Detection of clobazam and its metabolites in urine during poisoning using HPLC-QqQ-MS/MS

Abstract

BACKGROUND: Determining the incidence of acute and lethal poisoning with clobazam remains an urgent task of analytical toxicology. The medicinal substance clobazam belongs to the group of benzodiazepines, which is included in the list of psychoactive substances whose turnover is limited because of its high toxicity profile in overdose and abuse. AIM: To propose a simple, reliable, and sensitive technique for the identification of clobazam and its metabolite in the urine by the modern high-performance liquid chromatography-tandem mass spectrometry (HPLC-QqQ-MS/MS). MATERIALS AND METHODS: We have described a simple and sensitive HPLC-QqQ-MS/MS method for the qualitative determination of clobazam in the urine. RESULTS: The retention time in the selected chromatography conditions for clobazam was 5.17 min, and its metabolite (norclobazam) in urine was 4.56 min. The base peaks for clobazam and norclobazam were 259 m/z and 245 m/z, respectively. CONCLUSION: For the first time, this study provides a validated method of chemical-toxicological examination for poisoning with clobazam by HPLC-MS/MS, tested both on a model mixture and real biological matrix of the patients urine after taking clobazam. This technique can be a confirmatory research method in addition to the clinical picture in forensic medical examination.