Evaluation of extraction techniques for the forensic analysis of human scalp hair using gas chromatography/mass spectrometry (GC/MS)

Preliminary research using on-line supercritical fluid extraction/gas chromatography-mass spectrometry (SFE/GC-MS) has shown that the natural and artificial surface components of human scalp hair are reproducible and differentiable. Therefore, these components may be useful for individualization or determining demographic characteristics or both. However, it is not known how the efficiency and selectivity of on-line SFE/GC-MS compares to other extraction methods. In this study, ultrasound, Soxhlet, and pressurized-fluid extraction were used to extract 1 mg to 1.3 g portions of a composite hair sample taken from an Asian male between the ages of 10 and 18. Percent extractables ranged from 0.9% to 5.6%, depending on the solvent used, and tended to increase with solvent polarity. Chemical analysis using GC/MS showed that the extracts contained large proportions of free fatty acids, squalene, cholesterol, and various wax esters. Finally, comparisons to SFE/GC-MS showed that this method possesses adequate efficiency, no observable differences in selectivity, and greater potential for miniaturization.     

Laser desorption/ionization time-of-flight mass spectrometry of triacylglycerols and other components in fingermark samples

The chemical composition of fingermarks could potentially be important for determining investigative leads, placing individuals at the time of a crime, and has applications as biomarkers of disease. Fingermark samples containing triacylglycerols (TAGs) and other components were analyzed using laser desorption/ionization (LDI) time-of-flight mass spectrometry (TOF MS). Only LDI appeared to be useful for this application while conventional matrix-assisted LDI-TOF MS was not. Tandem MS was used to identify/confirm selected TAGs. A limited gender comparison, based on a simple t-distribution and peaks intensities, indicated that two TAGs showed gender specificity at the 95% confidence level and two others at 97.5% confidence. Because gender-related TAGs differences were most often close to the standard deviation of the measurements, the majority of the TAGs showed no gender specificity. Thus, LDI-TOF MS is not a reliable indicator of gender based on fingermark analysis. Cosmetic ingredients present in some samples were identified.     

Changes in the lipid composition of latent fingerprint residue with time after deposition on a surface

The work described in this report was focused on generating increased knowledge of fingerprint chemistry, particularly the composition of a latent fingerprint at the time it is deposited, and the chemical changes in lipid components that occur over time. Fingerprints from five male donors (aged 25-34 years) were collected and aged under controlled conditions. The prints were then sampled at set intervals, solvent extracted with dichloromethane, co-derivatized with MSTFA and analyzed by gas chromatography-mass spectrometry (GC-MS). It was shown that there was loss of squalene from prints stored in the light or in the dark. Loss was more rapid in the light, with squalene in prints from some donors not detected after 9 days storage. For these same donors, squalene was still detected after 33 days storage in the dark, but at much lower levels than in fresh prints. For saturated fatty acids (tetradecanoic, palmitic and stearic acid) there was a trend towards an increase in levels of these substances during storage (up to about 20 days) followed by a decrease back to original levels or below. This was the case for samples stored in the light or in the dark. For palmitoleic acid, a similar trend was seen. For oleic acid, this trend was seen for samples stored in the dark. For samples stored in the light the general trend was a decrease in level over the storage period (up to 33 days).     

指纹乳突纹线密度的性别差异初探

目的针对指纹纹线密度在不同性别间是否存在差异进行了研究和探索。方法实验对80个（其中男女各40人）中国汉族人十指纹线密度进行计数,将所得数据通过SPSS软件进行统计分析。结果指纹纹线密度均值男性少于14.5线/25mm2,女性大于等于15线/25mm2。结论男女之间指纹密度存在差异;女性指纹纹线密度较男性高。     

The impact of lipid degradation on fingerprint quality on fired firearm cartridges

The recovery of identifiable fingerprints from fired cartridge cases is challenging. Therefore, the characterization of chemical modifications and their effects on fingerprint integrity post-firing is essential. In this study, the primary fingerprint lipids, including myristic acid, pentadecanoic acid, palmitoleic acid, palmitic acid, stearic acid, squalene, and cholesterol in fired and unfired cartridges, were extracted with acetonitrile, followed by derivatization using N,O-Bis(trimethylsilyl)trifluoroacetamide with 1% trimethylchlorosilane (BSTFA/1%TMCS). Squalane was used as the internal standard, and all quantifications were performed using gas chromatography coupled with mass spectrometry using a triple-quadrupole mass filter. All lipids identified in the unfired cartridges were also detected in the fired cartridges, and statistical analysis using Student's t-test and F tests was performed with a 95% confidence level. The concentration of lipids in the unfired cartridges was found to be similar to that detected in the fired cartridges, except for squalene, the recovery of which was 28% lower in the fired cartridges.     

