Abstract
Mesembrine and mesembrenone are the main alkaloids of Sceletium tortuosum, a plant species that was used for sedation and analgesia by the KhoiSan, previously known as Hottentots, a tribe in South Africa. After fermentation, the obtained preparation called “Kanna” or “Kougoed” was used by chewing, smoking, or sniffing. Today, Kanna gains popularity by drug users as legal high. For monitoring such consumption, metabolism studies are mandatory because the metabolites are mostly the analytical targets, especially in urine. Therefore, the metabolism of both alkaloids was investigated in rat urine and pooled human liver preparations after several sample work-up procedures. As both alkaloids were not commercially available, they were isolated from plant material by Soxhlet extraction, and their identity confirmed by NMR. The metabolites were identified using gas chromatography–mass spectrometry (GC-MS) and liquid chromatography coupled to linear ion trap high resolution mass spectrometry (LC-HR-MSn). Both alkaloids were O- and N-demethylated, dihydrated, and/or hydroxylated at different positions. The phenolic metabolites were partly excreted as glucuronides and/or sulfates. Most of the phase I metabolites identified in rat urine could be detected also in the human liver preparations. After a common user’s low dose application of mesembrine, mainly the O- and N demethyl-dihydro, hydroxy, and bis-demethyl-dihydro metabolites, and in case of mesembrenone only the N-demethyl and the N-demethyl-dihydro metabolite could be detected in rat urine using the authors’ standard urine screening approaches (SUSA) by GC-MS or LC-MSn. Thus, it should be possible to monitor a consumption of mesembrine and/or mesembrenone assuming similar pharmacokinetics in humans.
Isolation and NMR identification of the Kanna alkaloids mesembrine and mesembrenone, followed by application to rats and incuabtion with human liver preparations for identification of their metabolites by GCMS and LC-(HR)-MSn
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Acknowledgments
The authors thank Jochen Beyer, Roland Hausmann, Tina Nikolaus, Markus R. Meyer, Julian Michely, Gabriele Ulrich, Carsten Schröder, and Armin A. Weber for their help and/or critical discussion.
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Published in the topical collection celebrating ABCs 13th Anniversary.
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Meyer, G.M.J., Wink, C.S.D., Zapp, J. et al. GC-MS, LC-MSn, LC-high resolution-MSn, and NMR studies on the metabolism and toxicological detection of mesembrine and mesembrenone, the main alkaloids of the legal high “Kanna” isolated from Sceletium tortuosum . Anal Bioanal Chem 407, 761–778 (2015). https://doi.org/10.1007/s00216-014-8109-9
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DOI: https://doi.org/10.1007/s00216-014-8109-9