高效液相色谱法分离测定恩替卡韦光学异构体

摘要

本文建立了一种简单有效的高效液相-紫外检测法测定分离恩替卡韦光学异构体.恩替卡韦化学结构中有3个手性原子,恩替卡韦有7个光学异构体,其中有6个非对映异构体,1个对映异构体.在Symmetrix ODS-AQ C18 (4.6 mm×250 mm,5μm)色谱柱上实现了恩替卡韦与非对映异构体的分离,在Daicel CHIRALPAK AD (4.6 mm×250 mm,10 μm)色谱柱上实现了恩替卡韦和对映异构体的分离.检测波长为254 nm.非对映异构体(非对映异构体-1,非对映异构体-2,非对映异构体-3)的检测限和定量限分别为0.0371,0.0376,0.0377 μg/mL和0.124,0.125,0.126 μg/mL.对映体的检测限和定量限分别为0.14和0.46 μg/mL.非对映异构体-1,非对映异构体-2,非对映异构体-3和对映异构体的精密度分别为0.36％,0.44％,1.04％和0.67％.对映异构体的回收率为98.4％-100.5％.该方法可用于控制恩替卡韦光学异构体的杂质限度.%A simple and effective high-performance liquid chromatography with ultraviolet detection (HPLC-UV) method was established to determine entecavir optical isomers.With 3 chiral carbon atoms,entecavir had 7 optical isomers,including 6 diastereoisomers and 1 enantiomer.The separations were performed on a Symmetrix ODS-AQ C18 column (4.6 mm×250 mm,5 μm) and a Daicel CHIRALPAK AD column (4.6 mm×250 mm,l0 μm),respectively.The detection wavelength was set at 254 nm.The limit of detection (LOD) and the limit of quantification (LOQ) of diastereoisomers (diastereoisomer-l,diastereoisomer-2,diastereoisomer-3) were 0.0371,0.0376,0.0377,and 0.124,0.125,0.126 μg/mL,respectively.The LOD and LOQ of enantiomer were 0.14 and 0.46 lag/mL,respectively.The precision was within 0.36％,0.44％,1.04％,and 0.67％ for diastereoisomer-1,diastereoisomer-2,diastereoisomer-3,and enantiomer,respectively.The recoveries of enantiomers ranged from 98.4％ to 100.5％.The method could be applied to control the quality of entecavir.