HPLC法同时测定杜仲叶中4种成分的含量

摘要

目的:建立高效液相色谱法同时测定杜仲叶中4种成分含量的方法.方法:采用C18色谱柱(200mm×4.6 mm,10 μm),以甲醇-0.1％磷酸溶液为流动相,梯度洗脱(甲醇:0～2 min,22％;2～6 min,22～60％;6～20 min,60％;20～ 24 min,60 ～ 22％;24～28 min,22％);流速0.8 mL/min;柱温30℃;检测波长绿原酸为327 nm,芦丁、槲皮素、山奈酚为360 nm.结果:绿原酸、芦丁、槲皮素、山奈酚进样量分别在16.4～98.4 μg/mL,15.6～93.6 μg/mL,15.18 ～ 91.07 μg/mL,15.7～94.2μg/mL范围内线性关系良好,平均加样回收率分别为101.0％、98.6％、102.9％、103.3％.结论:建立的方法符合方法学验证要求,可用于杜仲叶中绿原酸、芦丁、槲皮素、山奈酚4种成分的含量测定.%Objective:To establish an HPLC method for simultaneous determination of the contents of 4 compounds in the leaves of Oliv..Methods:C1s column (200 mm x 4.6 mm,10 μm) was adopted.The mobile phase was methanol-1％ phosphoric acid solution with gradient elution (methanol:0-2 min,22％;2-6 min,22-60％;6-20 min,60％;20-24 min,60-22％;24-28 min,22％)at a flow rate of 0.8 mL/min and the temperature of column was kept at 30℃.Results:Chlorogenic acid,rutin,quercetin and kaempferol had a good linear relationship in the range of 16.4 ～ 98.4,15.6 ～ 93.6,15.18-91.07 and 15.7-94.2 μg/mL,and the average recoveries of 4 compounds were 101.0％,98.6％,102.9％ and 103.3％,respectively.Conclusion:This method conforms to the validation requirement of methodology,and it can be used for the content determination of chlorogenic acid,rutin,quercetin and kaempferol in the Eucommia ulmoides leaves.