HPLC法同时测定肝微粒体中右美沙芬和咪达唑仑的代谢速率

Simultaneous determination of dextromethorphan and midazolam in rat liver microsome by HPLC

目的:建立大鼠肝微粒体中右美沙芬和咪达唑仑测定的高效液相色谱法,并考察其在大鼠肝微粒体内的代谢速率.方法:以地西泮为内标,样品在肝微粒内孵育,用乙腈沉淀蛋白浓集后进样分析.流动相:乙腈-磷酸盐缓冲液(0.01mol L-1 K2HPO4-0.5%三乙胺,HCl调PH至7.0)=40:60;检测波长:202 nm;柱温:30℃.结果:该方法能够满足肝微粒体内右美沙芬和咪达唑仑的含量测定要求.DM和MD的代谢速率分别是0.4406±0.066 nmol mg-1.min-1和0.7582±0.018 nmol mg-1.min-1.结论:本方法简单、专属性好、准确度高、灵敏度好,可用于肝微粒中右美沙芬和咪达唑仑的代谢研究.

Objective：To develop an HPLC method for the determination of dextromethorphan and midazolam,and investigate their metabolic velocity in rat liver microsome.Method：After being incubated in liver microsome,dextromethorphan and midazolam are extracted with acetonitrile.The organic solution is evaporated and concentrated by mobile phase for analysis.The chromatographic conditions are adopted as follows：mobile phase is acetonitrile-phosphate buffer（0.01 moloL-1 K2HPO4-0.5% triethylamine,add HCl to adjust the pH value of 7.0） = 40：60;detection wavelength is 202 nm;column temperature is 30 ℃.Result：This method is fully validated and the metabolic velocity of dextromethorphan and midazolam are 0.4406±0.066 nmolomg-1·min-1 and 0.7582±0.018 nmolomg-1·min-1.Conclusion：The method is simple,specifical,accurate,precise and sensitive.The successful application demonstrates that the developed method could be readily used in the study of the metabolism of dextromethorphan and midazolam in liver microsome.