Techniques that acquire donor profiling information from fingermarks — A review

Techniques that acquire donor profiling information from fingermarks — A review

SCIJUS-00565; No of Pages 12 Science and Justice xxx (2015) xxx–xxx Contents lists available at ScienceDirect Science and Justice journal homepage: ...

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SCIJUS-00565; No of Pages 12 Science and Justice xxx (2015) xxx–xxx

Contents lists available at ScienceDirect

Science and Justice journal homepage: www.elsevier.com/locate/scijus

Review

Techniques that acquire donor profiling information from fingermarks — A review Annemieke van Dam a,⁎, Fleur T. van Beek a,1, Maurice C.G. Aalders a,b,1, Ton G. van Leeuwen a,1, Saskia A.G. Lambrechts a,1 a b

Department of Biomedical Engineering and Physics, University of Amsterdam, Academic Medical Center, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands Amsterdam Center for Forensic Science and Medicine (CLHC), University of Amsterdam, 1098 XH Amsterdam, The Netherlands

a r t i c l e

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Article history: Received 3 July 2015 Received in revised form 30 November 2015 Accepted 12 December 2015 Available online xxxx Keywords: Forensic Fingermarks Fingerprints Donor profiling Intelligence Review

a b s t r a c t Fingermarks are among the most important types of evidence that can be encountered at the scene of a crime since the unique ridge pattern of a fingerprint can be used for individualization. But fingermarks contain more than the characteristic pattern of ridges and furrows, they are composed of a wide variety of different components that originate from endogenous and exogenous sources. The chemical composition can be used to obtain additional information from the donor of the fingermark, which in turn can be used to create a donor profile. Donor profiling can serve at least two purposes i) to enhance the evidential value of fingermarks and ii) to provide valuable tactical information during the crime scene investigation. Retrieving this additional information is not limited to fingermarks that have been used for individualization, but can also be applied on partial and/or distorted fingermarks. In this review we have summarized the types of information that can be obtained from fingermarks. Additionally, an overview is given of the techniques that are available addressing their unique characteristics and limitations. We expect that in the nearby future, donor profiling from contact traces, including fingermarks will be possible. © 2015 The Chartered Society of Forensic Sciences. Published by Elsevier Ireland Ltd. All rights reserved.

Contents 1. 2. 3.

Introduction . . . . . . . . . . . . . . . . . . . . . . . . Fingermarks and their composition . . . . . . . . . . . . . Donor profiling information from fingermarks . . . . . . . . 3.1. DNA . . . . . . . . . . . . . . . . . . . . . . . . 3.1.1. Legal considerations . . . . . . . . . . . . . 3.2. Gender . . . . . . . . . . . . . . . . . . . . . . . 3.3. Age of the donor . . . . . . . . . . . . . . . . . . 3.4. Diet . . . . . . . . . . . . . . . . . . . . . . . . 3.5. Blood group type . . . . . . . . . . . . . . . . . . 3.6. Personal habits . . . . . . . . . . . . . . . . . . . 3.7. Other foreign materials: explosive- and gunshot residues 3.8. Health information . . . . . . . . . . . . . . . . .

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Abbreviations: ATR, attentuated total reflectance; DART, direct analysis in real time; DESI, desorption electro spray ionization; DNA, deoxyribonucleic acid; ECL, electrochemiluminescence; EDDP, 2-ethylidene-1,5-methyl-3,3-diphenyl-1-pyrroline; EGF, epidermal growth factor; FTIR, Fourier transform-infrared spectroscopy; GC, gas chromatography; GLC, gas–liquid chromatography; HPLC, high-performance-liquid-chromatography; HSI, hyper spectral imaging; IR, infrared spectroscopy; LC, liquid chromatography; MALDI, matrix-assisted laser desorption/ionization; MS, mass spectrometry; MSI, mass spectrometry imaging; PCA, principal component analysis; PETN, pentaerythritol nitrate; PLS, partial least squares; PLS–DA, partial least squares–discriminant analysis; RDX, hexahydro-1,3,5-trinitro-1,3,5-triazine; RS, Raman spectroscopy; SALDI, surface-assisted laser desorption/ionization; SERS, surface enhanced Raman spectroscopy; SIMS, secondary ion mass spectrometry; ssDNA, single stranded DNA; THC, Δ9-TETRAHYDROCANNABINOl; TNT, 2,4,6-trinitrotoluene; TOF, time-of-flight. ⁎ Corresponding author. Tel.: +31 20 566 4390. E-mail addresses: [email protected], [email protected] (A. van Dam). 1 Tel.: +31 20 566 4390.

http://dx.doi.org/10.1016/j.scijus.2015.12.002 1355-0306/© 2015 The Chartered Society of Forensic Sciences. Published by Elsevier Ireland Ltd. All rights reserved.

Please cite this article as: A. van Dam, et al., Techniques that acquire donor profiling information from fingermarks — A review, Sci. Justice (2015), http://dx.doi.org/10.1016/j.scijus.2015.12.002

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4. Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

1. Introduction Fingermarks contain an enormous amount of undisclosed information on the donor of the mark. One source of information can be deoxyribonucleic acid (DNA), which can give us personal identity information. But despite its presence in fingermarks, to our knowledge, DNA profiles obtained from fingermarks are seldom successfully used in forensic case work and in court, because of the low amount present. Nevertheless developments on DNA collection and typing are ongoing to make identification possible [1]. A limitation that should be kept in mind is that the collection of DNA from fingermarks is destructive to the pattern. In some cases, fingermarks can be photographed without any interference of a development technique. However, in most cases the fingermarks are latent and need to be developed. Most of the standard fingermark detection techniques can be used in sequence with DNA profiling, therefore a fingermark can be photographed before being swabbed for analysis [2– 4]. However, after development, not all fingermarks can be used for the identification process, as they may be smudged or distorted, in which case it is preferred to collect the fingermarks for DNA analysis as the present DNA might still reveal the identity of the donor. Additionally, successful DNA collection and typing depends on many variables including the age of the mark, the carrier of the fingermark and the fingermark development technique used as reviewed by Kumar et al. [5]. Besides the individualization of a donor, also donor characteristics such as race, hair colour, eye colour and height can be determined from the DNA as well [6,7]. Besides DNA, excreted metabolites, proteins, peptides and also exogenous components contain information about the donor, such as gender, blood group type, age, diet, drug use and health [8–14]. Currently, the challenge is to reliably retrieve this information from fingermarks as it is a minimal sample of complex origin. Since the acquisition of donor profiling information from contact traces is not integrated in crime scene investigation, we would like to bring the use of chemical composition of a fingermark to the attention of the forensic field by summarizing the different types of intelligence that it can contain. By highlighting the most relevant literature on the topic we would like to discuss the current state of donor profiling.

