高效液相色谱法同时测定仙鹤草中6种黄酮

Determination of Six Flavonoids in Agrimonia pilosa Ledeb by HPLC

目的：建立超声辅助萃取-高效液相色谱法分离仙鹤草中芦丁、花旗松素、香豆素、木犀草素、山奈酚和芹菜素6种黄酮及其含量测定的方法。方法：采用Phenomenex？C18色谱柱（150？mm×4.6？mm,5？μm）分离6种成分;流动相为甲醇-0.1%醋酸溶液,梯度洗脱;流速0.8？mL/min,紫外检测波长320？nm,柱温35℃。结果：6种成分在15？min内均达到基线分离,线性关系良好（r不低于0.999 1,n=6）,平均回收率均在97.78%～104.4%之间（相对标准偏差小于1.20%,n=3）。结论：本实验采用超声波法提取,方法简单、快速、经济、可靠,可用于仙鹤草的质量监控。

Objective： To establish a high performance liquid chromatography（HPLC）method for separation and quantification of rutin, taxifolin, coumarin, luteolin, kaempferol and apigenin in Agrimonia pilosa Ledeb by ultrasonic-assisted extraction. Methods： The separation of six flavonoids was performed on Phenomenex C18column（150mm×4.6mm,5μm） using step gradient elution. The mobile phase was composed of methanol and 0.1% acetic acid at a flow rate of 0.8 mL/min. The UV detection wavelength was 320nm and the column temperature was set at 35℃. Results：Good separation of rutin, taxifolin, coumarin, luteolin, apigenin and kaempferol was achieved within 15min.Calibration curves of all the six components showed an excellent linear relationship（r≥0.9991,n =6）.The average recoveries were between 97.78%and 104.4% with a relative standard deviation（RSD）of〈1.20%.Conclusion：This method, based on ultrasonic-assisted extraction, is simple,fast,economical and reliable.