HPLC手性固定相法拆分恩替卡韦光学异构体

Enantio-separation of entecavir opticalisomers by HPLC on Chiralpak AD-H column

建立恩替卡韦5个光学异构体的高效液相色谱拆分方法。采用Chiralpak AD-H手性柱(250 mm×4.6 mm,5μm)在正相条件下进行同时拆分,流动相为正己烷-异丙醇-乙醇-二乙胺(70∶6∶24∶0.1),流速为0.5 mL/min,检测波长为254 nm,柱温为30℃。色谱条件的优化过程中,考察了流动相组成及比例、酸碱添加剂、流速、柱温等因素对恩替卡韦异构体分离的影响。在所建立的条件下,恩替卡韦5个光学异构体分离良好,分离度依次为1.923,4.498,4.573和2.143。该方法简单快速、重复性好,适用于恩替卡韦原料药的光学异构体检查。

A chiral HPLC method was established for the separation and determination of entecavir opticalisomers.Chiral separation of 5 entecavir opticalisomers was achieved under normal-phase chromatographic mode by the Chiralpak AD-H chiral stationary phase.The optimized chromatographic conditions were a mobile phase consisting of n-hexane-isopropanol-alcohol-diethylamine（70 ∶6 ∶24 ∶0.1） with a flow rate of 0.5 mL/min,and an UV detector at 254 nm.The column temperature was set at 30 °C.The influences of the kinds and proportion of organic solvents,acid and base addictives,flow rate and column temperature on the enantio-separation were studied.Under the chromatographic conditions,5 entecavir opticalisomers can be completely separated from each other and the resolution values were not less than 1.9.The validated method is suitable for the quality control of entecavir.