Forensic applications of postmortem protein analysis in estimating the time since death: a systematic review

Estimating the postmortem interval (PMI) represents one of the most complex challenges in forensic medicine, especially beyond the first hours after death, when traditional methods become unreliable. In recent years, attention has progressively shifted toward the study of biomolecular alterations, with particular interest in protein alterations. This systematic review has been conducted according to PRISMA guidelines, leading to the enrollment of 46 articles published between 2014 and 2025, regarding postmortem protein modifications in human and animal tissues and according to specific inclusion criteria. Skeletal muscle was the most studied tissue, due to its abundance and ease of sampling, with markers such as tropomyosin, desmin, vinculin, α-actin, and GAPDH showing well-defined degradation patterns that are little influenced by external factors such as the cause of death or environmental conditions. Differences related to muscle type, the presence of lesions, and intra and/or interindividual variables were found. Furthermore, proteomic analysis of other tissues, particularly heart and bone, highlighted distinct behaviors and potential applications for longer time periods. Therefore, trying to combine the data acquired from multiple proteins and tissues, could be a promising approach for increasing the reliability and accuracy for PMI estimation.