高效液相色谱法测定拉米夫定原料药中对映异构体的含量

HPLC determination of enantiomer content in lamivudine

目的建立测定拉米夫定原料药中对映异构体的HPLC法。方法以InfinityLab Poroshell 120 Chiral-CD(4.6 mm×250 mm,2.7μm)为色谱柱,0.1 mol·L-1醋酸铵-甲醇(70∶30)(A)-甲醇(B)为流动相,梯度洗脱,检测波长270 nm,流速0.6 mL·min-1,柱温25℃,进样体积5μL。结果拉米夫定与对映异构体之间的分离度大于5.0,且完全实现了基线分离,而国内外药典方法均无法实现基线分离。拉米夫定浓度在0.103~1.2961μg·mL-1与峰面积线性关系良好。拉米夫定对映异构体浓度在0.0966~1.2076μg·mL-1与峰面积线性关系良好。结论所建立方法准确、简便、稳定,具有明显的分离效能,可作为拉米夫定中对映异构体的检测方法。

Objective To determine enantiomer content in lamivudine by HPLC.Methods The chromatographic separation was performed on InfinityLab Poroshell 120 Chiral-CD (4.6 mm×250 mm,2.7 μm).The mobile phase was consisted of 0.1 mol·L-1 ammonium acetate-methanol solution (70∶30) (A) and methanol (B).Gradient elution was used with the wavelength at 270 nm.The flow rate was 0.6 mL·min-1,the column temperature was 25℃,and the injection volume was 5 μL.Results The resolution between lamivudine and enantiomer was greater than 5.0,and the baseline separation was completely realized.While the baseline separation was not achieved by domestic and overseas pharmacopoeia standards.The linear relation between the concentration of lamivudine and the peak area at 0.103 and 1.2961 μg·mL-1 was good.The linear relation between the concentration of lamivudine enantiomer and the peak area at 0.0966 and 1.2076 μg·mL-1 was good.Conclusion The method is simple,accurate,and stable,which can obviously separate different components and determine the enantiomer in lamivudine.