Toward multidimensional information: A derivatization-free UHPLC-QqQ MS/MS method for amino acid components of fingerprint

The analysis of fingerprint chemical composition is a meaningful way to excavate the multidimensional information of fingerprint, including the donor profiling information and the age of a fingerprint, which broadens the evidential values of fingerprint, especially for the partial and distorted fingerprint. But the research remains still in the pilot phases or is ongoing. Amino acids are the dominant organic substances in latent sweat fingerprint and influenced by many donor factors. Hence, their content reflects personal information of donors. Forensic science will be revolutionized if suspects can be individualized by their amino acid content. The diverse nature, distinct physicochemical properties, and ultra-micro levels of amino acids present in fingerprints make it hard to detect. A high sensitivity method for detecting and quantifying multiple amino acid components is required. UHPLC-QqQ MS/MS offers high sensitivity, high separation, simultaneous multicomponents detection, and no derivatization, making it an ideal method for detecting and analyzing amino acids in fingerprints. Therefore, in this study, we propose and validate an efficient UHPLC-QqQ MS/MS method for the extraction and analysis of 13 amino acids from fingerprint. We compared the results of amino acids of 10 different substrates and found that the inherent amino acids in most porous substrates would have been extracted along with the fingerprint amino acids, making them unsuitable for quantitative amino acid analysis. Instead, plastic sheets are ideal substrates for laboratory studies. Then, extensive experiments were conducted among 30 donors for multidimensional information analysis. The type of samples analyzed were eccrine-rich fingerprints. A Binary Logistic Regression (BLR) model was developed, and the female and male donors were successfully differentiated by amino acids in fingerprints. Two other mathematical models were also developed to verify the accuracy, and all three different mathematical models were able to identify donors of different genders with over 90% accuracy. This demonstrates that amino acids have the potential to provide more information for donors as metabolic markers. In the future, we will conduct a series of experiments to analyze more multidimensional information for individual identification by amino acid content in the fingerprint.     

Chemical Differences Are Observed in Children’s Versus Adults’ Latent Fingerprints as a Function of Time*

Abstract: The identification of aged latent fingerprints is often difficult, especially for those of children. To understand this phenomenon, the chemical composition of children’s versus adults’ latent fingerprints was examined over time using Fourier transform infrared microscopy. Hierarchical cluster analysis revealed that children’s and adults’ prints were distinguishable for up to 4 weeks after deposition, based on differences in sebum composition. Specifically, adults had a higher lipid content than children, but both decreased over time, attributable to the volatility of free fatty acids. The aliphatic CH₃, aliphatic CH₂, and carbonyl ester compositions changed differently in adults versus children over time, consistent with higher cholesterol and cholesteryl esters in children’s prints and wax esters and glycerides in adults’ prints. Thus, fingerprint composition changes with time differently in children versus adults, making it a sensitive metric to estimate the age of an individual, especially when the age of the print is known.     

A further study to investigate the detection and enhancement of latent fingerprints using visible absorption and luminescence chemical imaging

This study investigated the application of chemical imaging to the detection of latent fingerprints using the Condor macroscopic chemical imaging system (ChemImage Corp., Pittsburgh, USA). Methods were developed and optimised for the visualisation of untreated latent fingerprints and fingerprints processed with DFO, ninhydrin, cyanoacrylate, and cyanoacrylate plus rhodamine 6G stain. The results obtained with chemical imaging were compared to the detection achieved using conventional imaging techniques. The Condor significantly improved the detection of many prints, especially those that might be considered poor quality or borderline prints. Prints on newspaper treated with ninhydrin and DFO, and prints on white and yellow paper treated with ninhydrin, benefited the most from chemical imaging detection. In many cases, fingerprints undetectable using conventional imaging techniques could be visualised with chemical imaging. Ridge detail from untreated prints on yellow paper was also detected using the Condor. When prints of high quality were examined, both detection techniques produced quality results. The results of this project demonstrate that chemical imaging offers advantages over conventional visualisation techniques when examining latent fingerprints, especially those that would be considered difficult, such as weak prints or prints on surfaces that produce highly luminescent backgrounds. Standard testing procedures for the detection and enhancement of fingerprints by chemical imaging are presented and discussed.     