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usage or handling of certain items, such as explosives, may help in the verification or falsification of testimonies. Also, when individualization of the fingermark is not possible, for instance in case of a distorted or a badly developed fingermark the chemical composition of the fingermark can still be used to obtain tactical donor profiling information or even evidence when it is accepted as such by a judge. Fingermarks are composed of material derived from sweat excreted via the pores, which are present on the ridges, but can also be contaminated with other material originating from touching different body parts and exogenous components, such as food, cosmetics and drugs [15,17]. An excellent review on the different components present in fingermarks has been published by Girod et al. [18]. In short, the major source contributing to the composition of fingermarks is sweat, which can originate from the eccrine, sebaceous and/or apocrine glands. Eccrine glands are present all over the body and in highest density on the soles of the feet and palms of the hands and are therefore the main contributor to the chemical components present in the fingermark. Inorganic compounds, including ammonia, sodium, phosphate, fluoride and chloride and organic compounds, such as proteins and lipids, are excreted via the eccrine glands [17]. Sebaceous glands are located in areas of the body containing hair follicles and are most abundant in the facial region and are not present on the hand palms, fingertips and soles of feet. These glands secrete sebum, an oily material. As human behavior involves touching the face and other skin areas containing sebaceous glands, sebum can be found in fingermarks. Sebum components include triglycerides, squalene, wax esters, cholesterol and free fatty acids. The apocrine glands are highly distributed in the armpits and genital region and are the least studied glands, since contamination of apocrine excretion products with sebum complicates the study of the excretion products of apocrine glands [17]. Sweat is not the only source that contributes to the chemical composition of fingermarks. Environmental contaminants and endogenous body material, such as saliva, can also affect the composition of fingermarks. From all these components originating from different sources, donor profiling information can be obtained that may aid in the forensic investigation. 3. Donor profiling information from fingermarks

2. Fingermarks and their composition The friction ridge skin present on the soles of our feet, palms of our hands and tips of our fingers and toes is composed of ridges and grooves. When touching a surface with the fingertip, a specific pattern is left behind and is called a fingermark [15]. Fingermarks can be used for individualization and thus identification purposes, based on the assumption that the friction ridge pattern is characteristic for each individual, including the ridge pattern of identical twins, and that the friction ridge skin does not change over time, except in case of injury that affects the deeper layers of skin [16]. In crime scene investigation, the pattern of fingerprints has been used as an identification tool since the late 1800s [16]. Currently, fingermarks found at crime scenes are used for identification, verification and/or for mark to mark comparison using the pattern that is left behind. However, fingermarks contain more information than the ridge pattern only. Chemical knowledge on the composition might be used to increase the evidential value and obtain more information about the donor. With increasing the evidential value we mean that donor profiling may be used to get an indication about the donor’s characteristics in case of unknown donor, such as the gender or age of the donor and/or to assess a (witness) testimony. When the donor of the fingermark is known, information like drug

Donor profiling information is defined here as information additional to the fingermark ridge pattern which can tell us something about the donor. Types of intelligence gained by donor profiling can include DNA, gender or age determination, diet, blood group, drug use, explosives handling or even health status. How these types of intelligence can be obtained and how they may contribute to a case will be discussed in the sections below. 3.1. DNA One of the constituents in fingermarks is DNA, which can be used for individualization, but also contains additional information about the donor of the fingermark. The amount of DNA is in most cases much too low to ensure reliable analysis and varies from no detectable DNA to hundreds of picograms [2,19–22]. Most of the standard fingermark detection techniques can be used in sequence with DNA profiling, therefore a fingermark can be captured before being swabbed for analysis [2– 4]. However, to our knowledge, DNA profiles obtained from fingermarks are seldom successfully used in forensic case work and in court [23–25]. Other morphological characteristics such as race, hair colour, eye colour and height can be determined from the DNA as well.

Please cite this article as: A. van Dam, et al., Techniques that acquire donor profiling information from fingermarks — A review, Sci. Justice (2015), http://dx.doi.org/10.1016/j.scijus.2015.12.002

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3.1.1. Legal considerations Some countries have legislation preventing them from obtaining donor characteristics from DNA. Therefore, care must be taken whether knowledge of donor profiling information is allowed in one’s jurisdiction. For example in the Netherlands the determination of sex, race and eye colour is only allowed when the donor is unknown, additional characteristics can be determined but only when authorized by special request [6,7]. In England there is no specific definition in the law addressing what can and cannot be used, therefore, in practice the morphological characteristics can be determined and are allowed as evidence. In the US, the legal status of DNA phenotyping largely depends on state law. In Germany and Belgium only the determination of sex is allowed and the determination of any other morphological features is forbidden [6,7]. The use of the chemical composition (besides DNA) of fingermarks has not been prohibited and can greatly enhance the evidential value of fingermarks as well as providing valuable tactical information, development of the methods extracting this information from fingermarks and their application can have great impact on casework. Whether or not donor profiling should be limited due to privacy infringement as is done for DNA in some countries is a valid discussion point. 3.2. Gender The fingermarks of men and women seem to differ in two ways: the ridges of men's fingermarks are coarser and their marks are greasier than those of women [26–29]. Several studies have investigated the use of the ridge density for gender determination, in which the ridge density was found to be higher in female fingermarks [26,28–31]. However, racial differences were observed in the mean value of ridge density within males and females, meaning that this method of ridge counting is not the gold standard for gender determination in multicultural populations [29]. Several studies have tried to determine the gender based on the chemical composition of the fingermark focussing on the fatty and oily components excreted via the sebaceous glands onto the skin [8,12,32– 36]. In the review from Girod et al. [18] gender determination has been shortly discussed. They found contradictory results between studies, which were caused by the focus on different chemical components, including lipids, chloride ions and urea [12,18,32,33,37]. Nazarro-Porro et al. [34] analysed the presence and abundance of various fatty acids obtained from the skin surface using gas–liquid chromatography (GLC) and observed differences in the amount and composition of the

