HPLC法同时测定乌腺金丝桃中三种黄酮类化合物的含量

Simultaneous Determination of Three Flavonoids in Hypericum Attenuatum Choisy by HPLC

目的:利用高效液相色谱法同时测定乌腺金丝桃中芦丁、槲皮素、山萘酚的含量。方法:采用Agilent TC-C_(18)(4.6 mm×150 mm,5μm),流动相A为乙腈,B为0.1%磷酸水(v/v);梯度洗脱,流速1.0 mL/min,柱温30℃,检测波长360 nm。结果:在18 min内,乌腺金丝桃中上述3个成分完全分离,芦丁,槲皮素,山萘酚分别在20.0~80.0μg/mL,25.0~80.0μg/mL,14.0~80.0μg/mL的范围内线性关系良好(r值分别为0.999 0,0.999 2,0.999 2);平均加样回收率(n=6)分别为97.88%(RSD=1.16%)、96.57%(RSD=1.89%)、97.68%(RSD=2.56%)。样品中化合物1~3的含量平均值分别为0.02、0.03、0.01 mg/g。结论:该方法简单、快速、结果可靠,可用于同时测定乌腺金丝桃中化合物1~3的含量。

To determinate the contents of rutin,quercetin,and kaempferol in Hypericum attenuatum choisy by RP-HPLC.Methods:The Agilent TC-C18(4.6×150mm,5μm)was used,using gradient elution with a mobile phase of acetonitrile as solvent A and0.1%(v/v)phosphoric acid as solvent B;the flow rate was1.0ml/min;the column temperature was30℃;the determination wavelength was360nm.Results:The three compounds were completely separated within18min.In the range of20.0~80.0μg/ml,25.0~80.0μg/ml,14.0~80.0μg/ml,it was found that there existed a good linear relationship(r=0.9990.0.9992.0.9992).The average recoveries(n=6)were97.88%(RSD=1.16%),96.57%(RSD=1.89%)and97.68%(RSD=2.56%)respectively.The average contents of the three compounds were0.02,0.03,0.01mg/g respectively.Conclusion:The method is simple,fast and reliable,which can be used to determinate the contents of the three compounds in Hypericum attenuatum choisy.