膜过滤绿茶多糖的系统分级纯化及免疫活性分析

Systematic Separation, Purification of Water-soluble Green Tea Polysaccharides and Their Immunological Activities

采用150kD、20kD、6kD的超滤膜和对水溶性茶多糖进行分级和浓缩,得到的三部分截留液分别经过DEAE-52纤维素离子交换层析和丙烯葡聚糖凝胶Sephacryl S-300柱层析系统分离、纯化,共得到了20多个分级组分;HPGPC-ELSD鉴定各多糖组分的纯度及分子量分布,结果表明得到5个均一多糖组分;用小鼠巨噬细胞系Raw264.7检测免疫活性,发现对小鼠巨噬细胞Raw264.7有显著活性的多糖组分大多集中在20kD左右,大多为不均一多糖。

Water-soluble green tea polysaccharides were separated and purified by Ultrafiltration membranes with pore diameter of 150 kD, 20 kD, 6 kD in turn. All the three fractions were further separated and purified by DEAE-cellulose ion-exchange chromatography and Sephacryl S-300 column chromatography, and more than 20 green tea polysaccharide components totally were obtained. Their purity and molecular weight distribution were determined with high performance gel permeation chromatography （HPOPC） equipped with ELSD detector, and five components of homogeneous polysaccharide were obtained. The immunological activities of these green tea polysaccharides were determined by stimulated phagocytosis and nitric oxide production on RAW264.7 macrophage cell line of mice. Results showed that the tea Polysaccharides with 20 kD MW possessed significant immunological activities on NO production or phagocytosis of mice macrophage Raw 264.7 cells.