Application of Infrared and Raman Spectroscopy in Paint Trace Examination

Raman spectroscopy has proved to be a promising technique in forensic examinations, where optical microscopy, micro‐infrared spectroscopy, and microspectrophotometry in the visible and UV range are used for identification and differentiation between paint traces. Often no organic pigments are detected using Fourier transform infrared spectroscopy, because their content in a trace is usually undetectable. Application of a micro‐Raman spectrometer equipped with several excitation lasers helps forensic experts in paint analysis enabling identification of main pigments. Three cases concerning comparative examination of car paint are discussed in detail. The comparison of Raman spectra of paint chips found on clothing of a victim or smears found on body of a damaged car to these of paint chips originated from the suspected car enabled us to identify the car involved in the accident. When no comparative material is available, the method can be useful in establishing the color and make of the car.     

Characterization of Blue Pigments Used in Automotive Paints by Raman Spectroscopy

Micro‐Raman spectroscopy was applied to forensic identification of pigments in paint chips and provided differentiation between paint samples. Sixty‐six blue automotive paint samples, 26 solid and 40 metallic were examined. It was found that the majority of the collected Raman spectra provided information about the pigments present. However, in some cases, fluorescence precluded pigment identification. Using laser excitation at longer wavelengths or pretreatment to effect photobleaching often resulted in reduced fluorescence, particularly for solid color samples, and allowed pigment identification. The examined samples were compared pairwise taking into account number, location, and intensity of absorption bands in their infrared spectra. The estimated discrimination power ranged from 97% for solid paint samples to 99% for metallic paint samples.     

红外差谱技术用于微量混合油漆的比对检验

对交通事故中的漆片检材进行同一物质比对时,某些情况下难以提取到单一的油漆碎片而是两辆车的油漆混合在一起,给物证比对带来困难。红外差谱技术具有辨别混合物中不同成分的功能,因此可应用于交通事故中混合油漆检材的比对检验。本文对一起交通事故的漆片检材,分别用三种方式提取、制样,用红外光谱仪进行分析,借助红外差谱技术对谱图进行处理,排除混合油漆的干扰。处理后得到的谱图与样本的谱图进行比对,结果显示两者主要官能团吸收峰位置、峰型、峰强度基本一致,可出具比对一致的结论。针对混合油漆检材的比对检验,红外差谱技术可提供支持。     

The Evidentiary Significance of Automotive Paint from the Northeast: A Study of Red Paint

This population study was conducted to assess the frequency of physical, microscopical, and chemical properties of automotive paint chips. Population studies of trace evidence provide valuable analytical data for criminalists to assess evidentiary significance. Two‐hundred automotive paint chips were collected from auto body shops from the Northeastern United States. All samples were analyzed using stereomicroscopy, brightfield, and polarized light microscopy. Red paints were targeted for further analysis using a sequence of modern instrumental techniques commonly used by forensic paint examiners: Fourier‐transform infrared (FT‐IR), Raman, and ultraviolet–visible (UV–Vis) microspectroscopy. The discrimination potential of each analytical method was evaluated by inter‐comparing the paint samples. Results demonstrated that macroscopic and microscopic properties were able to differentiate 99.995% of the population (one undifferentiated pair out of 19,900). When combined with either FT‐IR or UV–Vis microspectroscopy, all paints were differentiated. The results of this research lead to the conclusion that one would not expect to encounter two indistinguishable paint chips originating from different sources during the investigation of a single event.     

