高效液相色谱法测定黄芪种子中毛蕊异黄酮和异槲皮苷的含量

Determination of Calycosin and Isoquercitrin in Astragalus Seed by HPLC

目的建立高效液相色谱法测定黄芪种子中毛蕊异黄酮和异槲皮苷的含量。方法采用Merck RP-18色谱柱(4.6mm×250mm,5μm),乙腈-0.3%甲酸水为流动相,流速1.0mL·min-1,柱温25℃,检测波长260nm,进样量10μL。结果毛蕊异黄酮和异槲皮苷进样量分别在0.1~0.45μg(r=0.9998)和0.05~0.40μg(r=0.9989)范围内呈良好的线性关系,平均回收率分别为98.45%、98.60%,RSD分别为1.31%、1.45%,黄芪种子中毛蕊异黄酮和异槲皮苷的含量分别为3.7656μg·g-1、2.9843μg·g-1。结论所建立的方法准确度高、重复性好,可用于黄芪种子中毛蕊异黄酮和异槲皮苷的含量测定。

OBJECTIVE To establish a HPLC method for determining the concentration of calycosin and isoquercitrin in astragalus seed.METHODS The chromatographic column was Merck RP-18(4.6mm×250mm,5μm),with acetonitrile-0.3%formic acid water as mobile phase.The flow rate,column tempera ture,detection wavelength and injection volume were 1.0mL·min-1,25℃,260nm,and 10μL,respectively.RESULTS Calycosin and isoquercitrin showed good linearity at 0.1-0.45μg(r=0.9998)and 0.05-0.40μg(r=0.9989)respectively.The average recoveries were 98.45%,98.60%,RSDs value were 1.31%,1.45%,and the contents of calycosin and isoquercitrin were 3.7656μg·g-1,2.9843μg·g-1,respectively.CONCLUSION The HPLC method is accurate and reproducible,and suitable for the content determination of calycosin and isoquercitrin in astragalus seed.