On the discrimination between facial creams of different brands using Raman Spectroscopy and partial least squares discriminant analysis for forensic application

Determining the origin of cosmetic traces is an important aspect of forensic investigations, that helps linking a suspect to a crime. Such type of evidence can help further narrow down the undergoing investigations. This paper reports the first use of Raman Spectroscopy (RS) coupled with the exploratory principal component analysis (PCA) and supervised partial least squares-discriminant analysis (PLS-DA) in facial creams. 40 facial cream samples of 8 different brands were studied in this work. Preliminary assessments through visual inspection of their Raman spectra revealed the presence of oxides, titanium dioxide, castor seed oil, and beeswax. Also, the peaks of alkyne groups were indicative of the presence of talc or mica compounds. The exploratory PCA correctly segregated the samples into 8 clusters and the supervised PLS-DA model correctly classified them into 8 classes. Further evaluation of the performance of the trained PLS-DA model resulted in perfect classification shown by the receiver operating characteristic (ROC) curves. The PLS-DA model also resulted in 100% accuracy of correctly assigning the brand on the face wipes on each of the five substrates viz. cotton, dry and wet tissue paper, nylon substrate, and polyester. This validation was done treating these samples as unknowns. The study has a potential for use under actual forensic casework conditions.     

A comprehensive ATR-FTIR spectroscopic analysis for the identification and differentiation of lip balms

Lip balm may be encountered as physical evidence in cases involving sexual assaults, homicides, and kidnappings. Lip balm can be used as corroborative evidence by providing a potential link between the victim, accused, and the crime scene. For lip balms to be used as evidence, it is important to understand the diversity and their aging process under different conditions. Therefore, in this study, ATR-FTIR spectroscopy in conjunction with chemometric tools such as principal component analysis (PCA) and linear discriminant analysis (LDA) has been used for the objective identification and differentiation of 20 brands of lip balms. Moreover, lip balms on different substrates and wearing effects over time were also investigated. The results show that the PCA-LDA training accuracy was 92.5%, whereas the validation accuracy comes out to be 83.33%. A blind study using pristine samples was also performed which resulted in 80% PCA-LDA accuracy. PCA-LDA prediction of samples on various substrates showed a higher chemometric prediction accuracy for nonporous substrates (glass, plastic, and steel), than for porous substrates (cotton cloth, cotton swab stick, dry tissue paper, and white paper) for samples kept in room temperature and under sunlight for 15 days. The substrate study showed that the samples from various substrates could effectively generate respective spectra which can help in brand-level identification even after several days. The present method demonstrates a potential for lip balm samples to be used in forensic casework applications.     

Rapid and non-destructive approach for characterization and differentiation of sealing wax using ATR-FTIR spectroscopy

Sealing wax is used for maintaining the integrity and authenticity of a document or physical evidence. Any tampering with the seal calls into question the overall integrity and authenticity of the tangible evidence or document. In these circumstances, determining the authenticity of the sealing material (physical and chemical) becomes imperative. In this study, ATR-FTIR spectroscopy supported by chemometrics has been used to differentiate sealing wax samples belonging to 12 different brands available across India. All the samples were first melted, cooled, and then analyzed using ATR-FTIR spectroscopy in the mid-infrared region (4000–600 cm⁻¹). The obtained spectra were first examined visually for the presence of different functional groups. Principal component analysis (PCA) and principal component analysis-linear discriminant analysis (PCA-LDA) were employed to analyze the sample clustering patterns and to categorize them into their respective groups, respectively. For classification, a PCA-LDA training model was applied, and it demonstrated 95.83% accuracy. The validation test resulted in an accuracy of 83.33%. PCA-LDA model offered 100% accurate prediction for samples on various substrates, including cloth, cardboard, and paper. A blind study was also performed using five unknown samples, which were accurately classified into their respective groups. PCA-LDA model will be helpful in providing investigative leads by linking a questioned sealing wax sample with its respective group.     