Determining the order of deposition of natural latent fingerprints and laser printed ink using chemical mapping with secondary ion mass spectrometry

Time-of-flight secondary ion mass spectrometry (ToF-SIMS) chemical mapping was used to investigate the order of deposition of natural latent fingerprints and laser printed ink on paper. This feasibility study shows that sodium, potassium and C₃H₅ positive ions were particularly abundant endogenous components of the natural fingerprints and also present in the paper examined, but were mostly absent in the laser printed ink. Mapping of these ions enables the observation of friction ridges from latent prints on the ink surface, only when a fingerprint was deposited above the layer of ink. As a demonstration of proof of concept, blind testing of 21 samples from three donors resulted in a 100% success rate. The sensitivity of this technique was investigated within this trial through the examination of up to fifth depletion fingerprints and ageing of up to 28 days. Migration of fingerprint and paper components to the ink surface, although observed with increased ageing time, was not found to compromise determination of the deposition sequence.     

Fingerprint enhancement revisited and the effects of blood enhancement chemicals on subsequent profiler Plus fluorescent short tandem repeat DNA analysis of fresh and aged bloody fingerprints

This study was aimed at determining the effect of seven blood enhancement reagents on the subsequent Profiler Plus fluorescent STR DNA analysis of fresh or aged bloody fingerprints deposited on various porous and nonporous surfaces. Amido Black, Crowle's Double Stain. 1,8-diazafluoren-9-one (DFO), Hungarian Red, leucomalachite green, luminol and ninhydrin were tested on linoleum, glass, metal, wood (pine, painted white), clothing (85% polyester/15% cotton, 65% polyester/35% cotton, and blue denim) and paper (Scott 2-ply and Xerox-grade). Preliminary experiments were designed to determine the optimal blood dilutions to use to ensure a DNA typing result following chemical enhancement. A 1:200 blood dilution deposited on linoleum and enhanced with Crowle's Double Stain generated enough DNA for one to two rounds of Profiler Plus PCR amplification. A comparative study of the DNA yields before and after treatment indicated that the quantity of DNA recovered from bloody fingerprints following enhancement was reduced by a factor of 2 to 12. Such a reduction in the DNA yields could potentially compromise DNA typing analysis in the case of small stains. The blood enhancement chemicals selected were also evaluated for their capability to reveal bloodmarks on the various porous and nonporous surfaces chosen in this study. Luminol. Amido Black and Crowle's Double Stain showed the highest sensitivity of all seven chemicals tested and revealed highly diluted (1:200) bloody fingerprints. Both luminol and Amido Black produced excellent results on both porous and nonporous surfaces, but Crowle's Double Stain failed to produce any results on porous substrates. Hungarian Red, DFO, leucomalachite green and ninhydrin showed lower sensitivities. Enhancement of bloodmarks using any of the chemicals selected, and short-term exposure to these same chemicals (i.e., less than 54 days), had no adverse effects on the PCR amplification of the nine STR systems surveyed (D3S 1358, HumvWA, HumFGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820) or of the gender determination marker Amelogenin. The intensity of the fluorescent signals was very similar and the allele size measurements remained constant and identical to those of untreated bloody fingerprints. No additional background fluorescence was noted. Continuous exposure (for 54 days) to two of the seven enhancement chemicals selected (i.e., Crowle's Double Stain and Hungarian Red) slightly reduced the amplification efficiency of the longer STR loci in profiles of fresh and 7 to 14-day-old bloodprints. This suggests that long-term exposure to these chemicals possibly affects the integrity of the DNA molecules. This study indicates that significant evidence can be obtained from fresh or aged bloody fingerprints applied to a variety of absorbent and nonabsorbent surfaces which are exposed to different enhancement chemicals for short or long periods of time. It also reaffirms that PCR STR DNA typing procedures are robust and provide excellent results when used in concert with fluorescence-based detection assays after fingerprint identification has taken place.     

