应用高效液相色谱法测定茶叶多糖.pdfVIP

2006, Vol. 27, No. 04 177 聂少平，谢明勇 * ，申明月，王远兴 (南昌大学 食品科学教育部重点实验室，江西 南昌 330047) ：使用Sephadex G-100凝胶柱纯化制备得到茶叶多糖，用高效凝胶渗透色谱法 (HPGPC)鉴定了茶叶多糖的 纯度并测定其分子量；同时，使用不同浓度的NaCl溶液作为流动相并用已知分子量的右旋葡聚糖作为标准曲线研 TM 究了茶叶多糖在凝胶色谱柱(Ultrahydrogel 500)上的色谱行为，研究发现，由于茶叶多糖的分子量随着流动相的 离子浓度的改变而改变，高效凝胶渗透色谱法是不适合用作测定茶叶多糖的分子量的方法的。然后，本研究以自 制茶叶多糖纯品作为标准品，以UltrahydrogelTM 500(7.8×300mm)为固定相，0.6ml/min蒸馏水为流动相，折光示 差检测，对多种茶叶中茶叶多糖含量进行了测定，回收率在80.4%～93.2% (RSD=5.42%)之间，回归方程为： -6 C=0.2503×10 A-0.0087，r=0.9997，其浓度在0.604～6.04mg/ml范围内呈良好的线性关系。结果显示，该方法 是简单、准确、可靠的，适合于作为茶叶多糖原材料和产品质量控制的方法。 ：高效凝胶渗透色谱法；茶叶多糖；含量测定 Determination of Tea Polysaccharide by High Performance Liquid Chromatography NIE Shao-ping，XIE Ming-yong*，SHEN Ming-yue，WANG Yuan-xing (Key Laboratory of Food Science of Ministry of Education, Nanchang University, Nanchang 330047, China) Abstract：Tea polysaccharide was purified by Sephadex G-100 gel filtration, and its purity and molecular weight (Mw) were determined by high performance gel permeation chromatography (HPGPC). The GPC elution behavior of tea polysaccharide on UltrahydrogelTM 500(7.8×300mm)column was studied by using mobile phase with various concentrations of NaCl. Dextran T- system was found to be unsuitable as Mw calibration standards in GPC because the Mw of tea polysaccharide was changed with the change of ionic concentration in the mobile phase. Thus, the purified tea polysaccharide obtained in this study was standardized to be used instead as the calibration standards for the determination and comparison of tea polysaccharides in tea samples collected from different geographic locations and species varieties. The GPC was run at 35℃on