Near-infrared spectroscopy combined with chemometrics to classify cosmetic foundations from a crime scene

Cosmetic smears are a form of trace evidence that can link the crime scene, suspects, and victims. Foundation and lipstick are the most common sources of cosmetics that can easily smear, with most current research focused on the evidential analysis of lipsticks. This research aims to create a database of cosmetic foundations on different materials and to access the robustness of using Near-infrared with chemometrics as a non-destructive technique to identify unknown samples collected from a crime scene. Small amounts of six shades of three brands of foundations were smeared on clothing materials, which were then analysed with a combination of Near-infrared with chemometric analysis. Principle component analysis (PCA) was used to reduce data dimensionality and explore potential patterns in sample separation and Linear Discriminant Analysis (LDA) was utilised to assign unknown samples to one of the established classes. The selected techniques proved to be promising for database construction and as a preliminary method of analysis, with 93% of the spectra being correctly classified. Notably, darker foundation shades were less likely to be correctly classified (90% classified correctly) compared to lighter ones (96.7% classified correctly). This could not be improved with Standard Normal Variate (SNV) data pre-treatment or selecting specific NIR regions. This finding is of particular importance; according to the Crime Survey for England and Wales (year ending March 2020) police recorded sexual offences demonstrated that those in Mixed and Black or Black British ethnic groups were significantly more likely to be a victim of sexual assault compared to White, Asian or Other ethnic groups. It is, therefore, crucial to add a wide range of foundation shades, particularly of darker tones, to the future database.     

Rapid and non-destructive identification of claws using ATR-FTIR spectroscopy–A novel approach in wildlife forensics

Differentiation and identification of Royal Bengal Tiger (Panthera tigris tigris) and Indian Leopard (Panthera pardus fusca) claws is a challenging task in wildlife forensics, due to similarity in their morphology, anatomy and chemical compositions as both the species are closely related to each other genetically. ATR-FTIR spectroscopy, which offers a non-destructive and safe alternative technique to other conventional methods, has been employed in the present work to differentiate claws of Royal Bengal Tiger and Indian Leopard. An attempt has been made to differentiate 31 reference claw samples from 16 different Royal Bengal Tigers, 15 different Indian Leopards, and 10 fake claws using ATR-FTIR spectroscopy supplemented with PCA, PLS-DA, and LDA. PCA could not distinguish the samples of two closely related species among themselves as well as from the fake claws. On the other hand, PLS-DA and LDA models both yielded highly significant classification rate for differentiation among the samples of Royal Bengal Tiger, Indian Leopard, and their fake counterparts. Further, seven blind claw samples that were pretended to be unknown to the analyst of both the species are also examined and identified correctly to their respective groups. The R-Square value obtained for PLS-DA model to differentiate Royal Bengal Tiger, Indian Leopard, and fake claws is 0.99, which is highly significant for predictive accuracy. This study shows that ATR-FTIR spectroscopy with PLS-DA/LDA has a potential to present a rapid, non-destructive, reliable, and eco-friendly approach for the accurate identification and differentiation of Royal Bengal Tiger and Indian Leopard claws.     

The use of Polilight in the detection of seminal fluid, saliva, and bloodstains and comparison with conventional chemical-based screening tests

Biological stains can be difficult to detect at crime scenes or on items recovered from crime scenes. The use of a versatile light source may assist in their detection. The ability of Polilight to locate potential semen, saliva, and blood stains on a range of substrates and at different dilutions was tested. We also tested the use of Polilight in comparison with conventional chemical-based presumptive screening tests such as acid phosphatase (AP), Phadebas, and luminol, often used in casework for detecting potential semen, saliva, and blood stains, respectively. The Polilight was able to locate stains that were not apparent to the naked eye. The color of the material on which a stain is deposited can have an effect on the detectibility of the stain. The Polilight was found to be comparable with the AP and Phadebas tests in terms of its sensitivity. In a comparative study between the AP test and Polilight on 40 casework exhibits, one false-negative result was observed when using the Polilight. On a series of mock casework exhibits it was determined that the Polilight can be used successfully to locate saliva stains for DNA analysis. The sensitivity of luminol for detecting potential bloodstains was greater than that of Polilight; however the Polilight has particular application in instances where a bloodstain may have been concealed with paint. Overall, the Polilight is a relatively safe, simple, noninvasive, and nondestructive technique suitable for use in forensic casework.     

