千斤拔多糖的提取纯化及其结构鉴定

利用正交试验考察千斤拔多糖的提取工艺,并比较脱蛋白方法中的Sevag法和三氯乙酸法的纯化效果,总糖含量测定采用苯酚-硫酸法,蛋白质含量测定采用考马斯亮蓝法;采用DEAE-52纤维柱法来分离多糖,并运用HPLC色谱来分析千斤拔多糖中的单糖成分。结果表明:经正交试验得出千斤拔多糖的最佳提取条件为时间2.5 h,料液比为1∶30,温度80℃,其多糖得率为8.558%。对比两种脱蛋白的方法,Sevage法萃取3次时蛋白的脱除效果最好。经DEAE-52纤维柱来分离多糖共分得7个组分。经HPLC色谱鉴定出有葡萄糖,甘露糖和阿拉伯糖,主要单糖成分为葡萄糖。

Extraction, purification and structural identification of Moghania polysaccharides

In order to investigate the extraction process of polysaccharide from the Moghania with the orthogonal design, we compared the deproteinization process conditions of polysaccharides by Sevage with TCA method, determined the total polysaccharide contents with the method of phenol-sulfuricacid and the protein content by the method of Coomassie bril-liant blue. And we also purified the Moghania polysaccharide by DEAE-52 and analyzed the monosaccharide composi-tionby of Moghania polysaccharide by the method of high performance liquid chromatography ( HPLC). The results showed that the best time, ratio of solid to liquid and extraction temperate of the extraction process were in 2. 5 h, 1: 30, 80℃, as the extraction condition, the yield of polysaccharide was 8. 558%. By comparison with the Sevage method and TCA method, the optimal conditions of deproteinization were proved to be extracted three times with Sevage method. Moghania polysaccharide was isolated into seven fractionates by DEAE-52 cellulose column chromatography; three kinds of saccharide were investigated by the HPLC, which were glucose, mannose and arabinose. In the three kinds of saccharide, peak area of the glucose was biggest than others, so the major saccharides was glucose.