Skip to main content
SpringerLink
Log in

In vivo metabolism of the new synthetic cannabinoid APINAC in rats by GC–MS and LC–QTOF-MS

  • Original Article
  • Published:
Forensic Toxicology Aims and scope Submit manuscript

Abstract

The recent appearance of APINAC (AKB-57, ACBL(N)-018, adamantan-1-yl 1-pentyl-1H-indazole-3-carboxylate) in the market of the so-called novel psychoactive substances resulted in the need of defining its characteristics and searching its metabolites for subsequent detection in biological samples. The structure of the APINAC molecule has great similarity to the molecules of other synthetic cannabinoids. Here we report on the in vivo metabolism of APINAC using rats as an experimental model. Rat urine samples were analyzed by using gas chromatography–mass spectrometry and liquid chromatography–high resolution mass spectrometry. Data were acquired via time-of-flight mass scan, followed by Auto MS and triggered product ion scans. The predominant metabolic pathway for APINAC was ester hydrolysis yielding a wide variety of N-pentylindazole-3-carboxylic acid metabolites and 1-adamantanol metabolites. Ten metabolites for APINAC were identified, with the majority generated by hydroxylation, carbonylation, and carboxylation with or without glucuronidation. Therefore, in vivo metabolic profiles in rats were generated for APINAC. N-Pentylindazole-3-carboxylic acid, hydroxylated N-pentylindazole-3-carboxylic acid, and 1-adamantanol are likely the best targets to incorporate into analytical screening methods for drugs analysis. The presented mass spectra and retention time data may be useful for detection of these compounds in human urine.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Reggio PH (ed) (2009) The cannabinoid receptors. Humana Press, New York

    Google Scholar 

  2. EMCDDA (2016). Thematic paper—understanding the ‘Spice’ phenomenon. http://www.emcdda.europa.eu/attachements.cfm/att_80086_EN_Spice%20Thematic%20paper%20%E2%80%94%20final%20version.pdf. Accessed May 2016

  3. Uchiyama N, Kikura-Hanajiri R, Kawahara N, Goda Y (2009) Identification of a cannabimimetic indole as a designer drug in a herbal product. Forensic Toxicol 27:61–66

    Article  CAS  Google Scholar 

  4. EMCDDA (2016) Perspectives on drugs. Synthetic cannabinoids in Europe. http://www.emcdda.europa.eu/attachements.cfm/att_212361_EN_Synthetic%20cannabinoids_updated2016.pdf. Accessed Sept 2016

  5. Courts J, Maskill V, Gray A, Glue P (2016) Signs and symptoms associated with synthetic cannabinoid toxicity: systematic review. Australas Psychiatry 24:598–601

    Article  PubMed  Google Scholar 

  6. Lee JH, Park HN, Leem TS, Jeon JH, Cho S, Lee J, Baek SY (2017) Identification of new synthetic cannabinoid analogue APINAC (adamantan-1-yl 1-pentyl-1H-indazole-3-carboxylate) with other synthetic cannabinoid MDMB(N)-Bz-F in illegal products. Forensic Toxicol 35:45–55

    Article  CAS  Google Scholar 

  7. Diao X, Huestis MA (2017) Approaches, challenges and advances in metabolism of new synthetic cannabinoids and identification of optimal urinary marker metabolites. Clin Pharmacol Ther 101:239–253

    Article  CAS  PubMed  Google Scholar 

  8. Vikingsson S, Josefsson M, Gréen H (2015) Identification of AKB-48 and 5F-AKB-48 metabolites in authentic human urine samples using human liver microsomes and time of flight mass spectrometry. J Anal Toxicol 39:426–435

    Article  CAS  PubMed  Google Scholar 

  9. Sobolevsky T, Prasolov I, Rodchenkov G (2015) Study on the phase I metabolism of novel synthetic cannabinoids, APICA and its fluorinated analogue. Drug Test Anal 7:131–142

    Article  CAS  PubMed  Google Scholar 

  10. Grigoryev A, Kavanagh P, Melnik A (2012) The detection of the urinary metabolites of 3-[(adamantan-1-yl)carbonyl]-1-pentylindole (AB-001), a novel cannabimimetic, by gas chromatography–mass spectrometry. Drug Test Anal 4:519–524

