A Comparative In Vitro Metabolic Study of Butonitazene Using Camel Liver and Fungus Cunninghamella elegans for Doping...

Area of Science:

• Pharmacology
• Analytical Chemistry
• Veterinary Toxicology

Background:

• The emergence of potent synthetic opioids like butonitazene presents challenges for doping control.
• Butonitazene's structural uniqueness and high potency necessitate specific monitoring strategies, especially in animal racing.
• Understanding its metabolism is key to developing effective detection methods.

Purpose of the Study:

• To investigate the in vitro metabolism of butonitazene using camel liver and Cunninghamella elegans.
• To identify and characterize the metabolites of butonitazene.
• To assess the potential of these metabolites as biomarkers for doping control.

Main Methods:

• In vitro incubation of butonitazene with homogenized camel liver and Cunninghamella elegans.
• High-resolution liquid chromatography-mass spectrometry (LC-MS) analysis using a Thermo Fisher Orbitrap Exploris system.
• Identification and structural elucidation of butonitazene metabolites.

Main Results:

• 17 Phase I (N-dealkylation, O-dealkylation, hydroxylation) and 1 Phase II (sulfonation) metabolites were identified.
• Metabolite profiles were characterized using advanced LC-MS techniques.
• The identified metabolites show promise as long-term biomarkers for butonitazene detection.

Conclusions:

• Advanced LC-MS successfully identified in vitro metabolites of butonitazene in camels.
• The identified metabolites are crucial for developing robust doping control measures against butonitazene misuse.
• This research supports regulatory frameworks for animal welfare and sports integrity.