HPLC-ELSD法测定花丹安神合剂中N-甲基-反式-4-羟基-L-脯氨酸的含量

Determination of the content of N-methyl-trans-4-hydroxy-L-proline in Huadan’anshen Mixture by HPLC-ELSD

文章建立花丹安神合剂中N-甲基-反式-4-羟基-L-脯氨酸的含量测定方法。方法:应用高效液相色谱法测定,色谱柱为YMC-Pack Polyamine II色谱柱(4.6 mm×250 mm,5μm);流动相为75%乙腈;流速为1.0 mL/min;柱温为30℃;检测器为ELSD检测器(漂移管温度48℃,气体流量1.8 L/min,增益1)。结果:N-甲基-反式-4-羟基-L-脯氨酸在5~20μg范围内峰面积与浓度呈良好的线性;加样回收率为96.76%,RSD为1.53%。结论:该方法简便易行、准确、重复性好,可用于花丹安神合剂的含量测定及质量控制。

This paper establishes the determination method of the content of N-methyl-trans-4-hydroxy-L-proline in Huadan’anshen Mixture. Method: High performance liquid chromatography(HPLC)method is applied, with YMCPack Polyamine II column(4.6 mm×250 mm, 5 μm), 75% acetonitrile as the mobile phase, flow rate being 1.0 mL/min,column temperature being 30 ℃, and detector being the ELSD(the drift tube temperature being 48 ℃, the gas flow rate being 1.8 L/min and the gain being 1). Results: The peak area of N-methyl-trans-4-hydroxy-L-proline is in good linearity with concentration in the ranger of 5-20 μg;The sample recovery rate is 96.76%;RSD is 1.53%. Conclusion:The method is simple, accurate and reproducible, and can be used for the determination of the content and quality control of Huadan’anshen Mixture.