海螵蛸多糖的提取分离及活性组分CPS-1的纯化

Extraction of Cuttlebone polysaccharides and purification of their active component CPS-1

目的:从海洋中草药海螵蛸(乌贼骨)中提取分离得到多糖活性物质。对粗多糖中主要的活性组分CPS-1进行纯化,得到成分相对均一的天然活性多糖。方法:用热水抽提法提取海螵蛸多糖粗品,并对提取工艺进行正交设计,优化提取方法。对粗品组分进行总糖量测定。用DEAE-Sepharose F.F柱及Sepharose CL-6B柱对海螵蛸多糖粗品进行分离。通过活性检测实验确定活性部分。最后使用Sephacryl S-300柱进一步纯化。用HPLC来检测精品多糖的纯度。标准曲线法测定相对分子质量。结果:成功得到海螵蛸粗多糖。经过离子交换柱DEAE-Sepharose Fast Flow和Sepharose CL-6B柱及分子筛Sephacr-yl S-300柱的进一步分离纯化和活性跟踪,最后得到相对均一的活性精品多糖CPS-1,其含糖量达93.6%,相对分子质量为1×106。结论:不同的提取条件影响海螵蛸多糖的得率。不同性质的分离材料能够将相对分子质量及电荷有差异的多糖分离,达到分离的目的。精品多糖CPS-1是从海洋中草药海螵蛸中得到的均一多糖组分。

Objective：To extract the active polysaccharides from traditional Chinese herb cuttlebone and to purify cuttlebone polysacchride salts 1 （CPS-1）, an active component of crude polysaccharides, so as to obtain refined natural active polysaccharides. Methods： Hot-water extraction method was optimized by orthogonal designing and was used to extract crude polysaccharides from cuttlebone. The total sugar contents of crude polysaccharides were determined. DEAE Sepharose F. F column and Sepharose CL-6B column were applied to separate CPS-1 from the crude polysaccharides. The active components were determined by animal experiments and Sephacryl S-300 column was used for further purification of CPS-1. HPI.C was used to determine the purity of refined CPS-1 and the molecular weight of CPS- 1 was determined by polysaccharides standard curve. Results： Crude polysaccharides were successfully extracted from cuttlebone. After purification with DEAE Sepharose F. F column, Seph arose CL-6B column and Sephacryl S 300 column and activity study, refined CPS-1 with average molecular weight （1×10^6 ） was obtained,and the sugar content reached 93.6 % . Conclusion： Different extraction conditions have different extraction results for crude polysaccharides and different separation materials can separate the mixed polysaccharides by their molecular weights or charge characteristics. CPS- 1 is a natural active polysaccharide extracted from cuttlebone.