UPLC法测定不同产地秋海棠中5种药效成分的含量

Simultaneous Determination of 5 Active Ingredients in Begonia Grandis Dry from Different Habitats by UPLC

目的:建立秋海棠药材中原儿茶酸、原儿茶醛、儿茶素、表儿茶素、芦丁的UPLC同时检测方法,测定和比较了不同产地秋海棠中5种药效成分的含量。方法:用70%甲醇回流提取样品中药效成分,采用Waters ACQUITY HSS T3(2.1 mm×100mm,1.8μm)色谱柱,0~12 min乙腈-0.2%冰醋酸水溶液(5→25∶95→75),12~30 min乙腈-0.2%冰醋酸水溶液(25→50∶75→45),流速:0.2 ml·min^(-1),检测波长:280 nm,柱温:30℃,进样体积:1μl,应用Waters ACQUITY UPLC H-CLASS超高液相色谱仪进行测定。结果:秋海棠药材中5种药效成分的分离效果良好,在相应的浓度范围内与峰面积有良好线性关系,相关系数大于0.99,平均回收率分别为96.92%~99.75%(n=6)。结论:该方法应用于研究快速、准确、重复性好,可为秋海棠的质量控制提供快速准确的检测方法。

Objective： To establish a method for the determination of protocatechuic acid,protocatechuic,catechin,epicatechin and rutin in Begonia grandis Dry. Methods： The 5 active ingredients in the samples were extracted by 70% methanol and analyzed by ultra performance liquid chromatography（ UPLC）. Chromatographic separation was performed on a Waters ACQUITY HSS T3（ 2.1 mm× 100 mm,1.8 μm） column with gradient elution,and the mobile phase was solvent A： acetonitrile and solvent B： 0.2% glacial acetic acid（ 0-12 min： A： 5→25 and B： 95→75; 12-30 min： A： 25→50 and B： 75→45）. The effluent was monitored by a VWD detector set at 280 nm. The flow rate was set at 0.2ml / min and the injection volume was 1 μl. The column temperature was maintained at 30℃.Results： The calibration curves were all linear（ r≥0.99） over the test ranges. The average recovery was 96.92%-99.75%（ n = 6）.Conclusion： The established method can rapidly obtain an accurate and reproducible result,which can be used to control the quality of Begonia grandis Dry.