LC—MS／MS法同时测定奈韦拉平、拉米夫定、司他夫定、齐多夫定和依非韦伦的血浆浓度

LC-MS/MS method for simultaneous quantification of nevirapine, lamivudine, stavudine, Zidovudine, Efavirenz in human plasma

目的建立高效液相色谱串联质谱技术（LC—MS／MS）同时检测艾滋病常用的5种药物：奈韦拉平（NVP）、拉米夫定（3TC）、司他夫定（d4T）、齐多夫定（AZT）和依非韦伦（EFV）的血药浓度方法。方法采用蛋白沉淀法提取，Eclipse XDB—C18色谱柱（5μm，4．6mm×150mm）分离药物，以（甲醇＋0.3％甲酸）：（水＋0．3％甲酸）（V：V）=80．20为流动相，流速为0．5mL／min，采用质谱多反应监测方法（MRM）检测。结果LC—MS／MS对AZT，3TC的线性范围为25～3200μg/L，NVP和EFV的线性范围为50-6400μg／L，最低检测限均为1μg／L，d4T的线性范围为100～3200μg／L，最低检测限为5μg/L，r均大于0．99。高、中和低3个浓度的日内精密度小于10％，日间精密度小于15％，绝对回收率均大于50％。结论采用HPLC—MS／MS法能同时怯谅、准确测定血浆中这常用的百种药物．

Objective To develop a liquid chromatography/tandem mass spectrometry （LC-MS/MS） method to simultaneously quantify nevirapine （NVP）, lamivudine （3TC）, stavudine （d4T）, Zidovudine （AZT） and Efavirenz （EFV） in human plasma. Methods After protein precipitation, the analytes were chromatographed on a Eclipse XDB-C18 （150 mm×4.6mm i.d., 5μm particle size） column using 20μL injection volume with a run time of 8.0 min. An isocratic mobile phase consisting of 0.3% formic acid in water： 0.3% formic acid in methanol （20：80, v/v） was used to separate these five drugs. The precursor and product ions of these drugs were monitored on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring mode （MRM） without polarity switch. Results The calibration curves of each drug showed a good linearity in a range of concentration between 25-3200μg/L for 3TC, AZT, 50-6400μg/L for EFV, NVP, 50-3200μg/L tor d4T. The relative recoveries for analysts （above 50%） achieved from spiked plasma samples were consistent and reproducible. Intra and inter precision （RSD%） were less than 10% and 15% respectively. Conclusions HPLC-MS/MS is a rapid and precise approach to determine these five AIDS drugs in clinical settings.