A preliminary gas chromatography-mass spectrometry-based metabolomics study of rats ingested diazepam or clonazepam

Drug-facilitated sexual assault (DFSA) is a sexual act in which the victim is unable to give or rescind consent due to alcohol or drug intoxication, which involved the abuse of benzodiazepines around the world. Conventional techniques used for the analysis of benzodiazepines have the limitation of short detection time window due to the rapid metabolism of these drugs in body. This study aimed to investigate the characteristic changes of metabolites in the blood of rats after ingesting diazepam/clonazepam through a gas chromatography-mass spectrometry-based metabolomics method, allowing the indirect reveal of the rats ingested diazepam/clonazepam. First, we found that diazepam and clonazepam in the blood of rats could not be detected by liquid chromatography-tandem mass spectrometry after 48 h of ingestion. Then, orthogonal partial least squares discrimination analysis regression models were, respectively, constructed to determine whether the rats ingested diazepam/clonazepam after 48 h. The results showed that 5 metabolites were found to be associated with diazepam exposure, and 7 metabolites were found to be associated with clonazepam exposure, which may be characterization for the evaluation of digestion of diazepam and clonazepam in rat.