Study On Structure Analysis And Anticoagulant Function In Vitro Of Garlic Polysaccharides

The human health was seriously threatened with thrombus embolism disease. At pre-sent, anticoagulant medicine was mostly used for clinic treatment was heparin, which had some side-effects, such as causing platelet disease, hemorrhage disease, hyperkalemia and so on. So it was essential to develop new type of medicine or functional food.Garlic not only had nutrition, but also had pharmacological effects. Garlic polysac-charide was one of the important effective components in garlic, which included allin and various physiological activities. Garlic polysaccharide which was rich in garlic had anti-oxidation, antiviral, enhanced immunity, protected liver and other pharmacological effects, making garlic polysaccharide have important development value. The anticoagulant func-tion in vitro of crude garlic polysaccharide which extracted by different methods were analyzed, the crude polysaccharide was separated from garlic with alkaline, the optimum extraction condition were determined by means of singer factor and orthogonal experi-ments, three components of garlic polysaccharides (GPⅠ, GPⅡ, GPⅢ) were purified from garlic by DEAE-52 cellulose column, the purity of GPⅠ, GPⅡ, GPⅢwere verified by Sephadex G-100 and frozen-thawing-analysis, the monosaccharide constituents and structure of three components of garlic polysaccharides were determined by gas chroma-togram and IR, respectively. GPC-MALLS-RI was applied to determine molecular struc-ture of GPⅠ.The results showed that:The crude garlic polysaccharide had anticoagulant activity, which worked by affecting intrinsic pathway coagulation systerm to prolong APTT of human plasma effectively. There were differences among crude garlic polysaccharides which extracted by different methods, the garlic polysaccharide which extracted by alkaline had the strongest antico-agulant activity.The parameter of alkali extract technology was perfected: the extraction temperature was 55℃, the extraction time was 270 min, the ratio of material to solvent was 1:8, the concentration of NaOH was 0.5 mol/L, respectively. Under these conditions, the extraction rate of crude garlic polysaccharide was 15.6%.The anticoagulant activity of garlic polysaccharide strengthened by deprotein. In ex-perimental area, the higher concentration of garlic polysaccharide, the stronger anticoagu-lant activity. Three polysaccharide components (GPⅠ, GPⅡ, GPⅢ) were separated by DEAE-52 cellulose column, which occupies of total garlic polysaccharides were 75.2%, 15.5%, 9.3%, respectively. There were no obvious difference among three components of garlic poly-saccharides on anticoagulant activity.Gas chromatogram analysis indicated that GPⅠcontained rhamnose, fructose, galac-tose, glucose and mannose and one unknown monosaccharide, with the constituent ratio of 15.4%:6.9%:19.7%:5.9%:30.5%:21.6%; GPⅡcontained was composed of rhamnose, fructose, galactose, glucose and mannoseand unknown nmonosaccharide, with the con-stituent ratio of 36.4%:11.9%:17.1%:9.6%:24.9%; GPⅡcontained fructose, galactose, glucose, mannose and one unknown monosaccharide, with the constituent ratio of 25.6%:22.5%:13.7%:15.1%:23.1%.Infrared spectrum analysis showed that three three components of garlic polysaccha-rides all have the typical character of polysaccharides. GPⅡand GPⅢappeared absorp-tion peak at 835 cm-1 and 831 cm-1, respectively., which suggested they containedα-glycosidic bond.GPC-MALLS-RI determined that the molecular weight of GPⅠrange from 5.2×104 g/mol to 5.6×104 g/mol, the root mean square (rms) radius range from 31.5 nm to 32.9 nm, two polydispersity values are 1.039 and 1.084, respectively. It was spherical polymer. The results showed that the garlic polysaccharide have potential development value.