高效液相色谱法测定测定门冬氨酸鸟氨酸原料中S-3-氨基哌啶-2-酮、精氨酸和谷氨酸的含量

Determination of S-3-2-ketone amino piperidine、arginine and fumaric acid in ornithine aspartate by high performance liquid chromatography

目的采用高效液相色谱法建立同时测定门冬氨酸鸟氨酸中S-3-氨基哌啶-2-酮(杂质A)、精氨酸和谷氨酸含量的方法。方法色谱柱Sepax HP-Amino(4.6×250 mm,5μm),流动相为0.05mol.L-1磷酸二氢钾-乙腈(42∶58)混合后用磷酸调pH至5.2,检测波长为200nm。结果杂质A浓度在0.1950～97.49μg.mL-1范围内与峰面积呈良好的线性关系,r为1.000;精氨酸浓度在1.006～20.12μg.mL-1范围内与峰面积呈良好的线性关系,r为1.000;富马酸浓度在0.04258～21.29μg.mL-1范围内与峰面积呈良好的线性关系,r为1.000;杂质A加样回收率的平均值为99.8%,RSD=1.9%;精氨酸加样回收率的平均值为100.4%,RSD=1.7%;富马酸加样回收率的平均值为100.9%,RSD=1.5%。结论本方法简便、准确可靠,适用于门冬氨酸鸟氨酸中杂质A、精氨酸和富马酸的控制。

OBJECTIVE To establish an HPLC method for determining the contents of S-3-2-ketone amino piperidine、arginine and fumaric acid in ornithine aspartate.METHODS The condition of detection were as following： Sepax HP-Amino（250mm × 4.6mm,5μm）,the mobile phase consisted of 0.05mol.L-1 Monobasic Potassium Phosphate and acetonitrile（42∶ 58）（Adjusted pH to 5.2 with phosphoric acid after mixing）,the UV detection wavelength was 200nm.RESULTS The calibration curves were linear in the range of 0.1950 ～ 97.49μg·mL^-1 for impurity A（r = 1.000）;the calibration curves were linear in the range of 1.006 ～ 20.12μg·mL^-1 for arginine（r = 1.000）;the calibration curves were linear in the range of 0.04258 ～ 21.29μg·mL^-1 for fumaric acid（r = 1.000）.The average recoveries of impurity A were 99.8%（RSD = 1.9%）;the average recoveries of arginine were 100.4%（RSD = 1.7%）;the average recoveries of fumaric acid were 100.9%（RSD = 1.5%）.CONCLUSION The method is simple,accurate,reliable and can be used for the quality control of impurity A、arginine and fumaric acid in ornithine aspartate.