HPLC同时测定乌七祛风散片中4种生物碱的含量

Simultaneous Determination of Four Alkaloids in Wuqi Qufengsan Tablets by HPLC

摘要:

目的: 建立乌七祛风散片(制草乌、三七、断节参、雪上一枝蒿、防风等9味中药组成)中苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱和次乌头碱的HPLC测定方法.方法: 色谱条件,Hanbon-C₁₈色谱柱(4.6 mm×250 mm,5 μm), 流动相:[乙腈-四氢呋喃(25:15)](A)-0.1 mol ·L^-1醋酸铵溶液(每1 000 mL加冰醋酸0.5 mL)(B),流速1.0 mL ·min^-1, 柱温30 ℃,检测波长232 nm.结果: 4种生物碱分离良好,各成分质量浓度与峰面积在测定范围内均呈良好的线性关系(r>0.999 0),加样回收率分别为101.10%(RSD 2.1%),101.09%(RSD 2.2%),102.04%(RSD 1.6%),97.83%(RSD 2.7%),最低检测限分别为3.4,1.7,2.2,0.8 ng.结论: 该HPLC方法稳定可靠、简便易行,可用于同时测定乌七祛风散片中苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱和次乌头碱含量.

Abstract:

Objective:To establish an HPLC method for simultaneous determination of benzoylmesaconine, benzoylaconine, benzoylhypacoitine and hypaconitine in Wuqi Qufengsan tablets. Method: The assay was performed on a Hanbon-C₁₈(4.6 mm×250 mm, 5 μm), mobile phase consisted of acetonitrile-tetrahydrofuran (25: 15, phase A)-0.1 mol · L^-1 ammonium acetate(containing 0.5 mL glacial acetic acid per 1 000 mL, phase B) with gradient elution at the flow rate of 1.0 mL · min^-1, column temperature was 30 ℃, and the detection wavelength was at 235 nm. Result: Excellent chromatographic separation was achieved for four studied alkaloids with good linearity(r>0.999 0), the average recoveries were 101.10%(RSD 2.1%), 101.09%(RSD 2.2%), 102.04%(RSD 1.6%), 97.83%(RSD 2.7%). The limits of detection were 3.4, 1.7, 2.2and 0.8 ng respectively. Conclusion: This method was simple, accurate and reproducible, it can be used to determine the content of benzoylmesaconine, benzoylaconine, benzoylhypacoitine and hypaconitine in Wuqi Qufengsan tablets.