Detection and visualization of human tears using alternate light sources for forensic purposes

The current scientific techniques for locating body fluids focus on quick and effective methodologies for easy and reliable identification. Efficient detection and identification of body fluids play a vital role in establishing the ‘corpus delecti’ of a crime. Non-destructive techniques such as the use of Alternate Light Sources (ALS) have been exploited for crime scene searches over large areas and detection of body fluids such as blood, semen, vaginal secretions, and saliva on a range of substrates. Tears are rarely found but can be considered as potential body fluid evidence due to their unique biochemical and molecular properties. Tears are secreted in response to physical or emotional stimuli. Due to the small volume of secretions, they are often overlooked in the crime scene. Tears may be found on surfaces such as clothing, bedding, tissue, handkerchief, or balaclava. The use of ALS to locate tears on tissue paper and fabric surfaces was tested which were not apparent to the naked eye. Tears stains were successfully detected on surfaces of forensic interest with varying sample ages up to three months with a broad excitation spectrum between 254 nm and 410 nm. Dried stains on tissue paper and fabric substrates were better detected with sharp margins, clear stain pattern visibility, and fluorescence intensity in comparison with moist and fresh stains. Tears stains can hence be detected with the use of ALS and suitable filter combinations under normal conditions and do not require any specific settings to locate them. These findings are suggestive for easy and quick identification of tears on large surfaces and as a presumptive test for forensic casework evidence examination.     

Differentiation of Cosmetic Foundation Creams Using Attenuated Total Reflection Fourier-Transform Infrared Spectroscopy: A Rapid and NonDestructive Approach in Trace Evidence Analysis

Cosmetic foundation creams are encountered as evidentiary material in criminal investigations, particularly in cases related to sexual and physical assault against women. Analyzing foundation cream exhibit is a challenging task as the exhibit is recovered in trace quantity with similar hue. In the present study, ATR FTIR spectroscopy which is a rapid, nondestructive, sensitive, reliable, and safe alternative to other analytical techniques has been used to differentiate 31 cosmetic foundation creams belonging to 23 different brands supported by chemometric methods such as principal component analysis (PCA) and linear discriminant analysis (LDA). The discriminating power of visual analysis is found to be 98.0%, while PCA and LDA further increased the discriminating power to 99.3% and 100%, respectively. The blind test is conducted with three unknown samples (pretended to be unknown to the analyst), which were correctly linked with their respective source. Further, the effect of substrate such as tissue paper (dry and wet) and white cotton cloth during sample analysis are also examined to simulate the actual forensic casework conditions. The stains on substrates could be identified and linked to its parent product as well. The reported method provides significant results for the differentiation of cosmetic foundation creams.     

Simultaneous detection and image capture of biological evidence using a combined 360° camera system with single wavelength laser illumination

