Optimization of Psilocybin Extraction from Psilocybe cubensis Mushrooms and Characterization of the Fungal Extract

Psilocybin is a molecule with significant potential for the treatment of mental disorders such as depression, anxiety, post-traumatic stress disorder, and substance abuse. In this study, it was explored the extraction of psilocybin from Psilocybe cubensis mushrooms using a factorial design approach, along with the physicochemical and phytochemical characterization of the extracted material, aiming to optimize the process. The results indicate that the optimal extraction conditions were achieved using an acidified ethanol solvent (without water addition) at pH 2 and at temperature of 25 °C. The maximum yield obtained was 50.03 mg of psilocybin g-1 of extract (ca. 1% psilocybin), which exceeds the values reported in the literature for similar studies, demonstrating the efficiency of the proposed method. Through liquid chromatography-mass spectrometry (LC-MS/MS) analysis, six compounds were annotated: the indole alkaloids psilocybin and psilocin, previously reported in the P. cubensis genus, and four additional compounds, sn-glycero-3-phosphocholine, norvaline, tetronomycin and N-(tetradecanoyl)-sphinganine, which have not previously been reported in the Psilocybe genus.