Study On Separation And Purification Of Polysaccharides From Gingko Biloba Sarcotesta And Their Biological Activities

Gingko is one of the rarely medical plant that its leaf,fruit and sarcotesta all have medical development value.It is called living fossil that all over body is precious.According to the statistics,the Gingko plant spread all over 27 province (city)in our country at present .It can produce gingko semen 1.2 thousand ton and the gingko biloba sarcotesta account 70% of the whole gingko fructification.According to this,there are 3thousand ton fresh gingko biloba sarcotesta.We can get 1.2 thousand dried gingko biloba sarcotesta after remove 60% water. Gingko leaf extraction and its preparation is one of the natural medicine that welcomed to people at present.Various kinds of nutrition food that developed by Gingko is fairly.Only the development of Gingko Biloba sarcotesta is lower.Polysaccharide is one of the important biological activity component.The gingko biloba sarcotesta is abundance in our province.It is reported that the water extracted polysaccharides of gingko biloba sarcotesta have the activity of anti-tumor, immunoloregulation,anti-aging,and so on.At present,gingko biloba sarcotesta is the rejection of agriculture.If we can develop its polysaccharides to be intermediate product of medicine or health food ,the gingko resource will be utilized.Therefore,in this paper,using gingko biloba sarcotesta as test material,the total polysaccharides in this material were analyzed,which its extraction,separation and purification analysis of qualitative and quantitative were deeply studied.At the same time,we studied the immunoloregulation activity of crude and deproteinization polysaccharides and anti-tummor activity of crude, purification and acetylated polysaccharides.The main contents and conclusions are as follows:1.To study the extraction conditions of polysaccharides by using water.The conditions of extraction were optimized by orthogonal experiment on the basis of singal factor tests.The result showed that the optimal conditions was as follows: The extraction temperature is 100℃, the time is 1h ,and the solid-liquid ratio isl:12 .The effect order of three factors for the extraction temperature, time,solid-liquid ratio,and in turn. 2.To study the condition of determination on polysaccharides from gingko biloba sarcotesta by using phenol-sulfuric method.The conditions of determination were optimized by orthogonal experiment on the basis of single factor tests.The result showed that the optimal conditions were as follows: The dosis of 5% phenol is 1.2mL, the dosis of sulphuric acid is 4mL ,temperature is 30℃and time is 20min. The average recovery was 99.60%,and the relative standard deviation was 3.02%(n=5).3.To study the immunoloregulation activity of crude(GBEP-1) and deproteinization (GBEP-2) polysaccharides.The results were as follows:There is no significant difference of phagocytosis percentage,phagocytic index between lower dose group of GBEP-l,GBEP-2 and model group.But this significant differences reside in the larger dose group and model group (p<0.05或p<0.01) .The GBEP-1 and GBEP-2 all have augmented function to phagocytosis percentage and phagocytic index as well as correspond dose-effect.The results of hemolysin level are same to phagocytosis percentage and phagocytic index.The thymus index between lower dose group of GBEP-l,GBEP-2,larger dose group of GBEP-1 and model group have no significant difference.But thymus and spleen index differences reside in larger dose group of GBEP-2 and model group (p<0.05或p<0.01) .4.To purify the gingko biloba sarcotesta polysaccharide with DEAE-52 cellulose and sephadex G-100column chromatography.Obtain pure gingko biloba sarcotesta GBEPP11,GBEPP22,GBEPP33 and GBEPP44. The purify of them are 90.87%,95.36%,92.57% and 94.63% ,respectively.To determine the molecular and the compose of monosaccharides by using HPLC and GC.The result were as follows:The molecular of the four polysaccharides were GBEPP11: 950604, GBEPP22: 311288, GBEPP33: 47987, GBEPP44: 3373.The monosaccharides compose of GBEPP11 are rhamnose,glucose and galactose;molar ratio is rhamnose:glucose : galactose=16:10: 0.96. The monosaccharides compose of GBEPP22 are rhamnose,glucose and galactose;molar ratio is rhamnose:glucose : galactose=2.64:1:1.43. The monosaccharides compose of GBEPP33 are rhamnose,glucose and galactose;molar ratio is rhamnose:glucose : galactose=37.01:1:8.46. The monosaccharides compose of GBEPP44 are rhamnose and glucose;molar ratio is rhamnose:glucose =1.9:1.5.To study the anti-tummor in vitro experiment of crude,deproteinization, pure ( GBEPP1,GBEPP2,GBEPP3,GBEPP4) and acetylated GBEPP1 polysaccharides on AGS,SGC and U937 by using MTT method.The conclusion is:all of them have effect on AGS,SGC and U937,but the inhibition ratio are variously and the function of acetylated GBEPP1 is the best of them.