葡萄果实中黄烷-3-醇及其聚合体的HPLC检测

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

以‘赤霞珠’葡萄果实(VitisviniferaL.‘CabernetSauvignon’)为试材,采用LiChrospher100RP-18e色谱柱(250mm×4·0mmI·D·,5μm),冰乙酸∶水溶液为流动相,梯度洗脱流速为1·0mL/min,柱温30℃,检测波长280nm,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(-)-epigallocatechingallate,EGCG、(-)-表儿茶素(-)-epicatechin,EC和(-)-表儿茶素没食子酸酯(-)-epicatechingallate,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TSKHW-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。

The grape （ Vitis vinifera L. ‘ Cabernet Sauvignon＇ ） berry was used as the material. A new method was developed for simultaneously determinating five kinds of monomeric 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 sam- ples through a chromatographic column of 5 μm LiChrospher 100 RP-18e （250 mm ×4. 0mm 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.