was observed, particularly for marijuana/cocaine 

users (2- to 38-fold increases). 

A literature review of the influence of poppy seed foods on opiate drug tests 

revealed that the possibility of false-positive opiate drug tests after poppy food 

ingestion is genuine and that there are no unambiguous markers available to 

differentiate poppy food ingestion from heroin or pharmaceutical morphine use 

[126]. Capsule and drink type detox agents were evaluated for their ability to 

alter the elimination of MA in rats, and no significant differences was found 

between the control and detox-dosed groups in the amounts of MA and AP 

excreted into urine or in the volume of excreted urine [127]. 

2.3.2 Alternative specimens 

The addition of accurate and reliable workplace drug testing using hair, oral 

fluid, and sweat patch specimens can complement urine-based drug testing, 

and aid in combating products intended to subvert drug testing through 

adulteration, substitution, and/or dilution. In 2004, significant revisions of the 

U.S. Department of Health and Human Services (HHS) drug testing standards 

were published defining, standardizing, and requiring specimen validity testing 

on Federal employee donor urine specimens [128]. 

2.3.2.1 Oral fluid 

Oral fluid offers multiple advantages as an alternative matrix for drug 

monitoring and therefore has an increasingly important role in DUID, treatment, 

workplace, and criminal justice programs. A review has outlined oral fluid 

testing advantages and limitations, and the progress in oral fluid testing in the 

last 5 years with respect to the collection of oral fluid and subsequent 

screening, confirmation, and results interpretation for cannabinoids, opioids, 

amphetamines, cocaine, and benzodiazepines [27]. In another review, it was 

noted that significant developments have occurred; i.e., new pharmacokinetic 

studies have been conducted, optimal cutoffs have been proposed, and the 

631

correlation between oral fluid drug concentrations and impairment have been 

examined [129]. However, reliable point-of-care drug testing is still problematic, 

especially for cannabinoids and benzodiazepines [129]. 

A database of oral fluid test results from a commercial laboratory in the U.K. 

(8679 confirmed positive results) was evaluated to study drug/metabolite 

prevalence data, concentrations, and drugs/metabolite patterns encountered 

in oral fluid [ 130 ]. Opiate positives were most frequently observed in 

legal/treatment settings, whereas cannabinoids were most frequently reported 

in the workplace. 

Although oral fluid testing is now commonplace in many monitoring programs, 

the greatest current limitation is the scarcity of controlled drug administration 

studies to guide interpretation of results. The concentrations of oxazepam and 

oxazepam glucuronide in oral fluid, blood, and serum after controlled 

administration of 15 and 30 mg oxazepam in eight subjects was determined, 

and the mean oral fluid/blood ratios were 0.05 for oxazepam and 0.004 for 

oxazepam glucuronide [131]. 

2.3.2.2 Hair 

With more demand for analyzing drugs in hair, many methods have been 

developed and validated. A commercial ELISA test for the screening of drugs 

of abuse (cannabis, amphetamines, opiates, and cocaine) in human hair was 

validated. No false negative results were observed, based on the Society of 

Hair Testing's cutoffs (0.5 ng/mg for cocaine, 0.2 ng/mg for opiates and 

amphetamines, and 0.1 ng/mg for THC) [132]. Micro-pulverized extraction has 

been used as the sample preparation procedure for analysis of MA in human 

hair analysed by either LC-MS/MS [133] or a microchip-based ELISA system 

(microELISA) [134]. The pulverizing highly disintegrated the hair components 

(2 mg of hair), thereby allowing the extraction of any drugs present for 

LC-MS/MS analysis [133]. The microELISA was run in parallel with LC/MS/MS 

and also proved to be a reliable method [134]. A rapid and cleanup-free 

microwave-assisted extraction (MAE) method was proposed for the 

simultaneous extraction of cocaine, opiates, and their associated metabolites 

from human hair samples, followed by LC-DAD detection [135]. Using 50 mg 

hair, the LOQ of the method was 0.2 ng/mg, and similar recoveries were 

obtained when compared with enzymatic or acid digestion. MAE was also 

applied for the analysis of EtG for 15 hair samples from occasional alcohol 

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users, obtaining positive results in all cases, using GC-MS with a linear range 

of 0.3-10 ng/mg [136]. 

An LC-MS/MS method was developed for the determination of ketamine (and 

its metabolite norketamine) and some amphetamines in human hair [137]. 

Using 20 mg hair, the LOD and LOQ were 0.1 and 0.5 ng/mg, respectively, for 

ketamine (K) and amphetamines. Another LC-MS/MS method for the 

simultaneous identification and quantification of amphetamines, diazepam, 

cocaine, their metabolites, and opiates from hair was developed using a single 

SPE method, with LODs between 0.02 and 0.09 ng/mg [138]. Pulverized hair 

samples were extracted with methanol, and the supernatant injected into the 

LC/MS/MS system for the simultaneous determination of opiates, cocaine, and 

their metabolites [139]. The LODs were 10 pg/mg for morphine, codeine and 

6-acetylmorphine (6-AM); and 1 pg/mg for cocaine and BE, and the method 

was successfully applied to the analysis of 79 authentic hair samples. 

A GC-MS method for determination of ketamine, amphetamines, and opiates 

in hair was developed [140]. With 25 mg hair, and following derivatization with 

heptafluorobutyric acid anhydride, the LODs were in the range of 0.03-0.08 

ng/mg, and the LOQs were 0.05-0.1 ng/mg. Using GC/NCI-MS, the 

sensitivities of amphetamine, MA, MDA, MDMA, MDEA, morphine, and 

codeine were improved from 15- to 60-fold (and 200- and 2000-fold for K and 

NK) when compared with the EI method [141]. The analysis of cocaine and 

benzoylecgonine in hair, using GC/MS after derivatization with a mixture of 

N-methyl-N-(trimethylsilyl)trifluoroacetamide with 5% chlorotrimethylsilane, 

was reported [142]. Using a sample size of 20 mg hair, the LODs and LOQs 

were 20 and 50 pg/mg for cocaine, and 15 and 50 pg/mg for benzoylecgonine, 

respectively. With the use of a commercial chiral derivatizing agent, 

trifluoroacetyl-prolyl chloride, chiral separation of amphetamines, MA, MDMA 

and MDEA in hair samples was achieved by GC/MS operated in the SIM mode, 

with LODs in the range of 0.1-0.2 ng/mg [143]. 

As with other alternative specimens, ascertaining the time of exposure and the 

extent of drug use using hair test results is difficult due to the complicated 

disposition mechanism(s) and limited research available, particularly studies in 

controlled clinical environments. Hair results from MA abusers in 2070 hair 

samples were statistically evaluated [144]. The low, medium and high ranges 

of MA were 0.5–4.2, 4.2–24.5, and 24.5–608.9 ng/mg, and those of 

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amphetamine were 0.1–0.4, 0.4–1.7, and 1.7–41.4 ng/mg, respectively. In 

addition, the amphetamine-to-MA ratios showed large variations but also a 

tendency towards smaller variations as the MA ranges increased. Segmented 

hair specimens of self-declared ecstasy abusers obtained from a double-blind 

placebo-controlled six-way crossover study, during which they received a 75 

and a 100 mg dose of racemic MDMA twice, were used to determine the 

enantioselective disposition of MDMA. With the use NCI-GC-MS, the 

(R)-enantiomer was predominant, with R/S ratios varying between 1.02 and 

2.75 and total concentrations ranging from 0.1 to 20.1 ng/mg [145]. Although 

there is a study showing no significant difference in results between natural 

and bleached hair, the results show a possible reduction in cocaine and 

benzoylecgonine concentrations for bleached hair [49,146]. In fact, there is 

another study showing that bleaching may lead to false negatives for analysis 

of EtG in hair [147]. Hence, it is possible that the bleaching or perming of hair 

may negatively affect the result of analysis. 

2.3.2.3 Sweat 

Toxicology tests obtained from 188 properly worn and unadulterated patches 

(out of 536 applied) from opiate-dependent, treatment-seeking volunteers 

showed that the level of agreement between positive sweat test results and 

positive urine results was 33% for opiates and 92% for cocaine. Also, there 

was a low acceptability of sweat patch testing by patients (only 54.3% were 

brought back attached to the skin) and that weekly sweat testing is less 

sensitive than weekly urine testing in detecting opiate use, suggesting limited 

utility of sweat patch testing in outpatient clinical settings. The excretion profile 

of methadone and other drugs (nicotine, cotinine, caffeine, and cocaine) was 

studied by analyzing sweat patches of heroin abusers by GC/MS [148]. The 

results showed that all the patches showed positive results for methadone, 

with cocaine detected in two cases, and that the parent drug was mainly 

identified rather than the metabolite(s). Sweat patches (n = 389) were collected 

from 39 pregnant volunteers and were analyzed for opiates, cocaine, and 

related metabolites using GC-MS [149]. Cocaine was present in 254 (65.3%) 

patches in concentrations ranging from 5.2 to 11,835 ng per patch, with 154 of 

them higher than the 25 ng per patch of proposed the Substance Abuse and 

Mental Health Services Administration’s (SAMHSA) cutoff. Interestingly, 

6-acetylmorphine was the most prominent opiate analyte documented in 134 

patches (34.4%), with 11.3% exceeding the cutoff of 25 ng per patch. 

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2.4 Emergence of New Drugs of Abuse 

There is increasing use of legally available “designer drugs” as recreational 

drugs. Many of these designer drugs are structurally and/or pharmacologically 

very similar to their corresponding illicit analogs. Contrary to new medications, 

which are extensively studied in animals, followed by human epidemiological 

investigations and risk assessment under controlled clinical environments, 

many designer drugs are consumed without any safety testing. 

2.4.1 Phenethylamines 

Metabolite standards are often not commercially available, particularly in the 

case of new designer drugs. As an alternative to often cumbersome chemical 

synthesis, human cytochrome P450 isoenzymes (P-450s) can be used for the 

biotechnological synthesis of drug metabolites. This concept was applied to 

the synthesis of N-(1-phenylcyclohexyl)-2-hydroxyethanamine (PCHEA), the 

common O-dealkyl metabolite of the designer drugs 

N-(1-phenylcyclohexyl)-2-methoxyethanamine (PCMEA) and 

N-(1-phenylcyclohexyl)-2-ethoxyethanamine (PCEEA) [150]. The involvement 

of P450s in the metabolism of N-(1-phenylcyclohexyl)-3-ethoxypropanamine 

(PCEPA) and N-(1-phenylcyclohexyl)-3-methoxypropanamine (PCMPA), to 

the common metabolite N-(1-phenylcyclohexyl)-3-hydroxypropanamine 

(PCHPA), was studied [151]. Among the tested isoenzymes, P450 2B6, 2C19, 

2D6, and 3A4 catalyzed PCEPA O-deethylation, and P450 2B6, 2C19, and 

2D6 catalyzed PCMPA O-demethylation. 

Another new class of designer drugs, the so-called beta-keto (bk) 

phenethylamines (aka “cathinones”) such as mephedrone 

(bk-4-methylmethamphetamine), butylone (bk-MBDB), and methylone 

(bk-MDMA) have appeared in the illicit drug markets in many countries. The 

metabolites of bk-MBDB and 

2-ethylamino-1-(3,4-methylenedioxyphenyl)propan-1-one (bk-MDEA) in 

human urine were identified using synthesized standards [152]. Based on 

GC-MS and LC-MS, N-dealkylation, demethylenation followed by 

O-methylation, and β-ketone reduction were identified as their major metabolic 

pathways. 

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Maurer et al. reported a series of studies on the metabolism of PCMPA [153], 

N-(1-phenylcyclohexyl)-propanamine (PCPR) [ 154 ], 

N-(1-phenylcyclohexyl)-2-ethoxyethanamine (PCEEA) [ 155 ] and 

N-(1-phenylcyclohexyl)-2-methoxyethanamine (PCMEA) [155], 

4-chloro-2,5-dimethoxyamphetamine [ 156 ], mephedrone, butylone, and 

methylone [157] in rats and/or humans, by analyzing urine samples using 

GC-MS after acid hydrolysis, liquid-liquid extraction, and microwave-assisted 

acetylation. The systematic analytical methods described in these reports are 

useful for screening of these drugs. 

A fatal overdose case involving the potent hallucinogenic drug 

“Bromo-Dragonfly” (1-(8-bromobenzo[1,2-b; 

4,5-b′]difuran-4-yl)-2-aminopropane) was reported [158]. Bromo-Dragonfly was 

identified in the liver and blood using UPLC–Time of Flight Mass Spectrometry 

(TOFMS), and was subsequently quantified in femoral blood (0.0047 mg/kg), 

urine (0.033 mg/kg), and vitreous humour (0.0005 mg/kg). A synthesized 

designer drug, para-methoxyethylamphetamine (PMEA) was unexpectedly 

detected in the postmortem specimens in a case in Japan [159]. Blood 

concentration of PMEA was estimated to be 12.2 µg/mL, with 

para-methoxyamphetamine, para-hydroxyethylamphetamine, and 

para-hydroxyamphetamine identified as its major metabolites. 

3-Fluoromethcathinone was identified in capsules marketed as plant fertilizer 

available from internet sources [160]. GC–MS data for the three isomers of 

fluoromethcathinone and their N-acetyl derivatives were reported, and the 

positional isomers of fluoromethcathinone were determined using Fourier 

transform infrared spectroscopy (FTIR) and 19 F nuclear magnetic resonance 

spectroscopy (NMR). 

2.4.2 Piperazines 

Reports on the use of benzylpiperazine (BZP) and 

1-(3-trifluoromethylphenyl)piperazine (TFMPP) for their stimulating, enacting, 

and "recreational" properties are increasing. In view of the abuse potential, 

BZP is banned in the U.S., Japan, Australia, Denmark, and Sweden. In 2008, 

New Zealand also controlled BZP and related substances [161]. A review of 

the information currently available on BZP suggested that further research into 

its mechanisms of action, long-term effects, and potentially addictive 

properties is needed [162]. A study to assess the abuse liability of BZP in rats 

indicated that BZP self-administration, like cocaine, is readily acquired and 

636

mediated by dopaminergic mechanisms [163]. In a national household survey 

conducted in New Zealand of the use of BZP/TFMPP showed that the mean 

quantity of BZP/TFMPP taken on an occasion of greatest use was 533 mg. It 

was concluded in the survey that females may be at greater risk of problems 

from BZP/TFMPP abuse, due to their smaller physical size, and that taking 

5-hydroxytryptophan (5-HTP) 'recovery' pills with BZP/TFMPP may actually 

increase the risk of adverse effects [164]. 

BZP intoxication can be life threatening, and it has been detected in several 

fatal cases, while the toxicity of TFMPP appears to be weaker but has a risk of 

serious psychiatric manifestations and serotoninergic syndrome [165]. Three 

cases of BZP and TFMPP intoxications were reported, with serum 

concentrations of TFMPP at 263 ±5.8 ng/mL and BZP at 46.7 ± 15.3 ng/mL 

[166]. The pharmacokinetics of BZP was measured in blood and urine samples 

taken from 7 healthy adults over 24 h following a 200 mg oral dose of BZP, 

with a peak plasma concentration at 262 ng/mL in 75 min [167]. The results of 

this study indicate that BZP may be detectable in plasma for up to 30 h 

following an oral dose. Seven urine specimens were analyzed for BZP and 

TFMPP, with concentrations ranging from 13.0 to 429.1 mg/L and 0.79 to 25.4 

mg/L, respectively [ 168 ]. A new derivative of BZP, 

4-bromo-2,5-dimethoxybenzylpiperazine, first seized in Germany, was 

reported in 2009. The structure was proven by two-dimensional NMR 

correlations and GC-MS, after synthesis of two of the possible isomers from 

commercially available starting materials [169]. 

2.4.3 Synthetic cannabinoids 

“Spice” is one of a large number of cannabis-free mixtures of various “inert” 

plant materials which have had one or more synthetic cannabinoids or 

cannabimimetics sprayed on them. Compounds such as cannabicyclohexanol, 

oleamide, JWH-018, CP 47497, JWH-073, and HU-210, have received 

intensive attention due to their cannabis-like effects when smoked, and have 

been sold on the Internet and in various specialized shops since at least 2006 

[170]. Responding to potential health concerns, Austria, Germany, France, 

Luxembourg, Poland, Lithuania, Sweden, Estonia, and a number of states in 

the U.S. have taken legal actions to ban or otherwise control these products 

and related compounds. Identification of many of the compounds used in these 

products have been reported in Japan [171,172], Germany [173,174], Spain 

[175], many European countries, and the U.S. [170]. While there are no 

637

officially published safety data concerning their effects in humans, a case was 

reported for a patient who developed physical withdrawal phenomena and a 

dependence syndrome similar to that seen in cannabis dependence, after the 

consumption of "Spice" [176]. 

2.4.4 Pyrrolidinophenones 

In the metabolism of 4'-methyl-alpha-pyrrolidinobutyrophenone (MPBP) to 

4'-(hydroxymethyl)-alpha-pyrrolidinobutyrophenone, only P450 enzymes 

CYP2D6, CYP2C19, and CYP1A2 were capable of catalyzing MPBP 

4'-hydroxylation. From the relative activity factor and experiments with human 

liver microsomes from CYP2D6 and CYP2C19 poor metabolizers, it could be 

concluded that polymorphically expressed CYP2D6 is mainly responsible for 

MPBP hydroxylation [177 ]. With the use of GC/MS, 11 metabolites of 

alpha-pyrrolidinovalerophenone (PVP) in rat urine were identified, and a 

metabolic pathway was suggested involving initial hydroxylation of the side 

chain followed by dehydrogenation to the corresponding ketone [178]. For 

studies on the quantitative involvement of P450s in the initial hydroxylation of 

1-(4-methylphenyl)-2-pyrrolidin-1-ylhexan-1-one 

(4'-methyl-alpha-pyrrolidinohexanophenone, MPHP), the metabolite 

4'-hydroxymethyl-alpha-pyrrolidinohexanophenone (HO-MPHP) was 

synthesized (for use as a reference standard) using a biotechnological 

approach [ 179 ]. The NMR and MS data of another designer drug, 

3,4-methylenedioxypyrovalerone (MDPV), which is claimed to be a stimulant 

(first seized in Germany in 2007), were described [180]. 

2.5 Survey on Trends of Drug Use 

2.5.1 Opiates 

The global epidemic of opiate use continues to spread and is an increasing 

burden, especially in developing countries. In a study of acute opiate 

overdoses in a poisoning center in Tehran, Iran over a six month period, 

showed that opium rather than heroin was most commonly used by regular 

users, with benzodiazepines, antidepressants, and alcohol being the most 

common agents co-consumed with opiates [181]. The pattern of poisoning in 

the elderly population in Tehran also found that opioids and opiate products 

accounted for 54.02% of the non-pharmaceuticals found, and that the most 

frequently involved drug groups were benzodiazepines, antidepressants, and 

638

analgesics [182]. An epidemic of fatal accidental poisonings due to the highly 

potent opioid-mimicking designer drug 3-methylfentanyl (TMF) was revealed 

among Estonian drug users, with 46 and 71 cases found in 2005 and 2006, 

respectively. The mean combined concentration of TMF stereoisomers among 

pure TMF cases (1.9 µg/L) was more than ten times lower than the mean 

fentanyl concentration in fentanyl-related fatalities [183]. 

2.5.2 Amphetamines 

The abuse of amphetamines is a major threat worldwide, and various studies 

on the epidemic in Australia, the U.S., and southeast Asia were reported. In a 

study of MA-related deaths in Australia, injection was found to be the major 

route of administration (89%), with other drugs detected including 

benzodiazepines (41%) and morphine (36%) [184]. The median blood MA 

concentration was 0.2 mg/L (range 0.02-15.0 mg/L). A study of abuse of MA 

and amphetamine in North America indicated the annual prevalence for 

'speed' use in Canada being 0.8% in 2004, 0.3% for amphetamine and 0.1% 

for MA in Mexico in 2002, and 1.4% for 'stimulants' in the United States in 2006 

[ 185 ]. The type of stimulants has shifted from non-medical use of 

pharmaceutical amphetamine to use of MA powder and then 'ice'. Data 

collected in southeast and east Asia indicated that they have experienced a 

MA epidemic in the past decade, which began around 1997 and peaked in 

2000-2001 [186]. MA is typically smoked or ingested, but injection of the drug 

is also apparent. The bone mineral density in the lumbar spine measured 

among 46 hospitalized male MA abusers in Korea indicated that there was a 

considerable loss of bone mineral among them [187]. 

In a study of 169 forensic autopsy cases detected with amphetamines in 

Australia, 9 cases with either cerebral hemorrhage or serotonin syndrome 

were caused by the interaction of MDMA and moclobemide, and 19 cases with 

heart disease were associated with long-term use of amphetamines [188]. 

Another study in Australia for MDMA related deaths based on the National 

Coronial Information System showed that MDMA deaths were predominantly 

due to drug toxicity (82%), with MDMA being the sole drug causing death in 

23% of cases, and combined drug toxicity in 59% of cases [189]. A survey of 

the laboratory urinalysis data of ecstasy users in the U.S. in 2005-2007 

showed that a majority (84.3%) of the specimens contained multiple drugs 

and/or metabolites in addition to MDMA and MDA, with the median number of 

drugs/metabolites reported being 5. The most commonly identified drug 

639

groups were in the following order: Cannabis (THCCOOH); amphetamine/MA; 

benzoylecgonine; diazepam-related; and opiates and alprazolam [190]. 59 

fatalities that tested positive for MDMA during the period of 2001 to 2008 in 

Taiwan were studied [191]. Among the fatalities, ketamine was found in 28 

cases, indicating the popularity of this drug in Taiwan. The cause of death was 

mainly acute intoxication, with a mean MDMA concentration of 2.75 µg/mL, 

while the manner of death was predominately accidental. 

2.5.3 GHB 

Several studies of GHB use were conducted in the U.K. [192,193,194]. 189 

individuals that reported at least one lifetime use of GHB in the U.K. completed 

an online questionnaire, which indicated that the primary use of GHB was for 

recreation [192]. In another study on 173 individuals with recreational drug 

toxicity and requiring "medical" assistance in a large urban nightclub from 

September 2007 to January 2008, the results showed that the majority of them 

were not first-time recreational drug users, and that they used a variety of 

recreational drugs, particularly GHB, gamma-butyrolactone (GBL), and 

ketamine [193]. Although the use of GHB is controlled in many countries, its 

precursors GBL and 1,4-butanediol (1,4-BD) remain legally available despite 

having similar clinical effects (GBL is now controlled in U.K., Canada, Australia, 

and Sweden). A self-reported study found that GHB ingestion was much more 

common than GBL ingestion, whereas GBL was more commonly found in the 

seized samples, suggesting that GBL use may be more common than 

previously thought [194]. 

A National Drug Strategy Household Survey in Australia showed that 0.5% of 

Australians aged 14 years or older reported ever using GHB, and 0.1% 

reported recent use, with the prevalence of use being highest amongst those 

aged 20-29 years [195]. 

2.5.4 Ketamine 

In a national survey in Taiwan from 2004 to 2006, a decline was noted in the 

prevalence and incidence rate of ecstasy, a leading illegal drug used by 

school-attending adolescents in Taiwan since the early 2000s. The emerging 

ketamine use trend, being the first commonly used illegal drug in the middle 

school-aged students starting in 2006, may warrant more attention [196]. Data 

analyzed from the 2004 National Drug Strategy Household Survey in Australia 

640

showed that lifetime use of ketamine was reported by 1% of Australians aged 

14 years or older, with 0.3% reporting recent use. Again, the prevalence of 

ketamine use was highest amongst those aged 20-29 years [195]. An interview 

was conducted for 90 ketamine users in the U.K. showed that frequent 

ketamine users expressed more concerns than other groups about long-term 

effects on physical health problems, especially “K-cramps” and cystitis, 

whereas ex-users were more concerned about mental health problems [197]. 

An assessment of 360 club-drug users and 303 non-drug users in Hong Kong 

showed that the most frequently used drugs were ketamine, ecstasy, and 

cannabis, with the majority using two or more drugs on any one occasion, and 

that club-drug users were more rash-impulsive, reward-driven, and less 

punishment-sensitive, than non-drug users [198]. 

2.5.5 Other Drugs 

A 10-year retrospective review of 216 poisoning death cases in Hubei, China 

showed that the most common classes of substances involved were 

rodenticide (19.7%), insecticide and herbicide (17.9%), carbon monoxide 

(16.5%), drugs (13.8%) and alcohols (12.4%) [199]. A retrospective evaluation 

of drugs detected in a pediatric postmortem population between the years 

1998 and 2002 (n = 730) in the U.S. state of Ohio reported that 

resuscitative/treatment drugs were detected most frequently (56% of positive 

results), followed by illicit drugs (26%), ethanol (11%), carbon monoxide (8%), 

and antihistamines (6%) [200]. The authors suggested that illicit drugs and 

ethanol be targeted for testing, especially when limited specimens are 

available for analysis. Causes of accidental drug deaths occurring in 2002 in 

the U.S. state of Georgia revealed that cocaine was the most prevalent drug 

(40%), followed by mixed drug intoxication (37%), opioids (10%), ethanol (7%), 

and prescription medications (non-opioid) (5%). This study also supported of 

the lack of correlation between serum drug levels and the mechanism of drug 

toxicity in cocaine-related deaths [201]. 

Serotonin-active drugs are detected relatively frequently in Victorian deaths in 

Australia. A study of 1123 fatalities with serotonin-active drugs (tramadol, 

venlafaxine, fluoxetine, sertraline, citalopram, paroxetine, and MDMA) 

detected over a 7-year period was conducted [202]. Of the 28 death cases 

where the circumstances suggested the involvement of serotonin toxicity, 

tramadol was frequently detected, in combination with serotonergic 

antidepressants. MDMA was also detected relatively commonly and was 

641

associated with moclobemide in 4 cases of confirmed serotonin toxicity. 

PubMed/Medline search using terms dextromethorphan (DM) and abuse 

through July, 2008 indicated that DM is often abused to seek dissociative 

effects similar to ketamine and phencyclidine, and that its abuse potential 

should be recognized in view of its low cost and ready accessibility as an 

over-the-counter drug [203]. 

Patients with Parkinson's disease (PD) frequently experience pain. A study 

with 11,466 PD patients showed that they significantly received more 

prescriptions of analgesics than the general population (82% versus 77%), and 

that they were more exposed than the general population and diabetics to 

opiates, acetaminophen, and adjuvant analgesics chronic use [204]. 

A study of 24 multiple suicidal/accidental drug intoxication cases in the U.S. 

found that opiates, antidepressants, benzodiazepines, ethanol, and cocaine 

were equally likely to be present in both suicide and accidents, and that the 

mean number of drugs found in suicides was 4, compared to 2 in accidents 

[205]. Travel-related poisonings (i.e., incidents associated with bus, taxi, train, 

and air travel, or at local markets) is an emerging social and public health 

emergency in Bangladesh. A prospective clinical study and retrospective 

analysis of hospital records suggested that benzodiazepines such as 

lorazepam, diazepam, and nitrazepam are commonly used to commit these 

crimes [206]. The blood concentrations of clonazepam and its metabolite 

7-aminoclonazepam in toxicology cases in Denmark were reported [207]. The 

median concentrations of clonazepam plus 7-aminoclonazepam were 0.067 

mg/kg for traffic cases, 0.071 mg/kg for criminal cases (offenders), 0.030 

mg/kg for criminal cases (victims), and 0.115 mg/kg for postmortem cases. 

A retrospective study of the prevalence of drugs found in carbon monoxide 

related deaths in the U.S. state of Ohio showed that the manners of death 

were mostly accidental (69%), followed by suicide (25%), and then homicide 

(6%). In addition, 28% of the deaths had ethanol detected, while 41% had 

therapeutic and/or abused drugs detected; antidepressants and antihistamines 

were commonly detected in suicides, and pain medications and antihistamines 

in accidents [208]. While it is known that diesel fuel combustion engines 

produce much lower concentrations of CO than gasoline engines, a case of 

CO poisoning of a truck driver (which was initially referred for autopsy as a 

presumed natural death) was reported [209]. Toxicology results showed lethal 

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lood carboxyhemoglobin, and the cause of death was re-certified as CO 

intoxication. A 10-year retrospective review conducted on all non-fire related 

CO deaths autopsied (n = 94) in the U.S. from 1994 to 2003, along with an 

extensive literature review, indicated no previously reported cases of fatal CO 

poisoning attributed to diesel fuel exhaust. 

2.6 Quality Assurance and Accreditation 

The implementation of quality management systems and accreditation of 

laboratories by regional or national accreditation bodies are intended to ensure 

that the laboratories are capable of providing effective and “fit for purpose” 

analytical service to the end users. The current status of accreditation amongst 

laboratories offering drug testing of hair was assessed. Although the vast 

majority of laboratories follow current guidelines (83%), only nine laboratories 

were accredited to ISO/IEC 17025 standards for hair testing; however, a 

significant number of laboratories reported that they were in the process of 

developing quality assurance protocols with a view to accrediting their 

methods within 2–3 years [210]. Quality assurance in toxicological examination 

in DUID cases requires efforts from professionals of different disciplines. In the 

quality assurance program of Switzerland for laboratories performing 

toxicological road traffic analyses, a multidisciplinary working group, the Road 

Traffic Commission (RTC), was established to organize external quality control, 

to interpret the results of these controls, to perform audits in the laboratories 

and to report all results to Swiss Federal Road Office [211]. The program 

required that all tested drugs and psychoactive substances listed in a federal 

instruction as 'zero tolerance substances' (THC, morphine, cocaine, 

amphetamine, MA, MDMA, and MDEA) and their metabolites to be tested 

once a year, and all other substances (benzodiazepines, zolpidem, 

phenobarbital, etc.) be tested periodically. External audits are performed 

triennially, but accredited laboratories can combine the audit with the approval 

of the Swiss Accreditation Service. 

Participation in external quality assurance or proficiency testing (PT) schemes 

is an important part of quality management. The Federal Aviation 

Administration of the U.S. reported the second 7 years of their postmortem 

toxicology proficiency test program [212]. Twenty-eight survey samples (12 

urine, 9 blood, and 7 tissue homogenate) with/without alcohols/volatiles, drugs, 

and/or putrefactive amine(s) were submitted, with an average of 25 

participants reporting their results. Analytes in the survey samples were 

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correctly identified and quantitated by a large number of participants, but some 

false positives of concern were reported. 

Three exercises were offered by an external proficiency testing program 

(HAIRVEQ) in 2006 to evaluate 32 laboratories' performance in analyzing the 

same hair samples containing opiates, cocaine, and methadone, after carrying 

out some specific educational interventions [213]. It was found that with the 

standard operating procedures sent together with the test hair samples, a 

reduction in the number of incorrect results was observed compared with 

previous exercises in 2004 and 2005. An external quality assessment scheme 

was reported that assessed the reliability of analytical laboratories when 

analyzing drugs of abuse in oral fluid (ORALVEQ). Results were evaluated 

from a qualitative and quantitative point of view. Eighteen false-positive and 3 

false-negative results were reported by 8 (from a total of 21) laboratories. In 

general, the participating laboratories had a satisfactory performance [214]. 

Reference materials are a critical tool for successful quality assurance and 

accreditation. Although the importance of analysis of drug in hair has attracted 

much attention in the past few years, the need for a reference material (RM) for 

method development and validation, internal quality control, or proficiency 

tests has rapidly increased. RMs of drug-free hair spiked with MA and its main 

metabolite, amphetamine (AP) [215], and authentic hair samples from MA 

abusers [216], were prepared. As a result, a spiked hair RM containing MA and 

AP was prepared at levels of 4.86±0.69 ng/mg and 4.63±0.44 ng/mg, 

respectively, and for the authentic hair RM, MA was found with concentration 

of 7.64±1.24 ng/mg and AP at 0.54±0.07 ng/mg. Certified solution standards 

(widely used in toxicology laboratories) are often purchased from vendors who 

present concentration and uncertainty differently on their Certificates of 

Analysis. Gates et al. described the impact of uncertainty associated with the 

neat material purity, mass measurement (weighing techniques), and solvent 

addition (solution density) on the overall uncertainty of the certified 

concentration, along with uncertainty calculations [217]. The measurement 

uncertainties (MUs) were estimated for the determination of MA and 

amphetamine near their cutoff concentrations in human hair [218]. With the 

use of 1% HCl in methanol, followed by derivatization and quantification using 

GC-MS, the concentrations of MA and AP in hair with their expanded 

uncertainties were 0.66+/-0.05 and 1.01+/-0.06 ng/mg, respectively. The 

method precision and the weight of the hair sample gave the largest 

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contributions to the overall combined uncertainties of MA and AP. 

Appropriate specimen selection and collection is important for a successful 

systematic toxicological analysis (STA). The Committee of STA published 

guidelines to address the issue of sampling such as specimen types, amounts 

that should be collected and submitted to laboratories expected to perform 

STA, and criteria for ensuring quality assurance in sample collection [219]. 

3. Advances in Toxicological Analysis 

3.1 Instrumental techniques 

The first step of toxicological analysis is the identification of unknown drugs 

and poisons in body fluids. A general unknown screening procedure is 

necessary to enable simultaneous detection of as many intoxicants as possible. 

Modern analytical and instrumental methods that were used in forensic 

toxicology for both qualitative and quantitative analysis of drugs have been 

reviewed [220]. The review focused on description of the theory, inherent 

strengths, and limitations of various analytical instrumentation, including 

GC-MSs, LC-MSs, mass spectrometers, and capillary electrophoresis 

instruments, commonly used in forensic toxicology. 

3.1.1 GC-MS 

The applicability of a freeware deconvolution software AMDIS (Automated 

Mass Spectral Deconvolution and Identification System) for GC-MS-based 

systematic toxicological analysis (STA) in urine was evaluated [221]. It is 

concluded that with the use of optimized settings, the AMDIS-based data 

analysis was comparable or even superior to manual evaluation by an 

experienced toxicologist, and reduced by half the overall analysis time. 

Nevertheless, AMDIS can identify only targets present in the user-defined 

target library, and may therefore not indicate unknown compounds that might 

be relevant in clinical and forensic toxicology. A comprehensive GC-MS 

screening method was developed for the detection of over 150 prohibited 

substances on the list of the World Anti-Doping Agency, including anabolic 

androgenic steroids, stimulants, and narcotic agents [222]. The analytes were 

extracted from urine by LLE using diethylether and dibutyl ether at pH 9.5 and 

14, respectively, followed by derivatization using a mixture of 

N-methyl-N-trimethylsilyltrifluoroacetamide, NH4I, and ethanethiol, prior to 

645

GC-MS analysis. 

Different workup procedures, i.e., cleavage of conjugates by acid hydrolysis 

(Hy) or enzymatic hydrolysis (Gluc) followed by liquid-liquid extraction (LLE) or 

SPE, and derivatization, e.g., acetylation (Ac), in extractive methylation (ExMe), 

for systematic toxicological screening in urine by GC-MS, were evaluated [223]. 

In conclusion, Hy-LLE-Ac remains the method of choice for STA in clinical 

toxicology owing to its wide analyte spectrum and short workup time, while 

Gluc-ExMe is an ideal complementary method when acidic drugs need to be 

covered. 

3.1.2 LC-MS 

The multi-analyte single-stage and tandem liquid chromatography-mass 

spectrometry procedures using different mass analyzers (quadrupole, ion 

trap, time-of-flight) for screening, identification, and/or quantification of drugs, 

poisons, and/or their metabolites in blood, plasma, serum, or urine published 

after 2004, was reviewed [224]. LC-MS/MS has been used as an alternative 

method for screening and quantification of drugs in biological samples, 

rodenticides [225], pesticides [226], anti-malarial drugs [227], doping controls 

[228] and abused drugs [229,230,231,232,233]. 

Apart from being a very useful and sensitive confirmatory tool, promising 

progress has been made in the application of LC-MS(MS) in general unknown 

screening (GUS). Several groups have created libraries suitable for screening 

of drugs/poisons in biological specimens [234,235,236]. With a hybrid linear 

ion-trap LC-MS/MS system, using 3 different collision energies during each 

scan, Weinmann and coworkers have created a library containing 1253 

compounds relevant in clinical and forensic toxicology by injecting pure 

solutions of drugs and toxic compounds [235]. More recently, they also 

reported a multi-target screening method to allow the simultaneous 

identification of 700 drugs and metabolites in biological fluids. Retention times 

were determined and transitions for each compound were selected by a 

"scheduled" survey MRM scan [237]. With the help of the software for routine 

screening and quantitation to allow automated acquisition and library 

searching, the time for evaluation and interpretation of the results could be 

drastically reduced. Using a similar instrument, Liu et al. has also created a 

library of LC/ESI-MS/MS for 800 drugs and toxic compounds, based on 

spectra and retention time of the analytes with increased collision energy from 

646

0.3 to 2.0 V, to produce an optimal set of fragment-rich MS/MS spectra for 

application of GUS in postmortem blood, urine, and hair specimens [234]. 

Identification of metabolites in addition to parent drugs are also helpful in the 

GUS, and five cases were used to illustrate the utility of screening of xenobiotic 

metabolites in routine analysis of forensic samples by employing a previously 

published LC-MS/MS GUS procedure [236]. 

An UPLC-TOFMS method was developed for the routine analysis of urine 

samples [238]. By comparison of acquired data to libraries containing more 

than 300 common drugs and metabolites, identification was based on a 

combination of retention time, exact mass and fragmentation patterns. The use 

of TOFMS techniques for screening of drugs in serum [ 239 ], urine 

[238,239,240,241] and hair [242,243] have been reported. 

Apart from high sensitivity and selectivity, there is also a need of 

high-throughput methods to cope with the increasing number of samples 

typically submitted to toxicology laboratories. UPLC with sub-2 µm particles 

can be operated at higher pressures, resulting in an increase in resolution, 

speed and sensitivity comparing with conventional LC [244]. Because the triple 

quadrupole MS/MS technique is limited by the cycle time when dealing with a 

large amounts of compounds, TOFMS coupled to UPLC is more frequently 

adopted in the screening of unknown analytes [240]. For example, a 

UPLC-QTOFMS method was developed for fast analysis of 103 doping agents 

(such as stimulants, diuretics, narcotics, and anti-estrogens) in urine based on 

retention time and mass accuracy, including an automated tool for peak 

picking, with a runtime of 9 min for the screening [240] and 7 min for 

confirmation with an addition use of SPE [245]. UPLC-TOFMS can be applied 

not only in screening of unknown analystes but also for quantitative analysis. 

Quantitative analysis using UPLC-TOFMS of 52 common pharmaceuticals and 

abused drugs in hair was reported [242]. But the TOF method was found to be 

less suitable for quantification of amphetamines, as high background noise 

may be observed because of its low molecular mass, leading to a lack of 

linearity and poor sensitivity [242]. Identification of drugs metabolites is 

sometimes helpful for the identification of their parent compounds, and 

software that could predict the metabolites and mass fragmentation pattern of 

quetiapine (as analysed by LC-TOFMS) was developed and was found to be 

able to assign 13 metabolites in urine samples [246]. 

647

Reverse phase liquid chromatography (RPLC) has been widely used in 

bioanalysis, but very polar compounds may be weakly retained. With the use 

of a hydrophilic stationary phase and low-aqueous/ high-organic mobile phase, 

hydrophilic interaction chromatography (HILIC) has been demonstrated to be a 

powerful technique for the retention of polar analytes [247]. HILIC also proved 

to improve MS sensitivity, allow higher flow rates, and reduce matrix effects 

[248]. Because of its advantages over RPLC, there is an increased interest in 

using HILIC in bio-analytical applications for drugs and their metabolites, which 

are often polar in nature [244,249]. For example, cocaine and its metabolites, 

especially ecgonine methyl ester, in hair samples [250], omeprazole and 

lansoprazole in plasma [251], metoclopramide in plasma [252], estrogen 

conjugates in urine [253], and acetylcholine, choline, and butyrobetaine in liver 

tissues [254]. Analysis of polar analytes, such as the endogenous metabolite 

trigonelline and the polar antibiotics amoxicillin in urine, using HILIC coupled to 

NMR and MS, was also reported [255]. Analysis of EtG, could also be 

performed by HILIC-MS/MS instead of GC; the latter requires derivatisation 

[256]. 

Although HILIC gives good separations of polar compounds, neutral and 

non-polar analytes generally show little retention [257]. To broaden the elution 

window, pharmaceutical compounds were analysed using serial coupling of 

reversed-phase and hydrophilic interaction liquid chromatography 

(RPLC/HILIC) [258]. In addition, the use of low-aqueous/ high-organic mobile 

phases in HILIC makes it possible for the eluent from SPE or liquid-liquid 

extraction (LLE) to be directly injected to the HILIC column without 

reconstitution, enabling high throughput and automated extractions 

[247,251,255]. 

3.1.3 Capillary electrophoresis 

The current application of capillary electrophoresis (CE) in forensic toxicology 

has been reviewed with special focus on those areas where this technique has 

shown real advantages over chromatographic methods, such as chiral drugs, 

proteins, and alcohol biomarkers analyses [259]. 

The utilization of dynamic doubly coated capillaries for a fast separation of 

methadone and its two major metabolites, 

2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) and 

2-ethyl-5-methyl-3,3-diphenyl-1-pyrroline (EMDP), was investigated. Complete 

648

achiral separation of methadone, EDDP and EMDP was achieved, using a 

capillary coated with a polycation of poly(diallyldimethylammonium chloride) 

and a polyanion of dextran sulfate, with migration times of approximately 4 min 

[260]. To further separate enantiomers of methadone and EDDP, another CE 

method was developed using 0.2% highly sulphated gamma-cyclodextrins as 

chiral selectors, giving LODs of 2.3-2.4 ng/mL with a 5 min runtime [261]. The 

prevalence of elevated levels of CDT, a marker of chronic alcohol abuse in 

serum of DUI drivers, was studied by CE. With a cut-off set at 2.00% (CDT 

index), 24 out of 40 drunken drivers were identified as chronic alcohol abusers 

[262]. 

Due to its selectivity and sensitivity, CE coupled to MS (CE-MS) has evolved 

as a useful analytical tool for determining drugs and metabolites in biological 

samples. Non-aqueous CE online coupled with a mass spectrometer for the 

analysis of phosphatidylethanol (Peth), another biomarker of chronic alcohol 

abuse in blood, was developed. With the use of phosphatidylbutanol as an 

internal standard, the estimated LOD was 0.1 µM, while LOQ was 0.4 µM [263]. 

Determinations of illicit drugs and/or metabolites (opiates, cocaine, 

amphetamines, and methadone) in blood [264] and in hair [265] by CE-MS, 

were accomplished. LODs in blood were in the range of 2-10 ng/mL, while in 

hair were 0.1 ng/mg. With the use of protein precipitation (PP) prior to a 

hydrodynamic injection (HD) or liquid-liquid extraction (LLE) with electrokinetic 

injection (EK), CE-ESI-MS methods for analysis of ecstasy and methadone in 

plasma, were developed. Methods were linear over the concentration range of 

0.50-175 ng/mL and 0.25-5 µg/mL for LLE-EK and PP-HD procedures, 

respectively [266]. 

3.2 Extraction Techniques 

3.2.1 Liquid-phase microextraction (LPME) 

Liquid-liquid extraction (LLE) was one of the conventional sample preparation 

techniques extensively used for biological sample analysis, particularly for 

post-mortem toxicology. This method, however, is relatively time-consuming 

and produces relatively large amount of organic solvent waste. Liquid-phase 

microextraction (LPME), which requires much less organic solvent in the 

pretreatment procedure, was recently developed [267,268,269]. There are 

several types of LPME, including single-drop microextraction (SDME) [270], 

dispersive liquid-liquid microextraction (DLLME) [ 271 ], and hollow-fiber 

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microextraction (HF-LPME) [272]. 

Hollow fibre liquid-phase microextraction (HF-LPME) has an improved stability 

and reliability over the SDME method, and may become the most widely used 

LPME method [244]. It is an efficient technique for sample clean-up, with a 

good extraction efficiency [273]. Its main disadvantage is that polar analytes 

are difficult to extract, and the pH of the sample solution must be adjusted in 

order to have total ionization of the analytes [267]. Recent studies using 

HF-LPME included the determination of some antidepressant drugs 

(imipramine, amitriptyline, and sertraline) in urine and plasma by HPLC [274], 

progesterone in human serum by GC-MS [275], β-agonists and β-blockers 

(clenbuterol, metoprolol, and propranolol) in urine, with derivatization by 

N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) followed by GC-MS 

[276], xenobiotics (DNP, DNOC, Dinoseb, and Dinoterb) in human plasma 

using LC/ESI-MS [ 277 ], an anti-diabetic drug (pioglitazone) [ 278 ], and 

antidepressants (amitriptyline, imipramine, and sertraline) [279] in biological 

fluids by HPLC. An automated method was also developed for the 

determination of flunitrazepam in plasma and urine samples using GC-MS/MS 

[280]. 

3.2.2 Electrokinetic membrane extraction (EME) 

In HF-LPME the driving force for the extraction of analytes is the passive 

diffusion, and hence the extraction times typically range from 15 - 60 min 

[267,281]. In electrokinetic membrane extraction, two electrodes, one in the 

sample solution and the other in the acceptor solution, are placed such that 

electrical field is the driving force for the extraction process; therefore, the 

extraction time can be shortened to approximately 5 – 10 min [281,282]. 

Recent studies were reported using EME on basic drugs and acidic drugs 

[283], basic drugs (pethidine, nortriptyline, tramadol, methadone, haloperidol, 

and loperamide) from untreated human plasma and whole blood, with 

recoveries at range 19 – 51% [284]. Basic drugs extraction using EME at low 

voltage (9V) (obtainable by common batteries) was demonstrated by Kjeisen 

et al. [285]. 

3.2.3 Solid-phase extractions (SPE) 

Solid-phase extractions (SPE) are becoming a more popular extraction 

method over the traditional LLE due to its lower solvent consumption, higher 

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extraction efficiency for certain drugs, and ease of automation for on-line 

coupling of SPE to chromatographic systems [286,287]. Pre-treated samples 

were injected into the LC system with two extraction columns. A comparison of 

the LODs of LLE and SPE for alkaline drugs was conducted, and most of the 

drugs had lower LODs using SPE versus LLE [288]. In addition, several 

methods have been reported to use online coupling of SPE to LC-MS/MS for 

the analysis of basic drugs of abuse in human serum [289], 21 therapeutic and 

21 drugs of abuse in urine [290], tricyclic antidepressant drugs (amitriptyline, 

desipramine, imipramine, and nortriptyline) in serum [ 291 ], 19 drugs of 

different classes in equine plasma [292], and amphetamines in blood and urine 

[293]. 

3.2.4 Microextraction in packed sorbent (MEPS) 

Although SPE has many advantages over LLE, it normally has a higher cost 

because the SPE cartridges are typically disposed of after each use. To 

address this, the microextraction in packed sorbent (MEPS) technique was 

developed, where the packed syringe can be used more than 100 times (and 

also allowing the use of smaller sample sizes, as small as 10 µL) [294]. The 

use of MEPS in sample preparation was reported for cotinine [295] and 

metabolites of monoterpenes [296] in urine by GC-MS, cocaine and its 

metabolites in urine by real-time source coupled to TOFMS [297], anti-cancer 

drugs such as cyclophosphamide [298] and busulphan [299] in plasma, and 

β-blockers such as acebutolol and metoprolol in plasma and urine by 

LC-MS(MS) [300], anesthetic drugs in plasma [301], and fluoroquinolones in 

urine by CE-MS [302]. Determination of dopamine and serotonin with MEPS 

connected online to LC-MS/MS was reported to be a facile, more rapid, and 

lower cost method [303]. 

3.2.5 Solid-phase microextraction (SPME) 

SPME has several advantages such as ease of automation, good linearity, and 

high sensitivity without the need of solvents. It can also be utilized for direct 

measurement of biological samples such as whole blood or plasma [304]. But 

longer extraction times are required for analytes with low volatility in complex 

matrices, and the capacity of SPME fiber is limited [268]. 

Automation of SPME on a 96-multi-well plate format improves the speed and 

throughput of the method to be at least comparable to automated LLE, 

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producing high-throughput SPME-LC methods [305,306,307]. While sorbents 

in conventional 96-well systems are normally located in a polypropylene 

pipette tip, a new sorbent with a monolithic plug was recently developed with a 

high binding capacity and low back-pressure properties [308,309]. 96 samples 

could be treated in 2-4 min with a good accuracy and precision. Monolithic 

packed 96-tips have been used to extract anti-cancer drugs (busulphan, 

cyclophosphamide, and roscovitine) [ 310 ], β-blocker drugs (pindolol and 

metoprolol) [311], local anesthetics (ropivacaine and bupivacaine) [312], and 

antidepressants (fluoxetine, setraline, paroxetine, citalopram, duloxetine, and 

mirtazapine) [313] in plasma. 

Other new coatings for SPME devices include sol-gel technology and 

polypyrrole (PPY) coatings, which can be electrochemically discharged and 

recharged when deposited on a metal wire [304, 314 ]. Among the two, 

applications of PPY coated-fibre SPME is more focused on analysis of drugs. 

The screening of methamphetamines in serum by SPME using 

dodecylsulfate-doped PPY film has been reported [315]. In addition to a PPY 

coating, other types of coatings for SPME of drugs in plasma have also been 

studied [316,317,318]. 

3.3 Analysis of Specific Drugs 

3.3.1 Markers for ethanol consumption 

The diagnosis of alcoholism is a topical subject of discussion; in fact, many 

studies have been published on the determination of biochemical markers of 

alcohol abuse. EtG, EtS, 5-hydroxytryptophol-glucuronide (GTOL), 

5-hydroxyindole-3-acetic acid (5-HIAA), and FAEE are all suggested as 

biomarkers for recent alcohol ingestion, with longer detection times than 

measurement of ethanol itself. The determination of EtG and/or EtS are 

commonly performed by LC-MS/MS in blood/serum [319,320,321,322], urine 

[319, 323 , 324 ], vitreous humor [321], oral fluid [319, 325 ], and hair 

[326,327,328], but methods using GC-MS are also reported [136,329]. A direct 

UPLC-MS/MS method to determine urinary GTOL and 5-HIAA was developed 

[330]. The measuring ranges were 6.7-10,000 nmol/L for GTOL and 0.07-100 

µmol/L for 5-HIAA. A direct injection UPLC-MS/MS method for measuring the 

urinary ratio of GTOL and 5-HIAA was compared with that of an ELISA assay 

for direct determination of GTOL in urine [331]. 

652

The testing of the FAEE (such as ethyl myristate, ethyl palmitate, ethyl oleate, 

and ethyl stearate) in hair as a biomarker of excessive alcohol use, was 

documented. Using a cutoff level of 0.5 ng/mg, 324 hair samples from parents 

of at-risk children analyzed by GC-MS were found with 33.3% positive rate of 

excessive drinking [332]. Of 26 FAEE hair tests in the study where the women 

donors were reported to be pregnant, 19% were positive for excessive drinking. 

In another study, FAEE and EtG concentrations in hair were determined using 

SPME-GC-MS and GC-NCI-MS, respectively [333]. The LODs were both in 

the range of 1 to 10 pg/mg. From their results, it is suggested that strict 

abstinence is excluded or improbable when concentration of FAEE > 0.2 

ng/mg or EtG > 7 pg/mg, whereas FAEE above 0.5 ng/mg and EtG above 25 

pg/mg suggested probable alcohol abuse. 

3.3.2 Small molecules 

Carbon monoxide (CO), cyanide (CN), and sulfide are well-known small and 

highly toxic molecules. Analysis for these molecules should be considered 

whenever poisoning due to these compounds is suspected. Luchini et al. has 

modified the method for determination of the carboxyhemoglobin 

concentration using spectrophotometric measurement at 420 nm and 432 nm. 

The method requires determination of the constant factors F1, F2, and F3 

every time an analysis is performed, in order to increase the accuracy and 

precision of the test [334]. A method based on solid-supported liquid-liquid 

extraction (SLE) and GC-MS, with two-step derivatization for determination of 

CN in plasma and urine, was reported [335]. The LODs of the final analyte, 

heptafluorobutyric acid-alpha-cyanobenzyl ester, were 0.04 and 0.01 µg/mL 

for plasma and urine, respectively. A method for the determination of 

thiosulfate in human urine was developed based on precolumn derivatization 

with 2-chloro-1-methylquinolinium tetrafluoroborate, followed by 

reversed-phase LC separation and ultraviolet detection of 

1-methyl-2-thioquinolone at 375 nm [336]. The LOD and LOQ were 0.3 and 0.5 

µmol/L, respectively with linear calibration range of 0.5-50 µmol/L. 

3.3.3 Drugs of abuse 

LC-MS/MS methods for simultaneous quantification of THC, 11-OH-THC, and 

THC-COOH in blood were reported either using LLE with hexane:ethyl acetate 

(90:10, v/v) [337], or by protein precipitation followed by SPE [338] of the 3 

analytes. LOQs for all analytes in the methods were in the range of 0.5-2 

653

ng/mL. The combination of the automated online SPE with LC-MS/MS can 

significantly reduce time for sample preparation requirements. With the use of 

tandem enzyme-alkaline hydrolysis, the total content of THC, 11-OH-THC, and 

THC-COOH in urine could be determined by protein precipitation followed by 

BSTFA deriviatization prior to GC-MS analysis. The calibration curves were 

linear from 2.5 to 300 ng/mL for all analytes [339]. A two-dimensional GC-MS 

(2D-GCMS) assay provides a new means of simultaneously quantifying 

cannabidiol, THC, and metabolite biomarkers. Analytes were extracted from 1 

mL of plasma by SPE, derivatized with N,O-bis(trimethylsilyl) 

trifluoroacetamide with 1% trimethylchlorosilane, and analyzed by 2D-GCMS 

with cryofocusing. The lowest points of calibration for all analytes were either 

0.25 or 0.5 ng/mL [340]. 

