海螵蛸多糖的提取分离及活性组分CPS-1的纯化  被引量:32

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

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作  者:魏江洲[1] 张建鹏[1] 刘军华[1] 王顺春[2] 冯伟华[1] 焦炳华[1] 

机构地区:[1]第二军医大学基础医学部生物化学与分子生物学教研室,上海200433 [2]上海中医药大学中药研究所,上海201203

出  处:《第二军医大学学报》2006年第1期17-21,共5页Academic Journal of Second Military Medical University

基  金:国家高技术研究发展计划/863计划(2004AA2Z3C30);浙江省科技计划项目(2005ZJKJJH2202).~~

摘  要: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.

关 键 词:海螵蛸 多糖类 分离 提纯 CPS-1 

分 类 号:R282.77[医药卫生—中药学]

 

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