Metabolomics is the comprehensive analysis of small-molecule metabolites in living systems and is increasingly being applied in forensic science and health diagnostics. This review broadly integrates the foundational principles of metabolomics, key analytical techniques, and translational applications across forensic toxicology, postmortem interval estimation, and disease biomarker discovery. Advanced methodologies, such as mass spectrometry, nuclear magnetic resonance spectroscopy, and single-cell metabolomics, have knowingly enhanced sensitivity and resolution, enabling accurate detection of drug-related biomarkers, metabolic perturbations, and trauma-induced molecular signatures. Moreover, integrating metabolomics with cellular and molecular biology offers novel insights into disease pathophysiology, particularly in cancer, neurodegeneration, and metabolic disorders. Hence, emphasis is placed on the role of metabolite-mediated signaling and epigenetic regulation in bridging diagnostic gaps. The review delves deeper into recent advances in DNA-based phenotypic prediction, trace-based evidence collection strategies, and artificial-intelligence-driven analytical models. The conceptual and regulatory fundamentals of forensic and clinical metabolomics are compared in this review, exposing potential trends for transdisciplinary innovation.
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