With the use of pipette tip SPE with MonoTip C(18) tips for extraction of MA 

and amphetamine from whole blood (0.1 mL), followed by derivatization with 

trifluoroacetic anhydride, a GC-MS method was developed with LOD of 1.5 

ng/mL for MA and 1.1 ng/mL for amphetamine [341]. An automated LC-MS/MS 

method for the analysis of amphetamines (amphetamine, MA, MDMA, MDA, 

MDEA, PMA, and ephedrine) using 100 and 50 µL of blood and urine, 

respectively, was developed. The LOQs in blood and urine were 2.5 µg/L and 

25 µg/L, respectively, and the runtime per sample was under 11 min [293]. 

A headspace GC-MS method was reported for the determination of GHB in 

serum and urine by converting GHB to GBL in the presence of acid. The LODs 

for serum and urine were 0.16 µg/mL and 0.17 µg/mL, respectively [342]. A 

GC-MS method for quantitative determination of GHB and its precursors [GBL, 

BD] in blood and/or urine samples was reported. GHB and BD were extracted 

with tert-butylmethylether in test tubes preloaded with NaCl followed by acid 

wash, solvent evaporation, and derivatization with MSTFA prior to GC-MS 

analysis. When analyzing GBL, the salting-out step was omitted, and analysis 

was performed with a GC-FID apparatus [343]. 

A UPLC-MS/MS method was validated for the confirmation and quantitation of 

LSD, iso-LSD, N-demethyl-LSD (nor-LSD), and 2-oxo-3-hydroxy-LSD 

(O-H-LSD) [344]. LOQs in blood and urine were 20 pg/mL for LSD and iso-LSD 

and 50 pg/mL for nor-LSD and O-H-LSD. This was the first reported detection 

of O-H-LSD in blood from a suspected LSD user. 

654

3.3.4 Chemical warfare agents 

Organophosphorus nerve agents and sulphur or nitrogen mustard are among 

the most toxic organic compounds used as chemical warfare agents by both 

military and terrorists. An overview of biological markers of exposure to 

chemical warfare agents was reported; metabolites, protein, and/or DNA 

adducts were identified for most nerve agents and vesicants, and validated in 

experimental animals or in a small number of human exposures [345]. 

An analytical method for determining the metabolic hydrolysis products of 

organophosphorus nerve agents, such as sarin (GB) and cyclosarin (GF), has 

been developed using LC-ESI-TOFMS, with an LOD of 2 ng/mL in plasma 

[346], and HILIC-MS/MS down to LOQ of 1 ng/mL in human urine [347]. The 

biomarker ethyl methylphosphonofluoridate (VX-G) of VX was measured in red 

blood cells (RBC) following treatment with fluoride ion using isotope-dilution 

GC-MS/MS, with an absolute detection limit of < 1 pg on column [348]. The 

method was used for analysis of minipig RBC samples following exposure to 

VX vapor [349,350], and concluded that VX-G was a more precise indicator for 

severity of exposure than analysis of acetylcholinesterase (AChE) inhibition 

[349]. 

Four methods were used to monitor the reaction of albumin with soman: 

Progressive inhibition of the aryl acylamidase activity of albumin, the release of 

fluoride ion from soman, 31 P NMR, and MS. The study concluded that 

soman-albumin adducts detected by MS was useful for the diagnosis of soman 

exposure [351]. A fluoride ion-based regeneration assay with isotope dilution, 

followed by large volume injection GC-CI-MS, was developed for the detection 

of soman in rat plasma. With the use of an isotope labeled internal standard, 

the LOD was 5.65 pg (on column) with an average recovery of 93% [352]. 

Nanoparticle-based electrochemical immunosensor provides an opportunity to 

develop field-deployable, sensitive, and quantitative biosensors for monitoring 

exposure to a variety of OP pesticides and nerve agents, by the detection of 

the potential marker phosphorylated acetylcholinesterase (Phospho-AChE). 

Experimental results showed that the developed electrochemical quartz crystal 

microbalance technique can allow the detection of Phospho-AChE in human 

plasma, with a detection limit of 0.020 nM-8.0 pM [353,354]. The AChE 

enzyme activity was investigated using a carbon nanotube-based 

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electrochemical sensor with commercially available purified AChE and ChE in 

saliva obtained from nave rats. It is found that the sensor is very sensitive, with 

an LOD for AChE down to 2 pM [355]. 

An automated on-line pepsin digestion-LC-MS configuration has been 

developed for the rapid determination of chemical warfare agent protein 

adducts, and was demonstrated by the analysis of specific adducts of sarin 

and sulfur mustard to human butyryl cholinesterase and human serum albumin, 

respectively [356]. To identify five hydrolysis degradation products of sulfur 

mustards, namely thiodiglycol, bis(2-hydroxyethylthio)methane, 

1,2-bis(2-hydroxyethylthio)ethane, 1,3-bis(2-hydroxyethylthio)propane, and 

1,4-bis(2-hydroxyethylthio)butane, a RPLC coupled with inductively coupled 

plasma mass spectrometry (ICP-MS) was employed for element-specific sulfur 

detection, and ESI-MS was used to confirm the identification [357]. An SPE 

followed by GCMS was reported for the analysis of nitrogen containing amino 

alcohols, the precursors and degradation products of nitrogen mustards and 

VX in plasma. The LOD was down to 5 ng/mL in the selected ion monitoring 

mode and 0.01 µg/mL in full scan mode [358]. An LC-MS/MS procedure has 

been developed for retrospective diagnosis of exposure to different forms of 

mustard agents [nitrogen mustards (HN-1, HN-2, and HN-3) or sulfur mustard 

(HD)] by detection of the CWA-adducts formed in blood samples, enabling 

LODs of 200 nM of HN-1, 100 nM of HN-2, 200 nM of HN-3, or 50 nM of HD 

[359]. 

Timely diagnosis of exposure to ricin is of primary importance for appropriate 

treatment. A sensitive and specific immunochromatographic test for ricin has 

been developed for identifying ricins prepared from 19 Ricinus cultivars, all 

with a limit of visible detection between 1 and 2.5 ng/mL in buffer [360]. 

Matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) 

with an automated laser firing sequence was used to detect intact ricin from 

solutions containing less than 4 µg/mL of ricin in the presence of other 

endogenous seed proteins [361]. 

Methods involving LC-ICP-MS were examined for the determination of 

phenylarsenic compounds derived from chemical warfare agents, such as 

diphenylarsinic acid (DPAA), phenylarsonic acid, phenylmethylarsinic acid 

(PMAA), phenyldimethylarsine oxide, and diphenylmethylarsine oxide, in urine 

[362]. Analysis of the urine samples of the patients exposed to phenylarsenic 

656

compounds indicated that the main phenylarsenic components were DPAA 

and PMAA. 

3.3.5 Toxic herbs and venoms 

Although many plants and animals contain toxic alkaloids which may be 

dangerous to humans, cases of fatal poisonings are relatively rare. A review of 

toxic alkaloids which are often involved in fatal and non-fatal poisonings, was 

reported [363]. It summarized the symptoms of the intoxications, and the 

methods of detection of the toxic constituents in biological fluids, of aconitine, 

atropine, coniine, colchicine, cytisine, dimethyltryptamine, harmine, harmaline, 

ibogaine, kawain, mescaline, scopolamine, and taxine. Toxicological studies 

constitute an essential part of the effort in developing an herbal medicine into a 

drug product. In the U.S., current regulatory views on issues related to 

requirements and recommendations on various types of nonclinical toxicity 

studies in support of clinical trials, and filing a new drug application for a herbal 

medicine, were discussed [364] . 

Multianalyte procedures were developed for detection and quantification of 

aconitine, atropine, colchicine, coniine, cytisine, nicotine, cotinine, 

physostigmine, and scopolamine in plasma, using LC-APCI-MS and 

LC-ESI-MS/MS, with LOQs in the range of 0.1 – 50 ng/mL [365]. With the 

exception of greater sensitivity and higher identification power, LC-ESI-MS/MS 

had no major advantages over LC-APCI-MS. 

A fatal aconitine poisoning case involving a 25-year-old man in Canada who 

died suddenly due to accidental consumption of Monkshood (Aconitum 

napellus), was reported [366]. LC-MS/MS analysis revealed the presence of 

aconitine, at levels of 3.6 and 149 µg/L in blood and urine, respectively. 

LC-ESI-MS-MS methods were developed for simultaneous determination of 

Aconitum alkaloids in rat plasma following oral administration of Sini decoction 

after basic LLE [367], and in human plasma collected from 18 healthy 

volunteers after intravenous drop infusion of "SHEN-FU" injectable powder, 

using SPE for sample preparation [368]. 

Aristolochic acid I (AA I), a major component of the carcinogenic plant extract 

aristolochic acid (AA), is known to be nephrotoxic, carcinogenic, and 

mutagenic. An HPLC-DAD-fluorescence detection method was developed for 

the analysis of AA I and its metabolites in cell culture medium, for investigating 

657

cytotoxicity of AA I on human liver cells (L-02) [369]. Using LC coupled with a 

hybrid quadrupole time-of-flight mass spectrometry, significant changes of two 

endogenous metabolite markers, kynurenic acid and hippuric acid, were 

detected in the rat urine samples; this metabolic profiling may be useful as an 

indicator for AA exposure before symptoms are observed [370]. 

Heimia salicifolia ("sinicuichi") may cause feelings of exhilaration and an 

alteration of awareness accompanied by bradycardia, muscle relaxation, and a 

pleasant faintness. A case of sinicuichi intoxication was reported [371]. After 

liquid extraction with acetone, five different heimia alkaloids were detected in 

the plant material by LC-MS/MS, and these alkaloids were also detected in the 

patient's serum sample. An LC-MS/MS protocol was optimized for the 

collective detection of gelsemine and related alkaloids from Gelsemium 

elegans [372]. Nine cases of gelsemium poisonings were thus ascertained by 

their diagnostic urine alkaloid profiles, where gelsemine was identified as the 

main urinary marker. A rapid LC-MS/MS/MS method for the analysis of 

alpha-amanitin in serum and liver was described, with calculated MDL of 0.5 

ng/L in serum, and positive identification of alpha-amanitin in serum and liver 

from suspect human and animal intoxications [373]. 

A case of sudden death associated with injection of toad extract which was 

originally believed to be the ring-derivative of amphetamine 'ecstasy' (MDMA) 

was reported [374]. Toxicological analyses of the victim's blood showed low 

levels of bufotenine, while the injected material revealed the presence of 

bufotenine, resibufogenin, cinobufagin, and bufalin. 

3.3.6 Drugs in sports and doping control 

Doping refers to the use of performance-enhancing drugs such as anabolic 

steroids, particularly those that are prohibited under the list published by the 

International Olympics Committee. Some doping substances, however, are 

permitted in low doses (alcohol and caffeine). Another form of doping is blood 

doping, either by blood transfusion or use of the hormone erythropoietin 

(EPO). 

In a study of the use of prescribed medication and nutritional supplements in 

female and male junior, youth, and adult track and field athletes, non-steroidal 

anti-inflammatory drugs (NSAIDs; 0.27 per athlete, n = 884), respiratory drugs 

(0.21 per athlete, n = 682), and alternative analgesics (0.13, n = 423) were 

658

used most frequently [375]. 

A comparative study was made between LLE and direct injection for the 

LC-MS/MS detection of 34 diuretics and 9 other doping agents in urine [376]. 

With a 25-fold dilution of the urine samples, the problems of retention time 

shifting and ion-suppression in direct injection were improved compared with 

the undiluted samples, with results comparable to those obtained from LLE. A 

fully automated method based on UPLC-MS/MS for the analysis of 130 

substances by direct injections of urine samples has been developed for three 

different classes of doping agents: diuretics, central nervous system stimulants 

(CNS stimulants), and opiates [377]. 

Direct analysis of urine samples by UPLC-MS/MS has been developed for 

quantitation of salbutamol [378]. With salbutamol-d6 as internal standard 

followed by dilution with ultrapure water (1:1), the LOQ was 200 ng/mL. A 

quantitative LC-MS/MS method to detect altizide, hydrochlorothiazide and their 

common metabolites (chlorothiazide and 

4-amino-6-chloro-1,3-benzenedisulphonamide) was developed, and the 

concentrations ranges were found to be between 41-239 and 60-287 ng/mL 

after altizide and hydrochlorothiazide administration, respectively [379]. The 

study also showed that 4-amino-6-chloro-1,3-benzenedisulphonamide is an 

important target compound to detect usage of thiazide diuretics. CE coupled 

with field-amplified sample stacking (FASS) has been reported for the 

simultaneous determination of strychnine and brucine residues in human urine, 

with detection limits of 2 and 2.5 ng/mL, respectively [380]. 

The administration of glycerol to endurance athletes results in an increased 

fluid retention and improved performance, particularly under hot and humid 

conditions. A quantitative method was established using a GC/isotope-dilution 

MS-based approach, with an LOD of 0.3 µg/mL [381]. A 200 µg/mL threshold 

for glycerol levels in urine is suggested based on the analysis of 1039 doping 

control samples. An important aspect of doping research is the rapid 

implementation of tests for emerging drugs with potential for misuse into 

routine doping control assays. The mass spectrometric fragmentation of four 

new emerging drug candidates: GW501516 (PPAR-delta-agonists), S-107 and 

JTV-519 (ryanodine-calstabin-complex stabilizers), and S-40503 (selective 

androgen receptor modulators) were studied, and a screening and 

confirmation procedure was established for the analysis of these drugs in 

659

plasma based on LC-MS/MS, with LODs in the range of 0.4-8.3 ng/mL [382]. 

3.3.6.1 Anabolic androgenic steroids 

Anabolic androgenic steroids (AAS) are widely abused by athletes and 

account for the majority of adverse analytical findings reported in doping 

control. Testosterone is the most common AAS abused. Although traditional 

methods allow identification and characterization of testosterone and its 

metabolites in urine, they cannot distinguish between pharmaceutical products 

and endogenous materials. A study summarized the results obtained from the 

doping control analysis for delta(13)C values of selected steroids during the 

period of the 2007 Pan American Games [383]. One urine sample with a 

testosterone/epitestosterone (T/E) ratio of 4.7 was confirmed to be positive 

while the delta values of 5-beta-diol and 5-alpha-diol were 3.8 and 10.8, 

respectively, which exceeded the World Anti-Doping Agency (WADA) limit of 3 

per thousand. A study was reported to determine if consuming a diet low in 

13C plants, such as soy, can be confused with a GC/Combustion (C)/Isotopic 

Ratio Mass Spectrometry (IRMS)-positive test for exogenous testosterone 

[ 384 ]. The study concluded that the carbon isotope ratios of urinary 

testosterone metabolites of vegetarians consuming a diet that is markedly 

depleted of 13C content were lower than that of control subjects, but not low 

enough to result in exceeding the WADA limit. 

Debate on the use of >2 ng/mL of 19-norandrosterone (19-NA) in urine taken 

as prima facie evidence of administration of nandrolone or other 19-norsteroid 

has never stopped. Two reports indicated that it is possible for positive 

detection of 19-NA despite not having taken 19-norsteroid [385,386]. 

Contraceptives containing 19-norandrostenedione as impurity per 1 mg and 5 

mg of norethisterone were administered to female volunteers in doses used for 

menstrual disorders, and resulted in maximal 19-NA concentrations of 9.1 

ng/mL and 63 ng/mL, respectively [385]. The urinary concentrations of 19-NA 

in 1202 female volunteers who were not using anabolic steroids, including 38 

subjects using oral contraceptives containing norethisterone, were studied. 

The results showed that only one sample exceeded the 19-NA reporting 

threshold, having a concentration of 4.1 ng/mL [386]. 28 women participated in 

a study in the excretion of nandrolone metabolites [19-NA, and 

19-noretiocholanolone (19-NE)] in urine, as analysed by GC-MS, revealed that 

urinary concentrations of 19-NA and 19-NE ranged from undetectable levels to 

1.14 and 0.47 ng/mL, respectively [387]. Therefore, a urinary concentration of 

660

2 ng/mL of 19-NA was concluded to be a fair upper acceptable limit in doping 

control tests for female athletes. 

The metabolism of methyltestosterone has been reinvestigated by the use of a 

precursor ion scan method in LC-ESI-MS/MS [388]. Two metabolites have 

been detected using this method, namely epimethyltestosterone and a 

dehydrogenated compound. Comparison with the synthesized reference 

standard revealed that the structure of the dehydrogenated metabolite is 

6-ene-epimethyltestosterone. 6-Ene-epimethyltestosterone was found in all 

samples, demonstrating its applicability in the detection of methyltestosterone 

misuse. 

An approach based on precursor ion scanning of m/z 105, 91, and 77 

corresponded to the methyl tropylium, tropylium, and phenyl ions, respectively, 

for the detection of unknown anabolic steroids, their metabolites and 

prohormones, was proposed [ 389 ]. In general, only compounds with a 

steroidal structure showed a signal at all three selected m/z values, although 

some exceptions have been found. A new doping control screening method 

has been developed using HPLC/orbitrap with in-source collision-induced 

dissociation and atmospheric pressure chemical ionization, for the detection of 

29 compounds, including agents with antiestrogenic activity, beta(2) agonists, 

exogenous anabolic steroids, and other anabolic agents [390]. The mass 

accuracy of this method is better than 2 ppm using an external reference, with 

LODs for all compounds tested better than 100 pg/mL. Ten steroids 

(11-beta-hydroxyandrosterone, 5-alpha-androst-16-en-3-alpha-ol, 

pregnanediol, androsterone, etiocholanolone, testosterone, epitestosterone, 

5-alpha-androstan-3-alpha,17-beta-diol, 

5-beta-androstan-3-alpha,17-beta-diol and dehydroepiandrosterone) were 

measured from a single urine specimen by a GC/thermal conversion 

(TC)/IRMS method for deuterium/hydrogen ratio [391]. For a 20 mL specimen, 

the LODs ranged from 10-15 ng/mL. The applicability of comprehensive 

two-dimensional gas chromatography (GCxGC) for sterol analysis was 

investigated by separation and identification of endogenous sterols in 

standards, and spiked in human urine [ 392 ]. The match quality of 

GCxGC-TOFMS spectra was reported to be superior to that for analysis using 

1D GC-TOFMS for sterols spiked in urine at 10 ng/mL. The LOD for estrone 

was 0.1 ng/mL, while the highest LOD was for 

5-alpha-androstan-3-alpha,11-beta-diol-17-one, epitestosterone, and 

661

cholesteryl butyrate (1 ng/mL). 

Trenbolone and its derivatives such as 17-alpha-methyltrenbolone represent a 

class of highly potent anabolic-androgenic steroids. Based on an enzymatic 

hydrolysis of target compounds, LLE and subsequent LC-MS/MS 

measurement, a method was developed for the analysis of 7 related 

compounds, namely trenbolone, epitrenbolone, methyltrenbolone, 

ethyltrenbolone, propyltrenbolone, 17-ketotrenbolone and altrenogest, with 

LODs in the range of 0.3-3 ng/mL [393]. 

A chemical mixture consisting of antibiotics, antimycotic substances, and 

protease inhibitors was added to urine aliquots fortified with conjugated and 

deuterated steroids and inoculated with nine representative microorganisms to 

compare against urine aliquots without the chemical mixture, by analysis of 

steroids in the free, glucuronide, and sulfate fractions by GC-MS [394]. The 

study concluded that addition of the chemical stabilization mixture to urine 

samples inhibited microorganism growth and prevented steroid degradation at 

37 o C, while some microorganisms induced alterations in the steroid profile of 

the unstabilized samples incubated at 37 o C. 

3.3.6.2 Stimulants 

Stimulants were the first group of compounds prohibited by most international 

sports federations. An Italian study carried out in 2003- 2007 showed that the 

percentage of positive test results range from 1.1 to 2%, with a high incidence 

of stimulants and drugs of abuse; THC metabolites were the most frequently 

found, followed by cocaine metabolites [395]. Doping control concerning 

pesudoephedrine in Poland was studied by comparing results obtained in 

2001-2003 and 2004-2007, revealing that athletes used pseudoephedrine 

more frequently and in high doses in 2004-2007 when this substance was 

permitted by WADA [396]. A method for detection of metamfepramone, 

methylpseudoephedrine, and methcathinone by LC-MS/MS was developed to 

allow analysis of these three analytes by direct injection of 2 µL of urine [397]. 

The method was fully validated with LODs of 2-10 ng/mL for all analytes. 

3.3.6.3 Synthetic protein hormone, blood doping, and gene doping 

Darbepoetin alfa (DPO), an analog of recombinant human erythropoietin 

(rhEPO), is abused as a blood doping agent (along with the latter) in human 

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sports. A method was described for unequivocal identification of DPO in 

human plasma [ 398 ]. The analyte was extracted from plasma by 

immunoaffinity separation with anti-rhEPO antibodies, digested by trypsin 

followed by peptide-N-glycoside F (PNGase F), and analyzed by LC-MS/MS; 

the LOD for DPO was 0.1 ng/mL. Four proposed new markers for detection of 

rhEPO abuse were reported. Preliminary data showed that concentrations of 

asymmetrical dimethylarginine (ADMA), symmetrical dimethylarginine (SDMA), 

citrulline, and arginine in human urine were increased after administration of a 

single intravenous erythropoietin injection, up to 40-270 µg /mL, 40-240 µg/ mL, 

10-60 µg /mL and 12-140 µg/ mL, respectively [399]. 

While samples containing endogenous growth hormone (GH) yield up to four 

isoforms, samples with recombinant GH contain the main 22 kDa spot only. A 

method was reported to separate different isoforms by two-dimensional 

polyacrylamide gel electrophoresis (2-D PAGE) after isolation of GH from 

plasma, using an immunoaffinity purification system which detected discrete 

isoforms of GH from plasma, thereby discriminating endogenous GH from its 

recombinant analog [400]. A mass spectrometric approach was developed to 

determine the nondegraded hormone, gonadotropin-releasing hormone 

(GnRH), in doping control samples with SPE, immunoaffinity purification, and a 

subsequent analysis by LC-ESI-MS/MS [401]. The method offered an LOD of 

about 5 pg/mL, and urinary concentrations of the intact peptide in 

post-administration samples ranged from 20 - 100 pg/mL. 

Doping control analysis of performance-enhancing peptides in urine 

represents a challenging requirement in modern sports drug testing: Low 

doses and short half-lives lead to low concentrations (in fmol/mL range) in 

urine. Immunoaffinity purification with UPLC-MS/MS (high resolution/high 

mass accuracy) of Synacthen from urinary specimens is described and 

validated with LOD of 3 pg/mL [402]. 

A study was conducted on the intra-individual variance of total hemoglobin 

mass (tHb-mass) in elite athletes during a training year as the basis to 

measure tHb-mass as an effective screening parameter for detecting blood 

doping [403]. Results showed that tHb-mass was very low for individuals 

during a training year (

for medical reasons or for blood doping to increase athletic performance are 

potentially exposed to the plasticizer di-(2-ethylhexyl)phthalate (DEHP) found 

in IV bags. A study showed that elevated concentrations of urinary DEHP 

metabolites collected from athletes may suggest illegal blood transfusions, and 

can therefore be used as a qualitative screening measure for blood doping 

[404]. 

With the recent technical improvements in genetic manipulation there is 

mounting concern over the possibility of such technology being abused by 

athletes in their quest for improved performance. A review of the literature was 

conducted to identify the genes and methodologies most likely to be used for 

gene doping, and the technologies that might be used to identify such doping 

[405]. Although there is currently no evidence that gene doping has yet been 

undertaken in competitive athletes, the anti-doping authorities will need to 

remain vigilant in reviewing this rapidly emerging technology. So far, the abuse 

of gene transfer technology in sport is undetectable. However, recent studies 

in somatic gene therapy indicated that long-term presence of transgenic DNA 

(tDNA) following various gene transfer protocols can be found in DNA isolated 

from whole blood using conventional PCR protocols [406]. Application of these 

protocols for the direct detection of gene doping would require almost 

complete knowledge about the sequence of the genetic information that has 

been transferred. A novel single-copy primer-internal intron-spanning PCR 

(spiPCR) procedure that overcomes this difficulty has been developed and 

reported [406]. 

In the post-genome era, genetic- and molecular-based tests are being 

evaluated as potential approaches to detect new and sophisticated forms of 

doping. Transcriptome analysis, in which a tissue's complement of mRNA 

transcripts is characterized, is one such method. The quantity and composition 

of a tissue's transcriptome is highly reflective of milieu and metabolic activity. A 

review has briefly described the current understanding of transcriptomes, 

common methods of global analysis of gene expression, and non-invasive 

sample sources, focusing on the potential to apply this approach in doping 

control [407]. 

3.3.6.4 Horse racing 

Two LC-MS methods with direct injection analysis were developed and 

validated for the quantification and identification of hydrocortisone in equine 

664

urine [408]. To overcome matrix effects, a standard additions calibration 

method was applied. The linear range was 0.25-2.5 µg/mL for both methods. A 

method for the simultaneous detection of bovine, porcine and human insulins, 

as well as the synthetic analogues Humalog (Lilly) and Novolog (Novo Nordisk) 

in equine plasma, capable of detecting at 0.05 ng/mL, was described [409]. 

Administration studies of levamisole in horses were carried out using two 

different levamisole preparations. Aminorex and rexamino, in addition to 

levamisole and 4-phenyl-2-imidazolidinone, were detected in 

post-administration urine and plasma samples [410]. It has been established 

unequivocally that the normal use of levamisole products in horses can lead to 

the presence of aminorex, rexamino, and 4-phenyl-2-imidazolidinone in their 

urine and blood samples. A direct injection method was developed and 

validated for the quantification and identification of salicylic acid by 

LC-QTOFMS after the horse urine samples was 900-fold diluted, and the 

internal standard (4-methylsalicylic acid) was added [411]. The intra- and 

inter-assay precisions of salicylic acid in horse urine were better than 2.5% and 

2.8%, respectively, with accuracy better than 3.3% deviation. A method for 

differentiation and identification of rhEPO and DPO in equine plasma by 

LC-MS/MS was reported [412]. The method involved analyte extraction and 

enrichment by immunoaffinity separation with anti-rhEPO antibodies, dual 

digestion by trypsin and peptide-N-glycosidase F (PNGase F), and analysis by 

LC-MS/MS, with LODs of 0.05 ng/mL for DPO and 0.1 ng/mL for rhEPO. 

3.4 Alternative Specimens 

The use of alternative specimens in the field of toxicology was first described in 

1979, when hair analysis was used to document chronic drug exposure. Since 

then, 'alternative' samples such as oral fluid, hair, and sweat have gained 

tremendous attention, and a large number of papers have been published. 

'Alternative' specimens present a number of advantages over 'traditional' 

samples (e.g., blood, urine, and tissues), namely the facts that their collection 

is non-invasive, their adulteration is difficult, and they may allow suitable 

windows of detection for certain drugs. The main disadvantage is that drugs 

may be present in very low concentrations, and there are limited clinical 

studies to aid in results interpretation. A literature review on the use of 

alternative specimens, with special focus on the main analytical and 

chromatographic problems for the analysis of these specimens, was presented; 

the review included discussion on their advantages and disadvantages over 

traditional samples in documenting drug exposure [413]. 

665

3.4.1 Oral fluid 

Oral fluid is a readily available specimen that can be collected by non-invasive 

procedures and that typically contains many drugs of interest in screening and 

diagnosis. Several reviews have been published regarding to the collection 

techniques and application of drug testing of oral fluid [414], the progress in 

oral fluid collection, screening, confirmation, and interpretation of cannabinoids, 

opioids, amphetamines, cocaine, and benzodiazepines [27], bioanalytical 

procedures in testing abused drugs [415], and as an alternative matrix to those 

of plasma and serum for therapeutic drug monitoring of xenobiotics [416]. In 

general, collection techniques can artificially affect the production of oral fluid 

and its subsequent pH. Nevertheless, it is essential that devices used to collect 

oral fluid are checked to ensure reasonable stability and recovery of absorbed 

drug [27,415,416]. A number of issues are also discussed concerning 

collection device performance, such as the variability in the volume of sample 

collected and its implications in terms of quantitative measurements, and the 

drug recoveries from various devices available in the market [415]. Additional 

considerations that receive attention are the importance of providing complete 

validation data with respect to analyte stability, matrix effects, and the choice of 

collection method [415,416]. 

Recent advances in analytical techniques have enabled the detection of drugs 

and drug metabolites in oral fluid specimens. Roche DAT screening assays for 

amphetamines, cocaine metabolite (i.e., benzoylecgonine), and opiates in oral 

fluid were assessed where good results were obtained for sensitivity and 

specificity (> 94%) compared to LC-MS-MS, with overall agreement of >96% 

[417]. An analytical method using GC-MS to determine THC and its metabolite 

THC-COOH in oral fluid was reported and was used to analyze samples from 

12 suspected abusers [418]. Various methods for the analysis of abused drugs 

in oral fluid were reported using LC-MS/MS method [33, 39,40,41,419 ,420]. In 

general, less than 1 mL of oral fluid is required with LOQs from 0.4-10 ng/mL 

[33, 39,40,41,419,420]. Some methods were able to analyze THC with other 

abused drugs simultaneously [39,420]. The uses of porous silicon assisted 

laser desorption/ionization time-of-flight mass spectrometry (pSi LDI-TOFMS) 

for rapid analysis (on the order of seconds) of MA, MDMA, and THC in 

sub-microliter oral fluid was reported [421]. An LC-MS/MS was developed for 

determination of nicotine, cotinine, trans-3'-hydroxycotinine, and norcotinine in 

0.5 mL of oral fluid, with LOQs of 0.2-1 ng/mL [422]. 

666

The stability of several drugs and metabolites (6-MAM, cocaine, THC, and 

THC-COOH) in oral fluid collected by two oral fluid collection devices, namely 

Cozart Drug Detection System and Intercept, was studied. On average, 

9%-12% 6-MAM was converted to morphine and 26% and 41% cocaine to 

benzoylecgonine. Good recoveries were observed for the acid metabolite of 

THC in both collection devices, whereas THC was always scarcely recovered 

[423]. A study of the stability of methadone in oral fluid for long-term storage 

found that it is sufficiently stable at 4 o C for 2 months, while a decrease was 

observed for its metabolite, EDDP [424]. The pre-analytical stability study of 11 

frequently abused benzodiazepines in an oral fluid collecting device (Dräger 

DCD 5000) found that during the 14 days of storage, major loss of analytes 

was found in the benzodiazepines with nitro group, such as flunitrazepam and 

clonazepam, which could be prevented by storing the sample in methanol after 

collection [425]. 

The greatest current limitation in oral fluid testing is the scarcity of controlled 

drug administration studies to aid in result interpretation. A UPLC-MS/MS 

method was reported for the analysis of EtG in oral fluid using SPE for sample 

clean-up, with an LOQ of 4.4 ng/mL [325]. With this method, the concentration 

of EtG in oral fluid collected 2-14 h after a moderate alcohol intake varied from 

13.3 to 57.7 ng/mL. To assess the correlation between oral fluid and plasma 

samples from patients on antidepressants, an LC-MS/MS method was 

developed for the simultaneous determination of nine antidepressants, 

including venlafaxine and some of their metabolites, in both oral fluid and 

plasma [426]. The ratio of venlafaxine in oral fluid to plasma was calculated 

and showed a high variability for 5 patients on four different occasions 

(coefficient of variance=24.2-69.6%). 

3.4.2 Hair 

Apart from additional information obtained for DFSA and routine drug 

monitoring, analysis of hair for drugs is useful in obtaining a drug history for 

weeks or months prior to death. A 5-year-old male who died after drinking a 

fatal dose of ethanol revealed urine and blood alcohol levels of 0.4 and 0.5 

g/dL, respectively [427]. But, the analysis of the boy’s hair demonstrated EtG 

with concentrations indicating that the boy might have occasionally imbibed 

alcohol prior to death. In another study using GC-NCI-MS/MS, EtG in hair was 

analysed after derivatization with perfluoropentanoic anhydride (PFPA); the 

667

LOD was determined to be 3.0 pg/mg and the LOQ was fixed at 8.4 pg/mg 

using 30-mg hair [329]. Hair analyses of autopsy cases with a history of 

alcoholism and/or lesions caused by alcohol revealed that EtG concentrations 

were in the range of 60-820 pg/mg. With the use of 72 head hair and 15 pubic 

hair samples, a study was reported to collate 6-MAM and morphine 

concentrations in head and pubic hair from heroin users [428]. A statistical 

evaluation demonstrated that the lower, middle, and upper concentration 

ranges of 6-MAM and morphine in head hair were in general higher than those 

found in pubic hair. A qualitative drug screening method involving alkaline 

hydrolysis, purification with mixed-mode SPE followed by LC-TOFMS analysis, 

was developed for analysis of 32 hair samples from deceased drug addicts 

[243]. The results identified 35 different drugs, and confirmed that drugs were 

found in the majority (72%) of the cases; tramadol was predominantly found. A 

study was conducted in the UK for 286 selected cases reported between 2004 

and 2006, where hair was submitted in addition to the routine specimens for 

analysis. The study demonstrated that hair analysis could provide vital 

evidence to understand both the medical cause of death and the 

circumstances surrounding the death [429]. 

The detection of EtG is of interest in both clinical and forensic contexts. EtG is 

incorporated into growing hair, allowing retrospective investigation of chronic 

alcohol abuse. To give more clues on the alcohol use history, several studies 

were conducted in an attempt to differentiate social drinkers from heavy 

drinkers [326,328]. The levels of EtG in heavy drinkers and social drinkers 

using 30-mg samples were determined following cleanup using SPE graphite 

cartridges and LC-MS/MS [326]. With an LOD and LOQ of 4 and 10 pg/mg, 

respectively, hair EtG concentrations in social drinkers were 50 pg/mg for fatal cases with known history of heavy drinking. 98 hair 

samples from volunteers of social drinkers and heavy drinkers, whose ethanol 

daily intake (EDI) was estimated by means of a written questionnaire, were 

analyzed by LC-MS/MS (LOQ: 3 pg/mg) [328]. The EtG level of 27 pg/mg was 

concluded to provide the best compromise between sensitivity (0.92) and 

specificity (0.96) at detecting an EDI of 60 g or higher during the last 3 months. 

Hair specimens from 15 patients after alcohol abuse cessation were 

segmented and analyzed for EtG, with measured concentrations varying from 

8 to 261 pg/mg; good correlation was found with self-reported past alcohol 

consumption [430]. In view of the usage of different cut-off levels of EtG used 

by different laboratories, consensus by the Society of Hair Testing suggested a 

668

cut-off of 30 pg/mg scalp hair (measured in the 0-3 cm proximal segment) to 

differentiate chronic excessive alcohol consumption [431]. Black and white 

patches of fur from LE rats were taken before and after daily intraperitoneal 

injection of ethanol to study the effect of hair pigmentation in fatty acid ethyl 

ester (FAEE) incorporation [432]. The FAEE profiles were similar between 

black and white hair, and so hair pigmentation was concluded not to affect 

FAEE incorporation into hair. To assess the influence of ethanol contamination, 

an in vitro experiment was performed, leaving hair in an atmosphere saturated 

with ethanol vapor for 15 days [433]. Under this stressed experimental 

condition, the spontaneous production of FAEEs was demonstrated by 

analyzing hair daily with an SPME-GC-MS technique, and a constant increase 

of ethyl myristate, palmitate, and stearate was observed. The problem of hair 

contamination that can occur, for example, with ethanol-containing cosmetics 

should be taken into account in interpretation of results. 

GC-MS and LC-MS(MS) are commonly employed in hair drug analysis, but 

other analytical techniques are also used. Using 100 mg of hair sample and 

nalorphine as internal standard, a capillary zone electrophoresis mass 

spectrometry (CZE-MS) method was developed for analysis of common drugs 

of abuse and their metabolites [265]. The LODs were lower than the 0.1 ng/mg 

cut-offs commonly adopted in hair analysis. MALDI-MS was used for the rapid 

detection of cocaine, benzoylecgonine, and cocaethylene in hair. The hair 

samples were required to be incubated in methanol/trifluoroacetic acid for 15 

min at 45 o C prior to MALDI-MS analysis [434]. Surface-activated chemical 

ionization (SACI) was employed for the analysis of cocaine and its metabolite, 

benzoylecgonine, extracted from hair. Following decontamination and acid 

hydrolysis, the sample solutions were diluted (1:10) and directly analyzed by 

LC/SACI-MS [435]. The LODs and LOQs were 0.003 ng/mg and 0.01 ng/mg, 

respectively, for cocaine and 0.02 ng/mg and 0.04 ng/mg for benzoylecgonine. 

Detection of drugs of abuse in hair is also important for monitoring compliance 

with drug abstinence. Therefore, understanding the mechanisms and timeline 

of drug disappearance from hair is critical for clinical and forensic application of 

hair testing. The kinetics of disappearance of cocaine and its metabolite, 

benzoylecgonine, from hair after discontinuation of drug use was evaluated 

using 137 subjects [436]. The results revealed that the median half-life of both 

cocaine and benzoylecgonine was 1.5 months and approximately 3-4 months, 

were required to “pass” hair testing when the analysis was performed in the 

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segment proximal to the scalp. 

An LC-MS procedure has been developed for the determination of 

methylphenidate in hair of treated children [437]. Using 20 mg hair per assay, 

hair methylphenidate concentrations varied from 0.15 to 4.17 ng/mg hair, with 

decreasing drug concentration in distal hair segment. A 2D GC-MS method 

was described for the analysis of fentanyl in hair [438]. With an LOQ of 5 

pg/mg, the fentanyl concentrations detected in four hair samples collected at 

autopsy were 12, 17, 490, and 1930 pg/mg. An assay of citalopram, 

escitalopram, and their demethylated metabolites in 10-mg hair samples has 

been developed using LC-MS/MS, with LOQs of 25 pg/mg for all analytes [439]. 

Neonatal hair samples from two real cases of gestational exposure were 

analyzed using this method to document their in utero exposure to the drugs. 

Hair analyses of two non-fatal cases of intoxication with pesticides, namely 

alachlor and carbofuran, were reported. Alachlor was detected for the 5 

segments in the range of 12 to 136 pg/mg in the first case while carbofuran 

and 3-hydroxycarbofuran were detected at concentrations of 207 and 164 

pg/mg, respectively, for the second case [440]. The chronic exposure of 

different working population groups in Greece to common currently used 

pesticides (diazinon, fenthion, methyl parathion and malathion) and two 

banned pesticides [hexachlorocyclohexane (HCH) and DDT] was studied 

through hair analysis [441]. GC-ECD and GC-MS analyses of the liquid extract 

of the pulverized hair determined that the median concentrations of total HCHs 

and DDTs detected in the three working groups were: 95.0 and 8.9 pg/mg for 

the greenhouse workers, 38.2 and 3.3 pg/mg for the animal breeders, and 24.1 

and 5.2 pg/mg for the open cultivation group. 

Determination of 8 anabolic steroids (methyltestosterone, stanozolol, 

methandienone, nandrolone, trenbolone, boldenone, methenolone, and DHEA, 

which were administered intraperitoneally) in guinea pig hair has been 

developed using LC-MS/MS and GC-MS/MS [442]. The peak concentrations in 

hair were found on days 2-4, except stanozolol, which peaked on day 10. In 

another similar study by analyzing full-length hair samples from volunteers to 

study the physiological concentration of anabolic steroids, the physiological 

concentrations of testosterone were in the range of 0.8-24.2 pg/mg, 0.1-16.8 

pg/mg, and 0.2-11.5 pg/mg in males, females, and children, respectively, and 

were very much lower than the mean values of dehydroepiandrosterone [443]. 

670

The differentiation between systemic exposure and external contamination for 

certain drug groups has been frequently referred to as one of the limitations of 

drug testing of hair. Difficulty in the interpretation arises when metabolites are 

not detected which may either be due to external contamination of the hair, or 

low doses of the drugs used, in particular when the drugs are administered by 

smoking, sniffing or inhalation. Hair samples of eight pre-Columbian mummies 

were analyzed for drugs using GC-MS. Nicotine was detected in 3 of the hair 

samples, with measured concentrations of 11.4-57.5 pg/mg, but all 3 cases 

revealed negative results for cotinine [444]. Thus, an active consumption of 

tobacco could not be confirmed based on the results of the analysis. The 

determination of the ratio of drug(s) in wash residue (W) to that in hair (H), i.e., 

W/H ratio, is proposed to differentiate system exposure to external 

contamination [445]. When the W/H ratio is less than 0.1 or null, it would tend 

to indicate drug use. Where the W/H ratio is above 0.1 but less than 0.5, it is 

likely to indicate possible use possibly combined with a level of external 

contamination. A W/H ratio greater than 0.5 is likely to indicate that the source 

of most of the drug in the wash residue is from external contamination. The 

W/H ratios of 891 results were evaluated over 13 analytes. Between 74 and 

100% of the analytes studied produced W/H ratios less than 0.5, in particular 

for cannabis (93%) and cocaine (95%), where external contamination is more 

likely because of the way the drug is used. Hair samples from cocaine users 

and seized cocaine samples were compared for the possible profiles of 

metabolites/chemical degradation products [446]. It was found that norcocaine, 

cocaethylene and anhydroecgonine methyl ester (each normalised to cocaine) 

were significantly increased in the incorporation group (i.e., hair samples from 

cocaine users). Hair samples were contaminated by rubbing with cocaine 

followed by sweat application, multiple shampoo treatments and storage to 

study criteria to differentiate the use of cocaine from contamination [447]. All 

contaminated samples were correctly identified as contaminated by cutoff, 

benzoylecgonine presence, percent of benzoylecgonine, and/or the wash 

criterion. 

3.4.3 Sweat 

A controlled administration of MDMA to 15 volunteers with histories of MDMA 

use was conducted to understand the excretion of MDMA and metabolites in 

sweat [448]. MDMA was the primary analyte detected, with concentrations up 

to 3007 ng/patch. 35.0% and 9.4% of the samples were positive for MDMA and 

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MDA, respectively, at the 25-ng/patch threshold proposed by SAMHSA. 

Variability in sweat excretion suggests that results should be interpreted 

qualitatively rather than quantitatively. In a similar study for MA, 8 participants 

received four 10-mg oral doses (low dose) of sustained-release S-(+)-MA HCl 

(d-MA HCl) within 1 week, and 5 of them also received four 20-mg (high) doses 

3 weeks later. MA was measurable in sweat within 2 h of dosing. 92.9% (low 

dose) and 62.5% (high dose) of weekly sweat patches were positive, with 

median concentrations of MA being 63.0 and 307 ng/patch, respectively [449]. 

THC excretion in 11 daily cannabis users after cessation of drug use was 

evaluated. Sweat patches worn the first week of continuously monitored 

abstinence had THC above the proposed cutoff concentration of 1 ng 

THC/patch, with mean THC concentrations being 3.85±0.86 ng/patch. Daily 

and weekly sweat patches from seven subjects who were administered oral 

doses of up to 14.8 mg THC/day for five consecutive days were also tested, 

but no daily or weekly patches had THC above the LOQ (i.e., 0.4 ng 

THC/patch) [ 450 ]. Simultaneously quantification of methadone, heroin, 

cocaine and metabolites in sweat was developed by extracting drugs from 

sweat patches with sodium acetate buffer, followed by SPE and quantification 

by GC/MS, with LOQs of 5-10 ng/patch [451]. 

3.4.4 Meconium 

Meconium is a newborn’s first stool. It is formed over the last trimester of 

pregnancy, and thus can represent exposure to drugs over a 16-20 week 

prepartum and provide a crucial indication of neonatal exposure or maternal 

drug use. Simultaneous quantification of 20 drugs in meconium, including 

cocaine, amphetamine, opiate, and nicotine analytes, by LC-MS/MS was 

developed and applied to document drug-exposed neonates [230]. In a study 

of amphetamines and related analytes in meconium, it was found for the first 

time that para-hydroxymethamphetamine (pOHMAMP), norephedrine 

(NOREPH), and 4-hydroxy-3-methoxymethamphetamine were identified in 

meconium from a MA-exposed neonate [452]. However, MA exposure is poorly 

reflected in neonatal meconium; often, maternal self-reported MA use is not 

corroborated by positive results in amphetamines immunoassays of meconium, 

and even if the initial test results are positive, they frequently are not confirmed 

for MA or amphetamine (AMP) by chromatographic analysis. 

Immunoassay-positive and -negative meconium samples were subjected to 

LC-MS/MS to re-analyse for MA metabolites pOHMAMP, 

p-hydroxyamphetamine (pOHAMP), and NOREPH. It was found that 

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pOHMAMP identified 6 additional neonates whose mothers reported MA 

exposure, yet had a meconium sample screened as negative; pOHMAMP was 

more likely to be present if maternal MA use continued into the third trimester 

[453]. Maternal self-reported drug use and/or meconium toxicology results 

were used to evaluate the effects of prenatal MA exposure on infant and child 

development, and also for potential concurrent exposure to cannabis and 

tobacco. A study showed that meconium tests were more likely to be positive if 

the mother reported MA and cannabis use, particularly in the third trimester, 

but less than half of immunoassay-positive amphetamines (31.0%) and 

cannabis (17.9%) meconium results were confirmed by GC-MS [454]. The 

concentrations of nicotine and 4 metabolites (with and without hydrolysis) in 

meconium from tobacco-exposed and nonexposed neonates were studied by 

LC-MS. Nicotine, cotinine, and OH-cotinine were the most prevalent and 

abundant meconium tobacco biomarkers, and a concentration >10 ng/g was 

suggested to discriminate active from passive and nonexposed neonates 

[455]. 

3.4.5 Other specimens 

The analytical data on placenta concentrations of methadone and metabolites 

after controlled drug administration were reported by analyzing the specimens 

using LC-MS obtained from 5 opioid-dependent women receiving methadone 

pharmacotherapy (doses 65-95 mg on the day of delivery) [456]. LC-MS 

methods were reported for the simultaneous determination of buprenorphine 

(BUP) and related metabolites in umbilical cord [457] and placenta [458]. With 

LODs of 0.8 ng/g, authentic umbilical cord and placenta obtained from an 

opioid-dependent pregnant woman receiving BUP pharmacotherapy was 

analyzed. Interestingly, only the metabolites, namely 

norbuprenorphine-glucuronide, buprenorphine-glucuronide and 

norbuprenorphine, were detected. Analysis of an umbilical cord following 

controlled methadone administration and illicit drug use was accomplished 

with SPE followed by LC-MS/MS analysis. With LOQs of 2.5-10 ng/g, the 

sample was found to contain (in ng/g), 40.3 morphine, 3.6 codeine, 442 

benzyolecgonine, 186 methadone and 45.9 EDDP [459]. 

The usefulness of newborn nails for monitoring in utero drug exposure was 

investigated [460]. Cocaine, benzoylecgonine, morphine, methadone, caffeine, 

nicotine, and cotinine were determined in nail samples from the first 3 months 

of life of 25 newborns abandoned immediately after birth (Group 1), and of 33 

673

abies born at the local hospital recruited on a voluntary basis (Group 2). 

Caffeine (n=12), nicotine (n=13), cocaine and benzoylecgonine (n=6), 

morphine (n=4), and methadone (n=5) were found for Group 1 and methadone 

(n=2), caffeine (n=6), and nicotine and/or cotinine (n=6) were detected in 

Group 2. A GC-MS method was developed for the simultaneous detection and 

quantification of four amphetamine-type stimulants and two cannabinoids in 

fingernails [461]. The LOD and LOQ for each compound were lower than 0.056 

and 0.2 ng/mg, respectively, and positive results were obtained from 

specimens of nine suspected MA or cannabis abusers. The concentration 

ranges of MA, AP, and THCCOOH were 0.10-1.41, 0.12-2.64, and 0.20 ng/mg, 

respectively. 

A case showing the presence of morphine in bile (21.3 µg/mL) and hair (4.8 

ng/mg) but not in blood, urine, or the liver of an addict who survived in hospital 

for about 144 h (6 days) was reported [462]. These data would indicate that the 

enterohepatic cycle does not play any role after the last injection, and directly 

confirmed that gall bladder is a storage depot for morphine. 

Forensic toxicological analyses have traditionally focused on the use of blood, 

body fluids, and certain organs in examinations of deaths due to intoxication. 

However, in some situations, putrefaction and contamination make proper 

sampling from tissues impossible, such as in exhumation cases. Postmortem 

viscera and putrefied bone marrow of pesticides (endosulfan and diazion) 

treated rabbits were analyzed by GC- MS. The results showed that mean 

residue levels of diazinon in the putrefied bodies were 3327 mg/kg in putrefied 

viscera and 1783 mg/kg in the bone marrow, while the levels were 0.26 mg/kg 

(endosulfan sulfate), 0.24 mg/kg (alpha and beta isomers of endosulfan), and 

0.1 mg/kg (endosulfan ether) in putrefied samples (putrefied bone marrow and 

putrefied viscera); the cause of death in these cases was acute pesticide 

poisoning [463]. 

3.5 Interpretation of toxicological results 

3.5.1 Drug levels in postmortem specimens 

The interpretation of the analytical results constitutes one of the most difficult 

aspects in forensic toxicology; particularly for postmortem samples where the 

drug concentrations obtained do not necessarily reflect the blood 

concentration at the time of death, due to variations such as the sampling site 

674

and postmortem changes from the time of death to samples taken. In a 

literature review regarding the post-mortem clinical pharmacology, Ferner 

pointed out that there is a lack of adequate studies of the true relationship 

between fatal events and the concentrations that can be measured 

subsequently, noting that without such studies, clinical pharmacologists and 

others should be wary of interpreting post-mortem results [464]. To assist in the 

interpretation of drug levels found, a review of seemingly similar cases can 

provide added information to assist in understanding the role of drugs/poisons 

found under the circumstances presented. 

3.5.1.1 Alcohols and related biomarkers 

A death of a diabetic man where the concentration of ethanol in post-mortem 

blood rose from 0.4 g/L 2 days after autopsy to 3.5 g/L 10 days after autopsy 

was investigated. Further analysis showed no EtG, suggesting postmortem 

formation of ethanol, due possibly to the presence of glucose and insufficient 

amount of fluoride added [465]. The use of biomarkers of ethanol can establish 

whether a deceased had consumed ethanol before death. In a study of 36 

cases where postmortem formation of ethanol was suspected, 19 cases 

showed the presence of EtG and EtS, indicating antemortem ingestion of 

alcohol [466]. Analysis of EtG in hair and phosphatidylethanol (PEth) in femoral 

blood taken from 70 autopsy cases found that 39 cases had EtG levels above 

the cutoff limit (≥30 pg/mg) compared with 29 for PEth (≥0.7 µmol/L), 

suggesting that measurements of EtG in hair may provide improved diagnostic 

information on alcohol abuse [467]. A unique case was reported for analysis 

of ethanol conjugates in the biological material of a 50-year-old known 

alcoholic whose corpse was exhumed 27 years after death [468]. EtG and EtS 

were identified and quantified in the liver, the kidney, and a blood clot, and EtG 

was identified in all hair segments, suggesting antemortem use of alcohol. EtG 

levels in vitreous humor (VH) were compared with blood and urine in 25 

post-mortem cases; VH EtG was in measurable amounts in 19 cases, ranging 

from 0.05 to 1.90 mg/L; in 21 cases in blood (EtG levels: 0.64 to 5.82 mg/L); 

and 17 cases in urine (EtG levels: 0.25 to 623 mg/L); and there was a 

significant relation between VH EtG levels and blood and urine EtG levels 

[321]. 

The measurement of FAEE in human meconium as biomarkers for prenatal 

alcohol exposure was reviewed [469]. 10 articles including 2,221 subjects 

(range per study 4 to 725) identified 455 (20.5%) subjects as prenatal alcohol 

675

exposed using the methods stated in the articles. FAEE levels above the 

studies' respective cutoffs were reported for 502 (22.6%) subjects. The 

prevalence of gestational ethanol exposure and subsequent fetal exposure 

has been assessed in a cohort of mother-infant dyads in a Mediterranean city 

(Barcelona, Spain) by meconium analysis of FAEEs. Of the 353 meconium 

samples analyzed for FAEEs, 159 (45%) samples contained a total amount of 

7 FAEEs ≥ 2 nmol/g, the cutoff for differentiating heavy maternal alcohol 

consumption during pregnancy [470]. In a similar study conducted in Canada, 

meconium FAEE extracted by LLE and SPE prior to analysis with GC-FID and 

GC-MS confirmed that 17 samples were above the 2 nmol/g (7.74-151.28 

nmol/g of FAEE) out of 682 meconium samples [471] 

False-positive EtG screening results by the DRI ® ethyl glucuronide enzyme 

immunoassay caused by chloral hydrate intake was reported, where 

trichloroethyl glucuronide as an important chloral hydrate metabolite was the 

most probable cross reacting substance [ 472 ]. A controlled study was 

conducted using 10 healthy male volunteers in a fasted state to compare the 

sensitivities and detection times for EtG, EtS, and GTOL/5-HIAA, after a single 

ingestion of ethanol [473]. Compared to ethanol testing in urine, the detection 

times for GTOL/5-HIAA were approximately 5 h longer, and for EtG and EtS 

approximately 25 h longer. In another study, the median detection time for an 

elevated urinary GTOL/5-HIAA ratio was 39 h, while EtG was detected for a 

median of 65 h [331]. In a controlled experiment, ingestion of EtG and EtS as 

components of nonalcoholic wine lead to detection of urine EtS only. A positive 

EtG or EtS result after accidental alcohol exposure is unlikely in blood and oral 

fluid [319]. The serum/blood (S/B) concentration ratios for EtG and EtS were 

determined by an LC-MS method for samples taken from patients at admission 

to an alcohol rehabilitation clinic. The median S/B value for EtG was 1.69, and 

that for EtS was 1.30, indicating that the levels of serum EtG and EtS are 

higher than those in the blood [474]. Ten male volunteers consumed ethanol at 

a fixed dose of 0.5 g/kg body weight in a fasted state was used for the 

pharmacokinetic study of ethanol and EtG in blood and urine. EtG reached its 

maximum concentration [C(max)] at a median time of 4 h, a median time of 3 h 

after C(max) for ethanol. The median EtG elimination half-life was 2.2 h, and 

the renal clearance was 8.32 L/h [475]. 

