反相高效液相色谱法同时检测4种细胞色素P450探针药物  

Determination of Four Cytochrome P450 Probe Drugs by RP-HPLC in a Single Run

作　　者：胡道德[1] 顾磊[1] 姚慧娟[1] 刘文[1] 王松坡[1] 刘皋林[1] 

机构地区：[1]上海交通大学附属第一人民医院临床药学研究室,上海200080

出　　处：《中国药学杂志》2010年第1期71-74,共4页Chinese Pharmaceutical Journal

基　　金：上海中医药科研基金资助项目(2004J009C)

摘　　要：目的建立同时检测小鼠血浆中4种细胞色素P450（CYP450）探针药物,即咖啡因（CYP1A2）、氨苯砜（CYP3A4）、苯妥英（CYP2C9）和氯唑沙宗（CYP2E1）的反相高效液相色谱方法。方法血浆样品用乙酸乙酯提取处理。色谱柱为Dikma Dia-monsil-C18（4.6 mm×200 mm,5.0μm）,保护柱4.6 mm×10 mm（Agilent）,流动相为甲醇-磷酸盐缓冲系（含0.02 mol.L-1的磷酸二氢钾,0.02 mol.L-1的三乙胺,磷酸调pH6.5）,体积比为48∶52,流速1.0 mL.min-1,柱温25℃,安替比林为内标,检测波长为265 nm。结果咖啡因、氨苯砜、苯妥英和氯唑沙宗的线性范围分别为0.2~1.6,0.075~0.600,0.2~1.6,0.25~2.00 mg.L-1,4种探针药物日内、日间RSD均〈5.14%,方法回收率均〉95.0%（RSD〈6.14%）。结论该反相高效液相色谱法快速、准确、灵敏度高、专属性强,可用于探针药物评价主要CYP450亚型酶的活性。OBJECTIVE To develop a reversed-phase high performance liquid chromatographic（RP-HPLC） method for the determination of four cytochrome P450 in rat plasma.METHODS The four specific probe substrates of caffeine,dapsone,phenytoin and chlorzoxazone,together with the internal standard antipyrine,were extracted using liquid liquid extraction in rat plasma,followed by RP-HPLC using a Dikma Diamonsil-C18（4.6 mm×200 mm,5.0 μm）.The mobile phase consisted of a methanol and 0.02 mol·L-1 potassium dihydrogephosphate buffer solution（48∶52,adjusted to pH 6.5 with phosphate）.All analytes were separated simultaneously in a single run.RESULTS The linear concentration ranges of the calibration curves for caffeine,dapsone,phenytoin and chlorzoxazone were 0.2-1.6,0.075-0.600,0.2-1.6 and 0.25-2.00 mg·L-1,respectively.The intra-day and inter-day relative standard deviations（RSD） were less than 5.14%.The methodological recoveries of the four probes were more than 95.0%（RSD6.14%）.CONCLUSION The RP-HPLC method is rapid,sensitive,precise and selective,and can be suitable for determining the plasma concentration of these probes and evaluating the CYP1A2,CYP3A4,CYP2C9 and CYP2E1 activities in the rats.

关 键 词：反相高效液相色谱法 Cocktail探针药物 咖啡因 氨苯砜 苯妥英 氯唑沙宗 淫羊藿提取物 细胞色素P450 

分 类 号：R917[医药卫生—药物分析学]