反相高效液相色谱法测定人血浆中拉米夫定浓度

Determination the Content of Lamivudine in Human Plasma by RP-HPLC

目的:建立RP-HPLC测定人体血浆中拉米夫定浓度。方法:采用ODS色谱柱(4．6 mm×150 mm,5μm),以乙腈: 40 mmol·L-1的KH2PO4缓冲液(含0．1％三乙胺,pH 3．5)=18:82为流动相,法莫替丁为内标;流速:1．0 ml·min-1;紫外检测波长为270 nm。结果:拉米夫定在2-2 500 ng·ml-1。浓度范围内线性关系良好(r=0．997 3),最低检测限为0．5 ng·ml-1,平均绝对回收率为91．6％(3．2％,n=15),日内与日间RSD均小于5％。结论:本法操作简单,灵敏度高,可以用于拉米夫定的药动学研究及血药浓度监测。

Objective： A new high-performance liquid chromatography （HPLC） assay was developed for the determination the content of lamivudine in human plasma. Method： A reversed phase C18 column was used for the separation of lamivudine and famotidine （ internal standard）, and mobile phase was acetonitrile - 40 mM potassium dihydrogen phosphate solutions containing 0. 1% Triethylamine （ pH 3.5 ） （ 18： 82, v/v） . The detector was operated at the 270 nm wavelength. The flow rate was 1.0ml·min^-1 and the injected volume was 50μl. Plasma samples were processed by liquid-liquid extraction using diehloromethane-isopropyl alcohol （1： 1, v/v） and followed by concentrated. Result： No interference in the assay from any endogenous substance was observed. The calibration curve for lamivudine assay exhibited excellent linear relationship with a correlation coefficient 0.9973 in the concentration range of 2.0-2 500 ng·ml^-1. The absolute recovery of assay was 91.6% （ 3.2% , n = 15 ）. The within - day and inter - day precision （RSD） was less than 5%. The limit of detection was 1.0 ng·ml^-1. Conclusion： The method is simple, accurate and allows to be used as analytical method to analyze lamivudine samples from clinical studies.