Postmortem interval (PMI) estimation is a challenge of utmost importance in forensic daily practice. Traditional methods face limitations in accuracy and reliability, particularly for advanced decomposition stages. Recent advances in "omics" sciences, providing a holistic view of postmortem biochemical changes, offer promising avenues for overcoming these challenges. This systematic review aims at investigating the role of mass-spectrometry-based "omics" approaches in PMI estimation to elucidate molecular mechanisms underlying predictable time-dependent biochemical alterations occurring after death. A systematic search was performed, adhering to PRISMA guidelines, through "free-text" protocols in the databases PubMed, SCOPUS and Web of Science. The inclusion criteria were as follows: experimental studies analyzing, as investigated samples, animal or human corpses in toto or in parts and estimating PMI through MS-based untargeted omics approaches, with full texts in the English language. Quality assessment was performed using STROBE and ARRIVE critical appraisal checklists. A total of 1152 papers were screened and 26 included. Seventeen papers adopted a proteomic approach (65.4%), nine focused on metabolomics (34.6%) and two on lipidomics (7.7%). Most papers (57.7%) focused on short PMIs (<
7 days), the remaining papers explored medium (7-120 days) (30.77%) and long PMIs (>
120 days) (15.4%). Muscle tissue was the most frequently analyzed substrate (34.6% of papers), followed by liver (19.2%), bones (15.4%), cardiac blood and leaking fluids (11.5%), lung, kidney and serum (7.7%), and spleen, vitreous humor and heart (3.8%). Predictable time-dependent degradation patterns of macromolecules in different biological substrates have been discussed, with special attention to molecular insights into postmortem biochemical changes.