Study On Isolation,Purification,Structural Identification And Its Bioactivities Of Polysaccharide From Arenaria Kansuensis Maxim

The water extraction process and alkali extraction process of polysaccharides from Arenaria kansuensis Maxim was studied in this paper.;And isolate and purify the crude water extraction polysaccharides of Arenaria kansuensis Maxim;Then to analyze the structures of its purity;Use the simulation experiments in vitro and animal experiments to determine the effects of different types of Arenaria kansuensis Maxim polysaccharides on the effects on scavenging active oxygen free radicals and the ability of hyprozia tolerance.The main results were shown as follows:1.Two different extraction process of polysaccharides from Arenaria kansuensis Maxim and the optimization of the two processTo study the water extraction process and alkali extraction process of polysaccharides from Arenaria kansuensis Maxim respectively.The water extraction process use the quadratic regression rotational combination design to determine the effects of extraction temperature,extraction time,extraction proportion of solvent and stuff on the extraction yields of polysaccharides from Arenaria kansuensis Maxim.Use the extraction yields of polysaccharides as the response value in order to set up a regression equation and the regression models.The regression equation was as follow:Y=8.169492+1.1985^*X₁+0.55475^*X₂+0.69725^*X₃-0.264944^*X₁^*X₁-0.12675^*X₂^*X ₁-0.136194^*X₂^*X₂-0.095^*X₃^*X₁-0.0715^*X₃^*X₂-0.175819^*X₃^*X₃. After optimizing the combination factors by computer determined the optimum extraction parameters were:temperature(94.5℃-97.5℃);time(2h-2.5h);extraction proportion of solvent and stuff(43v/w-51v/w).Alkali extraction process of polysaccharides from Arenaria kansuensis Maxim was studied on the base of one-way anova test,through orthogonal test determined the optimum extraction parameters of alkali extraction process of polysaccharides from Arenaria kansuensis Maxim were:temperature 95℃,time 1.5h,extraction proportion of solvent and stuff 30:1 v/w,NaOH concentration 2.5 mol/L.2.Isolation and purification of polysaccharide of water extraction from Arenaria kansuensis MaximDEAE-SephadexA-25 chromatography and Sephadex G-75 gel column chromatography was used for the isolation and purification of the cure polysaccharide of water extraction from Arenaria kansuensis Maxim.The results showed that:after cure polysaccharide of water extraction from Arenaria kansuensis Maxim which has already elimination impurity was isolated and purified by DEAE-SephadexA-25 chromatography, five components AMP1,AMP2,AMP3,AMP4,AMP5 were got,the yield of them were 26.53%,5.77%,9.31%,18.32%,7.41%respctively;AMP1-B and AMP4-A the yield of which were 32.32%and 27.17%respectively,were purified from AMP-1 and AMP-4 through the Sephadex G-75 gel chromatography.3.The analysis of the primary structures of AMP1-B,AMP4-AThe analysis of the primary structure of the Arenaria kansuensis Maxim polysaccharides was studied through infrared spectroscopy,UV scanning analysis,gas chromatography. The results showed that:AMP1-B and AMP4-A was composed of rhamnose,arabinose, xylose,mannose,glucose and galactose as the common components,with the ratio of Rha:Ara:Xyl:Man:Glu:Gal=0.27:0.20:0.29:1.0:0.85:0.73 and 0.37:0.4: 0.41:1.0:0.68:0.86 in molar ratio respectively.AMP1-B and AMP4-A was provided with the absorption peak of characteristics of polysaccharide,a few of sugary protein, longer side chains and more branches and O-glucosedes,but without helix form.4.Antioxidant in vitro and the effect of the ability of hyprozia tolerance of Arenaria kansuensis Maxim polysaccharidesThrough simulation experiments in vitro,we researched the effects of different types of Arenaria kansuensis Maxim polysaccharides on hydroxyl radical and DPPH radical;the effect of the ability of hyprozia tolerance of different types of Arenaria kansuensis Maxim polysaccharides were researched by animal experiment,and then choosen the best one to study its dose-effect relationships.The results showed that:AAMP and WAMP can both effectively inhibited the activity of hydroxyl radical and DPPH radical.AMP can enhance the ability of hyprozia tolerance of rats obviously.Mid-dase AMP can increase the ability of hyprozia tolerance of rats most enfficency,and alleviate the hypoxic injury better.