Forensic investigators frequently utilise light sources to detect and presumptively identify biological evidence. The instrumentation typically deploys single or multiple wavelength exposures at various intensities, which interact with constituents of biological material, initiating fluorescence or improving contrast between the material and substrate. Documentation using sketches and/or photographic approaches follows detection, which are essential for scene reconstruction. Recent research has demonstrated the simultaneous detection and capture of biological evidence using a 360° camera system combined with an alternate light source exhibiting broad wavelength ranges of light. Single wavelength light sources reportedly offer enhanced sensitivity, due to the increased light intensity and narrower bandwidth of light, although their combined use with a 360° camera system has not yet been explored.Samples of human blood, semen, saliva, and latent fingermarks were deposited on to a variety of substrates. A 360° camera system combined with a laser light source was used to detect and capture the samples. Ten participants were asked to detect the samples on images of the substrates without ground truth knowledge. It was possible to detect and capture biological evidence, although success varied according to substrate colour and light intensity. Advantageously, presumptive screening for biological fluids and the simultaneous location and visualisation of such evidence as part of a 360° panorama of the scene for contextual purposes was permitted. There was no fluorescent response from the fingermarks, although the oblique lighting effects appeared sufficient to aid mark detection in some circumstances. The use of single wavelength illumination clearly facilitates identification of a range of forensically important material. When coupled with a 360-degree camera, this allows for simultaneous identification and recording of such evidence in the context of the whole environment.     

Criminalystic: effectiveness of lysochromes on the developing of invisible lipstick-contaminated lipmarks on human skin. A preliminary study

Latent prints are an important evidence for identification. Nowadays, the technical means, the implementation of image processing techniques and the use of database makes it possible to detect and get information from some prints that seem to be useless at first sight. On the other hand, the possibility of using the print as a DNA source has to be considered, so as to double its identifying value. Human skin is a particularly difficult surface for developing this kind of evidences. Although different methods for locating and developing latent fingerprints on the skin have been already described, it has not been found any method, at the revised bibliography, to obtain and develop invisible lipmarks, that is, lipmarks from protective lipstick, or permanent or long-lasting lipstick. The aim of the work that follows is to determine the effectiveness of several reagents for developing invisible lipmarks on the corpses' skin. Preliminary results show that, under the described experimental conditions, the reagents used, Sudan III, Oil Red O and Sudan Black, are effective for obtaining recent latent lip prints on corpse's skin.     

A Rapid,Confirmatory Test for Body Fluid Identification,

We have developed a technique that allows investigators to confirm the presence of blood, semen, and/or saliva in a crime scene sample. It is a confirmatory test where multiple samples can be processed in less than an hour, and it is potentially portable, permitting samples to be processed at the crime scene. Samples at a scene giving a positive result can be further processed while those failing to do so may be ignored. There is a large and growing backlog of DNA evidence in the USA, slowing down the criminal justice system. This backlog has continued to grow despite an increase in the ability to process evidence faster. This technique uses quantum dot molecular beacons to test for tissue‐specific RNA species, identifying particular body fluids. We have demonstrated the tissue specificity of molecular beacons for blood, semen, and saliva.     

Analysis of the fluorescence of body fluids on different surfaces and times

The use of screening techniques, such as an alternative light source (ALS), is important for finding biological evidence at a crime scene. The objective of this study was to evaluate whether biological fluid (blood, semen, saliva, and urine) deposited on different surfaces changes as a function of the age of the sample. Stains were illuminated with a Megamaxx™ ALS System and photographed with a Canon EOS Utility™ camera. Adobe Photoshop™ was utilized to prepare photographs for analysis, and then ImageJ™ was used to record the brightness values of pixels in the images. Data were submitted to analysis of variance using a generalized linear mixed model with two fixed effects (surface and fluid). Time was treated as a random effect (through repeated measures) with a first-order autoregressive covariance structure. Means of significant effects were compared by the Tukey test. The fluorescence of the analyzed biological material varied depending on the age of the sample. Fluorescence was lower when the samples were moist. Fluorescence remained constant when the sample was dry, up to the maximum period analyzed (60 days), independent of the substrate on which the fluid was deposited, showing the novelty of this study. Therefore, the forensic expert can detect biological fluids at the crime scene using an ALS even several days after a crime has occurred.     