Forensic applications of infrared chemical imaging: multi-layered paint chips

This paper examines the potential of infrared chemical (hyperspectral) imaging as a technique for the forensic analysis of automotive paint chips in particular, and multicomponent (e.g., layered) samples in general. Improved sample preparation procedures for the infrared analysis of paint chips are detailed, with the recommendation that where mounting resins are chemically incompatible with the sample, it is better to mount and section the sample in a soft wax from which the sections can be removed and pressed into a KBr disk for transmission analysis. Infrared chemical images of multilayered paint chips have been successfully obtained, with the chief advantage over conventional infrared analysis being that thousands of infrared spectra are collected in a few minutes across the whole sample, at a spatial resolution of around 5 microm. As with conventional infrared spectroscopy, chemical species can be identified from their spectra, but the wealth of information available can be also extracted in a number of different ways that make multicomponent spectral (and hence chemical) comparisons between two samples easy to visualize and understand. In one approach, the infrared chemical images of two paint chips being compared side-by-side can be viewed as a "movie," in which each frame is an intensity map of the two samples at a given wavenumber (frequency) value. In another approach, the spectra (pixels) in the image files are classified into chemically similar groups, resulting in a "cluster" image that makes it possible to simultaneously compare all of the layers in two paint chips. These methods are applicable to other multicomponent samples, and also to other chemical imaging techniques.     

显微光谱成像技术在汽车油漆截面检验中的应用

目的对汽车多层油漆的横截面进行分层和比对研究。方法运用显微光谱成像技术对收集的12个省市623个汽车漆片样本进行分层,并对截面结构相同或相似的油漆样本进行了区分。结果 623个油漆样本均可以分层,分析出漆层为两层及以上的油漆样本数占总数的87.26%;截面结构相同或相似的油漆样本能够多层同时区分。结论该方法具有科学及实用价值,检验结果可靠,可见光谱成像技术完全可运用拓展到微量物证的检验中。     

Elemental analysis of glass fragments by ICP-MS as evidence of association: analysis of a case

Inductively coupled plasma mass spectrometry (ICP-MS) was used to analyze glass evidence from a case in which a person broke a window in each of 15 vehicles in a parking lot in order to gain entry into the vehicles. The results of the analysis by traditional methods, which measure the properties of color, thickness, density, and refractive index, are also reported. A total of 15 known samples representing the windows on the cars and 42 questioned glass fragments recovered from the suspect and the police vehicle where the suspect was sitting were submitted for analysis. Density comparisons separated one of the known samples into three samples, increasing the number of known samples from 15 to 17. The concentrations of 16 elements were measured for all but three of the samples using an external calibration ICP-MS method with internal standardization. Color assessment (non-instrumental) separated the 17 known samples into two groups, and refractive index measurements resulted in six groups when the Emmons double variation method was used and ten groups when the Glass Refractive Index Measurement 2 (GRIM2) system was used. Elemental analysis, by itself, differentiated all of the known samples from each other and associated four of the known sample fragments with several of the questioned sample fragments. The informing power of RI, density, and elemental analysis comparisons is evaluated and a summary of the case results is reported.     

Applications of Blue Light‐curing Acrylic Resin to Forensic Sample Preparation and Microtomy

This study discusses the results of an evaluation of a one‐part blue light‐curing acrylic resin for embedding trace evidence prior to the preparation of thin sections with a microtome. Through a comparison to several epoxy resins, the physical properties relevant to both trace evidence examination and analytical microscopy in general, including as viscosity, clarity, color, hardness, and cure speed, were explored. Finally, thin sections from paint samples embedded in this acrylic resin were evaluated to determine if, through smearing or impregnation, the resin contributed to the infrared spectra. The results of this study show that blue light‐curing acrylic resins provide the desired properties of an embedding medium, generate high‐quality thin sections, and can significantly simplify the preparation of paint chips, fibers and a multitude of other types of microscopic samples in the forensic trace evidence laboratory.     

Use of RGB values in the Periodic Acid-Schiff color test to determine the presence of vaginal fluid

This study demonstrates how RGB color values from microscopic smears stained with the Periodic Acid-Schiff reagent under standardized microscopy conditions can be used to indicate the presence of vaginal secretions. Based on data obtained in the study, a numeric threshold determined from the sum of separate values for red, blue and green was determined to differentiate vaginal-based samples with other body fluids. Using this threshold, 55 of 57 vaginal-based samples tested positive for the presence of vaginal secretion. Conversely, 27 of 29 smears prepared from other body fluids yielded negative results. However, when graphing RGB sum values against a calculated RGB integer no overlap in data was obtained between all vaginal-based samples and other body fluid samples, clearly differentiating them. One-way ANOVA testing with a 95% confidence interval indicated that vaginal samples from different age groups showed no difference in RGB sum values. Similarly, the location that vaginal swabs were collected (from the outside of a condom or a vaginal swab) also showed no statistical difference using one-way ANOVA at 95% confidence. Furthermore, refrigerated test swabs aged up to 15 months showed no demonstrable differences. Pair-wise t-testing using RGB sum values, however, did show significant differences between vaginal samples and all other body fluids tested. Finally, the method successfully differentiated between pre-and post-coital penile swabs and finger swabs taken before and after digital vaginal penetration in anecdotal comparisons using the method.     