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skin surface lipids varying with the age and gender of the donor. A remarkable observation was that Δ 9-type unsaturated fatty acids were found more often in samples obtained from female than male donors. However, skin swabs directly taken from different body parts were used and the amount of lipids in fingermarks was not specifically investigated. In another study, ten specific components were selected and analysed using gas chromatography–mass spectrometry (GC–MS), including myristic acid, pentadecanoic acid, palmitic acid, palmitoleic acid, methyl palmitate, methyl palmitoleate, methyl stearate, oleic acid, stearic acid and cholesterol. Their peak area ratios relative to squalene were calculated, as shown in Fig. 1 [32]. No significant differences could be observed between the average values of the components to squalene ratio and gender. However, the average values of palmitic acid, palmitoleic acid, and oleic acid were slightly higher in fingermarks from males. Croxton et al. [35] investigated the presence of fatty acids in fingermarks as well as the presence of amino acids. They found a different mean level of amino acids in female fingermarks, which appeared to be higher compared to the mean level of amino acids in male fingermarks. However, except for the presence of asparagine no significant differences were found in the amino acid levels. Also, a higher amount of fatty acids was found in male fingermarks, although the difference in level was not significant. A drawback of the research performed by Croxton et al. [35] was that the number of donors was limited to 9 male and 9 female donors. Hartzwell-Baguley et al. [12] noticed in an initial experiment a difference in urea levels between genders, however no statements were made on the precise levels of urea found, also no information was given on the sample-size of the study. Another study looked at triacylglycerols (TAGs) using laser desorption/ionization time-of-flight mass spectrometry (LDI-TOF MS), but could only detect the TAGs using LDI and could not find significant differences indicating gender [38]. In a study by Michalski et al. [36] GC–MS was used to discriminate between gender and race focussing on the presence of fatty acids and their methyl esters, however no discrimination could be made. Recently, matrix assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) in conjugation with the data analysis method partial least squares discriminant analysis (PLS–DA) was used by Ferguson et al. [8] to discriminate between genders, whereby they focussed on peptides and small proteins present in fingermark residues. Promising results were obtained leading to gender discrimination with 67.5% to 85% accuracy. Interestingly, three biomarkers were identified that can probably be used for gender classification, namely SSL-29, and LEK-45 for males and DCD-1L for females. In conclusion, more research should be performed on gender determination from fingermarks. A general observation is that in most studies a limited amount of fingermarks were investigated, which makes it hard to draw reliable conclusions. Most of the studies discussed in this section used GC–MS to investigate components of interest. This is a highly sensitive technique, which can detect components in nanogram levels and is able to identify and quantify components of interest. However, two major drawbacks of this technique are that i) it is limited to volatile components and ii) the technique is destructive as fingermarks need to be dissolved and vaporized for detection and analysis [35,39, 40]. In contrast, MALDI-MSI is minimally destructive and besides identification, characterization and quantification, this MS system can also be used for imaging. This means that not only chemical information can be obtained from the fingermark, but also information on the ridge pattern [41]. A combination of the morphological aspects of the fingermark and its chemical composition might result in the nearby future in gender determination [26–29]. 3.3. Age of the donor

Fig. 1. Comparison of female and male average percentages of different components and their area ratios relative to squalene plotted with ± one standard deviation of error. No significant difference could be observed between the average values of these components and gender. Reprinted, with permission from the Journal of Forensic Sciences, Volume 47, Issue 4, copyright ASTM International, 100 Barr Harbor Drive, West Conshohocken, PA 19083 [32].

Two aspects of the fingermark change when the donor ages: the ridge groove pattern becomes coarser and the onset of puberty clearly influences the composition of the fingermark [17,42].

Please cite this article as: A. van Dam, et al., Techniques that acquire donor profiling information from fingermarks — A review, Sci. Justice (2015), http://dx.doi.org/10.1016/j.scijus.2015.12.002

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A general observation made by different fingerprint examiners was that children's fingermarks are hard to find and to develop after 24 h, whereas fingermarks of adults can be developed much longer after deposition [33,43]. The main difference in chemical composition between childrens' fingermarks and adults' is that the marks from children contain higher concentrations of volatile unesterified free fatty acids, whereas in adults’ marks higher concentration of more stable components, like long-chained fatty acid esters were found [10,33,44,45]. This observation can be explained by the difference in activity of the sebum glands with increasing age. Ramasastry et al. [46] investigated whether differences could be found in skin surface lipids obtained from children at birth and at an age of 17. Low cholesterol levels were found at birth, and were variable up to the age of nine years, whereafter a decrease was observed. The inverse happened with wax esters with high levels at birth, followed by variable levels at the age from 0 to 4 years, but after the age of six an increase in the level was observed, which remained stable after the age of nine. In another study performed by Nazarro-Porro et al. [34] skin surface samples were investigated on the levels of Δ 9-type unsaturated fatty acids, including triglycerides, wax esters and sterols. A maximum level of Δ 9-type unsaturated fatty acids was found before puberty, then the level decreased, reaching a minimum at middle age (20–65 years) and rose again with senescence. However, these two studies were performed on samples obtained from different body parts and they did not investigate samples obtained from fingerprints or fingermarks. Hemmila et al. [10] designed a spectroscopic approach to estimate the age of the fingermark donor using Fourier transform-infrared spectroscopy (FTIR) on fingermark residues left by 78 individuals with ages varying from 4 to 68 years. Principal component analysis (PCA) was used to separate the sample set in four groups with ages of 4–5, 11– 14, 18-26 and 29–70 years old. The spectral region of 2800–300-cm−1 was found to be the most significant wavelength range to separate the sample set in the four groups. The absorption in this region is caused by the C–H stretching vibration frequencies of aliphatic group and is determined by the ratio between free long-chain fatty acids and their corresponding esters, which are known to change with age [10]. The results of Hemmila et al. [10] were supported by the results obtained by Antoine et al. [47] who were able to distinguish fingermarks from children from adults' fingermarks even when they were aged up to 4 weeks after deposition. In conclusion, there are currently no methods that are able to accurately estimate the age of the donor. The detection of volatile unesterified free fatty acids and long-chained fatty acids may result in a better estimation of the age of the donor. Also, other endogenous compounds including proteins and hormones are likely to provide additional information on the age of the donor, since their serum levels change with age [48,49]. However, the presence of hormones and their relation to age have not been studied in fingermark depositions, yet. A combination of different methods, for instance FTIR and a method that is able to detect hormones in fingermarks, may result in the estimation of the age of the donor. 3.4. Diet Eating behavior and diet of individuals is found to influence the body odor of humans [50,51]. Therefore, it is likely that specific components related to eating behavior can be found in sweat and hence in the fingermark residue. One study by Lambrechts et al. indicated food metabolites in fingermarks using thin layer chromatography (TLC) combined with fluorescence spectroscopy [9]. The goal of their study was to identify the fluorescent components responsible for the intrinsic fluorescence of fingermarks. Pheophorbide A and pheophytin, two metabolites of chlorophyll, a plant pigment, were indicated to be the source of two characteristic red fluorescent spots originating from fingermarks and

