Study On The Extraction,Purification And Anti-Oxidation Activities Of Polysaccharide From Pine Cone Of Pinus Koraiensis

Pinus Koraiensis is a kind of pine genus plant and distributed in the north of east.Investigations indicated that polysaccharides had many biological activities such as anti-tumor and anti-virus.The topic used Pinus Koraiensis as raw materials,and studied on the extraction,isolation,purification and covered the anti-oxidation activities firstly.The related work were shown as follows.(1)A method to determine the contents of polysaccharides from pine cone of Pinus koraiensis was established-Improved-Diferential Phenol Sulfuric Method. This experiment was the first attempt.Compared with other Phenol Sulfuric Method,it can be used to determine the polysaccharides directly and overcome the errors caused by the addition of sulfuric acid.The topic studied on the method in the aspects of the wavelength,the time of hydrolysis and compound of the color liquid,the linear,the stability and reproducibility. The results showed that the method is simple, accurate,reproducible and can be used for the determination of Polysaccharides from pine cone of Pinus Koraiensis.(2)The polysaccharides from pine cone of Pinus Koraiensis was extracted by hot water and precipated by alcohol and the purification was studied.The experimental results showed that the polysaccharide yield was3.68%on the parameters involved decoction by hot water three times,ratio of water to material10:1,extraction time30min.Process of removing protein:By comparing the methods of protein removal including trichloroacetic acid,sevage and papain,the pain method was choosed as the optimal method finally.The optimum conditions were2.4ml of papain,pH5.5,temperature50℃,timel.5h.The amount of protein removed was56.83%while polysaccharide lossed8.33%during the deproteinization proccProcess of decolorization:Contrasting the methods such as activated carbon attaching method,hydrogen peroxide oxidation method,macroporous rosin adsorption method and ion exchange resion method, it was macroporous rosin adsorption process that had the most effect in decolorizing polysaccharide.The result is showed that D101resion was used to determine the optimum adsorption conditions for this method. The optimum conditions were2g of resion, pH5.0,temperature40℃,time5.0h.The decolorazation rate was72.16%while polysaccharide lossed28.03%(3) The integrated process route on the isolation and purification of polysaccharide was studied and the purity was detected.The crude polyszccharide was purification through papain added twice Sevage method to removal of proteins,ultrafiltration to removal the small molecules,the macroporous rosin adsorption process to decolorization,DEAE-52to elute by different solvent.Six components was obtained and three contents were higher.They were analysized through UV detection,freeze-thaw analysis and Biuret reaction.The result showed that the components were homogeneous components(4) The antioxidant activity of polysaccharides was studied through restore research,the clearing capability of hydroxyl radical as well as the DPPH.The results showed that polysaccharide from pine cone had a certain reduction capacity and it increased with the concentration of polysaccharide.In the case of the same concentration,the ability of polysaccharide was weaker than Vc, Purified polysaccharides scavenging ability was weaker than the crude polysaccharides.The reasons were being studied in the follow-up work.The topic studied on the analysis methods,extraction,isolation,purification and the anti-oxidation activities and achieved the obvious results.Most of them were reported firstly. The above studies can provide a theoretical basis and basic data for the rational development of pine cone.