Locating,Identifying, and Comparing Sub‐visible Paint Particles

Forensic paint comparisons are generally conducted on samples which, while small relative to their source, are still visible to the unaided eye and are thus located and analyzed without great difficulty. Here we demonstrate that a more detailed examination of candidate transfer surfaces can capture materials (questioned samples), even when such traces are invisible to the unaided eye. While certain analytical details (such as layer sequence or a pure FTIR spectrum) may not be obtainable from such traces due to their size and condition, a detailed analysis of the sample characteristics that are analytically accessible may still provide sufficient analytical data to arrive at a probative result. Here we present the application of this approach to a suspected paint transfer case, involving particles of paint as small as 40 μm in size. Using a combination of stereomicroscopy, polarized light microscopy, infrared microspectroscopy, Raman microspectroscopy, and SEM/EDS, all performed on a single, subsample of the original minute particle, it was possible to demonstrate evidence of a two‐way transfer between the suspected sources. Furthermore, the transferred paint particle in one direction could be classified as automotive in nature based on a combination of polymer composition, microscopic texture, and pigment package (which included three specifically identified pigments). This work demonstrates (i) the potential for improving detection limits when searching for a questioned sample, (ii) the potential benefits of higher resolution analyses on samples that would be traditionally labeled as “sample‐size limited,” and (iii) the value of case‐specific interpretation over standardized, one‐size fits all report templates.     

LC—MS／MS法测定蓝色圆珠笔油墨中的碱性染料

目的建立蓝色圆珠笔油墨中碱性染料的LC—MS／MS方法,为蓝色圆珠笔油墨的种类鉴别提供方法。方法用二级质谱寻找并确定结晶紫、甲基紫、维多利亚蓝B、碱性紫14、碱性蓝7和罗丹明B等碱性染料的特征性母离子／子离子对。收集50种蓝色圆珠笔．划线后对其笔道用0．5mm直径打孔器取样,乙腈超声提取。液相分离采用WatersXBridgeC18柱。流动相为0．1％甲酸缓冲液（A）-乙腈（B）,梯度程序洗脱。结果4个点的取样量足以满足检测需要,采用相对峰面积的定量方法．结果重现性好．RSD％≤2．3％。应用该方法对50种蓝色圆珠笔油墨中的碱性染料进行检测,区分率为94．4％。结论所建LC—MS／MS方法定性准确,定量可靠,为蓝色圆珠笔油墨的种类鉴别提供了方法。     

Microscopical discrimination of twins' head hair

Twin populations are ideal for studying human variation; a study of twin's hair, therefore, provided a better understanding of the value of hair comparisons. Duplicate head hair samples from 17 pairs of twins and one set of identical triplets were compared in a verified blind study. In addition to the direct comparison of all twins, random samples of two or three hairs were compared with randomly selected groups of known samples in a second blind study, to better simulate an ordinary forensic science case. Features commonly used by forensic hair examiners were adequate to distinguish hair samples from each twin from all other samples, illustrating the power of microscopical comparison when numerous questioned hairs are available in evidence. When two or three hairs were compared with randomly selected known samples, several were indistinguishable from hair samples other than the true source, proving once again that a human hair can never be associated with one person to the exclusion of all others.     

