油茶籽饼粕多糖的提取工艺优化及结构鉴定

Optimization of Extraction Process and Structure Analysis of Polysaccharide Extracted from Camellia Oleifera Seed Cake

[目的]以油茶籽饼粕为原料,优化油茶籽饼粕多糖提取工艺,并对纯化后的多糖进行结构鉴定。[方法]根据单因素试验结果,以超声时间10、15、20 min;料液比1∶15、1∶20、1∶25;浸提温度50、60、70℃;浸提时间60、90、120 min为影响因素,多糖得率为指标,通过正交试验设计优化提取工艺。粗多糖经脱色,脱蛋白处理后,再经过DEAE-52纤维素阴离子交换树脂纯化,并对纯化多糖组分进行紫外光谱和高效液相色谱分析。[结果]超声时间10 min,料液比1∶20,浸提温度60℃,浸提时间120 min为最佳工艺条件,此条件下油茶籽饼粕多糖的平均得率16.76%。粗多糖经脱色、脱蛋白、纤维素阴离子交换树脂柱层析得到纯化组分CCP。紫外光谱表明CCP为较纯的均一多糖组分,高效液相色谱表明CCP的单糖组成为甘露糖和葡萄糖,其物质的量比为:1.91∶1.00。同时还含有少量的鼠李糖。[结论]该工艺优化合理,多糖的结构得到了初步的确证。研究为油茶籽饼粕多糖的后续理化活性分析、结构表征提供了理论依据。

[Objective]Camellia oleifera seed cake was used as raw material to optimize the extraction process of polysaccharides and to analyze the structure of purified polysaccharide.[Method]Ultrasonic time(10,15,20 min),ratio of material to liquid(1∶15,1∶20,1∶25),extraction temperature(50,60,70℃),extraction time(60,90,120 min)were choosen as influencing factors on the basis of single factor experiments,extraction rate of polysaccharides was taken as evaluation indexe.Orthogonal test was used to optimize polysaccharide extraction process.The crude polysaccharides was decolorized,deproteinized and purified by DEAE-52 Cellulose anion exchange chromatography respectively.The purified polysaccharides were analyzed by UV and high performance liquid chromatography(HPLC).[Result]The best extraction process was ultrasonic time 10 min,ratio of material to liquid 1∶20,extraction temperature 60℃,extraction time 120 min.Under these conditions,the average extraction rate of polysaccharides reached 16.76%.Purified component was obtained and named as CCP.UV spectrum showed that CCP was a homogeneous polysaccharide component.HPLC showed that the purified polysaccharide was mainly composed of mannose,glucose with molar ratio of 1.91∶1.00,and tiny amounts of rhamnose.[Conclusion]The optimized process is reasonable and the structure of polysaccharides is preliminary identificated.It can provide a theoretical basis for further activity analysis and structural characterization of camellia oleifera seed cake polysaccharides.