HPLC Analysis of a Psilocybin-Based API from Psilocybe Mushrooms

Researchers at the Federal University of Campina Grande (Brazil) developed an active pharmaceutical ingredient (API) derived from Psilocybe mushrooms for application in the management of mental health disorders. The primary objectives of their work included the extraction of bioactive compounds, quantitative analysis, and finally the evaluation of physicochemical and biological properties. Following the extraction procedures, the team’s experiments achieved yields of approximately 20%, which showed the successful isolation of the compounds targeted. The presence of alkaloids was confirmed through phytochemical screening as well as validated by high-performance liquid chromatography (HPLC) analysis. A paper based on this research was published in ACS Omega (1).

In 2019, approximately 970 million people worldwide were determined to be affected by some form of mental disorder, which is characterized by clinical conditions that affect cognition, emotional regulation, or behavior, significantly impairing individual functioning and quality of life. This is a figure that increased significantly following the COVID-19 pandemic, with anxiety and depression reported as among the most commonly reported conditions (2).

Several therapeutic approaches have been proposed to mitigate or overcome such conditions for the wide number of those suffering from these conditions, including the use of antidepressants, neuromodulation techniques, and psychotherapy. Clinical trials have reported improvements in mental health through psychedelic-assisted psychotherapy, involving such substances as MDMA (3,4-methylenedioxymethamphetamine) for post-traumatic stress disorder (PTSD), LSD (lysergic acid diethylamide) for anxiety, ibogaine for alcoholism, DMT (dimethyltryptamine) for depression, and psilocybin (a naturally occurring tryptamine alkaloid and investigational drug found in over 200 species of mushroom), with the latter showing broad-spectrum potential because of its favorable safety and efficacy profiles (3-6).

Recent studies concerning Psilocybe cubensis have shown that extracts obtained from this fungus enhanced memory and increased levels of brain-derived neurotrophic factor (BDNF) in the hippocampus of animal models (7). Proteomic analyses have found peptides with antimicrobial activity present; these peptides have the potential to inhibit the growth of Staphylococcus aureus (8). Furthermore, there have been recent investigations which evaluated the effects of psychedelic compounds present in P. cubensi on microglial cells, demonstrating modulation of the microglial immune response (9). Psychedelic compounds, including psilocybin, have also shown to be effective in the alleviation of a variety of symptoms associated with post-COVID-19 syndrome, including olfactory dysfunction, cognitive deficits, excessive sleepiness, sleep disturbances, and psychological symptoms (10).

The API the researchers obtained from the mushrooms analyzed exhibited thermal and spectroscopic characteristics consistent with those of edible mushroom extracts and allowed the team to gain a comprehensive understanding of its physicochemical behavior. Analytical assays demonstrated high solubility, low toxicity, and compliance with acceptable limits for heavy metal content and microbial load, including aerobic microorganisms, fungi, and yeasts, as well. Their quantitative HPLC analysis revealed psilocybin and psilocin contents of 3.26 and 0.34%, respectively, which supported the formulation of drug delivery systems with standardized concentrations of psilocybin. The results showed that the API extracted was stable, highly pure, and compatible with polymeric matrices for controlled release applications. In addition, the assays confirmed high solubility in polar solvents and minimal risk of adverse effects (1).

The researchers stated that their study showed that the development of a psilocybin- and psilocin-based API is not only possible but signifies an improvement in the management of mental disorders and an auspicious option in the pursuit of innovative therapeutic treatments (1).

Read more on this topic: Applications of Untargeted Chemical Analysis to Determine Psilocybin and Psilocin Content in Multiple Psilocybe cubensis Mushroom Strains Using LC–MS/MS: An Interview with Kevin Schug

References

1. Pereira Galdino, T.; Bisneto, A. B. M.; Pedro, M. D. S. et al. Multianalytical Investigation of Psilocybe cubensis Mushrooms: Physicochemical Characterization and Biological Evaluation of Psilocybin and Psilocin Compounds. ACS Omega 2025, 10 (38), 44173-44189. DOI: 10.1021/acsomega.5c05606
2. Mental disorders. World Health Organization website . 2022. https://www.who.int/news-room/fact-sheets/detail/mental-disorders (accessed by the authors of [1] 2025-04-01)
3. Siegel, A. N.; Meshkat, S.; Benitah, K. et al. Registered Clinical Studies Investigating Psychedelic Drugs for Psychiatric Disorders. J. Psychiatr. Res. 2021, 139, 71– 81, DOI: 10.1016/j.jpsychires.2021.05.019
4. dos Santos Gonçalves, W.; Lassen, R. D. H.; Appolinario, J. C. et al. Diagnosis and Therapeutic Strategy in Treatment-Resistant Depression. Med. Cien. e Arte 2022, 1 (3), 94– 104
5. Tavares, L. A.; Salvoni, J. F.; Zambrana, B. C. d. S. D. et al. Therapeutic Approaches in Suicide Prevention: Evidence-Based Strategies. Brazilian J. Implantol. Health Sci. 2023, 5 (5), 4719– 4732. DOI: 10.36557/2674-8169.2023v5n5p4719-4732
6. Psilocybin. Wikipedia.https://en.wikipedia.org/wiki/Psilocybin (accessed 2025-10-07)
7. Ghaffarzadegan, R.; Karimi, M.; Hedayatjoo, B. et al. The Effect of Psilocybe cubensis on Spatial Memory and BDNF Expression in Male Rats Exposed to Chronic Unpredictable Mild Stress. J. Psychoact. Drugs 2024, 1– 13, DOI: 10.1080/02791072.2024.2428241
8. Karthiyayini, B.; Karthick Raja, A.; Arthi, S. et al. Proteomic Analysis of Psychedelic Mushroom Isolate and Exploring Potential Antimicrobial Peptides Against Bacteria. Natural Product Research 2024, 1. DOI: 10.1080/14786419.2024.2423045
9. Laabi, S.; LeMmon, C.; Vogel, C. et al. Psilocybin and Psilocin Regulate Microglial Immunomodulation and Support Neuroplasticity via Serotonergic and AhR Signaling. International Immunopharmacology 2025, 159, 114940. DOI: 10.1016/j.intimp.2025.114940
10. Low, Z. X. B.; Yong, S. J.; Alrasheed, H. A. et al. Serotonergic Psychedelics as Potential Therapeutics for Post-COVID-19 Syndrome (or Long COVID): A Comprehensive Review. Progress in Neuro-Psychopharmacology and Biological Psychiatry 2025, 137, 111279. DOI: 10.1016/j.pnpbp.2025.111279