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Study on Extraction, Purification and Biological Activities of Polysaccharides from Apple Snail (Ampullarum Crossean)

Author: XuGuiQin
Tutor: ChenQiang；HuangZhiJian
School: Fujian Agriculture and Forestry University
Course: Basic Veterinary Science
Keywords: Apple snails polysaccharide Extract Purification Structural Analysis Biological activity
CLC: R284
Type: Master's thesis
Year: 2008
Downloads: 182
Quote: 2
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Abstract


Objective: medicinal value for in-depth study of animal polysaccharide, apple snails polysaccharide separation and purification, structural identification and biological activity. Method: 1. Extracted by hot water extraction and alcohol precipitation apple snails polysaccharide in the univariate analysis (extraction time, extraction times, solid-liquid ratio, extraction temperature) on the basis of the L 9 4 orthogonal to optimize the extraction conditions. 2 apple snails polysaccharides After addition was purified protein impurities, cellulose ion exchange column (DEAE-52), and dextran gel columns (SephadexG 200), by Ultraviolet Spectroscopy (UV) and high performance liquid chromatography (HPLC) analysis of the purity of the polysaccharide. 3 combined with HPLC, gas chromatography (GC), infrared spectroscopy (IR), thin layer chromatography (TLC) methods and chemical analysis preliminary identification of the primary structure of the polysaccharide and some physicochemical properties. 4 by in vitro antioxidant in vitro DNA protective effect of immune suppression and in vivo mouse model of immune function tests, analysis of activity of polysaccharide oxidation and improve immunity. Results: 1. Orthogonal test showed that the optimal extraction of polysaccharide: a temperature of 90 ° C, solid-liquid ratio of 1:90, the time for 2h. Phenol - sulfuric acid method determination of the polysaccharide content the final measured the apple snails polysaccharide sugar content of 39.88%. DEAE-52 ion-exchange column and Sephadex Sephadex G-200 column the apple snails polysaccharides were isolated and purified, ASPS-I a and ASPS-II a two polysaccharides component. By UV spectroscopy at 260nm and 280 nm, no nucleic acid and protein absorption peak. By HPLC, ASPS-I a have an absorption peak, ASPS-I a homogeneous polysaccharide, its composition and structure analysis. 3.ASPS-I a thin-layer chromatography, infrared spectroscopy, high performance liquid chromatography and gas chromatography analysis showed that the characteristic absorption peak of the polysaccharide composed of glucose alone, ASPS-I a main chain alpha-pyran The glycosidic linkage predominating polysaccharide. 4 antioxidant in vitro experiments showed that the the apple snails polysaccharide has good resistance to oxidation, as follows: the polysaccharide concentration 3.0 mg / mL when the hydroxyl radical on O 2 - < / sup> the highest clearance rate reached 75.07% and 42.41%, respectively, in the polysaccharide concentration of 5 mg / mL, H 2 O 2 -induced hemolysis of erythrocytes inhibition rate reached 56.7%, anti-liver tissue of spontaneous lipid oxidation inhibition rate reached 67.33%. The apple snails polysaccharide also has a protective effect on the strand of DNA caused by free radicals and DNA base damage. The in vitro and in vivo immune tests showed that the apple snails polysaccharide can enhance macrophage phagocytic capacity, increased spleen index, enhanced activity of mice, to improve all aspects of immune activity in mice. Conclusion: water extraction and alcohol precipitation method to extract apple snails polysaccharide, the best extraction process. The purified component is relatively uniformly in the main chain alpha-pyran-glycosidic bond is connected to the main, a polysaccharide composed of glucose single; the apple snails polysaccharide having good resistance to oxidation and enhanced immune activity.

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