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

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 879; no. 3; pp. 260 - 266
• Main Authors: Wu, Jian-Yuan, Yue, Jiang, Feng, Yu-Qi
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.02.2011; Elsevier
• Subjects: 6-hydroxychlorzoxazone; Analysis; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Brain - enzymology; Brain Chemistry; brain stem; Calibration; catalytic activity; cerebellum; Chlorzoxazone; Chlorzoxazone - analogs & derivatives; Chlorzoxazone - analysis; Chlorzoxazone - chemistry; Chlorzoxazone - pharmacokinetics; Chromatography, Liquid - methods; cortex; cytochrome P-450; Cytochrome P-450 CYP2E1 - metabolism; Cytochrome P450 2E1; enzyme activity; Fundamental and applied biological sciences. Psychology; General pharmacology; hippocampus; Linear Models; liquid chromatography; Liquid chromatography–mass spectrometry; mass spectrometry; Mass Spectrometry - methods; Medical sciences; muscle relaxants; olfactory bulb; Pharmacology. Drug treatments; Rats; Reproducibility of Results; Sensitivity and Specificity; thalamus; Tissue Distribution; 6-hydroxychlorzoxazone; Liquid chromatography–mass spectrometry; Cytochrome P450 2E1; Chlorzoxazone; Benzoxazole derivatives; Rat; Enzyme; Cytochrome P450; Liquid chromatography-mass spectrometry; Rodentia; Central nervous system; HPLC chromatography; Determination; Encephalon; Vertebrata; Mammalia; Animal; Quantitative analysis
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2010.12.013

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.