The Extraction And Separation Of HPS1, And The Study On Structural Characteristics And Anti-complementary Activity Of Four Components From HPS1

Four polysaccharides, HPS1-A, HPS1-B, HPS1-C and HPS1-D, were obtained by aqueous extraction followed by precipitation with ethanol and purified with anion-exchange on DEAE-52cellulose and gel-permeation chromatography on sephadex G-100from Hedysarum polybotrys Hand.-Mazz.In this paper we studied the structural features and anti-complementary activity of above-mentioned four fractions and Radix Hedysari polysaccharides1(HPS1). We compared the physicochemical properties of above-mentioned four fractions by the total sugar content, protein content, uronic acid content, elements composition, and molecular weight, monosacch-aride composition. In addition, we also compared the conformation of HPS1-A, HPS1-B, HPS1-C and HPS1-D by high performance gel permeation chromatography-multi angle laser light scattering (GPC-MALLS) and scanning electron microscopy(SEM).Then partial acid hydrolysis,uronic acid reduction, methylation, GC, GC-MS, IR, multi-angle laser light scattering and one-dimensional and two-dimensional nuclear magnetic resonancespectroscopy method were used to study chemical structural of HPS1-B and HPS1-D. The result showed that HPS1-B was a acidic polysaccharide, molecular weight was measured to be523.6KDa, which backbone was composed of a-L-Araf (1â†',â†'5) α-L-Araf (1â†'β-D-Glcp (1â†',â†'4)α-D-Glcp(1â†',â†'4,6) α-D-Glcp (1â†', residues substituted at0-6of galacturonopyranose. And that the side chains were composed ofâ†'4)α-D-GlapA(1â†',β-D-Glcp (1â†'andâ†'4) α-D-Glcp(1â†'), and occasionally by SO3-, and one sulfate ester occasionally occur per18-22residues. Its conformation has been studied by GPC-MALLS, the results showed that the conformation of HPSl-B has a random coil, and was mono-dispersed in0.9%NaCl solution.And HPS1-D proved to be a neutral sugar, withâ†'4) α-D-Glcp (lâ†',â†'4,6) α-D-Glcp (1â†'in backbone,and α-L-Araf (1â†',â†'5) α-L-Araf (1â†',â†'3,5) α-L-Araf (1â†'n branches. HPSl-D has a random coil state conformation with monodisperse mass distribution in0.9%NaCl solution.We used the classical pathway hemolysis to study anti-complement activity of HPS1, HPS1-A, HPS1-B, HPS1-CandHPS1-D.The results showed that HPS1-D, HPS1-A, HPS1-B anti-complement activity better, HPS1followed, HPS1-C activity is weakest.