In vitro & In vivo Metabolism Studies of New Psychoactive Substances and Histopathological Patterns of Drug-Related Deaths: Comparative Forensic Analysis

This study investigates the metabolic pathways of four synthetic cathinones (3-CMC, 4-CMC, 4-BMC, NEPD) and N,N-Dimethyltryptamine (DMT) derivatives (DMT-Boc, DMT-isopropylcarbamate, DMT-pivaloylamide, DMT-THP), focusing on identifying biomarkers for forensic detection. Using human hepatocytes, liver microsomes, zebrafish embryos and, authentic human samples key metabolites were identified through liquid chromatography-high-resolution mass spectrometry. For synthetic cathinones, major metabolic reactions included N-deethylation, ketoreduction, and glucuronidation, with novel metabolic pathways observed in 3-CMC, 4-CMC and, NEPD. For DMT derivatives, N-demethylation, N-oxidation and, hydroxylation were the predominant metabolic reactions. Specific metabolites were proposed as markers of consumption for both synthetic cathinones and DMT derivatives. Histopathological analysis of autopsy cases showed significant tissue alterations in the heart, liver, lungs, and kidneys due to drug intake, including edema, fibrosis, and ischemic damage. These findings highlight the importance of combining toxicological and histopathological approaches in the study of NPSs and provide essential insights for forensic toxicologists, aiding in the detection of these substances in suspected abuse cases and highlighting the importance of comprehensive metabolite profiling in toxicological investigations.