Determination of time since deposition of bloodstains through NIR and UV–Vis spectroscopy – A critical comparison

Time elapsed since bloodstain deposition is a crucial aspect in forensic investigations, where non-destructive spectroscopic methods play a pivotal role. While extensive research has been conducted by UV–Vis spectroscopy, showcasing its utility in specific cases, there is still a paucity of studies based on NIR spectroscopy, which has the potential to overcome the limitations of the UV–Vis-based methods. To compensate for this disequilibrium, the present study aimed to evaluate the NIR applicability for estimating the age of forensic bloodstains and develop a performance comparison with UV–Vis spectroscopy methods. Capillary blood was sampled and subjected to a 16-day aging, during which it was repeatedly analyzed using both spectroscopic methods. Subsequently, chemometric analysis was applied to process the spectral data and independently assess the methods’ performance. Classical preprocessing transforms (i.e., Savitzky-Golay derivatives and SNV transform) were used together with more targeted strategies, such as class centering, whose benefit was highlighted by PCA. Lastly, PLS regression models were computed to evaluate the effectiveness of both spectroscopic methods in estimating the time elapsed since blood trace deposition. Comparable root mean square errors in prediction (RMSEP) – 40 and 55 h for UV–Vis and NIR spectroscopy, respectively – were observed for both techniques, featuring an improvement with respect to the existing literature for NIR spectroscopy. Data fusion strategies for a multi-instrumental platform were also explored, evaluating advantages and disadvantages of low-level and mid-level approaches. The results indicated that NIR spectroscopy integrated with adequate chemometric strategies deserves increased appreciation in forensic bloodstain dating.