| Vaccinium uliginosum belongs to Ericaceae Vaccinium genera. The ripe fr uit contains many elements such as polysaccharose, anthocyanin and flavone. According to researches, the fruit of Vaccinium uliginosum have many physical health-care functions like facilitating the resynthesis of erythropsin, anti inflammation, angiocardiopathy, Senescence, cancer and the function of improving immunity. Also, the Vaccinium uliginosum posses the biological activity of Antioxidant, anti-aging and regulating immune function; thus it plays an important role in clinical application and economic development. In order to deepen the development of Vaccinium uliginosum, this paper studies the extraction, purification, its structure and activity. In this way, it can provide a theoretical basis for the further study of Vaccinium uliginosum.Pressurized solvent extraction was used to extract polysaccharide, and the single-factor analysis was first conducted. Based on this experiment, the Central Composite Design was used to study the four factors which influences the yield of polysaccharide, including liquid-solid ratio, extraction temperature, time and pressure. The optimum extracting procedure was adopted through the response surface analysis as follows: extraction temperature 8 1 °C, extraction time 23 min, pressure 1.2 MPa, the yield of polysaccharide was 7.57%. The validation test polysaccharide yield is 7.48± 0.11%, close to forecast.The method of polyamide column is used for decolorizing polysaccharide of Vaccinium uliginosum fruit. The decolorization rate is 62.54±1.27%; protein removal rate reach ed 89.87±0.85%. the recovery rate of polysaccharide is 50.47±0.97%. The result of purity VUP-? could reach 55.65±0.39%.Purified polysaccharide was purified under the optimal conditions through Sephadex G-100: sample concentration 17 mg/m L, deionized water as eluent, maximum sample volume 0.8 m L, optimal elution rate 1m L/min. After the experiment, two main polysaccharides was obtained; the purity of VUP-?-1 was 83.35±0.32%; the purity of VUP-?-2 was 74.94±0.17%.Physicochemical properties and structural characteristics of VUP-?-1 were studied. The surface of VUP-?-1was gray in color, it was soluble in water, was insoluble in alcohol, ether, chloroform, acetone, butanol and other organic solvents. It does not contain substances such as proteins, reducing sugar, starch and phenolic. The content of uronic acid was determined by carbazole- sulphuric acid method, the value reach 20.34±1.02%.By HPLC purity identification, VUP-?-1 sample had weight average molecular weight Mw of 4.98×104 ku, Number average molecular weight Mn of 3.23×104 ku, molecular weight distribution index ?(Mw/Mn) of 1.54. Applying gas chromatographic to analyze monosaccharide composition of polysaccharides VUP-?-1 was studied. VUP-?-1 was composed of arabinose and galactose with molar ratio of 5:2. IR spectrum showed VUP-?-1 might have characteristic absorption peaks of glycosidic bonds of pyrylium. UV spectroscopic examination revealed that the polysaccharides were protein-free, nucleic acid free, and polyphenols-free.The surface of VUP-?-1 was exanimated by SEM scanning, it had a chain structure with branches and flexibility, at the same time, it also was aggregates with mutual staggered annularsugar chain, and there was a strong interaction between molecules. Congo red experimental result showed there was triple helix structure from VUP-?-1.According to the antioxidant research towards VUP-?-1, it had radical scavenging activities on DPPH·,·OH and O2-·. The scavenging rate rises with the concentration. When the concentration reached 1.2mg/m L, the scavenging rate were 93.86±0.59% ? 11.68±0.34% ? 5.50±0.83% respectively. The recovery power of VUP-?-1 was positively related with its concentration. It showed strong recovery power within the concentration range from 0 to 1.2mg/m L. When the concentration was 1.2mg/m L, VUP-?-1 showed strong reducing power which was 1.14±0.05.The alpha amylase removal experiment showed that VUP-?-1 can impose inhibition on alpha amylase; also, in a certain concentration range, when the concentration was 4mg/m L, the inhibition can reach to 37.23±0.45%.VUP-?-1 showed obviously inhibitory effects on the formation of dicarbonyl compounds and AGEs. Inhibition rates were 42.10±0.61% and 45.93±1.27% respectively, stronger than that of amadori compounds 29.91±0.84%, indicating that VUP-?-1 inhibit the glycation mainly occurred in the last two periods. |
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