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目的:建立HPLC-MS法测定大鼠血浆中辛伐他汀及其代谢物辛伐他汀酸的浓度。方法:血浆样本加入适量内标和醋酸铵缓冲液,以甲基叔丁基醚萃取后采用LC-MS进行分析。色谱柱采用Inertsil ODS-3柱(150 mm×2.1 mm,5.0μm);流动相由乙腈-2.5 mmol.L-1醋酸铵(含0.1%甲酸)(75∶25)组成,柱温35°C;流速0.3 mL.min-1;采用电喷雾离子源(ESI),以多反应监测方式(MRM)进行定量分析。辛伐他汀和内标洛伐他汀在正离子模式下定量分析离子对分别为m/z 419.2→m/z199.2和m/z 405.2→m/z 199.2;辛伐他汀酸和内标洛伐他汀酸在负离子模式下定量分析离子对分别为m/z 435.2→m/z319.2和m/z 421.4→m/z 319.2。结果:辛伐他汀和辛伐他汀酸在5.0～6 400 ng.mL-1内线性关系良好(r>0.999),最低定量限为0.1 ng.mL-1,提取回收率为87.91%～99.77%,日内、日间精密度均不高于8.95%。结论:该方法分析速度快、灵敏、准确,为临床进一步研究辛伐他汀提供了基础。
OBJECTIVE: To establish a HPLC-MS method for the determination of simvastatin and its metabolites simvastatin acid in rat plasma. Methods: Plasma samples were spiked with appropriate amount of internal standard and ammonium acetate buffer and extracted with methyl t-butyl ether and analyzed by LC-MS. The column was equipped with an Inertsil ODS-3 column (150 mm × 2.1 mm, 5.0 μm). The mobile phase consisted of acetonitrile-2.5 mmol.L-1 ammonium acetate (0.1% formic acid) ; Flow rate of 0.3 mL.min-1; using electrospray ionization source (ESI), quantitative analysis by multiple reaction monitoring (MRM). Simvastatin and internal standard lovastatin were quantified in positive ion mode by ion pair m / z 419.2 → m / z 199.2 and m / z 405.2 → m / z 199.2 respectively; simvastatin acid and internal standard log The statin ion quantification ion pair in negative mode was m / z 435.2 → m / z 319.2 and m / z 421.4 → m / z 319.2, respectively. Results: The linear relationship between simvastatin and simvastatin acid was good (r> 0.999) in the range of 5.0-6 400 ng.mL-1, the lowest limit of quantification was 0.1 ng.mL-1, the recovery was 87.91% -99.77% , Day, day precision not higher than 8.95%. Conclusion: The method is rapid, sensitive and accurate and provides the basis for further study of simvastatin.