手性固定相高效液相色谱法拆分富马酸替诺福韦二吡呋酯对映异构体

摘要

A new chiral high-performance liquid chromatographic method for the enantiomeric separation of tenofovir DF was established.The (R) and (S) isomers were baseline resolved on Chiralcel OD-H column (500min×4.6mm,5μm,Daicel) at 20℃.Satisfactory results were achieved under normal-phase chromatographic mode with the mobile phase containing hexane and ethanol anhydrous in the ratio of 95:5(v/v) with 0.2％ of diethylamine.The method was validated for linearity,repeatability,the limit of detection,the limit of quantification,and robustness.The resolution between the enantiomers was not less than 1.78.The limit of detection and the limit of quantification of S enantiomer were found to be 0.005μg and 0.015μg,respectively.The percentage recovery of S enantiomer was ranged from 98.68 to 109.21 in bulk drug samples of tenofovir DF.The sample solution and mobile phase were found to be stable for at least 72h.The optimized method was successfully used for separation of S enantiomer from tenofovir DF and was proven tobe reproducible and accurate for quantification ofS enantiomer in bulk drugs.%目的 建立了手性固定相高效液相色谱法拆分富马酸替诺福韦二吡呋酯对映异构体的方法.方法 采用高效液相色谱法,色谱柱为Chiralcel OD-H (500mm×4.6mm,5μm,Daicel),以环己烷-无水乙醇(95:5)为流动相(含0.2％的二乙胺),检测温度为20℃.该方法经过线性、重复性、检测限、定量限和方法抗干扰性的验证.结果 富马酸替诺福韦二吡呋酯及异构体在此条件下能够很好地分离,分离度为1.78.异构体的检测限和定量限分别为0.005μg和0.015μg.S异构体在原料药中的回收率为98.68％～109.21％,样品溶液和流动相在72h内稳定.结论 该方法能成功的用于富马酸替诺福韦二吡呋酯及异构体的分离,并且能够重现性好的准确定量原料药中的S异构体.