油茶籽多糖分离纯化和结构分析

Study on the purification and construction of polysaccharides from Camellia oleifera

以120min、50℃、1:3的料液比提取多糖粗品,并采用Sevage法对得到的油茶籽多糖粗品进行脱蛋白,再通过DEAE-纤维素、SephadexG-100柱层析进一步纯化,得到三种类型(CPSⅠ、CPSⅡ、CPSⅢ)的油茶籽多糖。油茶籽多糖样品的水溶液在紫外-可见光260nm(核酸)和280nm(蛋白质)处无明显吸收,但在190nm到210nm之间有多糖特征吸收峰。通过柱层析标准样品分析,可以得出CPSⅠ的分子量大约为256035Da,CPSⅡ的分子量大约为10290.3Da,CPSⅢ的分子量大约为3590Da。薄层色谱分析初步推断CPSⅠ由半乳糖和木糖组成;CPSⅡ由果糖和木糖,以及另一种不明的单糖组成;CPSⅢ由半乳糖组成。通过红外光谱分析推测CPSI可能是含有磷酸基团的多糖;CPSII和CPSIII都含有纯的醇基团,且CPSII中的一元醇、二元醇的含量比CPSIII高,CPSII和CPSIII都含有α糖苷键和β糖苷键。

The crude polysaccharides which were extracted under the condition of 120min of the extraction time,50℃ of the extraction temperature,the 1∶3 ratio of material to liquid,and camellia oil was received for crude polysaccharide from protein by the Sevage method,DEAE-cellulose column,SephadexG-100 column chromatography for further purification of polysaccharides camellia oil.The results showed that the Camellia oleifera polysaccharides was composed of three types of polysaccharides（CPSⅠ,CPSⅡ,CPSⅢ）.What＇s more,by ultraviolet-visible spectrum between 190nm and 210nm,there was a Absorbance of polysaccharides＇ features between 190nm and 210nm,not 260nm point and 280nm point,which was two Absorbability peaks of nucleic acids＇ and proteins＇ features respectively.Through the thin-layer chromatography,we can initially infer that CPSⅠ was composed of galactose and xylose,CPSⅡ was composed of fructose and xylose,as well as another unidentified monosaccharide composition,and CPS Ⅲ was composed by galactose.And through standard samples by column chromatography,CPSⅠ molecular weight was about 256035Da,CPSⅡ was about 10290.3Da,CPS Ⅲ was about 3590Da.Through infrared spectral analysis,CPSI speculate may contain phosphoric acid group of polysaccharide,CPSII and CPSIII contain pure alcohol based corporations,and CPSII contain more unitary alcohol,binary alcohol than the CPSIII.Both CPSII and CPSIII contain α polysaccharide glucoside bond and β glucoside bond.