HPLC-DAD-ELSD法测定补阳还五汤中8个活性成分含量

Determination of eight active components in Buyang Huanwu Decoction by HPLC-DAD-ELSD

目的建立高效液相色谱-二极管阵列检测器-蒸发光散射检测器(HPLC-DAD-ELSD)法同时测定补阳还五汤中羟基红花黄色素A、芍药苷、毛蕊异黄酮苷、阿魏酸、芒柄花苷、毛蕊异黄酮、芒柄花黄素和黄芪甲苷8个活性成分含量的方法。方法采用Agilent Eclipse XDB-C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.1%甲酸为流动相,梯度洗脱,流速1.0 ml/min,柱温30℃,检测波长230 nm(芍药苷)、254 nm(毛蕊异黄酮苷、芒柄花苷、毛蕊异黄酮、芒柄花黄素)、322 nm(阿魏酸)和403 nm(羟基红花黄色素A),蒸发光散射检测器漂移管温度60℃,载气流速1.6 L/min。结果羟基红花黄色素A、芍药苷、毛蕊异黄酮苷、阿魏酸、芒柄花苷、毛蕊异黄酮、芒柄花黄素和黄芪甲苷分离度良好,线性关系符合要求(r=0.9940~0.9999),平均回收率为97.8%~101.4%,RSD为1.28%~3.70%。结论该方法简便、稳定、重复性良好,可用于补阳还五汤的质量控制。

Objective To establish an HPLC-DAD-ELSD method for the simultaneous determination of eight main active components in Buyang Huanwu Decoction,including hydroxysafflor yellow A,paeoniflorin,calycosin glycoside,ferulic acid,ononin,calycosin,fermononetin and astragaloside.Methods Agilent Eclipse XDB-C18 column(250 mm×4.6 mm,5μm)was used with acetonitrile-0.1%formic acid as the mobile phase.The flow rate was 1.0 ml/min;the column temperature was 30℃;the detection wavelengths were 230 nm(paeoniflorin),254 nm(calycosin glycoside,ononin,calycosin,fermononetin),322 nm(ferulic acid)and 403 nm(hydroxysafflor yellow A);the drift tube temperature of the evaporative light scattering detector was 60℃;the carrier gas flow rate was 1.6 L/min.Results Under these conditions,the separation of hydroxysafflor yellow A,paeoniflorin,calycosin glycoside,ferulic acid,ononin,calycosin,fermononetin and astragaloside was good,and the linear relationship was in line with the requirements(r=0.9940-0.9999).The average recovery was 97.8%-101.4%(RSD was 1.28%-3.70%).Conclusion The method is simple,stable and reproducible,and can be used for the quality control of Buyang Huanwu Decoction.