Three corresponding cases of fatal methanol intoxications with different 

survival times were investigated. High methanol levels were determined in the 

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ains, with very low concentrations in femoral venous blood (738 mg/kg vs. 5 

mg/L and 1008 mg/kg vs. 228 mg/L) and other postmortem specimens [476]. 

Toxicological analysis of a fatality involving a woman who ingested an 

unknown quantity of a pine oil-containing product was reported [ 477 ]. 

Postmortem blood, urine, and stomach content levels of 1-alpha-terpineol 

analyzed by GC-MS were 276 mg/L, 0.5 mg/L, and 4.0 g/total contents, 

respectively, and isopropanol levels analysed by GC-FID were 730 mg/dL, 20 

mg/dL, and 1000 mg/dL, respectively, with no acetone detected. 

3.5.1.2 Abused drugs 

A study on the gender differences in pharmacokinetics for opiates using male 

and female Wistar rats showed that the maximal contents of opiates in blood of 

both genders were found in the second measurement time (15 min), and faster 

distribution of opiates from blood to brain in the female compared to male rats 

[ 478 ]. Human subjects, who received 16 mg buprenorphine and 4 mg 

naloxone, were found to have naloxone detected up to 2 h in three subjects 

and up to 4 h in one subject, with mean maximum concentration was 0.139 ± 

0.062 ng/mL. In urine samples taken 24 h after administration, about 55% of 

the daily dose was excreted in urine in either conjugated or unconjugated 

forms of naloxone and nornaloxone [479]. There has been an increased 

awareness of illicit opiate abusers using the narcotic oxymorphone (Opana) by 

inhalation. Two oxymorphone-involved fatalities were analyzed using GC-MS. 

Concentrations of oxymorphone in blood samples of the deceased were 0.05 

mg/L and 0.12 mg/L, respectively [480]. 

A study examining the plasma pharmacokinetics of MDMA and metabolites 

using controlled oral administration of MDMA in young adults found that the 

mean maximum plasma concentrations of 162.9 ± 39.8 and 171.9 ± 79.5 

ng/mL were observed for MDMA and HMMA, respectively, at low dose (1.0 

mg/kg), and that the mean half-lives of MDMA, and HMMA were approximately 

7 to 8 hours, and 11.5 to 13.5 hours, respectively [481]. Three cases of MA 

related fatalities were reported [482,483,484]. In one case, about 3 g of MA 

was swallowed, the MA concentration in the hospital blood taken 

approximately 12 h after ingestion was 3.0 mg/L, and that in the femoral blood 

from autopsy was 30 mg/L [482]. The other two cases involved MA body 

packers that were believed to be poisoned due to MA leakage from the 

packages into the stomach. The MA concentration was found to be 8.6 mg/L in 

plasma 17 h before death, and 63.5 mg/L in the postmortem cardiac blood in 

677

one case [483], while 24.8 mg/L of MA was found in the cardiac blood in the 

other case [484]. Toxicological examination of a patient with a rib fracture from 

a traffic accident who died after being hospitalized for 13 days revealed 0.178 

and 0.051 µg/mL of MA, and amphetamine, respectively, in the heart blood 

[485]. The cause of death was concluded to be circulatory failure due to MA 

poisoning, and hair samples collected during an autopsy suggested that she 

was a MA abuser for at least 3 months before her death. 

An investigation of cocaine-related death cases from 1990 to 2005 in Italy 

revealed that the mean ratios of cocaine/benzoylecgonine in overdose cases 

were 10.28 in the brain and 0.69 in the blood, and were clearly different from 

those found in incidental cases (brain mean was 0.71 and blood mean was 

0.21) [486]. 

Concentrations of THC and its metabolites were measured in plasma samples 

taken from 18 long-term heavy cannabis smokers residing in an in-patient 

research unit for seven days of monitored abstinence [ 487 ]. THC 

concentrations were > 1 ng/mL in nine participants (1.2–5.5 ng/mL) while 

THCCOOH was detected (2.8–45.6 ng/mL) in all participants after 7 days. The 

study suggested that THC presence in plasma for seven days of abstinence 

may not indicate recent use in daily cannabis users. 

The pre-analytical variation of methadone measurements was evaluated and 

compared to the analytical variation [488]. 27 duplicate blood samples from 

routine autopsy cases (mostly drug addicts) were collected from the left and 

right femoral veins and analyzed for the R- and S-enantiomers of methadone 

and EDDP by a chiral LC-MS method. The total measurement uncertainty 

(CVT) was estimated from the pre-analytical variation (derived from 

measurement of right and left femoral blood samples, CVPA), analytical 

variation (analytical coefficient of variance (CV) of the method, CVA), and 

variation related to calibration (CVCal). The pre-analytical variation amounted to 

a CV% of 19-21% for R- and S-methadone, and 30-38% for R- and S-EDDP, 

i.e., considerably larger than the other components. The results concluded that 

the width of the 95%-uncertainty interval (±2 CVT) for a postmortem 

measurement was largely determined by the component of pre-analytical 

variation. 

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3.5.1.3 Other drugs 

Carbon monoxide (CO) intoxication is one of the most common types of 

poisoning in the modern world. A retrospective study based on toxicological 

results of autopsies from 1999 to 2007 in Morocco revealed that 139 out of 

4402 analyses were recorded as acute CO intoxications, and the average 

carbon monoxide concentration was 5.76 ± 4.3 mL/100mL of blood [489]. A 

toxicological study of a deceased found dead in a car equipped with a hose 

leading exhaust from the engine to the interior of the car was reported [490]. 

Analysis revealed a moderately elevated COHb and a high methemoglobin 

saturation level (MetHb) in peripheral blood. The cause of death could possibly 

be attributed to a combination of methemoglobinemia and a moderately high 

COHb concentration. A case of suicidal inhalation of motorbike exhaust, a 

mixture of carbon monoxide (CO) and gasoline vapor, by a 38-year-old female 

was reported [491]. The %COHb determined in blood was 73%. Gasoline 

concentrations in heart blood and vitreous humor were 22.3 and 1.0 mg/L, 

respectively. Simultaneous deaths of two individuals by inhalation of hydrogen 

sulfide were reported [492]. Autopsies of the two patients revealed a partial 

green discoloration of the skin and pulmonary edema with high levels of 

thiosulfate in the blood (0.089 mmol/L and 0.142 mmol/L, respectively). In a 

study to determine toluene exposure, blood toluene levels (BT) and urinary 

hippuric acid (UHA) levels in 65 homeless adolescents were measured. 

Toluene was detected in 91.9% of the investigated population, who presented 

BT levels from 0.5 to 83.7 µg/mL. The study concluded that UHA levels higher 

than 3.0 g/g creatinine indicate intentional exposure to toluene [493]. 

A fatality case involving a 27-year-old woman with epilepsy was studied, with 

femoral blood concentrations of 2.9 mg/kg oxcarbazepine and 66 mg/kg 

10-hydroxycarbazepine, respectively [494]. Reviewing 26 postmortem cases 

involving oxcarbazepine during the period 2001–2006, the 

10-hydroxycarbazepine concentrations ranged from 2.2 to 48 mg/kg; therefore, 

oxcarbazepine intoxication in this case was implicated as a possible cause of 

death. 

Tramadol is an extensively used centrally acting analgesic and 17 forensic 

autopsy cases with fatal unintentional tramadol intoxications among Swedes 

between 1995 and 2005 were examined [495 ]. The median tramadol 

concentration was 2.0 µg/g with a mean age of 44; 14 subjects were identified 

679

with substance abuse histories, with 8 of them being tramadol abusers. A 

young man suspected of suffering from a tramadol overdose was found with 

tramadol concentration of 8 µg/mL on admission in the ICU; the patient 

subsequently died due to severe acute liver failure. Post-mortem toxicology 

revealed tramadol concentrations of 5.2 mg/L in blood, 6.5 mg/g in liver and 

4.5 mg/g in kidney, with desmethyltramadol detected. This is one of a very few 

cases of a lethal poisoning due to tramadol alone [496]. Body fluids and 

tissues in eight doxepin-related deaths were investigated [497]. For two cases 

of fatal doxepin ingestion and another case of high dosage with advanced 

putrefaction, the liver concentration sum (i.e., N-desmethyl metabolite along 

with its parent) may be more useful if a fatal ingestion cannot be clearly 

separated from a person's medication usage. In another study of 9 cases 

related to lethal doxepin intoxication, the concentrations in blood samples from 

peripheral vessels was measured to be 1.5-7.0 mg/L, which is in the lowest 

quarter of the span of lethal concentrations mentioned in the literature - but the 

literature did not specify the location of the blood sample taken [498]. In 

reviewing toxicological reports in Canada, there was an increase in the 

incidence of findings of acetone in decedents who were taking olanzapine over 

the past decade [499]. Olanzapine concentrations were detected in the range 

from

and parathion were reported, with parathion being the most frequently 

detected compound among the four [ 503 ]. The mean concentrations of 

chlorpyrifos, diazinon, malathion and parathion were 0.72, 1.03, 0.82 and 

2.90 mg/L, respectively. A case of a combined suicide by ingestion of a 

pesticide, oxydemeton-methyl (Metasystox ® ) and hanging was reported, 

where the level of dimethyl phosphate (DMP), a stable product of 

oxydemeton-methyl, was determined using a GC-MS procedure to be 

0.10 µg/mL in blood, 1.40 µg/g in liver, and 103.31 µg/mL in gastric contents 

[504]. With no DMP found in urine but a high concentration in gastric contents, 

a close temporal connection between ingestion of poison and hanging is 

assumed. The concentrations of bromadiolone in whole blood and plasma in 

serial samples from a 62-year-old woman, who was repeatedly 

exposed to bromadiolone, using LC-MS/MS were measured. The half-life of 

bromadiolone in blood was estimated to be about 6 days in the initial phase of 

elimination, and about 10-13 days in the terminal phase. The mean 

plasma/blood ratio of bromadiolone was 1.7 ± 0.6. Stability testing of 

bromadiolone in whole blood samples after two cycles of freeze and thaw 

revealed that bromadiolone concentrations decreased [505]. 

Toxicologists should consider that decedents who survived for a relative long 

interval in cases of fatal poisonings may have lower blood concentrations of 

drugs and poisons due to ongoing metabolism while undergoing intensive 

medical treatment. Post-mortem toxicological analysis of a patient of drug 

overdose who received intensive medical treatment for 1 day before death was 

reported. It revealed that concentrations of trazodone, paroxetine and 

7-aminoflunitrazepam in the femoral blood were 5.85 µg/mL, 0.30 µg/mL and 

0.13 µg/mL, respectively, which were all below reported fatal levels. The 

concentrations of paroxetine and trazodone in stomach contents, however, 

were high [506]. The cause of death was concluded to be due to the central 

nervous system depression by the combined effects of trazodone, paroxetine, 

and flunitrazepam. 

Due to continuous consumption of glucose by surviving cells postmortem, 

blood glucose levels decrease rapidly. A study on 3076 cases for vitreous 

glucose and lactate suggested that vitreous glucose alone should be used to 

diagnose hyperglycemia postmortem, and that the limit of 10 mmol/L should 

have a good specificity for diabetic coma, which theoretically would be equal to 

an original blood glucose value of about 26 mmol/L [507]. 

681

Rosano et al. has conducted a study of postmortem blood ethylene glycol and 

glycolic acid in fatal poisoning cases and concluded that measurement of 

glycolic acid is recommended in addition to ethylene glycol for cases of 

ethylene glycol poisoning, as ethylene glycol determination alone may not 

provide a sensitive index for the detection or determination of severity of EG 

poisoning [508]. 

The determination of calcium and sodium content in post-mortem liver, heart, 

and skeletal muscle samples as the biochemical marker defining hypothermia 

as the cause of death was investigated; the results showed that no significant 

differences of calcium content in the tissues of persons who died due to 

hypothermia versus those who died in normothermic conditions [509,510], but 

the concentration of sodium was significantly (p

EtG in blood samples during putrefaction showed that EtG was very unstable 

in blood samples at 30/40 o C without preservatives, but was stable with 

addition of potassium fluoride at room temperature [ 513 ]. In another 

investigation related to stability of EtG and EtS in urine using several 

preservatives, it was found that chlorhexidine, a combination of chlorhexidine, 

ethylparabene and sodium propionate, and boric acid proved to be useful 

preservatives, while EtG degraded in samples doped with thymol, and addition 

of these preservatives did not interfere with the LC-MS/MS analysis [514]. The 

presence of microorganisms capable of ethanol production was checked by 

fermentation tests and the liver was tested for the presence of EtG. It was 

confirmed that there was no EtG formation during bacterial fermentation of 

glucose to ethanol [515]. Even though EtS is claimed to be more stable than 

EtG, a recent study showed possible bacterial degradation of EtS with high 

bacterial density [ 516 ]. Study on post-mortem stability and possible 

post-mortem redistribution of CDT, another alcohol marker, found that CDT 

remains stable for an appreciable time after death and it is not subject to major 

post-mortem redistribution [517]. 

Embalming and formalin fixation are common, and yet they can create 

problems if the only available specimens to be analyzed are formalin-fixed 

tissues. In a study of the stability of cocaine in formalin-fixed liver and brain 

tissues, it was found that in buffered formalin (pH 7.4) cocaine is hydrolyzed to 

benzyolecgonine with half-life time of approximately 7 days, whereas in 

unbuffered formalin (pH approximately 3.5) it is relatively stable over a period 

of 30 days [518]. Eight drugs (sertraline, paroxetine, citalopram, diazepam, 

alprazolam, diphenhydramine, promethazine, and hydrocodone) from four 

different drug classes were evaluated to determine the difference between 

drug concentrations in clamped and blind stick femoral blood, and it was 

concluded that there is no significant difference in the results obtained by these 

two sampling methods [519]. The stability of drugs and metabolites in urine 

after the addition of NaN3 and/or filtration sterilization using an aseptic 

collection kit has been investigated [ 520 ]. In urine that was severely 

contaminated with bacteria, there were significant losses of 

7-aminoflunitrazepam and nitrazepam, and slight decomposition of MA and 

amphetamine at 25 o C. However, such degradation was successfully 

suppressed by the use of the kit, though the use of the kit and NaN3 were 

preferred for 7-aminoflunitrazepam. 

683

As an inherent part of postmortem toxicology, artifacts may arise from the 

antemortem, perimortem and even during specimen collection. In a review 

written by Skopp, he opined that knowledge on the degradation mechanisms, 

breakdown products, and artifactual formation in specimens is of exceptional 

importance to recognize artifacts, and that quantitative analysis is preferably 

done on femoral blood [521]. 

Postmortem redistribution refers to the process of drugs diffusing from tissues 

into blood along a concentration gradient between death and time of specimen 

collection at autopsy. Fentanyl concentrations were measured in postmortem 

specimens collected with mean femoral blood concentrations of 4.6 µg/L and 

17.3 µg/L shortly after death (FB1) and at autopsy (FB2), respectively, and the 

mean fentanyl heart blood (HB) to FB1 ratio of 8.39 - which was higher 

compared with the corresponding HB/FB2 ratio of 3.48 [522]. In another report, 

9 fentanyl-positive cases were identified, and the femoral blood fentanyl 

concentrations ranged from 2.7 to 52.5 µg/L, liver tissue fentanyl ranged from 

37.0 to 179 µg/kg, and heart tissue fentanyl (n = 3) ranged from 52.8 to 179 

µg/kg. Liver tissue to femoral blood ratios ranged from 0.85 to 35.8, and heart 

tissue to femoral blood ratios ranged from 1.9 to 5.4 [523]. Concomitant heart 

and peripheral blood determinations were performed on 40 fatal cases 

involving nordiazepam (20 cases) and bromazepam (20 cases). The mean 

ratios for the heart and peripheral blood concentrations were 0.95 for 

nordiazepam and 0.86 for bromazepam, suggesting that no postmortem 

redistribution was observed for these two benzodiazepines [524]. Postmortem 

redistribution was evident for flecainide, where antemortem plasma obtained 

13 h before death showed a flecainide concentration of 2.5 mg/L, while 

postmortem right and left cardiac blood contained flecainide concentrations of 

13.8 and 44.2 mg/L, respectively [525]. 

3.5.3 Pharmacogenomics 

Pharmacogenomics is one of the first clinical applications of the postgenomic 

era. It promises personalized medicine rather than the established "one size 

fits all" approach to drugs and dosages. Despite initial enthusiasm, the use of 

pharmacogenetics has remained limited. The main reason is the paucity of 

scientific evidence to show that pharmacogenetic testing leads to improved 

clinical outcomes, and the challenge in implementation of pharmacogenomics 

in routine clinical practice [526,527]. Metabolisms of BZP and TFMPP were 

reported to involve the hepatic P450 enzymes CYP2D6, CYP1A2, and 

684

CYP3A4, resulting in inhibited metabolism of other drugs and medicines, as 

well as compromised metabolism in poor metabolisers for CYP2D6. In addition, 

these drugs could cause various drug-drug interactions [528,529]. Furthermore, 

BZP and TFMPP inhibited each other's metabolism [529]. The metabolite ratio 

of amitriptyline (AT), nortriptyline (NT), and its 10-hydroxy metabolites was 

examined by LC-MS in hair samples of 23 white infants after long-term 

administration of AT [530]. High inter-individual variation of the metabolite 

ratios was observed and could be explained for those being CYP2C19 poor 

metabolizers, where the demethylation capabilities were depleted significantly. 

To study the metabolism of naloxone, an LC-MS/MS method was developed 

for determination of naloxone and its metabolite, nornaloxone, in plasma, urine, 

and human liver microsomes. When cDNA-expressed P450s were incubated 

with 20 ng of naloxone, nornaloxone formation was detected for P450s 2C18, 

2C19, and 3A4 [479]. Two cases were studied in which there was a suspicion 

that the patients continued to abuse diazepam, because of repeatedly positive 

urine test results. The measurement of diazepam metabolites in urine showed 

that none of them had in fact ingested diazepam during the study period [531]. 

One of the subjects was found to have a slow metabolism for CYP2C9 as well 

as for CYP2C19. In the second case, there was a possible drug interaction 

between diazepam and zolpidem. 

4. Conclusions 

Phenomenal progress has been achieved in various areas relevant to forensic 

toxicology. The increased availability of more sensitive and specific 

instrumentation has enabled the development of many effective methods for 

the detection of a wider scope of drugs and/or their metabolites at very low 

levels, in both conventional and alternative specimens. 

With the more structured approach towards control over illicit drug use while 

driving, the list of drugs that can be determined is continously expanding. It is 

known that the problem of DUID is far more complex than that of DUI, and that 

it still requires a large amount of research in various areas such as 

epidemiology, behavior, and toxicology. The efforts made in harmonization of 

methods will undoubtedly facilitate more efficient cross-study comparisons in 

future drugged driving research. 

The development of methods for detection of alcohol biomarkers such as EtG, 

EtS and FAEE in oral fluid, urine, blood, vitreous humor, and hair has shed 

685

new light on the investigation of alcohol use: Extension of the detection time for 

alcohol consumption; differentiation of antemortem alcohol use vs. postmortem 

generation due to decomposition; chronic vs. social drinking, etc. It would be 

expected that more laboratories will include alcohol biomarker testing in their 

routine toxicological screening protocols in order to provide additional 

information when required. 

Alternative matrices for analysis of drugs and poisons are becoming popular in 

forensic toxicology. Promising improvements in the reliability of the oral-fluid 

drug screening devices, particularly for the detection of cocaine and more 

recently cannabinoids for DUID, are noted. The scope of drug detection for the 

device, however, is limited and needs to be extended to other common drugs 

of abuse such as ketamine (which is extensively abused in some Asian 

countries). While blood is still regarded as the specimen of choice, the use of 

oral fluid either for roadside testing or even as evidence for prosecution after 

laboratory confirmation of DUID, is increasing. The analysis of hair is 

escalating in various routine drug monitoring programs such as driving license 

re-granting, proof of drug abstinence, and drug-related crimes such as DFSA. 

It should be noted that interpretation of hair test results is difficult due to 

complicated disposition mechanisms; issues of external contamination; and 

limited research available, particularly those in controlled clinical studies. As 

such, more background studies such as controlled administration at clinical 

settings and statistical evaluation of hair drug testing results from drug users, 

are needed. 

The rapid emergence of new drugs of abuse, such as synthetic cannabinoids 

and mephedrone, continues to be one of the major challenges to toxicologists, 

as the unavailability of reference standards (particularly for the corresponding 

metabolites) has hindered the timely development of effective analytical 

methods for their identification. While toxicity data for the new drugs are 

lacking, background studies utilizing data derived from animal models are 

useful in assessing the potential risk to humans, and to determine what 

analytes need to be included in systematic toxicological analysis. This 

information is also valuable to assist governments, medical practitioners, and 

the pharmaceutical industry in formulating policies to reduce the risk to the 

general public. 

The harmonization of quality assurance programs are seen in many toxicology 

laboratories in the adoption of the ISO/IEC 17025 requirements; for instance, 

686

ASCLD/LAB, the world’s largest accrediting body, has become the full member 

signatory to the International Laboratory Accreditation Cooperation (ILAC) 

Mutual Recognition Arrangement (April, 2009). Although the use of alternative 

matrices, such as oral fluid, hair, and/or sweat in toxicological examination is 

becoming more popular, related proficiency tests are surprisingly scarce. The 

proficiency test providers should consider introducing more test programs for 

these specimens in order to ensure the competency of the laboratories offering 

such drug testing services. It is known that the quality of the reference 

materials is of paramount importance to uphold the accuracy of the results 

produced. In view of the limited availability of certified reference materials 

(CRMs), there is an increasing need for the industry to have a long vision 

towards producing CRMs for a wide variety of drugs and/or metabolites in 

related matrices such as blood, oral fluid, and urine. 

The gold standard of using GC-MS in STA for organic drugs and poisons is 

facing a structural change. There has been a gradual shift in many laboratories 

towards the use of LC/MS(MS) due to its improved sensitivity and relatively 

simple sample preparation compared with GC/MS procedures. Moreover, with 

the ability to analyze for a large number of drugs and poisons using LC-MS(MS) 

either using a commercially available database or one developed in-house, 

laboratories are now more willing to switch their screening protocols to 

LC-MS(MS). In addition, the use of isotopic-labeled analytes as internal 

standards is a standard practice for accurate quantitation and to eliminate the 

risk of matrix effects such as ion suppression, particularly in LC-ESI-MS 

analysis. 

With the introduction of more sensitive instruments such as GC-MS/MS and 

LC-MS/MS, increased use of simple sample preparation procedures such as 

dilute and shoot (direct injection) and on-line SPE are reported for relatively 

routine matrices such as urine, oral fluid, and serum. LLE is still the method of 

choice for whole blood, even though some methods have demonstrated the 

applicability of SPME and EME for whole blood extraction. Thus, two of the 

goals in method development will be the replacement of labor intensive 

extraction procedures by automated extraction procedures, and the use of 

smaller sample volumes. 

The collaborative efforts by toxicologists worldwide have filled in many gaps in 

the field through research and development, by sharing of views and 

687

experience in thematic symposia and conferences, and by setting up of new 

professional organization for continual enhancement of the quality and 

knowledge. While there is still a long way to go for the understanding of the 

broad and complicated toxic consequences of myriad chemicals exposure to 

human, toxicologists are ready to take concerted action to tackle the 

challenges ahead. 

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407 Rupert JL. Transcriptional profiling: A potential anti-doping strategy. 

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408 Vonaparti A, Lyris E, Panderi I, Koupparis M, Georgakopoulos C. Direct 

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409 Ho EN, Wan TS, Wong AS, Lam KK, Stewart BD. Doping control analysis 

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410 Ho EN, Leung DK, Leung GN, Wan TS, Wong AS, Wong CH, et al. 

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413 Gallardo E, Queiroz JA. The role of alternative specimens in toxicological 

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414 Drummer OH. Introduction and review of collection techniques and 

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415 Samyn N, Laloup M, De Boeck G. Bioanalytical procedures for 

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416 Mullangi R, Agrawal S, Srinivas NR. Measurement of xenobiotics in 

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417 Crooks CR, Brown S. Roche DAT immunoassay: Sensitivity and 

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418 Choi H, Baeck S, Kim E, Lee S, Jang M, Lee J, et al. Analysis of cannabis 

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419 Simões SS, Ajenjo AC, Franco JM, Vieira DN, Dias MJ. Liquid 

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421 Lowe RD, Guild GE, Harpas P, Kirkbride P, Hoffmann P, Voelcker NH, et 

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422 Shakleya DM, Huestis MA. Optimization and validation of a liquid 

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423 Ventura M, Pichini S, Ventura R, Leal S, Zuccaro P, Pacifici R, et al. 

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424 Fucci N, De Giovanni N. Stability of methadone and its main metabolite in 

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428 Lee S, Cordero R, Paterson S. Distribution of 6-monoacetylmorphine and 

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430 Appenzeller BM, Agirman R, Neuberg P, Yegles M, Wennig R. Segmental 

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433 De Giovanni N, Donadio G, Chiarotti M. Ethanol contamination leads to 

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434 Vogliardi S, Favretto D, Frison G, Ferrara SD, Seraglia R, Traldi P. A fast 

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437 Marchei E, Muñoz JA, García-Algar O, Pellegrini M, Vall O, Zuccaro P, et 

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438 Moore C, Marinetti L, Coulter C, Crompton K. Analysis of pain 

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439 Frison G, Favretto D, Vogliardi S, Terranova C, Ferrara SD. 

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440 Dulaurent S, Gaulier JM, Baudel JL, Fardet L, Maury E, Lachâtre G. Hair 

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441 Tsatsakis AM, Tzatzarakis MN, Tutudaki M. Pesticide levels in head hair 

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442 Shen M, Xiang P, Yan H, Shen B, Wang M. Analysis of anabolic steroids 

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444 Musshoff F, Rosendahl W, Madea B. Determination of nicotine in hair 

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445 Tsanaclis L, Wicks JF. Differentiation between drug use and 

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446 Hoelzle C, Scheufler F, Uhl M, Sachs H, Thieme D. Application of 

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447 Hill V, Cairns T, Schaffer M. Hair analysis for cocaine: Factors in 

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448 Barnes AJ, De Martinis BS, Gorelick DA, Goodwin RS, Kolbrich EA, 

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449 Barnes AJ, Smith ML, Kacinko SL, Schwilke EW, Cone EJ, Moolchan ET, 

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450 Huestis MA, Scheidweiler KB, Saito T, Fortner N, Abraham T, Gustafson 

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451 Brunet BR, Barnes AJ, Scheidweiler KB, Mura P, Huestis MA. 

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452 Kelly T, Gray TR, Huestis MA. Development and validation of a liquid 

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454 Gray TR, LaGasse LL, Smith LM, Derauf C, Grant P, Shah R, et al. 

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455 Gray TR, Magri R, Shakleya DM, Huestis MA. Meconium nicotine and 

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457 Concheiro M, Shakleya DM, Huestis MA. Simultaneous quantification of 

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458 Concheiro-Guisan M, Shakleya DM, Huestis MA. Simultaneous 

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459 de Castro A, Concheiro M, Shakleya DM, Huestis MA. Development and 

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460 Mari F, Politi L, Bertol E. Nails of newborns in monitoring drug exposure 

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461 Kim JY, Cheong JC, Kim MK, Lee JI, In MK. Simultaneous determination 

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462 Tassoni G, Cacaci C, Zampi M, Froldi R. Bile analysis in heroin overdose. 

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463 Akcan R, Hilal A, Daglioglu N, Cekin N, Gulmen MK. Determination of 

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464 Ferner RE. Post-mortem clinical pharmacology. British Journal of Clinical 

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465 Høiseth G, Kristoffersen L, Larssen B, Arnestad M, Hermansen NO, 

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466 Høiseth G, Karinen R, Christophersen A, Mørland J. Practical use of ethyl 

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467 Bebdroth P, Kronstrand R, Helander A, Greby J, Stephanson N, Krantz P. 

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468 Politi L, Morini L, Mari F, Groppi A, Bertol E. Ethyl glucuronide and ethyl 

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469 Burd L, Hofer R. Biomarkers for detection of prenatal alcohol exposure: A 

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472 Arndt T, Gierten B, Güssregen B, Werle A, Grüner J. False-positive ethyl 

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473 Høiseth G, Bernard JP, Stephanson N, Normann PT, Christophersen AS, 

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474 Høiseth G, Morini L, Polettini A, Christophersen AS, Johnsen L, Karinen 

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475 Høiseth G, Bernard JP, Karinen R, Johnsen L, Helander A, 

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476 Andresen H, Schmoldt H, Matschke J, Flachskampf FA, Turk EE. Fatal 

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477 Martz W. A lethal ingestion of a household cleaner containing pine oil and 

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478 Djurendic-Brenesel M, Mimica-Dukic N, Pilija V, Tasic M. Gender-related 

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479 Fang WB, Chang Y, McCance-Katz EF. Moody DE. Determination of 

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480 McIntyre IM, Sherrard JL, Nelson CL. Case report: 

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481 Kolbrich EA, Goodwin RS, Gorelick DA, Hayes RJ, Stein EA, Huestis MA. 

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482 Kiely E, Lee CJ, Marinetti L. A fatality from an oral ingestion of 

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483 Takekawa K, Ohmori T, Kido A, Oya M. Methamphetamine body packer: 

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484 Li RB, Guan DW, Zhu BL, Zhang GH, Zhao R. Death from accidental 

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485 Ago M, Ago K, Ogata M. Determination of methamphetamine in sudden 

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486 Bertol E, Trignano C, Di Milia MG, Di Padua M, Mari F. Cocaine-related 

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487 Karschner EL, Schwilke EW, Lowe RH, Darwin WD, Harning RI, Cadet 

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488 Linnet K, Johansen SS, Buchard A, Munkholm J, Morling N. Dominance 

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489 Ait El Cadi M, Khabbal Y, Idrissi L. Carbon monoxide poisoning in 

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490 Vevelstad M, Morild I. Lethal methemoglobinemia and automobile 

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491 Martínez MA, Ballesteros S. Suicidal inhalation of motorbike exhaust: 

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492 Ago M, Ago K, Ogata M. Two fatalities by hydrogen sulfide poisoning: 

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493 Thiesen FV, Noto AR, Barros HM. Laboratory diagnosis of toluene-based 

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494 Linnet K, Steentoft A, Simonsen KW, Sabers A, Hansen SH. An 

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495 Tjäderborn M, Jönsson AK, Hägg S, Ahlner J. Fatal unintentional 

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497 Gronewold A, Dettling A, Haffner HT, Skopp G. Doxepin and nordoxepin 

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498 Dettling A, Haffner HT, Wehner HD. The evaluation of doxepin 

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499 House CJ. Olanzapine-induced hyperglycemic ketoacidosis and 

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500 Centini F, Fiore C, Riezzo I, Rossi G, Fineschi V. Suicide due to oral 

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501 Kirschbaum KM, Musshoff F, Madea B. Unclear loss of consciousness 

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502 Kłys M, Rojek S, Rzepecka-Woźniak E. Neonatal death following 

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503 Park MJ, In SW, Lee SK, Choi WK, Park YS, Chung HS. Postmortem 

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504 Gottzein AK, Musshoff F, Doberentz E, Madea B. Combined suicide by 

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505 Vindenes V, Karinen R, Hasvold I, Bernard JP, Mörland JG, 

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506 Kinoshita H, Nishiguchi M, Kasuda S, Takahashi M, Ouchi H, Minami T, et 

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507 Zilg B, Alkass K, Berg S, Druid H. Postmortem identification of 

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508 Rosano TG, Swift TA, Kranick CJ, Sikirica M. Ethylene glycol and glycolic 

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509 Jakubeniene M, Irnius A, Chaker GA, Paliulis JM, Bechelis A. 

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510 Jakubeniene M, Chaker GA, Becelis A, Malakiene D, Raudys R. 

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511 Saussereau E, Lacroix C, Cattaneo A, Mahieu L, Goulle JP. Hair and 

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512 Peters FT. Stability of analytes in biosamples - An important issue in 

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513 Høiseth G, Karinen R, Johnsen L, Normann PT, Christophersen AS, 

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514 Thierauf A, Serr A, Halter CC, Al-Ahmad A, Rana S, Weinmann W. 

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515 Appenzeller BMR, Schuman M, Wennig R. Was a child poisoned by 

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516 Halter CC, Laengin A, Al-Ahmad A, Wurst FM, Weinmann W, Kuemmerer 

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517 Rainio J, De Paoli G, Druid H, Kauppila R, De Giorgio F, Bortolotti F, et al. 

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518 Viel G, Nalesso A, Cecchetto G, Montisci M, Ferrara SD. Stability of 

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519 Hargrove VM, McCutcheon JR. Comparison of drug concentrations taken 

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520 Zaitsu K, Miki A, Katagi M, Tsuchihashi H. Long-term stability of various 

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521 Skopp G. Artifacts are an inherent part of postmortem toxicology. TIAFT 

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522 Olson KN, Luckenbill K, Thompson J, Middleton O, Geiselhart R, Mills 

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523 Luckenbill K, Thompson J, Middleton O, Kloss J, Apple F. Fentanyl 

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524 Pos Pok PR, Haddouche D, Mauras M, Kuhlmann E, Burle J, Salmon T, 

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525 Yoshitome K, Miyaishi S, Yamamoto Y, Ishizu H. Postmortem increase of 

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526 Swen JJ, Wilting I, de Goede AL, Grandia L, Mulder H, Touw DJ, et al. 

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527 Swen JJ, Huizinga TW, Gelderblom H, de Vries EG, Assendelft WJ, 

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528 Antia U. 'Party pill' drugs--BZP and TFMPP. The New Zealand Medical 

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529 Antia U, Tingle MD, Russell BR. Metabolic interactions with 

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530 Thieme D, Rolf B, Sachs H, Schmid D. Correlation of inter-individual 

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and CYP2D6 polymorphisms. International Journal of Legal Medicine 

2008 Mar; 122 (2): 149-155. 

531 Lennestål R, Lakso HA, Nilsson M, Mjörndal T. Urine monitoring of 

diazepam abuse- New intake or not? Journal of Analytical Toxicology 

2008 Jul-Aug; 32(6): 402-407. 

734

Drug Evidence 


U.S. Department of Justice 

Drug Enforcement Administration 

Office of Forensic Sciences 

700 Army Navy Drive, W-7342 

Arlington, Virginia 22202 USA 

(Coordinating Office) 

and 

Special Testing and Research Laboratory 

22624 Dulles Summit Court 

Dulles, Virginia 20166 USA 

(Coordinating Laboratory) 

735


I. ROUTINE AND IMPROVED ANALYSIS OF ABUSED SUBSTANCES  740  

1.2  Scientific Working Group for Forensic Analysis of Illicit Drugs:  742  

1.1.  Amphetamine (including substituted amphetamines), Methamphetamine, and  

  Dimethylamphetamine (see Phenethylamines, and Methylenedioxyamphetamines): 743  

1.3  Barbiturates (including Quaaludes):  746  

1.4  Benzodiazepines:  747  

1.5  Dimethoxyphenethylamines:  748  

1.6  Chlordiazepoxide:  749  

1.7  Clenbuterol:  749  

1.8  Cocaine:  750  

1.9  Emerging Substances (including MDPV and “FLY” compounds)  753  

1.10  Ergot Alkaloids (see also LSD):  755  

1.11 Fentanyl(s):  756  

1.12  Flos Daturae:  757  

1.13  Fluoxetine (Prozac):  757  

1.14  Heroin:  757  

1.15  gammaHydroxybutyric Acid (GHB), gammabutyrolactone (GBL) and 1,4butanediol  

  (BD):  759  

1.16  Human Growth Hormone (HGH):  760  

1.17  Ketamine:  760  

1.18  Khat:  760  

1.19  Kratom (Also mitragynine):  761  

1.20  LSD:  762  

1.21  Marijuana and Related Cannabinoids:  762  

1.22  Mescaline/Peyote:  765  

1.23  Methadone:  766  

736

1.24  Methcathinone (Also methcathinonerelated compounds):  766  

1.25  Methylenedioxyamphetamines and Related Compounds:  767  

1.26  Morphine, Codeine, and Related Opium Alkaloids:  769  

1.27  Opiate Alkaloids (Also Opioids):  770  

1.28  Opium (and Opium Poppies):  770  

1.29  Phenethylamines (including mixtures of Amphetamines, Methylenedioxy 

  amphetamines, and Related Compounds):  774  

1.30  Piperazines:  774  

1.31  Polydrug:  776  

1.32  Psilocybin Mushrooms, Psilocybin, and Psilocin:  776  

1.33  Salvia Divinorum:  777  

1.34  Scopolamine:  778  

1.35  Steroids:  778  

1.36  Synthetic Cannabimimetics (Also “Spice”):  780  

1.37  (Designer) Tryptamines (see also Psilocybin):  781  

1.38  Zaleplon:  784  

1.39  Zolpidem:  784  

1.40  Zopiclone:  784  

1.41  Miscellaneous:  785  

2.  SYNTHESIS AND/OR CULTIVATION OF ABUSED SUBSTANCES, THEIR PRECURSORS,  

  AND ESSENTIAL CHEMICALS  785  

2.1  Production of Abused Substances and/or their Precursors and Essential Chemicals:  786  

3.  CLANDESTINE LABORATORIES ‐ APPRAISALS AND SAFETY  790  

3.1  General  790  

3.2  Clandestine Laboratory Appraisals and Safety:  791  

3.3  Safety Issues  Case Reports:  791  


737

4  REFERENCE DRUG STANDARDS AND TOTAL SYNTHESES  792  

4.1  General  792  

5.  SOURCE DETERMINATION OF DRUGS (IMPURITY PROFILING) AND COMPARATIVE  

  ANALYSES  793  

5.1  Amphetamine(s):  794  

5.2  Cocaine:  794  

5.3  Heroin:  795  

5.4  Marijuana:  796  

5.5  Methamphetamine:  797  

5.6  3,4Methylenedioxymethamphetamine:  799  

5.7  MultiDrug and Miscellaneous:  800  

6.  ANALYSIS OF NON‐CONTROLLED PHARMACEUTICALS, PSEUDO‐DRUGS,  

  ADULTERANTS, DILUENTS, AND PRECURSORS  801  

6.1  Ephedra, Ephedrine, and/or Pseudoephedrine and Related Compounds:  801  

6.2  Phenylpropanolamine:  803  

6.3  Other Adulterants/Diluents (including mixtures containing Ephedrine and/or  

  Pseudoephedrine):  804  

6.4  Theophylline:  804  


7.  NEW AND/OR IMPROVED INSTRUMENTAL TECHNIQUES  805  

7.1  Capillary Electrophoresis (and Related Techniques, including Tandem Techniques):  805  

7.2  Extraction Techniques:  807  

7.3  Gas Chromatography (and Tandem GC Techniques):  808  

7.4  HighPerformance Liquid Chromatography (and tandem HPLC techniques):  810  

7.5  Inductively Coupled Plasma Mass Spectrometry (ICPMS, Also ICPOES):  812  

7.6  Infrared and Near Infrared Spectroscopy (including terahertz spectroscopy):  813  

7.7  Ion Mobility Spectrometry:  816  

738

7.8  Mass Spectrometry (including ambient pressure techniques and Isotope Ratio):  816  

7.9  Microchip Technology:  821  

7.10  Nuclear Magnetic Resonance Spectroscopy:  821  

7.11  Raman Spectroscopy:  823  

7.12  Solid Phase MicroExtraction (Headspace Techniques and Solvent Analysis):  824  

7.13  Thin Layer Chromatography:  825  

7.14  XRay based Techniques:  826  


8.  PORTABLE DETECTION AND ANALYTICAL INSTRUMENTATION  827  

8.1 General  828  

9. MISCELLANEOUS  829  

9.1  Beverage Analysis:  829  

9.2  Chemometrics:  829  

9.3  Counterfeit Drugs:  830  

9.4  Dragon’s Blood:  831  

9.5  Drugs on Currency:  831  

9.6  Education:  832  

9.7  Legal Issues:  832  

9.8  Quality Assurance:  833  

9.9  Sampling Plans:  833  

9.10  Toolmarks:  834  

9.11  Other:  834  

739

I. Routine and Improved Analysis of Abused Substances 

Issue: 

Improved methods of analysis, i.e., faster, more discriminatory, more sensitive, 

less costly, etc., are needed for all abused substances. Additionally, standard 

analytical data are required for previously unknown or rarely encountered 

substances and/or new homolog or analog (i.e., "designer"-type) drugs. 

Solution: 

Drug seizures and clandestine laboratory operations are continuously monitored 

to provide a comprehensive overview of new developments. Ongoing research 

in the forensic community, as well as in the general field of analytical chemistry, 

provide new and/or improved methods of analysis for both routine and 

specialized analyses of seized drugs. Reports providing standard analytical data 

for new drugs of abuse and/or improved analytical protocols for known drugs of 

abuse are generated for the forensic and enforcement communities. 

1. Bakouri E, Tsitsimpikou C, Tzifa R, Potamianos S, Tarli A, Vakka 

C, Pantilieri E. The general chemical state laboratory of Greece and its 

role in forensic drug analysis. TOXICOLOGY LETTERS 2008;180:S156- 

S157. 

2. Bell S. Forensic chemistry. ANNUAL REVIEW OF ANALYTICAL 

CHEMISTRY 2009;2:297-319. 

3. Berbatis CG, Sunderland VB, Dhaliwal SS. Linked electronic medication 

systems in community pharmacies for preventing pseudoephedrine 

diversion: A review of international practice and analysis of results in 

Australia. DRUG AND ALCOHOL REVIEW 2009;28(6):586-591. 

4. Bjornsdottir I, Almarsdottir AB, Traulsen JM. The lay public's explicit and 

implicit definitions of drugs. RESEARCH IN SOCIAL & ADMINISTRATIVE 

PHARMACY 2009;5(1):40-50. 

5. Bornmann L, Nast I, Daniel HD. Do editors and referees look for signs of 

scientific misconduct when reviewing manuscripts? A quantitative content 

analysis of studies that examined review criteria and reasons for accepting 

and rejecting manuscripts for publication. SCIENTOMETRICS 

2008;77(3):415-432. 

6. Brettell TA, Butler JM, Almirall JR. Forensic science. ANALYTICAL 

CHEMISTRY 2009;81(12):4695-4711. 

7. Caulkins JP. Price and purity analysis for illicit drug: Data and conceptual 

issues. DRUG AND ALCOHOL DEPENDENCE 2007;90:S61-S68. 

8. Clark RL, Johnston BF, Mackay SP, Breslin CJ, Robertson MN, Sutcliffe 

OB, Dufton MJ, Harvey AL. The drug discovery portal: A computational 

platform for identifying drug leads from academia. CURRENT 

PHARMACEUTICAL DESIGN 2010;16(15):1697-1702. 

9. Cody JT. Hallucinogens. HANDBOOK OF ANALYTICAL SEPARATIONS 

2008;6:175-201. 

740

10. Nic Daeid N, Buchanan HAS, Savage KA, Fraser JG, Cresswell SL. 

Recent advances in the application of stable isotope ratio analysis in 

forensic chemistry. AUSTRALIAN JOURNAL OF CHEMISTRY 

2010;63(1):3-7. 

11. Fazey C. International policy on illicit drug trafficking: The formal and 

informal mechanisms. JOURNAL OF DRUG ISSUES 2007;37(4):755-779. 

12. Fersht A. The most influential journals: Impact factor and Eigenfactor. 

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE 

UNITED STATES OF AMERICA 2009;106(17):6883-6884. 

13. Gorun G, Ceausu M, Francisc A, Curca GC. Thanathogenesis due to inert 

chemical excipients in illicit drugs: Case report and literature review. 

ROMANIAN JOURNAL OF LEGAL MEDICINE 2008;16(3):181-186. 

14. Holden C. Forensic science needs a major overhaul, panel says. 

SCIENCE 2009;323(5918):1155. 

15. Houck MM. Foresight: A business approach to improving the nation’s 

forensic laboratories. 2008 American Academy of Forensic Sciences 

Annual Meeting. 

16. Hu C, Feng Y, Xue J, Chong X, Cheng S. Consideration and science 

development of the fast drug analysis system. YAOWU FENXI ZAZHI 

2008;28(4):647-655. 

17. Kasprzyk-Hordern B, Dinsdale RM, Guwy AJ. Illicit drugs and 

pharmaceuticals in the environment - Forensic applications of 

environmental data. Part 1: Estimation of the usage of drugs in local 

communities. ENVIRONMENTAL POLLUTION 2009;157(6):1773-1777. 

18. Kasprzyk-Hordern B, Dinsdale RM, Guwy AJ. Illicit drugs and 

pharmaceuticals in the environment - Forensic applications of 

environmental data, Part 2: Pharmaceuticals as chemical markers of 

faecal water contamination. ENVIRONMENTAL POLLUTION 

2009;157(6):1778-1786. 

19. Kasprzyk-Hordern B, Dinsdale RM, Guwy AJ. The removal of 

pharmaceuticals, personal care products, endocrine disruptors and illicit 

drugs during wastewater treatment and its impact on the quality of 

receiving waters (vol 43, pg 363, 2009). WATER RESEARCH 

2010;44(6):2076-2076. 

20. Kolosov CA. Evaluating the public interest: Regulation of industrial hemp 

under the controlled substances act. UCLA LAW REVIEW 

2009;57(1):237-274. 

21. Leicach S, Margarita YG, Hugo C. Amines. RESEARCH METHODS IN 

PLANT SCIENCES: ALLELOPATHY 2009;7:291-314. 

22. Liang BA, Mackey T. Searching for safety: Addressing search engine, 

website, and provider accountability for illicit online drug sales. 

AMERICAN JOURNAL OF LAW & MEDICINE 2009;35(1):125-184. 

23. Marone P. Strengthening forensic science in the united states: A path 

forward. Mid-Atlantic Association of Forensic Scientists Annual Meeting: 

2009. 

741

24. Maurer HH. Forensic analysis on the cutting edge - new methods for trace 

evidence analysis. JOURNAL OF THE AMERICAN SOCIETY FOR MASS 

SPECTROMETRY 2008;19(3):R1. 

25. McCambridge J, Winstock A, Hunt N, Mitcheson L. Five year trends in use 

of hallucinogens and other adjunct drugs among UK dance drug users. 

EUROPEAN ADDICTION RESEARCH 2007;13(1):57-64. 

26. McKetin R. The holy grail of supply control: Have we found the evidence? 

ADDICTION 2009;104(3):455-456. 

27. Michels II, Fang YX, Zhao D, Zhao LY, Lu L. Comparison of drug abuse in 

Germany and China. ACTA PHARMACOLOGICA SINICA 2007;28:1505- 

1518. 

28. Mionel O. The drug geopolitics in South America. METALURGIA 

INTERNATIONAL 2008;13(6):71-75. 

29. Paoli L, Greenfield VA, Charles M, Reuter P. The global diversion of 

pharmaceutical drugs. ADDICTION 2009;104(3):347-354. 

30. Paoli L, Rabkov I, Greenfield VA, Reuter P. Tajikistan: The rise of a narcostate. 

JOURNAL OF DRUG ISSUES 2007;37(4):951. 

31. Poret S. An optimal anti-drug law enforcement policy. INTERNATIONAL 

REVIEW OF LAW AND ECONOMICS 2009;29(3):221-228. 

32. Reid G, Devaney ML, Baldwin S. Drug production, trafficking and trade in 

Asia and Pacific Island countries. DRUG AND ALCOHOL REVIEW 

2006;25(6):647-650. 

33. Robertson J. Research front essay: Forensic chemistry. AUSTRALIAN 

JOURNAL OF CHEMISTRY 2010;63(1):1-2. 

34. Sanderson K. Forensic labs warn of deuterated drug threat. NATURE 

2009;458(7240):817 

35. Storti CC, De Grauwe P. Globalization and the price decline of illicit drugs. 

INTERNATIONAL JOURNAL OF DRUG POLICY 2009;20(1):48-61. 

36. Thevis M, Schaenzer W. Emerging drugs - Potential for misuse in sport 

and doping control detection strategies. MINI-REVIEWS IN MEDICINAL 

CHEMISTRY 2007;7(5):531. 

37. Yasuda I, Takahashi M. Analysis of illegal drugs. BUNSEKI 2008(1):26-31. 

38. Yasuda I. The identification of illegal drugs and the change of circulation 

products. TOKYO-TO KENKO ANZEN KENKYU SENTA KENKYU 

NENPO 2007;58:37-45. 

39. Zwingenberger S, Pietsch J, Hommola A, Dressler J. Illegal drug-related 

deaths in East Germany between 1995 and 2004. FORENSIC SCIENCE 

INTERNATIONAL 2010;199(1-3):58-62. 

1.2 Scientific Working Group for Forensic Analysis of Illicit Drugs: 

40. Jackson LC. SWGDRUG update 2009. Mid-Atlantic Association of 

Forensic Scientists Annual Meeting: 2009. 

41. Santos NA. The Scientific Working Group for the Analysis of Seized Drugs 

(SWGDRUG). 2008 American Academy of Forensic Sciences Annual 

Meeting. 

742

1.1. Amphetamine (including substituted amphetamines), 

Methamphetamine, and Dimethylamphetamine (see 

Phenethylamines, and Methylenedioxyamphetamines): 

42. Andreasen MF, Lindholst C, Kaa E. Adulterants and diluents in heroin, 

amphetamine, and cocaine found on the illicit drug market in Aarhus, 

Denmark. OPEN FORENSIC SCIENCE JOURNAL 2009;2:16-20. 

43. Awad T, Belal T, DeRuiter J, Kramer K, Clark CR. Comparison of GC-MS 

and GC-IRD methods for the differentiation of methamphetamine and 

regioisomeric substances. FORENSIC SCIENCE INTERNATIONAL 

2009;185(1-3):67-77. 

44. Awad T, Clark CR, DeRuiter J. GC-MS analysis of acylated derivatives of 

the side-chain regioisomers of 4-methoxy-3-methyl-phenethylamines 

related to methylenedioxymethamphetamine. JOURNAL OF 

CHROMATOGRAPHIC SCIENCE 2007;45(8):477-485. 

45. Belal T, Awad T, Clark CR, DeRuiter J. GC-MS evaluation of a series of 

acylated derivatives of 3,4-methylenedioxymethamphetamine. JOURNAL 

OF CHROMATOGRAPHIC SCIENCE 2009;47(5):359-364. 

46. Bell SEJ, Fido LA, Sirimuthu NMS, Speers SJ, Peters KL, Cosbey SH. 

Screening tablets for DOB using surface-enhanced Raman spectroscopy. 

JOURNAL OF FORENSIC SCIENCES 2007;52(5):1063-1067. 

47. Bijlsma L, Sancho JV, Pitarch E, Ibanez M, Hernandez F. Simultaneous 

ultra-high-pressure liquid chromatography-tandem mass spectrometry 

determination of amphetamine and amphetamine-like stimulants, cocaine 

and its metabolites, and a cannabis metabolite in surface water and urban 

wastewater. JOURNAL OF CHROMATOGRAPHY A 2009;1216(15):3078- 

3089. 

48. Blachut D, Wojtasiewicz K, Czarnocki Z, Szukalski B. The analytical profile 

of some 4-methylthioamphetamine (4-MTA) homologues. FORENSIC 

SCIENCE INTERNATIONAL 2009;192(1-3):98-114. 

49. Boles TH, Wells MJM. Analysis of amphetamine and methamphetamine 

as emerging pollutants in wastewater and wastewater-impacted streams. 

JOURNAL OF CHROMATOGRAPHY A 2010;1217(16):2561-2568. 

50. Bozenko JS. Clandestine enantiomeric enrichment of d-methamphetamine 

via tartaric acid resolution. JOURNAL OF THE CLANDESTINE 

LABORATORY INVESTIGATING CHEMISTS ASSOCIATION 

2008;18(3):4. 

51. Callaghan RC, Cunningham JK, Victor JC, Liu LM. Impact of Canadian 

federal methamphetamine precursor and essential chemical regulations 

on methamphetamine-related acute-care hospital admissions. DRUG AND 

ALCOHOL DEPENDENCE 2009;105(3):185-193. 

52. Chiadmi F, Schlatter J. Crystal meth: A particular form of 

methamphetamine. PRESSE MEDICALE 2009;38(1):63-67. 

743

53. Chintalova-Dallas R, Case P, Kitsenko N, Lazzarini Z. Boltushka: A 

homemade amphetamine-type stimulant and HIV risk in Odessa, Ukraine. 


