葡萄果实中黄烷- 3 - 醇及其聚合体的HPLC检测

 以‘赤霞珠’葡萄果实(Vitis vinifera L.‘Cabernet Sauvignon’) 为试材, 采用LiChrospher 100 RP-18e色谱柱(250 mm ×410 mm I.D., 5 μm) , 冰乙酸∶水溶液为流动相, 梯度洗脱流速为1.0 mL /min, 柱温30℃, 检测波长280 nm, 成功地分离了( + ) - 儿茶素〔( + ) -catechin, CAT〕、( - ) - 表棓儿茶素〔( - ) -epigallocatechin, EGC〕、( - ) - 表棓儿茶素没食子酸酯〔( - ) -epigallocatechin gallate,EGCG〕、( - ) - 表儿茶素〔( - ) -epicatechin, EC〕和( - ) - 表儿茶素没食子酸酯〔( - ) -epicatechin gallate, ECG〕等5种葡萄果实中最为重要的黄烷- 3 - 醇及其衍生物, 建立了一种快速、准确并可以同时测定葡萄果实中5种黄烷- 3 - 醇及其衍生物的高效液相色谱法; 此外, 经TSK HW-50 (F) 柱分离纯化,间苯三酚存在下酸解后进行HPLC分析, 建立了一种快速、准确测定葡萄果实中多聚黄烷- 3 - 醇平均聚合度、平均分子量及其组成的方法。果实样品检测结果表明: 黄烷- 3 - 醇及其多聚体主要积累于果皮和种子中。

Determination of Monomeric and Polymeric Flavan-3-ols in Grape Berry by High Performance Liquid Chromatography

The grape (Vitis vinifera L. ‘Cabernet Sauvignon’) berry was used as the material. A new method was developed for simultaneously determinating five kinds ofmonomeric flavan-3-ols and their derivatives, including ( + ) -catechin (CAT) , ( - ) -epigallocatechin ( EGC) , ( - ) -epigallocatechin gallate ( EGCG) , ( - ) -epicatechin ( EC) , and ( - )-epicatechin gallate ( ECG) , in the grape berry by high performance liquid chromatography (HPLC). These compounds were efficiently separated directly from the samples through a chromatographic column of 5 μm LiChrospher 100 RP-18e (250 mm ×410mm I.D., Merck)by the mobile phase gradient elution of glacial acetic acid solution at the flow rate of 1.0 mL /min, at the detection wavelength of 280 nm and at 30℃. The polymeric flavan-3-ols had been fractioned and purified on TSK HW-50 ( F) , followed by acid-catalyzed degradation in the presence of phloroglucinol, then the mean degree and the composition of polymeric flavan-3-ols in grape berry were determined by HPLC. The results showed that monomeric and polymeric flavan-3-ols were mainly accumulated in the skins and seeds of grape berry.