CAJ | 학술논문

目的：建立测定大鼠肝微粒体中赛米司酮的RP-HPLC分析方法并探究其酶促动力学特征。方法 Inertsil ODS-3（150×4.6mm ，5μm ）色谱柱，有机相：水相＝60∶40，有机相为甲醇：乙腈＝3∶2，水相为0.01M KH2 PO4（pH 2.73），检测波长305nm。根据Lineweaver-Burk方程分析数据，求算酶促动力学参数 Km、Vmax及肝脏内在清除率CLint （Vmax/Km）。结果赛米司酮浓度在24.87μg/L～99.48 mg/L浓度范围内线性关系良好（R2＝0.9998），方法回收率在94.7～105.35％之间，日内和日间RSD均小于10％，赛米司酮代谢酶促动力学参数 Km＝10.273μmol/L ，Vmax＝0.3226μmol /（L·min·mg protein），CLint＝31.4μL/（min· mg protein ）。结论此分析方法符合方法学验证要求且简单准确，可满足赛米司酮肝微粒体代谢的研究。
목적：건립측정대서간미립체중새미사동적RP-HPLC분석방법병탐구기매촉동역학특정。방법 Inertsil ODS-3（150×4.6mm ，5μm ）색보주，유궤상：수상＝60∶40，유궤상위갑순：을정＝3∶2，수상위0.01M KH2 PO4（pH 2.73），검측파장305nm。근거Lineweaver-Burk방정분석수거，구산매촉동역학삼수 Km、Vmax급간장내재청제솔CLint （Vmax/Km）。결과새미사동농도재24.87μg/L～99.48 mg/L농도범위내선성관계량호（R2＝0.9998），방법회수솔재94.7～105.35％지간，일내화일간RSD균소우10％，새미사동대사매촉동역학삼수 Km＝10.273μmol/L ，Vmax＝0.3226μmol /（L·min·mg protein），CLint＝31.4μL/（min· mg protein ）。결론차분석방법부합방법학험증요구차간단준학，가만족새미사동간미립체대사적연구。
Objective To establish a RP-HPLC method for the determination of cymipristone and study its enzymatic kinetics in rat liver microsomes .Methods The analysis was performed on Inertsil ODS -3 analytical column(150 × 4.6mm ,5μm) .The mobile phase consist of 60% organic phase and 40% phosphate buffer (pH2.73) .The detection wavelength was 305nm .Lineweaver-Burk equation was employed to determining the enzyme kinetic parameters of cymipristone in vitro metabolic system .The kinetic parameters Km ,Vmax and liver Intrinsic clearance rate CLint(Vmax/Km)were calculated .Results The standard curve was linear in the range of 24.87μg/L-99.48 mg/L (R2=0.9998) .The methods recovery was between 94.7-105.35% .And the intra-day and inter-day precisions(RSD) were less than 10% .In the metabolic system , the enzyme kinetics parameters of cymipristone were found as follows :Km = 10.273μmol/L , Vmax =322.58nmol/L/(min·mg protein) and CLint =31.4μL/(min·mg protein) ,respectively .Conclusion The RP-HPLC method can meet the requirement of methodological verification ,and it is simple ,accurate and can be used for the in vitro metabolism research of cymipristone .