快速测定人血浆中洛匹那韦/利托那韦浓度的LC-MS/MS方法建立

Development of LC-MS/MS method for rapid quantification of lopinavir/ritonavir(Kaletra)in human plasma

目的建立LC-MS/MS方法快速测定人血浆中洛匹那韦（lopinavir,LPV）/利托那韦（ritonavir,RTV）（克力芝）的含量。方法血浆样品采用蛋白沉淀法处理后,以甲醇-水（0.1%甲酸）为流动相进行梯度洗脱,流速为0.2 ml/min,色谱柱采用Thermo Hypersil GOLD柱（2.1 mm×100 mm,5 mm）,柱温为25℃,采用ESI源以选择反应监测（SRM）方式进行正离子检测,用于定量分析的离子对为LPV（m/z 629.3→155.0）、RTV（m/z 721.3→268.0）,内标MS275（m/z 377.1→359.2）。结果两药在人血浆中浓度在20~1000 ng/ml时,线性关系良好（r〉0.994）。日内、日间精密度,低浓度〈20%,中高浓度〈15%,回收率为86.5%~96.3%。结论该检测方法简单、快速、灵敏、准确,适用于两药的临床血药浓度监测及其药物代谢动力学研究。

Objective To establish a rapid LC-MS/MS method for the simultaneous quantitative determination of lopinavir （LPV） and ritonavir（RTV） in human plasma. Methods Plasma samples were prepared by protein precipitation and separated by a Thermo Hypersil GOLD column （2.1 mm× 100 mm, 5 mm）with the mobile phase consisting of methanol and water （0.1% formic acid ） at a flow rate of 0.2 ml/min. Detection of LPV, RTV and the internal standard（IS） MS 275 was performed using selected reaction monitoring （SRM） of the transitions m/z 629.3→155.0, m/z 721.3→268.0 and m/z 377.1→359.2 in positive ion mode, respectively. Resuits The calibration curve was linear in the range of 20-1000 ng/ml （r〉0.994） for LPV and RTV. The intra and inter-day precision and accuracy values met the set acceptance criteria. Conclusion The method is rapid, sensitive and accurate for the therapeutic drug monitoring of LPV and RTV simultaneously in clinic and pharmacokinetic studies.