Caracterização fitoquímica, otimização da extração e quantificação dos princípios ativos do cogumelo Psilocybe cubensis

Abstract

Mushrooms of the genus Psilocybe, whose popular name is “magic mushroom”, have hallucinogenic effects due to the presence of indolic alkaloids in their composition, and have been studied for their application in the treatment of mental disorders such as depression and anxiety. The aims of this study were therefore to carry out quality control of the Psilocybe cubensis mushroom and its extract, extract the indolic alkaloids in high yields and obtain psilocin by dephosphorylating psilocybin. The dried fungal material was crushed, sieved and extracted with ethanol. The mushroom powder was measured for pH in water, which was 6.68, for granulometry, as it is a semi-fine powder with a density of 5.5 g/mL, and for cadmium and lead content, both below 0.1 μg/g, and the ash content was 7.4%. Phytochemical tests were carried out with positive results for various classes of metabolites in the extract. A chromatographic method was developed and validated for the quantification of alkaloids in accordance with RDC standard N°166, using a gradient of acetonitrile:acidified water 5:95 as the mobile phase. The detection limit was 1.58 mg/L for psilocybin and 1.70 mg/L for psilocin, and the quantification limit was 4.80 mg/L for psilocybin and 5.17 mg/L for psilocin. The matrix effect and the robustness of the method were also determined and were not influential. In order to optimize the extraction of psilocybin, a factorial design 23 was carried out and then a BoxBehnken design 24 with the variables of time, pH, quantity of extractions and volume of extracting solvent, the best condition for extraction being when the solvent consisted solely of ethanol with 2% acetic acid, and five consecutive extractions with 4 mL for 100 mg of powder, in which contents greater than 3.5% of psilocybin per dry mass of extract were obtained. The extract was tested against strains of Staphylococcus aureus ATCC 13656 (Gram positive), Escherichia coli ATCC 25422 (Gram negative) and Candida Albicans ATCC 76645 (Fungus), using the micro dilution method in a 96-well plate with a range of 25 - 25000 mg/L, but proved to be inactive against these strains. Finally, the dephosphorylation of psilocybin was achieved using acidified water in the presence of chloroform at a temperature of 120°C, with a yield of 16%.