Psilocybe mushrooms: extraction, identification, and quantification of psychedelic tryptamines

Abstract

Introduction: “Magic mushrooms”, belonging to the genus Psilocybe, contain the tryptamines psilocybin and psilocin, compounds with recognised therapeutic potential for mental health (Blei et al., 2020). Psilocybin is generally more abundant, with concentrations in dried mushrooms ranging from 0.5% to 1.5% (Goff et al., 2024). Their use for religious, recreational, and therapeutic purposes, along with their growing popularity and potential harm, has led to the development of specific regulatory frameworks (Tylš et al., 2014), highlighting their increasing importance in forensic sciences.

Objective: Development of fast methods for the extraction, identification, and quantification of psilocin and psilocybin present in Psilocybe mushrooms.

Methods: An Agilent 1260 Infinity II HPLC-DAD system with a Poroshell 120 EC-C18 3.0 × 150 mm, 2.7 µm column was used for analytical determinations, with ultraviolet detection at 220 nm. The final extraction protocol involved pulverisation of mushrooms in a mortar and pestle on ice, followed by two 15-minute extractions using methanol containing 10% water (5 mL + 5 mL for 50.0 mg of mushroom) through dynamic maceration by stirring.

Results: To optimise the extraction method, several parameters were considered, including extraction time, number of extractions, agitation rate, solvent used, and the solvent-to-dry material ratio. The concentrations of psilocybin and psilocin were 1.976% and 0.101%, respectively, expressed as % (w/w) in the mushroom. A Thin Layer Chromatography method using Ehrlich's reagent as the revealing solution was established to enable rapid and well-resolved identification of psilocybin and psilocin.

Conclusions: This work has developed novel extraction and analytical methods for tryptamines suitable for forensic applications.