HPLC同时测定二黄汤≤1000Da提取液中6种成分的含量

Simultaneous Determination of Six Components in the ≤1000 Da Extract of Erhuang Decoction by HPLC

目的：优化HPLC条件,建立同时测定二黄汤≤1 000 Da提取液中6种有效成分（黄芩苷、黄芩素、巴马汀、小檗碱、甘草苷、甘草酸）含量的方法。方法：采用RP-HPLC,Diamonsil C18色谱柱（4.6 mm×250 mm,5μm）,流动相乙腈-乙酸铵溶液（0.02 mol·L^-1,冰乙酸调pH 4.0）梯度洗脱,流速0.5 mL·min^-1,检测波长280,265,250 nm,柱温25℃,进样量20μL。结果：黄芩苷、黄芩素、巴马汀、小檗碱、甘草苷、甘草酸在各自的线性范围内与色谱峰面积呈良好的线性关系,回收率在95%~105%,精密度、稳定性的RSD均〈2.0%。结论：该方法可快速简便的测定二黄汤中6种有效成分的含量,可为控制二黄汤方药制剂质量标准评价提供依据。

Objective： The study was performed optimize the HPLC conditions and to establish a method to determine content of six effective components in the ≤ 1 000 Da extract of Erhuang decoction（baicalin,baicalein,palmatine,berberine,liquiritin,glycyrrhizic acid） simultaneously. Method： The RP-HPLC method used for the determination,on a chromatographic column of Diamonsil,C18column（4. 6 mm × 250 mm,5 μm）;with the was used mobile phase composed of acetonitrile： ammonium acetate solution（0. 02 mol·L^-1,acetic acid pH 4. 0）,in gradient elution; the flow rate was fixed at 0. 5 mL·min^-1; the detection wavelength was set at 280,265,250 nm; the column temperature： was maintained at 25 ℃; the injection volume was 20 μL. Result： There is a good linear relationship between the chromatographic peak area and concentration of baicalin,baicalein,palmatine,berberine,liquiritin,glycyrrhizic acid in the linear range,and the recovery are all in the range of 95%-105%,with all the RSD of the recovery rate less than 2%. Conclusion： The method is rapid and simple for the content determination of six components content determination in the ≤1 000 Da extract of Erhuang decoction.