Detection of Oral Fluid Stains on Common Substrates Using SEM and ATR–FTIR Spectroscopy for Forensic Purposes

Attenuated total reflectance (ATR) Fourier-transform infrared (FTIR) spectroscopy has been pursued as a novel approach to detect and differentiate biological materials with high specificity owing to its ability to record unique spectral patterns corresponding to the biochemical composition of a specimen. This study expands the application of ATR-FTIR for detecting oral fluid (OF) stains on various common substrates, including four porous and six nonporous substrates. For nonporous substrates, the spectral contribution from the substrate was minimal, and no background subtraction from the substrate bands was required (except for mirrors). For porous substrates, the contribution from the surface was pronounced and was addressed via background subtraction. The results indicated that major OF bands were detected on all the surfaces, even six months after OF deposition. Furthermore, scanning electron microscopy (SEM) was used to probe the morphologies of OF stains on various substrates. SEM micrographs revealed characteristic salt crystals and protein aggregates formed by the dried OF, which were observed for fresh samples and samples after six months post-deposition. Overall, this study demonstrated the great potential of SEM and ATR-FTIR spectroscopy for detecting OF traces on porous and nonporous substrates for up to six months for forensic purposes.