Forensic discrimination of menstrual blood and peripheral blood using attenuated total reflectance (ATR)-Fourier transform infrared (FT-IR) spectroscopy and chemometrics.

Body fluids are one of the most important pieces of evidence encountered in forensic cases especially in cases of sexual assault. Analysis of such evidence can help to establish a link between the perpetrator, the victim, and the crime scene and thereby assist in crime reconstruction. However, one of the biggest challenges faced by the investigators in sexual assault cases is that of ascertaining the issue of consent of the victim. In this matter, differentiation of menstrual blood (either in dried or stained form) from traumatic peripheral blood can give a potential solution on this particular aspect. A number of studies have been attempted to differentiate these two body fluids using various biochemical and serological methods. However, the methods employed are limited by factors such as sample destructivity and non-specificity, and the methods are susceptible to false positive results. In the present study, the scope of attenuated total reflectance (ATR)-Fourier transform infrared (FT-IR) spectroscopy in discriminating samples of menstrual blood and peripheral blood has been investigated, in combination with chemometric tools such as principal component analysis (PCA), partial least square regression (PLSR), and linear discriminant analysis (LDA). PCA resulted in 93.3% accuracy, whereas PLSR and LDA resulted in 100% accuracy for the discrimination of peripheral blood from menstrual blood. Application of PCA for the discrimination of menstrual blood from vaginal fluid and seminal fluid delivered 100% classification. Similarly, 100% classification was achieved while differentiating between menstrual blood and blood look-alike substances. Furthermore, in the current study, the effect of substrates on the analysis of menstrual blood has also been studied and described. Graphical Abstract.

Discrimination of electrical tapes using ATR-FTIR spectroscopy and chemometrics.

Electrical tapes are recovered during criminal investigations as physical evidence in cases of rape, kidnapping, and explosion incidents. The analysis of such evidence can provide an evidentiary link between the suspect, the victim, object, or the crime scene. In the present study, 25 brands of electrical tapes have been analyzed using ATR-FTIR (attenuated total reflectance Fourier transform infrared) spectroscopy. Samples (1 cm2 ) were analyzed in the mid IR (Infrared) region from 4000-600 cm-1 , and the functional groups of various components have been profiled. Chemometric methods-PCA (principal component analysis) and PCA-LDA (linear discriminant analysis) have been employed to interpret the data and classify the samples into its respective classes. Preliminary assessment of sample clustering due to similar chemical composition was visualized using PCA. PCA-LDA applied for classification purpose yielded classification accuracy (calibration) of 92.98% for the adhesive side and 88% for the backing side. The validation results showed classification accuracy of 89.47% for the adhesive side and 84% for the backing side. Blind validation study was carried out using 5 samples, and classification accuracy of 100% and 80% was obtained for the adhesive and the backing side, respectively. In the current study, a preliminary substrate study was carried out, and the results showed that the backing samples could be more accurately matched to their correct source of origin than the adhesive side.

Chemometric analysis on ATR-FT-IR spectra of spray paint samples for forensic purposes.

Forensic investigation of cases involving spray paints requires comparison of paint such as those recovered from graffiti with the references sample seized from the suspect's possession site. This study describes the use of Attenuated Total Reflectance Fourier Transform Infrared (ATR-FT-IR) spectroscopy for nondestructive analysis of 20 red spray paints of different manufactures, which could be encountered at a crime scene, particularly in case of vandalism. Spray paints could be encountered on a number of different substrates, and therefore, the samples were placed on simulated substrates and were analyzed in order to check the effect of substrates on sample analysis. Results showed that all samples could be successfully differentiated using ATR-FT-IR spectroscopy assisted with principal component analysis. Analysis of spray paints on a paper and fabric substrate resulted in poor spectra and hampered the comparative examination between the spray paint deposited on fabric/paper surface and the control sample. In case of sample deposited on cemented wall, the substrate got scraped out along with the sample and caused interferences in the spectrum. On the other hand, for sample deposited on the floor, gloves, metal, plastic, leather shoes, tile, wood, and hair substrates, all the significant peaks were observed to be present and hence it was concluded that comparative studies can be done between neat spray paint and spray paint deposited on the aforesaid substrates. Additionally, blind validation test was conducted and it resulted in linking the blind samples with the source or origin with 100% accuracy.

Discrimination of vermilion (sindoor) using attenuated total reflectance fourier transform infrared spectroscopy in combination with PCA and PCA-LDA.

Vermilion (sindoor) is considered sacred in the Hindu religion, and it is used routinely throughout the world by married Hindu women along the line of hair parting during marriage ceremonies, religious rituals, and festivals. Owing to its esthetic appeal, it is sometimes illegally used as a food additive; leading to potential health risks. Therefore, due to the aforementioned reasons, vermilion can likely be encountered as trace evidentiary material during crime investigations, particularly in cases of sexual and physical offenses against women. Analysis of such evidence can provide a link between the criminal, the victim, and the crime scene and thereby be utilized as associative evidence in the court of law. In the present study, ATR-FTIR spectroscopy has been used for the examination of 37 different manufacturers of vermilion. Chemometric methods such as principal component analysis (PCA) and PCA-LDA were performed on the obtained spectra for objective interpretation of results. PCA delivered 99.06% discrimination of samples while PCA-LDA employed for classification purpose delivered 95.25% calibration accuracy and 88% validation accuracy. Afterward, the validity of the chemometric methods employed was tested by blind testing of samples. A preliminary study on the effect of selected substrates (cotton cloth, tissue paper, glass, and plastic) on sample analysis indicates that while sample stain on substrates could be linked to its parent source even after a month, linking an aged samples (after 8 months) could be hindered due to evaporation of components present in vermilion. Overall, the current methodology utilized has a potential prospect in future forensic-casework.

