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目的:对六味地黄生物制剂粗多糖中蛋白质的去除方法进行比较研究,探讨不同方法对多糖的提取率及纯度的影响。方法:利用Sevage法、TCA-正丁醇沉淀法、鞣酸沉淀法对六味地黄生物制剂脱蛋白,通过蛋白清除率和多糖保存率比较3种方法的优劣。结果:TCA-正丁醇法脱蛋白时效果最好,最佳条件为TCA-正丁醇试剂的体积比1∶2,TCA-正丁醇试剂的配比1∶5,振摇时间30 min,静置时间1.0 h,多糖脱蛋白效率最高,多糖保存率最大。结论:TCA-正丁醇法是目前六味地黄生物制剂脱蛋白效果最好的工艺。
OBJECTIVE: To compare the protein removal methods of crude polysaccharides from Liuwei Rehmanniae Preparata, and to explore the influence of different methods on the extraction rate and purity of polysaccharides. Methods: Sevage method, TCA-n-butanol precipitation method and tannic acid precipitation method were used to deproteinize the Liuwei Rehmannia glutinosae biological preparation. The advantages and disadvantages of the three methods were compared by the protein clearance rate and the polysaccharide preservation rate. Results: TCA-n-butanol deproteinization had the best effect. The best conditions were the volume ratio of TCA-n-butanol reagent 1: 2, the ratio of TCA-n-butanol reagent 1: 5, shaking time 30 min , Standing time 1.0 h, the highest efficiency of polysaccharide deproteinization, polysaccharide preservation rate. Conclusion: The TCA-n-butanol method is the best technology for deproteinization of Liuwei Rehmanniae Biologics.