found on a developed TLC plate. Chlorophyll can be found in green vegetables and thus the presence of chlorophyll metabolites in fingermarks may provide information about the donor’s diet [9]. However, TLC combined with fluorescence spectroscopy is not able to identify specific components and the combination of these methods will only give an indication of possible components. Caffeine and its metabolites can be detected in fingermarks [11,52]. In a study performed by Kuwayama et al. [11] these components were detected before and after coffee intake in fingermarks and blood samples using liquid chromatography mass spectrometry (LC-MS). Caffeine and paraxanthine, the major metabolite of caffeine, could be detected in fingermark residues, but the levels of theobromine and theophylline were below the lower limit of quantification. In the fingermarks of all three subjects the amount of caffeine and paraxanthine was higher after coffee intake, presented in Fig. 2. Food metabolites and spices can be analysed with direct analysis in real time-mass spectrometry (DART-MS), suggesting that dietary information could also be obtained from fingermarks with this technique [53]. In conclusion, more research could be conducted to determine interesting food metabolites that could provide information on diet. Also, information on the uptake, metabolization and excretion rate of foodstuffs will be of additional value when dietary information is requested, since this could provide knowledge on the eating behavior of at least the last two days before deposition of the fingermark. The work presented by Kuwayama et al. [11] is a good first step in the determination of excretion rate of foodstuffs in fingermarks, however they only included a limited amount of volunteers. Nevertheless, the added value of dietary information for a forensic investigation might not be enormous, yet, one must consider that in diagnostic applications this information may have a larger impact, especially since obtaining fingermarks from a patient is a non-invasive sampling method. 3.5. Blood group type Immunoassays can be used to detect macromolecules in fingermarks [54–60]. One of the first research groups who described the use of an immunogenic technique to obtain donor profiling information from fingermarks was the group of Ishiyama et al. [59]. They were able to type the ABH-blood group type of fingermark donors using a mixed cell agglutination method. A total of 116 samples were typed of which only 11 were typed wrong or inconclusive. In preliminary work of our research group we investigate the detection of blood group antigens in fingermarks. We were also able to detect antigen A and B simultaneously in fingermark residues using immunolabelling (unpublished work). Blood group typing is thus possible in fingermarks, however the sensitivity and specificity of the immunolabelling method for blood group typing needs to be addressed with a large validation study. Alternatively, considering the immense developments in the sensitivity of DNA technology one may argue that further investment in donor profiling by blood group typing may be unwise as it may become redundant quickly. However, blood group typing can still be used as fast indicative screening method (within minutes) to include or exclude possible donors of a trace as a first guidance, whereas DNA analysis and typing will take much longer. 3.6. Personal habits Knowledge on the use of hair gels, soaps, hand lotions and cosmetics will give a better understanding of the lifestyle and personal hygiene habits of the donor and can help to reduce the possible pool of donors. Other interesting information that can be obtained from fingermarks includes smoking habits and drug usage. Different studies showed that cosmetic ingredients can be observed in contaminated fingermarks [12,61–63]. Successful identification of different cosmetics was possible in fingermarks contaminated with face

Please cite this article as: A. van Dam, et al., Techniques that acquire donor profiling information from fingermarks — A review, Sci. Justice (2015), http://dx.doi.org/10.1016/j.scijus.2015.12.002

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Fig. 2. Amounts of caffeine and paraxanthine found in fingermarks before and after coffee intake. Average values and standard deviations were calculated of four measurements. Number sign # represents the value under the lower limit of quantification (LOQ). Dashed line shows the values obtained from fingermarks before coffee intake (subject B and C) or the LOQ (subject A). * significant difference of p b 0.05 compared to values before coffee intake. This figure has been adapted and reprinted from Kuwayama K, Tsujikawa K, Miyaguchi H, Kanamori T, Iwata YT, Inoue H:, Time-course measurements of caffeine and its metabolites extracted from fingertips after coffee intake: a preliminary study for the detection of drugs from fingerprints. Analytical and bioanalytical chemistry 2012:1–8 [11]. Copyright 2012, with permission from Springer.

cream, body lotion, foundation and body butter using ATR-FTIR [61]. In fingermark residues from females analysed with GC–MS certain levels of hydrocarbons, including tetracosane and octacosane were found, which are likely to originate from cosmetics containing petroleum jelly, as well as levels of octylmethoxycinnamate a commonly used ingredient in UV-B sunscreen or cosmetic penetration enhancers [12]. Another contaminant, that is known to originate from lotions, hair products, body washes and tissues is the dimethyldioactadecylammonium ion, which was reported to be present in fingermarks and could be detected with MALDI-MSI [63]. A large variety of fatty acids, which are commonly found in fingermarks are also present in cosmetics, for instance squalene and cholesterol, which makes it hard to differentiate between endogenous fingermark components and exogenous cosmetic ingredients [62]. At this moment, it is not possible to distinguish specific cosmetic components in fingermarks based on their brand, making this type of information not directly of great forensic relevance. It is however, possible to detect and distinguish condom traces in fingermark residues as shown by Bradshaw et al. [104]. MALDI MSI, MS/MS, RS and ATR-FTIR have been used to analyse fingermarks contaminated with condom traces to detect condom lubricants. In some fingermarks, it was even possible to discriminate between condom residues originating from different condom brands/types. Information on the contamination with condom traces might be of additional value in the victim’s statement of sexual assault and can be used as additional intelligence in forensic cases [104]. Besides determining donor profiling information on the use of cosmetics and hygiene products, drug use can also be considered, for example by the application of immunogenic techniques. The use of immunogenic techniques on fingermarks has been described in a review by Wood et al. [64] and Hazarika and Russell [65]. Ishiyama et al. [60] continued their work [59], but instead of the determination of blood group type, they tried to identify the drug metabolites, morphine and methamphetamine in sweat using a homogenous enzyme immunoassay. Morphine and methamphetamine could both be detected in sweat, suggesting that these metabolites will also be present in fingermarks. Other