54. Chung LW, Lin KL, Yang TCC, Lee MR. Orthogonal array optimization of 

microwave-assisted derivatization for determination of trace amphetamine 

and methamphetamine using negative chemical ionization gas 

chromatography-mass spectrometry. JOURNAL OF 

CHROMATOGRAPHY A 2009;1216(18):4083-4089. 

55. Cody JT. Amphetamines. HANDBOOK OF ANALYTICAL SEPARATIONS 

2008;6:127-174. 

56. Coxon A, Mills S. Tackling methamphetamine in New Zealand. JOURNAL 

OF THE CLANDESTINE LABORATORY INVESTIGATING CHEMISTS 

ASSOCIATION 2008;20(1):9. 

57. Cunningham JK, Liu LM, Callaghan R. Impact of US and Canadian 

precursor regulation on methamphetamine purity in the United States. 

ADDICTION 2009;104(3):441-453. 

58. Cunningham JK, Liu LM. Impact of methamphetamine precursor chemical 

legislation, a suppression policy, on the demand for drug treatment. 

SOCIAL SCIENCE & MEDICINE 2008;66(7):1463-1473. 

59. Diozan D, Baheri T, Pournaghi-Azar MH. Development of electro solidphase 

microextraction and application to methamphetamine analysis. 

CHROMATOGRAPHIA 2007;65(1-2):45. 

60. Dobkin C, Nicosia N. The war on drugs: Methamphetamine, public health, 

and crime. AMERICAN ECONOMIC REVIEW 2009;99(1):324-349. 

61. Fu Q, Liao L-c, Chen L-l, Yan Y-y, Yang L, Hou J-h, Chen Y. Detecting 

methamphetamine and amphetamine with high performance liquid 

chromatography. SICHUAN DAXUE XUEBAO, YIXUEBAN 

2007;38(6):1025-1028. 

62. Goh CY, van Bronswijk W, Priddis C. Rapid nondestructive on-site 

screening of methylamphetamine seizures by attenuated total reflection 

Fourier transform infrared spectroscopy. APPLIED SPECTROSCOPY 

2008;62(6):640-648. 

63. Gosav S, Dinica R, Praisler M. Choosing between GC-FTIR and GC-MS 

spectra for an efficient intelligent identification of illicit amphetamines. 

JOURNAL OF MOLECULAR STRUCTURE 2008;887(1-3):269-278. 

64. Gosav S, Praisler M, Dorohoi DO. ANN expert system screening for illicit 

amphetamines using molecular descriptors. JOURNAL OF MOLECULAR 

STRUCTURE 2007;834:188-194. 

65. Inoue H, Kuwayama K, Iwata YT, Kanamori T, Tsujikawa K, Miyaguchi H. 

Simple and simultaneous detection of methamphetamine and dimethyl 

sulfone in crystalline methamphetamine seizures by fast gas 

chromatography. FORENSIC TOXICOLOGY 2008;26(1):19-22. 

66. Kim JY, In MG, Jung JC, Ko BJ. Method for simultaneously analyzing 

dimethylamphetamine and amphetamine derivatives by using liquid 

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67. Kubicz EJ, Allen L. Studies on methamphetamine detection with drugwipe 

analytical device, part 2. 2008 American Academy of Forensic Sciences 


68. Lamb PB, McElhinny CJ, Sninski T, Purdom H, Carroll FI, Lewin AH. High 

specific activity (+)-amphetamine and (+)-methamphetamine. JOURNAL 

OF LABELLED COMPOUNDS & RADIOPHARMACEUTICALS 

2009;52(11-12):457-462. 

69. Martinez FS, Roesch DM, Jacobs JL. Isolation of methamphetamine from 

1-(1’,4’-cyclohexadienyl)-2-methylaminopropane using potassium 

permanganate. JOURNAL OF THE CLANDESTINE LABORATORY 

INVESTIGATING CHEMISTS ASSOCIATION 2008;18(1):18. 

70. Martinez, FS, Roesch DM, and Jacobs JL. Isolation of methamphetamine 

from 1-(1’,4’-cyclohexadienyl)-2-methylaminopropane. MICROGRAM 

JOURNAL 2008;6(1-2):46. 

71. Matchett CC, Pietrucha TL. Investigation of mothballs as a counterfeit for 

“ice” methamphetamine. JOURNAL OF THE CLANDESTINE 


2008;19(2):10. 

72. Matsushita T, Takatsu M, Yoshida Y, Moriyasu M. Development of new 

on-column chiral derivatization reagent for gas chromatographic 

separation of optical isomeric amphetamine and methamphetamine. 

BUNSEKI KAGAKU 2007;56(12):1089-1095. 

73. McKetin R. Methamphetamine precursor regulation: Are we controlling or 

diverting the drug problem? ADDICTION 2008;103(4):521-523. 

74. Nevescanin M, Stevic SB, Petrovic S, Vajs V. Analysis of amphetamines 

illegally produced in Serbia. JOURNAL OF THE SERBIAN CHEMICAL 

SOCIETY 2008;73(7):691-701. 

75. Pavlova V, Petrovska-Jovanovic S. Simultaneous determination of 

amphetamine, methamphetamine, and caffeine in seized tablets by highperformance 

liquid chromatography. ACTA CHROMATOGRAPHICA 

2007;18:157. 

76. Romero GM, Chianella I, Piletska EV, Karim K, Turner APF, Piletsky SA. 

Development of a piezoelectric sensor for the detection of 

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2009;134(8):1565-1570. 

77. Sachs SB, Woo F. A detailed mechanistic fragmentation analysis of 

methamphetamine and select regioisomers by GC/MS. JOURNAL OF 

FORENSIC SCIENCES 2007;52(2):308-319. 

78. Sanderson R. Identification of N-methylbenzlamine hydrochloride, Nethylbenzylamine 

hydrochloride, and N-isopropylbenzylamine 

hydrochloride. MICROGRAM JOURNAL 2008;6(1-2):36. 

79. Shahdousti P, Aghai-Nohammadi M, Alizadeh N. Spectrophotometric 

study of the charge-transfer and ion-pair complexation of 

methamphetamine with some acceptors. SPECTROCHIMICA ACTA 

PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY 

2008;69(4):1195-1200. 

745

80. Smith JP, Martin A, Sammons DL, Striley C, Biagini R, Quinn J, Cope R, 

Snawder JE. Measurement of methamphetamine on surfaces using 

surface plasmon resonance. TOXICOLOGY MECHANISMS AND 

METHODS 2009;19(6-7):416-421. 

81. Steiner RR. A rapid technique for the confirmation of iodine and red 

phosphorus using direct analysis in real time and accurate mass 

spectrometry. MICROGRAM JOURNAL 2010;7(1):3-6. 

82. Sugita R, Sasagawa K, Suzuki S. Illegal route estimation of the seized 

illicit drug, methamphetamine, by the comparison of striation marks on 

plastic packaging films. JOURNAL OF FORENSIC SCIENCES 

2009;54(6):1341-1348. 

83. Tsujikawa K, Kuwayama K, Miyaquchi H, Kanamori T, Iwata YT, Yoshida 

T, Inoue H. Development of an on-site screening system for 

amphetamine-type stimulant tablets with a portable attenuated total 

reflection Fourier transform infrared spectrometer. ANALYTICA CHIMICA 

ACTA 2008;608(1):95-103. 

84. Wang H, Jia J, Cao J, Wang Y. Simultaneous determination of 

methamphetamine, caffeine and ketamine by GC/MS. SHANXI YIKE 

DAXUE XUEBAO 2007;38(11):1013-1016,1023. 

85. Watanabe-Galloway S, Ryan S, Hansen K, Hullsiek B, Muli V, Malone AC. 

Effects of methamphetamine abuse beyond individual users. JOURNAL 

OF PSYCHOACTIVE DRUGS 2009;41(3):241-248. 

86. Wei T, Hao H, Liu J, Zhu J, Yu, Z. Sensitive biosensor chip for recognizing 

methamphetamine, and method for manufacturing said biosensor chip. 

APPLICATION: CN 2010-118987 11 Mar 2009. 

87. Yohannan M. Detection of phenethylamine, amphetamine, and tryptamine 

imine by-products from an acetone extraction. Mid-Atlantic Association of 

Forensic Scientists Annual Meeting: 2009. 

88. Zaitsu K, Katagi M, Kamata H, Kamata T, Shima N, Miki A, Iwamura T, 

Tsuchihashi H. Discrimination and identification of the six aromatic 

positional isomers of trimethoxyamphetamine (TMA) by gas 

chromatography-mass spectrometry (GC-MS). JOURNAL OF MASS 

SPECTROMETRY 2008;43(4):528-534. 

89. Zhang LL, Chen Y, Lin M, Fan GR, Zhao WQ, Wu YT. Fast CE 

determination of d-amphetamine and diphenhydramine in quick-acting 

anti-motion capsules. CHROMATOGRAPHIA 2007;65(5-6):305. 

1.3 Barbiturates (including Quaaludes): 

90. Kuila DK, Lahiri SC. Study of hydrogen bonding interactions of 

methaqualone with alcohols, naphthols and nitrophenols. JOURNAL OF 

THE INDIAN CHEMICAL SOCIETY 2008;85(8):830-835. 

91. Pilau EJ, Silva RGC, Jardim ICFS, Augusto F. Molecularly imprinted solgel 

silica for solid phase extraction of phenobarbital. JOURNAL OF THE 

BRAZILIAN CHEMICAL SOCIETY 2008;19(6):1136-43. 

746

92. Ray R, Sharma JD, Limaye SN. Variations in the physico-chemical 

parameters of some N-methyl substituted barbiturate derivatives. ASIAN 

JOURNAL OF CHEMISTRY 2007;19(5):3382. 

93. Shabir GA, Bradshaw TK, Arain SA, Shar GQ. A new validated method for 

the simultaneous determination of a series of eight barbiturates by RP- 

HPLC. JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED 

TECHNOLOGIES 2010;33(1):61-71. 

1.4 Benzodiazepines: 

94. Acikkol M, Mercan S, Karadayi S. Simultaneous determination of 

benzodiazepines and ketamine from alcoholic and nonalcoholic beverages 

by GC-MS in drug facilitated crimes (vol 70, pg 1295, 2009). 

CHROMATOGRAPHIA 2009;70(11-12):1769-1770. 

95. Acikkol M, Mercan S, Karadayi S. Simultaneous determination of 

benzodiazepines and ketamine from alcoholic and nonalcoholic beverages 

by GC-MS in drug facilitated crimes. CHROMATOGRAPHIA 2009;70(7- 

8):1295-1298. 

96. Bishop SC, Lerch M, McCord BR. Detection of nitrated benzodiazepines 

by indirect laser-induced fluorescence detection on a microfluidic device. 

JOURNAL OF CHROMATOGRAPHY A 2007;1154(1-2):481-484. 

97. Brettell TA, Asselin EL. Presumptive and confirmatory identification of 1,2triazolo-benzodiazepines. 

2008 American Academy of Forensic Sciences 


98. Daundkar BB, Malve MK, Krishnamurthy R. A specific chromogenic 

reagent for detection of diazepam among other benzodiazepines from 

biological and nonbiological samples after HPTLC. JOURNAL OF 

PLANAR CHROMATOGRAPHY—MODERN TLC 2008;21(4):249-250. 

99. Du H, Ge H, Yu L, Wang X. Fast detection of clonazepam mixed illegally 

in traditional Chinese medicine by TLCS. ZHONGGUO YAOSHI 

2007;10(9):891. 

100. Guthery B, Bassindale A, Pillinger CT, Morgan GH. The detection of 

various opiates and benzodiazepines by comprehensive two-dimensional 

gas chromatography/time-of-flight mass spectrometry. RAPID 

COMMUNICATIONS IN MASS SPECTROMETRY 2009;23(3):340-348. 

101. Honeychurch KC. Determination of flunitrazepam and nitrazepam in 

beverage samples by liquid chromatography with dual electrode detection 

using a carbon fibre veil electrode. JOURNAL OF SOLID STATE 

ELECTROCHEMISTRY 2008;12(10):1317-1324. 

102. Ishida T, Kudo K, Hayashida M, Ikeda N. Rapid and quantitative screening 

method for 43 benzodiazepines and their metabolites, zolpidem and 

zopiclone in human plasma by liquid chromatography/mass spectrometry 

with a small particle column. JOURNAL OF CHROMATOGRAPHY, B: 

ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE 

SCIENCES 2009;877(25):2652-2657. 

747

103. Janowska E, Adamowicz P, Chudzikiewicz E, Lechowicz W. Clonazepam- 

A drug use for medical and criminal purposes. Z ZAGADNIEN NAUK 

SADOWYCH 2007;71:297-302. 

104. Koba M, Koba K, Baczek T. Determination of oxazepam in pharmaceutical 

formulation by HPTLC UV-densitometric and uv-derivative 

spectrophotometry methods. ANALYTICAL LETTERS 2009;42(12):1831- 

1843. 

105. McGuire ND, Honeychurch KC, Hart, JP. The electrochemical behavior of 

nitrazepam at a screen-printed carbon electroge and its determination in 

beverages by adsorptive stripping voltammetry. ELECTROANALYSIS 

2009;21(19):2165-2170. 

106. Sarwar M, Taylor S, Majeed I. A specific screening color test for 

diazepam. MICROGRAM JOURNAL 2008;6(3-4):63. 

107. Webb R, Doble P, Dawson M. A rapid CZE method for the analysis of 

benzodiazepines in spiked beverages. ELECTROPHORESIS 

2007;28:3553-3565. 

1.5 Dimethoxyphenethylamines: 

108. da Costa JL, Wang AY, Micke GA, Maldaner AO, Romano RL, Martins 

HA, Negrini O, Tavares MFM. Chemical identification of 2,5-dimethoxy-4bromoamphetamine 

(DOB). FORENSIC SCIENCE INTERNATIONAL 

2007;173:130-136. 

109. Franck MC, Meneghini LZ, Rossato LG, Limberger RP, Froehlich PE. 

Development and validation of an LC-UV method for quantitation of 4bromo-2,5-dimethoxyamphetamine 

(DOB), 4-bromo-2,5dimethoxyphenetylamine 

(2C-B), methylphenidate, fenproporex and 

amfepramone. CHROMATOGRAPHIA 2009;69(Suppl. 2):143-148. 

110. Luiz da Costa J, Wang AY, Micke GA, Maldaner AO, Romano RL, 

Martins-Junior HA, Negrini O, Tavares MFM. Chemical identification of 

2,5-dimethoxy-4-bromoamphetamine (DOB). FORENSIC SCIENCE 

INTERNATIONAL 2007;173(2-3):130. 

111. Maher HM, Awad T, DeRuiter J, Clark CR. GC-MS and GC-IRD studies on 

dimethoxyamphetamines (DMA): Regioisomers related to 2,5-DMA. 

FORENSIC SCIENCE INTERNATIONAL 2009;192(1-3):115-125. 

112. Suzuki J, Takahashi M, Nagashima M, Seto T, Mori K, Ogino S. 

Identification of 2,4,5-trichloro-3,6-dimethoxyphenethylamine detected in 

illegal drugs by the chemical synthesis. TOKYO-TO KENKO ANZEN 

KENKYU SENTA KENKYU NENPO 2009;59:85-89. 

113. Trachsel D, Nichols DE, Kidd S, Hadorn M, Baumberger F. 4-Arylsubstituted 

2,5-dimethoxyphenethylamines: Synthesis and serotonin 5- 

HT2A receptor affinities. CHEMISTRY & BIODIVERSITY 2009;6(5):692- 

704. 

748

114. Xu YZ, Lin HR, Lua AC, Chen C. Determination of 

dimethoxyphenethylamine derivatives in urine by deuterium labeled 

internal standards. JOURNAL OF THE SERBIAN CHEMICAL SOCIETY 

2008;73(12):1223-1233. 

115. Xu YZ, Lin HR, Lua CA, Chen CP. Synthesis of deuterium labeled (+/-)- 

2,5-dimethoxyamphetamine (DMA), (+/-)-4-bromo-2,5dimethoxyamphetamine 

(DOB), and (+/-)-4-methoxyamphetamine (PMA) 

as internal standards for quantitative analyses by GC-MS. JOURNAL OF 

THE CHINESE CHEMICAL SOCIETY 2007;54(2):493-502. 

1.6 Chlordiazepoxide: 

116. Bakal RL, Manwar JV, Sahare AY, Bhajipale NS, Manikrao AM. 

Spectrophotometric estimation of amitriptyline HCl and chlordiazepoxide in 

pharmaceutical dosage form. INDIAN JOURNAL OF PHARMACEUTICAL 

EDUCATION AND RESEARCH 2008;42(1):23-26. 

117. Jouyban A, Shokri J, Barzegar-Jalali M, Hassanzadeh D, Acree 

WE, Ghafourian T, Nokhodchi A. Solubility of chlordiazepoxide, diazepam, 

and lorazepam in ethanol plus water mixtures at 303.2 K. JOURNAL OF 

CHEMICAL AND ENGINEERING DATA 2009;54(7):2142-2145. 

118. Ozdemir A, Keskin CS. Quantification of chlordiazepoxide and pipenzolate 

bromide in tablets and comparison with chromatographic results. ASIAN 

JOURNAL OF CHEMISTRY 2009;21(2):1159-1170. 

119. Patel S, Patel NJ, Patel SA. Simultaneous spectrophotometric estimation 

of imipramine hydrochloride and chlordiazepoxide in tablets. INDIAN 

JOURNAL OF PHARMACEUTICAL SCIENCES 2009;71(4):468-U198. 

120. Patel S, Patel NJ. Spectrophotometric and chromatographic simultaneous 

estimation of amitriptyline hydrochloride and chlordiazepoxide in tablet 

dosage forms. INDIAN JOURNAL OF PHARMACEUTICAL SCIENCES 

2009;71(4):472-U203. 

121. Patel SK, Patel NJ. Simultaneous RP-HPLC estimation of trifluoperazine 

hydrochloride and chlordiazepoxide in tablet dosage forms. INDIAN 

JOURNAL OF PHARMACEUTICAL SCIENCES 2009;71(5):545-547. 

122. Sarrafi AHM, Khodakarami Z, Karkeabadi M. Simultaneous 

spectrophotometric determination of amitriptyline hydrochloride and 

chlordiazepoxide in pharmaceutical tablets by multivariate calibration 

method. E-JOURNAL OF CHEMISTRY 2009;6:S111-S116. 

1.7 Clenbuterol: 

123. Jiao F, Chen X, Hu Y, Wang Z. Enantiomeric separation of racemic 

clenbuterol by capillary zone electrophoresis. JOURNAL OF THE 

IRANIAN CHEMICAL SOCIETY 2008;5(4):553-558. 

749

124. Mostafa GAE Hefnawy MM El-Majed A. Separation and determination of 

clenbuterol by HPLC using a vancomycin chiral stationary phase. 

JOURNAL OF AOAC INTERNATIONAL 2009;92(3):824-829. 

125. Parr MK, Koehler K, Geyer H, Guddat S, Schanzer W. Clenbuterol 

marketed as dietary supplement. BIOMEDICAL CHROMATOGRAPHY 

2008;22(3):298-300. 

1.8 Cocaine: 

126. Aehle E, Drager B. Tropane alkaloid analysis by chromatographic and 

electrophoretic techniques: An update. JOURNAL OF 

CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE 

BIOMEDICAL AND LIFE SCIENCES 2010;878(17-18):1391-1406. 

127. Anonymous. Washington, DC, USA, proves to be cocaine capital of the 

world. TrAC-TRENDS IN ANALYTICAL CHEMISTRY 2009;28(9):V-VI. 

128. Bonilla DA, Penuela LF, Sierra N, Diaz JE, Rojas JH. Development and 

validation of an analytical methodology for cocaine hydrochloride 

determination in a synthetic polymer by ultraviolet spectrometry. VITAE 

2008;15(1):103-12. 

129. Borges F, Cordeiro M, Mosquera RA, Natalia DS,Rincon DA. Theoretical 

study of cocaine and ecgonine methyl ester in gas phase and in aqueous 

solution. CHEMICAL PHYSICS LETTERS 2009;467(4-6):249-254. 

130. Boudreau DK, Casale JF. An in-depth study of the peruvian base llavada 

(“washed base”) technique for purification of crude cocaine base. 

MICROGRAM JOURNAL 2008;6(3-4):72. 

131. Brackney M, Baumbach J, Ewers C, Martinez AL, Hagan J, Czuchlewski 

D, Foucar K, Fekrazad MH, Seifert SA, Dimple D, Nolte KB, Buchanan JA, 

Lavonas Nelson C, Wood RW, Duchin JS, VanEenwyk J, Reuter N, Ta 

ML, Vagi S. Agranulocytosis associated with cocaine use-four states, 

March 2008-November 2009 (Reprinted from MMWR, vol 58, pg 1381- 

1385, 2009). JAMA-JOURNAL OF THE AMERICAN MEDICAL 

ASSOCIATION 2010;303(6):504. 

132. Bradley A, Millington A. Agricultural land-use trajectories in a cocaine 

source region: Chapare, Bolivia. LAND-CHANGE SCIENCE IN THE 

TROPICS: CHANGING AGRICULTURAL LANDSCAPES 2008;231-250. 

133. Casale JF, Boudreau DK, Jones LM. Tropane ethyl esters in illicit cocaine: 

Isolation, detection, and determination of new manufacturing by-products 

from the clandestine purification of crude cocaine base with ethanol. 

JOURNAL OF FORENSIC SCIENCES 2008;53(3):661-667. 

134. Casale JF, Corbeil EM, Hays PA. Identification of levamisole impurities 

found in illicit cocaine exhibits. MICROGRAM JOURNAL 2008;6(3-4):82. 

135. Casale J, Lydon J. Apparent effects of glyphosate on alkaloid production 

in coca plants grown in Colombia. JOURNAL OF FORENSIC SCIENCES 

2007;52(3):573-578. 

136. Casale JF, Nguyen MC. N-Acetylbenzocaine: Formation via 

transacetylation of benzocaine and acetylsalicylic acid in a cocaine exhibit. 

MICROGRAM JOURNAL 2010;7(1):7-11. 

750

137. Casale JF, Orlando PM, Colley VL, Hays PA. Identification of diltiazem 

impurities/artifacts during the analyses of illicit cocaine exhibits containing 

diltiazem. MICROGRAM JOURNAL 2008;6(3-4):90. 

138. Casale JF, Hays PA, Toske SG, Berrier AL. Four new illicit cocaine 

impurities from the oxidation of crude cocaine base: Formation and 

characterization of the diastereomeric 2,3-dihydroxy-3phenylpropionylecgonine 

methyl esters from cis- and transcinnamoylcocaine. 

JOURNAL OF FORENSIC SCIENCES 

2007;52(4):860-866. 

139. Cecinato A, Balducci C, Nervegna G. Occurrence of cocaine in the air of 

the world's cities: An emerging problem? A new tool to investigate the 

social incidence of drugs? SCIENCE OF THE TOTAL ENVIRONMENT 

2009;407(5):1683-1690. 

140. Corbeil EM, Casale JF. Determination of cocaine in various south 

american commercial coca products. MICROGRAM JOURNAL 2008;6(3- 

4):109. 

141. Czuchlewski DR, Brackney M, Ewers C, Manna J, Fekrazad MH, Martinez 

A, Nolte KB, Hjelle B, Rabinowitz I, Curtis BR, McFarland JG, Baumbach 

J, Foucar K. Clinicopathologic features of agranulocytosis in the setting of 

levamisole-tainted cocaine. AMERICAN JOURNAL OF CLINICAL 

PATHOLOGY 2010;133(3):466-472. 

142. Da Silva MJ, dos Anjos EV, Honorator RS, Pimentel MF, Paim AP. 

Spectrophotometric cocaine determination in a biphasic medium 

employing flow-batch sequential injection analysis. ANALYTICA CHIMICA 

ACTA 2008;629(1-2):98-103. 

143. Davalos LM, Bejarano AC, Correa HL. Disabusing cocaine: Pervasive 

myths and enduring realities of a globalised commodity. INTERNATIONAL 

JOURNAL OF DRUG POLICY 2009;20(5):381-386. 

144. Eliasson C, Macleod NA, Matousek P. Non-invasive detection of cocaine 

dissolved in beverages using displaced Raman spectroscopy. 

ANALYTICA CHIMICA ACTA 2008;607(1):50-53. 

145. Gheorghe A, van Nuijs A, Pecceu B, Bervoets L, Jorens PG, Blust R, 

Neels H, Covaci A. Analysis of cocaine and its principal metabolites in 

waste and surface water using solid phase extraction and liquid 

chromatography-ion trap tandem mass spectrometry. ANALYTICAL & 

BIOANALYTICAL CHEMISTRY 2008;391(4):1309-19. 

146. Gostic T, Klemenc S, Stefane B. A study of the thermal decomposition of 

adulterated cocaine samples under optimized aerobic pyrolytic conditions. 


147. Gostic T, Klemenc S. Evidence on unusual way of cocaine smuggling: 

Cocaine-polymethyl methacrylate (PMMA) solid solution - study of 

clandestine laboratory samples. FORENSIC SCIENCE INTERNATIONAL 

2007;169(2-3):210-219. 

148. He JL, Wu ZS, Zhou H, Wang HQ, Jiang JH, Shen GL, Yu RQ. 

Fluorescence aptameric sensor for strand displacement amplification 

detection of cocaine. ANALYTICAL CHEMISTRY 2010;82(4):1358-1364. 

751

149. Hewitt AJ, Solomon KR, Marshall EJP. Spray droplet size, drift potential, 

and risks to nontarget organisms from aerially applied glyphosate for coca 

control in Colombia. JOURNAL OF TOXICOLOGY AND 

ENVIRONMENTAL HEALTH-PART A-CURRENT ISSUES 2009;72(15- 

16):921-929. 

150. Janowska E. Peruvian foodstuffs with cocaine. Z ZAGADNIEN NAUK 

SADOWYCH 2008;75:276-281. 

151. Jones LM, Boudreau DK, Casale JF. “Crack” cocaine: A study of stability 

over time and temperature. MICROGRAM JOURNAL 2008;6(3-4):114. 

152. Kinzie E. Levamisole found in patients using cocaine. ANNALS OF 

EMERGENCY MEDICINE 2009;53(4):546-547. 

153. Knowles L, Buxton JA, Skuridina N, Achebe I, LeGatt D, Fan SH, Zhu NY, 

Talbot J. Levamisole tainted cocaine causing severe neutropenia in 

Alberta and British Columbia. HARM REDUCTION JOURNAL 2009;6. 

154. Kolomiets YN, Pervukhin VV. Effect of UV irradiation on detection of 

cocaine hydrochloride and crack vapors by IMIS and API-MS methods. 

TALANTA 2009;78(2):542-547. 

155. Mari F, Politi L, Biggeri A, Accetta G, Trignano C, Di Padua M, Bertol E. 

Cocaine and heroin in waste water plants: A 1-year study in the city of 

Florence, Italy. FORENSIC SCIENCE INTERNATIONAL 2009;189(1- 

3):88-92. 

156. Marshall EJP, Solomon KR, Carrasquilla G. Coca (Erythroxylum coca) 

control is affected by glyphosate formulations and adjuvants. JOURNAL 

OF TOXICOLOGY AND ENVIRONMENTAL HEALTH-PART A-CURRENT 

ISSUES 2009;72(15-16):930-936. 

157. Matthews AR, Oladapo O, Morgan BW. A cocaine death with concomitant 

adulterant-induced toxicity. CLINICAL TOXICOLOGY 2009;47(7):290. 

158. McGill JW, Dixon CA, Ritter D, Sides JD. Discovery of an interesting 

temperature effect on the sensitivity of cobalt thiocyanate test for cocaine. 


159. Moore JM, Casale JF. The discoloration of illicit drug samples. 


160. Oiye EN, de Figueiredo NB, de Andrade JF, Tristao HM, de Oliveira MF. 

Voltammetric determination of cocaine in confiscated samples using 

a cobalt hexacyanoferrate film-modified electrode. FORENSIC SCIENCE 

INTERNATIONAL 2009;192(1-3):94-97. 

161. Pedersoli S, Lombardi L, Hoehr NF, Rittner R. Assignments of H-1 and C- 

13 NMR spectral data for benzoylecgonine, a cocaine metabolite. 

SPECTROSCOPY LETTERS 2008;41(3):101-103. 

162. Pesaresi M. Textural analysis of coca plantations using remotely sensed 

data with resolution of 1 metre. INTERNATIONAL JOURNAL OF 

REMOTE SENSING 2008;29(23):6985-7002. 

752

163. Pistos C, Karampela S, Papoutsis I, Athanaselis S, Spiliopoulou Ch, 

Maravelias C. Investigation of the identification point system adaptation in 

cocaine, benzoylecgonine and ecgonine methyl ester using a single 

quadrupole mass spectrometer. RAPID COMMUNICATIONS IN MASS 

SPECTROMETRY 2009;23(23):3772-3780. 

164. Qaseem A, Shekelle P, Owens DK. Agranulocytosis after consumption of 

cocaine adulterated with levamisole. ANNALS OF INTERNAL MEDICINE 

2009;150(4):287-289. 

165. Raymon LP, Isenschmid DS. The possible role of levamisole in illicit 

cocaine preparations. JOURNAL OF ANALYTICAL TOXICOLOGY 

2009;33(9):620-622. 

166. Rincon DA, Cordeiro M, Mosquera RA, Borges F. Theoretical study of 

cocaine and ecgonine methyl ester in gas phase and in aqueous solution. 

CHEMICAL PHYSICS LETTERS 2009;467(4-6):249-254. 

167. Ropero-Miller JD, Stout PR, Bynum ND, Casale JF. Comparison of the 

novel direct analysis in real time time-of-flight mass spectrometry 

(AccuTOF-DART) and signature analysis for the identification of 

constituents of refined illicit cocaine. MICROGRAM JOURNAL 2007;5(1- 

4):34-40. 

168. Shiyahovsky B, Di L, Weizmann Y, Nowarski R, Kotler M, Willner I. 

Spotlighting of cocaine by an autonomous aptamer-based machine. 

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 

2007;129(13):3814. 

169. Smith RM, Casale JF. The mass spectrum of cocaine: deuterium labeling 

and MS/MS studies. MICROGRAM JOURNAL 2010;7(1):16-41. 

170. Storti CC, De Grauwe P. The cocaine and heroin markets in the era of 

globalisation and drug reduction policies. INTERNATIONAL JOURNAL OF 

DRUG POLICY 2009;20(6):488-496. 

171. Tadmor T, Vadasz Z, Polliack A, Attias D. Two cases of agranulocytosis 

associated with cocaine use: Is this phenomenon becoming more 

prevalent? EUROPEAN JOURNAL OF HAEMATOLOGY 2010;84(5):458- 

459. 

172. Wiens MO, Son WK, Ross C, Hayden M, Carleton B. Cocaine adulterant 

linked to neutropenia. CANADIAN MEDICAL ASSOCIATION JOURNAL 

2009;182(1):57-59. 

173. Zhang CY, Johnson LW. Single quantum-dot-based aptameric 

nanosensor for cocaine. ANALYTICAL CHEMISTRY 2009;81(8):3051- 

3055. 

1.9 Emerging Substances (including MDPV and “FLY” compounds) 

174. Andreasen MF, Telving R, Birkler RI, Schumacher B, Johannsen M. A 

fatal poisoning involving Bromo-Dragonfly. FORENSIC SCIENCE 


753

175. Bozenko JS, Yohannan JC. The characterization of 3,4methylenedioxypyrovalerone 

(MDPV). MICROGRAM JOURNAL 

2010;7(1):12-15. 

176. Funada M. Uncontrolled newly-abused drugs (law-evading drugs). 

NIPPON YAKURIGAKU ZASSHI 2007;130(5):433-435. 

177. Kenney J. Not So Legal, “Legal Highs”. JOURNAL OF THE 

CLANDESTINE LABORATORY INVESTIGATING CHEMISTS 


178. Kikura-Hanajiri R, Kawamura M, Uchiyama N, Ogata J, Kamakura H, 

Saisho K, Goda Y. Analytical data of designated substances (Shitei- 

Yakubutsu) controlled by the pharmaceutical affairs law in Japan, part I: 

GC-MS and LC-MS (vol 128, pg 971, 2008). YAKUGAKU ZASSHI- 

JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN 

2008;128(12):1857-1857. 

179. Kikura-Hanajiri R, Kawamura M, Uchiyama N, Ogata J, Kamakura H, 

Saisho K, Goda Y. Analytical data of designated substances (Shitei- 

Yakubutsu) controlled by the pharmaceutical affairs law in Japan, part I: 

GC-MS and LC-MS. YAKUGAKU ZASSHI-JOURNAL OF THE 

PHARMACEUTICAL SOCIETY OF JAPAN 2008;128(6):971-979. 

180. Kikura-Hanajiri R, Maruyama T, Miyashita A, Goda Y. Chemical and DNA 

analyses for the products of a psychoactive plant, Voacanga africana. 

YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY 

OF JAPAN 2009;129(8):975-982. 

181. Masahiko F. Uncontrolled newly-abused drugs (law-evading drugs). 

FOLIA PHARMACOLOGICA JAPONICA 2007;130(5):433-435. 

182. Nagashima J, Seto T, Takahashi M, Suzuki J, Mori K, Ogino S. Analyses 

of uncontrolled drugs purchased from April 2007 to March 2008 and newly 

found compounds. TOKYO-TO KENKO ANZEN KENKYU SENTA 

KENKYU NENPO 2009;59:71-77. 

183. Piggee C. Investigating a not-so-natural-high. ANALYTICAL CHEMISTRY 

2009. 

184. Reed EC, Kiddon GS. The characterization of three FLY compounds. 

MICROGRAM JOURNAL 2007;5(1-4):26-33. 

185. Sauer C, Peters FT, Haas C, Meyer MR, Fritschi G, Maurer HH. New 

designer drug alpha-pyrrolidinovalerophenone (PVP): Studies on 

its metabolism and toxicological detection in rat urine using 

gas chromatographic/mass spectrometric techniques. JOURNAL OF 

MASS SPECTROMETRY 2009;44(6):952-964. 

186. Thevis M, Thomas A, Kohler M, Beuck S, Schaenzer W. Emerging drugs: 

Mechanism of action, mass spectrometry and doping control analysis. 

JOURNAL OF MASS SPECTROMETRY 2009;44(4):442-460. 

187. Uchiyama N, Kawamura M, Kamakura H, Kikura-Hanajiri R, Goda Y. 

Analytical data of designated substances (shitei-yakubutsu) controlled by 

the pharmaceutical affairs law in Japan, part II: Color test and TLC. 


OF JAPAN 2008;128(6):981-987. 

754

188. Westphal F, Junge T, Roesner P, Soennichsen F, Schuster F. Mass and 

NMR spectroscopic characterization of 3,4-methylenedioxypyrovalerone: 

A designer drug with alpha-pyrrolidinophenone structure. FORENSIC 


189. Wood DM, Button J, Lidder S, Ovaska H, Ramsey J, Holt DW, Dargan PI. 

Detection of the novel recreational drug diphenyl-2-pyrrolidinemethanol 

(D2PM) sold 'legally' in combination with glaucine. CLINICAL 

TOXICOLOGY 2008;46(5):193-. 

190. Yohannan J. Driving under the influence, but under the influence of what? 

An introduction to MDPV. Mid-Atlantic Association of Forensic Scientists 

Annual Meeting: 2009. 

191. Zaitsu K, Katagi M, Kamata H, Nakanishi K, Shima N, Kamata T, Nishioka 

H, Miki A, Tatsuno M, Tsuchihashi H. Simultaneous analysis of six novel 

hallucinogenic (tetrahydrobenzodifuranyl)aminoalkanes (FLYs) and 

(benzodifuranyl)aminoalkanes (DragonFLYs) by GC-MS, LC-MS, and LC- 

MS-MS. FORENSIC TOXICOLOGY 2010;28(1):9-18. 

192. Zhang ZY, An LY, Hu WX, Xiang YH. 3D-QSAR study of hallucinogenic 

phenylalkylamines by using CoMFA approach. JOURNAL OF 

COMPUTER-AIDED MOLECULAR DESIGN 2007;21(4):145. 

1.10 Ergot Alkaloids (see also LSD): 

193. Crews C, Anderson WAC, Rees G, Krska R. Ergot alkaloids in some ryebased 

UK cereal products. FOOD ADDITIVES & CONTAMINANTS PART 

B-SURVEILLANCE 2009;2(1):79-85. 

194. Krska R, Berthiller F, Schuhmacher R, Nielsen KF, Crews C. 

Determination of ergot alkaloids: Purity and stability assessment 

of standards and optimization of extraction conditions for cereal samples. 

JOURNAL OF AOAC INTERNATIONAL 2008;91(6):1363-1371. 

195. Krska R, Crews C. Significance, chemistry and determination of ergot 

alkaloids: A review. FOOD ADDITIVES & CONTAMINANTS 

2008;25(6):722-731. 

196. Krska R, Stubbings G, Macarthur R, Crews C. Simultaneous 

determination of six major ergot alkaloids and their epimers in cereals and 

foodstuffs by LC-MS-MS. ANALYTICAL & BIOANALYTICAL CHEMISTRY 

2008;391(2):563-576. 

197. Muller C, Kemmlein S, Klaffke H, Krauthause W, Preiss-Weigert 

A, Wittkowski R. A basic tool for risk assessment: A new method for the 

analysis of ergot alkaloids in rye and selected rye products. MOLECULAR 

NUTRITION & FOOD RESEARCH 2009;53(4):500-507. 

755

1.11 Fentanyl(s): 

198. Cicero TJ, Inciardi JA, Surratt H. Trends in the use and abuse of branded 

and generic extended release oxycodone and fentanyl products in the 

United States. DRUG AND ALCOHOL DEPENDENCE 2007;91:115-120. 

199. Denton JS, Donoghue ER, McReynolds J, Kalelkar MB. An epidemic of 

illicit fentanyl deaths in Cook County, Illinois: September 2005 through 

April 2007. JOURNAL OF FORENSIC SCIENCES 2008;53(2):452-454. 

200. Gupta PK, Manral L, Ganesan K, Dubey DK. Use of single-drop 

microextraction for determination of fentanyl in water samples. 

ANALYTICAL & BIOANALYTICAL CHEMISTRY 2007;388(3):579-583. 

201. Higashikawa Y, Suzuki S. Studies on 1-(2-phenethyl)-4-(Npropionylanilino)piperidine 

(fentanyl) and its related compounds: Novel 

metabolites in rat urine following injection of alpha-methylfentanyl, one of 

the most abused typical designer drugs. JOURNAL OF HEALTH 

SCIENCE 2008;54(6):629-637. 

202. Higashikawa Y, Suzuki S. Studies on 1-(2-phenethyl)-4-(Npropionylanilino) 

piperidine (fentanyl) and it related compounds. VI. 

Structure-analgesic activity relationship for fentanyl, methyl-substituted 

fentanyls and other analogues. FORENSIC TOXICOLOGY 2008;26(1):1- 

5. 

203. Lurie IS, Iio R. Use of multiple-reaction monitoring ratios for identifying 

incompletely resolved fentanyl homologs and analogs via ultra-highpressure 

liquid chromatography-tandem mass spectrometry. JOURNAL 

OF CHROMATOGRAPHY A 2009;1216(9):1515-1519. 

204. Ojanpera I, Gergov M, Liiv M, Riikoja A, Vuori E. An epidemic of fatal 3methylfentanyl 

poisoning in Estonia. INTERNATIONAL JOURNAL OF 

LEGAL MEDICINE 2008;122(5):395-400. 

205. VanNimmen NFJ, Veulemans HAF. Validated GC-MS analysis for the 

determination of residual fentanyl in applied Durogesic® reservoir and 

Durogesic® D-Trans® Matrix transdermal fentanyl patches. JOURNAL OF 

CHROMATOGRAPHY B - ANALYTICAL TECHNOLOGIES IN THE 

BIOMEDICAL AND LIFE SCIENCES 2007;846(1-2):264. 

206. Vuckovic S, Prostran M, Ivanovic M, Dosen-Micovic L, Todorovic Z, Nesic 

Z, Stojanovic R, Divac N, Mikovic Z. Fentanyl analogs: Structure-activityrelationship 

study. CURRENT MEDICINAL CHEMISTRY 

2009;16(19):2468-2474. 

207. Wang Y, Dai H, Wu X, Chen H, Xu L, Chen Y, Chen G. 

Electrochemiluminescence determination of fentanyl citrate with a 

novel glassy carbon paste electrode. LUMINESCENCE 2008;23(2):99-99. 

208. Wong SC, Curtis JA, Wingert WE. Concurrent detection of heroin, 

fentanyl, and xylazine in seven drug-related deaths reported from the 

Philadelphia Medical Examiner's Office. JOURNAL OF FORENSIC 

SCIENCES 2008;53(2):495-498. 

756

209. Yamamoto K, Kojima M, Iguchi H, Ohta T. Measurement of amount of 

fentanyl remaining in used patches: Investigation of clinical factors 

affecting the remaining amounts in 4 patients. YAKUGAKU ZASSHI- 


2008;128(6):959-964. 

1.12 Flos Daturae: 

210. Li JG, Chun Y, Ju HX. Simultaneous electrochemiluminescence detection 

of anisodamine, atropine, and scopolamine in Flos daturae by capillary 

electrophoresis using beta-Cyclodextrin as additive. ELECTROANALYSIS 

2007;19(15):1569-1574. 

1.13 Fluoxetine (Prozac): 

211. Patel RB, Patel MR, Shankar MB, Bhatt KK. Simultaneous determination 

of alprazolam and fluoxetine hydrochloride in tablet formulations by highperformance 

column liquid chromatography and high-performance thinlayer 

chromatography. JOURNAL OF AOAC INTERNATIONAL 

2009;92(4):1082-1088. 

212. Patel S, Patel NJ. Simultaneous RP-HPLC and HPTLC estimation of 

fluoxetine hydrochloride and olanzapine in tablet dosage forms. INDIAN 

JOURNAL OF PHARMACEUTICAL SCIENCES 2009;71(4):477-480. 

1.14 Heroin: 

213. Agg KM, Barnett NW, Lewis SW, Pearson JR. Preliminary investigations 

into tris(2,2'-bipyridyl) ruthenium (III) as a chemiluminescent reagent for 

the detection of 3,6-diacetylmorphine (heroin) on surfaces. JOURNAL OF 


214. Agg KM, Craddock AF, Bos R, Francis PS, Lewis SW, Barnett NW. A 

rapid test for heroin (3,6-diacetylmorphine) based on 

two chemiluminescence reactions. JOURNAL OF FORENSIC SCIENCES 

2007;52(3):759-759. 

215. Buryakov IA, Baldin MN. Comparison of negative and positive modes of 

ion-mobility increment spectrometry in the detection of heroin vapors. 

JOURNAL OF ANALYTICAL CHEMISTRY 2008;63(8):787-791. 

216. Casale JF, Toske SG, Hays PA. Chlorinated opium alkaloid derivatives 

produced by the use of aqueous sodium hypochlorite during the 

clandestine manufacture of heroin. JOURNAL OF FORENSIC SCIENCES 

2009;54(2):359-364. 

217. Ciccarone D, Unick GJ, Kraus A. Impact of South American heroin on the 

US heroin market 1993-2004. INTERNATIONAL JOURNAL OF DRUG 

POLICY 2009;20(5):392-401. 

757

218. Dussy FE, Berchtold C, Briellmann TA, Lang C, Steiger R, Bovens M. 

Validation of an ion mobility spectrometry (IMS) method for the detection 

of heroin and cocaine on incriminated material. FORENSIC SCIENCE 


219. Gheorghe M, Balalau D, Ilie M, Baconi DL, Ciobanu AM. Component 

analysis of illicit heroin samples by GC-MS method. FARMACIA 

2008;56(5):577-582. 

220. Hill LA, Lenehan CE, Francis PS, Adcock JL, Gange ME, Pfeffer FM, 

Barnett NW. A screening test for heroin based on sequential injection 

analysis with dual-reagent chemiluminescence detection. TALANTA 

2008;76(3):674-679. 

221. Jiggens J. Australian heroin seizures and the causes of the 2001 heroin 

shortage. INTERNATIONAL JOURNAL OF DRUG POLICY 

2008;19(4):273-278. 

222. Kerr SC, Casale JF. The use of dipropionylmorphine as a structurallyrelated 

internal standard for gas chromatographic quantitation of heroin. 


223. Kleinschmidt K, Feng S, Goto C, Sobotka A, Liebbe J. Difficulty in the 

forensic laboratory assessment of "Cheese" - A new form of heroin. 

CLINICAL TOXICOLOGY 2007;45:224. 

224. Liu M, Song C, Qiao J, Wang Y. Component and purity of retail heroin and 

concentration ratio of morphine to codeine in urine of heroin abusers. 

ZHONGGUO YAOWU YILAIXING ZAZHI 2007;16(5):386-9. 

225. Moros J, Galipienso N, Vilches R, Garrigues S, de la Guardia M. 

Nondestructive direct determination of heroin in seized illicit street drugs 

by diffuse reflectance near-infrared spectroscopy. ANALYTICAL 

CHEMISTRY 2008;80(19):7257-7265. 

226. Moros J, Kuligowski J, Quintas G, Garrigues S, de la Guardia M. New cutoff 

criterion for uninformative variable elimination in multivariate calibration 

of near-infrared spectra for the determination of heroin in illicit street 

drugs. ANALYTICA CHIMICA ACTA 2008;630(2):150-160. 

227. Risser D, Uhl A, Oberndorfer F, Honigschnabl S, Stichenwirth M, Hirz 

R, Sebald D. Is there a relationship between street heroin purity and drugrelated 

emergencies and/or drug-related deaths? An analysis from 

Vienna, Austria. JOURNAL OF FORENSIC SCIENCES 2007;52(5):1171- 

1176. 

228. Rudakov TN, Hayes PA, Flexman JH. Optimized NQR pulse technique for 

the effective detection of heroin base. SOLID STATE NUCLEAR 

MAGNETIC RESONANCE 2008;33(3):31-35. 

229. Subhan F, Khan N, Sewell RDE. Adulterant profile of illicit street heroin 

and reduction of its precipitated physical dependence withdrawal 

syndrome by extracts of St. John's Wort (Hypericum perforatum). 

PHYTOTHERAPY RESEARCH 2009;23(4):564-571. 

230. Toprak S, Cetin I. Heroin overdose deaths and heroin purity between 1990 

and 2000 in istanbul, turkey. JOURNAL OF FORENSIC SCIENCES 

2009;54(5):1185-1188. 

758

231. Wodak A. What caused the recent reduction in heroin supply in Australia? 


232. Wu G-P, Xiang B-R. A new method for fast and nondestructive analysis of 

heroin, 6-acetylmorphine, and codeine in drug by near infrared 

spectroscopy. FENXI HUAXUE 2007;35(4):552. 

233. Yang S, Liu X, Qiao J. Analysis of diluents in retail heroin seized in Beijing 

in 2005. ZHONGGUO YAOWU YILAIXING ZAZHI 2007;16(4):276-80. 

234. Zhang ZX, Yan B, Liu KL, Bo T, Liao YP, Liu HW. Fragmentation 

pathways of heroin-related alkaloids revealed by ion trap and quadrupole 

time-of-flight tandem mass spectrometry. RAPID COMMUNICATIONS IN 


235. Zhang ZX, Yan B, Liu KL, Liao YP, Liu HW. CE-MS analysis of heroin and 

its basic impurities using a charged polymer-protected gold nanoparticlecoated 

capillary. ELECTROPHORESIS 2009;30(2):379-387. 

1.15 gamma-Hydroxybutyric Acid (GHB), gamma-butyrolactone (GBL) and 

1,4-butanediol (BD): 

236. Bennett MJ. Detection of GHB in various drink matrices via AccuTOF- 

DART. 2008 American Academy of Forensic Sciences Annual Meeting. 

237. Bennett MJ, Steiner RR. Detection of gamma-hydroxybutyric acid in 

various drink matrices via AccuTOF-DART. JOURNAL OF FORENSIC 

SCIENCES 2009;54(2):370-375. 

238. Brewster VL, Edwards HGM, Hargreaves MD, Munshi T. Identification of 

the date-rape drug GHB and its precursor GBL by Raman spectroscopy. 

DRUG TESTING AND ANALYSIS 2009;1(1-2):25-31. 

239. Carter LP, Pardi D, Gorsline J, Griffiths RR. Illicit gamma-hydroxybutyrate 

(GHB) and pharmaceutical sodium oxybate [Xyrem (R)]: Differences in 

characteristics and misuse. DRUG AND ALCOHOL DEPENDENCE 

2009;104(1-2):1-10. 

240. Dempsey DA, Wiegand TJ, Havel C, Benowitz NL, Haller C. Butanediol 

(BD) Conversion to gamma-hydroxybutyrate, (GHB), is markedly reduced 

by fomepizole (4-MP), an alcohol dehydrogenase blocker. CLINICAL 

TOXICOLOGY 2009;47(7):113. 

241. Elie MP, Baron MG, Birkett JW. Enhancement of microcrystalline 

identification of gamma-hydroxybutyrate. JOURNAL OF FORENSIC 

SCIENCES 2008;53(1):147-150. 

242. Henck C, Nally L. GC-MS analysis gamma-hydroxybutyric acid analogs: A 

forensic chemistry experiment. JOURNAL OF CHEMICAL EDUCATION 

2007;84:1813-1815. 

243. Majid MA. Identification of gamma-hydroxybutyrate (GHB), gammabutyrolactone 

(GBL) and 1,4-butanediol using trimethyl derivatization. Mid- 

Atlantic Association of Forensic Scientists Annual Meeting: 2009. 

244. Marinetti L. History and pharmacology of γ-hydroxybutyric acid. 

CHROMATOGRAPHIC METHODS IN CLINICAL CHEMISTRY AND 

TOXICOLOGY 2007:197. 

759

245. Meng PJ. Analysis of gamma-hydroxybutyric acid using gas 

chromatography/mass spectrometry after pentafluorobenzyl derivatization. 

FENXI HUAXUE 2008;36(1):61-65. 

246. Mercer JW, Oldfield LS, Hoffman KN, Shakleya DM, Bell SC. Comparative 

analysis of gamma-hydroxybutyrate and gamma-hydroxyvalerate using 

GC/MS and HPLC. JOURNAL OF FORENSIC SCIENCES 

2007;52(2):383-388. 

247. Michalik KA, Brettell TA. The detection of gamma-hydroxybutyric acid 

through the use of a rapid colorimetric test. 2008 American Academy of 

Forensic Sciences Annual Meeting. 

248. Saudan C, Augsburger M, Mangin P, Saugy M. Carbon isotopic ratio 

analysis by gas chromatography/combustion/isotope ratio mass 

spectrometry for the detection of gamma-hydroxybutyric acid (GHB) 

administration to humans. RAPID COMMUNICATIONS IN MASS 

SPECTROMETRY 2007;21(24):3956-3962. 

1.16 Human Growth Hormone (HGH): 

249. Hancock WS, Jiang H, Karger BL,Wu SL. Mass spectrometric analysis of 

innovator, counterfeit, and follow-on recombinant human growth hormone. 

BIOTECHNOLOGY PROGRESS 2009;25(1):207-218. 

1.17 Ketamine: 

250. Pieri M, Castiglia L, Miraglia N, Guadagni R, Malorni L, Sannolo 

N, Acampora A, Della Casa E. Study of the fragmentation pattern of 

ketamine-heptafluorobutyramide by gas chromatography/electron 

ionization mass spectrometry. RAPID COMMUNICATIONS IN MASS 


251. Wang G, Shen J, Jia Y. Vibrational spectra of ketamine hydrochloride and 

3,4-methylenedioxymethamphetamine in terahertz range. JOURNAL OF 

APPLIED PHYSICS 2007;102(1):013106/1. 

1.18 Khat: 

252. Balint EE, Falkay G, Balint GA. Khat - a controversial plant. WIENER 

KLINISCHE WOCHENSCHRIFT 2009;121(19-20):604-614. 

253. Chappell JS, Lee MM. Cathinone preservation in khat evidence via drying. 

FORENSIC SCIENCE INTERNATIONAL 2010;195(1-3):108-120. 

254. Krizevski R, Dudai N, Bar E, Dessow I, Ravid U, Lewinsohn E. 

Quantitative stereoisomeric determination of phenylpropylamino alkaloids 

in khat (Catha edulis Forsk.) using chiral GC-MS. ISRAEL JOURNAL OF 

PLANT SCIENCES 2008;56(3):207-213. 

760

255. Krizevski R, Dudai N, Bar E, Lewinsohn E. Developmental patterns of 

phenylpropylamino alkaloids accumulation in khat (Catha edulis Forsk.). 

JOURNAL OF ETHNOPHARMACOLOGY 2007;114(3):432-438. 

256. Tefera TL. Supply response, local reality and livelihood sustainability: The 

policy dilemma of khat (Catha edulis) production in eastern Ethiopia. 

INTERNATIONAL JOURNAL OF AGRICULTURAL SUSTAINABILITY 

2009;7(3):176-188. 

1.19 Kratom (Also mitragynine): 

257. Assanangkornchai S, Muekthong A, Sam-Angsri N, Pattanasattayawong 

U. The use of Mitragynine speciosa ("Kratom"), an addictive plant, in 

Thailand. SUBSTANCE USE & MISUSE 2007;42(14):2145-2157. 