Threat mail and forensic science: DNA profiling from items of evidence after treatment with DFO

Two cases of threatening letters with their accompanying envelopes were received to the Division of Forensic Identification unit of the Israel Police. The envelopes, including the stamps, and the letters were initially examined for latent fingerprints by the DFO reagent, known to cause degradation of DNA. Although no latent fingerprints could be visualized on any of the items, the biology laboratory using organic DNA extraction, was successful in defining genetic profiles from all the items employing six STR loci, even after treatment with DFO. In a controlled experiment, a known donor attached a stamp, by licking, to an envelope. This item was treated with DFO and then profiled using STR loci. The results showed that previous DFO treatment on the control stamp before DNA analysis had no negative effects on obtaining the DNA profile of the known donor using STR loci.     

DNA recovery after sequential processing of latent fingerprints on copy paper

Forensic examiners must determine whether both latent fingerprint development and DNA profiling can be performed on the same area of an evidence item and, if only one is possible, which examination offers the best chance for identification. Latent fingerprints can be enhanced by targeting different components of fingerprint residues with sequential chemical treatments. This study investigated the effects of single-reagent and sequential latent fingerprint development processes on downstream DNA analysis to determine the point at which latent fingerprint development should be stopped to allow for DNA recovery. Latent fingerprints deposited on copy paper by one donor were developed using three sequential processes: 1,8-diazafluoren-9-one (DFO) → ninhydrin → physical developer (PD); 1,2-indanedione-zinc (IND-Zn) → ninhydrin → PD; and IND-Zn → ninhydrin → Oil Red O (ORO) → PD. Samples were examined after the addition of each chemical treatment. DNA was collected with cotton swabs, extracted, quantified, and amplified. DNA yields, peak heights, number of alleles obtained, and percentage of DNA profiles eligible for CODIS upload were examined. DNA profiles were obtained with varying degrees of success, depending on the number and type of treatments used for latent fingerprint development. The treatments that were found to be the least harmful to downstream DNA analysis were IND-Zn and IND-Zn/laser, and the most detrimental treatments were DFO, DFO/laser, and PD. In general, as the number of treatments increase, the opportunities for DNA loss or damage also increase, and it is preferable to use fewer treatments when developing latent fingerprints prior to downstream DNA processing.     

A novel chitosan/tripolyphosphate/L‐lysine conjugates for latent fingerprints detection and enhancement

Most chemical and physical methods employed in visualizing latent marks have shown detrimental effects on human health and, therefore, some research groups have directed their attention to the utilization of various (bio)polymers with the aforementioned purpose. Although chitosan is widely used in medicine, pharmacy, food, and drug delivery systems, there are very few studies that address this biopolymer utilization in forensic applications, such as the detection of latent fingerprints. We used chitosan crosslinked with sodium tripolyphosphate and conjugated with L‐lysine to enhance the visualization of latent fingerprints, due to its ability to interact with fingerprint sweat residues. These conjugates were prepared using different (w/w) amounts of chitosan and tripolyphosphate (6/1; 4/1; 1/1; 1/4; and 1/6), and those with the most promising results (i.e., 6/1 formulation) were investigated in detail. Fourier transform infrared (FT‐IR) spectroscopy confirmed interactions between components of the systems. Optical microscopy and scanning electron microscope (SEM) analysis showed that prepared powder formulations were uniform in size and confirmed that chitosan/tripolyphosphate/lysine conjugates bind easily to the sweat and lipid residues present in the latent fingerprints. The testing of prepared conjugates demonstrated the potential of these systems as bio‐based powder substitution for commercially available powders.     

Carbon‐Based Fingerprint Powder as a One‐Step Development and Matrix Application for High‐Resolution Mass Spectrometry Imaging of Latent Fingerprints

Carbon‐based materials are often used as matrices for matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐MS) and its imaging (MALDI‐MSI). However, researchers have refrained from using carbon‐based fingerprint powder (CFP) as a matrix due to high background and contamination. In this work, the compatibility of CFP is reevaluated with MALDI‐MSI using a high‐resolution mass spectrometer (HRMS) and compared to traditional organic matrices. Relevant fingerprint compounds were easily distinguished from carbon cluster peaks when using HRMS. For fair comparison, half of a fingerprint was dusted with CFP while the other half was dusted with traditional organic matrices. All compounds studied had comparable, or higher, signal‐to‐noise (S/N) ratios when CFP was used as the matrix. Additionally, chemical image qualities closely followed the trend of S/N ratios. CFP proved to be an effective one‐step development and matrix application technique for MALDI‐MSI of latent fingerprints, when carbon cluster peaks are well separated by a HRMS.     