research groups have shown that a variety of drug metabolites can be detected in fingermarks using immunolabelling, including cotinine, methadone, benzoylecgonine and tetrahydrocannabinol. However in these studies proper control experiments were omitted [66–70]. Hazarika et al. [65,67–69] tried to combine the detection of specific drug metabolites with the visualization of the ridge details in fingermarks. Conjugation of antibodies to magnetic particles resulted in a clear developed fingermark. However, due to the lack of a positive control and the inclusion of more negative controls, which for example could have been performed by incubation of unconjugated magnetic particles to the fingermark, no reliable conclusion can be given about the specificity of this method. Besides detection of drug metabolites they also tried to discriminate smokers from non-smokers in fingermarks by detecting a metabolite of nicotine, cotinine, as marker for cigarette smoking. Nicotine is a component present in tobacco, which is metabolized in the human body to cotinine and other metabolites. Immunolabelling can be used to discriminate smokers from nonsmokers using specific antibodies to cotinine [66,68]. In another study using capillary scale ion chromatography for the discrimination of smokers versus non-smokers, it was found that the levels of thiocyanate and benzoate, metabolites related to smoking behavior, were more pronounced in fingermarks from smokers than non-smokers [69]. However, nicotine can also be found in different foods, such as potatoes, tomatoes and cauliflower [71], which means that the source of the metabolite cotinine may be disputed. We tried to reproduce the results presented by Hazarika et al., using their immunolabelling protocol, however, we were not able to obtain specific immunolabelling in fingermarks [56]. We found that the magnetic particles used in these studies bound non-specifically to fatty components in the fingermark residue when no blocking buffer was applied. A reproducible immunolabelling method has been developed by our group, which makes it possible to detect specific components present in the fingermark [55–58]. Fig. 3 illustrates the immunolabelling of a general peptide in fingermarks, dermcidin, left on a variety of plastic surfaces [58].

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Fig. 3. Immunolabelling of dermcidin on non-porous plastics using anti-dermcidin and a secondary antibody tagged with Cy3 [58]. A: Blue garbage bag. B: Black garbage bag. C: Ziploc bag. D: Plastic sheet. E: Sandwich bag. Scale bar is 0.5 cm. F: Magnified image of an immunolabelled fingermark left on a black garbage bag.

An interesting method was introduced by Xu et al. [72] who combined electrochemiluminescence (ECL) with immunolabelling. The detection of the antigen-antibody complex can be visualized using the electrochemical stimulation of light emission. Fingermarks were left on gold film electrodes and immunolabelling was performed on the fingermarks using the reaction between horse radish peroxidase and luminol as detection tag. Epidermal growth factor (EGF), lysozyme and dermcidin were detected in fingermarks. However, in the reported immunolabelling method no proper blocking steps and control experiments were described. Also, samples were dried before incubation of the secondary antibody and detection tags. Drying out of the sample between the different experimental steps needs to be avoided at all times, since it will cause non-specific antibody binding and background staining [72,73]. Song et al. presented immunolabelling in combination with surface enhanced Raman spectroscopy (SERS) to detect IgG in spiked fingermarks. The antibodies, anti-IgG, were conjugated to Ag nanoparticles, which made it possible to detect antibodies that targeted their antigens with SERS [74]. Recently, a competitive enzyme immunoassay to detect cocaine in fingermarks was introduced by van der Heide et al. In their work they describe a sensitive method that allows the quantitative detection of cocaine in samples obtained from general circulating banknotes and fingermarks obtained from volunteers known to be taking drugs [75]. Instead of antibodies, aptamers can also be used to detect macromolecules [76]. Cocaine metabolites could also be detected with aptamers conjugated to gold nanoparticles in fingermarks [64,77]. Other than immunogenic methods, infrared hyper spectral imaging (HSI) has also been used to detect contaminants in fingermark residues. Fingermarks, spiked with ibuprofen, vitamin C, non-dairy creamer and/ or sweetener were analysed using infrared HSI. A major drawback of this study is that spiked fingermarks were used, which probably gives no good reflection of a fingermarks deposited by a person who really handled the before mentioned items. Spectral imaging demonstrated that specific components in fingermark residue could be detected using automated recognitions. A spectral library was used that compared the intensity and position of the peaks [78,79]. Besides immunolabelling and HSI, other techniques are also available which are able to detect drug metabolites in fingermarks and sweat [14, 80–93]. Goucher et al. [94] detected benzodiazepine lorazepam and one of its metabolites, 3-O-glucuronide, in fingermarks using LC-MS/MS.

Lorazepam is a clinically known drug, administrated as treatment for anxiety. Volunteers were asked to administer a two mg tablet of lorazepam. Fingermarks were deposited at different time points after administration. The presence of lorazepam and 3-O-glucuronide could not be detected in single fingermarks, but only in 10 times stacked depositions. Two hours after administration the highest levels of 3-O-glucuronide and lorazepam were observed in the ten times loaded fingermarks. Twenty-four hours after intake 3-O-glucuronide could no longer be detected in fingermarks, whereas lorazepam could no longer be detected 8 hours after intake [94]. Terbafine, the active compound present in an anti-fungal drug, could be found in fingermarks deposited by a patient who took 1 tablet of drug per day over a course of 14 days. Fingermarks were collected at day 1, 3, 7 and 14 of administration. An increased level of terbafine could be detected in fingermarks up to 14 days using SALDI-MS [95]. Also, methadone and its metabolite 2-ethylidene-1,5-methyl-3,3-diphenyl-1pyrroline (EDDP) could be detected in fingermarks deposited by donors recruited from an NHS outpatients Methadone Maintanance Clinic [85]. Methadone was given in a daily dose to subjects to manage heroin dependence. In fingermarks deposited by control subjects no methadone and EDDP could be detected [85]. Kuwayama et al. [86] investigated the presence of analytes of a cold medicine in fingermarks, which contains ibuprofen, chlorpheniramine, dihydrocodeine and methylephedrine. Volunteers were asked to take the medicine and deposit fingermarks at different time points, specifically before intake, 2, 5 and 9 hours after intake and 1, 2, 3, 4 and 7 days after intake. The levels of the different analytes were measured using LC-MS-MS and compared with levels found in blood. All analytes could be detected in the fingermarks after administration of the drugs, except for ibuprofen. In blood samples ibuprofen could only be detected at 2 and 5 h after administration. In volunteers who did not administer the drugs, no analytes were detected in fingermarks or in the blood samples. Recently, it has been shown by Bailey et al. that MALDI-MS and DESI can be used to detect cocaine and its metabolites in fingermarks residues [91]. Fingermarks were obtained from five individuals who were attending a drug and alcohol treatment service. The results of the DESI and MALDI analysis of the fingermarks were compared with the outcome of oral fluids screening tests and a good correlation was found between the detection of the different metabolites, except for one fingermark in which the oral fluid screening test was able to detect cocaine, benzoylecgonine and methylecgonine, whereas DESI could only detect cocaine [91].

