UPLC-MS/MS法同时测定人血浆中辛伐他汀和辛伐他汀羟基酸的浓度

目的:建立UPLC-MS/MS法同时测定人血浆中辛伐他汀和辛伐他汀羟基酸的浓度。方法:血浆样品用甲基叔丁基醚提取,离心后取上清液氮气吹干,流动相复溶后进行UPLC-MS/MS测定。色谱柱:BEH C18(2.1 mm×50 mm,1.7μm);流动相:乙腈-0.01 mol.L-1醋酸铵(72∶28);流速:0.15 mL.min-1;柱温:40℃,进样量:8μL。电喷雾离子化(ESI),正离子模式多重反应选择离子检测(MRM),辛伐他汀、辛伐他汀羟基酸及内标洛伐他汀的检测离子对分别为:m/z 419→199,437→303,405→199。结果:血浆样品中辛伐他汀、辛伐他汀羟基酸线性范围分别为0.241～61.76 ng.mL-1(r=0.999,n=5)和0.344～88.16 ng.mL-1(r=0.997,n=5),日内、日间精密度(RSD)均小于15%,方法的平均回收率分别为100.6%和106.0%,血浆基质对血浆中的辛伐他汀和辛伐他汀羟基酸测定无干扰。结论:建立的UPLC-MS/MS法处理简单、灵敏、特异性高,定量准确,为辛伐他汀制剂的临床药代动力学研究提供了简便、准确的分析测定方法。

UPLC-MS/MS determination of simvastatin and simvastatin hydroxy acid in human plasma

Objective:To establish a method for simultaneous determination of simvastatin and simvastatin hydroxy acid in human plasma by UPLC-MS/MS.Methods:The plasma sample was extracted by methyl tert-butyl ether.The resulting supernatant liquid after being centrifuged was dried by nitrogen and was analyzed by the UPLC-MS/MS method after it was diluted quantitatively by mobile phase under following chromatographic conditions:BEH C18(2.1 mm×50 mm,1.7 μm),the mobile phase consisting of acetonitrile-0.01 mol·L-1 ammonium acetate(72∶ 28),with flow rate of 0.15 mL·min-1,column temperature at 40 ℃,and sample volume of 8 μL.The results of positive ion MRM detection of simvastatin and simvastatin hydroxy acidwere m/z 419→199,and m/z 437→303,respectively,using lovastatin(m/z 405→199)as the internal standard.Results:A good linearity of simvastatin was shown in the range of 0.241-61.76 ng·mL-1(r=0.999,n=5);of simvastatin hydroxy acid,0.344-88.16 ng·mL-1(r=0.997,n=5).Simvastatin’s and simvastatin hydroxy acid’s intra-day RSD and inter-day RSD were less than 15%.The average recoveries were 100.6% and 106.0%,respectively.The plasma matrix had no effect on the quantification of simvastatin and simvastatin hydroxy acid.Conclusions:A rapid,sensitive and accurate UPLC-MS/MS method was developed and it was proved to be effective for the determination of simvastatin and simvastatin hydroxy acid in human plasma.