Evaluation of Direct PCR Amplification Using Various Swabs and Washing Reagents

DNA profiles were generated via direct amplification from blood and saliva samples deposited on various types of swab substrates. Each of the six non-FTA substrates used in this research was punched with a Harris 1.2 mm puncher. After 0.1 μL of blood or 0.5 μL saliva, samples were deposited on each of these punches, samples were pretreated with one of four buffers and washing reagents. Amplification was performed using direct and nondirect autosomal and Y-STR kits. Autosomal and Y-STR profiles were successfully generated from most of these substrates when pretreated with buffer or washing reagents. Concordant profiles were obtained within and between the six substrates, the six amplification kits, and all four reagents. The direct amplification of substrates which do not contain lysing agent would be beneficial to the forensic community as the procedure can be used on evidence samples commonly found at crime scenes.     

Visualization of Latent Blood Stains Using Visible Reflectance Hyperspectral Imaging and Chemometrics

The detection of latent traces is an important aspect of crime scene investigation. Blood stains on black backgrounds can be visualized using chemiluminescence, which is invasive and requires a darkened room, or near-infrared photography, for which investigators need to change filters manually to optimize contrast. We demonstrated the performance of visible reflectance hyperspectral imaging (400–720 nm) for this purpose. Several processing methods were evaluated: single wavelength bands, ratio images, principal component analysis (PCA), and “SIMPLe-to-use Interactive Self-modeling Mixture Analysis” (SIMPLISMA). Using these methods, we were able to enhance the contrast between blood stains and 12 different fabrics. On black cotton, blood dilutions were visible with a minimal concentration of 25% of whole blood. The hyperspectral camera system used in this study is portable and wireless, which makes it suitable for crime scene use. The described technique is noncontact and nondestructive, so all traces are preserved for further analysis.     

基于拉曼光谱的血液遗留时间研究与模型预测

目的针对犯罪现场血液遗留时间的判断问题,实验以不同遗留时间的血液为研究对象,建立基于拉曼光谱的血液遗留时间的辨别方法。方法采集遗留时间为0.5h~240h的指尖血液样本,获取其拉曼光谱数据,经校正和平滑处理后,对数据进行归一化。利用RSD值评价光谱的稳定性,选取前10个主成分和6个重要波段分别建立模型,利用预测集测试模型效果。结果相同遗留时间血液的拉曼光谱具有较好的稳定(RSD<0.2),不同遗留时间血液的拉曼光谱在680cm^-1等6个波段有明显的强度变化。前10个主成分建立的PLSR模型,其r²=0.9927。6个重要波段建立的PLSR模型,预测集效果r²=0.8797。结论利用拉曼光谱结合建模分析是一种无损快速的评估血液遗留时间的方法。     

人体生物性物质来源鉴定及法医学应用研究进展

犯罪现场中人体生物性物质来源鉴定在重现犯罪过程方面发挥了重要作用,寻找特异性遗传标记鉴定人体不同生物性物质来源是近年来法医学工作者研究的重点和难点。本文就目前研究较多的用于人体生物性物质来源鉴定的遗传标记进行综述,包括DNA甲基化、mRNA、microRNA、微生物菌群、蛋白质等。通过比较不同种类遗传标记鉴定人体生物性物质来源的原理和方法,发现人体不同生物性物质来源的鉴定都有其最适合的遗传标记种类,并且可以采用单一遗传标记或联合多种遗传标记进行检验。尽管目前各法医学实验室无统一的标准和方法鉴定人体生物性物质来源,但研究开发一系列成熟可靠的方法区分人体不同生物性物质,进而发挥其法庭证据作用,将是未来的发展方向。     

