红花多糖的分离纯化及单糖组成分析

Isolation,purification and monosaccharide composition analysis of polysaccharides from safflower

目的:从红花中分离纯化得到红花多糖(SPS),并对其进行相对分子质量测定及单糖组成分析。方法:红花经热水提取、乙醇沉淀、冷冻干燥、Sevag法除蛋白与大孔吸附树脂HPD-100脱色,再经DEAE-52纤维素柱和Sephadex G-100凝胶色谱分离纯化得到红花多糖。运用紫外光谱鉴定纯度,高效凝胶过滤色谱法(HPGPC)分析其相对分子质量,并借助衍生化气相色谱法(GC)和红外光谱(FT-IR)对其单糖组成及结构进行解析。结果:经分离纯化,SPS包含2个主要组分(SPS2及SPS3)。SPS2的相对分子质量为9.33×103,由鼠李糖、阿拉伯糖、木糖、甘露糖、葡萄糖、半乳糖6种单糖组成,其摩尔比为4.44∶1.46∶4.51∶5.82∶8.23∶19.38;SPS3的相对分子质量为5.86×103,由鼠李糖、阿拉伯糖、葡萄糖、半乳糖4种单糖组成,其摩尔比为2.93∶11.19∶33.68∶3.48。结论:SPS两组分都为均一杂多糖。

Objective： To determine the relative molecular weight（Mw） and monosaccharide compositions of safflower polysaccharides（SPS）.Methods： The polysaccharides were extracted from safflower with hot water,followed by precipitating with ethanol,freezing and drying,removing of free protein by the Sevag method and decolorizing with HPD-100,and finally separated into two main fractions SPS2 and SPS3 by DEAE-52 cellulose column and Sephadex G-100 gel column for chromatographic isolation and purification.The purity was identified by ultraviolet spectrum,the relative molecular weight（MW） was analyzed by high-performance gel permeation chromatography（HPGPC）,the monosaccharide compositions and their structures were investigated by derivative gas chromatography（GC） and infrared spectroscopy（FT-IR）.Results： The crude polysaccharide was fractioned to two main fractions（SPS2 and SPS3） by isolation and purification on DEAE-52 cellulose column and Sephadex G-100 column.SPS2,with Mw of 9.33 × 103,was composed of rhamnose（Rha）,arabinose（Ara）,xylose（Xyl）,mannose（Man）,glucose（Glc） and galactose（Gal） in a molar ratio of 4.44∶ 1.46∶ 4.51∶ 5.82∶ 8.23∶ 19.38.SPS3,with Mw of 5.86 × 103,was composed of rhamnose（Rha）,arabinose（Ara）,glucose（Glc） and galactose（Gal） in a molar ratio of 2.93∶ 11.19∶ 33.68∶ 3.48.Conclusion： The SPS compositions,SPS2 and SPS3,are both uniform heteropolysaccharide.