硝苯地平探针高效液相色谱法测定大鼠肝微粒体CYP3A酶活性  

Determination of CYP3A Activity in Rat Liver Microsomes by HPLC Using Nifedipine as Probe

作　　者：王芳[1] 李艳丽[1] 郑文[1] 许卉[1] 

机构地区：[1]烟台大学药学院,山东烟台264005

出　　处：《烟台大学学报（自然科学与工程版）》2013年第2期119-122,156,共5页Journal of Yantai University(Natural Science and Engineering Edition)

基　　金：国家自然科学基金资助项目(81102780);烟台大学青年学术骨干教师培养计划资助项目

摘　　要：以硝苯地平为探针药物,研究建立一种大鼠肝微粒体CYP3A酶活性定量测定的高效液相色谱方法,并优化色谱条件和肝微粒体样品预处理方法.在测定条件下,硝苯地平经CYP3A酶代谢转化为氧化硝苯地平,其色谱保留时间7.09min,无内源性物质干扰.方法的线性检测范围1.0～40.0μmol/L,定量下限1.0μmol/L,回收率94.03%～98.88%,日内、日间相对标准偏差均小于7.3%.方法学验证结果表明,该方法快速、灵敏、准确,具有良好的专属性、精密度和稳定性,适于生物样品中氧化硝苯地平的定量测定,可用于CYP3A酶活性评价及酶动力学研究.An HPLC method using nifedipine as probe drug has been developed for the determination of CYP3A activity in rat liver microsomes. Both chromatography parameters and pretreatment program of liver microsomes for HPLC assay are optimized. Under the optimal conditions, nifedipine is rapidly metabolized to oxidized nifedipine with a retention time of 7.09 min and no interference from nifedipine or endogenous matrix compounds. The cali- bration curve is linear with the correlation coefficient of 0. 999 5 in the range of 1.0 - 40. 0 μmol/L. The lower limit of quantification of oxidized nifedipine in liver microsomes is 1.00 μmol/L. With the recovery of 94.03% - 98.88% and the intra-day and inter-day relative standard deviations less than 7.3%, the proposed assay is rapid and sensitive for determining oxidized nifedipine in biosamples and evaluating CYP3A activity in vitro.

关 键 词：CYP3A酶活性 大鼠肝微粒体 硝苯地平 氧化硝苯地平 高效液相色谱法 

分 类 号：R969.1[医药卫生—药理学]