天花粉多糖的除蛋白工艺优化研究

Optimization study of deproteinization process for polysaccharides from Trichosanthis Radix

目的:优化天花粉多糖的除蛋白工艺。方法:以除蛋白率、多糖收率和天花粉多糖中的中性糖、糖醛酸和蛋白质含量为指标,筛选并优化了天花粉多糖的除蛋白工艺。结果:天花粉多糖的最佳除蛋白工艺为提取液以50%乙醇浓度除杂,上清液调节乙醇浓度达80%醇沉;沉淀加水溶解,TCA浓度15%,体积1∶1,振荡30min;取上清液,再加入氯仿-正丁醇溶液体积比10∶1,剧烈振摇20min,萃取两次。除蛋白率为88.18%,多糖收率为75.07%,总糖含量可达77.74%,蛋白质含量仅为0.23%。结论:乙醇分级醇沉、TCA法和Sevage法的联合除蛋白质方法简便,成本较低,可大量除去天花粉多糖中的蛋白质和淀粉杂质,提高天花粉多糖纯度。

Objective：To optimize deproteinization process of polysaccharides from Trichosanthis Radix.Method：With retention rate of polysaccharides,deproteinized rate and the content of neutral sugar,uronic acid and protein as indexes,the optimized deproteinization process of polysaccharides from Trichosanthis Radix were studied.Result：The concentrated solution was precipitated sequentially by final concentration of 50%（v/v） ethanol and80%（v/v）ethanol,then the precipitate was removed protein by adding the equal volume of 15%TCA to deprotein30 min,and the last step is that the supernate was mixed with chloroform and n-butanol as volume ratio of 10：1 to severe vibrate 20 min and repeat two times.Retention rate of polysaccharides,deproteinized rate,the content of total sugar and protein were 75.07%,88.18%,77.74% and 0.23%,respectively.Conclusion：The deproteinized method united grade alcohol sink,TCA and Sevage method could improve the purity of polysaccharide from trichosanthis radix by removing the large amounts of protein and starch impurity,meanwhile the method is simple and low costing.