Determination and Confirmation of Psychedelic and Psychoactive Compounds in Botanical Materials by UPLC-ESI-MS/MS: Single Laboratory Validation.

Background: A multi-analyte method was developed for determination and confirmation of 26 natural and synthetic psychedelic and psychoactive compounds from botanical sources using a simple extraction procedure and UPLC-ESI-MS/MS analysis.

Objective: The objective of this work was to develop and validate the method for analysis of psychedelic and psychoactive compounds in botanicals (fungi, cactus, leaf), extract (brewed tea), synthetic and simulated finished product (using maltodextrin).

Methods: The method uses an acidified methanol solution for extraction and diphenhydramine as internal standard. Extracts are subjected to UPLC with tandem mass spectrometry using a phenyl-hexyl column and acidified mobile phase with acetonitrile gradient. Validation studies followed US Pharmacopeia <1225> and Official Methods of Analysis  SM Appendix K guidelines to assess specificity, accuracy (recovery), repeatability and intermediate precision, limits of detection and quantification, calibration curve linearity, system suitability, and robustness.

Results: Average recovery of spiked replicates in matrix and surrogate matrixes varied between 88-124% for analytes in fungi, cactus, and leaf matrixes; 81-106% in brewed tea matrix; and 81-174% across all 26 analytes in maltodextrin. Norbaeocystin and harmaline were the only analytes outside the acceptance criterion (79-126%) in maltodextrin. The 90% upper confidence limit on repeatability was ≤33% for all analytes and matrixes except for norbaeocystin. Repeatability precision for analysis of matrixes with native analytes met the acceptance criterion of ≤ 33% in all cases and the 90% upper confidence limit on intermediate precision was ≤35% in all cases. LOQ values fell below the lowest calibration standard (0.016 µg/mL) for all 26 analytes.

Conclusion: The method met the pre-determined acceptance criteria for recovery and precision in nearly all cases. Calibration curve linearity and system suitability measures were established.

Highlights: A method for 26 psychedelic/psychoactive compounds was validated for analysis of botanical (fungi, cactus, leaf), extract (brewed tea), synthetic and simulated finished product (using maltodextrin) using native and surrogate botanicals.