Determination of brain cytochrome P450 2E1 activity in rat with the probe of chlorzoxazone by liquid chromatography–mass spectrometry

A simple and sensitive method was developed for the determination of cytochrome P450 2E1 (CYP2E1) activity based on the liquid chromatography–mass spectrometry (LC–MS) analysis of 6-hydroxychlorzoxazone generated by 6-hydroxylation of chlorzoxazone under specific catalysis of CYP2E1. In the proposed method, 2-benzoxazolinone was chosen as internal standard and isopropyl ether was used as extraction solvent for sample preparation. The inter-day and intra-day precisions at low, medium and high concentrations of 6-hydroxychlorzoxazone were below 20.0%, and the LOD (S/N = 3) was 0.05 ng/mL. This method was applied to analyze the CYP2E1 activity of rat in different brain regions including frontal cortex (FC), cerebellum (CB), brain stem (BS), hippocampus (HC), striatum (ST), thalamus (TH), and olfactory bulb (OB). The results confirmed that chlorzoxazone was a suitable probe for the determination of CYP2E1 activity in brain regions and samples with low content of CYP2E1.