258. Chittrakarn S, Keawpradub N, Sawangjaroen K, Kansenalak 

S, Janchawee B. Mitragynine and methanol extract of kratom leaf 

(Mitragyna speciosa Korth.) inhibited compound nerve action potential. 

DRUG METABOLISM REVIEWS 2009;41(205). 

259. Kikura-Hanajiri R, Kawamura M, Maruyama T, Kitajima M, Takayama H, 

Goda Y. Simultaneous analysis of mitragynine, 7-hydroxymitragynine, and 

other alkaloids in the psychotropic plant "kratom" (Mitragyna speciosa) by 

LC-ESI-MS. FORENSIC TOXICOLOGY 2009;27(2):67-74. 

260. Leon F, Habib E, Adkins JE, Furr EB, McCurdy CR, Cutler SJ. 

Phytochemical characterization of the leaves of mitragyna speciosa grown 

in USA. NATURAL PRODUCT COMMUNICATIONS 2009;4(7):907-910. 

261. Liu HN, McCurdy CR, Doerksen RJ. Computational study on the 

conformations of mitragynine and mitragynaline. JOURNAL OF 

MOLECULAR STRUCTURE-THEOCHEM 2010;945(1-3):57-63. 

262. Maruyama T, Kawamura M, Kikura-Hanajiri R, Takayama H, Goda Y. The 

botanical origin of kratom (Mitragyna speciosa; Rubiaceae) available as 

abused drugs in the Japanese markets. JOURNAL OF NATURAL 

MEDICINES 2009;63(3):340-344. 

263. Phongprueksapattana S, Putalun W, Keawpradub N, Wungsintaweekul J. 

Mitragyna speciosa: Hairy root culture for triterpenoid production and high 

yield of mitragynine by regenerated plants. ZEITSCHRIFT FUR 

NATURFORSCHUNG SECTION C-A JOURNAL OF BIOSCIENCES 

2008;63(9-10):691-698. 

264. Roche KM, Hart K, Sangalli B, Lefberg J, Bayer M. Kratom: A case of a 

legal high. CLINICAL TOXICOLOGY 2008;46(7):41-. 

265. Sukrong S, Zhu S, Ruangrungsi N, Phadungcharoen T, Palanuvej C, 

Komatsu K. Molecular analysis of the genus Mitragyna existing in Thailand 

based on rDNA ITS sequences and its application to identify a narcotic 

species: Mitragyna speciosa. BIOLOGICAL & PHARMACEUTICAL 

BULLETIN 2007;30(7):1284-1288. 

761

1.20 LSD: 

267. Inoue T, Yokoshima S, Fukuyama T. Synthetic studies toward (+)-Lysergic 

acid: Construction of the tetracyclic ergoline skeleton. HETEROCYCLES 

2009;79:373-378. 

268. Moldvai I, Gacs-Baitz E, Termesvari-Major E, Russo L, Papai I, Rissanen 

K, Szarics E, Kardos J, Szantay C. Dimerization of (+)-lysergic acid esters. 

HETEROCYCLES 2007;71(5):1075-1094. 

1.21 Marijuana and Related Cannabinoids: 

269. Aarons JN. Laboratory and field experiments used for the determination of 

odor signature chemicals in marijuana. 2008 American Academy of 


270. Ahmed SA, Ross SA, Slade D, Radwan MM, Khan IA, Elsohly MA. 

Structure determination and absolute configuration of cannabichromanone 

derivatives from high potency Cannabis sativa. TETRAHEDRON 

LETTERS 2008;49(42):6050-6053. 

271. Ahmed SA, Ross SA, Slade D, Radwan MM, Zulfiqar F, Matsumoto RR, 

Xu YT, Viard E, Speth RC, Karamyan VT, ElSohly MA. Cannabinoid ester 

constituents from high-potency Cannabis sativa. JOURNAL OF NATURAL 

PRODUCTS 2008;71(6):1119-1119. 

272. Ahmed SA, Ross SA, Slade D, Radwan MM, Zufiqar A, ElSohly MA. 

Isolation and characterization of new cannabis constituents from a 

high potency variety. PLANTA MEDICA 2008;74(3):P15. 

273. Ahmed SA, Ross SA, Slade D, Radwan MM, Zulfiqar F, ElSohly MA. 

Cannabinoid ester constituents from high-potency Cannabis sativa. 

JOURNAL OF NATURAL PRODUCTS 2008;71(4):536-542. 

274. Appendino G, Giana A, Gibbons S, Maffei M, Gnavi G, Grassi G, Sterner 

O. A polar cannabinoid from Cannabis sativa var. Carma. NATURAL 

PRODUCT COMMUNICATIONS 2008;3(12):1977-1980. 

275. Appendino G, Gibbons S, Giana A, Pagani A, Grassi G, Stavri M, Smith 

E, Rahman M. Antibacterial cannabinoids from Cannabis sativa: A 

structure-activity study. JOURNAL OF NATURAL PRODUCTS 

2008;71(8):1427-1430. 

276. Balducci C, Nervegna G, Cecinato A. Evaluation of principal cannabinoids 

in airborne particulates. ANALYTICA CHIMICA ACTA 2009;641(1-2):89- 

94. 

277. Boleda MR, Galceran MT, Ventura F. Trace determination of cannabinoids 

and opiates in wastewater and surface waters by ultra-performance liquid 

chromatography-tandem mass spectrometry. JOURNAL OF 

CHROMATOGRAPHY A 2007;1175(1):38. 

762

278. Chou SL, Ling YC, Yang MH, Pai CY. Determination of delta(9)tetrahydrocannabinol 

in indoor air as an indicator of marijuana cigarette 

smoking using adsorbent sampling and in-injector thermal desorption gas 

chromatography-mass spectrometry. ANALYTICA CHIMICA ACTA 

2007;598(1):103-109. 

279. De Backer B, Debrus B, Lebrun P, Theunis L Dubois N, Decock L 

Verstraete A, Hubert P, Charlier C. Innovative development and validation 

of an HPLC/DAD method for the qualitative and quantitative determination 

of major cannabinoids in cannabis plant material. JOURNAL OF 

CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE 

BIOMEDICAL AND LIFE SCIENCES 2009;877(32):4115-4124. 

280. de Meijer EPM, Hammond KM, Micheler M. The inheritance of chemical 

phenotype in Cannabis sativa L. (III): Variation in cannabichromene 

proportion. EUPHYTICA 2009;165(2):293-311. 

281. de Meijer EPM, Hammond KM, Sutton A. The inheritance of chemical 

phenotype in Cannabis sativa L. (IV): Cannabinoid-free plants. 

EUPHYTICA 2009;168(1):95-112. 

282. de Oliveira GL, Voloch MH, Sztulman GB, Neto ON, Yonamine M. 

Cannabinoid contents in cannabis products seized in Sao Paulo, 

Brazil, 2006-2007. FORENSIC TOXICOLOGY 2008;26(1):31-35. 

283. Fischedick JT, Glas R, Hazekamp A, Verpoorte R. A Qualitative and 

quantitative HPTLC densitometry method for the analysis of cannabinoids 

in Cannabis sativa L. PHYTOCHEMICAL ANALYSIS 2009;20(5):421-426. 

284. Flemming T, Muntendam R, Steup C, Kayser O. Chemistry and biological 

activity of tetrahydrocannabinol and its derivatives. TOPICS IN 

HETEROCYCLIC CHEMISTRY 2007;10(Bioactive Heterocycles IV):1-42. 

285. Florian NM, Parada F, Garzon WF. Study Of cannabinoids content in 

marihuana samples (Cannabis Sativa L.) cultivated in several regions of 

Colombia. VITAE-REVISTA DE LA FACULTAD DE QUIMICA 

FARMACEUTICA 2009;16(2):237-244. 

286. Fucci N. Analysis of fatty acids in marijuana (Cannabis sativa leaf). 

MICROGRAM JOURNAL 2007;5(1-4):20-6. 

287. Garcia A, Borchardt D, Chang CA, Marsella MJ. Thermal isomerization of 

cannabinoid analogues. JOURNAL OF THE AMERICAN CHEMICAL 

SOCIETY 2009;131(46):16640-16641. 

288. Hakki EE; Kayis SA, Pinarkara E, Sag A. Inter simple sequence repeats 

separate efficiently hemp from marijuana (Cannabis sativa L.) 

ELECTRONIC JOURNAL OF BIOTECHNOLOGY 2007;10(4):None. 

289. Holler JM, Bosy TZ, Dunkley CS, Levine B, Past MR, Jacobs A. 

Tetrahydrocannabinol content of commercially available hemp products. 

JOURNAL OF ANALYTICAL TOXICOLOGY 2008;32(6):428-432. 

290. Holler JM, Smith ML, Paul SN, Past MR, Paul BD. Isomerization of delta- 

9-THC to delta-8-THC when tested as trifluoroacetyl-, 

pentafluoropropionyl-, or heptafluorobutyryl-derivatives. JOURNAL OF 


763

291. Howard C, Gilmore S, Robertson J, Peakall R. A Cannabis STR genotype 

database for Australian seizures: forensic applications and limitations. 


292. Howard C, Gilmore S, Robertson J, Peakall R. Developmental validation 

of a cannabis sativa STIR multiplex system for forensic analysis. 


293. Knight G, Hansen S. An experimental indoor hydroponic cannabis growing 

set-up, using the screen of green (ScrOG) method. JOURNAL OF THE 



294. Lee JS, Park YH, Rhee JS, Jeong JI, Lim MA, Chung HS. Planting 

conditions of Korean cannabis derived from stable isotope ratio and 

tetrahydrocannabinol contents. YAKHAK HOECHI 2008;52(3):172-175. 

295. McLaren J, Swift W, Dillon P, Allsop S. Cannabis potency and 

contamination: A review of the literature. ADDICTION 2008;103(7):1100- 

1109. 

296. Mehmedic Z, Slade D, Denham H, Chandra S, Stanford D, Khan IA, 

ElSohly MA. Indoor and outdoor cultivation of cannabis: Is there an effect 

on the chemical composition of the volatile oil? PLANTA MEDICA 

2008;74(3):P17. 

297. Mendoza MA, Mills DK, Lata H, Chandra S, ElSohly MA, Almirall JR. 

Genetic individualization of Cannabis sativa by a short tandem repeat 

multiplex system. ANALYTICAL & BIOANALYTICAL CHEMISTRY 

2009;393(2):719-726. 

298. Ogata J, Kikura-Hanajiri R, Yoshimatsu K, Kiuchi F, Goda Y. Detection 

method for the ability of hemp (Cannabis sativa L.) seed germination by 

the use of 2,3,5-triphenyl-2H-tetrazolium chloride (TTC). YAKUGAKU 

ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN 

2008;128(11):1707-1711. 

299. Pacifico D, Miselli F, Carboni A, Moschella A, Mandolino G. Time course 

of cannabinoid accumulation and chemotype development during the 

growth of Cannabis sativa L. EUPHYTICA 2008;160(2):231-240. 

300. Peltzer K, Ramlagan S. Cannabis use trends in South Africa. SOUTH 

AFRICAN JOURNAL OF PSYCHIATRY 2007;13(4):126-131. 

301. Pinarkara E, Kayis SA, Hakki EE, Sag A. RAPD analysis of seized 

marijuana (Cannabis sativa L.) in Turkey. ELECTRONIC JOURNAL OF 

BIOTECHNOLOGY 2009;12(1). 

302. Potter DJ, Clark P, Brown MB. Potency of delta(9)-THC and other 

cannabinoids in cannabis in England in 2005: Implications for 

psychoactivity and pharmacology. JOURNAL OF FORENSIC SCIENCES 

2008;53(1):90-94. 

303. Radwan MM, ElSohly MA, Slade D, Ahmed SA, Khan IA, Ross SA. 

Biologically active cannabinoids from high-potency Cannabis sativa. 

JOURNAL OF NATURAL PRODUCTS 2009;72(5):906-911. 

764

304. Radwan MM, ElSohly MA, Slade D, Ahmed SA, Wilson L, Al-Alfy AT, 

Ross SA. Non-cannabinoid constituents from a high potency Cannabis 

sativa variety. PLANTA MEDICA 2008;74(9):1069-1069. 

305. Radwan MM, ElSohly MA, Slade D, Ahmed SA, Wilson L, El-Alfy AT, 

Khan IA, Ross SA. Non-cannabinoid constituents from a high potency 

Cannabis sativa variety. PHYTOCHEMISTRY 2008;69(14):2627-2633. 

306. Radwan MM, Ross SA, Slade D, Ahmed SA, Zulfiqar F, ElSohly MA. 

Isolation and characterization of new cannabis constituents from 

high potency variety. PLANTA MEDICA 2008;74(3):267-272. 

307. Ricordel I, Wenzek M. Cannabis and safety of work. Evolution of its 

detection within the controls of narcotics since 2004 to the SNCF. 

ANNALS PHARMACEUTIQUES FRANCAISES 2008;66(4):255-260. 

308. Stanaszek R, Zuba D. A comparison of developed and validated 

chromatographic methods (HPLC, GC-MS) for determination of delta-9tetrahydrocannabinol 

and delta-9-tetrahydrocannabinolic acid in hemp. Z 

ZAGADNIEN NAUK SADOWYCH 2007;71:313-322. 

309. Van der Kooy F, Pomahacova B, Verpoorte R. Cannabis smoke 

condensate II: Influence of tobacco on tetrahydrocannabinol levels. 

INHALATION TOXICOLOGY 2009;21(2):87-90. 

310. Ware MA, St Arnaud-Trempe E. The abuse potential of the synthetic 

cannabinoid nabilone. ADDICTION 2010;105(3):494-503. 

311. Yoshimatsu K, Kitazawa T, Kawano N, Iida O, Kawahara N. 

Characteristics of Cannabis sativa L.: Seed Morphology, Germination and 

Growth Characteristics, and Distinction from Hibiscus cannabinus L. 


OF JAPAN 2010;130(2):237-246. 

1.22 Mescaline/Peyote: 

312. Carstairs SD, Cantrell FL. Crazy for Cacti: A retrospective review of 

peyote exposures. CLINICAL TOXICOLOGY 2009;47(7):57. 

313. Casado R, Uriarte I, Cavero RY, Calvo MI. LC-PAD determination of 

mescaline in cactus "Peyote" (Lophophora williamsii). 

CHROMATOGRAPHIA 2008;67(7-8):665-667. 

314. MacLean H. Rock crystals and peyote dreams: Explorations in the Huichol 

universe. JOURNAL OF ANTHROPOLOGICAL RESEARCH 

2008;64(3):448-449. 

315. Mann J, Mann J. Peyote and amphetamines. TURN ON AND TUNE IN: 

PSYCHEDELICS, NARCOTICS AND EUPHORIANTS 2009;108-118,138- 

139. 

316. Nakajima J, Aragane M, Hamano T, Shioda H, Yoshizawa M, Suzuki Y, 

Kitagawa S, Yasuda I Mori K, Ogino S. Analysis of mescaline in botanical 

uncontrolled drugs prepared from cactus. TOKYO-TO KENKO ANZEN 

KENYU SENTA KENKYU NENPO 2009;59:91-95. 

765

1.23 Methadone: 

317. Snozek CLH, Bjergum MW, Langman LJ. Gas Chromatography-Mass 

Spectrometry method for the determination of methadone and 2ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine 

(EDDP). CLINICAL 

APPLICATIONS OF MASS SPECTROMETRY: METHODS AND 

PROTOCOLS 2010;351-358. 

1.24 Methcathinone (Also methcathinone-related compounds): 

318. Archer RP. Fluoromethcathinone, a new substance of abuse. FORENSIC 


319. Belal T, Awad T, DeRuiter J, Clark CR. GC-IRD methods for the 

identification of isomeric ethoxyphenethylamines and 

methoxymethcathinones. FORENSIC SCIENCE INTERNATIONAL 

2009;184(1-3):54-63. 

320. Camilleri A, Johnston MR, Brennan M, Davis S, Caldicott DGE. Chemical 

analysis of four capsules containing the controlled substance analogues 4methylmethcathinone, 

2-fluoromethamphetamine, alphaphthalimidopropiophenone 

and N-ethylcathinone. FORENSIC SCIENCE 


321. Combs MR. Analytical profile of 4-methylmethcathinone. JOURNAL OF 

THE CLANDESTINE LABORATORY INVESTIGATING CHEMISTS 


322. Dickson AJ, Vorce SP, Levine B, Past MR. Multiple-drug toxicity caused 

by the coadministration of 4-methylmethcathinone (Mephedrone) and 

heroin. JOURNAL OF ANALYTICAL TOXICOLOGY 2010;34(3):162-168. 

323. Dyer C. Mephedrone is an amphetamine "by another name," drug adviser 

tells MPs. BRITISH MEDICAL JOURNAL 2010;340. 

324. Fleming N. The truth about mephedrone. NEW SCIENTIST 

2010;206(2757):42-45. 

325. Kmietowicz Z. Home secretary bans mephedrone after taking advice from 

depleted council. BRITISH MEDICAL JOURNAL 2010;340. 

326. Meyer MR, Wilhelm J, Peters FT, Maurer HH. Beta-keto amphetamines: 

studies on the metabolism of the designer drug mephedrone and 

toxicological detection of mephedrone, butylone, and methylone in urine 

using gas chromatography-mass spectrometry. ANALYTICAL & 

BIOANALYTICAL CHEMISTRY 2010;397(3):1225-1233. 

327. Morris K. UK places generic ban on mephedrone drug family. LANCET 

2010;375(9723):1333-1334. 

328. Winstock AR, Marsden J, Mitcheson L. What should be done about 

mephedrone? BRITISH MEDICAL JOURNAL 2010;340. 

329. Zuba D. Medicines containing ephedrine and pseudoephedrine as a 

source of methcathinone. Z ZAGADNIEN NAUK SADOWYCH 

2007;71:323-333. 

766

1.25 Methylenedioxyamphetamines and Related Compounds: 

330. Awad T, Belal T, DeRuiter J, Clark CR. GC-IRD studies on regioisomeric 

ring substituted methoxy methyl phenylacetones related to 3,4methylenedioxyphenylacetone. 

FORENSIC SCIENCE INTERNATIONAL 

2010;194(1-3):39-48. 

331. Belal T, Awad T, DeRuiter J, Clark CR. GC-MS studies on acylated 

derivatives of 3-methoxy-4-methyl- and 4-methoxy-3-methylphenethylamines: 

Regioisomers related to 3,4-MDMA. FORENSIC 

SCIENCE INTERNATIONAL 2008;178(1):61-82. 

332. Boushehri SVS, Tamimi M, Kebriaeezadeh A. Quantitative determination 

of 3,4-methylenedioxymethamphetamine by thin-layer chromatography in 

ecstasy illicit pills in Tehran. TOXICOLOGY MECHANISMS AND 

METHODS 2009;19(9):565-569. 

333. Bruhn JG, El-Seedi HR, Stephanson N, Beck O, Shulgin AT. Ecstasy 

analogues found in cacti. JOURNAL OF PSYCHOACTIVE DRUGS 

2008;40(2):219-222. 

334. Choodum A, Thavarungkul P, Kanatharana P, Smith NW. Ecstasy 

analysis by monolithic materials-capillary electrochromatography. 

ANALYTICAL SCIENCES 2009;25(4):517-522. 

335. Collins M, Heagney A, Cordaro F, Odgers D, Tarrant G, Stewart S. Methyl 

3-[3 ',4 '-(methylenedioxy)phenyl]-2-methyl glycidate: An ecstasy 

precursor seized in Sydney, Australia. JOURNAL OF FORENSIC 

SCIENCES 2007;52(4):898-903. 

336. da Costa JL, Pintao ER, Corrigliano CMC, Neto ON. Determination Of 3,4methylenedioxymethamphetamine 

(MDMA) In ecstasy tablets by high 

performance liquid chromatography with fluorescence detection (HPLC- 

FD). QUIMICA NOVA 2009;32(4):965-969. 

337. de Korompay A, Hill JC, Carter JF, NicDaeid N, Sleeman R. Supported 

liquid-liquid extraction of the active ingredient (3,4methylenedioxymethylamphetamine) 

from ecstasy tablets for 

isotopic analysis. JOURNAL OF CHROMATOGRAPHY A 2008;1178(1- 

2):1-8. 

338. Deluca P, Schifano F, Psychonaut 2002 Research Group. Searching the 

internet for drug-related web sites: Analysis of online available information 

on ecstasy (MDMA). AMERICAN JOURNAL ON ADDICTIONS 

2007;16(6):479-483. 

339. Giraudon I, Bello PY. Monitoring ecstasy content in France: Results from 

the national surveillance system 1999-2004. SUBSTANCE USE & 

MISUSE 2007;42(10):1567-78. 

340. Gura S, Guerra-Diaz P, Lai H, Almirall JR. Enhancement in sample 

collection for the detection of MDMA using a novel planar SPME (PSPME) 

device coupled to ion mobility spectrometry (IMS). DRUG TESTING AND 

ANALYSIS 2009;1(7-8):355-362. 

767

341. Hakey PM, Allis DG, Hudson MR, Korter TM. Density functional 

dependence in the theoretical analysis of the terahertz spectrum of the 

illicit drug MDMA (Ecstasy). IEEE SENSORS JOURNAL 2010;10(3):478- 

484. 

342. Inoue H, Hashimoto H, Watanabe S, Iwata YT, Kanamori T, Miyaguchi H, 

Tsujikawa K, Kuwayama K, Tachi N, Uetake N. Thermal desorption 

counter-flow introduction atmospheric pressure chemical ionization for 

direct mass spectrometry of ecstasy tablets. JOURNAL OF MASS 

SPECTROMETRY 2009;44(9):1300-1307. 

343. Kato N, Fujita S, Ohta H, Fukuba M, Toriba A, Hayakawa K. Thin layer 

chromatography/fluorescence detection of 3,4-methylenedioxymethamphetamine 

and related compounds. JOURNAL OF FORENSIC 

SCIENCES 2008;53(6):1367-1371. 

344. Kikura-Hanajiri R, Kawamura M, Miyajima A, Sunouchi M, Goda Y. 

Determination of a new designer drug, N-hydroxy-3,4methylenedioxymethamphetamine 

and its metabolites in rats using ultraperformance 

liquid chromatography-tandem mass spectrometry. 


345. Lasmar MC, Leite EMA. Development and validation of a gas 

chromatography method for the determination of ecstasy and 

amphetamine derivatives in tablets. REVISTA BRASILEIRA DE 

CIENCIAS FARMACEUTICAS 2007;43(2):223. 

346. Liu J, Decatur J, Proni G, Champeil E. Identification and quantitation 

of 3,4-methylenedioxy-N-methylamphetamine (MDMA, ecstasy) in human 

urine by H-1 NMR spectroscopy. Application to five cases of intoxication. 


347. Luiz da Costa J, Pintao ER, Corrigliano CM, Negrini Neto O. 

Determination of 3,4-methylenedioxymethamphetamine (MDMA) in Ectasy 

tablets by high performance liquid chromatography with fluorescence 

detection (HPLC-FD). QUIMICA NOVA 2009;32(4):965-969. 

348. Matsuda K, Fukuzawa T, Ishii Y, Yamada H. Color reaction of 3,4methylenedioxyamphetamines 

with chromotropic acid: Its improvement 

and application to the screening of seized tablets. FORENSIC 

TOXICOLOGY 2007;25(1):37. 

349. Milhazes N, Martins P, Uriarte E, Garrido J, Calheiros R, Marques MPM, 

Borges F. Electrochemical and spectroscopic characterization of 

amphetamine-like drugs: Application to the screening of 3,4methylenedioxymethamphetamine 

(MDMA) and its synthetic precursors. 

ANALYTICA CHIMICA ACTA 2007;596(2):231. 

350. Salouros H, Collins M, Tarrant G, George AV. N-Cyanomethyl-N-methyl-1- 

(3 ',4 '-methylenedioxyphenyl)-2-propylamine: An MDMA manufacturing 

by-product. JOURNAL OF FORENSIC SCIENCES 2008;53(5):1083-1091. 

351. Thigpen AL, DeRuiter J, Clark CR. GC-MS studies on the regioisomeric 

2,3-and 3,4-methylenedioxyphenethylamines related to MDEA, MDMMA, 

and MBDB. JOURNAL OF CHROMATOGRAPHIC SCIENCE 

2007;45(5):229-235. 

768

352. Tsujikawa K, Kuwayama K, Miyaguchi H, Kanamori T, Iwata YT, Inoue H. 

Degradation of N-hydroxy-3,4-methylenedioxymethamphetamine in 

aqueous solution and its prevention. FORENSIC SCIENCE 


353. Tsujikawa K, Kuwayama K, Miyaguchi H, Kanamori T, Iwata YT, Inoue H. 

Increase in split ratio enables detection of underivatized N-hydroxy-3,4methylenedioxymethamphetamine 

and N-hydroxy-3,4methylenedioxyamphetamine 

by capillary GC-MS. FORENSIC 

TOXICOLOGY 2010;28(1):55-57. 

354. Vogels N, Brunt TM, Rigter S, van Dijk P, Vervaeke H, Niesink RJM. 

Content of ecstasy in the Netherlands: 1993-2008. ADDICTION 

2009;104(12):2057-2066. 

355. Wang G, Shen J. Terahertz spectroscopic investigation of methylenedioxy 

amphetamine. PROCEEDINGS OF SPIE 2008;6840. 

356. Wu G-p, Xiang B-r. Nondestructive determination of MDMA and MA in 

ecstasy by near infrared spectroscopy. FENXI CESHI XUEBAO 

2007;26(5):698. 

357. Yu XJ, Li GP. Determination of MDA and MDMA by TFA derivatization- 

GC/MS/MS. GUANGPU SHIYANSHI 2009;26(4):1051-1053. 

358. Zaitsu K, Katagi M, Kamata HT, Miki A, Tsuchihashi H. Discrimination and 

identification of regioisomeric beta-keto analogues of 3,4methylenedioxyamphetamines 

by gas chromatography-mass 

spectrometry. FORENSIC TOXICOLOGY 2008;26(2):45-51. 

1.26 Morphine, Codeine, and Related Opium Alkaloids: 

359. Chaudhary V, Leisch H, Moudra A, Allen B, De Luca V, Cox DP, Hudlicky 

T. Biotransformations of morphine alkaloids by fungi: Ndemethylations, 

oxidations, and reductions. COLLECTION OF 

CZECHOSLOVAK CHEMICAL COMMUNICATIONS 2009;74(7-8):1179- 

1193. 

360. Feng SY, Chen WW, Huang W, Cheng M, Lin JQ, Li YZ, Chen R. Surfaceenhanced 

Raman spectroscopy of morphine in silver colloid. CHINESE 

OPTICS LETTERS 2009;7(11):1055-1057. 

361. Hansen SH. Sample preparation and separation techniques for 

bioanalysis of morphine and related substances. JOURNAL OF 

SEPARATION SCIENCE 2009;32(5-6):825-834. 

362. Li F, Song J, Gao D, Zhang Q, Han D, Niu L. Simple and rapid 

voltammetric determination of morphine at electrochemically pretreated 

glassy carbon electrodes. TALANTA 2009;79(3):845-850. 

363. Pejic N, Blagojevic S, Vukelic J, Kolar-Anic L, Anic S. Analyte pulse 

perturbation technique for the determination of O-6-acetylmorphine in 

seized street drug samples. BULLETIN OF THE CHEMICAL SOCIETY 

OF JAPAN 2007;80:1942-1948. 

769

364. Rezaei B, Khayamian T, Mokhtari A. Simultaneous determination of 

codeine and noscapine by flow-injection chemiluminescence method 

using N-PLS regression. JOURNAL OF PHARMACEUTICAL AND 

BIOMEDICAL ANALYSIS 2009;49(2):234-239. 

1.27 Opiate Alkaloids (Also Opioids): 

365. Berenyi S, Csutoras C, Sipos A. Recent developments in the chemistry of 

thebaine and its transformation products as pharmacological targets. 

CURRENT MEDICINAL CHEMISTRY 2009;16(25):3215-3242. 

366. Bladek J, Polak P, Jarzyna E, Jakubowska I. Analysis of opiates and 

cannabinols by TLC. BIULETYN WOJSKOWEJ AKADEMII 

TECHNICZNEJ 2007;56(3):257-267. 

367. Bogusz MJ. Opiods: Methods of forensic analysis. HANDBOOK OF 

ANALYTICAL SEPARATIONS 2008;6:3-72. 

368. Hamunen K, Paakkari P, Kalso E. Trends in opioid consumption in the 

Nordic countries 2002-2006. EUROPEAN JOURNAL OF PAIN 

2009;13(9):954-962. 

369. Hindson BJ, Francis PS, Purcell SD, Barnett NW. Determination of opiate 

alkaloids in process liquors using capillary electrophoresis. JOURNAL OF 

PHARMACEUTICAL AND BIOMEDICAL ANALYSIS 2007;43(3):1164. 

370. Jensen H, Kirby DA. Assessment of the smokeability of oxycodone hcl 80 

mg controlled-release tablets. JOURNAL OF THE CLANDESTINE 


2008;20(1):5. 

371. Kumar MG, Lin Senshang. Hydromorphone: Analytical methodologies for 

its determination. CURRENT ANALYTICAL CHEMISTRY 2008;4(2):11- 

126. 

372. Mann J, Mann J. Opiates from opium to heroin. TURN ON AND TUNE IN: 

PSYCHEDELICS, NARCOTICS AND EUPHORIANTS 2009;26-59,137. 

373. Turner NW, Cauchi M, Piletska EV, Preston C, Piletsky SA. Rapid 

qualitative and quantitative analysis of opiates in extract of poppy head via 

FTIR and chemometrics: Towards in-field sensors. BIOSENSORS & 

BIOELECTRONICS 2009;24(11):3322-3328. 

1.28 Opium (and Opium Poppies): 

374. Apuya NR, Park JH, Zhang L, Ahyow M, Davidow P, Van Fleet J, Rarang 

JC, Hippley M, Johnson TW, Yoo HD, Trieu A, Krueger S, Wu CY, Lu YP, 

Flavell RB, Bobzin SC. Enhancement of alkaloid production in opium and 

California poppy by transactivation using heterologous regulatory factors. 

PLANT BIOTECHNOLOGY JOURNAL 2008;6(2):160-175. 

375. Chakrabarti DK, Singh OP. Techniques to reproduce primary and 

secondary symptoms of downy mildew disease in opium poppy (Papaver 

somniferum L.). ADVANCES IN PLANT SCIENCES 2008;21(1):65-66. 

770

376. Dittbrenner A, Mock HP, Borner A, Lohwasser U. Variability of alkaloid 

content in Papaver somniferum L. JOURNAL OF APPLIED BOTANY AND 

FOOD QUALITY-ANGEWANDTE BOTANIK 2009;82(2):103-107. 

377. Drea S, Hileman LC, De Martino G, Irish VF. Functional analyses of 

genetic pathways controlling petal specification in poppy. DEVELOPMENT 

2007;134:4157-4166. 

378. Dubey MK, Shasany AK, Dhawan OP, Shukla AK, Khanuja SPS. AFLP 

studies on downy-mildew-resistant and downy-mildew-susceptible 

genotypes of opium poppy. JOURNAL OF GENETICS 2010;89(1):9-19. 

379. Dubey MK, Dhawan OP, Khanuja SPS. Downy mildew resistance in 

opium poppy: Resistance sources, inheritance pattern, genetic variability 

and strategies for crop improvement. EUPHYTICA 2009;165(1):177-188. 

380. Dubey RB, Jain SK, Maloo SR. Combining ability and heterosis for latex 

yield, seed yield and other agronomic traits in opium poppy (Papaver 

somniferum L.). INDIAN JOURNAL OF GENETICS AND PLANT 

BREEDING 2007;67(4):392-395. 

381. Facchini PJ, De Luca V. Opium poppy and Madagascar periwinkle: Model 

non-model systems to investigate alkaloid biosynthesis in plants. PLANT 

JOURNAL 2008;54(4):763-784. 

382. Facchini PJ, Hagel JM, Liscombe DK, Loukanina N, MacLeod 

BP. Samanani N, Zulak KG. Opium poppy: Blueprint for an alkaloid 

factory. PHYTOCHEMISTRY REVIEWS 2007;6(1):97-124. 

383. Finlay IG, Mancroft L, Field F. Opium production in Afghanistan - Let's reexamine 

buying the Afghan poppies that are left. BRITISH MEDICAL 

JOURNAL 2008;336(7657):1325-1325. 

384. Foster RA, ONeill NR, Saunders JA. Potential biological control of opium 

poppy by fungal pathogens. PLANTA MEDICA 2008;74(3):P31. 

385. Francis PS, Adcock JL, Costin JW, Purcell SD, Pfeffer FM, Barnett NW. 

Chemiluminescence detection of opium poppy (Papaver somniferum) 

alkaloids. JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL 

ANALYSIS 2008;48(3):508-518. 

386. Gumuscu A, Arslan N, Sarihan EO. Evaluation of selected poppy 

(Papaver somniferum L.) lines by their morphine and other alkaloids 

contents. EUROPEAN FOOD RESEARCH AND TECHNOLOGY 

2008;226(5):1213-1220. 

387. Hagel JM, MacLeod BP, Facchini PJ. Opium poppy. BIOTECHNOLOGY 

IN AGRICULTURE AND FORESTRY 2007;169-187. 

388. Hagel JM, Weljie AM, Vogel HJ, Facchini PJ. Quantitative H-1 nuclear 

magnetic resonance metabolite profiling as a functional genomics platform 

to investigate alkaloid biosynthesis in opium poppy. PLANT 

PHYSIOLOGY 2008;147(4):1805-1821. 

389. Harvest T, Brown PH, Fist A, Gracie A, Gregory D, Koutoulis A. The latex 

capacity of opium poppy capsules is fixed early in capsule development 

and is not a major determinant in morphine yield. ANNALS OF APPLIED 

BIOLOGY 2009;154(2):251-258. 

771

390. Higashi Y, Smith TJ, Jez JM, Kutchan TM. Crystallization and preliminary 

X-ray diffraction analysis of salutaridine reductase from the opium poppy 

Papaver somniferum. ACTA CRYSTALLOGRAPHICA SECTION F- 

STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS 

2010;66:163-166. 

391. Khan R, Khan MMA, Singh M, Nasir S, Naeem M, Siddiqui 

MH, Mohammad F. Gibberellic acid and triacontanol can ameliorate the 

opium yield and morphine production in opium poppy (Papaver 

somniferum L.). ACTA AGRICULTURAE SCANDINAVICA SECTION B- 

SOIL AND PLANT SCIENCE 2007;57:307-312. 

392. Kutchan TM, Ounaroon A, Haase S, Frick S. O-Methyltransferases of 

tetrahydrobenzylisoquinoline alkaloid biosynthesis in Papaver 

somniferum. PN US 07514251 Donald Danforth Plant Science Center. 

Official Gazette of the United States Patent and Trademark Office Patents 

2009. 

393. Lin X, Wang J, Li L, Wang X, Lu H, Xie Z. Separation and determination of 

five major opium alkaloids with mixed mode of hydrophilic/cationexchange 

monolith by pressurized capillary electrochromatography. 

JOURNAL OF SEPARATION SCIENCE 2007;30(17):3011-3017. 

394. Lisson SN. Temperature and photoperiod effects on the growth and 

development of opium poppy (Papaver somniferum). AUSTRALIAN 

JOURNAL OF EXPERIMENTAL AGRICULTURE 2007;47(6):742-748. 

395. Montes-Borrego M, Ledesma FJM, Jimenez-Diaz RM, Landa BB. A 

Nested-polymerase chain reaction protocol for detection and population 

biology studies of peronospora arborescens,the downy mildew pathogen 

of opium poppy, using herbarium specimens and asymptomatic, fresh 

plant tissues. PHYTOPATHOLOGY 2009;99(1):73-81. 

396. Panicker S, Wojno HL, Ziska LH. Quantitation of the major alkaloids in 

opium from Papaver setigerum DC. MICROGRAM JOURNAL 2007;5(1- 

4):13-9. 

397. Pienkny S, Brandt W, Schmidt J, Jo RK, Ziegler J. Functional 

characterization of a novel benzylisoquinoline O-methyltransferase 

suggests its involvement in papaverine biosynthesis in opium poppy 

(Papaver somniferum L.). PLANT JOURNAL 2009;60(1):56-67. 

398. Reid RG, Durham DG, Boyle SP, Low AS, Wangboonskul J. 

Differentiation of opium and poppy straw using capillary electrophoresis 

and pattern recognition techniques. ANALYTICA CHIMICA ACTA 

2007;605(1):20-27. 

399. Remberg B, Sterrantino AF, Artner R, Janitsch C, Krenn L. Science in 

drug control: The alkaloid content of Afghan opium. CHEMISTRY & 

BIODIVERSITY 2008;5(9):1770-1779. 

400. Salehi P, Sonboli A, Zavareh AF, Sefidkon F, Dayeni M, Cheraghi B. 

Narcotic alkaloids of four Papaver species from Iran. ZEITSCHRIFT FÜR 

NATURFORSCHUNG C 2007;62(1-2):16. 

401. Shukla S, Singh SP, Singh HB, Pushpangadan P. High yielding multiple 

disease resistant/tolerant stable variety 'Madakini' of opium poppy. PN US 

772

07442854 AE Council of Scientific and Industrial Research. Official 

Gazette of the United States Patent and Trademark Office Patents 2008. 

402. Steinberg M, Taylor M. Marginalizing a vulnerable cultural and 

environmental landscape - Opium poppy production in highland 

Guatemala. MOUNTAIN RESEARCH AND DEVELOPMENT 

2007;27(4):318-321. 

403. Werb D, Kerr T, Montaner J, Wood E. The need for an evidence-based 

approach to controlling opium production in afghanistan. JOURNAL OF 

PUBLIC HEALTH POLICY 2008;29(4):440-448. 

404. Williams S. On islands, insularity, and opium poppies: Australia's secret 

pharmacy. ENVIRONMENT AND PLANNING D: SOCIETY AND SPACE 

2010;28(2):290-310. 

405. Yadav HK, Shukla S, Singh SP. Assessment of genotype x environment 

interactions for yield and morphine content in opium poppy (Papaver 

somniferum L.). ACTA AGRONOMICA HUNGARICA 2007;55(3):331-338. 

406. Yadav HK, Shukla S, Rastogi A, Singh SP. Assessment of diversity in new 

genetic stock of opium poppy (Papaver somniferum). INDIAN JOURNAL 

OF AGRICULTURAL SCIENCES 2007;77:537-539. 

407. Yadav HK, Shukla S, Rastogi A, Singh SP. Non-parametric measure of 

stability for yield and morphine content in opium poppy (Papaver 

somniferum). INDIAN JOURNAL OF AGRICULTURAL SCIENCES 

2007;77:596-599. 

408. Yadav HK, Shulkla S, Rastogi A, Singh MP, Singh SP. Genotype - 

Environmental interactions and phenotypic stability analyses of opium 

poppy (Papaver somniferum L.) for seed, opium and morphine content. 

INDIAN JOURNAL OF GENETICS AND PLANT BREEDING 

2007;67(2):166-168. 

409. Zein AL, Dakhil OO, Dawe LN, Georghiou PE. Enantioselective syntheses 

and X-ray structures of (S)- and (R)-N-norlaudanidine: Trace opium 

constituents. TETRAHEDRON LETTERS 2010;51(1):177-180. 

410. Zhang CJ, Cheng CG. Identification of Papaver somniferum L. and 

Papaver rhoeas using DSWT-FTIR-RBFNN. SPECTROSCOPY AND 

SPECTRAL ANALYSIS 2009;29(5):1255-1259. 

411. Ziegler J, Facchini PJ, Geissler R, Schmidt J, Ammer C, Kramell R, 

Voigtlander S, Gesell A, Pienkny S, Brandt W. Evolution of morphine 

biosynthesis in opium poppy. PHYTOCHEMISTRY 2009;70(15-16):1696- 

1707. 

412. Ziska L, Panicker S, Wojno H. Recent and projected increases in 

atmospheric carbon dioxide and the potential impacts on growth and 

alkaloid production in wild poppy (Papaver setigerum DC.). CLIMATIC 

CHANGE 2008;91(3-4):395-403. 

413. Zulak KG, Khan MF, Alcantara J, Schriemer DC, Facchini PJ. Plant 

Defense responses in opium poppy cell cultures revealed by liquid 

chromatography-tandem mass spectrometry proteomics. MOLECULAR & 

CELLULAR PROTEOMICS 2009;8(1):86-98. 

773

1.29 Phenethylamines (including mixtures of Amphetamines, 

Methylenedioxy-amphetamines, and Related Compounds): 

414. Awad T, DeRuiter J, Belal T, Clark CR. GC and mass spectral studies on 

acylated side chain regioisomers of 3-methoxy-4-methyl-phenethylamine 

and 4-methoxy-3-methyl-phenethylamine. JOURNAL OF 

CHROMATOGRAPHIC SCIENCE 2009;47(4):279-286. 

415. Awad T, DeRuiter J, Clark CR. GC-MS analysis of acylated derivatives of 

a series of side chain regioisomers of 2-methoxy-4-methylphenethylamines. 

JOURNAL OF CHROMATOGRAPHIC SCIENCE 

2008;46(5):375-380. 

416. Awad T, DeRuiter J, Clark CR. GC-MS analysis of ring and side chain 

regioisomers of ethoxyphenethylamines. JOURNAL OF 

CHROMATOGRAPHIC SCIENCE 2008;46(8):671-679. 

417. Kanai K, Takekawa K, Kumamoto T, Ishikawa T, Ohmori T. Simultaneous 

analysis of six phenethylamine-type designer drugs by TLC, LC-MS, and 

GC-MS. FORENSIC TOXICOLOGY 2008;26(1):6-12. 

418. Min JZ, Shimizu Y, Toyo'oka T, Inagaki S, Kikura-Hanajiri R, Goda Y. 

Simultaneous determination of 11 designated hallucinogenic 

phenethylamines by ultra-fast liquid chromatography with fluorescence 

detection. JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL 

TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES 

2008;873(2):187-194. 

419. Pellati F, Benvenuti S. Fast high-performance liquid chromatography 

analysis of phenethylamine alkaloids in citrus natural products on a 

pentafluorophenylpropyl stationary phase. JOURNAL OF 

CHROMATOGRAPHY A 2007;1165:58-66. 

420. Schultz DM, Prescher JA, Kidd S, Marona-Lewicka D, Nichols DE, Monte 

A. 'Hybrid' benzofuran-benzopyran congeners as rigid analogs 

of hallucinogenic phenethylamines. BIOORGANIC & MEDICINAL 

CHEMISTRY 2008;16(11):6242-6251. 

421. Thigpen AL, Awad T, DeRuiter J, Clark CR. GC-MS Studies on the 

Regioisomeric Methoxy-Methyl-Phenethylamines Related to MDEA, 

MDMMA, and MBDB. JOURNAL OF CHROMATOGRAPHIC SCIENCE 

2008;46(10):900-906. 

422. Westphal F, Roesner P, Junge T. Differentiation of regioisomeric ringsubstituted 

fluorophenethylamines with product ion spectrometry. 


1.30 Piperazines: 

423. Abdel-Hay KM, Awad T, DeRuiter J, Clark CR. Differentiation of 

methylenedioxybenzylpiperazines (MDBP) by GC-IRD and GC-MS. 


774

424. Elliott S, Smith C. Investigation of the first deaths in the United Kingdom 

involving the detection and quantitation of the piperazines BZP and 3- 

TFMPP. JOURNAL OF ANALYTICAL TOXICOLOGY 2008;32(2):172-177. 

425. Heegel RA, Trigg JJ. Different Approaches to the separation of 1benzylpiperazine 

and 1-[3-(trifluoromethyl)phenyl]-piperazine found in illicit 

ecstasy tablets. JOURNAL OF THE CLANDESTINE LABORATORY 


426. Johnstone AC, Lea RA, Brennan KA, Schenk S, Kennedy MA, Fitzmaurice 

PS. Benzylpiperazine: A drug of abuse? JOURNAL OF 

PSYCHOPHARMACOLOGY 2007;21(8):888-894. 

427. Lanaro R, Costa JL, Zanolli LA, Cazenave SOS. Chemical identification of 

chlorophenylpiperazine in seized tablets. QUIMICA NOVA 

2010;33(3):725-729. 

428. Lecompte Y, Roussel O, Perrin M. 1-Benzylpiperazine (BZP) and 1-(3trifluoromethylphenyl)piperazine 

(TFMPP): Emergence of two agents 

which lead to misuse. ANNALES PHARMACEUTIQUES FRANCAISES 

2008;66(2):85-91. 

429. Maher HM, Awad T, Clark CR. Differentiation of the regioisomeric 2-, 3-, 

and 4-trifluoromethylphenylpiperazines (TFMPP) by GC-IRD and GC-MS. 


430. Nikolova I, Danchev N. Piperazine based substances of abuse: A new 

party pill on Bulgarian drug market. BIOTECHNOLOGY & 

BIOTECHNOLOGICAL EQUIPMENT 2008;22(2):652-655. 

431. Schafstall HJ, Burris MD, Kramer KL, Weston RG. Analytical data for 

ortho-, meta-,and para-chlorophenyl-piperazines (CPP). 2008 American 

Academy of Forensic Sciences Annual Meeting. 

432. Sheridan J, Butler R. "They're legal so they're safe, right?" What did the 

legal status of BZP-party pills mean to young people in New Zealand? 

INTERNATIONAL JOURNAL OF DRUG POLICY 2010;21(1):77-81. 

433. Staack RF. Piperazine designer drugs of abuse. LANCET 

2007;369(9571):1411-1413. 

434. Westphal F, Junge T, Girreser U, Stobbe S, Perez SB. Structure 

elucidation of a new designer benzylpiperazine: 4-Bromo-2,5dimethoxybenzylpiperazine. 

FORENSIC SCIENCE INTERNATIONAL 

2009;187(1-3):87-96. 

435. Wilkins C, Sweetsur P. Differences in harm from legal BZP/TFMPP party 

pills between North Island and South Island users in New Zealand: A case 

of effective industry self-regulation? INTERNATIONAL JOURNAL OF 

DRUG POLICY 2010;21(1):86-90. 

436. Wilkins C, Sweetsur P, Girling M. Patterns of 

benzylpiperazine/trifluoromethylphenylpiperazine party pill use and 

adverse effects in a population sample in New Zealand. DRUG AND 

ALCOHOL REVIEW 2008;27(6):633-639. 

775

1.31 Polydrug: 

437. Laussmann T, Meier-Giebing S. Forensic analysis of hallucinogenic 

mushrooms and khat (Catha edulis Forsk) using cation-exchange liquid 

chromatography. FORENSIC SCIENCE INTERNATIONAL 2010;195(1- 

3):160-164. 

438. Mori T, Ito S, Sawaguchi T. Survey on transactions of methamphetamine 

MDMA and methylphenidate in Japan: A study using the internet. 

INTERNATIONAL MEDICAL JOURNAL 2008;15(1):75-78. 

1.32 Psilocybin Mushrooms, Psilocybin, and Psilocin: 

439. Adamczyk A, Sadakierska-Chudy A, Janoszka J, Rymkiewicz A, Dobosz 

T. Hallucinogenic fungi (psilocybe). Part II. Identification of Psilocybe 

semilanceata by PCR. ARCH MED SADOWEJ KRYMINOLOGIA 

2007;57(3):285. 

440. Boissevain I. Psilocybin mushrooms. TIJDSCHRIFT VOOR 

DIERGENEESKUNDE 2008;133(8):343-343. 

441. Guzman G. Hallucinogenic mushrooms in Mexico: An overview. 

ECONOMIC BOTANY 2008;62(3):404-412. 

442. Guzman G, Gaines RV, Ramirez-Guillen F. New species of hallucinogenic 

Psilocybe (Fr.) P. Kumm. (Agaricomycetideae) from the eastern USA. 

INTERNATIONAL JOURNAL OF MEDICINAL MUSHROOMS 

2007;9(1):75-77. 

443. Guzman G, Kroeger P, Ramirez-Guillen F, Castillo-Del-Moral R. Psilocybe 

(Basidiomycotina, Agaricales, Strophariaceae) in Canada, with a special 

review of species from British Columbia. MYCOTAXON 2008;106:179- 

193. 

444. Horak E, Guzman G, Desjardin DE. Four new species of Psilocybe from 

Malaysia and Thailand, with a key to the species of sect. Neocaledonicae 

and discussion on the distribution of the tropical and temperate species. 

SYDOWIA 2009;61(1):25-37. 

445. Jiang G, Stenzel JR. Identification of psychotropic substances in 

mushrooms by UHPLC/MS. LCGC NORTH AMERICA 2009;(Suppl.)13. 

446. Kocak A, De Cotiis LM, Hoffman DB. Comparative study of ATR and 

transflection IR spectroscopic techniques for the analysis of hallucinogenic 

mushrooms. FORENSIC SCIENCE INTERNATIONAL 2010;195(1-3):36- 

41. 

447. Lott JP, Marlowe DB, Forman RF. Availability of websites offering to sell 

psilocybin spores and psilocybin. JOURNAL OF PSYCHOACTIVE 

DRUGS 2009;41(3):305-307. 

448. Rafati H, Riahi H, Mohammadi A. Enhancement of indole alkaloids 

produced by Psilocybe cubensis (Earle) Singer (Agaricomycetideae) in 

controlled harvesting light conditions. INTERNATIONAL JOURNAL OF 

MEDICINAL MUSHROOMS 2009;11(4):419-426. 

776

449. Stenzel JR, Jiang G. Identification of psychotropic substances in 

mushrooms by UHPLC/MS. LCGC NORTH AMERICA 2009;13. 

1.33 Salvia Divinorum: 

450. Albertson DN, Grubbs LE. Subjective Effects of salvia divinorum: LSD- or 

marijuana-like? JOURNAL OF PSYCHOACTIVE DRUGS 2009;41(3):213- 

217. 

451. Carvalho P, Bikbulatov R, Zjawiony JK, Avery MA. Unusual hemiacetal 

structure derived from Salvinorin A. ACTA CRYSTALLOGRAPHICA 

SECTION E-STRUCTURE REPORTS ONLINE 2008;64:O1370-U2878. 

452. Griffin OH, Miller BL, Khey DN. Legally high? Legal considerations of 

Salvia divinorum. JOURNAL OF PSYCHOACTIVE DRUGS 

2008;40(2):183-191. 

453. Grundmann O, Phipps SM, Zadezensky I, Butterweck V. Salvia divinorum 

and salvinorin A: An update on pharmacology and analytical methodology. 

PLANTA MEDICA 2007;73(10):1039-1046. 

454. Holden KG, Tidgewell K, Marquam A, Rothman RB, Navarro H, Prisinzano 

TE. Synthetic studies of neoclerodane diterpenes from Salvia 

divinorum: Exploration of the 1-position. BIOORGANIC & MEDICINAL 

CHEMISTRY LETTERS 2007;17:6111-6115. 

455. Jermain JD, Evans HK. Analyzing salvia divinorum and its active 

ingredient salvinorin A utilizing thin layer chromatography and gas 

chromatography/mass spectrometry. JOURNAL OF FORENSIC 

SCIENCES 2009;54(3):612-616. 

456. Kennedy JH, Wiseman JM. Direct analysis of Salvia divinorum leaves for 

salvinorin A by thin layer chromatography and desorption electrospray 

ionization multi-stage tandem mass spectrometry. RAPID 

COMMUNICATIONS IN MASS SPECTROMETRY 2010;24(9):1305-1311. 

457. Kutrzeba LM, Ferreira D, Zjawiony JK. Salvinorins J from Salvia 

divinorum: Mutarotation in the neoclerodane system. JOURNAL OF 

NATURAL PRODUCTS 2009;72(7):1361-1363. 

458. Maruyama T, Kamakura H, Kikura-Hanajiri R, Goda Y. Authentication and 

ultra performance liquid chromatography (UPLC)/MS analysis of magic 

mint, Salvia divinorum and its related plants. YAKUGAKU ZASSHI- 


2008;128(1):179-183. 

459. Shirota O, Nagamatsu K, Sekita S. Simple preparative isolation of 

salvinorin a from the hallucinogenic sage, Salvia divinorum, by centrifugal 

partition chromatography. JOURNAL OF LIQUID CHROMATOGRAPHY & 

RELATED TECHNOLOGIES 2007;30(5-8):1105-1114. 

777

460. Simpson DS, Lovell KM, Lozama A, Han N, Day VW, Dersch 

CM, Rothman RB, Prisinzano TE. Synthetic studies of neoclerodane 

diterpenes from Salvia divinorum: Role of the furan in affinity for opioid 

receptors. ORGANIC & BIOMOLECULAR CHEMISTRY 2009;7(18):3748- 

3756. 

461. Tsujikawa K, Kuwayama K, Miyaguchi H, Kanamori T, Iwata YT, Yoshida 

T, Inoue H. Determination of salvinorin A and salvinorin B in Salvia 

divinorum-related products circulated in Japan. FORENSIC SCIENCE 


462. Zhang Y, Chen H, Cheng W, Feng Y, Tang Z, Yang L. Studies on the 

chemical constituents of Salvia miltiorrhiza of Lijiang. ZHONGYAOCAI 

2008;31(2):226-229. 

1.34 Scopolamine: 

463. Henderson TJ, Cullinan DB, Lawrence RJ, Oyler JM. Positive identification 

of the principal component of a white powder as scopolamine by 

quantitative one-dimensional and two-dimensional NMR techniques. 


