HPLC法测定门冬氨酸鸟氨酸原料药中两种异构体的含量

Determiantion of the isomers in L-ornithine-L-aspartate API by HPLC

目的采用高效液相色谱法对门冬氨酸鸟氨酸原料药中的两种异构体进行测定。方法色谱柱为Chirex3126(D)-Penicillamine(250 mm×4.6 mm,5μm)柱,以浓度1.5 mmol·L^(-1)硫酸铜和80mmol·L^(-1)醋酸铵的混合溶液为流动相,流速为0.6 m L·min^(-1),柱温为20℃,检测波长为280 nm。结果门冬氨酸和鸟氨酸与异构体能有效分离,分离度大于1.5,其他杂质不干扰测定。D-鸟氨酸和D-门冬氨酸分别在质量浓度1.03~19.69、2.06~19.92 mg·L^(-1)内呈良好的线性关系,回收率分别为99.2%和96.9%。结论该方法可以适用于该原料药中异构体的检测,为原料药的质量控制提供更好的依据。

Objective To establish an HPLC method for simultaneously the determination of the isomers of Lornithine-L-aspartate API by UV detector. Methods The chromatography analysis was performed on a Chirex3126（ D）-Penicillamine（ 250 mm × 4. 6 mm,5 μm）. The mobile phase was the mixed solution of1. 5 mmol·L^（-1） copper sulfate and 80 mmol·L^（-1） ammonium acetate with flowrate at 0. 6 m L·min^（-1）. The column temperature is 20 ℃ and the wavelengths were 280 nm. Results The isomers,L-aspartic acid and Lornithine in compound preparations were well separated and the resolution（ R） was more than 1. 5,other impurities do not interfere with the derermination of isomers. D-aspartic Acid and D-ornithine had good linear relationship within the range of 2. 06-19. 92 mg·L^（-1） and 1. 03-19. 69 mg·L^（-1） respectively. The recovery was 96. 9% and 99. 2%. Conclusion The method is simple,rapid,and highly sensitivite,which can be used to control the isomer in L-ornithine-L-aspartate API.