液质联用检测大鼠血、尿液和胆汁中氯沙坦的含量

Determination of losartan concentrations in rat plasma,urine and bile by liquid chromatography-tandem mass spectrometry

目的:建立LC/MS/MS法测定大鼠血?尿液和胆汁中氯沙坦的浓度,研究氯沙坦在大鼠体内的药物动力学特点。方法:大鼠血浆样品采用沉淀蛋白作为前处理方法,尿液和胆汁采用液-液萃取方法进行前处理,进样检测。色谱柱为Agela Venusil MP-C182.1mm×50mm,粒径5μm;流动相A为0.1%甲酸,流动相B为0.1%甲酸(溶剂为乙腈)。采用梯度洗脱:流动相B在1.5min内从20%到95%,流速0.35ml/min,进样量为10μl,质谱仪检测,离子源为ESI源。结果:氯沙坦在血、尿液和胆汁中的线性范围为1～1000μg/L,线性良好(r>0.99),平均相对回收率>80.0%,日内和日间精密度均小于15%。结论:该检测方法操作方便,结果准确,专属性强,可用于血、尿液和胆汁中氯沙坦药物含量的测定。

Objective： To establish a liquid chromatography-tandem mass spectrometry （LC/MS/MS） method for determination of the losartan concentrations in rat plasma, urine and bile. Methods： The method was based on a protein precipitation procedure for rat plasma and liquid-liquid extraction for rat urine and bile,then the diluted extract was analyzed by LC/MS/MS using turbo ion spray （TIS） positive ion mode. Chromatography was performed on an Agela Venusil MP-C18 column （5 um particle,2.1 mm ×50 mm） by gradient elution. Mobile phase A was 0.1% formic acid in water and mobile phase B was 0.1 % formic acid in aeetonitrile. The mobile phase B increased from 20 % to 95 % within 1.5 min,with the flow rate being 0.35 ml/min and the feeding rate being 10 ul. Electrospray ionization （ESI） ion source was used. Results： The linear range of losartan was 1-1 000 ug/L in the plasma,urine and bile of rats （r〉0.99）. The mean relative recovery rate was higher than 80.0% ;RSD values of intra-day and inter-day were both less than 15 %. Conclusion： The present method is easy to perform, accurate, and specific; it can be used for determination of losartan concentration in rat plasma,urine and bile.