HPLC-DAD法测定降脂宁颗粒中二苯乙烯苷、大黄素和大黄素甲醚的含量

The Assay of Stilbene Glycoside,Emodin and Physcion in Jiangzhining Granules by Using HPLC-DAD Method

本研究旨在建立降脂宁颗粒中制何首乌成分2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、大黄素和大黄素甲醚含量测定方法。采用HPLC-DAD法,以Caprisil C18-B(4.6mm×250mm,5μm)色谱柱;测定二苯乙烯苷的流动相为乙腈-水(20:80),流速为1.0 mL/min;检测波长为320 nm;柱温30℃;测定大黄素和大黄素甲醚的流动相为乙腈-0.1%磷酸溶液,梯度洗脱,流速为1.0 mL/min,检测波长222 nm。结果表明,2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、大黄素和大黄素甲醚分别在0.042~0.422μg、0.008~0.082μg和0.011~0.109μg的质量范围内与峰面积线性关系良好,相关系数均为0.9999。本研究建立的方法精密度、稳定性(24 h)、重复性的RSD均小于2.0%(n=6),平均回收率依次为99.2%、95.5%和94.8%。3批样品中制何首乌成分含量差异较大。本方法简单易操作,准确高效,可为降脂宁颗粒中制何首乌成分含量测定提供参考。

This study aims to establish a HPLC-DAD method for the determination of 2,3,5,4'-tetrahydroxystilbene-2-O--D-glucoside,emodin and physcion in Jiangzhining granules.The separation was performed on a Caprisil C18-B(4.6 mm×250 mm,5μm)column by HPLC.The mobile phase for the determination of stilbene glycoside was acetonitrile-water(20:80),and the flow rate was 1.0 mL/min.The detection wavelength was 320 nm.Column temperature 30℃.The mobile phase for the determination of emodin and physcion was acetonitrile-0.1%phosphoric acid solution with gradient elution.The flowrate was 1.0 mL/min and the detection wavelength was 222 nm.The mass of 2,3,5,4'-tetrahydroxystilbene-2-O--D-glucoside,emodin and physcion showed good linearity with the peak area in the range of 0.042~0.422 g,0.008~0.082 g and 0.011~0.109 g,respectively.And the correlation coefficient was 0.9999.The RSDs of precision,stability(24 h)and repeatability experiments were all less than 2.0%(n=6).The recoveries were 99.2%,95.5%and 94.8%,respectively.The contents of the components in three batches of samples were quite different.The method is simple,easy to operate,accurate and efficient,and can provide a reference for the determination of the content of prepared Polygoni Multiflori Radix in Jiangzhining Granules.