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以美托洛尔为内标,建立了HPLC-UV测定大鼠血浆、尿、粪、胆汁及肝脑组织匀浆中药物浓度的方法。流动相为甲醇、乙腈、水及0.2mol·L-1醋酸钠缓冲液,以C18柱分离,紫外检测波长232nm,生物样品在不同条件下经两次乙醚处理可获得良好分离。血浆样品最低检测浓度为5ng·ml-1,日内RSD和日间RSD为3.17~10.88%,平均回收率为79.35~95.72%。本法简便、灵敏、可靠,经大鼠单次灌胃20mg·kg-1后血药浓度的测定验证,可满足临床前药代动力学研究的需要。
Metoprolol was used as an internal standard to establish a method for the determination of drug concentration in plasma, urine, feces, bile and liver and brain homogenates by HPLC-UV. The mobile phase consisted of methanol, acetonitrile, water and 0.2mol·L-1 sodium acetate buffer. The mobile phase was separated on a C18 column and detected by UV at a wavelength of 232nm. The biological samples were well separated by two ether treatments under different conditions. The minimum detectable concentration of plasma samples was 5 ng · ml-1. The intra-day RSD and daytime RSD were 3.17-10.88%. The average recovery was 79.35-95.72%. The method is simple, sensitive and reliable. It is verified by the determination of plasma concentration after a single intragastric administration of 20 mg · kg-1 in rats to meet the needs of preclinical pharmacokinetic studies.