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.     

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.     

Investigation of the new possibility of mathematical processing of Raman spectra for dating documents

The use of spectral analysis methods to determine the age of writing inks is an important forensic task. However, the use of spectral data for this purpose has a number of limitations and difficulties. This paper considers the application of the Raman spectroscopy method to an urgent forensic task. The known mechanisms of dye degradation are analyzed; Raman bands are identified that are related to the age of the sample. In a sample of 5 randomly selected writing inks, temporary markers were identified. Narrow sections of Raman spectra containing characteristic lines were used for analysis. It was shown that processing narrow sections of the Raman spectra using the PCA chemometric method allowed the separation of writing inks into groups (clusters) corresponding to different creation intervals.     

Diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) detection limits for blood on fabric: Orientation and coating uniformity effects

Diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) was used to analyze four types of forensically relevant fabrics coated with varying dilutions of blood. The blood was applied in two manners, dip coating with a smooth and uniform layer and drip coating with droplets from pipettes. Spectra of neat and dip coated fabrics were acquired using controlled orientations, and these were compared to spectra collected on samples with random orientations. The improved reproducibility seen in visual inspection of the spectra is confirmed by principal component and linear discriminant projections of the spectra, as well as by statistical hypothesis testing. Principal component regression (PCR), using the regions of the IR spectra associated with the amide A/B, I, II, and III vibrational bands (3500–2800, 1650, 1540, and 1350 cm⁻¹), was employed on the more uniform dip coated spectra to estimate limits of detection for blood on two of the four fabrics – acrylic and nylon. These results demonstrate that detection limits for blood on fabrics can be decreased significantly by controlling for the orientation and face of the fabric samples while collecting spectra. Limits of detection for acrylic and nylon were found to be 196 × and 227 × diluted blood, respectively.     

Identification and isolation of male cells using fluorescence in situ hybridisation and laser microdissection, for use in the investigation of sexual assault

In cases of sexual assault involving an azoospermic assailant, vaginal swabs taken from the victim may fail to provide an autosomal DNA profile with which to search a suspect database, as the signal from any male cells present would be masked by that from the overwhelming number of female cells collected on the swab. Here, we describe a method of visually identifying diploid male cells in such samples using fluorescence in situ hybridisation, and selectively harvesting them by means of laser microdissection. This combination of techniques was tested on 26 post-coital vaginal swabs taken at a range of times after intercourse; the collected cells were then subjected to a simple lysis procedure and DNA was amplified using the AmpFlSTR^® SGMPlus^® multiplex under low copy number conditions. Useful DNA profiles were generated from samples taken up to 24 h after intercourse.