HPLC测定清肝活血方中4种黄酮的含量

Determination of Puerarin,Baicalin,Baicalein and Wogonin in Qinggan Huoxue Recipe by HPLC

目的建立HPLC同时测定清肝活血方中葛根素、黄芩苷、黄芩素和汉黄芩素含量的方法。方法采用HPLC进行分析，色谱柱为AgilentZorbaxEclipseXDB—C18柱（4．6mm×250mm，5μm），流动相为甲醇-0．5％甲酸水梯度洗脱，流速为1．0mL·min^-1，检测波长为276nm。结果葛根素的线性范围为0．12～1．98g·L^-1（r=0．9998），平均回收率为99．87％，RSD=2．04％；黄芩苷的线性范围为0．24～3．82g·L^-1（r=0．9999），平均回收率为99．74％，RsD=1J84％；黄芩素的线性范围为n01～n14g·L^-1（r=n9996），平均回收率为99．64％，RSD=1．96％；汉黄芩素的线性范围为0.007～0．12g·L^-1（r=0．9995），平均回收率为100．83％，RSD=2．37％。结论本方法操作简便，测定结果准确可靠，可用于清肝活血方的质量控制。

OBJECTIVE To establish a quantitative method simultaneously for the determination of puerarin, baicalin, baicalein and wogonin in Qinggan Huoxue recipe by HPLC. METHODS The separation was performed on an Agilent Zorbax Eclipse XDB-C18 column （4. 6 mm × 250mm, 5 μm）. The mobile phase consisted of methanol and 0. 5% methanoic acid with gradient elution, at the flow rate of 1.0 mL · min ^-1 The wavelength was at 276 nm. RESULTS The calibration curve was linear （ r = 0. 999 8） in the range of 0. 12 - 1.98 g ·L-1 and the average recovery was 99. 87% （ RSD = 2. 04% ） for puerourin. The calibration curve was linear （r =0. 999 9） in the range of 0. 24 -3. 82 g · L-1 and the average recovery was 99. 74% （RSD = 1.84% ）, for baiealein. The calibration curve was linear （r = 0. 999 6） in the range of 0. 01 - 0. 14 g · L^- 1 and the average recovery was 99. 64% （ RSD = 1.96% ） for baiealin. Thecalibration curve was linear （ r = 0. 999 5） in the range of 0. 37 - 0. 12 g · L^- 1 and the average recovery was 100. 83% （ RSD = 2. 37% ） for wogonin. CONCLUSION The method was sensitive, accurate and can be used for the quality control of Qinggan Huoxue recipe.