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In some cases the fingermarks used for detection of drug metabolites were spiked with the metabolite of interest [81,83,87,88,90,96–100]. The level of the component of interest is therefore suggested to be higher compared to case examples. Ifa et al. [83] described the detection of drugs and explosives in fingermarks, followed by the development of the fingermark ridge pattern using DESI-MS, as depicted in Fig. 4. Kaplan-Sandquist et al. [98] investigated the presence of drug ions in fingermarks residues using MALDI-time-of-flight (TOF)-MS. Tablets of different types of drugs were placed between two fingers to mimic the process of drug ingestion. Also broken tablets were handled by the volunteers before placing their fingermarks. The target drug ions could not be detected in contaminated fingermarks. Probably, the coating of the tablets limits the transfer of detectable amounts of the drugs to the fingermark residue. Also, the amount of drugs present in the fingermarks exposed to broken tablets did not result in a positive signal, whereas direct handling of the analyte powders resulted in a good detection of the drug in the fingermark residue [98]. In conclusion, the detection of drugs and their metabolites in fingermarks has been extensively investigated, presenting a large variety of different techniques available to detect these components in fingermarks. If fingermarks have been developed prior to chemical analysis, techniques such as SALDI-MS, SIMS, SERS and immunolabelling are available. An advantage of Raman spectroscopy (RS) is that after development and lifting it is still possible to spectroscopically detect chemical components in fingermark residues. However, the choice of lifting tape is important, as some lifting tapes have a high background fluorescence that may interfere with the Raman spectrum [96,101]. It must be noted that the fingermarks used in these studies were fresh fingermarks and that in most cases the best suitable surface and/or conditions were chosen suitable for the used technique. To make one of these techniques directly applicable to the forensic field, a large validation study is required. Based on the requirements of the forensic investigator the best suitable method can be selected. 3.7. Other foreign materials: explosive- and gunshot residues The presence of components related to explosive- and gunshot residues in fingermarks can provide important additional information on handlings of the donor. The relation between criminal activity and identity is helpful information in the forensic investigation. In a controlled firing experiment, fingermarks of donors were investigated on the presence of compounds related to gunshot residue (GSR). An increased level of nitrite and nitrate was found in these fingermarks, however the typical GSR species cyanate was only found to be mildly increased in fingermarks from donors who fired guns compared to control

7

fingermarks. Nitrite and nitrate may also originated from an increased sweat excretion during the activity of firing and should therefore be interpreted with special care [102]. Detection of components specific for GSRs is difficult, since none of the elements present in GSR are unique to the handling of firearms only [103]. Additionally, GSR is not that persistent on hands. In an extensive review by Saverio and Margot [104] they described that GSR particles larger than 10 μm disappeared within two hours after firing. Smaller particles of less than 3 μm could be observed after 2 h or more. Washing with water and soap removed the GSR completely from the hands. Techniques that may provide information on the presence of GSR and explosives are DART-MS, highperformance liquid chromatography (HPLC), near infrared spectroscopy (NIR) in combination with HSI, SALDI-TOF-MS, C+ 60 secondary ion mass spectrometry (SIMS) and different types of spectroscopy [79,81, 89,98,105–110]. The main drawback of these studies is that the fingermarks used were spiked with the component of interest, which can contain a higher amount of the component and can thus deviate from real case examples. Fingermarks contaminated with explosive residues, which can be associated with a planned bombing or actual bombing, are crucial in the forensic investigation [13]. The detection of explosive residues in fingermarks, such as 2,4,6-trinitrotoluene (TNT), RDX, pentaerythritol nitrate (PETN), chlorate and nitrate is possible and can be achieved using different techniques [13]. Explosive particles of around 20 μm can be visualized and analysed using ATR-FTIR, Mou et al. [111] was able to distinguish natural fingermarks from fingermarks contaminated with particles of explosive material. Visual examination of the fingermarks with the microscope did not result in discrimination, because the particles normally present in fingermarks have the same morphology as the particles present in explosives. Although after detection the particles are compressed and larger than before the measurement, ATR-FTIR affects the fingermark minimally, since the ridge pattern is not disturbed as the diameter of the ATR crystal is narrower than the ridges [111]. RS can be used to detect tiny explosive particles in fingermarks residues, such as RDX, HMX, PETN and ammoniumnitrate [112]. Fig. 5 shows the detection of potassium nitrate, urea nitrate and dinitrotoluene in fingermarks using Raman spectroscopy. In conclusion, foreign substances can include a whole set of different compounds originating from various sources, including gunshot residues and explosive particles and can be detected and analysed using a wide range of techniques. However, more research should be performed on which compounds have enough discriminative power to use for donor profiling and which compounds provide interesting knowledge on the handlings of the donor.

Fig. 4. A: DESI image of distribution of cocaine on a spiked latent fingermark deposited on glass. B: Computer generated fingermark obtained from the DESI image A. Important fingermark details are indicated in B with red circles. This figure has been adapted and reprinted from Ifa DR, Manicke NE, Dill, AL, Cooks, RG: Latent Fingerprint Chemical Imaging by Mass spectrometry. Science 2008, 321 (5890): 805 [83]. Copyright 2008, with permission from Science.

