Novel qNMR Method to Quantify Psilocybin and Psilocin in Psychedelic Mushrooms

Psychedelic mushrooms of the Psilocybe genus contain the psychoactive tryptamines psilocybin and psilocin, compounds currently under clinical investigation for the treatment of depression and other psychiatric conditions. However, the accurate quantification of these alkaloids in fungal matrices remains analytically challenging. Here, we report an optimized extraction protocol and a robust, nondestructive quantification method based on quantitative nuclear magnetic resonance (qNMR) spectroscopy. Using a combination of ¹H- and ³¹P NMR, we achieved simultaneous detection and quantification of psilocin and psilocybin in dried Psilocybe cubensis samples with high accuracy and reproducibility. Our method revealed significant variability in tryptamine content and psilocybin-to-psilocin ratios among user-provided and laboratory-grown samples, underscoring the potential influence of storage conditions on alkaloid stability. Compared with conventional chromatographic approaches, qNMR offers a rapid and calibration-free alternative for the routine analysis of psychedelic fungi. This approach may facilitate quality control in emerging clinical and regulatory contexts of psychedelic mushrooms.