固相萃取-高效液相色谱法测定清肝降压胶囊中葛根素的含量

Determination of Puerarin in Qinggan Jiangya Capsule by Solid Phase Extraction-High Performance Liquid Chromatography

目的以固相萃取小柱净化除杂,建立高效液相色谱法测定清肝降压胶囊中葛根素含量。方法样品经HyperSep C18固相萃取小柱处理,采用Diamonsil C18(250 mm×4.6 mm,5μm)色谱柱分离,以甲醇-0.01%KH2PO4(28∶72)为流动相,检测波长为250 nm,柱温40℃,体积流量1.0 mL/min。结果清肝降压胶囊中葛根素含量测定的标准曲线方程为Y=18.1362X-17.6029(r=0.9998),在1.68~83.84 mg/L范围内葛根素线性关系良好,检出限为0.17 mg/L,3个不同剂量的平均加样回收率为97.05%,RSD为0.43%。结论本法的重复性和精密度良好,符合分析方法要求。通过预处理去除了部分杂质,测定葛根素快速准确,专属性强,重现性好,适用于清肝降压胶囊的质量控制。

Objective To establish the determination method of puerarin in Qinggan Jiangya capsule by solid phase extraction and high performance liquid chromatography.Methods Samples were pretreated by HyperSep C18 solid phase column to remove water soluble substances and analyzed by high performance liquid chromatography with Diamonsil C18(250mm×4.6mm,5μm)column and a mixture of methanol-0.01%KH2PO4(28:72)solution as the mobile phase.The detection wavelength was 250 nm,flow rate was 1.0 mL/min and column temperature was 40℃.Results Puerarin was well isolated.The determination of puerarin in Qinggan Jiangya capsule had the regression equation as Y=18.1362X-17.6029(r=0.9998).The calibration graph of Puerarin was linear within 1.68~83.84 mg/L.The detection limit of Puerarin was 0.17mg/L and the average recovery rate of low,medium and high doses was 97.05%(RSD was 0.43%).Conclusion This method show good accuracy and repeatability,and met the requirement of analytical method.The method can effectively eliminate the negative interference with good specificity,sensitivity,accuracy and suitable for the quality control of puerarin in qingganjiangya capsule.