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Fig. 5. Response corrected Raman spectra of (a) potassium nitrate, (b) urea nitrate and (c) 2,4 dinitrotoluene and in the right side, images of the individual particles, obtained from the fingermark residues of the different compounds, which were used for measuring the corresponding spectra with the compact Raman spectrometer in 10 s. Applied Physics B, Lasers and Optics 2013: 511–518 [109]. Copyright 2012, with permission from Springer.

3.8. Health information The chemical composition of a fingermark will not only be interesting for the forensic field, but can also be of special interest for the medical field. Since fingermarks are mainly composed of sweat originating from the body, including a large variation of different lipids, proteins, peptides and metabolites, information on the health status of individuals is assumed to be reflected in the chemical properties of the fingermark. The use of fingermarks as diagnostic tool can be extremely valuable, since the sample material is obtained non-invasively. However, up to now, information on the health status of an individual based on fingermarks residue is not possible. The use of sweat is currently investigated by different research groups for non-invasive diagnostics [113–115]. A diagnostic sweat test for new-borns is available that is able to diagnose for cystic fibrosis based on the osmolarity and sodium levels [114]. Different possible biomarkers that are related to diseases can be found in sweat, however the diagnostic value has not yet been investigated [114]. Only when it becomes clear what the biomarkers tell us can we try to detect them in fingermarks and prove their value as diagnostic tools. As described in the previous sections, various components, originating from endogenous and exogenous sources, can be detected in fingermarks, that may provide additional information about the donor of the fingermark. Now that the techniques available to extract donor profiling information from fingermarks have been brought to the attention of the forensic field, research can focus on readying those techniques that have the greatest potential to impact forensic casework. In Table 1 a schematic overview of the techniques that can be used to obtain specific types of donor information is given. We expect, however, that much more information is present in fingermarks than has been presented.

4. Discussion Although a number of techniques exist that allow for the detection of specific components in fingermarks, none of these techniques are currently used during crime scene investigations. As described in this review, additional interesting information of the donor can be obtained from fingermarks besides the ridge pattern, such as gender, and whether they came in contact with drugs or explosives. Many analytical techniques are available, each with its own advantages and disadvantages hampering selecting one of these as ultimate technique for donor profiling of fingermarks. Furthermore, the quality of visualization of the ridge pattern also depends on the selected technique, adding an additional complication in the decision making process. In Table 1, a schematic

overview is presented, discussing the techniques including their advantages and disadvantages. Besides the ridge pattern, the DNA present in fingermark might also reveal the identity of the donor. However, to allow DNA analysis, the fingermark needs to be dissolved, which is destructive to the ridge pattern and prevents further chemical analysis on these samples. As shown in Table 1, most of the presented techniques that acquire donor profiling information from fingermarks are non-destructive or minimally destructive. Hence, after analysing the chemical profile of the fingermarks to extract important intelligence information, such as gender, age or blood type, the genetic material present in the fingermark is still available for DNA analysis. There are several reasons why none of these techniques are implemented in the forensic field. One of these reasons is that there is still a gap between forensic experimental research and forensic practice. Another reason is that none of the techniques has been properly validated for fingermark applications. Only a limited amount of fingermarks were investigated to prove the concept of the technique and to demonstrate that specific components can be identified using the described techniques. Besides the sample size, other variations can affect the initial composition of the fingermark as well as the ageing of fingermarks and therefore the detection of certain components. A good overview of the different factors that influence the ageing of fingermarks has been recently described in a review by Cadd et al. [126]. Donor characteristics, deposition conditions, substrate nature, environmental conditions and contaminations can all influence the chemical composition of the fingermarks and could influence the ageing and degradation process of the chemical components [18,40,126–128]. Temperature, humidity precipitation and light exposure will all affect the composition of the fingermarks, therefore it is important to investigate the effect of environmental conditions on fingermark residues and whether or under what conditions fingermarks can still be used for donor profiling. Upon ageing of the fingermark chemical components present in the fingermark are exposed to degradation, which might hamper the analysis of specific components in the fingermarks. Future studies should not only focus on fresh fingermarks, but should also include fingermarks that have been aged under different conditions. Additionally, the surface on which fingermarks are left can be a limiting factor. In most studies the most appropriate surface has been studied, whereas in casework all conceivable surfaces are included, including non-porous, porous, coloured and structured surfaces. The different techniques and methods that can be used to retrieve donor profiling from fingermarks have only been tested in population studies with fingermarks from volunteers that were instructed on ‘how’ to place their fingermark and therefore at this point, it is not known how these techniques will act in real case samples. Creating a donor profile will not only give information about the components from intrinsic origin, but also involves contamination from external sources. Contamination originating from for example handshaking might affect the chemical composition of the fingermark and can therefore result in false positive results. Therefore, interpretation of the data should be done with great care. On the other hand, contamination of the fingermark originating from external sources provides also information on the actions of the donor of the trace and can be used to assess testimonies. Very recently it has been shown by Sundar and Rowell [100] that drug components present in the fingermark might be transferred to other fingermarks during dusting of the marks with powder. The hairs of the brush are likely to transfer components from one fingermark to the other. Although the fingermarks in this study were spiked with a relative high concentration of drugs (10 μl of 10 mg/ml, 1 mg/ml and 100 ng/ml), crosscontamination is a serious problem that might affect the initial fingermark [100]. Crime scenes are now only searched for useful fingermarks for the identification process. These fingermarks need to be of good enough quality, concerning the ridge pattern, to use for the crime scene

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Technique

Non-destructive Sample Detection Imaging Surface Compatibility fingermark developers pre-paration limit capability applicability

MALDI-MSI [8,63,80,97,116]

Minimally

Yes

fg

Yes

Non-porous

No, only with special powder that can be used for visualization and as a matrix

SALDI-MS [80,105,118]

Minimally

Yes

ng

Yes

Porous Non-porous

Powder dusting

Limitations - Vacuum conditions - Poor reproducibility - For each application most effective matrix needs to be investigated, time-consuming - Matrix can produce high chemical background interference - Surface influences analysis

Advantages

Donor information

- Can be applied on lifted fingermarks - Portable MS systems are available

-

- Can be applied on lifted fingermarks

-

DART-MS [53,80,119–121]

DESI-MS [83,121–123]

Minimally

No

No

No

ng

ng

No

Yes

Porous Non-porous

Non-porous

ND*

ND

-

- Difficulty with background interference - Detection of explosives not possible in lifted fingermarks - Limited effective mass range (b1 kDa) - High effective mass range

-

SIMS [106,123]

No

No

fg

Yes

Non-porous

Powder dusting

- High vacuum conditions

-

GC–MS [35,40,116]

No

Yes

ng

No

None

ND

- Limited to volatile substances

-

LC-MS [35,40,116]

No

Yes

ng

No

None

ND

- Less sensitive than GC-MS

-

IR [111,125]

Yes

No

ng

Yes

Non-porous

ND

- Difficulty with impure samples. - Not yet possible to analyse whole fingermark at once.

