Ayahuasca, a psychedelic beverage from the Amazon, is made by boiling Banisteriopsis caapi vine with DMT-containing plants. Using NMR and LC-MS/MS, this study analyzed both soluble and insoluble components of ayahuasca samples. For the first time, fructose was detected as a major component, and the alkaloid harmine was found in suspended solids. The major alkaloids identified were DMT, harmine, tetrahydroharmine, harmaline, and harmol. A new quantitative NMR method was developed and validated to simultaneously quantify these alkaloids.
An optimized extraction protocol and a quantitative nuclear magnetic resonance (qNMR) spectroscopy method enable accurate, nondestructive quantification of the psychoactive tryptamines psilocybin and psilocin in dried Psilocybe cubensis mushrooms. Using both 1H and 31P NMR, the method detects and measures both alkaloids simultaneously with high reproducibility. Analysis of user-provided and laboratory-grown samples reveals substantial variability in total tryptamine content and in the psilocybin-to-psilocin ratio, suggesting that storage conditions affect alkaloid stability. Compared to conventional chromatography, qNMR provides a rapid, calibration-free alternative for routine analysis, potentially supporting quality control in clinical and regulatory settings for psychedelic mushrooms.