LC-MS／MS同时测定6种CYP450探针药物及快速评价小鼠CYP450同工酶活性  

Phenotyping of six main CYP450 probe drugs in mice by an in vivo cocktail and liquid chromatography-tandem mass spectrometry

作　　者：王丹[1] 刘萍霞[2] 张振清[1] 阮金秀[1] 

机构地区：[1]军事医学科学院毒物药物研究所药物代谢动力学重点实验室,北京100850 [2]解放军庐山疗养院药剂科,江西庐山332900

出　　处：《军事医学科学院院刊》2008年第6期545-549,共5页Bulletin of the Academy of Military Medical Sciences

摘　　要：目的:建立同时测定小鼠全血中6种CYP450探针药物,即咖啡因(CYP1A2)、甲苯磺丁脲(CYP2C)、奥美拉唑(CYP2C)、右美沙芬(CYP2D22)、氯唑沙宗(CYP2E1)和咪达唑仑(CYP3A11)的方法,并采用cocktail法快速评价药物对CYP450同工酶的影响。方法:蛋白沉淀法处理血样,离心后取上清,LC-MS/MS检测。选取地塞米松作为药酶诱导剂,对所建立的模型进行验证。结果:咖啡因、甲苯磺丁脲和氯唑沙宗的线性范围均为20～4 000 ng/ ml,咪达唑仑、奥美拉唑和右美沙芬的线性范围均为1～200 ng/ml。6种探针药物的日内及日间精密度均小于15%,方法回收率均大于90%。地塞米松表现出对小鼠CYP450有诱导作用。结论:本方法准确、灵敏、重现性好,可用于cocktail法快速评价整体动物主要CYP450亚型酶的活性。Objective：To develop a liquid chromatography-tandem mass spectrometry method for simultaneous determination of six major cytochrome P450 probe drugs, caffeine （ CYP1A2）, tolbutamide （ CYP2C）, omeprazole （ CYP2C）, dextromethorphan （ CYP2D22）, chlorzoxazone （ CYP2E1 ） and midazolam （ CYP3All ） in mouse blood. Method： All probe drugs were administered simultaneously and blood samples were handled by precipitation of protein. Following centrifugation, the supernatant was analyzed by LC-MS/MS. Dexamethasone was used as a positive control to validate the model. Results： Linear detection responses were obtained for caffeine, tolbutamide and chlorzoxazone concentrations ranging from 20 - 4 000 ng/ml, for midazolam, omeprazole and dextromethorphan concentrations ranging from 1 - 200 ng/ml. The intra- and inter-day precision was less than 15%, and recovery was above 90%. Dexamethasone was proved to be an inducer according to the model. Conclusion： The method was formally validated and showed good performance in terms of linearity, sensitivity, precision and accuracy, and thus can be applied to cytochrome P450 phenotyping studies by an in vivo cocktail in mice.

关 键 词：COCKTAIL CYP450 LC—MS/MS 咖啡因 甲苯磺丁脲 奥美拉唑 右美沙芬 氯唑沙宗 咪达唑仑 地塞米松 

分 类 号：R969.1[医药卫生—药理学]