- Can be applied after lifting the fingermarks - Portable

-

RS [84,101]

Yes

No

pg

Yes

Porous Non-porous

Powder dusting

- Not yet possible to analyse whole fingermark at once

- Can be applied on lifted fingermarks - Portable

-

Yes

No



Yes

Porous Non-porous

ND

- Background interference

- Portable

-

Immunolabelling [54,59,64,69,70,75–77]

Yes

Yes

pg

Yes

Porous Non-porous

Powder dusting Ninhydrin Indanedione- Zinc Physical developer Cyanoacrylate

- Background staining - Not applicable at crime scene

- Can be applied on lifted fingermarks

-

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HSI [78,79]

Diet [52] Gender [8] Personal hygiene[63] Lifestyle [14,80,82,90–93,97,98] Foreign substances[98,117] Lifestyle [14,80,82,88,95,100] Foreign substance [105] Diet [53] Lifestyle [53,80,88,121,122] Foreign substances [53,105] Diet [121,122] Lifestyle [83,121,122,124] Foreign substances[122] Personal hygiene [106] Foreign substances [106] Age[33,34] Gender [12,32,34–36] Personal hygiene [12,62] Lifestyle[122] Foreign substances [122] Diet [11] Lifestyle [11,86,94,122] Age [10,44,45,47] Diet[14] Personal hygiene [61] Lifestyle [81] Foreign substances [110,111,117,125] Diet [101] Lifestyle [74,84,87,89,96,101] Foreign substances [84,89,109,112,117] Age [44] Diet[78] Lifestyle [78] Foreign substances [79,81] Blood group typing [59] Lifestyle [60,66–70,74,75,77]

A. van Dam et al. / Science and Justice xxx (2015) xxx–xxx

Please cite this article as: A. van Dam, et al., Techniques that acquire donor profiling information from fingermarks — A review, Sci. Justice (2015), http://dx.doi.org/10.1016/j.scijus.2015.12.002

Table 1 Schematic overview of the different techniques with their advantages and disadvantages. *ND=not determined

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investigation. The amount of traces included in the crime scene investigation is therefore limited. Donor profiling can be applied to each type of fingermark trace, including smudged fingermarks, distorted fingermarks and partial fingermarks. Of course, the techniques are not limited to fingermarks, also other traces, including minimal contact traces, smears, blood, sperm and saliva can be analysed with the described techniques. The choice of technique depends on the type of information requested by the investigator. When no prior knowledge is available, the most interesting information will be the gender and age of the donor. If knowledge about the age of the donor is questioned, GC-MS and FTIR seem to be promising techniques since age estimation of the donor is not investigated with the other described techniques. Given that GCMS is a destructive technique and will destroy the fingermark for further analysis, FTIR will probably be preferred over GC-MS. Gender determination is currently possible only with MALDI-MS. A few possible gender biomarkers are described in literature, including DCD-1L, SSL29 and LEK-45, other techniques, such as immunolabelling, can probably also be used for gender determination [8]. Additionally, a combination of techniques and/or methods can be used to obtain more reliable results. Combining the chemical information with the ridge pattern will not only increase the amount of information obtained from the fingermark, but may also lead to other insights since the components in the fingermark combined with the location of the fingermark may provide intelligence about, for instance, the order of events. When the donor of the fingermark is known, information such as drug usage or handling of certain items, including explosives, might be interesting, which may help in the verification or falsification of testimonies. Almost all described techniques can be used to detect drugs and drug metabolites in fingermarks. The choice of technique will depend on many factors such as the costs, time-effectiveness, destructiveness, sensitivity, specificity and reliability of the method. The circumstances in which the trace has been found, including the surface and the visibility of the trace, will also affect the choice of technique, since not all techniques can be used at the crime scene or directly on fingermarks that have been developed with a fingermark developer. Portable systems are preferred over laboratory work, since fingermarks that are directly analysed at the crime scene are less likely to be exposed to contamination, get lost or get damaged during transport. However, moving the described techniques from the laboratory to the crime scene will be challenging. Portable systems are available, for instance portable HIS, MS and portable RS. In many studies spiked fingermarks were used, but the amount of compounds of interest is likely to be higher in the spiked fingermarks than in real case examples. Additionally, in crime scene investigations fingermarks can be composed of a large variety of components, including components that might hamper the chemical analysis. Further research should focus on fingermarks that contain naturally excreted components. Logically, implementation of the techniques will not only provide additional information from traces, but will also mean additional work. Not only more manpower is necessary, but also implementation of the techniques will involve considerable costs depending on the chosen technique. Although the costs of new equipment may be high, collaboration with academic institutes that already have these instruments might offer profitable solutions as well as insights into the development of instruments and techniques. Depending on the type of crime, the impact, volume and profile of the crime, one can decide to perform additional analysis on the fingermarks. At this moment, a few steps need to be taken, before donor profiling of fingermarks can be implemented in the forensic field. For instance, the effect of environmental conditions, ageing and the effect of contamination from external sources/humans are important factors that might influence the interpretation of the data. Therefore, it is important that future studies focus on these important aspects to bridge the gap between research and the forensic field and to reach

the final goal, implementation of these techniques and applications in the forensic field. 5. Conclusion Fingermarks contain a lot of information that can provide additional intelligence on the donor of the mark, which can be especially helpful when the identity of the donor is unknown and/or when fingermarks are smudged, distorted and not useful for identification. Donor profiling information from fingermarks includes information on gender, age of the donor, contact with foreign substances and lifestyle. Different techniques are available that can be used to obtain these types of information. Depending on what kind of information is questioned and in which specific circumstances the fingermark is found, the best suitable technique can be chosen. 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