HPLC法测定替诺福韦的血药浓度及其临床应用

Determination of tenofovir blood drug concentration by HPLC and its clinical application

目的建立测定人血浆中替诺福韦含量的HPLC法。方法血浆样品用蛋白沉淀法处理后,氮气吹干,流动相复溶。色谱条件:色谱柱Ultimate?XB C18(150 mm×4.6 mm,5μm),流动相为15 mmol·L-1四丁基氢氧化铵(p H 7.2)-乙腈(78∶22),流速1 m L·min-1,柱温35℃,紫外检测器,检测波长262 nm。外标法定量。结果在该色谱条件下,替诺福韦与内源性物质及同时服用药物完全分离,最低定量质量浓度为20μg·L-1,在20~1 000μg·L-1内线性关系良好(r=0.9986)。结论本方法简便快速,专属性强,灵敏度高,可适用于临床患者替诺福韦的血药浓度监测。

AIM To develop a high performance liquid chromatography-ultraviolet( HPLC-UV) method to determine tenofovir in human plasma. METHODS Tenofovir in plasma was precipitated by acetonitrile,dried by nitrogen and dissolved with mobile phase. The HPLC separation was carried out on an Ultimate?XB C18column( 150 mm × 4. 6 mm,5 μm) with mobile phase of 15 mmol·L-1etrabutylammonium hydroxide solution-acetonitrile( 78∶ 22) at a flow rate of 1 m L·min-1. Column temperature was 35 ℃. Tenofovir was detected at wavelength of 262 nm. RESULTS The good linearity of tenofovir under chromatography was shown. Detection limits of tenofovir ranged from 20-1 000 μg·L-1( r = 0. 998 6). CONCLUSION HPLC-UV is sensitive and specific for tenofovir analysis in human plasma and can be applied to clinical patients with tenofovir blood drug concentration monitoring.