常见油漆稀料的GC／MS的检验及分类研究

目的对常用油漆稀料的检验分析方法及分类进行研究,并建立SPME-GC／MS对纵火案件中油漆稀料快速、灵敏、准确的分析方法。方法采用SPME-GC／MS法对5种不同型号品牌的常见油漆稀料进行分析。结果确定了样品处理方法和仪器分析最佳条件。实验的5种油漆稀料样本根据组分特征和结构特征的不同可归纳为4类。结论该研究结果可为纵火案件中油漆稀料残留物提供一种新的检验方法和检验依据。     

SEM检验漆膜厚度鉴别同种颜色的汽车油漆

目的研究扫描电子显微镜测量漆膜厚度的方法对颜色相近的汽车油漆的区分能力。方法应用扫描电子显微镜对不同汽车厂家生产的12块蓝色油漆进行总厚度及分层厚度测量,并对所得数据进行t检验分析。结果在所有两两比对的66组样品对中,在置信度为95％时,用总厚度及分层厚度进行区分,区分率达到100％。结论用扫描电子显微镜微观测量油漆样品总厚度及分层厚度的方法,可对颜色相近的汽车油漆进行有效区分。     

Recovery of Spray Paint Traces from Clothing by Beating

Manual recovery of spray paints from textiles using a microscope, the routine method in many laboratories, is often laborious. Beating the clothing with a plastic rod, the routine method used for recovery of glass traces within the authors’ laboratory, is proposed as an alternative. The efficiency of the method was evaluated by spray tests with fluorescent paint. In these tests, paint particles in the acquired debris samples, as well as those remaining on the textiles, were investigated. The results show that beating is an efficient way to recover and concentrate paint particles. A good efficiency for jeans fabric and rough knitwear is reported. The results appeared to be less satisfactory for smooth woven fabric. Application of the method in casework was effective for graffiti paints as well as for flaked car paint.     

A novel histological technique for distinguishing between epithelial cells in forensic casework

There are a number of forensic cases in which the identification of the epithelial cell type from which DNA originated would provide important probative evidence. This study aimed to develop a technique using histological staining of fixed cells to distinguish between skin, buccal and vaginal epithelium. First, 11 different stains were screened on formalin-fixed, wax-embedded cells from five women. Samples were analysed qualitatively by examining staining patterns (colour) and morphology (absence or presence of nuclei). Three of the staining methods – Dane's, Csaba's and Ayoub-Shklar – were successful in distinguishing skin epithelial cells from buccal and vaginal. Second, cells were smeared directly onto slides, fixed with one of five fixatives and stained with one of the three stains mentioned above. Methanol fixation, coupled with the Dane's staining method, specific to keratin, was the only technique that distinguished between all three cell types. Skin cells stained magenta, red and orange and lacked nuclei; buccal cells stained predominantly orange–pink with red nuclei; while vaginal cells stained bright orange with orange nuclei and a blue extracellular hue. This staining pattern in vaginal cells was consistent in samples collected from 50 women aged between 18 and 67. Identification of cell type from unlabelled micrographs by 10 trained observers showed a mean success rate of 95%. The results of this study demonstrate that histological staining may provide forensic scientists with a technique for distinguishing between skin, buccal and vaginal epithelial cells and thus would enable more conclusive analyses when investigating sexual assault cases.     

汽车罩光漆的ATR-FTIR法检验

目的通过对油漆样品罩光漆成膜物质的检验以区分不同汽车厂家的油漆样品。方法采用衰减全反射红外光谱法（Fourier Transform Infrared Spectroscopy Attenuated Total Reflection Spectroscopy,ATR-FTIR）研究15个汽车厂家提供的356份油漆样品的罩光漆成膜物质。结果在所有油漆样品中,丙烯酸漆出现的频率最高,氨基漆出现的频率最低;在所有厂家中,使用丙烯酸漆的厂家最多,使用氨基氨酯混合漆的厂家最少;在使用相同罩光漆的不同厂家的油漆样品中,通过比较红外谱图可以对样品进一步区分。结论 ATR-FTIR法能够在一定程度上区分不同汽车厂家的油漆样品,从而大大缩小侦查范围,对于涉车案件的快速侦破具有非常重要的作用。     

Identification of colorants in pigmented pen inks by laser desorption mass spectrometry