464. Yuan BQ, Zheng CY, Teng H, You TY. Simultaneous determination of 

atropine, anisodamine, and scopolamine in plant extract by nonaqueous 

capillary electrophoresis coupled with electrochemiluminescence and 

electrochemistry dual detection. JOURNAL OF CHROMATOGRAPHY A 

2010;1217(1):171-174. 

1.35 Steroids: 

465. Blokland MH, Zoontjes PW, Sterk SS, Stephany RW, Zweigenbaum J, van 

Ginkel LA. Confirmatory analysis of trenbolone using accurate mass 

measurement with LC/TOF-MS. ANALYTICA CHIMICA ACTA 

2008;618(1):86-93. 

466. Fragkaki AG, Angelis YS, Tsantili-Kakoulidou A, Koupparis M, 

Georgakopoulos C. Statistical analysis of fragmentation patterns of 

electron ionization mass spectra of enolized-trimethylsilylated anabolic 

androgenic steroids. INTERNATIONAL JOURNAL OF MASS 

SPECTROMETRY 2009;285(1-2):58-69. 

467. Galesio M, Rial-Otero R, Capelo-Martinez JL. Comparative study of 

matrices for their use in the rapid screening of anabolic steroids by matrixassisted 

laser desorption/ionization time-of-flight mass spectrometry. 

RAPID COMMUNICATIONS IN MASS SPECTROMETRY 

2009;23(12):1783-1791. 

468. Hadef Y, Kaloustian J, Portugal H, Nicolay A. Multivariate optimization of a 

derivatisation procedure for the simultaneous determination of nine 

anabolic steroids by gas chromatography coupled with mass 

spectrometry. JOURNAL OF CHROMATOGRAPHY A 2008;1190(1- 

2):278-285. 

778

469. Kauppila TJ, Talaty N, Jackson AU, Kotiaho T, Kostiainen R, Cooks RG. 

Carbohydrate and steroid analysis by desorption electrospray ionization 

mass spectrometry. CHEMICAL COMMUNICATIONS 2008;(23):2674- 

2676. 

470. Morris JA, Minor RG. Confirmation of anabolic steroids by alkaline 

hydrolysis. JOURNAL OF THE CLANDESTINE LABORATORY 


471. Muniz-Valencia R, Gonzalo-Lumbreras R, Santos-Montes A, Izquierdo- 

Hornillos R. Liquid chromatographic method development for anabolic 

steroids using a monolithic column. ANALYTICA CHIMICA ACTA 

2008;611(1):103-112. 

472. Nie J, Wu H, Wang X, Zhang Y, Zhu S, Yu R. Determination of 

testosterone propionate in cosmetics using excitation-emission matrix 

fluorescence based on oxidation derivatization with the aid of secondorder 

calibration methods. ANALYTICA CHIMICA ACTA 2008;628(1):24- 

32. 

473. Parr MK, Zapp J, Becker M, Opfermann G, Bartz U, Schaenzer W. 

Steroidal isomers with uniform mass spectra of their per-TMS derivatives: 

Synthesis of 17-hydroxyandrostan-3-ones, and rost-1-, and -4-ene-3,17diols. 

STEROIDS 2007;72(6-7):545-551. 

474. Pozo OJ, Van Eenoo P, Deventer K, Delbeke FT. Detection and 

characterization of anabolic steroids in doping analysis by LC-MS. TrAC, 

TRENDS IN ANALYTICAL CHEMISTRY 2008;27(8):657-671. 

475. Pozo OJ, Van Eenoo P, Deventer K, Delbeke FT. Ionization of anabolic 

steroids by adduct formation in liquid chromatography electrospray mass 

spectrometry. JOURNAL OF MASS SPECTROMETRY 2007;42(4):497- 

516. 

476. Saudan C, Emery C, Marclay F, Strahm E, Mangin P, Saugy M. Validation 

and performance comparison of two carbon isotope ratio methods to 

control the misuse of androgens in humans. JOURNAL OF 

CHROMATOGRAPHY, B: ANALYTICAL TECHNOLOGIES IN THE 

BIOMEDICAL AND LIFE SCIENCES 2009;877(23):2321-2329. 

477. Shi Y-Q, Yao J, Liu F, Hu C-Q, Yuan J, Zhang Q-M, Jin S-H. 

Establishment of an HPLC identification system for detection of counterfeit 

steroidal drugs. JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL 

ANALYSIS 2008;46(4):663-669. 

478. Silva AI, Pereira HMG, Casilli A, Conceicao FC, Neto FRA. Analytical 

challenges in doping control: Comprehensive two-dimensional gas 

chromatography with time of flight mass spectrometry, a promising option. 

JOURNAL OF CHROMATOGRAPHY A 2009;1216(14):2913-2922. 

479. Stepan R, Cuhra P, Barsova S. Comprehensive two-dimensional gas 

chromatography with time-of-flight mass spectrometric detection for the 

determination of anabolic steroids and related compounds in nutritional 

supplements. FOOD ADDITIVES & CONTAMINANTS, PART A: 

CHEMISTRY, ANALYSIS, CONTROL, EXPOSURE & RISK 

ASSESSMENT 2008;25(5):557-565. 

779

480. Verheyden K, Le Bizec B, Courtheyn D, Mortier V, Vandewiele M, Gillis W, 

Vanthemsche P, De Brabander HF, Noppe H. Mass spectrometric 

detection of and similarities between 1-androgens. ANALYTICA CHIMICA 

ACTA 2007;586(1-2):57. 

481. Walker CJ, Cowan DA, James VH, Lau JC, Kichman AT. Doping in sport- 

2. Quantification of the impurity 19-norandrostenedione in pharmaceutical 

preparations of norethisterone. STEROIDS 2009;74(3):335-340. 

482. Yuan B. Feature analysis and future perspective of designer steroid. 

TIANJIN TIYU XUEYUAN XUEBAO 2007;22(5):422-425. 

483. Yuan Y, Xu C, Peng C, Jin Z, Chen W, Liu L. Analytical methods for the 

detection of corticosteroids-residues in animal-derived foodstuffs. 

CRITICAL REVIEWS IN ANALYTICAL CHEMISTRY 2008;38(4):227-241. 

484. Zhang Y, Tobia HJ, Brenna JT. Steroid isotopic standards for gas 

chromatography-combustion isotope ratio mass spectrometry (GCC- 

IRMS). STEROIDS 2009;74(3):369-378. 

1.36 Synthetic Cannabimimetics (Also “Spice”): 

485. Atwood BK, Huffman J, Straiker A, Mackie K. JWH018, a common 

constituent of 'Spice' herbal blends, is a potent and efficacious 

cannabinoid CB1 receptor agonist. BRITISH JOURNAL OF 

PHARMACOLOGY 2010;160(3):585-593. 

486. Auwaerter V, Dresen S, Weinmann W, Mueller M, Puetz M, Ferreiros N. 

'Spice' and other herbal blends: Harmless incense or cannabinoid 

designer drugs? JOURNAL OF MASS SPECTROMETRY 2009;44(5):832- 

837. 

487. Auwaerter V. Structure elucidation of synthetic materials in “Spice.” 

LABORPRAXIS 2009;33(6):62-64. 

488. Combs M, Morris JA. Analytical profile of two synthetic cannabinoids- 

JWH-018 and JWH-073. JOURNAL OF THE CLANDESTINE 


2008;20(2):2. 

489. Griffiths P, Sedefov R, Gallegos A, Lopez D. How globalization and 

market innovation challenge how we think about and respond to drug use: 

'Spice' a case study. ADDICTION 2010;105(6):951-953. 

490. Hammersley R. Dangers of banning spice and the synthetic cannabinoid 

agonists. ADDICTION 2010;105(2):373-373. 

491. Hermanns-Clausen M, Sauer O, Gerber G, Faerber E, Koch IE, Hentschel 

H, Seidel C, Stedtler U, Auwarter V. New "Herbal Drugs" of Abuse: Spice 

and Smoke. CLINICAL TOXICOLOGY 2009;47(5):57-. 

492. Lindigkeit R, Boehme A, Eiserloh I, Luebbecke M, Wiggermann M, Ernst 

L, Beuerle T. Spice: A never ending story? FORENSIC SCIENCE 


493. Mustata C, Torrens M, Pardo R, Perez C, Farre M. Spice drugs: 

Cannabinoids as new designer drugs. ADICCIONES 2009;21(3):181-186. 

780

494. Piggee C. Investigating a not-so-natural high: Researchers identify 

synthetic cannabinoids in herbal incense. ANALYTICAL CHEMISTRY 

2009;81(9):3205-3207. 

495. Uchiyama N, Kikura-Hanajiri R, Kawahara N, Goda Y. Identification of a 

cannabimimetic indole as a designer drug in a herbal product. FORENSIC 

TOXICOLOGY 2009;27(2):61-66. 

496. Uchiyama N, Kikura-Hanajiri R, Kawahara N, Haishima Y, Goda Y. 

Identification of a cannabinoid analog as a new type of designer drug in a 

herbal product. CHEMICAL & PHARMACEUTICAL BULLETIN 

2009;57(4):439-441. 

497. Uchiyama N, Kikura-Hanajiri R, Ogata J, Goda Y. Chemical analysis of 

synthetic cannabinoids as designer drugs in herbal products. FORENSIC 

SCIENCE INTERNATIONAL 2010;198(1-3):31-38. 

498. Zimmermann US, Winkelmann PR, Pilhatsch M, Nees JA, Spanagel R, 

Schulz K. Withdrawal phenomena and dependence syndrome after the 

consumption of "Spice Gold". DEUTSCHES ARZTEBLATT 


1.37 (Designer) Tryptamines (see also Psilocybin): 

499. Brandt SD, Martins CPB, Freeman S, Dempster N, Riby PG, Gartz J, 

Alder JF. Halogenated solvent interactions with N,N-dimethyltryptamine: 

Formation of quaternary ammonium salts and their artificially induced 

rearrangements during analysis. FORENSIC SCIENCE INTERNATIONAL 

2008;178(2-3):162-170. 

500. Brandt SD, Martins CPB, Freeman S, Dempster N, Wainwright M, Riby 

PG, Alder JF. N,N-Dimethyltryptamine and dichloromethane: 

Rearrangement of quaternary ammonium salt product during GC-EI and 

CI-MS-MS analysis. JOURNAL OF PHARMACEUTICAL AND 

BIOMEDICAL ANALYSIS 2008;47(1):207-212. 

501. Buchanan MS, Carroll AR, Pass D, Quinn RJ. NMR spectral assignments 

of a new chlorotryptamine alkaloid and its analogues from Acacia confusa. 

MAGNETIC RESONANCE IN CHEMISTRY 2007;45(4):359-361. 

502. Cason TD. Mass spectra of designer drugs. JOURNAL OF THE 



503. Chen BH, Liu JT, Chen WX, Chen HM, Lin CH. A general approach to the 

screening and confirmation of tryptamines and phenethylamines by mass 

spectral fragmentation. TALANTA 2008;74(4):512-517. 

504. Fasanello JA, Placke AD. The isolation, identification, and quantitation of 

dimethyltryptamine (DMT) in Mimosa hostilis. MICROGRAM JOURNAL 

2007;5(1-4):41-50. 

505. Gambelunghe C, Aroni K, Rossi R, Moretti L, Bacci M. Identification of 

N,N-dimethyltryptamine and beta-carbolines in psychotropic ayahuasca 

beverage. BIOMEDICAL CHROMATOGRAPHY 2008;22(10):1056-1059. 

781

506. Hsiao Y, Liu JT, Lin CH. Simultaneous separation and detection of 18 

phenethylamine/tryptamine derivatives by liquid chromatography-UV 

absorption and -electrospray ionization mass spectrometry. ANALYTICAL 

SCIENCES 2009;25(6):759-763. 

507. Kamata T, Katagi M, Tsuchihashi H. Metabolism and toxicological 

analyses of hallucinogenic tryptamine analogues being abused in Japan. 

FORENSIC TOXICOLOGY 2010;28(1):1-8. 

508. Martins CPB, Awan MA, Freeman S, Herraiz T, Alder JF, Brandt SD. 

Fingerprint analysis of thermolytic decarboxylation of tryptophan to 

tryptamine catalyzed by natural oils. JOURNAL OF CHROMATOGRAPHY 

A 2008;1210(1):115-120. 

509. Martins CPB, Freeman S, Alder JF, Passie T, Brandt SD. Profiling 

psychoactive tryptamine-drug synthesis by focusing on detection using 

mass spectrometry. TrAC-TRENDS IN ANALYTICAL CHEMISTRY 

2010;29(4):285-296. 

510. Martins CPB, Freeman S, Alder JF, Brandt SD. Characterisation of a 

proposed internet synthesis of N,N-dimethyltryptamine using liquid 

chromatography/electrospray ionisation tandem mass spectrometry. 


511. McIlhenny EH, Pipkin KE, Standish LJ, Wechkin HA, Strassman R, Barker 

SA. Direct analysis of psychoactive tryptamine and harmala alkaloids in 

the Amazonian botanical medicine ayahuasca by liquid chromatographyelectrospray 

ionization-tandem mass spectrometry. JOURNAL OF 


512. Nakamoto A, Namera A, Yahata M, Kuramoto T, Nishida M, Yashiki M. A 

systematic toxicological analysis for hallucinogenic tryptamines in seized 

and biological materials. HIROSHIMA DAIGAKU IGAKU ZASSHI 

2007;55(1-3):1-14. 

513. Ouyang YZ, Liang YZ, Li SH, Luo X, Zhang LX, Tang ZH, Xu XN. 

Interpretation of the characteristic fragmentation mechanisms 

through determining the initial ionization site by natural spin density: 

A study on the derivatives of tryptophan and tryptamine. 

INTERNATIONAL JOURNAL OF MASS SPECTROMETRY 2009;286(2- 

3):112-121. 

514. Pichini S, Pujades M, Marchei E, Pellegrini M, Fiz J, Pacifici R, Zuccaro P, 

Farre M, de la Torre R. Liquid chromatography-atmospheric pressure 

ionization electrospray mass spectrometry determination of 

“hallucinogenic designer drugs” in urine of consumers. JOURNAL OF 

PHARMACEUTICAL AND BIOMEDICAL ANALYSIS 2008;47(2):335-342. 

515. Runguphan W, Maresh JJ, O'Connor SE. Silencing of tryptamine 

biosynthesis for production of nonnatural alkaloids in plant culture. 

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE 

UNITED STATES OF AMERICA 2009;106(33):13673-13678. 

782

516. Salum Pires AP, Rodrigues De Oliveira CD, Moura S, Doerr FA, Silva 

WAE, Yonamine M. Gas chromatographic analysis of dimethyltryptamine 

and beta-carboline alkaloids in ayahuasca, an amazonian psychoactive 

plant beverage. PHYTOCHEMICAL ANALYSIS 2009;20(2):149-153. 

517. Takahashi M, Nagashima M, Suzuki J, Seto T, Yasuda I, Yoshida T. 

Creation and application of psychoactive designer drugs data library using 

liquid chromatography with photodiode array spectrophotometry detector 

and gas chromatography-mass spectrometry. TALANTA 2009;77(4):1245- 

1272. 

518. Takahashi M, Nagashima M, Suzuki J, Seto T, Yasuda I, Yoshida T. 

Analysis of phenethylamines and tryptamines in designer drugs using gas 

chromatography-mass spectrometry. JOURNAL OF HEALTH SCIENCE 

2008;54(1):89-96. 

519. Uchiyama N, Kikura-Hanajiri R, Kawahara N, Goda Y. Analysis of 

designer drugs detected in the products purchased in fiscal year 2006. 


OF JAPAN 2008;128(10):1499-1505. 

520. Uchiyama N, Miyazawa N, Kawamura M, Kikura-Hanajiri R, Goda Y. 

Analysis of newly distributed designer drugs detected in the products 

purchased in fiscal year 2008. YAKUGAKU ZASSHI-JOURNAL OF THE 

PHARMACEUTICAL SOCIETY OF JAPAN 2010;130(2):263-270. 

521. Wang M-J, Liu J-T, Chen H-M, Lin J-J, Lin C-H. Comparison of the 

separation of nine tryptamine standards based on gas chromatography, 

high performance liquid chromatography and capillary electrophoresis 

methods. JOURNAL OF CHROMATOGRAPHY A 2008;1181(1-2):131- 

136. 

522. Wang MJ, Tsai CH, Hsu WY, Liu JT, Lin CH. Optimization of separation 

and online sample concentration of N,N-dimethyltryptamine and related 

compounds using MEKC. JOURNAL OF SEPARATION SCIENCE 

2009;32(3):441-445. 

523. Westphal F, Junge T, Roesner P, Fritschi G, Klein B, Girreser U. Mass 

spectral and NMR spectral data of two new designer drugs with an αaminophenone 

structure: 4'-Methyl-α-pyrrolidinohexanophenone and 4'methyl-α-pyrrolidinobutyrophenone. 

FORENSIC SCIENCE 

INTERNATIONAL 2007;169(1):32. 

524. Wolf EU, Raziel A, Katz Y. The “periodic table” of designer drugs in Israel. 

JOURNAL OF THE CLANDESTINE LABORATORY INVESTIGATING 

CHEMISTS ASSOCIATION 2008;20(2):19-20. 

525. Zaitsu K, Katagi M, Kamata T, Kamata H, Shima N, Tsuchihashi 

H, Hayashi T, Kuroki H, Matoba R. Determination of a newly encountered 

designer drug "p-methoxyethylamphetamine" and its metabolites in human 

urine and blood. FORENSIC SCIENCE INTERNATIONAL 

2008;177(1):77-84. 

783

1.38 Zaleplon: 

526. Bharathi C, Prabahar KJ, Prasad CS, Kumar MS, Magesh S, Handa VK, 

Dandala R, Naidu A. Impurity profile study of Zaleplon. JOURNAL OF 

PHARMACEUTICAL AND BIOMEDICAL ANALYSIS 2007;44(1):101-109. 

527. Metwally FH, Abdelkawy M, Abdelwahab NS. Application of 

spectrophotometric, densitometric, and HPLC techniques as stability 

indicating methods for determination of Zaleplon in pharmaceutical 

preparations. SPECTROCHIMICA ACTA PART A-MOLECULAR AND 

BIOMOLECULAR SPECTROSCOPY 2007;68(5):1220-1230. 

528. Radl S, Blahovcova M, Placek L, Pekarek T, Havlicek J. Synthesis of 

some impurities and/or degradation products of Zaleplon. JOURNAL OF 

HETEROCYCLIC CHEMISTRY 2010;47(2):276-283. 

1.39 Zolpidem: 

529. Emshanova SV, Veselova NI, Zuev AP, Sadchikova NP. Direct molding 

technology for the production of Zolpidem tablets. KHIMIKO- 

FARMATSEVTICHESKII ZHURNAL 2007;41(12):37-39. 

530. Halasz I, Dinnebier RE. Structural and thermal characterization of 

Zolpidem hemitartrate hemihydrate (form e) and its decomposition 

products by laboratory x-ray powder diffraction. JOURNAL OF 

PHARMACEUTICAL SCIENCES 2010;99(2):871-878. 

531. Owen RT. Novel Zolpidem formulations. DRUGS OF TODAY 

2009;45(5):395-400. 

532. Pushpalatha P, Sarin RK, Rao MA, Baggi TRR. A new thin-layer 

chromatographic method for analysis of Zolpidem and Zopiclone. JPC- 

JOURNAL OF PLANAR CHROMATOGRAPHY-MODERN TLC 

2009;22(6):449-451. 

533. Sumalatha Y, Reddy PP, Reddy R, Satyanarayana B. Synthesis and 

spectral characterization of process-related substances to the hypnotic 

agent Zolpidem. ARKIVOC 2009;143-149. 

534. Sumalatha Y, Reddy TR, Reddy PP, Satyanarayana B. A simple, efficient 

and scalable synthesis of hypnotic agent, Zolpidem. ARKIVOC 2009;315- 

320. 

1.40 Zopiclone: 

535. Gomis DB, Velasco CB, Sanchez IH, Alvarez MDG. Optimization by 

factorial design of a capillary electrophoresis method for the chiral 

resolution and determination of Zopiclone and its synthesis precursor. 

JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED 

TECHNOLOGIES 2009;32(18):2654-2668. 

784

536. Chen YY, Liu JJ, Tang KW. Study on inclusion complexes of betacyclodextrin 

with Zopiclone. SPECTROSCOPY AND SPECTRAL 

ANALYSIS 2008;28(10):2380-2383. 

537. Jerussi TP, Fang QK. Compositions comprising Zopiclone derivatives and 

methods of making and using the same. PN US 07456173 AE Sepracor 

Inc. Official Gazette of the United States Patent and Trademark Office 

Patents 2008. 

538. Sangaraju S, Kanth ML, Rao BM, Someswararao N. Enantiomeric 

separation of S-Zopiclone and its R-enantiomer in bulk drug samples by 

validated chiral RP-HPLC. PHARMAZIE 2009;64(11):717-719. 

539. Yilmaz S. Adsorptive stripping voltammetric determination of Zopiclone in 

tablet dosage forms and human urine. COLLOIDS AND SURFACES B: 

BIOINTERFACES 2009;71(1):79-83. 

1.41 Miscellaneous: 

540. Suzuki J, Takahashi M, Nagashima M, Seto T, Mori K, Ogino S. 

Determination of chloral hydrate in unapproved/unpermitted drugs. 

TOKYO-TO KENKO ANZEN KENKYU SENTA KENKYU NENPO 

2009;59:79-84. 

541. Tcheremissine OV. Is quetiapine a drug of abuse? Reexamining the issue 

of addiction. EXPERT OPINION ON DRUG SAFETY 2008;7(6):739-48. 

2. Synthesis and/or Cultivation of Abused Substances, their 

Issue: 

Precursors, and Essential Chemicals 

Forensic chemists must maintain familiarity with existing and new clandestine 

syntheses of abused substances, their precursors, and essential chemicals, and 

with the cultivation of abused natural products, in order to assist enforcement 

activities, to ensure safety and effectiveness during enforcement operations, and 

to provide expert testimony in legal proceedings. 

Solution: 

Illicit drug seizures, clandestine laboratory operations, and illicit grow operations, 

are continuously monitored to maintain a comprehensive overview of the field. In 

cases where new drugs are synthesized, or new methodologies are utilized, case 

reports are generated for the forensic and enforcement communities. 

785

2.1 Production of Abused Substances and/or their Precursors and 

Essential Chemicals: 

542. Alacid E, Najera C. Regioselective Heck reaction of N-vinylphthalimide: A 

general strategy for the synthesis of (E)-N-styrylphthalimides and 

phenethylamines. ADVANCED SYNTHESIS & CATALYSIS 

2008;350(9):1316-1322. 

543. Allwein SP, McWilliams JC, Secord EA, Mowrey DR, Nelson TD, Kress 

MH. Efficient synthesis of chiral phenethylamines: Preparation, 

asymmetric hydrogenation, and mild deprotection of enetrifluoroacetamides. 

TETRAHEDRON LETTERS 2006;47:6409-6412. 

544. Barreto RD, Nascimbem LBLR, Correia CRD. Synthesis of the 3-(3,4,5trimethoxyphenyl)pyrrolidine: 

A new conformationally constrained 

mescaline analogue. SYNTHETIC COMMUNICATIONS 2007;37:2011- 

2018. 

545. Bergman YE, Mulder R, Perlmutter P. Total synthesis of 20-Norsalvinorin 

A. 1. Preparation of a key intermediate. JOURNAL OF ORGANIC 

CHEMISTRY 2009;74(6):2589-2591. 

546. Bikbulatov RV, Stewart J, Jin WT, Yan F, Roth BL, Ferreira D, Zjawiony 

JK. Short synthesis of a novel class of salvinorin A analogs 

with hemiacetalic structure. TETRAHEDRON LETTERS 2008;49(6):937- 

940. 

547. Brandt SD, Tirunarayanapuram SS, Freeman S, Dempster N, Barker SA, 

Daley PF, Cozzi NV, Martins CPB. Microwave-accelerated synthesis of 

psychoactive deuterated N,N-dialkylated-[alpha,alpha,beta,beta-d(4)]tryptamines. 

JOURNAL OF LABELLED COMPOUNDS AND 

RADIOPHARMACEUTICALS 2008;51(13-14):423-429. 

548. Carroll RJ, Leisch H, Rochon L, Hudlicky T, Cox DP. One-pot conversion 

of thebaine to hydrocodone and synthesis of neopinone ketal. JOURNAL 

OF ORGANIC CHEMISTRY 2009;74(4):1812-1812. 

549. Chen YW, Gao MZ, Tan SD, Reibenspies JH, Zingaro RA. Synthesis and 

structures of new C-2-symmetrical chiral macrocycles containing the 

ephedrine moiety. HETEROCYCLES 2009;78(4):891-897. 

550. Cloonan SM, Keating JJ, Corrigan D, O'Brien JE, Kavanagh PV, Williams 

DC, Meegan MJ. Synthesis and in vitro toxicity of 4-MTA, its characteristic 

clandestine synthesis byproducts and related sulfur substituted alphaalkylthioamphetamines. 

BIOORGANIC & MEDICINAL CHEMISTRY 

2010;18(11):4009-4031. 

551. Coote SJ, Davies SG, Fletcher AM, Roberts PM, Thomson JE. 

Enantiospecific Stereodivergent synthesis of trans- and cis-N(2),3dimethyl-4-phenyl-1,2,3,4-tetrahydroisoquinolines. 

CHEMISTRY-AN 

ASIAN JOURNAL 2010;5(3):589-604. 

552. Cox M. The source of tolualdehyde derived oxazolidines in clandestine 

laboratory samples. AUSTRALIAN JOURNAL OF FORENSIC SCIENCES 

2008;40(2):161-170. 

786

553. Cox M, Klass G, Wei Min Koo C. Forensic aspects of the 

biotransformation of benzaldehyde used in the synthesis of 

methamphetamine, Part 1: Reaction conditions, stereochemical outcomes, 

and the use of other substituted benaldehydes. JOURNAL OF THE 



554. Dal Cason TA. Synthesis and identification of N,N-dimethylcathinone 

hydrochloride. MICROGRAM JOURNAL 2007;5(1-4):3-12. 

555. Davis SF, Culshaw PN, Wermuth UD. The production of phenyl-2propanone 

from benzaldehyde via a baeyer-villiger reaction. JOURNAL 

OF THE CLANDESTINE LABORATORY INVESTIGATING CHEMISTS 


556. Edwankar CR, Edwankar RV, Namjoshi OA, Rallapalli SK, Yang J, Cook 

JM. Recent progress in the total synthesis of indole alkaloids. CURRENT 

OPINION IN DRUG DISCOVERY & DEVELOPMENT 2009;12(6):752- 

771. 

557. Gould C. iodine recovery from povidone-iodine solutions and its use in 

clandestine methamphetamine synthesis. 2008 American Academy of 


558. Groeper JA, Eagles JB, Hitchcock SR. A facile, one-pot synthesis of 

Ephedra-based aziridines. TETRAHEDRON-ASYMMETRY 

2009;20(17):1969-1974. 

559. Guizzetti S, Benaglia M, Biaggi C, Celentano G. A convenient, highly 

stereoselective, metal-free synthesis of chiral amines. SYNLETT 

2010;(1):134-136. 

560. Guy M, Freeman S, Alder JF, Brandt SD. The Henry Reaction: 

Spectroscopic studies of nitrile and hydroxylamine by-products formed 

during synthesis of psychoactive phenylalkylamines. CENTRAL 

EUROPEAN JOURNAL OF CHEMISTRY 2008;6(4):526-534. 

561. Hagiwara H, Nozawa M. Highly oxygenated diterpenoids associated to the 

central nervous system: Syntheses of salvinorin and forskolin. JOURNAL 

OF SYNTHETIC ORGANIC CHEMISTRY JAPAN 2009;67(11):1115-1124. 

562. Hagiwara H, Suka Y, Nojima T, Hoshi T, Suzuki T. Second-generation 

synthesis of salvinorin A. TETRAHEDRON 2009;65(25):4820-4825. 

563. Hu CM, Qin H, Cui YX, Jia YX. Palladium-catalyzed synthesis of 

tryptamines and tryptamine homologues: Synthesis of psilocin. 

TETRAHEDRON 2009;65(45):9075-9080. 

564. Inuki S, Oishi S, Fujii N, Ohno H. Total synthesis of (+/-)-lysergic acid, 

lysergol, and isolysergol by palladium-catalyzed domino cyclization of 

amino allenes bearing a bromoindolyl group. ORGANIC LETTERS 

2008;10(22):5239-5242. 

565. Kesharwani S, Sahu NK, Kohli DV. Synthesis And biological evaluation of 

some new spiro derivatives of barbituric acid. PHARMACEUTICAL 

CHEMISTRY JOURNAL 2009;43(6):315-319. 

787

566. Kok G, Ashton TD, Scammells PJ. An improved process for the Ndemethylation 

of opiate alkaloids using an iron(ii) catalyst in acetate 

buffer. ADVANCED SYNTHESIS & CATALYSIS 2009;351(1-2):283-286. 

567. Kurokawa T, Isomura M, Tokuyama H, Fukuyama T. Synthesis of lysergic 

acid methyl ester via the double cyclization strategy. SYNLETT 

2009;(5):775-778. 

568. Lozama A, Prisinzano TE. Chemical methods for the synthesis and 

modification of neoclerodane diterpenes. BIOORGANIC & MEDICINAL 

CHEMISTRY LETTERS 2009;19(18):5490-5495. 

569. Ma J, Yin WY, Zhou H, Liao XB, Cook JM. General approach to the total 

synthesis of 9-methoxy-substituted indole alkaloids: Synthesis of 

mitragynine, as well as 9-methoxygeissoschizol and 9-methoxy-N-bmethylgeissoschizol. 

JOURNAL OF ORGANIC CHEMISTRY 

2009;74(1):264-273. 

570. Ma ZZ, Lee DYW. Synthesis of deacetyl-1,10-didehydrosalvinorin G. 

TETRAHEDRON LETTERS 2008;49(11):1782-1785. 

571. Magnus P, Sane N, Fauber BP, Lynch V. Concise syntheses of (-)galanthamine 

and (+/)-codeine via intramolecular alkylation of a phenol 

derivative. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 

2009;131(44):16045. 

572. Mayo E, Coxon A, Johnson C. The reduction of pseudoephedrine to 

methamphetamine using phosphorous acid and iodine. JOURNAL OF 



573. Michaelis DJ, Dineen TA. Ring opening of aziridines with orthobromophenyl 

metal reagents: Synthesis of 2-substituted indolines. 

TETRAHEDRON LETTERS 2009;50(17):1920-1923. 

574. Nicolaou KC, Krasovskiy A, Majumder U, Trepanier VE, Chen DYK. New 

synthetic technologies for the construction of heterocycles 

and tryptamines. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 

2009;131(10):3690-3699. 

575. Nicolaou KC, Krasovskiy A, Trepanier VE, Chen DYK. An expedient 

strategy for the synthesis of tryptamines and other heterocycles. 

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION 2008;47(22):4217- 

4220. 

576. Norman K. The synthesis of amphetamine and methamphetamine: A “big” 

picture. JOURNAL OF THE CLANDESTINE LABORATORY 

INVESTIGATING CHEMISTS ASSOCIATION 2008;19(3):10-29. 

577. Nozawa M, Suka Y, Hoshi T, Suzuki T, Hagiwara H. Total synthesis of the 

hallucinogenic neoclerodane diterpenoid salvinorin A. ORGANIC 

LETTERS 2008;10(7):1365-1368. 

578. Painter B, Pigou PE, W.M. Koo C. The Willgerodt-Kindler reaction. Part 1: 

Styrene to methamphetamine. JOURNAL OF THE CLANDESTINE 


2008;19(4):53. 

788

579. Pan ZR, Zheng HG, Wang TW, Song Y, Li YZ, Guo ZJ, Batten SR. 

Hydrothermal synthesis, structures, and physical properties of four new 

flexible multicarboxylate ligands-based compounds. INORGANIC 

CHEMISTRY 2008;47(20):9528-9536. 

580. Pearson JR, Reid EF, Rowe JE. The preparation of y-butyrolactone from 

readily available starting materials. JOURNAL OF THE CLANDESTINE 


2008;19(1):8. 

581. Refat MS, EI-Korashy SA, Ahmed AS. A convenient method for the 

preparation of barbituric and thiobarbituric acid transition metal 

complexes. SPECTROCHIMICA ACTA PART A-MOLECULAR AND 

BIOMOLECULAR SPECTROSCOPY 2008;71(3):1084-1094. 

582. Scheerer JR, Lawrence JF, Wang GC, Evans DA. Asymmetric synthesis 

of salvinorin A, a potent kappa opioid receptor agonist. JOURNAL OF 

THE AMERICAN CHEMICAL SOCIETY 2007;129:8968. 

583. Sengmany S, Le Gall E, Le Jean C, Troupel M, Nedelec JY. 

Straightforward three-component synthesis of diarylmethylpiperazines and 

1,2-diarylethylpiperazines. TETRAHEDRON 2007;63:3672-3681. 

584. Stork G, Yamashita A, Adams J, Schulte GR, Chesworth R, Miyazaki 

Y, Farmer JJ. Regiospecific and stereoselective syntheses of (+/-) 

morphine codeine and thebaine via a highly stereocontrolled 

intramolecular 4+2 cycloaddition leading to a phenanthrofuran system. 

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 

2009;131(32):11402-11406. 

585. Toske SG, Hays PA, Geer BL. The synthesis and identification of Nacetylpseudoephedrine 

and N-acetylephedrine. JOURNAL OF THE 



586. Walker AR, Love DW, Bordelon JA. Phosphorous acid flakes used as a 

substitute for red phosphorus in the reduction of (pseudo) ephedrine to 

methamphetamine. JOURNAL OF THE CLANDESTINE LABORATORY 


587. Yamauchi T, Hagiwara S, Higashiyama K. Concise Total synthesis of (-)deoxocuscohygrine 

and (-)-dihydrocuscohygrine. JOURNAL OF 

ORGANIC CHEMISTRY 2008;73(24):9784-9787. 

588. Yan F, Bikbulatov RV, Mocanu V, Dicheva N, Parker CE, Wetsel 

WC, Mosier PD, Westkaemper RB, Allen JA, Zjawiony JK, Roth BL. 

Structure-based design, synthesis, and biochemical and 

pharmacological characterization of novel salvinorin a analogues as active 

state probes of the kappa-opioid receptor. BIOCHEMISTRY 

2009;48(29):6898-6908. 

589. Yang L, Xu W, Chen F, Liu-Chen LY, Ma ZZ, Lee DYW. Synthesis and 

biological evaluation of C-12 triazole and oxadiazole analogs of salvinorin 

A. BIOORGANIC & MEDICINAL CHEMISTRY LETTERS 

2009;19(5):1301-1304. 

789

590. Yun H, Kim BG. Enzymatic production of (R)-phenylacetylcarbinol by 

pyruvate decarboxylase from Zymomonas mobilis. BIOTECHNOLOGY 

AND BIOPROCESS ENGINEERING 2008;13(3):372-376. 

591. Zhang L, Ding ZY, Shi GY. Asymmetric biosynthesis of (1S,2S)-ephedrine 

by Morganella morganii CMCC(B)49208. AFRICAN JOURNAL OF 

BIOTECHNOLOGY 2009;8(4):694-698. 

3. Clandestine Laboratories - Appraisals and Safety 

Issue: 

Forensic chemists must maintain familiarity with clandestine laboratory 

procedures, setups, and techniques in order to assist enforcement activities, to 

ensure safety and effectiveness during enforcement operations, and in order to 

provide expert testimony in court proceedings. 

Solution: 

Clandestine laboratory operations are continuously reviewed to provide a 

comprehensive overview of the field. In cases where new methodologies are 

noted, or unusual safety concerns are salient, reports are generated for the 

forensic and enforcement communities. 

3.1 General 

592. Coxon A, McLeay N, Ranaweera N. Development of scene examination 

software for clandestine drug laboratory scenes. JOURNAL OF THE 



593. Coxon A. The cost of “p-lab” callouts. JOURNAL OF THE CLANDESTINE 


2008;18(1):10. 

594. MacFarlane KJ, Neely IJ. Identification of hypophosphorous and 

hydrochloric acid mixtures encountered at clandestine laboratories. 


CHEMISTS ASSOCIATION 2008;19(2):16. 

595. Man G, Stoeber B, Walus K. An assessment of sensing technologies for 

the detection of clandestine methamphetamine drug laboratories. 


596. Montefusco V. An improved scent transfer device for collecting evidence 

at a crime scene. U.S. Pat. Appl. Publ. US 2008 87,110. 

790

597. Norman K. Improvised containers used for hydrogen chloride gas 

generation at a clandestine drug laboratory. JOURNAL OF THE 



598. Solomon KR, Anadon A, Brain RA, Cerdeira AL, Crossan AN, Marshall J, 

Sanin LH, Smith L. Comparative hazard assessment of the substances 

used for production and control of coca and poppy in colombia. 

RATIONAL ENVIRONMENTAL MANAGEMENT OF AGROCHEMICALS: 

RISK ASSESSMENT, MONITORING, AND REMEDIAL ACTION 

2007;966:87-99. 

599. Solomon KR, Marshall EJP. Production of illicit drugs, the environment, 

and human health. JOURNAL OF TOXICOLOGY AND 

ENVIRONMENTAL HEALTH-PART A-CURRENT ISSUES 2009;72(15- 

16):913-913. 

600. Wong L, Sannerud C, Carr S. Clandestine laboratory, synthetic drugs and 

precursor chemicals trends in the united states over a seven year period, 

2001-2007. 2008 American Academy of Forensic Sciences Annual 

Meeting. 

3.2 Clandestine Laboratory Appraisals and Safety: 

601. Abdullah AFL, Miskelly GM. Recoveries of trace pseudoephedrine and 

methamphetamine residues from impermeable household surfaces: 

Implications for sampling methods used during remediation of clandestine 

methamphetamine laboratories. TALANTA 2010;81(1-2):455-461. 

602. Cotner J, Cole B, Healy J. Quantitative comparison of common 

decontamination methods. JOURNAL OF THE CLANDESTINE 


2008;18(1):6. 

603. McLeod V, Ketcham G. The safety guys: Introduction to clandestine drug 

laboratories - A serious health and safety concern. FORENSIC 

MAGAZINE 2007;4(3):48. 

604. Patrick G, Daniell W, Treser C. Residual methamphetamine in 

decontaminated clandestine drug laboratories. JOURNAL OF 

OCCUPATIONAL AND ENVIRONMENTAL HYGIENE 2009;6(3):151-156. 

605. VanDyke M, Erb N, Arbuckle S, Martyny J. A 24-hour study to investigate 

persistent chemical exposures associated with clandestine 

methamphetamine laboratories. JOURNAL OF OCCUPATIONAL AND 

ENVIRONMENTAL HYGIENE 2009;6(2):82-89. 

3.3 Safety Issues - Case Reports: 

606. Burge M, Hunsaker JC, Davis GJ. Death of a toddler due to ingestion of 

sulfuric acid at a clandestine home methamphetamine laboratory. 

JOURNAL OF BIOLOGICAL CHEMISTRY 2010;285(2):298-301. 

791

607. Grant P, Bell K, Stewart D, Paulson J, Rogers K. Evidence of 

methamphetamine exposure in children removed from clandestine 

methamphetamine laboratories. PEDIATRIC EMERGENCY CARE 

2010;26(1):10-14. 

608. Savopolos, JA, Person EC. Date rape drugs and children’s toys. 



609. Wells SM, Noonan C, Wells KM, Holian A, Wibbemmeyer LA. Effects of 

toxic gases: methamphetamine inhalation. JOURNAL OF BURN CARE & 

RESEARCH 2009;30(1):152-154. 

610. Witter RZ, Martyny JW, Mueller K, Gottschall B, Newman LS. Symptoms 

experienced by law enforcement personnel during methamphetamine lab 

investigations. JOURNAL OF OCCUPATIONAL AND ENVIRONMENTAL 

HYGIENE 2007;4:895-902. 


611. Frish MB, Wainner RT, Laderer MC, Green BD, Allen MG. Standoff and 

miniature chemical vapor detectors based on tunable diode laser 

absorption spectroscopy. IEEE SENSORS JOURNAL 2010;10(3):639- 

646. 

612. Zipf EC. Emission detector for the remote detection of explosives and 

illegal drugs. (Patent) CHEMICAL ABSTRACTS 2007:874282. 

4 Reference Drug Standards and Total Syntheses 

Issue: 

Many reference drug standards or structurally related internal standards are 

either commercially unavailable, or if available are extremely expensive. 

Solution: 

Controlled substances and their structural or isotopically labelled analogs are 

synthesized as needed. Internal standards are also prepared as needed. Case 

reports are published for new or unusual standards or improved synthetic 

approaches. 

4.1 General 

608. Rundlof T, Mathiasson M, Bekiroglu S, Hakkarainen B, Bowden T, 

Arvidsson T. Survey and qualification of internal standards for 

quantification by H-1 NMR spectroscopy. JOURNAL OF 


792

609. Schwarz M, Klier B, Sievers H. Herbal reference standards. PLANTA 

MEDICA 2009;75(7):689-703. 

610. Shaikh AC, Chen CP. Synthesis of deuterium-labeled Zaleplon-d(5) as an 

internal standard. JOURNAL OF LABELLED COMPOUNDS & 

RADIOPHARMACEUTICALS 2008;51(1-2):72-76. 

611. Shaikh AC, Wang YY, Chen CP. Synthesis of deuterium-labelled 

standards of (+/-)-DOM and (+/-)-MMDA. JOURNAL OF LABELLED 

COMPOUNDS & RADIOPHARMACEUTICALS 2007;50(7-8):660-665. 

612. Sharpless KE, Duewer DL. Standard reference materials for analysis of 

dietary supplements. JOURNAL OF AOAC INTERNATIONAL 

2008;91(6):1298-1302. 

613. Sharpless KE, Thomas JB, Christopher SJ, Greenberg RR, Sander 

LC, Schantz MM, Welch MJ, Wise SA. Standard reference materials for 

foods and dietary supplements. ANALYTICAL & BIOANALYTICAL 

CHEMISTRY 2007;389:171-178. 

614. Wang YY, Chen C. Synthesis of deuterium labeled tryptamine derivatives. 

JOURNAL OF THE CHINESE CHEMICAL SOCIETY 2007;54(5):1363- 

1368. 

615. Wang YY, Chen CP. Synthesis of a deuterium-labelled standard of 

bufotenine (5-HO-DMT). JOURNAL OF LABELLED COMPOUNDS & 

RADIOPHARMACEUTICALS 2007;50(13-14):1262-1265. 

616. You ZY, Chen YJ, Wang YY, Chen CP. Synthesis of deuterium labeled 

standards of 1-benzylpiperazine, fenetylline, nicocodeine and 

nicomorphine. JOURNAL OF THE CHINESE CHEMICAL SOCIETY 

2008;55(3):663-667. 

5. Source Determination of Drugs (Impurity Profiling) and Comparative 

Issues: 

Analyses 

Impurity profiling of drugs is important for comparative analysis protocols, geosourcing, 

and synthetic route determinations. However, although certain drugs 

have been well characterized with respect to their impurity profiles, most have 

not been properly investigated. 

Comparative analysis (i.e., the systematic application of impurity profiling for 

determination of commonality of origin) is complicated due to both the high 

complexity of the data and the large numbers of exhibits. Improved analytical 

and data handling techniques are needed. 

793

Solution: 

High sensitivity analytical techniques (primarily chromatographic) provide 

detailed profiles of trace-level impurities, ions, trace metals, and stable isotopes. 

Identification of individual impurities enhance origin identification and 

comparative analyses and also aid in development of internal standards for 

improved accuracy and precision of analysis. 

In-depth analysis via improved instrumental methodologies help identify 

discriminatory components in impurity profiles. Computer databases, sorting 

programs, and pattern recognition/neural networks provide enhanced data 

handling and analysis, enabling and improving comparative analyses. 

Case reports are generated for the forensic and enforcement communities. 

5.1 Amphetamine(s): 

617. Kokiashvili NG, Wongwan S, Landgraf C, Michaelis K, Hammitzsch- 

Wiedemann M, Scriba GKE. Profiling of levoamphetamine and related 

substances in dexamphetamine sulfate by capillary electrophoresis. 

JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS 

2009;50(5):1050-1053. 

618. Zhang J, Zhang D. Study on the optimization methods of common-batch 

identification of amphetamine samples. WEISHENG YANJIU 

2008;37(4):505-507. 

5.2 Cocaine: 

619. Dujourdy L, Besacier F. Headspace profiling of cocaine samples for 

intelligence purposes. FORENSIC SCIENCE INTERNATIONAL 

2008;179(2-3):111-122. 

620. Galimov EM, Sevast’yanov VS, Kul’bachevskaya EV, Golyavin AA. The 

identification of the geographical origin of narcotic drugs based on the 

carbon and nitrogen isotope ratios. MASS-SPEKTROMETRIYA 

2004;1(1):31. 

621. Garzon WF, Parada F, Florian NM. Forensic analysis of cocaine samples 

produced in colombia: I. Chromatographic profiling. VITAE-REVISTA DE 

LA FACULTAD DE QUIMICA FARMACEUTICA 2009;16(2):228-236. 

622. Garzon WF, Parada F, Florian NM. Forensic analysis of cocaine samples 

produced in colombia: I. Chromatographic profiling. VITAE-REVISTA DE 

LA FACULTAD DE QUIMICA FARMACEUTICA 2009;16(2):220-227. 

623. Lociciro S, Hayoz P, Esseiva P, Dujourdy L, Besacier F, Margot P. 

Cocaine profiling for strategic intelligence purposes, a cross-border project 

between France and Switzerland - Part I. Optimisation and harmonisation 

of the profiling method. FORENSIC SCIENCE INTERNATIONAL 

2007;167(2-3):220-228. 

794

624. Lociciro S, Esseiva P, Hayoz P, Dujourdy L, Besacier F, Margot P. 

Cocaine profiling for strategic intelligence, a cross-border project between 

France and Switzerland - Part II. Validation of the statistical methodology 

for the profiling of cocaine. FORENSIC SCIENCE INTERNATIONAL 

2008;177(2-3):199-206. 

625. Sewenig S, Fichtner S, Holdermann T, Fritschi G, Neumann H. 

Determination of delta C-13(V-PDB) and delta N-15(AIR) values 

of cocaine from a big seizure in Germany by stable isotope ratio 

mass spectrometry. ISOTOPES IN ENVIRONMENTAL AND HEALTH 

STUDIES 2007;43(4):275-280. 

5.3 Heroin: 

626. Cai XL, Wu GP. Preliminary study on identification of heroin from different 

routes with clustering analysis by Fourier transform infrared spectroscopy. 

SPECTROSCOPY AND SPECTRAL ANALYSIS 2007;27(12):2441-2444. 

627. Dufey V, Dujourdy L, Besacier F, Chaudron H. A quick and automated 

method for profiling heroin samples for tactical intelligence purposes. 


628. Hibbert DB, Blackmore D, Li JF, Ebrahimi D, Collins M, Vujic S, 

Gavoyannis P. A probabilistic approach to heroin signatures. 

ANALYTICAL & BIOANALYTICAL CHEMISTRY 2010;396(2):765-773. 

629. Huai C, Chen G, Hong L, Fan L, Ping F, Zhao W, Yang H, Su S, Wen Y. A 

new method to differentiate heroin source and the use of software. 

JISUANJI YU YINGYONG HUAXUE 2008;25(11):1329-1333. 

630. Lurie IS, Toske SG. Applicability of ultra-performance liquid 

chromatography-tandem mass spectrometry for heroin profiling. 


631. Morello DR, Cooper SD, Panicker S, Casale JF. Signature profiling and 

classification of illicit heroin by gc-ms analysis of acidic and neutral 

manufacturing impurities. JOURNAL OF FORENSIC SCIENCES 

2010;55(1):42-49. 

632. Nguyen XT, Hoang MH, Do DN, Tran VS. Establishment of the method for 

analysis of solvent residue in heroin samples to track the origin. TAP CHI 

DUOC HOC 2007;47(2):34. 

633. Odell LR, Skopec J, McCluskey A. Isolation and identification of unique 

marker compounds from the Tasmanian poppy Papaver somniferum N. 

Implications for the identification of illicit heroin of Tasmanian origin. 


634. Odell LR, Skopec J, McCluskey A. Isolation and identification of unique 

marker compounds from the Tasmanian poppy Papaver somniferum N. 

Implications for the identification of illicit heroin of Tasmanian origin. 


635. Qian Z, Zhang C, Zheng H. Determination on heroin geographical source 

by capillary electrophoresis and principal component analysis. HUAXUE 

FENXI JILIANG 2008;17(2):31-33. 

795

636. Qian Z, Zhang C, Zheng H. Determination of heroin geographical source 

by capillary electrophoresis and discriminant analysis. HUAXUE FENXI 

JILIANG 2008;17(4):40-42. 

637. Wang Y, Wen Y, Luo A, Zou H. Application of hierarchical clustering to the 

classification of spectra of pyrolytic cracking of heroin. LIHUA JIANYAN, 

HUAXUE FENCE 2008;44(3):205-208. 

638. Wen Y, Zhang J. Testing of 15 elements in heroin produced in different 

areas by ICP-AES. LIHUA JIANYAN, HUAZUE FENCE 2008;44(10):980- 

985. 

639. Xing H, Ma G, Zhang C, Bai Y. Application of chemometrics in heroin 

source determination. JISUANJI YU YINGYONG HUAXUE 

2008;25(4):437-440. 

640. Zhang C, Zheng H, Qian Z. Rapid test and source determination of heroin 

samples by GC/FID. HUAXUE FENXI JILIANG 2007;16(5):39-41. 

5.4 Marijuana: 

641. Ehleringer JR, West J, Hurley J. Geographical region of origin and 

cultivation assignments of marijuana seizures in major cities. The Fourth 

Forensic Isotope Ratio Mass Spectrometry Conference: 2010. 

642. ElSohly MA, Stanford DF, Murphy TP. Chemical fingerprinting of cannabis 

as a means of source identification. FORENSIC SCIENCE AND 

MEDICINE 2007;51-66. 

643. Gilmore S, Peakall R, Robertson J. Organelle DNA haplotypes reflect 

crop-use characteristics and geographic origins of Cannabis sativa. 

FORENSIC SCIENCE INTERNATIONAL 2007;172(2-3):179. 

644. Hurley JM, West JB, Ehleringer JR. Tracing retail cannabis in the United 

States: Geographic origin and cultivation patterns. INTERNATIONAL 

JOURNAL OF DRUG POLICY 2010;21(3):222-228. 

645. Shibuya EK, Sarkis JES, Negrini-Neto O, Ometto JPHB. Multivariate 

classification based on chemical and stable isotopic profiles in sourcing 

the origin of marijuana samples seized in Brazil. JOURNAL OF THE 

BRAZILIAN CHEMICAL SOCIETY 2007;18(1):205-214. 

646. West JB, Hurley JM, Ehleringer JR. Stable Isotope Ratios of Marijuana. I. 

Carbon and Nitrogen Stable Isotopes Describe Growth Conditions. 


647. West JB, Hurley JM, Dudas FO, Ehleringer JR. The stable isotope ratios 

of marijuana. II. Strontium isotopes relate to geographic origin. JOURNAL 

OF FORENSIC SCIENCES 2009;54(6):1261-1269. 

648. Zacca JJ, Fava N, Santos RV, Mancini LH, Bastos MQ, Pacheco WL, 

Lima BA. Stable Isotope Ratios of Marijuana from Brazil and Paraguay: 

Carbon and Nitrogen as Guides for Growth Conditions. The Fourth 


796

5.5 Methamphetamine: 

649. Billault I, Courant F, Pasquereau L, Derrien S, Robins RJ, Naulet N. 

Correlation between the synthetic origin of methamphetamine samples 

and their N-15 and C-13 stable isotope ratios. ANALYTICA CHIMICA 

ACTA 2007;593(1):20-29. 

650. Collins M, Cawley AT, Heagney AC, Kissane L, Robertson J, Salouros H. 

delta C-13, delta N-15 and delta H-2 isotope ratio mass spectrometry 

of ephedrine and pseudoephedrine: application to methylamphetamine 

profiling. RAPID COMMUNICATIONS IN MASS SPECTROMETRY 

2009;23(13):2003-2010. 

651. Cox M, Klass G, Koo CWM. Manufacturing by-products from, and 

stereochemical outcomes of the biotransformation of benzaldehyde used 

in the synthesis of methamphetamine. FORENSIC SCIENCE 


652. David GE, Hibbert DB, Frew RD, Hayman AR. Significant determinants of 

isotope composition during hi/p-red synthesis of methamphetamine. 

AUSTRALIAN JOURNAL OF CHEMISTRY 2010;63(1):22-29. 

653. Dujourdy L, Dufey V, Besacier F, Miano N, Marquis R, Lock E, Aalberg L, 

Dieckmann S, Zrcek F, Bozenko JS. Drug intelligence based on organic 

impurities in illicit MA samples. FORENSIC SCIENCE INTERNATIONAL 

2008;177(2-3):153-161. 

654. Ikehara Y, Kurashima N, Makino Y, Nagano T, Sanuki K, Urano Y. Use of 

stable isotope ratios for profiling of industrial ephedrine samples: 

Application of hydrogen isotope ratios in combination with carbon and 

nitrogen. FORENSIC SCIENCE INTERNATIONAL 2009;189(1-3):14-18. 

655. Inoue H, Iwata YT, Kuwayama K. Characterization and profiling of 

methamphetamine seizures. JOURNAL OF HEALTH SCIENCE 

2008;54(6):615-622. 