Detection of bloodstains using attenuated total reflectance-Fourier transform infrared spectroscopy supported with PCA and PCA-LDA.

The most important task in a criminal investigation is to detect and identify the recovered biological stains beyond reasonable scientific doubt and preserve the sample for further DNA analysis. In the light of this fact, many presumptive and confirmatory tests are routinely employed in the forensic laboratories to determine the type of body fluid. However, the currently used techniques are specific to one type of body fluid and hence it cannot be utilized to differentiate multiple body fluids. Moreover, these tests consume the samples in due process, and thus it becomes a great limitation especially considering the fact that samples are recovered in minute quantity in forensic cases. Therefore, such limitations necessitate the use of non-destructive techniques that can be applied simultaneously to all types of bodily fluids and allow sample preservation for further analysis. In the current work, attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy has been used to circumvent the aforementioned limitations. The important factors which could influence the detection of blood such as the effect of substrates, washing/chemical treatment, ageing, and dilution limits on the analysis of blood have been analysed. In addition, blood discrimination from non-blood substance (biological and non-biological in nature) has also been studied. Chemometric technique that is PCA-LDA has been used to discriminate blood from other body fluids and it resulted in 100% accurate classification. Furthermore, blood and non-blood substances including fake blood have also been classified into separate clusters with a 100% accuracy, sensitivity, and specificity. All-inclusive, this preliminary study substantiates the potential application of ATR-FTIR spectroscopy for the non-destructive identification of blood traces in simulated forensic casework conditions with 0% rate of false classification.

Rapid and non-destructive analysis of eye-cosmetics using ATR-FTIR spectroscopy and chemometrics.

Cosmetic evidence recovered during crime investigations, particularly in cases of physical and sexual assault against women can be utilised as associative evidence in the court of law. This evidence can provide a link between the suspect, the victim, and the crime scene and assist in solving criminal cases. A mismatched profile of exhibit's source of origin can also be utilised to definitely exclude the suspect exhibits. In the present study, ATR-FTIR (attenuated total reflectance-fourier transform infrared) spectroscopy has been employed for the analysis of eye-cosmetics (eyeliner and eyeshadow) samples. Chemometric tool- PCA (principal component analysis) has been used for the recognition of patterns in the data. PCA-LDA (linear discriminant analysis) utilized for classification purpose showed calibration accuracy of 100% and 98% for eyeliner and eyeshadow respectively while validation result showed 97% and 97% respectively. Preliminary substrate study has been performed in the current study. Result suggests that substrates such as cotton cloth and tissue paper hinder the analysis of eyeliner while the stain of eyeshadow on substrates such as cotton cloth, tissue paper, glass, and plastic could be correctly matched with its parent source.

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.

Forensic entomotoxicology: Current concepts, trends and challenges.

The application of forensic entomotoxicology in investigations of death by poisoning has gained popularity as insects serves as an alternative specimen (evidence) when conventional toxicological samples have degraded or are no longer available. Successful detection, identification, and quantification of drugs and toxic substances from insects have been demonstrated through various research experiments. In the present review paper, role of insects as toxicological sample, its collection and preservation, analysis using various instrumental techniques, and trends in the use of analytical techniques have been discussed. Limitations hindering the growth of this field and the way forward for future studies have been highlighted. In addition, the effect of poisons on insects used for postmortem interval estimation has been described.

Differentiation of locally manufactured Kajal by Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy supported by chemometric analysis.

Kajal, also known as Kohl (Arabic name) a cosmetic, is used for the beautificationof eyes and face as a whole. It can serve as an important corroborative evidence in various crimes against women and can also be used as a writing instrument to write the anonymous letters, threatening notes, suicide letters on a variety of substrates such as cotton cloth, tissue paper, tissue wipes, skin etc. 40 Kajal samples from 33 different brands were collected and analyzed using Attenuated Total Reflectance Fourier Transform Infrared (ATR FT-IR) Spectroscopy with advance statistical and computational tools. The present research investigated the application of this technique for the differentiation of locally manufactured Kajal based on the differences in the peak patterns, frequencies, and strengths of peak exhibited within the fingerprint region. The blind test was conducted with four unknown samples which were pretended to be unknown to the analyst and it was observed that all the unknown samples were predicted correctly with their respective classes. Effect of substrates and ageing were also studied under similar set of experimental conditions. This technique has been proven to be of high significance with complete non-destructive nature requiring minimal or no sample preparation and high repeatability.