替米沙坦血药浓度 HPLC 测定方法的研究

Study on the Plasma Level Determination of Telmisartan by HPLC

目的:建立替米沙坦血药浓度的高效液相-荧光色谱分析方法。方法:以依贝沙坦为内标,血浆样品用乙腈沉淀蛋白,色谱柱:Inersil ODS C_(18)色谱柱,(5μm,150mm×4.6mm),在线过滤器;流动相为水相(0.25%三乙胺水溶液)-乙腈(40∶60,v/v),20%磷酸调节pH为5.0;λ_(ex)=255nm,λ_(em)=390nm;流速1.0mL·min^(-1),柱温为25℃。结果:替米沙坦在6.25-3200ng·mL^(-1)的范围内有良好的线性关系(r=0.9998),最低检测浓度为1.25ng·mL^(-1)(S/N>3),该方法的相对回收率为99.7%-102.3%(n=5);绝对回收率为96.7%-105.3%(n=5);日内 RSD 为2.0%-5.9%,日间 RSD 为1.1%-6.2%。结论:本方法简便,准确,灵敏,特异性强,重现性好,可用于血浆中替米沙坦的测定及人体内药代动力学研究。

Objective:To establish HPLC-fluorescence method for determination of telmisartan in human plasma. Methods:The plasma protein was precipitated by aeetonitrile.The resulting supernatant liquid was directly injected after centrifuge.The Inersil column(ODS C_(18),5μm,150mm×4.6mm)was used as analytical column with a mo- bile phase consisted of 0.25% triethylamine-acetonitrile(40:60,pH was adjusted to 5.0 with phosphoric acid). The flow rate was 1.0 mL·min^(-1)with fluorescence detection set at λ_(ex) 255 nm and λ_(em)390 nm.Irbesartan was a- dopted as the internal standard.Results:The linear range of calibration curve was within drug plasma concentrations of 6.25-3200ng·mL^(-1)(r=0.9998),the detection limit concentration was 1.25ng·mL^(-1)(S/N >3),the abso- lute recovery was 96.7%-105.3%(n=5),the method recovery was 99.7%-102.3%(n=5),the within-day RSD and between-day RSD were 2.0%-5.9% and 1.1%-6.2%(n=5).Conclusions:The method was shown to be sensitive,accurate,simple and good reproducibility for the determination of telmisartan levels in human plas- ma.It was suitable for pharmacokinetics study of telmisartan.