Sevag法脱鹿药多糖蛋白工艺条件优化

Study on the orthogonal experiment design of removing protein from polysaccharide of Smilacina japonica

关键词:  鹿药  多糖  Sevag法  脱蛋白  工艺优化

Key words:  Smilacina japonica  polysaccharide  Sevag method  removing protein  process optimization

摘要:

为了优化Sevag法脱鹿药粗多糖蛋白工艺条件。以牛血清白蛋白为标准品测定蛋白质含量，以葡萄糖为标准品测定多糖含量，以蛋白质脱除率、多糖保留率及综合评分为指标，通过正交试验考察氯仿与正丁醇的比例、多糖液与Sevag液的比例、振荡时间、洗脱次数四因素对脱蛋白效果的影响，确定鹿药多糖脱蛋白的最佳工艺条件。结果表明Sevag法脱鹿药多糖蛋白的最佳条件为氯仿:正丁醇=4:1，多糖溶液（2 mg/mL）:Sevag试剂=2:1，振荡时间15 min，洗脱6次，从综合评分来看，主要的影响因素是脱蛋白次数和振荡时间。Sevag法脱鹿药多糖蛋白的方法简便，工艺稳定且可有效提高多糖的纯度。

Abstract:

To optimize the technology of removing protein from crude polysaccharide of Smilacina japonica. The Sevag method was used remove the protein, bovine serum albumin as the standard test protein content, glucose as the standard test polysaccharide content. The effect of removing protein according to the index of the removal protein rate, the polysaccharide retention rate and comprehensive score, through orthogonal experiment study the ratio of chloroform and n-butyl alcohol, the ratio of polysaccharide solution and Sevag solution, oscillation time, elution times and so on. The optimizing parameters for the removing protein of Smilacina japonica polysaccharide were chloroform: butanol =4:1, polysaccharide solution: Sevag reagent =2:1, 6 times, 15 min of each time. According to the comprehensive evaluation, the significant factors are the oscillation time and the frequency. This optimum technology of Sevag method to removing protein is stable, and the method is simple and effective. It can improve the purity of polysaccharide.