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目的通过对党参多糖(CPP)的提取纯化工艺优化及其单糖组成和相对分子质量(M)分布进行研究,为进一步分离CPP提供依据。方法采用硫酸-苯酚法测定多糖的量,采用正交设计确定最佳提取工艺;对提取的粗多糖进行脱蛋白、脱色、透析、冷冻干燥处理,得到精制粗多糖,并采用HPLC和高效凝胶色谱(HPGPC)法,对多糖水解后的单糖组成和相对分子质量分布进行测定。结果 CPP的最佳提取工艺为提取温度85℃,料液比1∶12,提取次数2次,提取时间1.5 h。在上述条件下,CPP的提取率可达到22.57%。水解单糖组成为葡萄糖醛酸、氨基半乳糖、木糖和少量甘露糖,多糖重均相对分子质量(Mw)为21 498。结论为CPP的分级和活性研究提供了理论基础。
OBJECTIVE: To optimize the extraction and purification process of Codonopsis pilosula polysaccharide (CPP) and its monosaccharide composition and relative molecular mass (M) distribution, and to provide a basis for further separation of CPP. Methods The amount of polysaccharides was determined by sulfuric acid-phenol method. The optimum extraction process was determined by orthogonal design. The crude polysaccharides were deproteinized, decolorized, dialyzed and freeze-dried to obtain crude polysaccharides. HPLC and high performance gel Chromatography (HPGPC) method was used to determine the monosaccharide composition and relative molecular mass distribution of polysaccharides hydrolyzed. Results The optimal extraction process of CPP was extraction temperature 85 ℃, solid-liquid ratio 1:12, extraction times 2 times and extraction time 1.5 h. Under the above conditions, the extraction rate of CPP can reach 22.57%. Hydrolyzed monosaccharides were composed of glucuronic acid, galactosamine, xylose and a small amount of mannose. The weight average molecular weight (Mw) of polysaccharides was 21 498. Conclusion provides a theoretical basis for the study of grading and activity of CPP.