高效液相色谱法同时测定5种抗乙肝药物的研究

Simultaneous Determination of Five Anti-hepatitis B Drugs by High Performance Liquid Chromatography

目的建立高效液相色谱法同时测定五种抗乙肝药物。方法采用水-乙腈-三氟乙酸在Waters Xbridge C 18柱(4.6×250mm,5μm)上进行梯度洗脱,流速为1.0mL·min^-1,检测波长为260nm,柱温35℃。结果5种药物在35min内均能达到基线分离,线性范围良好,相关系数均大于0.999;平均回收率n=3为99.4~101.7%(RSD:0.1~1.7%),精密度和稳定性试验的RSD分别为0.6%~0.8%,0.4%~0.5%,检测限0.1~0.2μg·mL^-1。在恩替卡韦制剂样品中检出假阳性样品,检出物为阿德福韦酯。结论该方法适用于五种抗乙肝药物同时测定,并且能准确筛查出高仿假药。

OBJECTIVE To establish a high performance liquid chromatography(HPLC)method for simultaneous determination of five anti-hepatitis B drugs.METHODS The separation was carried out on Waters Xbridge C 18 column(4.6×250mm,5μm)eluted with water-acetonitrile-trifluoroacetic acid with gradient mode at a flow rate of 1.0mL·min^-1.The detection wavelength was 260nm.The column temperature was set at 35℃.RESULTS Good separation of five drugs was achieved within 35 minutes.Calibration curve of five drugs showed good linear relationship(r>0.999,n=6).The mean recoveries ranged from 97 to 107%(n=3,RSD:0.2~2.0%).The RSD of precision were in the range of 0.5%to 1.5%,while those for stability were in the range of 0.8%to 2.0%.Meantime the LOD were in the range of 0.1-0.2μg·mL^-1.A counterfeit drug was detected in the entecavir formulation sample which was adefovir dipivoxil.CONCLUSION This method can be used for determination of five anti-hepatitis B drugs,and can accurately screen high-fake drugs.