HPLC-ELSD法同时测定参芪颗粒中6个成分的含量

目的:建立HPLC-ELSD法同时测定参芪颗粒中党参炔苷、芒柄花素、黄芪皂苷Ⅰ、黄芪皂苷Ⅱ、黄芪皂苷Ⅲ和黄芪甲苷(黄芪皂苷Ⅳ)的含量测定方法。方法:采用Agilent C₁₈色谱柱(4.6 mm×250 mm,5μm),以乙腈-0.5%(v/v)醋酸水溶液为流动相,进行梯度洗脱,流速1.0 mL·min^-1,柱温35℃;使用蒸发光散射检测器,载气气压为2.76×10⁵ Pa,漂移管温度为70℃。结果:在34 min内,参芪颗粒中6个成分(党参炔苷、芒柄花素、黄芪皂苷Ⅰ、黄芪皂苷Ⅱ、黄芪皂苷Ⅲ和黄芪甲苷)完全分离,峰面积的对数与进样量的对数呈良好的线性关系(r=0.9995~0.9999);平均回收率分别为96.7%、99.5%、99.5%、98.7%、99.6%、97.4%。3批样品中党参炔苷、芒柄花素、黄芪皂苷Ⅰ、黄芪皂苷Ⅱ、黄芪皂苷Ⅲ和黄芪甲苷含量分别在0.10~0.13、0.0086~0.0121、0.014~0.018、0.056~0.060、0.0023~0.0041、0.055~0.066 mg·g^-1。结论:该方法经方法学验证可用于参芪颗粒的质量控制。

Objective:To establish an HPLC-ELSD method for simultaneous determination of lobetyolin, formononetin and astragalosidesⅠ,Ⅱ,Ⅲ,Ⅳ in Shenqi granules. Methods:The separation was performed on an Agilent C₁₈ column(250 mm×4.6 mm, 5μm)with gradient elution at a temperature of 35℃. The acetonitrile and 0.5%(v/v)acetic acid was used as the mobile phase with a flow rate of 1.0 mL·min^-1. The detector was ELSD, of which the drift tube temperature was maintained at 70℃ and the pressure of nebulizing gas was 2.76×10⁵ Pa. Results:The six compounds(lobetyolin,formononetin and astragalosides Ⅰ,Ⅱ,Ⅲ,Ⅳ)were completely separated within 34 min. The regression equation showed good liner relationship between logarithm of the peak area and logarithm of the content of each compound(r=0.9995~0.9999). The average recoveries were 96.7%, 99.5%, 99.5%, 98.7%, 99.6%, and 97.4%, respectively. The contents of lobetyolin, formononetin and astragalosides Ⅰ, Ⅱ, Ⅲ, Ⅳ in 3 samples were 0.10-0.13, 0.0086-0.0121, 0.014-0.018, 0.056-0.060, 0.0023-0.0041 and 0.055-0.066 mg·g^-1, respectively. Conclusion:The established method is validated by methodology, and is suitable for the quality control of Shenqi granules.