LCMS Detection and Characterization of In Vitro Metabolites of Isotonitazene, a Targeted Strategy for Novel Psychoactive Substance Control in Camel Racing.

Rationale: The use of benzimidazole-class novel psychoactive substances has significantly increased worldwide raising concerns about potential misuse and doping in animal sports such as camel racing. Understanding the metabolism of these substances is critical for developing reliable detection methods to ensure fair competition and animal welfare.

Methods: In vitro studies were conducted using homogenized camel liver samples to replicate metabolic processes. Metabolites were extracted and analyzed with a Thermo Fisher Orbitrap Exploris LCMS system. The acquired data were processed using the Compound Discoverer software for accurate detection and characterization of the metabolites.

Results: Through in vitro investigation, we have successfully identified nine metabolites in camels, which provide valuable insights into their metabolic pathways. The primary pathways for the formation of phase I metabolites were dealkylation and hydroxylation; the characterized metabolites can be used for long-term detection of the presence of isotonitazene.

Conclusion: The study employed advanced high-resolution accurate mass LC-MS techniques to detect and identify the in vitro metabolites of isotonitazene in camels. Isotonitazene is a highly active psychoactive substance with significant potential for abuse, particularly in racing contexts. These findings are crucial for developing effective doping control strategies, which will improve regulation and monitoring in camel racing events and promote animal welfare.