Zhai Xuejia, Lu Yongning
Department of Pharmacy, Union Hospital, Huazhong University of Science and Technology, Wuhan, China.
Pharmazie. 2013 Jan;68(1):19-26.
Cytochrome P450 enzymes are responsible for the oxidative metabolism of most pharmaceutical compounds. A "cocktail" approach which employs simultaneous administration of a mixture of substrates of CYP enzymes was often used to assess the metabolic activity of multiple P450 forms in one experiment. Phenacetin, coumarin, tolbutamide, chlorzoxazone and testosterone are commonly used as probe substrates to evaluate cytochrome P450 function. An analytical strategy to simultaneously extract and analyze the five probe substrates and their major metabolites by HPLC-DAD was developed. The incubation was done with all the substrates in one step. The ten analytes were extracted simultaneously by solid-phase extraction (SPE) from rat liver microsomes. A C18 analytical column and mobile phase composed of acetonitrile and 0.02% aqueous phosphoric acid were used for the chromatographic separation with DAD detection. Limits of quantification varied between 0.02378 and 0.2361 microg/mL which contributed to quantify all these drugs and metabolites with UV detection. The method is applicable for the modeling and description of pharmacological interactions on rat cytochromes P450 or can be used for in vitro evaluation of cytochromes 1A2, 2A6, 2C11, 2E1 and 3A2.
细胞色素P450酶负责大多数药物化合物的氧化代谢。一种“鸡尾酒”方法，即同时给予CYP酶底物混合物，常用于在一个实验中评估多种P450形式的代谢活性。非那西丁、香豆素、甲苯磺丁脲、氯唑沙宗和睾酮通常用作探针底物来评估细胞色素P450的功能。开发了一种通过HPLC-DAD同时提取和分析五种探针底物及其主要代谢物的分析策略。所有底物在一步中进行孵育。通过固相萃取（SPE）从大鼠肝微粒体中同时提取十种分析物。使用C18分析柱和由乙腈和0.02%磷酸水溶液组成的流动相进行色谱分离，并采用DAD检测。定量限在0.02378至0.2361μg/mL之间，这有助于通过紫外检测对所有这些药物和代谢物进行定量。该方法适用于大鼠细胞色素P450上药理相互作用的建模和描述，或可用于细胞色素1A2、2A6、2C11、2E1和3A2的体外评估。
