应用高速逆流色谱结合Sephadex LH-20和半制备HPLC对茶色素中三种主要茶黄素(茶黄素,TF1;茶黄素单没食子酸酯,TF2+TF3;茶黄素双没食子酸酯,TF4)进行了分离纯化,并以它们为标样,建立了茶色素中三种主要茶黄素的HPLC定量法。精密度和加样回收率实验结果表明,TF1、TF2+TF3和TF4的变异系数分别为2.6’8.5%、2.0’3.8%和2.0’3.9%,三者总量的变异系数为2.2’4.1%。应用该法对Sigma公司的混合茶黄素标样测定表明,其相对误差为3.4%。对自制茶色素中茶黄素类含量分析表明,茶黄素类总量可达20.5%、其中TF1、TF2+TF3和TF4的含量则分别为6.0%、9.1%和5.4%。茶色素中茶黄素类HPLC定量方法的建立,对于茶色素质量控制具有重要的实际价值,并有助于茶色素的开发以及药理功能的深入研究。
In this study, theaflavin(TF1), theaflavin monogallate(TF2+TF3) and theaflavin digallate(TF4) in tea pigments are isolated and purified with high speed counter current chromatography combined with Sephadex LH-20 and semi-preparative HPLC. With the purified theaflavin, the standard curves have been set up by HPLC for theaflavins quantitation. The results on the accuracy and recovery determination showed that the coefficient of variation of TF1, TF2+TF3, TF4 and total theaflavins range from 2.6’8.5%, 2.0’3.8%, 2.0’3.9% and 2.2’4.1% respectively. Using HPLC quantitation, the relative error of total theaflavins amounts in the standard theaflavins from Sigma company is 3.4%. The total amounts of theaflavins in tea pigments are 20.5%, in which TF1, TF2+TF3 and TF4 are 6.0%, 9.1% and 5.4% respectively. This method is useful to quantify theaflavins and to control the quality of tea pigments, which could promote the studies on their pharmacological effects.
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