红稗多糖的分离纯化工艺

Isolation and Purification of Polysaccharide from Bacca Sedge

为研究红稗粗多糖不同的脱蛋白工艺,并确定DEAE-52纤维素交换层析柱纯化分离红稗多糖的最优条件。用Sephadex G-100柱层析分离纯化得到的红稗多糖并采用柱前衍生化高效液相色谱法对纯化后的红稗精多糖的单糖组成种类进行测定。结果表明:红稗粗多糖的最佳上样浓度为5 mg/mL,洗脱速度0.5mL/min,上样体积25mL;采用Sevage法除蛋白3次,得到的蛋白清除率以及多糖保存率均较高,通过DEAE-52纤维素的柱层析分离纯化后可得到大组份多糖;再由Sephadex G-100柱层析分离纯化得到的大组份红稗中性多糖;再利用柱前衍生化高效液相色谱法(HCLP)测定分离纯化后的红稗多糖主要由L-鼠李糖(rhamnose,Rham)、D-葡萄糖醛酸(glucuronic acid,Glu)和葡萄糖(glucose,Glc)3种单糖组成。

In order to study deproteinization process of polysaccharide from bacca sedge with different methods,the optimum conditions of DEAE-52 cellulose exchange chromatography column were determined,then the purified polysaccharides were separated and purified by Sephadex G-100 column chromatography,and the monosaccharide composition of the purified polysaccharides was determined by pre-column derivatization HPLC.Results:The optimal sample concentration,elution rate and sample volume were respectively 5mg/mL,0.5mL/min and 25 mL.Protein removal rate and the polysaccharide storage rate were higher by sevage method.After separation and purification by DEAE-52 cellulose,a large polysaccharide was obtained by Sephadex G-100,then isolated and purified by column chromatography and high performance liquid chromatography(HCLP).The results showed that the purified polysaccharide was mainly composed of L-rhamnose(Rhamnose,Rham),Glucuronic acid(Glu)and glucose(Glc).