Studies On The Polysaccharide Of Anemone Raddeana Regel

Purpose: Through the study of the Anemone raddean Regel polysaccharide, for further study of its chemical structure, pharmacology, biological activity and provide a basis for application development.Method:By orthogonal experiment,the optimum extraction condition of polysaccharide from Anemone raddeana Regel is studied. The two methods of Sevage method and Enzyme combined with Sevage method were used to remove proteins from the crude polysaccharides in Anemone raddeana Regel, to find the mosteffective extraction way for the Anemone raddeana Regel polysaccharides,several decolor methods of the polysaccharides were extimated. Five fractions designatedAP-Ⅰa , AP-Ⅱa～AP-Ⅱd from Anemone raddean Regel were isolated by dialysis ,gradient ethanol precipitation and activated carbon column chromatography. Using GC on the part of monosaccharide composition of species were tested. The antineoplasmic activity of polysaccharide samples on Hela cell was by MTT bioassay invitro. Result: According to the results of orthogonal experiment, the ratio of the sample weight to the solvent volume is 1:20,the temperature is 100℃, the duration is 2h, the concentration ratio 2.80% alcohol precipitating for 12h.. By comparing the effects,it was found that Enzyme combined with Sevage method,the polysaccharide was higher. to find the mosteffective extraction way for the polysaccharides of Anemone raddeana Regel.The result showed that it is an effective decolor way using macroporous resin D941 exchange column with H2O. Five fractions designatedAP-Ⅰa , AP-Ⅱa～AP-Ⅱd from Anemone raddean Regel were isolated by dialysis ,gradient ethanol precipitation and activated carbon column chromatography. The yields of them were 86.79%,39.22%,39.98%,42.27%,41.08%,and the Sugar hyaluronic acid content were10.90%,7.38%,6.85%,8.61%,10.21%。The GC analyses show that AP-Ⅰa and AP-Ⅱc were composed of rhamnose, arabinose, and glucose; AP-Ⅱa was composed of rhamnose, arabinose, glucose and galactose; AP-Ⅱd was composed of rhamnose, fucose, glucose and fucose.By IR spectrum, AP-Ⅰa was identified containing theαand theβ-pyranoid ring.In the end,the antineoplasmic activity of polysaccharide samples on Hela cell was by MTT bioassay in vitro.The results showed that all samples had apparent antineoplasmic activity on Hela cell,the antitumor effects will strengthen with the increasing concentration AP-Ⅰa and AP-Ⅱc.Conclusion: Polysaccharide of Anemone raddean Regel obtained by orthogonal test the best extraction process, and to determine the enzyme-Sevage and D941 macroporous resin made for a better purification method. Five fractions designated AP-Ⅰa , AP-Ⅱa～AP-Ⅱd from Anemone raddean Regel were isolated,the results showed that the MTT method: AP-Ⅰa and AP-Ⅱc have anti-tumor activity. These results, in order to further study the chemical structure of polysaccharides, pharmacology, biological activity and laid the foundation for application development.