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

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.HPLC 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,Sepharose 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.