Pigments are rapidly replacing dyes as colorants in pen and printer inks, due to their superior colors and stability. Unfortunately, tools commonly used in questioned document examination for analyzing pen inks, such as TLC, cannot be used for the analysis of insoluble pigments on paper. Laser desorption mass spectrometry is demonstrated here as a tool for analyzing pigment-based pen inks. A pulsed nitrogen laser can be focused onto a pen stroke from a pigmented ink pen on paper, and positive and negative ions representative of the pigment can be generated for subsequent mass spectrometric analysis. Targeted pens for this work were a set of Uni-ball 207 pigmented ink pens containing blue, light blue, orange, green, violet, red, pink, and black inks. Copper phthalocyanine was identified as the pigment used to make both blue inks. A mixture of halogenated copper phthalocyanines were identified in the green ink. Unexpectedly, the pink ink was found to contain a red pigment, Pigment Red 12, treated with a mixture of water-soluble dyes. Each sample yielded ions representative of the pigments present.     

Application of the micro-FTIR spectroscopy, Raman spectroscopy and XRF method examination of inks

In routine examination of inks on questioned documents non-destructive analytical methods, such as microscopic and optical techniques are applied first. However, they are often insufficient to identify the inks used for the preparation of the document. In such cases, it is necessary to apply chemical methods that normally cause partial destruction of the examined material. The aim of this work was to evaluate the possibility of discrimination between inks by the use of spectrometric methods, i.e. micro-FTIR spectroscopy, Raman spectroscopy and XRF. About 70 samples of blue and black ballpoint pen and gel inks were examined. It was found that about 90% of the samples of the same type and colour could be distinguished using these methods.     

SEM-EDS analysis and discrimination of forensic soil

Soils vary among different areas, and have some characteristics because of the natural effects and transfers made by human and other living beings in time. So that forensic examination of soil is not only concerned with the analysis of naturally occurring rocks, minerals, vegetation, and animal matter. It also includes the detection of such manufactured materials such as ions from synthetic fertilizers and from different environments (e.g., nitrate, phosphate, and sulfate) as environmental artifacts (e.g., lead or objects as glass, paint chips, asphalt, brick fragments, and cinders) whose presence may impart soil with characteristics that will make it unique to a particular location. Many screening and analytical methods have been applied for determining the characteristics which differentiate and discriminate the forensic soil samples but none of them easily standardized. Some of the methods that applied in forensic laboratories in forensic soil discrimination are the color comparison of the normal air-dried (dehumidified) and overheated soil samples, macroscopic observation, and low-power stereo-microscopic observation, determination of anionic composition by capillary electrophoresis (CE), and the elemental composition by scanning electron microscope (SEM)-energy dispersive X-ray spectrometer (EDS) and other high sensitivity techniques. The objective of this study was to show the effect of the application of 9 tonnes/cm2 pressure on the elemental compositions obtained by SEM-EDS technique and comparing the discrimination power of the pressed-homogenized and not homogenized forensic soil samples. For this purpose soil samples from 17 different locations of Istanbul were collected. Aliquots of the well mixed samples were dried in an oven at 110-120 degrees C and sieved by using 0.5 mm sieve and then the undersieve fraction(<0.5 mm) of these samples put on an adhesive tape placed on a stub. About 100-150 mg aliquots of dried, sieved samples were pressed under 9 tonnes/cm2 pressure by KBr disk preparation apparatus of an infrared spectrophotometer. Surfaces of the randomized particles and the pressed disks of the soil samples were scanned and the elemental compositions were determined with scanning electron microscope JEO-JSM-5600 equipped with an energy dispersive X-ray spectrometer OXFORD Link-ISIS-300. The samples from top of the sieves were examined with stereo-microscope equipped with JVC-TK-128DE color video camera and JVC-GV-PT2, digital video printer. Natural and artificial materials that have characteristic features were identified. Then for additional confirmation all soil samples were dried at 120 degrees C and over 780 degrees C and their colors compared. We concluded that pressing the whole sieved soil samples under 9 tonnes/cm2 pressure results in smashing over the harder particles into the softer matrix and results in homogenization of the soil sample. The elemental compositions of these samples obtained by SEM-EDS with 10-fold less standard deviation (S.D.) values and so that with more reproducibility and discrimination power.