Automated and semi-automated screening, LCMSMS confirmation and human hepatocyte high-resolution metabolism studies of synthetic cannabinoids

Designer drug availability changes rapidly. Assays are needed to identify synthetic cannabinoids in human urine, but metabolites are frequently unknown. This research evaluates available screen performance, develops sensitive and specific LCMSMS confirmation assays, and utilizes human hepatocyte cultures and high-resolution (HR) mass spectrometry to identify unique urinary targets. Homogeneous (Immunalysis), and heterogeneous (Randox; National Medical Services (NMS)) screening assays were evaluated with authentic urine specimens. An LCMSMS qualitative confirmation method for synthetic cannabinoids (JWH-018, JWH-073, JWH-081, JWH-122, JWH-200, JWH-210, JWH-250, AM 2201 and RCS-4) and 20 metabolites in human urine was developed. Specimens were hydrolyzed and protein precipitated, followed by single MRM transition monitoring or survey scan that triggered an enhanced product ion scan at 3 different collision energies on the ABSciex 5500 Qtrap. 2500 specimens were screened by Randox, Immunalysis, NMS, and LCMSMS assays for 19 hydroxyalkyl, hydroxyindole and carboxy metabolites of 9 synthetic cannabinoids. Positive criteria included retention time, purity of fit, and presence of 3 ions, including the molecular ion. Confirmation based on full spectra increased identification confidence. 285 samples were LCMSMS positive for synthetic cannabinoids. Immunalysis and NMS assays had high sensitivities (87.7 and 84.9 per cent) and specificities (98.7 and 97.7 per cent) at a 5ng/mL cutoff; Randox identified almost all positive specimens (97.9 per cent sensitivity), but had 1149 presumptive positive unconfirmed specimens (48.1 per cent specificity). Possible reasons for low specificity, and advantages and disadvantages of approaches are discussed. Human hepatocyte cultures and HR mass spectrometry identified new synthetic cannabinoids metabolites to improve emerging drug identification. Identification of new synthetic cannabinoids is a difficult and critical laboratory problem.

Language

  • English

Media Info

  • Pagination: 356-61
  • Monograph Title: 20th International Conference on Alcohol, Drugs and Traffic Safety Conference Proceedings, 25-28 August 2013, Brisbane Convention and Exhibition Centre, Brisbane, Australia

Subject/Index Terms

Filing Info

  • Accession Number: 01495684
  • Record Type: Publication
  • Source Agency: ARRB
  • Files: ITRD, ATRI
  • Created Date: Oct 17 2013 10:12AM