Differentiating Donor Age Groups Based on Raman Spectroscopy of Bloodstains for Forensic Purposes

[Image: see text] Developments in analytical chemistry technologies and portable instrumentation over the past decade have contributed significantly to a variety of applications ranging from point of care testing to industrial process control. In particular, Raman spectroscopy has advanced for analyzing various types of evidence for forensic purposes. Extracting phenotypic information (e.g., sex, race, age, etc.) from body fluid traces is highly desirable for criminal investigations. Identifying the chronological age (CA) of a blood donor can provide significant assistance to detectives. In this proof-of-concept study, Raman spectroscopy and chemometrics have been used to analyze blood from human donors, and differentiate between them based on their CA [i.e., newborns (CA of <1 year), adolescents (CA of 11–13 years), and adults (CA of 43–68 years)]. A support vector machines discriminant analysis (SVMDA) model was constructed, which demonstrated high accuracy in correctly predicting blood donors’ age groups where the lowest cross-validated sensitivity and specificity values were 0.96 and 0.97, respectively. Overall, this preliminary study demonstrates the high selectivity of Raman spectroscopy for differentiating between blood donors based on their CA. The demonstrated capability completes our suite of phenotype profiling methodologies including the determination of sex and race. CA determination has particular importance since this characteristic cannot be determined through DNA profiling unlike sex and race. When completed, the developed methodology should allow for phenotype profiling based on dry traces of body fluids immediately at the scene of a crime. The availability of this information within the first few hours since the crime discovery could be invaluable for the investigation.