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GC-MS and LC-(high-resolution)-MSn studies on the metabolic fate and detectability of camfetamine in rat urine

Abstract

Camfetamine (N-methyl-3-phenyl-norbornan-2-amine; CFA) belongs as amphetamine-type stimulant to the so-called new psychoactive substances. CFA is an analogue of fencamfamine, an appetite suppressant developed in the 1960s. The described effects of CFA are slight stimulation and increased vigilance and the side effects are tachycardia, paranoia, and sleeplessness. The aims of the presented work were to study the metabolic fate and the detectability of CFA in urine and to elucidate which cytochrome-P450 (CYP) isoenzymes are involved in the main metabolic steps. For metabolism studies, rat urine samples were isolated by solid-phase extraction without and after enzymatic cleavage of conjugates. The phase I metabolites were separated and identified after/without acetylation by gas chromatography-mass spectrometry (GC-MS) and/or liquid chromatography-high resolution-linear ion trap mass spectrometry (LC-HR-MSn), respectively, and the phase II metabolites by LC-HR-MSn. From the identified metabolites, the following main metabolic pathways were deduced: N-demethylation, aromatic mono or bis-hydroxylation followed by methylation of one hydroxy group, hydroxylation of the norbornane ring, combination of these steps, and glucuronidation and/or sulfation of the hydroxy metabolites. The N-demethylation was catalyzed by CYP2B6, CYP2C19, CYP2D6, and CYP3A4, the aromatic hydroxylation by CYP2C19 and CYP2D6, and the aliphatic hydroxylation was catalyzed by CYP1A2, CYP2B6, CYP2C19, and CYP3A4. Finally, the intake of a common user’s dose of CFA could be confirmed in rat urine using the authors’ GC-MS and the LC-MSn standard urine screening approaches via CFA and several metabolites, with the hydroxy-aryl CFA and the corresponding glucuronide being the most abundant.

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Acknowledgments

The authors thank Julia Dinger, Golo M. Meyer, Markus R. Meyer, Julian A. Michely, Carsten Schröder, Gabriele Ulrich, Armin A. Weber, and Carina S. D. Wink for support and/or helpful discussion.

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Correspondence to Hans H. Maurer.

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Welter, J., Kavanagh, P. & Maurer, H.H. GC-MS and LC-(high-resolution)-MSn studies on the metabolic fate and detectability of camfetamine in rat urine. Anal Bioanal Chem 406, 3815–3829 (2014). https://doi.org/10.1007/s00216-014-7796-6

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  • DOI: https://doi.org/10.1007/s00216-014-7796-6

Keywords

  • Designer drugs
  • Camfetamine
  • Metabolism
  • Cytochrome-P450
  • GC-MS
  • LC-(HR)-MSn