软枣猕猴桃多糖的分离与纯化及其分子质量的测定

Isolation, Purification and Molecular Weight Determination of Polysaccharides from Actinidia arguta Sieb. et Zucc. Fruits

对通过水提醇沉法提取、Sevag法脱蛋白得到的软枣猕猴桃多糖进行分离纯化,利用DEAE-纤维素52离子交换层析对软枣猕猴桃多糖进行初步分离,得到4种软枣猕猴桃多糖组分;利用葡聚糖凝胶对软枣猕猴桃多糖组分进一步分离的最佳条件为葡聚糖凝胶型号为Sephadex G-200、层析柱规格为D1.1cm×100cm、多糖质量浓度为10g/L。以此条件分离,得到含量较高的软枣猕猴桃多糖-Ⅱ纯品。通过Sephadex G-200凝胶柱层析鉴定软枣猕猴桃多糖-Ⅱ是均一的纯多糖,通过紫外光谱鉴定软枣猕猴桃多糖-Ⅱ不含核酸及蛋白。同时测得软枣猕猴桃多糖-Ⅱ的分子质量为83733D。

Polysaccharides from Actinidia arguta Sieb.et Zucc.were extracted by hot-water extraction,ethanol precipitation,protein removal by the Sevag method and then purified by DEAE-cellulose-52 ion exchange column chromatography.Four polysaccharide fractions were obtained.The best purification of Fraction Ⅱ was achieved by chromatographic separation on a Sephadex G-200 column（D1.1 cm × 100 cm） at a sample concentration of 10 g/L.As a result,a homogenous pure polysaccharide was obtained.The results of UV spectroscopic analysis revealed that the purified polysaccharide contained neither proteins nor nucleic acids.Moreover,the molecular weight of the purified polysaccharide was measured to be 83733 D.