茶叶二氢黄酮醇还原酶/无色花青素还原酶反应产物的快速检测方法研究

Method of Determining the Product by DFR/LAR Enzyme Reaction Rapidly in Tea

利用薄层层析方法对儿茶素合成途径中的二氢黄酮醇还原酶/无色花青素还原酶促反应产物儿茶素C进行初步分离,用香草醛显色法进一步定量。以建立该酶促反应产物的快速检测方法。采用薄层层析方法,对薄层层析展开剂配方、及薄层分离产物均进行了研究。该方法检测波长为500nm,展开剂最优配方甲醇:乙酸乙酯:甲酸=8:8:1.5,层析分离所得显色产物为儿茶素C。以儿茶素C为标样,线性范围为50～500μg/ml,加标回收率为91.0%～93.0%之间,RSD为3.39%。建立了薄层层析与香草醛比色法结合检测儿茶素C含量的体系,该体系能够快速测定茶叶中DFR/LAR酶反应产物的含量。

This experiment established the thin layer chromatographic analysis and vanillin colorimetry method to detect catechin,the product of Dihydroflavonol 4-reductase/leucoanthocyanidin reductase enzyme reaction during catechins’ synthesizing,which could determine catechin content rapidly and easily. The thin layer chromatographic developing solvent,and the product separated by the thin layer chromatographic was discussed. As a result,catechin was used as standard samples. Under the condition of 500 nm wavelength,the most superior developing solvent of alcohol： ethyl acetate： formic acid = 8： 8： 1.5,the product which can reveal red is catechin,the linearity range was 50～500μg/ml,and the recovery and the relative standard deviation was in the range of 91.0%～93.0% and 3.39% respectively. the method was developed to determine catechin content rapidly and easily combining the thin layer chromatographic analysis and vanillin colorimetry method.