Rapid detection and validated quantification of psychoactive compounds in complex plant matrices by direct analysis in real time-high resolution mass spectrometry – Application to “Kava” psychoactive pepper products

Rationale: Classified by the UNODC as a top 20 plant of concern, Piper methysticum (also known as Kava) is being increasingly abused recreationally for its mind-altering effects. It is of significant forensic relevance to establish methods to rapidly identifyand quantify psychoactive compounds, especially those yet to be scheduled ascontrolled substances and which have exhibited various noteworthy health concerns. Methods: Direct analysis in real time high-resolution mass spectrometry (DART-HRMS) demonstrated the ability to detect a range of kavalactones in Pipermethysticum derived products and plant material with no sample preparation. Inaddition, a validated method using calibration curves developed with a deuteratedinternal standard was used for the quantification of the psychoactive moleculeyangonin in various products. Results: DART-HRMS detected the protonated masses of six major kavalactonesand three flavokavains in 18 commercial Kava products. A method consistent withFDA validation guidelines was established for the quantification of yangonin in thevarious complex matrices. Implementation of this method, with an LLOQ of 5 mg/mL, enabled successful quantification of yangonin in 16 Kava products.Concentrations for solid products ranged from 2.71 to 8.99 mg/g, while that forliquid products ranged from 1.03 to 4.59 mg/mL. Conclusions: Rapid identification and quantification of psychoactive smallmolecules in plant material can be accomplished using a validated DART-HRMSprotocol. This work illustrates an approach to qualitative and quantitative analysesof a wide variety of complex matrices derived from plants, and demonstrates thatthe commercially available products analyzed are P. methysticum derived and docontain psychoactive yangonin at quantifiable levels.