-Methyltryptamine (-MT) Metabolite Profiling in Human Hepatocyte Incubations and Postmortem Urine and Blood

alpha-MT is a hallucinogenic and stimulant tryptamine that was involved in several overdose fatalities in the United States and Europe. Analytical toxicology, and particularly the identification of metabolite biomarkers in biological samples, often is the only way to prove tryptamine use in clinical and forensic caseworks. We aimed to identify optimal alpha-MT metabolite biomarkers of consumption in humans. We identified alpha-MT metabolites in 10-donor-pooled human hepatocyte incubations and postmortem urine and blood from an alpha-MT overdose case using in silico metabolite predictions, liquid chromatography high-resolution-tandem mass spectrometry (LC-HRMS/MS), and software-assisted data mining. Nine metabolites were identified in vitro and eight additional metabolites were found in urine; five metabolites were found in blood. Metabolic transformations were hydroxylation, O-sulfation, O-glucuronidation, N-glucuronidation, and N-acetylation, consistent with the metabolism of structural analogues. The findings in hepatocyte incubations and postmortem samples were consistent, proving the in vitro model suitability. We suggest alpha-MT, hydroxy-alpha-MT glucuronide, and two hydroxy-alpha-MT sulfates as biomarkers of alpha-MT use in non-hydrolyzed urine; we suggest alpha-MT, two hydroxy-alpha-MT sulfates and N-acetyl-alpha-MT as biomarkers of alpha-MT use in blood. Further studies on alpha-MT clinical and forensic caseworks with different doses and routes of administration are necessary to better explore alpha-MT metabolism.