高效液相色谱法测定苯甲酸阿格列汀片中对映异构体的含量

High-Performance Liquid Chromatography for the Separation of Alogliptin Benzoate and Its Enantiomer

关键词:  高效液相色谱法  手性分离  苯甲酸阿格列汀  药物质量控制

Key words:  high-performance liquid chromatography  chiral separation  alogliptin benzoate  drug quality control

摘要:

目的 建立高效液相色谱方法分离苯甲酸阿格列汀的对映异构体，并对异构体含量进行测定。方法 使用以硅胶表面涂敷直链淀粉-三（3,5-二甲基苯基氨基甲酸酯）为填充剂的色谱柱Daicel CHIRALPAK AD-H（4.6mm×250mm，5μm）；检测波长为278 nm；柱温为40℃；流速为1.0 mL/min；流动相比例为：正己烷：无水乙醇：甲烷磺酸（80：20：0.3）；进样体积为20 μL。结果 苯甲酸阿格列汀及其对映异构体在优化的手性分离条件下能够被完全分离；出峰顺序为对映异构体峰在前，阿格列汀峰在后；在0.02 ~ 2 μg·mL-1范围内，苯甲酸阿格列汀对映异构体的质量浓度与峰面积呈现良好的线性关系；苯甲酸阿格列汀及其对映异构体的加标回收率在98.90% ~ 101.24%之间，相对标准偏差为0.97%，表明该法准确度良好。结论 建立的方法简便、准确、灵敏度高可用于苯甲酸阿格列汀及其对映异构体的分离及定量。

Abstract:

To establish a high-performance liquid chromatography (HPLC) method for the separation of enantiomer of alogliptin benzoate and to determine the content of the enantiomer. METHODS A chiral column packed with cellulose tris(3,5-dimethylphenylcarbamate) coated on silica gel, Daicel CHIRALPAK AD-H (4.6 mm × 250 mm, 5 μm), was used. Detection wavelength was set at 278 nm; column temperature was maintained at 40°C; flow rate was 1.0 mL/min; the mobile phase consisted of n-hexane: anhydrous ethanol: methanesulfonic acid in the ratio of 80:20:0.3; and the injection volume was 20 μL. RESULTS Under the optimized chiral separation conditions, alogliptin benzoate and its enantiomer were completely separated, with the enantiomer peak appearing before the alogliptin peak. A good linear relationship was observed between the mass concentration of alogliptin benzoate enantiomer and the peak area in the range of 0.02 to 2 μg·mL-1. The recovery rates of alogliptin benzoate and its enantiomer spiked were between 98.90% ~ 101.24%, with relative standard deviations of 0.97%, indicating good accuracy of the method. CONCLUSION The established method is simple, accurate, and sensitive, and can be used for the separation and quantification of alogliptin benzoate and its enantiomer.