Initial Results on the Composition of Fingerprints and its Evolution as a Functionof Time by GC/MS Analysis

Abstract: Determining the time since deposition of fingermarks may prove necessary to assess their relevance to criminal investigations. The crucial factor is the initial composition of fingermarks, because it represents the starting point of any aging model. This study mainly aimed to characterize the initial composition of fingerprints, which show a high variability between donors (inter‐variability), but also to investigate the variations among fingerprints from the same donor (intra‐variability). Solutions to reduce this initial variability using squalene and cholesterol as target compounds are proposed and should be further investigated. The influence of substrates was also evaluated, and the initial composition was observed to be larger on porous surface than nonporous surfaces. Preliminary aging of fingerprints over 30 days was finally studied on a porous and a nonporous substrate to evaluate the potential for dating of fingermarks. Squalene was observed to decrease in a faster rate on a nonporous substrate.     

间接免疫酶组化法检测人指纹ABO血型

建立一套测定人指纹 ABO血型的方法体系。运用间接免疫酶技术和免疫印迹技术 ,对指纹胶纸提取的指纹、银粉显现提取的指纹、直接转移至硝酸纤维素膜 (NC膜 )上的指纹、银粉显现转移至 NC膜上的指纹、“5 0 2”胶熏显转移至 NC膜上的指纹等进行 ABO血型检测。上述 5种方法采集的 2 12枚指纹 ,绝大部分能正确地检出 ABO血型 ,检出率为 90 %～ 93.8% ;未能检出的样本为非分泌型人指纹。应用间接免疫酶技术及免疫印迹技术测定人指纹的 ABO血型的方法具有特异性强、准确可靠、检测方便等优点 ,在法庭科学中易推广应用 ,并能发挥重要的作用     

Effect of Aging and Surface Interactions on the Diffusion of Endogenous Compounds in Latent Fingerprints Studied by Mass Spectrometry Imaging,

The ability to determine the age of fingerprints would be immeasurably beneficial in criminal investigations. We explore the possibility of determining the age of fingerprints by analyzing various compounds as they diffuse from the ridges to the valleys of fingerprints using matrix‐assisted laser desorption/ionization mass spectrometry imaging. The diffusion of two classes of endogenous fingerprint compounds, fatty acids and triacylglycerols (TGs), was studied in fresh and aged fingerprints on four surfaces. We expected higher molecular weight TGs would diffuse slower than fatty acids and allow us to determine the age of older fingerprints. However, we found interactions between endogenous compounds and the surface have a much stronger impact on diffusion than molecular weight. For example, diffusion of TGs is faster on hydrophilic plain glass or partially hydrophilic stainless steel surfaces, than on a hydrophobic Rain‐x treated surface. This result further complicates utilizing a diffusion model to age fingerprints.     

Thermal degradation analysis of amino acids in fingerprint residue by pyrolysis GC-MS to develop new latent fingerprint developing reagents

Conventional development of latent fingerprints is compromised when the prints are decomposed by extreme temperatures, such as those encountered when a weapon cartridge is fired, an improvised explosive device is detonated, and/or in arson cases. Understanding how these extreme temperatures alter the chemical and physical properties of latent fingerprint residue could aid in the discovery of a reagent that could effectively develop these decomposed fingerprints. To mimic scenarios where fingerprints may be exposed to high heat conditions, standards of the five most abundant amino acids in fingerprint residue as well as extracted fingerprint residue were pyrolized under controlled conditions. Compounds identified as pyrolytic decomposition products were 3,6-dimethylpiperazine-2,5-dione (from alanine), maleimide, and 2,5-furandione (from aspartic acid). The pyrograms and selected ion traces show these products to hold promise as indicators of decomposed fingerprint residues and, therefore, may serve as good candidate substrates for a developing reagent.