HPLC测定清肝活血方中4种黄酮的含量

 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Agilent Zorbax Eclipse XDB-C₁₈柱(4.6 mm×250 mm, 5 μm)，流动相为甲醇-0.5%甲酸水梯度洗脱，流速为1.0 mL·min-1，检测波长为276 nm。结果 葛根素的线性范围为0.12~1.98 g·L-1(r=0.999 8)，平均回收率为99.87%，RSD=2.04%；黄芩苷的线性范围为0.24~3.82 g·L-1(r=0.999 9)，平均回收率为99.74%，RSD=1.84%；黄芩素的线性范围为0.01~0.14 g·L-1(r=0.999 6)，平均回收率为99.64%，RSD=1.96%；汉黄芩素的线性范围为0.007~0.12 g·L-1(r=0.999 5)，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。

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

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×250 mm, 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 baicalein. The calibration curve was linear (r=0.999 6) in the range of 0.01-0.14 g·L-1and the average recovery was 99.64% (RSD=1.96%) for baicalin. The calibration curve was linear (r=0.999 5) in the range of 0.007-0.12 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.