Forensic drug intelligence: an important tool in law enforcement

Organised criminality is a great concern for national/international security. The demonstration of complex crimes is increasingly dependant on knowledge distributed within law-enforcement agencies and scientific disciplines. This separation of knowledge creates difficulties in reconstructing and prosecuting such crimes. Basic interdisciplinary research in drug intelligence combined with crime analysis, forensic intelligence, and traditional law enforcement investigation is leading to important advances in crime investigation support. Laboratory results constitute one highly dependable source of information that is both reliable and testable. Their operational use can support investigation and even provide undetected connections or organisation of structure. The foremost difficulties encountered by drug analysts are not principally of a chemical or analytical nature, but methodologies to extract parameters or features that are deemed to be crucial for handling and contextualising drug profiling data. An organised memory has been developed in order to provide accurate, timely, useful and meaningful information for linking spatially and temporally distinct events on a national and international level (including cross-border phenomena). Literature has already pointed out that forensic case data are amenable for use in an intelligence perspective if data and knowledge of specialised actors are appropriately organised, shared and processed. As a particular form of forensic case data, the authors' research focuses on parameters obtained through the systematic physical and chemical profiling of samples of illicit drugs. The procedure is used to infer and characterise links between samples that originate from the same and different seizures. The discussion will not, however, focus on how samples are actually analysed and compared as substantial literature on this topic already exists. Rather, attention is primarily drawn to an active and close collaboration between magistrates, forensic scientists, law enforcement investigators and crime analysts from different institutions with the aim of generating, using and validating relevant profiling case data as integral part of investigative and crime analysis processes. Original advances are highlighted through experiences from criminal investigations of offences related to the unlawful importation, exportation, supply and possession of illicit drugs.     

Application of the BioMek 2000 Laboratory Automation Workstation and the DNA IQ System to the extraction of forensic casework samples

Robotic systems are commonly utilized for the extraction of database samples. However, the application of robotic extraction to forensic casework samples is a more daunting task. Such a system must be versatile enough to accommodate a wide range of samples that may contain greatly varying amounts of DNA, but it must also pose no more risk of contamination than the manual DNA extraction methods. This study demonstrates that the BioMek 2000 Laboratory Automation Workstation, used in combination with the DNA IQ System, is versatile enough to accommodate the wide range of samples typically encountered by a crime laboratory. The use of a silica coated paramagnetic resin, as with the DNA IQ System, facilitates the adaptation of an open well, hands off, robotic system to the extraction of casework samples since no filtration or centrifugation steps are needed. Moreover, the DNA remains tightly coupled to the silica coated paramagnetic resin for the entire process until the elution step. A short pre-extraction incubation step is necessary prior to loading samples onto the robot and it is at this step that most modifications are made to accommodate the different sample types and substrates commonly encountered with forensic evidentiary samples. Sexual assault (mixed stain) samples, cigarette butts, blood stains, buccal swabs, and various tissue samples were successfully extracted with the BioMek 2000 Laboratory Automation Workstation and the DNA IQ System, with no evidence of contamination throughout the extensive validation studies reported here.     

1.1. HLA and other leucocyte and platelet alloantigens in paternity testing (introductory lecture)

The analysis of lipstick stains presented here is a combination of several techniques. In addition to colour comparison and thin-layer chromatographic (TLC) analysis, X-ray analysis in a scanning electron microscope and high-performance liquid-chromatographic (HPLC) analysis have been used. X-ray analysis may be performed directly on the material bearing the stain. For TLC and HPLC analyses a solvent extract of the stain was used.Guidance is given for practical examination of lipstick stains. When using all the methods described in this paper, all the 117 different lipstick samples examined in this study could be distinguished from each other.     

Humans Are Animals,Too: Critical Commonalities and Differences Between Human and Wildlife Forensic Genetics

Wildlife forensics has recently been recognized among the wide variety of forensic science disciplines. This review compares human and wildlife DNA forensics, which use the same genetic tools, but often for far different purposes. Human forensic genetics almost invariably attempts to identify individual perpetrators involved in a given crime. Wildlife forensics often determines whether a crime has occurred. In addition to techniques familiar in human laboratories, like individual matching with STRs, wildlife analysts may be asked to determine the taxonomic identity, geographic source, or sex of evidence items, or the familial relationships or minimum number of individuals among a group of samples. This review highlights the common questions, legal framework, databases, and similar validation requirements to foster understanding between disciplines. Based on this understanding, human and wildlife DNA practitioners may work together and learn from each other in order to elevate the discipline of forensic genetics.