Study On Extraction Techniques And Characterization Of Polysacharide From Aconitum Coreanum

Objective:To establish relations with polysaccharide purification process of white line, on the off white with polysaccharide-level components to make a comprehensive purification system, using advanced purification platform polysaccharide isolated uniform level copies. The simultaneous analysis of the physical and chemical properties of polysaccharides.Methods:Orthogonal design, access to off white with polysaccharide economic, efficient and complete extraction line. By repeated freezing and thawing, high-speed centrifuge and other conventional methods, and the protease law and Sevag method combined with removal of protein. Use of GE’s AKTA explore FPLC purification system, using ion exchange chromatography and molecular exclusion chromatography Combination, will carry different charges, different molecular weight polysaccharides separated from the complete system. At the same time, the use of Agilent 1100 preparative HPLC, with the use of Agilent ZORBAX GF-250 and GF-450 gel filtration column to ensure the purity of sample preparation. Analytical HPLC using polysaccharide components were identified once, and using GPC method to calculate the average molecular weight. By phenol-sulfuric acid method, Bradford method and UV-Vis spectrophotometer, the total sugar content, protein content and its spectrum characteristics were characterized by gas chromatography after acetylation determined the monosaccharide composition of each component and molar ratio. Integrated use of classical chemical analysis and instrumental analysis (GC, HPLC and IR) were used to study the basic structure of polysaccharides.Result The best extraction and purification process of polysaccharide in Aconitum coreanum were established. Result shows the optimum extraction temperature was 85℃, optimum time was 3 hr and liquid-to-solid ratio was 1:5. Under the optimum conditions, the yield of polysaccharide in Aconitum coreanum was 8.86%. The chemical components and the physical and chemical properties are researched. The results indicated that the polysaccharide of Aconitum coreanum was a acidic polysaccharide. Component sugars in a molar ratio was galactose(Gal):arabinose (Ara): xylose (Xyl):glucose (Glu):galacturnic acid=1:0.9:1.7:5:0.6 respectively.Conclusion:Optimizing the extraction process results in off white with significantly increased the yield of polysaccharide. Advanced polysaccharide purificantion platform ensure that all copies once. By ion exchange and gel filtration chromatography obtained the ACP-Ⅰ.