QUANTITATIVE NUCLEAR MAGNETIC RESONANCE FOR THE QUANTIFICATION OF PSYCHEDELICS IN AYAHUASCA AND MUSHROOM SAMPLES

The resurgence of psychedelic research has revealed potential clinical applications for psilocybin and other tryptamines in the context of assisted psychotherapy for the treatment of neuropsychiatric disorders such as major depressive disorder (1), (2). In particular, psychedelic mushrooms of the Psilocybe genus, historically used for medicinal, spiritual, and religious purposes (3), have gained renewed attention due to the presence of psilocybin (PSB) and its active metabolite, psilocin (PS).

Similarly, ayahuasca, a decoction made by boiling two plant species—Banisteriopsis caapi and Psychotria viridis has been traditionally used in indigenous settings for religious and medicinal purposes (4). This brew, known to induce altered states of consciousness, has sparked the interest of the scientific community due to its potential efficacy in controlled clinical settings for the treatment of disorders such as depression.

To date, clinical trials have predominantly involved the use of pure psilocybin, and it is unknown whether interventions with whole mushrooms can achieve comparable safety and efficacy. A similar question arises regarding the use of ayahuasca versus farmahuasca (pure DMT and beta-carbolines). To address these questions, it is essential to have robust and reproducible analytical methodologies capable of accurately quantifying psychoactive alkaloid content in different natural matrices. These methods are also essential for the advancement of natural product research, enabling the study of other plant and fungal species that may contain these compounds.

In this study, we developed an analytical methodology based on Quantitative Nuclear Magnetic Resonance (qNMR) spectroscopy for the determination of DMT and beta-carbolines in ayahuasca samples, as well as psilocybin and psilocin in psychedelic mushroom extracts. Unlike conventional chromatographic methods, which often rely on variable extraction protocols and lack standardization, NMR spectroscopy offers significant advantages: non-destructive analysis, simultaneous detection of multiple metabolites, and linear quantification over a wide dynamic range.

The development and optimization of the method are presented, highlighting its speed, reproducibility, and robustness. To demonstrate its applicability, we analyzed extracts of mushrooms collected in Uruguay belonging to the genera Psilocybe, Conocybe, and Panaeolus, as well as ayahuasca samples donated by users.

This study represents the first application of quantitive NMR spectroscopy to the analysis of psychedelic mushrooms and ayahuasca, demonstrating its potential for comprehensive determination of secondary metabolites. The ability to simultaneously detect and quantify multiple components within a single spectrum makes this method a valuable tool for advancing research into the pharmacological properties and phytochemical diversity of psychoactive natural products.

REFERENCES

(1) Davis, A. K.; Barrett, F. S.; May, D. G.; Cosimano, M. P.; Sepeda, N. D.; Johnson, M. W.; Finan, P. H.; Griffiths, R. R. JAMA Psychiatry 2020, 78 (5), 481.

(2) Palhano-Fontes F, Barreto D, Onias H, et al. Psychological Medicine 2018, 49 (4), 655–663.

(3) Ott, J. Pharmacotheon: Entheogenic Drugs, Their Plant Sources and History, 2_nd ed., 1996.

(4) Labate, B. C.; Cavnar, C. The therapeutic use of ayahuasca; 2013.

Acknowledgements:

Facultad de Química, Universidad de la República (Uruguay), the National Research and Innovation Agency (ANII), and the Interdisciplinary Center for Psychedelic Studies (ARCHE)