    Article  CAS  PubMed  Google Scholar 

  11. Wohlfarth A, Gandhi AS, Pang S, Zhu M, Scheidweiler KB, Huestis MA (2014) Metabolism of synthetic cannabinoids PB-22 and its 5-fluoro analog, 5F-PB-22, by human hepatocyte incubation and high-resolution mass spectrometry. Anal Bioanal Chem 406:1763–1780

    Article  CAS  PubMed  Google Scholar 

  12. Diao X, Scheidweiler KB, Wohlfarth A, Pang S, Kronstrand R, Huestis MA (2016) In vitro and in vivo human metabolism of synthetic cannabinoids FDU-PB-22 and FUB-PB-22. AAPS J 18:455–464

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Diao X, Wohlfarth A, Pang S, Scheidweiler KB, Huestis MA (2016) High-resolution mass spectrometry for characterizing the metabolism of synthetic cannabinoid THJ-018 and its 5-fluoro analog THJ-2201 after incubation in human hepatocytes. Clin Chem 62:157–169

    Article  CAS  PubMed  Google Scholar 

  14. Gandhi AS, Zhu M, Pang S, Wohlfarth A, Scheidweiler KB, Liu H, Huestis MA (2013) First characterization of AKB-48 metabolism, a novel synthetic cannabinoid, using human hepatocytes and high-resolution mass spectrometry. AAPS J 15:1091–1098

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Grigoryev A, Savchuk S, Melnik A, Moskaleva N, Dzhurko J, Ershov M, Nosyrev A, Vedenin A, Izotov B, Zabirova I, Rozhanets V (2011) Chromatography-mass spectrometry studies on the metabolism of synthetic cannabinoids JWH-018 and JWH-073, psychoactive components of smocking mixtures. J Chromatogr B 879:1126–1136

    Article  CAS  Google Scholar 

  16. Grigoryev A, Melnik A, Savchuk S, Simonov A, Rozhanets V (2011) Gas and liquid chromatography–mass spectrometry studies on the metabolism of synthetic phenylacetylindole cannabimimetic JWH-250, psychoactive component of smoking mixtures. J Chromatogr B 879:2519–2526

    Article  CAS  Google Scholar 

  17. Diao X, Deng P, Xie C, Li X, Zhong D, Zhang Y, Chen X (2013) Metabolism and pharmacokinetics of 3-n-butylphthalide (NBP) in humans: the role of cytochrome P450s and alcohol dehydrogenase in biotransformation. Drug Metab Dispos 41:430–433

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Dr. Andrey Grigoriev for data acquisition in this study. In addition, Dr. Andrey Grigoriev is kindly acknowledged for his valuable comments on the study and the manuscript. This work was supported by grants from the University Research Fund, Sechenov First Moscow State Medical University.

Author information

Authors and Affiliations

  1. Institute of Pharmacy and Translational Medicine, Sechenov First Moscow State Medical University, 2-4 Bolshaya Pirogovskaya St., Moscow, 119991, Russia

    Sergey Savchuk, Svetlana Appolonova & Ksenia Shestakova

  2. Russian Center of Forensic Medical Expertise, Polikarpova St., Moscow, 125284, Russia

    Sergey Savchuk

  3. Internet-portal “Forum Forensic Experts in Russia”, Salekhard, Russia

    Alexander Pechnikov

  4. Laboratory for Forensic Chemistry at Clinical Psychoneurological Hospital, Nizhnevartovsk, Russia

    Liliay Rizvanova

  5. Unit of Forensic Medicine, Department of Diagnostics and Public Health, University of Verona, Policlinico “G.B. Rossi”, Piazzale L.A. Scuro 10, 37134, Verona, Italy

    Franco Tagliaro

Authors
  1. Sergey Savchuk
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Svetlana Appolonova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alexander Pechnikov
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Liliay Rizvanova
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ksenia Shestakova
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Franco Tagliaro
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Svetlana Appolonova.

Ethics declarations

Conflict of interest

The authors declare that there are no financial or other relations that could lead to a conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines were followed for the care and use of the animals. This study did not involve samples collected from human participants.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Savchuk, S., Appolonova, S., Pechnikov, A. et al. In vivo metabolism of the new synthetic cannabinoid APINAC in rats by GC–MS and LC–QTOF-MS. Forensic Toxicol 35, 359–368 (2017). https://doi.org/10.1007/s11419-017-0364-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11419-017-0364-y

Keywords

Search

Navigation