HPLC与紫外-可见分光光度法对翻白草中黄酮类成分含量测定

Determination of flavonoid content in Potenlilla discolor by HPLC and UV-VIS spectrophotometry

目的 建立测定翻白草中黄酮类成分含量的最佳方法。方法 先建立高效液相色谱法(HPLC)测定翻白草中的4种黄酮类物质含量的方法,其中流动相为乙腈与0.1%甲酸溶液,并采用梯度洗脱,柱温设为30℃,色谱柱为InertSustain C18(150 mm×4.6 mm, 5μm),检测波长设为360 nm,并对不同批次的翻白草中的4种黄酮类物质含量进行检测;选取芦丁作为对照品,以UV-VIS法测定翻白草中总黄酮含量。结果 不同产地的翻白草含有的黄酮类成分含量差异较大,总黄酮含量的差异也较大,建立的HPLC法与紫外-可见分光光变法(UV-VIS)法具有很好地重复性,样品稳定性好,且仪器精密度高,两种检测方法相结合能很好的把控翻白草品质。结论 以UV-VIS法对翻白草中的总黄酮进行测定,并采用HPLC法测定翻白草中含有的槲皮苷、芦丁、山柰酚和槲皮素,方法较为简便,且有很好的重复性、稳定性,精密度高,能很好地控制翻白草的品质,并为临床研究及其药理活性研究提供依据。

Objective To establishthe best method of determining the total flavonoid content of Potenlilla discolor.Methods HPLC method was established to determine the content of 4 kinds of flavonoids in Potenlilla discolor.The mobile phase was acetonitrile and 0.1%formic acid solution with gradient elution,the column temperature was set at 30℃,and the detection wavelength was set at 360 nm.The content of flavonoids in different batches of Potenlilla discolor was determined by UV-VIS method,and rutin was selected as the control.Results The contents of flavonoids and total flavonoids of Potenlilla discolor in different producing areas were different.The established HPLC method and UV-VIS method have good reproducibility,with good sample stability and high instrument precision,and the combination of the two detection methods could control the quality of Flavonoids in Potenlilla discolor very well.Conclusion The UV-VIS method for the determination of total flavonoids and HPLC method for the determination of quercetin,rutin,kaempferol and quercetin in Potenlilla discolor are relatively simple,with good reproducibility,stability and precision,and can control the quality of Potenlilla discolor well and provide the basis for the clinical study and its pharmacological activity research.