Analytical and Bioanalytical Chemistry

, Volume 408, Issue 18, pp 4845–4856

First metabolic profile of PV8, a novel synthetic cathinone, in human hepatocytes and urine by high-resolution mass spectrometry

  • Madeleine J. Swortwood
  • Kayla N. Ellefsen
  • Ariane Wohlfarth
  • Xingxing Diao
  • Marta Concheiro-Guisan
  • Robert Kronstrand
  • Marilyn A. Huestis
Paper in Forefront

DOI: 10.1007/s00216-016-9599-4

Cite this article as:
Swortwood, M.J., Ellefsen, K.N., Wohlfarth, A. et al. Anal Bioanal Chem (2016) 408: 4845. doi:10.1007/s00216-016-9599-4

Abstract

Novel psychoactive substances (NPS) are ever changing on the drug market, making it difficult for toxicology laboratory methods to stay current with so many new drugs. Recently, PV8, a synthetic pyrrolidinophenone, was detected in seized products in Japan (2013), The Netherlands (2014), and Germany (2014). There are no controlled PV8 administration studies, and no pharmacodynamic and pharmacokinetic data. The objective was to determine PV8’s metabolic stability in human liver microsome (HLM) incubation and its metabolism following human hepatocyte incubation and high-resolution mass spectrometry (HRMS) with a Thermo Scientific Q-Exactive. Data were acquired with a full-scan data-dependent mass spectrometry method. Scans were thoroughly data mined with different data processing algorithms and analyzed in WebMetaBase. PV8 exhibited a relatively short 28.8 min half-life, with an intrinsic 24.2 μL/min/mg microsomal clearance. This compound is predicted to be an intermediate clearance drug with an estimated human 22.7 mL/min/kg hepatic clearance. Metabolic pathways identified in vitro included: hydroxylation, ketone reduction, carboxylation, N-dealkylation, iminium formation, dehydrogenation, N-oxidation, and carbonylation. The top three in vitro metabolic pathways were di-hydroxylation > ketone reduction > γ-lactam formation. Authentic urine specimen analyses revealed the top three metabolic pathways were aliphatic hydroxylation > ketone reduction + aliphatic hydroxylation > aliphatic carboxylation, although the most prominent peak was parent PV8. These data provide useful urinary metabolite targets (aliphatic hydroxylation, aliphatic hydroxylation + ketone reduction, aliphatic carboxylation, and di-hydroxylation) for forensic and clinical testing, and focus reference standard companies’ synthetic efforts to provide commercially available standards needed for PV8 biological specimen testing.

Graphical Abstract

Top four PV8 metabolites identified in vitro. Biotransformations highlighted in blue. Markush structures presented when exact location of biotransformation is unknown

Keywords

PV8 Novel psychoactive substances Metabolic profiling HRMS Hepatocytes Synthetic cathinones 

Supplementary material

216_2016_9599_MOESM1_ESM.pdf (823 kb)
ESM 1(PDF 822 kb)

Copyright information

© Springer-Verlag Berlin Heidelberg (outside the USA) 2016

Authors and Affiliations

  • Madeleine J. Swortwood
    • 1
  • Kayla N. Ellefsen
    • 1
    • 2
  • Ariane Wohlfarth
    • 1
    • 3
    • 4
  • Xingxing Diao
    • 1
  • Marta Concheiro-Guisan
    • 1
    • 5
  • Robert Kronstrand
    • 3
    • 4
  • Marilyn A. Huestis
    • 1
    • 6
  1. 1.Chemistry and Drug Metabolism, Intramural Research Program, National Institute on Drug AbuseNational Institutes of HealthBaltimoreUSA
  2. 2.Program in ToxicologyUniversity of Maryland BaltimoreBaltimoreUSA
  3. 3.Department of Forensic Genetics and Forensic ToxicologyNational Board of Forensic MedicineLinköpingSweden
  4. 4.Department of Medical and Health Sciences, Faculty of Health SciencesLinköping UniversityLinköpingSweden
  5. 5.Department of Sciences, John Jay College of Criminal JusticeCity University of New YorkNew YorkUSA
  6. 6.University of Maryland Baltimore School of MedicineSeverna ParkUSA