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Metabolite elucidation of 2-fluoro-deschloroketamine (2F-DCK) using molecular networking across three complementary in vitro and in vivo models.

Thomas Gicquel, Romain Pelletier, Camille Richeval, Alexandr Gish, Florian Hakim, Pierre-Jean Ferron, Vadim Mesli, Delphine Allorge, Isabelle Morel, Jean-Michel Gaulier

Drug testing and analysis January 1, 2022 DOI: 10.1002/dta.3162 via PubMed

Summary

AI-generated from the abstract

Thirteen metabolites of the dissociative drug 2-fluoro-deschloroketamine (2F-DCK) were produced in vitro using human liver microsomes and HepaRG liver cells. Seven additional metabolites, including three Phase II conjugates, were identified in post-mortem bile and urine from a fatal case. Molecular networking helped compare the two in vitro models, which proved complementary. The authors propose that nor-2F-DCK (mass-to-charge 208.1137) and a hydrogenated metabolite (224.1443) are reliable markers for detecting 2F-DCK use in high-resolution mass spectrometry libraries.

Study at a glance

Characteristics In vitro metabolism study with post-mortem case application Peer reviewed
Population Human liver microsomes, HepaRG cell line, and post-mortem bile and urine from one fatality
Intervention 2F-DCK
Dose 100 and 500 μM for HLMs; 20 μM for HepaRG cells
Duration Up to 1 h for HLMs; 8 and 24 h for HepaRG cells
Keywords 2f-dck Heparg cell line Metabolism Molecular networking New psychoactive substances
Citations 32
Key finding Thirteen in vitro and seven in vivo metabolites of 2F-DCK were identified, with nor-2F-DCK and a hydrogenated metabolite proposed as reliable markers for detection.

Abstract

This work first aims to investigate metabolites of 2-fluoro-deschloroketamine (2F-DCK), a new arylcyclohexylamine derivatives (a group of dissociative ketamine-based substances) using two in vitro experimental approaches, and to compare obtained results by means of molecular networking. Metabolites of 2F-DCK were investigated using both human liver microsomes (HLMs) and hepatic (HepaRG) cell line incubates using molecular networking approach: 2F-DCK pure substance was incubated with HLMs for up to 1 h at two concentrations (100 and 500 μM) and with HepaRG cells for two time periods (8 and 24 h) at one concentration (20 μM). In vitro obtained results were subsequently applied to a 2F-DCK-related fatality case. In vitro-produced metabolites were investigated using high-resolution accurate mass spectrometry using Orbitrap mass analyzer technology. Thirteen metabolites were in vitro produced and several metabolic pathways can be postulated. Seven additional metabolites were found in post-mortem samples (bile and urine) of the case, comprising three Phase II metabolites, which appear to be minor in vivo metabolites. HLMs and HepaRG cell models appear to be complementary and obtained data allowed the identification of several specific 2F-DCK metabolites in biological samples. In practical terms, observed metabolic ratios suggested that nor-2F-DCK (208.1137 m/z) and a hydrogenated metabolite (224.1443 m/z) could be proposed as reliable metabolites to be recorded in HRMS libraries in order to improve detection of 2F-DCK use.

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