656. Iwata YT, Kuwayama K, Tsujikawa K, Miyaguchi H, Kanamori T, Inoue H. 

Evaluation method for linking methamphetamine seizures using stable 

carbon and nitrogen isotopic compositions: A complementary study with 

impurity profiling. RAPID COMMUNICATIONS IN MASS 

SPECTROMETRY 2008;22(23):3816-3822. 

657. Ko BJ, Suh SI, Suh YJ, In MK, Kim SH. The impurity characteristics of 

methamphetamine synthesized by Emde and Nagai method. FORENSIC 

SCIENCE INTERNATIONAL 2007;170(2-3):142. 

658. Kunalan V, Nic Daeid N, Kerr WJ, Buchanan HAS, McPherson AR. 

Characterization of route specific impurities found in methamphetamine 

synthesized by the leuckart and reductive amination methods. 

ANALYTICAL CHEMISTRY 2009;81(17):7342-7348. 

659. Kurashima N, Makino Y, Urano Y, Sanuki K, Ikehara Y, Nagano T. Use of 

stable isotope ratios for profiling of industrial ephedrine samples: 

Application of hydrogen isotope ratios in combination with carbon and 

nitrogen. FORENSIC SCIENCE INTERNATIONAL 2009;189(1-3):14-18. 

797

660. Kuwayama K, Inoue H, Kanamori T, Tsujikawa K, Miyaguchi H, Iwata Y, 

Kamo N, Kishi T. Contribution of thermal desorption and liquid-liquid 

extraction for identification and profiling of impurities in methamphetamine 

by gas chromatography-mass spectrometry. FORENSIC SCIENCE 

INTERNATIONAL 2007;171(1):9-15. 

661. Kuwayama K, Inoue H, Phorachata J, Kongpatnitiroj K, Puthaviriyakorn V, 

Tsujikawa K, Miyaguchi H, Kanamori T, Iwata YT, Kamo N, Kishi T. 

Comparison and classification of methamphetamine seized in Japan and 

Thailand using gas chromatography with liquid-liquid extraction and solidphase 

microextraction. FORENSIC SCIENCE INTERNATIONAL 

2008;175(2-3):85-92. 

662. Lee JS, Chung HS, Kuwayama K, Inoue H, Lee MY, Park JH. 

Determination of impurities in illicit methamphetamine seized in Korea and 

Japan. ANALYTICA CHIMICA ACTA 2008;619(1):20-25. 

663. Lee JS, Yang WK, Han EY, Lee SY, Park YH, Lim MA, Chung HS, Park 

JH. Monitoring precursor chemicals of methamphetamine through 

enantiomer profiling. FORENSIC SCIENCE INTERNATIONAL 

2007;173(1):68. 

664. Li Q, Xu H, Song Z, Li Q. Analysis of organic impurity profiling of illicit 

methamphetamine bydrochloride crystal by GC-MS. FENXI CESHI 

XUEBAO 2007;26(3):310-314. 

665. Matsumoto T, Urano Y, Makino Y, Kikura-Hanajiri R, Kawahara N, Goda 

Y, Nagano T. Evaluation of characteristic deuterium distributions of 

ephedrines and methamphetamines by NMR spectroscopy for drug 

profiling. ANALYTICAL CHEMISTRY 2008;80(4):1176-1181. 

666. Person EC, Golden ML. Current views on the molecular structures of 

phosphorus allotropes. JOURNAL OF THE CLANDESTINE 


2008;19(4):20. 

667. Person E, Heegel RA, Knops LA, Northrop DM. Phosphorus-containing 

reducing agents: a review of their chemistry and use in the manufacture of 

methamphetamine and the significance of observed phosphate, phosphite, 

and hypophosphite in clandestine laboratory casework. JOURNAL OF 


ASSOCIATION 2008;18(2):7-38. 

668. Qi Y, Evans I, McCluskey A. New impurity profiles of recent Australian 

imported “Ice”: Methamphetamine impurity profiling and the identification 

of (pseudo)ephedrine and Leuckart specific marker compounds. 

FORENSIC SCIENCE INTERNATIONAL 2007;169(2-3):173. 

669. Salouros H, Collins M, George AV, Davies S. Isolation and identification of 

three by-products found in methylamphetamine synthesized by the emde 

route. JOURNAL OF FORENSIC SCIENCES 2010;55(3):605-615. 

798

670. Schneiders S, Holdermann T, Dahlenburg R. Comparative analysis of 1phenyl-2-propanone 

(P2P), an amphetamine-type stimulant precursor, 

using stable isotope ratio mass spectrometry. Presented in part as a 

poster at the 2nd meeting of the Joint European Stable Isotope User 

Meeting (JESIUM), Giens, France, September 2008. SCIENCE & 

JUSTICE 2009;49(2):94-101. 

671. van Breukelen M, Hoitink M, Poortman A. Comparison of p2p seizures 

using GC/MS impurity profiling and IRMS. The Fourth Forensic Isotope 

Ratio Mass Spectrometry Conference: 2010. 

672. Zhang J, Zhang D, Han X, Qiao J, Yang S, Xin G. Study on the linked 

samples of methamphetamine identified by the impurity profile. 

WEISHENG YANJIU 2008;37(6):736-739, 744. 

673. Zhang J, Zhang D, Han X, Yang S, Xin G. Determination of the synthetic 

route based on impurity profiling of methamphetamine. WEISHENG 

YANJIU 2008;37(6):740-744. 

674. Zhang J, Zhang D, Zhang W, Qiao J, Yang S, Xin G. Monitoring precursor 

chemicals through enantiomer profiling of methamphetamine. WEISHENG 

YANJIU 2009;38(4):426-428. 

675. Zhang JX, Zhang DM, Han XG. Identification of impurities and statistical 

classification of methamphetamine hydrochloride drugs seized in China. 


5.6 3,4-Methylenedioxymethamphetamine: 

676. Baer I, Gurny R, Margot P. NIR analysis of cellulose and lactose: 

Application to ecstasy tablet analysis. FORENSIC SCIENCE 


677. Bolck A, Weyermann C, Dujourdy L, Esseiva P, van den Berg J. Different 

likelihood ratio approaches to evaluate the strength of evidence of MDMA 

tablet comparisons. FORENSIC SCIENCE INTERNATIONAL 2009;191(1- 

3):42-51. 

678. Bonadio F, Margot P, Delemont O, Esseiva P. Optimization of HS- 

SPME/GC-MS analysis and its use in the profiling of illicit ecstasy tablets 

(Part 1). FORENSIC SCIENCE INTERNATIONAL 2009;187(1-3):73-80. 

679. Bonadio F, Margot P, Delemont O, Esseiva P. Headspace solid-phase 

microextraction (HS-SPME) and liquid-liquid extraction (LLE): Comparison 

of the performance in classification of ecstasy tablets (Part 2). FORENSIC 


680. Buchanan HAS, Nic Daeid N, Meier-Augenstein W, Kemp HF, Kerr 

WJ, Middleditch M. Emerging use of isotope ratio mass spectrometry as a 

tool for discrimination of 3,4-methylenedioxymethamphetamine by 

synthetic route. ANALYTICAL CHEMISTRY 2008;80(9):3350-3356. 

681. Byrska B, Zuba D. Profiling of 3,4-methylenedioxymethamphetamine by 

means of high-performance liquid chromatography. ANALYTICAL & 


799

682. Cioroch K, Zuba D. Analytical approaches used for profiling of ecstasy 

tablets. Z ZAGADNIEN NAUK SADOWYCH 2007;69:71-89. 

683. Cox M, Klass G, Morey S, Pigou P. Chemical markers from the peracid 

oxidation of isosafrole. FORENSIC SCIENCE INTERNATIONAL 

2008;179(1):44-53. 

684. Fierro I, Deban L, Pardo R, Tascon M, Vazquez D. Analysis of heavy 

metals in ecstasy tablets by electrochemical methods. TOXICOLOGICAL 

AND ENVIRONMENTAL CHEMISTRY 2007;89(3):411. 

685. Kochana J, Tomaszewski W, Moszczynski T, Zakrzewska A, Parczewski 

A. Application of carbon adsorbents for extraction of MDMA impurities in 

TLC drug profiling. JOURNAL OF LIQUID CHROMATOGRAPHY & 

RELATED TECHNOLOGIES 2008;31(6):819-827. 

686. Koper C, van den Boom C, Wiarda W, Schrader M, de Joode P, van der 

Peijl G, Boick A. Elemental analysis of 3,4methylenedioxymethamphetamine 

(MDMA): A tool to determine the 

synthesis method and trace links. FORENSIC SCIENCE 


687. Marquis R, Weyermann C, Delaporte C, Esseiva P, Aalberg L, Besacier F, 

Bozenko JS, Dahlenburg R, Kopper C, Zrcek F. Drug intelligence based 

on MDMA tablets data 2. Physical characteristics profiling. FORENSIC 


688. Meisinger SC, Waddell-Smith R. Comparison of extraction procedures for 

organic impurity profiling of seized MDMA tablets. 2008 American 

Academy of Forensic Sciences Annual Meeting. 

689. Meng PJ, Margot P. Constituent profiling of ecstasy seizures by GC-MS 

after joint derivatization with MSTFA and MBTFA. FORENSIC 

TOXICOLOGY 2010;28(1):52-54. 

690. Milliet Q, Weyermann C, Esseiva P. The profiling of MDMA tablets: A 

study of the combination of physical characteristics and organic impurities 

as sources of information. FORENSIC SCIENCE INTERNATIONAL 

2009;187(1-3):58-65. 

691. Waddell-Smith RJH. A review of recent advances in impurity profiling of 

illicit MDMA samples. JOURNAL OF FORENSIC SCIENCES 

2007;52(6):1297-1304. 

692. Weyermann C, Marquis R, Delaporte C, Esseiva P, Lock E, Aalberg 

L, Bozenko JS, Dieckmann S, Dujourdy L, Zrcek F. Drug intelligence 

based on MDMA tablets data - I. Organic impurities profiling. FORENSIC 


5.7 Multi-Drug and Miscellaneous: 

693. Gross KC. Passive tagging of pharmaceutical products with noble gas 

isotopes. (Patent) CHEMICAL ABSTRACTS 2007;147:39338v. 

694. Jasper JP, Weaner LE, Hayes JM. Process patent protection: 

Characterizing synthetic pathways by stable-isotopic measurements. 

PHARMACEUTICAL TECHNOLOGY 2007;31(3):68-78. 

800

695. Lock CM, Meier-Augenstein W. Investigation of isotopic linkage between 

precursor and product in the synthesis of a high explosive. FORENSIC 


696. Marclay F, Pazos D, Delemont O, Esseiva P, Saudan C. Potential of IRMS 

technology for tracing gamma-butyrolactone (GBL). FORENSIC SCIENCE 


697. Wise SH, Almirall JR. Chemical taggant detection and analysis by laserinduced 

breakdown spectroscopy. APPLIED OPTICS 2008;47(31):G15- 

G20. 

698. Yip YC, Lam JCW, Tong WF. Applications of lead isotope ratio 

measurements. TrAC-TRENDS IN ANALYTICAL CHEMISTRY 

2008;27(5):460-480. 

699. Zhang G, Jin Y. Recent advances on the techniques of fluorescentlabeling 

and its application. WUJING YIZUEYUAN ZUEBAO 

2008;17(6):550-552. 

700. Zhu EY, Lin Y, Zhuang ZY. Partial least squares variable selection method 

and its application in drug source analysis. CHINESE JOURNAL OF 

ANALYTICAL CHEMISTRY 2007;35:973-977. 

6. Analysis of Non-Controlled Pharmaceuticals, Pseudo-Drugs, 

Issue: 

Adulterants, Diluents, and Precursors 

Most "street-level" drugs are "cut" with various adulterants and diluents. Many of 

these cutting agents are pharmaceutical products or precursors. Others are 

“carry-through” compounds present in precursors (especially in cold remedy 

products). Separation and identification of these extraneous materials can be 

tedious, especially in exhibits which contain many components. In addition, new 

or unusual adulterants and/or diluents are occasionally identified in drug exhibits, 

and standard analytical data are required for these substances. Finally, 

improved methods of analysis, i.e., faster, more discriminatory, less costly, etc., 

are needed for all cutting agents. 

6.1 Ephedra, Ephedrine, and/or Pseudoephedrine and Related 

Compounds: 

701. Ansell RJ, Wang DY, Kuah JKL. Imprinted polymers for chiral resolution of 

(+/-)-ephedrine. Part 2: Probing pre-polymerisation equilibria in different 

solvents by NMR. ANALYST 2008;133(12):1673-1683. 

702. Ansell RJ, Wang DY. Imprinted polymers for chiral resolution of (+/-)ephedrine. 

Part 3: NMR predictions and HPLC results with alternative 

functional monomers. ANALYST 2009;134(3):564-576. 

801

703. bin Abdullah AFL, Miskelly GM. Formation of trifluoroacetylated ephedrine 

during the analysis of a pseudoephedrine-formaldehyde adduct by TFAA 

derivatization followed by GC-MS. JOURNAL OF FORENSIC SCIENCES 

2009;54(2):365-369. 

704. Boff BD, Sebben VC, Paliosa PK, Azambuja I, Singer RB, Limberger RP. 

Investigation of the presence of ephedrines in Ephedra tweediana Fisch & 

CA Meyer and E. triandra Tul. (Ephedraceae) collected in Porto 

Alegre/RS. REVISTA BRASILEIRA DE FARMACOGNOSIA-BRAZILIAN 

JOURNAL OF PHARMACOGNOSY 2008;18(3):394-401. 

705. Fan Q, Wang YL, Sun P, Liu S, Li Y. Discrimination of Ephedra plants with 

diffuse reflectance FT-NIRS and multivariate analysis. TALANTA 

2010;80(3):1245-1250. 

706. Ghafoor S, Shah MM, Ahmad H, Swati ZA, Shah SH, Pervez A, Farooq U. 

Molecular characterization of Ephedra species found in Pakistan. 

GENETICS AND MOLECULAR RESEARCH 2007;6(4):1123-1130. 

707. Guerreiro AR, Korkhov V, Mijangos I, Piletska EV, Rodins J, Turner APF, 

Piletsky SA. Influence of continuous magnetic field on the separation of 

ephedrine enantiomers by molecularly imprinted polymers. BIOSENSORS 

& BIOELECTRONICS 2008;23(7):1189-1194. 

708. Hakey PM, Allis DG, Hudson MR, Ouellette W, Korter TM. Investigation of 

(1R,2S)-(-)-ephedrine by cryogenic terahertz spectroscopy and solid-state 

density functional theory. CHEMPHYSCHEM 2009;10(14):2434-2444. 

709. Hayashi K, Shimura K, Makino T, Mizukami H. Comparison of the 

contents of kampo decoctions containing ephedra herb when prepared 

simply or by re-boiling according to the traditional theory. JOURNAL OF 

NATURAL MEDICINES 2010;64(1):70-74. 

710. Inoko A, Kakiuchi N, Yoshimitsu M, Cai SQ, Mikage M. Ephedra resource 

in sichuan and yunnan provinces 2007. BIOLOGICAL & 

PHARMACEUTICAL BULLETIN 2009;32(9):1621-1623. 

711. Kakiuchi N, Inoue K, Kurita Y, Ohkubo K, Tsuda Y, Mikage M. Survey of 

Ephedra resources in the northern areas of Pakistan and their genetic 

diversity. JOURNAL OF NATURAL MEDICINES 2007;61(3):357-365. 

712. Kitani Y, Zhu S, Omote T, Tanaka K, Batkhuu J, Sanchir C, Fushimi H, 

Mikage M, Komatsu K. Molecular analysis and chemical evaluation of 

ephedra plants in mongolia. BIOLOGICAL & PHARMACEUTICAL 

BULLETIN 2009;32(7):1235-1243. 

713. Lu J, Jin X, Mao PH, Ma XD, Ling HQ, Fan YH, Yu L, Wu BS, Ouyang PK. 

Transfer of ephedra genomic DNA to yeasts by ion implantation. APPLIED 

BIOCHEMISTRY AND BIOTECHNOLOGY 2009;158(3):571-581. 

714. Mao P, Ma X, Jin X, Yang H, Lou K. Production of l-ephedrine and dpseudoephedrine 

in recombined yeasts obtained by Argon ion 

implantation mediated Ephedra genome DNA transformation. 

WEISHENGWU XUEBAO 2007;47(5):905-909. 

802

715. Okada T, Nakamura Y, Kanaya S, Takano A, Malla KJ, Nakane 

T, Kitayama M, Sekita S. Metabolome analysis of ephedra plants with 

different contents of ephedrine alkaloids by using UPLC-Q-TOF-MS. 

PLANTA MEDICA 2009;75(12):1356-1362. 

716. Pellati F, Benvenuti S. Determination of ephedrine alkaloids in Ephedra 

natural products using HPLC on a pentafluorophenylpropyl stationary 

phase. JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL 

ANALYSIS 2008;48(2):254-263. 

717. Ranieri TL, Ciolino LA. Rapid selective screening and determination of 

ephedrine alkaloids using GC-MS footnote mark. PHYTOCHEMICAL 

ANALYSIS 2008;19(2):127-135. 

718. Valente AJM, Ribeiro ACF, Marques JMC, Abreu PE, Lobo VMM, Kataky 

R. Transport properties of aqueous solutions of (1R,2S)-(-)- and (1S,2R)- 

(+)-ephedrine hydrochloride at different temperatures. JOURNAL OF 

CHEMICAL AND ENGINEERING DATA 2010;55(3):1145-1152. 

719. Wang LL, Kakiuchi N, Mikage M. Studies of Ephedra plants in Asia. Part 

6: Geographical changes of anatomical features and alkaloids content of 

Ephedra sinica. JOURNAL OF NATURAL MEDICINES 2010;64(1):63-69. 

720. Wang LS, Zhao DQ, Liu YH. GC-MS analysis of the supercritical CO2 fluid 

extraction of Ephedra sinica roots and its antisudorific activity. 

CHEMISTRY OF NATURAL COMPOUNDS 2009;45(3):434-436. 

721. Wilhalm T. Current survey of Ephedra stands (Ephedra helvetica, 

Ephedraceae, Gnetales) in South Tyrol. GREDLERIANA 2007;7:69-90. 

722. Zhao W, Deng AJ, Du GH, Zhang JL, Li ZH, Qin HL. Chemical 

constituents of the stems of Ephedra sinica. JOURNAL OF ASIAN 

NATURAL PRODUCTS RESEARCH 2009;11(2):168-171. 

723. Zhou L, Zhou XM, Luo Z, Wang WP, Yan N, Hu ZD. In-capillary 

derivatization and analysis of ephedrine and pseudoephedrine by micellar 

electrokinetic chromatography with laser-induced fluorescence detection. 

JOURNAL OF CHROMATOGRAPHY A 2008;1190(1-2):383-389. 

6.2 Phenylpropanolamine: 

724. Abbasi K, Bhanger MI, Khuhawar MY. Capillary-gas chromatographic 

determination of phenylpropanolamine using acetylacetone as derivatizing 

reagent in pharmaceutical preparations. JOURNAL OF THE CHEMICAL 

SOCIETY OF PAKISTAN 2008;30(5):692-695. 

725. Hadad GM, El-Gindy A, Mahmoud WMM. Development and validation of 

chemometrics-assisted spectrophotometry and liquid chromatography 

methods for the simultaneous determination of the active ingredients in 

two multicomponent mixtures containing chlorpheniramine maleate and 

phenylpropanolamine hydrochloride. JOURNAL OF AOAC 


803

6.3 Other Adulterants/Diluents (including mixtures containing Ephedrine 

and/or Pseudoephedrine): 

726. Amer SM, Abbas SS, Shehata MA, Ali NM. Simultaneous determination of 

phenylephrine hydrochloride, guaifenesin, and chlorpheniramine maleate 

in cough syrup by gradient liquid chromatography. JOURNAL OF AOAC 


727. Amin AS, Dessouki HA, Moustafa MM, Ghoname MS. Spectrophotometric 

methods for sertraline hydrochloride and/or clidinium bromide 

determination in bulk and pharmaceutical preparations. CHEMICAL 

PAPERS 2009;63(6):716-722. 

728. Brettell TA. Chiral analysis of methorphan and citalopram using HPLC. 

2008 American Academy of Forensic Sciences Annual Meeting. 

729. El-Naby, EH. Polymeric membrance sensors for the selective 

determination of dextromethorphan in pharmaceutical preparations. 


730. Liao CZ, Nicklaus MC. Comparison of nine programs predicting pK(a) 

values of pharmaceutical substances. JOURNAL OF CHEMICAL 

INFORMATION AND MODELING 2009;49(12):2801-2812. 

731. Lin MC, Liu YC, Lin YL, Lin JH. Identification of a tadalafil analogue 

adulterated in a dietary supplement. JOURNAL OF FOOD AND DRUG 

ANALYSIS 2009;17(6):451-458. 

732. Liu Y, Ge H, Zhao K, Yu L. Determination of three chemical components 

added illegally in a analgesia traditional Chinese medicine preparation by 

TLC-MS. ZHONGGUO YAOXUE ZAZHI 2008;43(22):1747-1750. 

733. McGehee MC. Etodolac: An analytical profile. MICROGRAM JOURNAL 

2008;6(3-4):104. 

734. Pavlic M, Schubert B, Libiseller K, Oberacher H. Comprehensive 

identification of active compounds in tablets by flow-injection datadependent 

tandem mass spectrometry combined with library search. 


735. Rodriguez N, Vidot JV, Panelli J, Colon H, Ritchie B, Yamamura Y. GC- 

MS confirmation of xylazine (Rompun), a veterinary sedative, 

in exchanged needles. DRUG AND ALCOHOL DEPENDENCE 

2008;96(3):290-293. 

6.4 Theophylline: 

736. Zare-Shahabadi V, Shamsipur M, Hemmatenejad B, Akhond M. 

Simultaneous determination of guaifenesin and theophylline by 

chemometrics methods. ANALYTICAL LETTERS 2010;43(4):687-700. 

804


737. Gratz SR, Zeller M, Mincey DW, Flurer CL. Structural characertization of 

sulfoaildenafil, an analog of sildenafil. JOURNAL OF PHARMACEUTICAL 

AND BIOMEDICAL ANALYSIS 2009;50(2):228-231. 

738. Jouyban A, Soltanpour S, Tamizi E. Solubility prediction of solutes in 

aqueous mixtures of ethylene glycols. PHARMAZIE 2008;63(7):548-550. 

739. Kamb V. Analytical profile of lisdexamfetamine dimesylate (Vyvanse). 



740. Sakar MK, Alper-Yegul N. Isolation and identification of N-monodesmethylsibutramine 

in a slimming herbal product. ACTA 

PHARMACEUTICA SCIENCIA 2008;50(2):97-100. 

7. New and/or Improved Instrumental Techniques 

Issue: 

Forensic Chemists must maintain familiarity with updates in current instrumental 

techniques and become versant in new, improved methods of analysis. 

Solution: 

Improved/existing and new technologies are reviewed and applied to both routine 

and specialized analyses of drugs. In cases where improved performance is 

observed, case reports are generated for the forensic community. 

7.1 Capillary Electrophoresis (and Related Techniques, including 

Tandem Techniques): 

741. Assuncao NA, Bechara EJH, Simionato AVC, Tavares MFM, Carrilho E. 

Capillary electrophoresis coupled to mass spectrometry (CE-MS): Twenty 

years of development. QUIMICA NOVA 2008;31(8):2124-2133. 

742. Aturki Z, D'Orazio G, Fanali S, Rocco A, Bortolotti F, Gottardo R, Tagliaro 

F. Capillary electrochromatographic separation of illicit drugs employing a 

cyano stationary phase. JOURNAL OF CHROMATOGRAPHY A 

2009;1216(17):3652-3659. 

743. Breadmore MC, Thabano JRE, Dawod M, Kazarian AA, Quirino JP, Guijt 

RM. Recent advances in enhancing the sensitivity of electrophoresis and 

electrochromatography in capillaries and microchips (2006-2008). 

ELECTROPHORESIS 2009;30(1):230-248. 

805

744. Du Y, Wang E. Separation and detection of narcotic drugs on a microchip 

using micellar electrokinetic chromatography and 

electrochemiluminescence. ELECTROANALYSIS 2008;20(6):643-647. 

745. Ganzera M. Quality control of herbal medicines by capillary 

electrophoresis: Potential, requirements and applications. 


746. Greenspoon SA, Yeung SHI, Johnson KR, Chu WK, Rhee HN, 

McGuckian AB, Crouse CA, Chiesl TN, Barron AE, Scherer JR, Ban JD, 

Mathies RA. A forensic laboratory tests the Berkeley microfabricated 

capillary array electrophoresis device. JOURNAL OF FORENSIC 

SCIENCES 2008;53(4):828-837. 

747. Gubitz G, Schmid MG. Chiral separation by capillary electromigration 

techniques. JOURNAL OF CHROMATOGRAPHY A 2008;1204(2):140- 

156. 

748. Hoeman KW, Culbertson CT. A novel, environmentally friendly sodium 

lauryl ether sulfate-, cocamidopropyl betaine-, cocamide 

monoethanolamine-containing buffer for MEKC on microfluidic devices. 


749. Huck CW, Huck-Pezzei V, Bakry R, Bachmann S, Najam-ul-Haq M, 

Rainer M, Bonn GK. Capillary electrophoresis coupled to mass 

spectrometry for forensic analysis. OPEN CHEMICAL ENGINEERING 

JOURNAL 2007;1:30-43. 

750. Huhn C, Puetz M, Holthausen I, Pyell U. Separation of very hydrophobic 

analytes by micellar electrokinetic chromatography. I. Optimization of the 

composition of the sample solution for the determination of the aromatic 

ingredients of sassafras and other essential oils of forensic interest. 


751. Jiang ZJ, Thorogate R, Smith NW. Highlighting the role of the hydroxyl 

position on the alkyl spacer of hydroxypropyl-beta-cyclodextrin for 

enantioseparation in capillary electrophoresis. JOURNAL OF 

SEPARATION SCIENCE 2008;31(1):177-187. 

752. Jouyban A, Kenndler E. Impurity analysis of pharmaceuticals using 

capillary electromigration methods. ELECTROPHORESIS 

2008;29(17):3531-3551. 

753. Li MJ, Zhou JY, Gu X, Wang Y, Huang XJ, Yan C. Quantitative capillary 

electrophoresis and its application in analysis of alkaloids in tea, coffee, 

coca cola, and theophylline tablets. JOURNAL OF SEPARATION 

SCIENCE 2009;32(2):267-274. 

754. Loden H, Pettersson C, Arvidsson T, Amini A. Quantitative determination 

of salbutamol in tablets by multiple-injection capillary zone 

electrophoresis. JOURNAL OF CHROMATOGRAPHY A 2008;1207(1- 

2):181-185. 

755. Magana JJ, de la Luz Arenas-Sordo M, Gomez R. Capillary 

electrophoresis, a new diagnostic tool. REVISTA MEDICA DE CHILE 

2009;137(7):946-956. 

806

756. Mala Z, Slampova A, Gebauer P, Bocek P. Contemporary sample 

stacking in CE. ELECTROPHORESIS 2009;30(1):215-229. 

757. Preinerstorfer B, Lammerhofer M, Lindner W. Advances in 

enantioselective separations using electromigration capillary techniques. 


758. Rohrbasser C, Rheme D, Decastel S, Roth S, Aja Montes M, Veuthey J, 

Rudaz S. A new capillary electrophoresis device with deep UV detector 

based on LED technology. CHIMIA 2009;63(12):890-891. 

759. Saavedra L, Nickerson B, Borjas RE, Lynen F, Sandra P. 

Enantioseparation of pharmaceutical compounds by multiplexed 

capillary electrophoresis using highly sulphated alpha-, beta- and gammacyclodextrins. 

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL 

TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES 

2008;875(1):248-253. 

760. Scriba GKE. Cyclodextrins in capillary electrophoresis enantioseparations 

- Recent developments and applications. JOURNAL OF SEPARATION 

SCIENCE 2008;31(11):1991-2011. 

761. Staub A, Giraud S, Saugy M, Rudaz S, Veuthey J, Schappler J. CE-ESI- 

TOF/MS for human growth hormone analysis. ELECTROPHORESIS 

2010;31(2):388-395. 

762. Tagliaro F, Bortolotti F. Recent advances in the applications of CE to 

forensic sciences (2005-2007). ELECTROPHORESIS 2008;29(1):260- 

268. 

763. Tagliaro F, Pascali J, Fanigliulo A, Bortolotti F. Recent advances in the 

application of CE to forensic sciences: An update over years 2007-2009. 

ELECTROPHORESIS 2010;31(1):251-259. 

764. Yang YL, Zhu CF, Shen J, Hao AY. Enantioseparation in capillary 

electrophoresis using 6-oligo(lactic acid)cyclomaltoheptaose as a chiral 

selector. ANALYTICAL SCIENCES 2009;25(11):1315-1318. 

765. Yin DG, Wu MH, Xie CJ, Zhang L, Liu BH, Zhou XZ, Wang P. A Novel 

Capillary Electrophoresis-Chemiluminescence system for separation and 

detection of amino acids. CHINESE JOURNAL OF ANALYTICAL 

CHEMISTRY 2009;37(1):152-156. 

766. Zhu FX, Du YX, Chen JQ, Chen B, Zhu YQ, Zhai X, Xu S, Zhou WH. 

Enantioselective separation of basic drugs by CE with polygalacturonic 

acid as a novel chiral selector. CHROMATOGRAPHIA 2009;69(11- 

12):1315-1320. 

7.2 Extraction Techniques: 

767. Almirall J, Perr J, Guerra P. Method and apparatus for extraction, 

detection, and characterization of vapors from explosives, taggants in 

explosives, controlled substances, and biohazards. APPLICATION: US 

2008-135954;9 June 2008. 

768. Brewer W. Pipette tips for extraction, sample collection and sample 

cleanup, and methods for their use. PCT Int. Appl. WO 2008 103,828. 

807

769. Gonzalez-Marino I, Quintana JB, Rodriguez I, Cela R. Determination of 

drugs of abuse in water by solid-phase extraction, derivatisation and gas 

chromatography-ion trap-tandem mass spectrometry. JOURNAL OF 

CHROMATOGRAPHY A 2010;1217(11):1748-1760. 

770. Juhascik MP, Jenkins AJ. Comparison of liquid/liquid and solid-phase 

extraction for alkaline drugs. JOURNAL OF CHROMATOGRAPHIC 

SCIENCE 2009;47(7):553-557. 

771. Kmecz I, Simandi B, Szekely E, Lovasz J, Fogassy E. Application of 

mixtures of tartaric acid derivatives in resolution via supercritical fluid 

extraction. CHIRALITY 2007;19(6):430-433. 

772. Lu J, Shen D, Liu Y. Solubilization of microemulsion to indissolvable drugs 

and its clinical application. WUJING YIXUEYUAN XUEBAO 

2008;17(3):241-243. 

773. Madej K. Microwave-assisted and cloud-point extraction in determination 

of drugs and other bioactive compounds. TrAC - TRENDS IN 


774. Person EC, Sunderson NS. Liquid-liquid extractions of phenylephrine. 



775. Wietecha-Posluszny R, Wozniakiewicz M, Garbacik A, Koscielniak P. 

Application of microwave-assisted extraction in isolation of psychotropic 

drugs from biological material. Z ZAGADNIEN NAUK SADOWYCH 

2007;70:187-197. 

776. Wille SMR, Lambert WEE. Recent developments in extraction procedures 

relevant to analytical toxicology. ANALYTICAL AND BIOANALYTICAL 

CHEMISTRY 2007;388(7):1381-1391. 

777. Xu L, Basheer C, Lee HK. Chemical reactions in liquid-phase 

microextraction. JOURNAL OF CHROMATOGRAPHY A 

2009;1216(4):701-707. 

7.3 Gas Chromatography (and Tandem GC Techniques): 

778. Ali S, Ashraf-Khorassani M, Taylor LT, Agah M. MEMS-based semipacked 

gas chromatography columns. SENSORS AND ACTUATORS B: 

CHEMICAL 2009;141(1):309-315. 

779. Binkley J. Rapid analysis of drugs by abuse by gas chromatography-time 

of flight mass spectrometry. Mid-Atlantic Association of Forensic Scientists 


780. Chu FL, Yaylayan VA. Model studies on the oxygen-induced formation of 

benzaldehyde from phenylacetaldehyde using pyrolysis GC-MS and FTIR. 

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 

2008;56(22):10697-10704. 

781. Damm M, Rechberger G, Kollroser M, Kappe CO. An evaluation of 

microwave-assisted derivatization procedures using hyphenated mass 

spectrometric techniques. JOURNAL OF CHROMATOGRAPHY A 

2009;1216(31):5875-5881. 

808

782. Fan WZ, Zhang Y, Carr PW, Rutan SC, Dumarey M, Schellinger AP, Pritts 

W. Application of Snyder-Dolan classification scheme to the selection of 

"orthogonal" columns for fast screening of illicit drugs and impurity profiling 

of pharmaceuticals - I. Isocratic elution. JOURNAL OF 


783. Fialkov AB, Gordin A, Amirav A. Hydrocarbons and fuels analyses with the 

supersonic gas chromatography mass spectrometry-The novel concept of 

isomer abundance analysis. JOURNAL OF CHROMATOGRAPHY A 

2008;1195(1-2):127-135. 

784. Grigor'ev AM, Savchuk SA. Correlation of the parameters of screening 

libraries of the gas-chromatographic retention data. JOURNAL OF 

ANALYTICAL CHEMISTRY 2010;65(4):388-397. 

785. Groger T, Schaffer M, Putz M, Ahrens B, Drew K, Eschner 

M, Zimmermann R. Application of two-dimensional gas chromatography 

combined with pixel-based chemometric processing for the chemical 

profiling of illicit drug samples. JOURNAL OF CHROMATOGRAPHY A 

2008;1200(1):8-16. 

786. Hadef Y, Kaloustian J, Nicolay A, Portugal H. Thermal stability evaluation 

of doping compounds before GC-MS analysis by DSC. JOURNAL OF 

THERMAL ANALYSIS AND CALORIMETRY 2008;93(2):553-560. 

787. Hoggard JC, Wahl JH, Synovec RE, Mong GM, Fraga CG. Impurity 

profiling of a chemical weapon precursor for possible forensic signatures 

by comprehensive two-dimensional gas chromatography/mass 

spectrometry and chemometrics. ANALYTICAL CHEMISTRY 

2010;82(2):689-698. 

788. Kirkpatrick DS. A practical guide to drug identification using fast GC/MS. 


789. Kudo K, Ishida T, Hikiji W, Hayashida M, Uekusa K, Usumoto Y, Tsuji A, 

Ikeda N. Construction of calibration-locking databases for rapid and 

reliable drug screening by gas chromatography-mass spectrometry. 

FORENSIC TOXICOLOGY 2009;27(1):21-31. 

790. Kudo K, Ishida T, Ikeda N. Development of a systematic screening 

procedure for abused drugs without using standard compounds by gas 

chromatography/mass spectrometry. JOURNAL OF THE MASS 

SPECTROMETRY SOCIETY OF JAPAN 2008;56(3):123-30. 

791. Lin DL, Wang SM, Wu CH, Chen BG, Liu RH. Chemical derivatization for 

the analysis of drugs by GC-MS. YAOWU SHIPIN FENXI 2008;16(1):1-10. 

792. Maurer HH. Forensic screening with GC-MS. HANDBOOK OF 

ANALYTICAL SEPARATIONS 2008;6:425-445. 

793. Meier-Augenstein W, NicDaeid N. Feasibility of source identification of 

seized street drugs samples by exploiting differences in isotopic 

composition at natural abundance level by GC/MS as compared to isotope 

ratio mass spectrometry (IRMS). FORENSIC SCIENCE INTERNATIONAL 

2008;174(2-3):259-261. 

809

794. Mitrevski BS, Wilairat P, Marriott PJ. Comprehensive two-dimensional gas 

chromatography improves separation and identification of anabolic agents 

in doping control. JOURNAL OF CHROMATOGRAPHY A 

2010;1217(1):127-135. 

795. Ojanpera I, Rasanen I. Forensic screening by gas chromatography. 

HANDBOOK OF ANALYTICAL SEPARATIONS 2008;6:403-424. 

796. Schmarr HG, Bernhardt J. Profiling analysis of volatile compounds from 

fruits using comprehensive two-dimensional gas chromatography and 

image processing techniques. JOURNAL OF CHROMATOGRAPHY A 

2010;1217(4):565-574. 

797. Zeigler C, MacNamara K, Wang ZD, Robbat A. Total alkylated polycyclic 

aromatic hydrocarbon characterization and quantitative comparison of 

selected ion monitoring versus full scan gas chromatography/mass 

spectrometry based on spectral deconvolution. JOURNAL OF 

CHROMATOGRAPHY A 2008;1205(1-2):109-116. 

798. Zhang Y. Optimization of determination method for components and 

impurities of illicit drugs by GC-MS. ZHONGGUO YAOWU LANYONG 

FANGZHI ZAZHI 2007;13(1):19-21. 

7.4 High-Performance Liquid Chromatography (and tandem HPLC 

techniques): 

799. Dragovic S, Haghedooren E, Nemeth T, Palabiyik IM, Hoogmartens 

J, Adams E. Evaluation of two approaches to characterise liquid 

chromatographic columns using pharmaceutical separations. JOURNAL 

OF CHROMATOGRAPHY A 2009;1216(15):3210-3216. 

800. Dresen S, Gergov M, Politi L, Halter C, Weinmann W. ESI-MS/MS library 

of 1,253 compounds for application in forensic and clinical toxicology. 

ANALYTICAL & BIOANALYTICAL CHEMISTRY 2009;395(8):2521-2526. 

801. Fountain KJ, Xu J, Diehl DM, Morrison D. Influence of stationary phase 

chemistry and mobile-phase composition on retention, selectivity, and MS 

response in hydrophilic interaction chromatography. JOURNAL OF 

SEPARATION SCIENCE 2010;33(6-7):740-751. 

802. Gergov M. Forensic screening with liquid chromatography-mass 

spectrometry. HANDBOOK OF ANALYTICAL SEPARATIONS 

2008;6:491-511. 

803. Gholami M, Ghassempour A, Alizadeh R, Aboul-Enein HY. Microcolumn 

LC enantioseparation of chiral compounds using diol silica gel 

functionalized with vancomycin crystalline degradation products. 

JOURNAL OF SEPARATION SCIENCE 2009;32(7):918-922. 

804. Huerta-Fontela M, Galceran MT, Ventura F. Ultraperformance liquid 

chromatography-tandem mass spectrometry analysis of stimulatory drugs 

of abuse in wastewater and surface waters. ANALYTICAL CHEMISTRY 

2007;79(10):3821-3829. 

810

805. Huttunen J, Dawson M, Roux C, Robertson J. Physical evidence in drug 

intelligence Part 3: Supercritical fluid extraction-high performance liquid 

chromatography of packaging tapes. AUSTRALIAN JOURNAL OF 

FORENSIC SCIENCES 2009;41(1):63-72. 

806. Krawczeniuk AS. LC/MS Electrospray applications and the use of insource 

collision induced dissociation (CID) in the confirmatory analysis of 

drugs of abuse. 2008 American Academy of Forensic Sciences Annual 

Meeting. 

807. Lee H, Shen S, Grinberg N. Identification and control of impurities for drug 

substance development using LC/MS and GC/MS. JOURNAL OF LIQUID 

CHROMATOGRAPHY & RELATED TECHNOLOGIES 2008;31(15):2235- 

2252. 

808. Luo H, Ma LJ, Paek C, Carr PW. Application of silica-based hypercrosslinked 

sulfonate-modified reversed stationary phases for separating 

highly hydrophilic basic compounds. JOURNAL OF CHROMATOGRAPHY 

A 2008;1202(1):8-18. 

809. Lurie I, Li L. Use of high-temperature liquid chromatography with sub-2µm 

particle C18 columns for the analysis of seized drugs. JOURNAL OF 

LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES 

2009;32(18):2615-2626. 

810. Maldaner L, Jardim ICSF. A new thermally immobilized fluorinated 

stationary phase for RP-HPLC. JOURNAL OF SEPARATION SCIENCE 

2010;33(2):174-181. 

811. Min JZ, Hatanaka S, Toyo'oka T, Inagaki S, Kikura-Hanajiri R, Goda Y. 

Rapid sensitive and simultaneous determination of fluorescence-labeled 

designated substances controlled by the Pharmaceutical Affairs Law in 

Japan by ultra-performance liquid chromatography coupled with 

electrospray-ionization time-of-flight mass spectrometry. ANALYTICAL & 


812. Nakashima K, Ikeda R, Wada M. Analytical studies on the development of 

high-performance liquid chromatographic methods with fluorescence or 

chemiluminescence detections and their practical applications. 


813. Nguyen DT, Guillarme D, Heinisch S, Barrioulet MP, Rocca JL, Rudaz S, 

Veuthey JL. High throughput liquid chromatography with sub-2µm 

particles at high pressure and high temperature. JOURNAL OF 

CHROMATOGRAPHY A 2007;1167(1):76. 

814. Ojanpera, I, Pelander A, Ojanpera S. Comprehensive toxicological and 

forensic drug screening by LC/TOF-MS. CHEMICAL ANALYSIS 

2009;173:173-195. 

815. Ortiz RS, Antunes MV, Linden R. Determination Of sildenafil citrate and 

tadalafil by ultra performance liquid chromatography with diode-array 

detection (Uplc-Dad). QUIMICA NOVA 2010;33(2):389-393. 

811

816. Postigo C, de Alda MJL, Viana M, Querol X, Alastuey A, Artinano 

B, Barcelo D. Determination of drugs of abuse in airborne particles by 

pressurized liquid extraction and liquid chromatography-electrospraytandem 

mass spectrometry. ANALYTICAL CHEMISTRY 

2009;81(11):4382-4388. 

817. Ranc V, Havlicek V, Bednar P, Lemr K. Nanoelectrospray versus 

electrospray in chiral analysis by the kinetic method. COLLECTION OF 

CZECHOSLOVAK CHEMICAL COMMUNICATIONS 2009;74(2):313-322. 

818. Rodriquez-Cruz SE. Forensic Applications of liquid chromatography mass 

spectrometry (LC-MS). 2008 American Academy of Forensic Sciences 


819. Rosal C, Betowski D, Romano J, Neukom J, Wesolowski D, Zintek L. The 

development and inter-laboratory verification of LC-MS libraries for 

organic chemicals of environmental concern. TALANTA 2009;79(3):810- 

817. 

820. Sauvage FL, Gaulier JM, Lachatre G, Marquet P. Pitfalls and prevention 

strategies for liquid chromatography-tandem mass spectrometry in the 

selected reaction-monitoring mode for drug analysis. CLINICAL 

CHEMISTRY 2008;54(9):1519-1527. 

821. van Nuijs ALN, Tarcomnicu I, Bervoets L, Blust R, Jorens PG, Neels 

H, Covaci A. Analysis of drugs of abuse in wastewater by hydrophilic 

interaction liquid chromatography-tandem mass spectrometry. 


822. Wang Y, Ong TT, Li LS, Tan TTY, Ng SC. Enantioseparation of a novel 

chemistry derived native beta-cyclodextrin chiral stationary phase for highperformance 

liquid chromatography. JOURNAL OF CHROMATOGRAPHY 

A 2009;1216(12):2388-2393. 

823. Xiang P, Shen M, Zhuo X. Matrix effects in liquid chromatographic-mass 

spectrometric analysis. FENXI CESHI XUEBAO 2009;28(6):753-756. 

824. Zhang Y, Liang C, Ye H, Wang G, Wang R, Ni C, Liu Y, Zhang R. Method 

for simultaneously and qualitatively screening 242 kinds of compounds by 

liquid chromatography-tandem mass spectrometry (LC-MS/MS). 

APPLICATION: CN 2010-201228;15 October 2008. 

825. Zhen X, Bo Q, Lin GX, Lai CF, Hai L. Desorption electrospray ionization. 

PROGRESS IN CHEMISTRY 2008;20(4):594-601. 

7.5 Inductively Coupled Plasma- Mass Spectrometry (ICP-MS, Also ICP- 

OES): 

826. Ammann AA. Inductively coupled plasma mass spectrometry (ICP MS): a 

versatile tool. JOURNAL OF MASS SPECTROMETRY 2007;42(4):419- 

427. 

827. Ash RD, McDonough WF. In situ laser ablation icp ms trace element and 

isotope fingerprinting of powdered pharmaceuticals. The Fourth Forensic 

Isotope Ratio Mass Spectrometry Conference: 2010. 

812

828. Campbell GP, Curran JM, Miskelly GM, Coulson S, Yaxley GM, Grunsky 

EC, Cox SC. Compositional data analysis for elemental data in forensic 

science. FORENSIC SCIENCE INTERNATIONAL 2009;188(1-3):81-90. 

829. Noguchi O, Oshima M, Motomizu S. Minimization of sample volume with 

air-segmented sample injection and the simultaneous determination of 

trace elements by ICP-MS. ANALYTICAL SCIENCES 2008;24(5):631- 

635. 

830. Noguchi O, Oshima M, Motomizu S. Enrichment with air-sandwiched 

method of on-line collection/concentration using chelating resin and 

simultaneous determination of trace elements by inductively coupled 

plasma atomic emission spectrometry. TALANTA 2009;78(4-5):1235- 

1239. 

831. Shuttleworth S. LA-ICP-MS: The Ideal Tool Trace Element Fingerprinting 

of Solid Forensic Materials. Mid-Atlantic Association of Forensic Scientists 


832. van der Peijl G, van Breukelen M, van Es A, Wiarda W. Reporting of 

forensic iRMS/ICPMS results in The Netherlands. The Fourth Forensic 


833. Yang XJ, Low GK. Validation of a digestion procedure for ICP-AES and 

dynamic reaction cell ICP-MS for trace elemental analysis in 

environmental samples. ENVIRONMENTAL CHEMISTRY LETTERS 

2009;7(4):381-387. 

7.6 Infrared and Near Infrared Spectroscopy (including terahertz 

spectroscopy): 

834. Adam C. Shedding light on evidence: Forensic applications of UV/visible 

spectroscopy. SPECTROSCOPY EUROPE 2009;21(2):13-16. 

835. Ali HR, Edwards HG, Scowen IJ. Noninvasive in situ identification and 

band assignments of some pharmaceutical excipients inside USP vials 

with FT-near-infrared spectroscopy. SPECTROCHIMICA ACTA, PART A: 

MOLECULAR AND BIOMOLECULAR SPECTROSCOPY 2009;72(4):890- 

896. 

836. Bradley M, Izzia F, Nunn S. Analysis of mixtures by FT-IR: Spatial and 

spectral separation of complex samples. SPECTROSCOPY 2008;20- 

22:24-26. 

837. Bukowski EJ, Monti JA. FTIR-ATR spectroscopy for identification of illicit 

drugs seized from clandestine laboratories. AMERICAN LABORATORY 

2007;39(20):16-17,19. 

838. Burnett AD, Fan WH, Upadhya PC, Cunningham JE, Hargreaves 

MD, Munshi T, Edwards HGM, Linfield EH, Davies AG. Broadband 

terahertz time-domain spectroscopy of drugs-of-abuse and the use of 

principal component analysis. ANALYST 2009;134(8):1658-1668. 

813

839. Clark D, Pysik A. The analysis of pharmaceutical substances and 

formulated products by vibrational spectroscopy. APPLICATIONS OF 

VIBRATIONAL SPECTROSCOPY IN PHARMACEUTICAL RESEARCH 

AND DEVELOPMENT 2007;213-238. 

840. Dolbnev, DV, Dorofeev VL, Arzamastsev SP, Azimova ID, Vakhtel AV, 

Stepanova EV. Use of near-infrared spectrophotometry (NIR) for 

identification of pharmaceutical drugs. VOPROSY BIOLOGICHESKOI, 

MEDITSINSKOI IFARMATSEVTICHESKOI KHIMII 2008;(6):27-30. 

841. El-Bardicy MC, Lotfy HM, El-Sayed MA, El-Tarras MF. Smart stabilityindicating 

spectrophotometric methods for determination of binary 

mixtures without prior separation. JOURNAL OF AOAC INTERNATIONAL 

2008;91(2):299-310. 

842. Galloway CM, Le Ru EC, Etchegoin PG. An iterative algorithm for 

background removal in spectroscopy by wavelet transforms. APPLIED 

SPECTROSCOPY 2009;63(12):1370-1376. 

843. Kazarian SG, Chan KLA. Micro- and Macro-Attenuated total reflection 

fourier transform infrared spectroscopic imaging. APPLIED 

SPECTROSCOPY 2010;64(5):135A-152A. 

844. Kocak A, Lucania JP, Berets SL. Some advances in fourier transform 

infrared transflection analysis and potential applications in forensic 

chemistry. APPLIED SPECTROSCOPY 2009;63(5):507-511. 

845. Lanzarotta A, Baumann L, Story GM, Witkowski MR, Khan F, Sommers A, 

Sommer AJ. Rapid molecular imaging using attenuated total internal 

reflection planar array infrared spectroscopy for the analysis of counterfeit 

pharmaceutical tablets. APPLIED SPECTROSCOPY 2009;63(9):979-991. 

846. Lei Y, Luo ZY, Hu CQ. Rapidly screening counterfeit drugs using near 

infrared spectroscopy: Combining qualitative analysis with quantitative 

analysis to increase effectiveness. JOURNAL OF NEAR INFRARED 

SPECTROSCOPY 2008;16(3):349-355. 

847. Li X, Liu B, Zhu K, Tu D. The application of near infrared spectral 

characteristic peaks in counterfeit drugs analysis. ZHONGGUO YAOSHI 

(BEIJING, CHINA) 2008;22(7):558-9. 

848. Lin ACY, Hsieh HM, Tsai LC, Linacre A, Lee JCI. Forensic applications of 

infrared imaging for the detection and recording of latent evidence. 


849. Liu GF, Zhao FW, Ge M, Wang WF. Application of terahertz time domain 

spectroscopy to explosive and illegal drug. SPECTROSCOPY AND 

SPECTRAL ANALYSIS 2008;28(5):966-969. 

850. Ng PH, Walker S, Tahtouh M, Reedy B. Detection of illicit substances in 

fingerprints by infrared spectral imaging. ANALYTICAL & 

BIOANALYTICAL CHEMISTRY 2009. 

851. Polli JE, Hoag SW, Flank S. Near-infrared spectrophotometric comparison 

of authentic and suspect pharmaceuticals. PHARMACEUTICAL 

TECHNOLOGY 2009;33(8):46-52. 

814

852. Puchert T, Lochmann D, Menezes JC, Reich G. Near-infrared chemical 

imaging (NIR-CI) for counterfeit drug identification-A four-stage concept 

with a novel approach of data processing (Linear Image Signature). 

JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSES 

2009;51(1):138-145. 

853. Roggo Y, Gendrin C, Spiegel C. Near infrared chemical imaging for the 

pharmaceutical industry. SPECTRA ANALYSE 2007;36(258):26-30. 

854. Rutlidge HT, Reedy BJ. Classification of heterogeneous solids using 

infrared hyperspectral imaging. APPLIED SPECTROSCOPY 

2009;63(2):172-179. 

855. Sadlej-Sosnowska N, Ocios A. Selectivity of identification of sugars using 

IR spectroscopy. CHEMIA ANALITYCZNA 2008;53(5):639-649. 

856. Sakamoto T, Fujimaki Y, Kawanishi T, Hiyama Y. An approach for 

qualitative analysis of pharmaceuticals using diffusion reflectance NIR 

spectroscopy. IYAKUHIN KENKYU 2009;40(7):387-401. 

857. Sasaki Y, Hayashi A, Otani C, Kawase K. Noninvasive mail inspection 

system with terahertz radiation. APPLIED SPECTROSCOPY 

2009;63(1):81-86. 

858. Sato C, Furube A, Katoh R, Nonaka H, Inoue H. Non-destructive and 

discriminating identification of illegal drugs by transient absorption 

spectroscopy in the visible and near-IR wavelength range. JAPANESE 

JOURNAL OF APPLIED PHYSICS 2008;47(11):8583-8589. 

859. Shi ZQ, Anderson CA. Application of monte carlo simulation-based photon 

migration for enhanced understanding of near-infrared (NIR) diffuse 

reflectance. Part I: Depth of Penetration in Pharmaceutical Materials. 

JOURNAL OF PHARMACEUTICAL SCIENCES 2010;99(5):2399-2412. 

860. Storme-Paris I, Clarot I, Esposito S, Chaumeil JC, Nicolas A, Brion F, 

Rieutord A, Chaminade P. Near InfraRed Spectroscopy homogeneity 

evaluation of complex powder blends in a small-scale pharmaceutical 

preformulation process, a real-life application. EUROPEAN JOURNAL OF 

PHARMACEUTICS AND BIOPHARMACEUTICS 2009;72(1):189-198. 

861. Storme-Paris I, Rebiere H, Matoga M, Civade C, Bonnet PA, Tissier 

MH, Chaminade P. Challenging near infrared spectroscopy discriminating 

ability for counterfeit pharmaceuticals detection. ANALYTICA CHIMICA 

ACTA 2010;658(2):163-174. 

862. Wang L, Li SB, Meng JS, Hu C. Identification of counterfeit drugs by direct 

near-infrared spectra comparison. ZHONGGUO YAOSHI 2008;22(8):668- 

671. 

