Isolation, Purification Of Polysaccharides From Chinese Pine Pollen And Its Antioxidant Function

Chinese pine pollen polysaccharides is well-known for its bioactive function such as anticancer, enhancing immunity, et al. The Chinese pine pollen was produced in Chengde. The extraction conditions were inspected. The optimum way of coloration in phenol-sulfuric acid method was investigated. The isolation and purification were carried by DEAE-cellulose and Sephacryl S-200chromatography. The identification of purity was implied with Sephacryl S-300chromatography. Antioxidant abilities of crude and refining Chinese pine pollen polysaccharides were measured as activity against hydroxyl radical and superoxide anion-scavenging activity. By major work, access to the conclusions include the following:(1) Using only glucose as the standard, the optimum way of coloration in the process of determining Chinese pine pollen polysaccharides content by phenol-sulfuric acid method was obtained as follows:1.0mL5%phenol solution was added to2.0mL pollen polysaccharides solution and mixed. Then6.0mL concentrated sulfuric acid was added quickly drop by drop. After setted at100℃for10minute, the solution was detected at wavelength of483nm.(2) The extraction of polysaccharides by water from Chinese pine pollen was studied. The effects of four factors such as temperature, time, ratio of solid-liquid and times of extraction were investigated. The results showed that the effect of temperature was the most important, the ratio of solid-liquid was the second, times were the third, and time was the least. Through orthogonal test, the optimum conditions of extraction were obtained on the basis of single factor test. The results were as follows:temperature of70℃, ratio of liquid to solid (w/v) of1:12,2hours twice. In this condition, the content of polysaccharides in the extracts was1.81%.(3) The primary purification of the refining Chinese pine pollen polysaccharides were carried out by DEAE-cellulose chromatography. Polysaccharides D1and polysaccharides D3were mainly obtained in this process. The polysaccharides D11and polysaecharidesD31were achieved by further purifying D1and D3respectively with Sephacryl S-200chromatography. The identification of purification with Sephacryl S-300chromatography implied that D11and D31were homogeneous polysaccharides. The molecular weight of D11was76468dalton, estimated by gel filtration. The monosaccharide compositions of polysaccharides D1were analyzed by gas chromatography. No typical absorption of nucleic acid or Protein in Polysaccharides D11was observed in280nm of UV spectrum. Infrared spectrum of polysaccharides D11indicates that they had typical characteristic absorption peaks of polysaccharides.(4) Antioxidant abilities of crude and refining Chinese pine pollen polysaccharides were measured as activity against hydroxyl radical and superoxide anion-scavenging activity. The investigation demonstrated that they possessed free radical scavenging activity. The ability of scavenging was enhanced with the concentration increases. And the ability of scavenging of the refining polysaccharides is better than the crude polysaccharides.