LC-MS/MS法测定人血浆中拉米夫定的质量浓度及生物等效性评价

Determination of lamivudine in human plasma by LC-MS/MS and its application to a bioequivalence study

目的建立人血浆中拉米夫定质量浓度的LC-MS/MS测定方法,并应用于拉米夫定片在健康人体内的生物等效性研究及评价。方法前处理方法采用沉淀蛋白法。分离选用Venusil ABS C18色谱柱,以甲醇-水-甲酸（体积比5.0∶95.0∶0.1）为流动相,采用多反应离子监测（multiple reaction monitoring,MRM）模式进行正离子检测。结果拉米夫定的线性范围为0.020 0～5.00 mg·L-1,定量下限为20μg·L-1。方法准确度为95.0%～102.4%,日内和日间精密度均不大于7.6%。结论本方法适用于拉米夫定人体生物等效性试验研究。24名健康受试者口服拉米夫定参比制剂和受试制剂后的药代动力学行为一致。

Objective To develop a sensitive and rapid LC-MS/MS method to determine the concentration of lamivudine in plasma of healthy human and investigate the bioequivalence of lamivudine tablets in Chinese healthy volunteers. Methods After treated with the method of protein precipitation, the plasma samples were separated on a Venusil ABS C18 column and detected with a triple quadrupole tandem mass spectrometer in positive electrospray ionization mode. Methanol- water- formic acid （ V： V： V = 5.0： 95.0： 0. 1 ） was used as the isocratic mobile phase. Results A linear relationship was established over the concentration range of 0. 020 0 - 5.00 mg. L - 1. The lowest limit of quantitation of lamivudine was 20 p,g. L - 1. The accuracy was in the range of 95.0% - 102. 4% and the Intra-day and inter-day precision was lower than 7.6% in terms of relative standard deviation （RSD）. Conclusions A rapid, simple and sensitive LC-MS/MS method with short analytical time（ 1.6 min each sample） was successfully developed and the method could be applied to the bioequivalence study of lamivudine tablet. According to the results calculated by DAS 3.2.2 software, there is no significant difference in the main pharmacokinetic parameters of 24 healthy male Chinese volunteers af- ter an oral administration of 300 mg lamivudine test and reference formulations.