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.