863. West MJ, Went MJ. The spectroscopic detection of drugs of abuse on 

textile fibres after recovery with adhesive lifters. FORENSIC SCIENCE 


815

7.7 Ion Mobility Spectrometry: 

864. Geraghty E, Kekukh V, Bunker SN. Nitrovanillin calibration agent for trace 

analyses using ion mobility spectrometry. US Appl. 2007/6,246;31 

December 2007. 

865. Kanu AB, Hill HH. Identity confirmation of drugs and explosives in ion 

mobility spectrometry using a secondary drift gas. TALANTA 2007;73:692- 

699. 

866. Khayamian T, Jafari MT. Design for electrospray ionization - ion mobility 

spectrometry. ANALYTICAL CHEMISTRY 2007;79(8):3199. 

867. Sherma J, John D, Larkin FH. Ion Mobility Spectrometry (IMS) and 

IMS/MS. JOURNAL OF AOAC INTERNATIONAL 2009;92(5):163A-168A. 

7.8 Mass Spectrometry (including ambient pressure techniques and 

Isotope Ratio): 

868. Alimpiev S, Grechnikov A, Sunner J, Borodkov A, Karavanskii V, 

Simanovsky Y, Nikiforov S. Gas chromatography/surface-assisted laser 

desorption ionization mass spectrometry of amphetamine-like compounds. 


869. Andrade FJ, Shelley JT, Wetzel WC, Webb MR, Gamez G, Ray SJ, Hieftje 

GM. Atmospheric pressure chemical ionization source. 2. Desorptionionization 

for the direct analysis of solid compounds. ANALYTICAL 

CHEMISTRY 2008;80(8):2654-2663. 

870. Binette MJ, Lafontaine P, Vanier M, Ng LK. Characterization of Canadian 

cigarettes using multi-stable isotope analysis by gas chromatography- 

Isotope Ratio Mass Spectrometry. JOURNAL OF AGRICULTURAL AND 

FOOD CHEMISTRY 2009. 

871. Calderone G, Serra F, Lees M, Mosandl A, Reniero F, Guillou C, Moreno- 

Rojas JM. Inter-laboratory comparison of elemental analysis and gas 

chromatography/combustion/isotope ratio mass spectrometry. II. delta N- 

15 measurements of selected compounds for the development of an 

isotopic Grob test. RAPID COMMUNICATIONS IN MASS 

SPECTROMETRY 2009;23(7):963-970. 

872. Carter JF, Hill JC, Doyle S, Lock C. Results of four inter-laboratory 

comparisons provided by the Forensic Isotope Ratio Mass Spectrometry 

(FIRMS) network. SCIENCE & JUSTICE 2009;49(2):127-137. 

873. Chen HW, Zheng J, Zhang X, Luo MB, Wang ZC, Qiao XL. Surface 

desorption atmospheric pressure chemical ionization mass spectrometry 

for direct ambient sample analysis without toxic chemical contamination. 

JOURNAL OF MASS SPECTROMETRY 2007;42(8):1045-1056. 

874. Collin OL. Development of a novel tandem mass spectrometry technique 

for forensic and biological applications. Avail UMI Order NO. DA3292877 

2007;68(12):7978. 

816

875. Collins M, Cawley A, Salouros H, Heagney A, Swan H, Vujic S. Stable 

isotope ratio methodologies for illicit drug intelligence programs. The 

Fourth Forensic Isotope Ratio Mass Spectrometry Conference: 2010. 

876. Cotte-Rodriguez I, Mulligan CC, Cooks G. Non-proximate detection of 

small and large molecules by desorption electrospray ionization and 

desorption atmospheric pressure chemical ionization mass spectrometry: 

Instrumentation and applications in forensics, chemistry, and biology. 

ANALYTICAL CHEMISTRY 2007;79(18):7069-7077. 

877. Dove A. Mass spectrometry raises the bar. SCIENCE 

2010;328(5980):920-922. 

878. Doyle S. Special issue: Forensic application of isotope ratio mass 

spectrometry (IRMS). SCIENCE & JUSTICE 2009;49(2):61-61. 

879. Ehleringer JR, Cerling TE, West JB, Podlesak DW, Chesson LA, Bowen 

GJ. Spatial considerations of stable isotope analyses in environmental 

forensics. ISSUES IN ENVIRONMENTAL SCIENCE AND TECHNOLOGY 

2008;26:36-53. 

880. Galhena AS, Harris GA, Nyadong L, Murray KK, Fernandez FM. Small 

molecule ambient mass spectrometry imaging by infrared laser ablation 

metastable-induced chemical ionization. ANALYTICAL CHEMISTRY 

2010:82(6):2178-2181. 

881. Galimov E, Sevastyanov V, Kuznetsova O. Isotope ratio mass 

spectrometry for tracing the origin of forensic agents. The Fourth Forensic 


882. Grange AH. An Integrated Wipe Sample transport/autosampler to 

maximize for a direct analysis in real time (DART)/orthogonal acceleartion, 

time-of-flight mass spectrometer (oa-TOFMS). ENVIRONMENTAL 

FORENSICS 2008;9(2-3):137-143. 

883. Green FM, Salter TL, Stokes P, Gilmore IS, O'Connor G. Ambient mass 

spectrometry: advances and applications in forensics. SURFACE AND 

INTERFACE ANALYSIS 2010;42(5):347-357. 

884. Haapala M, Pol J, Saarela V, Arvola V, Kotiaho T, Ketola RA, Franssila S, 

Kauppila TJ, Kostiainen R. Desorption atmospheric pressure 

photoionization. ANALYTICAL CHEMISTRY 2007;79(20):7867-7872. 

885. Hoogewerff J. The use of strontium isotope ratios and biological profiling 

for forensic provenancing of soils. The Fourth Forensic Isotope Ratio 

Mass Spectrometry Conference: 2010. 

886. Ifa DR, Jackson AU, Paglia G, Cooks RG. Forensic applications of 

ambient ionization mass spectrometry. ANALYTICAL & BIOANALYTICAL 

CHEMISTRY 2009;394(8):1995-2008. 

887. Ifa DR, Wu CP, Ouyang Z, Cooks RG. Desorption electrospray ionization 

and other ambient ionization methods: Current progress and preview. 

ANALYST 2010;135(4):669-681. 

888. Ishii A, Kaneko R, Hirata Y, Hamajima M. Application of new mass 

spectrometric techniques to legal medicine. JOURNAL OF THE MASS 

SPECTROMETRY SOCIETY OF JAPAN 2008;56(3):131-138. 

817

889. Jurschik S, Tani A, Sulzer P, Haidacher S, Jordan A, Schottkowsky R, 

Hartungen E, Hanel G, Seehauser H, Mark L, Mark TD. Direct aqueous 

injection analysis of trace compounds in water with proton-transferreaction 

mass spectrometry (PTR-MS). INTERNATIONAL JOURNAL OF 

MASS SPECTROMETRY 2010;289(2-3):173-176. 

890. Kala M. Forensic sciences. MASS SPECTROMETRY 2009:309-319. 

891. Kauppila TJ, Arvola V, Haapala M, Pol J, Aalberg L, Saarela V, Franssila 

S, Kotiaho T, Kostiainen R. Direct analysis of illicit drugs by desorption 

atmospheric pressure photoionization. RAPID COMMUNICATIONS IN 


892. Kauppila TJ, Kostiainen R, Kotiaho T, Laakkonen UM, Luosujarvi L. 

Analysis of street market confiscated drugs by desorption atmospheric 

pressure photoionization and desorption electrospray ionization coupled 

with mass spectrometry. RAPID COMMUNICATIONS IN MASS 

SPECTROMETRY 2009;23(9):1401-1404. 

893. Kawamura M, Kikura-Hanajiri R, Goda Y. Simple and rapid screening for 

psychotropic natural products using direct analysis in real time (DART)- 

TOFMS. YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL 

SOCIETY OF JAPAN 2009;129(6):719-725. 

894. Kennedy C, Bowen GJ, Ehleringer JR. Characterizing temporal variations 

in the isotopic composition of human drinking water. The Fourth Forensic 


895. Kertesz V, Van Berkel GJ. Improved desorption electrospray ionization 

mass spectrometry performance using edge sampling and a rotational 

sample stage. RAPID COMMUNICATIONS IN MASS SPECTROMETRY 

2008;22(23):3846-3850. 

896. Kertesz V, Van Berkel GJ. Improved imaging resolution in desorption 

electrospray ionization mass spectrometry. RAPID COMMUNICATIONS 

IN MASS SPECTROMETRY 2008;22(17):2639-2644. 

897. Landwehr J, Coplen TB. Spatial and seasonal variability of stable isotopic 

compositions of tap waters throughout the United States. The Fourth 


898. Liu K, Ma B, Wang Y, Chen X, Zhong D. A new software method for 

accurate mass measurements of drugs on unit mass resolution mass 

spectrometer. YAOXUE XUEBAO 2007;42(10):1112-1114. 

899. Liu YY, Lin ZQ, Zhang SC, Yang CD, Zhang XR. Rapid screening of 

active ingredients in drugs by mass spectrometry with low-temperature 

plasma probe. ANALYTICAL & BIOANALYTICAL CHEMISTRY 

2009;395(3):591-599. 

900. Luosujarvi L, Laakkonen UM, Kostiainen R, Kotiaho T, Kauppila TJ. 

Analysis of street market confiscated drugs by desorption atmospheric 

pressure photoionization and desorption electrospray ionization coupled 

with mass spectrometry. RAPID COMMUNICATIONS IN MASS 

SPECTROMETRY 2009;23(9):1401-1404. 

818

901. Martin AN, Farquar GR, Jones AD, Frank M. The non-destructive 

identification of solid over-the-counter medications using single particle 

aerosol mass spectrometry. RAPID COMMUNICATIONS IN MASS 


902. Martin AN, Farquar GR, Steel PT, Fergenson DP, Gard EE. The Rapid 

non-destruction identification of drug tablets using single particle aerosol 

mass spectrometry. 2008 American Academy of Forensic Sciences 


903. Martin AN, Farquar GR, Steele PT, Jones AD, Frank M. Use of single 

particle aerosol mass spectrometry for the automated nondestructive 

identification of drugs in multicomponent samples. ANALYTICAL 

CHEMISTRY 2009;81(22):9336-9342. 

904. Miao ZX, Chen H. Direct Analysis of liquid samples by desorption 

electrospray ionization-mass spectrometry (DESI-MS). JOURNAL OF 

THE AMERICAN SOCIETY FOR MASS SPECTROMETRY 

2009;20(1):10-19. 

905. Mitsui T. Effective use of mass spectrometry in scientific criminal 

investigation. JOURNAL OF THE MASS SPECTROMETRY OF JAPAN 

2008;56(3):117-122. 

906. Muccio Zeland, Jackson GP. Isotope ratio mass spectrometry. ANALYST 

2009;134(2):213-222. 

907. Mylonas R, Mauron Y, Masselot A, Binz PA, Budin N, Fathi M, Viette V, 

Hochstrasser DF, Lisacek F. X-Rank: A robust algorithm for small 

molecule identification using tandem mass spectrometry. ANALYTICAL 

CHEMISTRY 2009;81(18):7604-7610. 

908. Nilles JM, Connell TR, Durst HD. Thermal separation to facilitate Direct 

Analysis in Real Time (DART) of mixtures. ANALYST 2010;135(5):883- 

886. 

909. Oberacher H, Pavlic M, Libiseller K, Schubert B, Sulyok M, Schuhmacher 

R, Csaszar E, Kofeler HC. On the inter-instrument and inter-laboratory 

transferability of a tandem mass spectral reference library: 1. Results of an 

Austrian multicenter study. JOURNAL OF MASS SPECTROMETRY 

2009;44(4):485-493. 

910. Petucci C, Diffendal J, Kaufman D, Mekonnen B, Terefenko G, 

Musselman B. Direct analysis in real time for reaction monitoring in drug 

discovery. ANALYTICAL CHEMISTRY 2007;79(13):5064-5070. 

911. Philip RP, Kuder T. The evolution of stable isotope applications in 

environmental forensics-past, present and future. The Fourth Forensic 


912. Rao RM, Parab AR, Aggarwal SK. A study on the determination of 

strontium isotopic ratio by thermal ionization mass spectrometry using 

single as well as multiple filament assembly. BARC NEWSLETTER 

2007;285:80-83. 

819

913. Ratcliffe LV, Rutten FJM, Barrett DA, Whitmore T, Seymour D, 

Greenwood C, Aranda-Gonzalvo Y, Robinson S, McCoustra M. Surface 

analysis under ambient conditions using plasma-assisted 

desorption/ionization mass spectrometry. ANALYTICAL CHEMISTRY 

2007;79(16):6094-6101. 

914. Rodriguez-Cruz SE, Carson KA. Anion identification via complexation with 

meso-octamethylcalix(4)pyrrole and detection using electrospray 

ionization mass spectrometry. JOURNAL OF FORENSIC SCIENCES 

2010;55(2):499-507. 

915. Rodriguez-Cruz SE. Rapid screening of seized drug exhibits using 

desorption electrospray ionization mass spectrometry (DESI-MS). 


916. Shen C, Li J, Wang H, Han H, Xu G, Zheng P, Wang Y, Li H, Chu Y. Realtime 

measurements of the drug precursor chemicals with proton transfer 

reaction mass spectrometry. HUNAN DAXUE XUEBAO, ZIRAN 

KEXUEBAN 2009;36(2):75-79. 

917. Soparawalla S, Salazar GA, Perry RH, Nicholas M, Cooks RG. 

Pharmaceutical cleaning validation using non-proximate large-area 

desorption electrospray ionization mass spectrometry. RAPID 

COMMUNICATIONS IN MASS SPECTROMETRY 2009;23(1):131-137. 

918. Steiner RR, Larson RL. Validation of the direct analysis in real time 

Source for use in forensic drug screening. JOURNAL OF FORENSIC 

SCIENCES 2009;54(3):617-622. 

919. Talaty N, Mulligan CC, Justes DR, Jackson AU, Noll RJ, Cooks RG. 

Fabric analysis by ambient mass spectrometry for explosives and drugs. 

ANALYST 2008;133(11):1532-1540. 

920. Thevis M, Schrader Y, Thomas A, Sigmund G, Geyer H, Schaenzer W. 

Analysis of confiscated black market drugs using chromatographic and 

mass spectrometric approaches. JOURNAL OF ANALYTICAL 

TOXICOLOGY 2008;32(3):232-240. 

921. Van der Veer G, Voerkelius S, Lorentz G, Heiss G, Hoogewerff JA. Spatial 

interpolation of the deuterium and oxygen-18 composition of global 

precipitation using temperature as ancillary variable. JOURNAL OF 

GEOCHEMICAL EXPLORATION 2009;101(2):175-184. 

922. Walker GS. Combining Light Isotope, Trace Element and isotope of 

metals in environmental and forensic investigations. The Fourth Forensic 


923. West BJ, Ehleringer JR, Kreuzer HM, Hurley JM, Cerling TE. 

Provenancing plant materials form isoscapes. The Fourth Forensic 


924. Weston DJ. Ambient ionization mass spectrometry: Current understanding 

of mechanistic theory; analytical performance and application areas. 

ANALYST 2010;135(4):661-668. 

820

925. Wiseman JM, Laughlin BC. Desorption electrospray ionization (DESI) 

mass spectrometry: A brief introduction and overview. CURRENT 

SEPARATIONS AND DRUG DEVELOPMENT 2007;22(1):11. 

7.9 Microchip Technology: 

926. Bell SC, Hanes RD. A microfluidic device for presumptive testing of 

controlled substances. JOURNAL OF FORENSIC SCIENCES 

2007;52(4):884-888. 

927. Klenkar G, Liedberg B. A microarray chip for label-free detection of 

narcotics. ANALYTICAL & BIOANALYTICAL CHEMISTRY 

2008;391(5):1679-1688. 

7.10 Nuclear Magnetic Resonance Spectroscopy: 

928. Beyer T, Schollmayer C, Holzgrabe U. The role of solvents in the signal 

separation for quantitative H-1 NMR spectroscopy. JOURNAL OF 


929. Charlton AJ, Robb P, Donarski JA, Godward J. Non-targeted detection of 

chemical contamination in carbonated soft drinks using NMR 

spectroscopy, variable selection and chemometrics. ANALYTICA 

CHIMICA ACTA 2008;618(2):196-203. 

930. Chen SN, Turner A, Jaki BU, Nikolic D, van Breemen RB, Friesen JB, 

Pauli GF. An experimental implementation of chemical subtraction. 

JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS 

2008;46(4):692-698. 

931. Claridge TDW, Davies SG, Polywka MEC, Roberts PM, Russell AJ, 

Savory ED, Smith AD. "Pure by NMR"? ORGANIC LETTERS 

2008;10(23):5433-5436. 

932. Dai B, Eads CD. Efficient removal of unwanted signals in NMR spectra 

using the filter diagonalization method. MAGNETIC RESONANCE IN 


933. Dalisay DS, Molinski TF. NMR Quantitation of natural products at the 

nanomole scale. JOURNAL OF NATURAL PRODUCTS 2009;72(4):739- 

744. 

934. de Brouwer H. Evaluation of algorithms for automated phase correction of 

NMR spectra. JOURNAL OF MAGNETIC RESONANCE 2009;201(2):230- 

238. 

935. Hays PA, Thompson RA. A processing method enabling the use of peak 

height for accurate and precise proton NMR quantitation. MAGNETIC 

RESONANCE IN CHEMISTRY 2009;47(10):819-824. 

936. Henderson TJ. Feasibility study for the rapid screening of target molecules 

using translational diffusion coefficients: Diffusion-ordered NMR 

spectroscopy of biological toxins. ANALYTICAL & BIOANALYTICAL 


821

937. Mo HP, Harwood JS, Raftery D. Receiver gain function: The actual NMR 

receiver gain. MAGNETIC RESONANCE IN CHEMISTRY 

2010;48(3):235-238. 

938. Mo HP, Raftery D. Solvent Signal as an NMR Concentration Reference. 


939. Ozdemir MS, De Deene Y, Fieremans E, Lemahieu I. Quantitative proton 

magnetic resonance spectroscopy without water suppression. JOURNAL 

OF INSTRUMENTATION 2009;4. 

940. Pantoja-Uceda D, Santoro J. Aliasing in reduced dimensionality NMR 

spectra: (3,2)D HNHA and (4,2)D HN(COCA)NH experiments as 

examples. JOURNAL OF BIOMOLECULAR NMR 2009;45(4):351-356. 

941. Pauli GF, Jaki BU, Lankin DC. A routine experimental protocol for qHNMR 

illustrated with taxol. JOURNAL OF NATURAL PRODUCTS 

2007;70(4):589-595. 

942. Pierens GK, Carroll AR, Davis RA, Palframan ME, Quinn RJ. 

Determination of analyte concentration using the residual solvent 

resonance in H-1 NMR spectroscopy. JOURNAL OF NATURAL 

PRODUCTS 2008;71(5):810-813. 

943. Rubtsov DV, Griffin JL. Time-domain Bayesian detection and estimation of 

noisy damped sinusoidal signals applied to NMR spectroscopy. JOURNAL 

OF MAGNETIC RESONANCE 2007;188:367-379. 

944. Silvestre V, Mboula VM, Jouitteau C, Akoka S, Robins RJ, Remaud GS. 

Isotopic C NMR spectrometry to assess counterfeiting of active 

pharmaceutical ingredients: Site-specific C content of aspirin and 

paracetamol. JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL 

ANALYSIS 2009;50(3):336-341. 

945. Son H, Hwang G, Ahn H, Park W, Lee C, Hong Y. Characterization of 

wines from grape varieties through multivariate statistical analysis of H-1 

NMR spectroscopic data. FOOD RESEARCH INTERNATIONAL 

2009;42(10):1483-1491. 

946. Stanek J, Kozminski W. Iterative algorithm of discrete Fourier transform 

for processing randomly sampled NMR data sets. JOURNAL OF 

BIOMOLECULAR NMR 2010;47(1):65-77. 

947. Stern AS, Donoho DL, Hoch JC. NMR data processing using iterative 

thresholding and minimum l(1)-norm reconstruction. JOURNAL OF 

MAGNETIC RESONANCE 2007;188:295-300. 

948. Trefi S, Gilard V, Balayssac S, Malet-Martino M, Martino R. The 

usefulness of 2D DOSY and 3D DOSY-COSY H NMR for mixture 

analysis: Application to genuine and fake formulations of sildenafil 

(Viagra). MAGNETIC RESONANCE IN CHEMISTRY 2009;47(S1):S163- 

S173. 

949. Trefi S, Routaboul C, Hamieh S, Gilard V, Malet-Martino M, Martino R. 

Analysis of illegally manufactured formulations of tadalafil (Cialis) by H 

NMR, 2D DOSY H NMR and Raman spectroscopy. JOURNAL OF 


822

950. Wang T, Shao K, Chu QY, Ren YF, Mu YM, Qu LJ, He J, Jin CW, Xia B. 

Automics: An integrated platform for NMR-based metabonomics spectral 

processing and data analysis. BMC BIOINFORMATICS 2009;10. 

951. Webster GK, Marsden I, Pommerening CA, Tyrakowski CM, Tobias B. 

Determination of relative response factors for chromatographic 

investigations using NMR spectrometry. JOURNAL OF 

PHARMACEUTICAL AND BIOMEDICAL ANALYSIS 2009;49(5):1261- 

1265. 

952. Webster GK, Marsden I, Pommerening CA, Tyrakowski CM. Validation of 

pharmaceutical potency determinations by quantitative nuclear magnetic 

resonance spectrometry. APPLIED SPECTROSCOPY 2010;64(5):537- 

542. 

7.11 Raman Spectroscopy: 

953. Ali EMA, Edwards HGM, Hargreaves MD, Scowen IJ. In-situ detection of 

drugs-of-abuse on clothing using confocal Raman microscopy. 

ANALYTICA CHIMICA ACTA 2008;615(1):63-72. 

954. Anonymous. Innovative use of Raman spectroscopy as a forensic tool. 

BIOANALYSIS 2009;1(7):1189. 

955. Bell SEJ, Sirimuthu NMS. Quantitative surface-enhanced Raman 

spectroscopy. CHEMICAL SOCIETY REVIEWS 2008;37(5):1012-1024. 

956. Buckley K, Goodship A, MacLeod NA, Parker AW, Matousek P. 

Technique for enhancing signal in conventional backscattering 

fluorescence and Raman spectroscopy of turbid media. ANALYTICAL 

CHEMISTRY 2008;80(15):6006-6009. 

957. Clelland BL. Forensic applications of Raman microspectroscopy, capillary 

electrophoresis, chromatography, and mass spectrometry for the analysis 

of textile fibers, dyes, illicit drugs, and anticoagulant rodenticides. (Thesis) 

DISSERTATION ABSTRACTS INTERNATIONAL, B 2007;67(9):5041. 

958. de Veij M, Deneckere A, Vandenabeele P, de Kaste D, Moens L. 

Detection of counterfeit Viagra with Raman spectroscopy. JOURNAL OF 


959. De Veij M, Vandenabeele P, Moens L. Detection of counterfeit erectile 

dysfunction drugs with Raman spectroscopy. SPECTROSCOPY EUROPE 

2008;20(3):7-10. 

960. De Veij M, Vandenabelle P, DeBeer T, Remon JP, Moens L. Reference 

database of Raman spectra of pharmaceutical excipients. JOURNAL OF 

RAMAN SPECTROSCOPY 2009;40(3):297-307. 

961. Efremov EV, Ariese F, Gooijer C. Achievements in resonance Raman 

spectroscopy - review of a technique with a distinct analytical chemistry 

potential. ANALYTICA CHIMICA ACTA 2008;606(2):119-134. 

823

962. Hargreaves MD, Burnett AD, Munshi T, Cunningham JE, Linfield EH, 

Davies AG, Edwards HGM. Comparison of near infrared laser excitation 

wavelengths and its influence on the interrogation of seized drugs-ofabuse 

by Raman spectroscopy. JOURNAL OF RAMAN 

SPECTROSCOPY 2009;40(12):1974-1983. 

963. Hargreaves MD, Page K, Munshi T, Tomsett R, Lynch G, Edwards HGM. 

Analysis of seized drugs using portable Raman spectroscopy in an airport 

environment - a proof of principle study. JOURNAL OF RAMAN 

SPECTROSCOPY 2008;39(7):873-880. 

964. Kiefer W. Recent advances in linear and non-linear Raman spectroscopy. 

Part III. JOURNAL OF RAMAN SPECTROSCOPY 2009;40(12):1766- 

1779. 

965. MacLeod NA, Matousek P. Emerging non-invasive Raman methods in 

process control and forensic applications. PHARMACEUTICAL 

RESEARCH 2008;25(10):2205-2215. 

966. McGoverin CM, Rades T, Gordon KC. Recent pharmaceutical applications 

of raman and terahertz spectroscopies. JOURNAL OF 

PHARMACEUTICAL SCIENCES 2008;97(11):4598-4621. 

967. Noonan KY, Tonge LA, Fenton OS, Damiano DB, Frederick KA. Rapid 

classification of simulated street drug mixtures using raman spectroscopy 

and principal component analysis. APPLIED SPECTROSCOPY 

2009;63(7):742-747. 

968. Otieno-Alego V. Some forensic applications of a combined micro-Raman 

and scanning electron microscopy system. JOURNAL OF RAMAN 

SPECTROSCOPY 2009;40(8):948-953. 

969. Roggo Y, Degardin K, Margot P. Identification of pharmaceutical tablets by 

Raman spectroscopy and chemometrics. TALANTA 2010;81(3):988-995. 

970. Virtanen S, Antikainen O, Yliruusi J. Determination of the crushing 

strength of intact tablets using Raman spectroscopy. INTERNATIONAL 

JOURNAL OF PHARMACEUTICS 2008;360(1-2):40-46. 

7.12 Solid Phase Micro-Extraction (Headspace Techniques and Solvent 

Analysis): 

971. Andrasko J. Some examples of applications of a microthermal desorption 

device in the forensic laboratory. JOURNAL OF FORENSIC SCIENCES 

2009;54(5):1055-1058. 

972. Beam KN, Brettell TA. Forensic applications of headspace single-drop 

microextraction. 2008 American Academy of Forensic Sciences Annual 

Meeting. 

973. Baliyan PK, Singh RP, Arora S. Simultaneous estimation of residual 

solvents (Ethanol, Acetone, Dichloromethane and Ethyl Acetate) in 

dosage form by GC-HS-FID. ASIAN JOURNAL OF CHEMISTRY 

2009;21(8):5851-5857. 

824

974. Kang XJ, Chen LQ, Zhang YY, Liu YW, Gu ZZ. Performance of 

electrospun nanofibers for SPE of drugs from aqueous solutions. 

JOURNAL OF SEPARATION SCIENCE 2008;31(18):3272-3278. 

975. Lai H, Guerra P, Joshi M, Almirall JR. Analysis of volatile components of 

drugs and explosives by solid phase microextraction-ion mobility 

spectrometry. JOURNAL OF SEPARATION SCIENCE 2008;31(2):402- 

412. 

976. Lai H, Corbin I, Almirall JR. Headspace sampling and detection of 

cocaine, MDMA, and marijuana via volatile markers in the presence of 

potential interferences by solid phase microextraction-ion mobility 

spectrometry (SPME-IMS). ANALYTICAL AND BIOANALYTICAL 

CHEMISTRY 2008;392(1-2):105-113. 

977. Lai HT, Almirall JR. Headspace Sampling and detection of cocaine, 

MDMA, and marijuana via volatile chemical markers; solid phase 

microextraction-ion mobility spectrometry. 2008 American Academy of 


978. Misharina TA. Determination of volatile organics in gaseous phase using 

porous adsorbents. JOURNAL OF ANALYTICAL CHEMISTRY 

2010;65(2):127-134. 

979. Ouyang G, Pawliszyn J. A critical review in calibration methods for solidphase 

microextraction. ANALYTICA CHIMICA ACTA 2008;627(2):184- 

197. 

980. Snow NH, Bullock GP. Novel techniques for enhancing sensitivity in static 

headspace extraction-gas chromatography. JOURNAL OF 

CHROMATOGRAPHY A 2010;1217(16):2726-2735. 

981. Wang YW, McCaffrey J, Norwood DL. Recent advances in headspace gas 

chromatography. JOURNAL OF LIQUID CHROMATOGRAPHY & 

RELATED TECHNOLOGIES 2008;31(11-12):1823-1851. 

7.13 Thin Layer Chromatography: 

982. Gheorghe M, Balalau D, Ilie M, Baconi DL, Ciobanu AM. Qualitative 

analysis of confiscated illegal drugs by thin-layer chromatography. 

FARMACIA 2008;56(5):541-546. 

983. Sherma J. Analysis of counterfeit drugs by thin layer chromatography. 

ACTA CHROMATOGRAPHICA 2007;19:5-20. 

984. Turkmen Z, Mercan S, Tutanc L. Simultaneous High Performance Thin 

Layer Chromatographic Determination of Heroin, Morphine, Cocaine, and 

MDMA. 2008 American Academy of Forensic Sciences Annual Meeting. 

825

7.14 X-Ray based Techniques: 

985. Causin V, Marega C, Carresi P, Schiavone S, Marigo A. A quantitative 

differentiation method for plastic bags by wide angle X-ray diffraction for 

tracing the source of illegal drugs. FORENSIC SCIENCE 


986. Cook E, Fong R, Horrocks J, Wilkinson D, Speller R. Energy dispersive Xray 

diffraction as a means to identify illicit materials: A preliminary 

optimization study. APPLIED RADIATION AND ISOTOPES 

2007;65(8):959. 

987. Kotrly M. Using X-ray diffraction in forensic science. ZEITSCHRIFT FUR 

KRISTALLOGRAPHIE 2007;222(3-4):193-198. 

988. Moore MD, Cogdill RP, Wildfong PLD. Evaluation of chemometric 

algorithms in quantitative X-ray powder diffraction (XRPD) of intact multicomponent 

consolidated samples. JOURNAL OF PHARMACEUTICAL 

AND BIOMEDICAL ANALYSIS 2009;49(3):619-626. 

989. Moore MD, Steinbach AM, Buckner IS, Wildfong PLD. A structural 

investigation into the compaction behavior of pharmaceutical composites 

using powder x-ray diffraction and total scattering analysis. 

PHARMACEUTICAL RESEARCH 2009;26(11):2429-2437. 

990. West M, Ellis AT, Potts PJ, Streli C, Vanhoof C, Wegrzynek D, 

Wobrauschek P. Atomic spectrometry update. X-Ray fluorescence 

spectrometry. JOURNAL OF ANALYTICAL ATOMIC SPECTROMETRY 

2009;24(10):1289-1326. 


991. Channaa H, Surmann P. Voltammetric analysis of N-containing drugs 

using the hanging galinstan drop electrode (HGDE). PHARMAZIE 

2009;64(3):161-165. 

992. Domenech A, Navarro P, Aran VJ, Muro B, Montoya N, Garcia-Espana E. 

Selective electrochemical discrimination between dopamine and 

phenethylamine-derived psychotropic drugs using electrodes modified 

with an acyclic receptor containing two terminal 3-alkoxy-5-nitroindazole 

rings. ANALYST 2010;135(6):1449-1455. 

993. Ferreira FJO, Crispim VR, Silva AX. Detection of drugs and explosives 

using neutron computerized tomography and artificial intelligence 

techniques. RADIATION AND ISOTOPES 2010;68(6):1012-1017. 

994. Frisk T, Sandstroem N, Eng L, van der Wijngaart W, Maansson P, 

Stemme G. An integrated QCM-based narcotics sensing microsystem. 

LAB ON A CHIP 2008;8(10):1648-57. 

995. Gayton-Ely M, Shakleya DM, Bell SC. Application of a pyroprobe to 

simulate smoking and metabolic degradation of abused drugs through 

analytical pyrolysis. JOURNAL OF FORENSIC SCIENCES 

2007;52(2):473. 

826

996. Hodge TA. Method and apparatus for forensic screening and laboratory 

information management systems. U.S. Patent Application Publication US 

20090275038 A1, November 2009. 

997. Hua Q. Radiocarbon: A chronological tool for the recent past. 

QUATERNARY GEOCHRONOLOGY 2009;4(5):378-390. 

998. Izake EL. Chiral discrimination and enantioselective analysis of drugs: An 

overview. JOURNAL OF PHARMACEUTICAL SCIENCES 

2007;96(7):1659. 

999. Jestice AL. Method and apparatus for detecting and classifying explosives 

and controlled substances. U.S. Pat. Appl. Publ. US 2009. 

1000. Kibar O, Chachisvilis M, Tu E, Marsilje TH. Separation and manipulation 

of a chiral object by rotating field. U.S. Pat Appl. Publ. US 2008 274, 555. 

1001. Li H. Rapid detection of drugs and poisons in forensic samples. 

HIROSHIMA DAIGAKU IGAKU ZASSHI 2007;55(4-6):25-33. 

1002. Liang M, Shen J, Wang G. Identification of illicit drugs by using SOM 

neural networks. JOURNAL OF PHYSICS D: APPLIED PHYSICS 

2008;41(13):135306/1-135306/6. 

1003. Molnar P, Thorey P, Bansaghi G, Szekely E, Poppe L, Tomin A, Kemeny 

S, Fogassy E, Simandi B. Resolution of racemic trans-1,2-cyclohexanediol 

with tartaric acid. TETRAHEDRON-ASYMMETRY 2008;19(13):1587- 

1592. 

1004. Perot B, Carasco C, Valkovic V, Sudac D, Franulovic A. Detection of illicit 

drugs with the EURITRACK system. AIP CONFERENCE PROCEEDINGS 

2009;1099:565-569. 

1005. Reeve S, Allen S. Multicolor cavity ringdown based detection method and 

apparatus used for detecting gas phase molecules emanating from 

explosives, drugs, hazardous materials, or patient’s breath. U.S. Pat. Appl. 

Publ. US 20090180119 A1 16 Jul 2009. 

1006. Reviriego F, Navarro P, Garcia-Espana E, Albelda MT, Frias JC, 

Domenech A, Yunta MJR, Costa R, Orti E. Diazatetraester 1 H-Pyrazole 

crowns as fluorescent chemosensors for AMPH, METH, MDMA (Ecstasy), 

and dopamine. ORGANIC LETTERS 2008;10(22):5099-5102. 

1007. Silverio FO, Barbosa LCA, Pilo-Veloso D. Pyrolysis as an analytical 

technique. QUIMICA NOVA 2008;31(6):1543-1552. 

1008. Tondre C, Parant S, Lemiere P, Gerardin C. On the use of colloidenhanced 

ultrafiltration in view of enantiomeric enrichments and limiting 

conditions. COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND 

ENGINEERING ASPECTS 2008;317(1-3):431-437. 

8. Portable Detection and Analytical Instrumentation 

Issue: 

“Free Trade” agreements and the easing of formally restrictive national and 

international borders have resulted in dramatic increases in cargo 

827

transshipments and personal travel, thereby complicating drug inspection and 

interdiction efforts at POEs. Discovery and confirmational analysis of suspected 

drugs in cargo or on individuals is severely hampered by the lack of on-site 

detection and/or analytical equipment. 

Solution: 

Development of portable and highly sensitive detectors for drug detection and 

analyses allows law enforcement personnel and/or forensic chemists to perform 

screening type analyses on-site. In those cases where new methodologies have 

proven effective, case reports are generated for the forensic and enforcement 

communities. 

8.1 General 

1009. Amisar S. A reagent, a kit, and a method for detecting and identifying a 

wide range of illicit drugs. US 2007-2007/PV935762;30 August 2007. 

1010. Babichenko S, Ivkina T, Poryvkina L, Sominsky V. Method for on-site drug 

detection in illicit drug samples. PCT Int. Appl. WO 2008. 

1011. Chen X, Lian Y, Li A, Li J, Liu G, He C, Wu X, Wang H, Li H. An intelligent 

recognition program for in-situ detection of illicit chemicals by ion mobility 

spectrometry. JISUANJI YU YINGYONG HUAXUE 2007;24(9):1145-1148. 

1012. Carron K, Cox R. Qualitative analysis and the answer box: a perspective 

on portable raman spectroscopy. ANALYTICAL CHEMISTRY 

2010;82(9):3419-3425. 

1013. Cleary M, Vitale M. Improvements to methods of in-field analysis. 

APPLICATION: WO 2009-GB225 26 Jan 2009. 

1014. Eliasson C, Macleod NA, Matousek P. Non-invasive detection of powders 

concealed within diffusely scattering plastic containers. VIBRATIONAL 

SPECTROSCOPY 2008;48(1):8-11. 

1015. Grates KM, Ring JG, Savage KA, Denicola TA. Conclusion of validation 

study of commercially available field test kits for common drugs of abuse. 


1016. Guerra-Diaz P, Gura S, Almirall JR. Dynamic planar solid phase 

microextraction-ion mobility spectrometry for rapid field air sampling and 

analysis of illicit drugs and explosives. ANALYTICAL CHEMISTRY 

2010;82(7):2826-2835. 

1017. O’Neil AJ, Jee RD, Lee G, Charvill A, Moffat AC. Use of a portable near 

infrared spectrometer for the authentication of tablets and the detection of 

counterfeit versions. JOURNAL OF NEAR INFRARED SPECTROSCOPY 

2008;16(3):327-333. 

1018. Quest DW, Horsley J. Field-test of a date-rape drug detection device. 

JOURNAL OF ANALYTICAL TOXICOLOGY 2007;31(6):354. 

828

1019. Ramsey SA, Mustacich RV, Smith PA, Hook GL, Eckenrode BA. Directly 

heated high surface area solid phase microextraction sampler for rapid 

field forensic analyses. ANALYTICAL CHEMISTRY 2009;81(21):8724- 

8733. 

1020. Staples EJ, Viswanathan S. Detection of contrabands in cargo containers 

using a high-speed gas chromatograph with surface acoustic wave 

sensor. INDUSTRIAL & ENGINEERING CHEMISTRY RESEARCH 

2008;47(21):8361-8367. 

1021. Wells JM, Roth MJ, Keil AD, Grossenbacher JW, Justes DR, Patterson 

GE, Barket DJ. Implementation of DART and DESI ionization on a 

fieldable mass spectrometer. JOURNAL OF THE AMERICAN SOCIETY 

FOR MASS SPECTROMETRY 2008;19(10):1419-1424. 

1022. Yoon SS. Portable counterfeit detector machine. PRIORITY: KR 2008- 

2293, 8 Jan 2008. 

1023. Zieba-Palus J, Borusiewicz R, Kunicki M. PRAXIS - combined m-Raman 

and m-XRF spectrometers in the examination of forensic samples. 

FORENSIC SCIENCE INTERNATIONAL 2008;175(1):1-10. 


9.1 Beverage Analysis: 

1024. Clark A. A device for automatically testing whether a drink has been 

spiked. APPLICATION: GB 2007-5716 24 March 2007. 

1025. Loane CJ. Chemical spot test for the detection of drugs of abuse in a 

beverage. (Patent) CHEMICAL ABSTRACTS 2007;147:501291d. 

1026. Madea B, Musshoff F. Knock-out drugs: Their prevalence, modes of 

action, and means of detection. DEUTSCHES ARZTEBLATT 

INTERNATIONAL 2009;106(20):341-U12. 

1027. Murray GM, Mason AF, Ott EW. Molecularly imprinted polymer and 

colorimetric indicator sensor device useful for testing presence of drugs or 

hormones in body fluids or beverages. U.S. Pat. Appl. Publ. US 

20090197297 A16 Aug 2009. 

1028. Petrenko ES, Petrenko AE, Petrenko DE. Mode and an arrangement for 

prevention of carrying in a luggage through an inspection zone of liquid 

explosive and narcotic substances in a household package for beverage 

and perfumery. (Patent) CHEMICAL ABSTRACTS 2007:1405564. 

9.2 Chemometrics: 

1029. Armstrong N, Hibbert DB. An introduction to Bayesian methods for 

analyzing chemistry data Part 1: An introduction to Bayesian theory and 

methods. CHEMOMETRICS AND INTELLIGENT LABORATORY 

SYSTEMS 2009;97(2):194-210. 

829

1030. Drabek J. Validation of software for calculating the likelihood ratio for 

parentage and kinship. FORENSIC SCIENCE INTERNATIONAL- 

GENETICS 2009;3(2):112-118. 

1031. Hibbert DB, Armstrong N. An introduction to Bayesian methods for 

analyzing chemistry data Part II: A review of applications of Bayesian 

methods in chemistry. CHEMOMETRICS AND INTELLIGENT 

LABORATORY SYSTEMS 2009;97(2):211-220. 

1032. Kafkafi N, Yekutieli D, Elmer GI. A data mining approach to in vivo 

classification of psychopharmacological drugs. 

NEUROPSYCHOPHARMACOLOGY 2009;34(3):607-623. 

1033. Liu Q, Zhang J, Chen L, Min F, He M. Intelligent analysis method for 

criminal case based on relevance. APPLICATION: CN 2010-50854 17 

December 2007. 

1034. Melucci D, Monti D, D’Elia M, Luciano G. Multivariate analysis of powder 

drugs using reflectance NIR spectrometry. CHIMICA E L’INDUSTRIA 

2009;91(4):83-89 

1035. Ratle F, Gagne C, Terrettaz-Zufferey AL, Kanevski M, Esseiva P, Ribaux 

O. Advanced clustering methods for mining chemical databases in 

forensic science. CHEMOMETRICS AND INTELLIGENT LABORATORY 

SYSTEMS 2008;90(2):123-131. 

1036. Zadora G. Glass analysis for forensic purposes - A comparison of 

classification methods. JOURNAL OF CHEMOMETRICS 2007;21(5- 

6):174-186. 

1037. Zadora G, Neocleous T, Aitken C. A Two-level model for evidence 

evaluation in the presence of zeros. JOURNAL OF FORENSIC 

SCIENCES 2010;55(2):371-384. 

1038. Zadora G, Neocleous T. Likelihood ratio model for classification of forensic 

evidence. ANALYTICA CHIMICA ACTA 2009;642(1-2):266-278. 

1039. Zadora G, Wolanska-Nowak, P. Applications of Bayesian networks in 

forensic genetics and criminalistics. Z ZAGADNIEN NAUK SADOWYCH 

2009;78:141-152. 

1040. Zadora G. Evaluation of evidence value of glass fragments by likelihood 

ratio and Bayesian Network approaches. ANALYTICA CHIMICA ACTA 

2009;642(1-2):279-290. 

9.3 Counterfeit Drugs: 

1041. Balogh MP. Techniques for structure elucidation of unknowns: Finding 

substitute active pharmaceutical ingredients in counterfeit medicine. 

LCGC NORTH AMERICA 2007;25(6):554. 

1042. Balogh MP. Techniques for structure elucidation of unknowns: Finding 

substitute active pharmaceutical ingredients in counterfeit medicines. LC- 

GC EUROPE 2008;21(2):84-95. 

1043. Dubois J, Wolff J-C, Warrack JK, Schoppelrei J, Lewis EN. NIR chemical 

imaging for counterfeit pharmaceutical products analysis. 

SPECTROSCOPY 2007;22(Suppl.):36. 

830

1044. Fernandez FM, Green MD, Newton PN. Prevalence and detection of 

counterfeit pharmaceuticals: A mini review. INDUSTRIAL & 

ENGINEERING CHEMISTRY RESEARCH 2008;47(3):585-590. 

1045. Fernandez FM, Hampton CY, Nyadong L, Navare A, Kwasnik M. Liquid 

chromatography and ambient ionization time-of-flight mass spectrometry 

for the analysis of genuine and counterfeit pharmaceuticals. CHEMICAL 

ANALYSIS 2009;173:113-132. 

1046. Francis AR, Lyle RD, Moses VL, Pichardo D. Authentication of 

pharmaceuticals using molecular computational identification. U.S. Pat. 

Appl. Publ. US 20090187364 A1 23 Jul 2009. 

1047. Kogan, V, Beckers D, Bolze J, Lehmann C, Schweim H, Steffens, K. 

Detection of counterfeit drugs in packaged products, for example in blister 

packs, by angle dispersive x-ray diffraction. Europe Patent Application, EP 

2,090,883, August 2009 

1048. Nyadong L, Harris GA, Balayssac S, Galhena AS, Malet-Martino M, 

Martino R, Parry RM, Wang MDM, Fernandez FM, Gilard V. Combining 

two-dimensional diffusion-ordered nuclear magnetic resonance 

spectroscopy, imaging desorption electrospray ionization mass 

spectrometry, and direct analysis in real-time mass spectrometry for the 

integral investigation of counterfeit pharmaceuticals. ANALYTICAL 

CHEMISTRY 2009;81(12):4803-4812. 

1049. Soltaninejad K, Faryadi M, Akhgari M, Bahmanabadi L. Chemical profile of 

counterfeited buprenorphine vials seized in Tehran, Iran. FORENSIC 

SCIENCE INTERNATIONAL 2007;172(2-3):e4. 

1050. Willis RC. Noninvasive testing for counterfeit drugs. ANALYTICAL 

CHEMISTRY 2007;79(5):1773. 

1051. Yi X, Yang Y. Method for identifying genuine traditional Chinese medicine 

based on stable isotope fingerprint. APPLICATION: CN 2010-55059;30 

August 2007:9. 

9.4 Dragon’s Blood: 

1052. Gupta D, Bleakley B, Gupta RK. Dragon's blood: Botany, chemistry and 

therapeutic uses. JOURNAL OF ETHNOPHARMACOLOGY 

2008;115(3):361-380. 

1053. Wu Z, Shu T, Zhang L, Zhang Q, Zhu H. Determination of cochinchinenin 

C new compound of Dragon’s blood with RP-HPLC. WUJING 

YIXUEYUAN XUEBAO 2008;17(5):417-419. 

9.5 Drugs on Currency: 

1054. Armeta S, de la Guardia M. Analytical methods to determine cocaine 

contamination of banknotes from around the world. TrAC-TRENDS IN 


831

1055. Di Donati E, Martin CCS, Spinosa De Martinis B. Determination of cocaine 

in Brazilian paper currency by capillary gas chromatography/mass 

spectrometry. QUIMICA NOVA 2007;30(8):1966-1967. 

1056. Ebejer KA, Lloyd GR, Brereton RG, Carter JF, Sleeman R. Factors 

influencing the contamination of UK banknotes with drugs of abuse. 


1057. Zuo YG, Zhang K, Wu JP, Rego C, Fritz J. An accurate and 

nondestructive GC method for determination of cocaine on US paper 

currency. JOURNAL OF SEPARATION SCIENCE 2008;31(13):2444- 

2450. 

9.6 Education: 

1058. Anonymous. Forensic science badly lacking in the United States. Nature 

2009;457(733):1071. 

1059. Boucher MA, Specht KM. A forensic-themed case study for the organic 

lab. JOURNAL OF CHEMICAL EDUCATION 2009;86(7):847-848. 

1060. Bowen A. Putting chemistry in context: The role of the light microscope in 

non-routine analysis. MICROSCOP 2007;55(4):147-161. 

1061. Coleman WF. Molecular models of real and mock illicit drugs from a 

forensic chemistry activity. JOURNAL OF CHEMICAL EDUCATION 

2008;85(6):880. 

1062. Harmon KJ, Miller LM, Millard JT. Crime scene investigation in the art 

world: the case of the missing masterpiece. JOURNAL OF CHEMICAL 

EDUCATION 2009;86(7):817-819. 

1063. Harpp DN. Crime in the classroom Part IV: Conclusions. JOURNAL OF 

CHEMICAL EDUCATION 2008;85(6):805-806. 

1064. Hasan S, Bromfield-Lee D, Oliver-Hoyo MT, Cintron-Maldonado JA. Using 

laboratory chemicals to imitate illicit drugs in a forensic chemistry activity. 

JOURNAL OF CHEMICAL EDUCATION 2008;85(6):813-816. 

9.7 Legal Issues: 

1065. Bono JP, Siegel JA. Pattern evidence and conformance to the 

requirements of Daubert. 2008 American Academy of Forensic Sciences 


1066. Bower CM. Forensics Under fire: are bad science and dueling experts 

corrupting criminal justice? SCIENCE 2008;320(5884):1722. 

1067. Malkoc E, Neuteboom W. The current status of forensic science laboratory 

accreditation in Europe. FORENSIC SCIENCE INTERNATIONAL 

2007;167(2-3):121-126. 

1068. Melton L. Courtroom chemistry. CHEMISTRY WORLD 2007;4(11):58-61. 

1069. Moskvina TP, Usov AI Kuz'min SA. Laboratories for forensic expertise and 

their accreditation problems. INORGANIC MATERIALS 2008;44(14):1612- 

1616. 

832

1070. Wadell-Smith R, McGuffin VL. The need for research in forensic science. 


9.8 Quality Assurance: 

1071. Bogusz MJ. Quality assurance aspects of identification with 

chromatographic-mass spectrometric methods. Z ZAGADNIEN NAUK 

SADOWYCH 2009;77:7-19. 

1072. Gates K, Chang N, Dilek I, Jian H, Pogue S, Sreenivasan U. The 

uncertainty of reference standards-a guide to understanding factors 

impacting uncertainty, uncertainty calculations, and vendor certifications. 

JOURNAL OF ANALYTICAL TOXICOLOGY 2009;33(8):532-539. 

1073. Hauck WW, Abernethy DR, Williams RL. Metrologic approaches to setting 

acceptance criteria: Unacceptable and unusual characteristics. JOURNAL 

OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS 2008;48(3):1042- 

1045. 

1074. Neumann C, Margo P. New perspectives in the use of ink evidence in 

forensic science: Part I. Development of a quality assurance process for 

forensic ink analysis by HPTLC. FORENSIC SCIENCE INTERNATIONAL 

2009;185(1-3):29-37. 

1075. Neumann C, Margot P. New perspectives in the use of ink evidence in 

forensic science Part II. Development and testing of mathematical 

algorithms for the automatic comparison of ink samples analysed by 

HPTLC. FORENSIC SCIENCE INTERNATIONAL 2009;185(1-3):38-50. 

1076. Peters FT, Drummer OH, Musshoff F. Validation of new methods. 


1077. Willink R. Uncertainty in repeated measurement of a small non-negative 

quantity: Explanation and discussion of Bayesian methodology. 

ACCREDITATION AND QUALITY ASSURANCE 2010;15(3):181-188. 

9.9 Sampling Plans: 

1078. Alberink I, Bolck A, Stoel RD. Comparison of frequentist methods for 

estimating the total weight of consignments of drugs. JOURNAL OF 


1079. Levy R, Zelkowicz A. Efficiency evaluation of a police operation to fight the 

drug plague: Distribution unit weight as an objective index. JOURNAL OF 

FORENSIC SCIENCES 2007;52(4):909. 

1080. Majid MA. Techniques in drug sampling. 2008 American Academy of 


1081. Mario JR. A probability-based sampling approach for the analysis of drug 

seizures composed of multiple containers of either cocaine, heroin, or 

cannabis. FORENSIC SCIENCE INTERNATIONAL 2010;197(1-3):105- 

113. 

833

9.10 Toolmarks: 

1082. Buckleton J, Triggs C, Taroni F, Champod C, Wevers G. Experimental 

design for acquiring relevant data to address the issue of comparing 

consecutively manufactured tools and firearms. SCIENCE & JUSTICE 

2008;48(4):178-181. 

1083. Nichols RG. Defending the scientific foundations of the firearms and tool 

mark identification discipline: Responding to recent challenges. JOURNAL 

OF FORENSIC SCIENCES 2007;52(3):586-594. 

1084. Nichols RG. Defending the scientific foundations of the firearms and tool 

mark identification discipline: responding to recent challenges - Response. 


9.11 Other: 

1085. Lynch JD, Arroyo SB. Risks to Colombian Amphibian Fauna from 

Cultivation of Coca (Erythroxylum coca): A Geographical Analysis. 

JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH-PART 

A-CURRENT ISSUES 2009;72(15-16):974-985. 

1086. Prevatt-Smith KM, Prisinzano TE. New therapeutic potential for 

psychoactive natural products. NATURAL PRODUCT REPORTS 

2010;27(1):23-31. 

1087. Scherer UW, Jacobi M, Castillo J, Forstel DH. Ultra-low-level 

measurements of 3H and 14C in wines and champagne. RADIATION 

EFFECTS AND DEFECTS IN SOLIDS 2009;164(5-6):382-385. 

1088. Virtanen S, Antikainen O, Raikkonen H, Yliruusi J. Granule Size 

Distribution of Tablets. JOURNAL OF PHARMACEUTICAL SCIENCES 

2010;99(4):2061-2069. 

1089. Wang N, Shen C, ding P, Yi W, sun W, Liu K, Ding, X, Fu D, Yuan J. The 

application of bomb C in forensics: Determination of date of birth for 

unknown victims. DIQIU HUAXUE 2009;38(4):387-392. 

1090. Wassenaar LI, Van Wilgenburg SL, Larson K, Hobson KA. A groundwater 

isoscape for Mexico. JOURNAL OF GEOCHEMICAL EXPLORATION 

2009;102(3):123-136. 

1091. Wu JJ. In situ test for determining whether items of real or personal 

property have been exposed to the manuf. of illegal drugs. APPLICATION: 

US2008-13558 14 January 2008. 

Notes: 

All categories are subdivided by topic or category, then alphabetically by the first 

author’s last name. 

following reference is law enforcement restricted and is not available to the 

general public: The Journal of the Clandestine Laboratory Investigating 

Chemists Association (all years) 

834

Examination of Firearms Review: 2007 to 2010 - Interpol

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