HPLC法测定门冬氨酸钾镁注射液中门冬氨酸有关物质及含量

目的：建立门冬氨酸钾镁注射液中门冬氨酸有关物质含量测定的HPLC方法。方法：采用SPHERISORB NH₂柱（4.6 mm×250 mm，5μm）；流动相为乙腈-0.05 mol·L^-1的磷酸二氢钾（60：40）；流速为1.0 mL·min^-1；检测波长为210 nm；柱温为30℃，进样量为5μL；对最大单个杂质采用在线二维脱盐HPLC与MS联用进行初步推断。结果：在该选定色谱条件下，主峰与有关物质峰分离度良好；门冬氨酸在0.338 64~3.386 4 mg·mL^-1浓度范围内线性关系良好（r=0.999 8）；平均回收率为99.6%（n=9），检测限为23.5 ng；初步推断最大单个杂质为富马酸。结论：本方法专属、灵敏、简便，可用于测定门冬氨酸钾镁注射液中门冬氨酸含量和有关物质。

Objective: To establish an HPLC method for the determination of aspartate in potassium aspartate and magnesium aspartate injection and its related substances.Methods: The separation was performed on a Waters SPHERISORB NH₂(4.6 mm×250 mm,5μm)column with the mobile phase consisting of acetonitrile-0.05 mol·L^-1 phosphate solution(60:40) at the flow rate of 1.0 mL·min^-1.The detection wavelength was set at 210 nm.The column temperature was 30℃.The volume of injection was 5 μL.On-line two-dimensional desalting HPLC/MS was applied to infer the maximum single impurity preliminarily.Results: Under above conditions,the main peak and the peaks of the related substances were well separated.Aspartate had a good linear relationship (r=0.999 8) within the range of 0.338 64-3.386 4 mg·mL^-1.The average recovery (n=9) was 99.6%.The limit of the detection was 23.5 ng.The maximum single impurity in related substances was preliminarily inferred as fumaric acid.Conclusion: The method is sensitive,specific and simple.It can be used for determining aspartate in potassium aspartate and magnesium aspartate injection and its related substances.