Forensic analysis of biological fluid stains on substrates by spectroscopic approaches and chemometrics: A review

Bodily fluid stains are one of the most relevant evidence that can be found at the crime scene as it provides a wealth of information to the investigators. They help to report on the individuals involved in the crime, to check alibis, or to determine the type of crime that has been committed. They appear as stains in different types of substrates, some of them porous, which can interfere in the analysis. The spectroscopy techniques combined with chemometrics are showing increasing potential for their use in the analysis of such samples due to them being fast, sensitive, and non-destructive.This is a comprehensive review of the studies that used different spectroscopic techniques followed by chemometrics for analysing biological fluid stains on several surfaces, and under various conditions. It focuses on the bodily fluid stains and the most suitable spectroscopic techniques to study forensic scientific problems such as the substrate's characteristics, the influence of ambient conditions, the aging process of the bodily fluids, the presence of animal bodily fluids and non-biological fluids (interfering substances), and the bodily fluid mixtures. The most widely used techniques were Raman spectroscopy and attenuated total reflection Fourier transform infrared spectroscopy (ATR FTIR). Nonetheless, other non-destructive techniques have been also used, like near infrared hyperspectral imaging (HSI-NIR) or surface enhanced Raman spectroscopy (SERS), among others. This work provides the criteria for the selection of the most promising non-destructive techniques for the effective in situ detection of biological fluid stains at crime scene investigations.The use of the proper spectroscopic and chemometric approaches on the crime scene is expected to improve the support of forensic sciences to criminal investigations. Evidence may be analysed in a non-destructive manner and kept intact for further analysis. They will also speed up forensic investigations by allowing the selection of relevant samples from occupational ones.