A highly sensitive method for measuring psilocin and psilocybin in hair samples from magic mushroom users was developed using ultra-high pressure liquid chromatography coupled with tandem mass spectrometry. The technique achieved a detection limit of 1 pg/mg and a quantification limit of 5 pg/mg, with linear calibration from 5 to 500 pg/mg. In two authentic hair samples, psilocin concentrations were 161 and 150 pg/mg, while psilocybin was not detected. The method also analyzed psilocin and psilocybin distribution in seven hallucinogenic mushrooms. This represents the first measurement of psilocin in hair from hallucinogenic mushroom consumers and the most sensitive quantitative method for these compounds in hair.
4-Acetoxy-N,N-dimethyltryptamine (4-AcO-DMT, psilacetin) is a synthetic psychedelic that may act as a precursor to psilocin, but its metabolism was poorly understood. Incubating 4-AcO-DMT with pooled human liver microsomes produced 15 metabolites: 12 from phase I and 3 from phase II reactions. Transformations included hydrolysis, hydroxylation, N-demethylation, oxidation, and glucuronic acid conjugation. The hydrolysis product was the most abundant. For forensic detection of 4-AcO-DMT use, the beta-hydroxylation metabolite (M2-1) is recommended as a biomarker. These findings may help predict in vivo metabolism and assist drug testing.