Preparation And The Structure-Activity-Relationship Study Of Sulfated Salvia Polysaccharide

Polysaccharide was a kind of natural biomacromolecule.And it has been payed close attention to,because of the wide physiological activities,especially the pharmacal activities. Salvia was the orthodox Chinese crude drug,which possess a considerable position in the traditional Chinese drug.There were many researches about salvia,but only a few of them were about polysaccharides of Salvia.Through previous researches of polysaccharides,the extraction condition,sulfated condition,structure determination,and the structure-activity relationship were studied.The main results were got as follows.(1)The extraction condition of polysaccharide of Salvia was studied.The crude polysaccharides were extracted with hot water.Three factors in extraction were tested,such as extraction time,liquide ration:solid,extraction frequency.By means of response orthogonal test and statistic software STATGRAPHICS plus 5.1,the optimization extraction condition was established.The extraction time was 106.37min,the liquide ration: solid was 148.68:1,and extraction frequency was thrice.(2)The two methods of deproteinization were compared.The freeze thawing method was better than chemical method,and had a lower expense of the product,Freeze thawing method was the best selection to deproteinate in polysaccharide.(3)A principal component named SP1 was obtained through gel chromatography method.(4)By means of response orthogonal test and statistic software STATGRAPHICS plus 5.1,the molecule modification of SP1 was studied and the influence of three factors to the sulphur content of product was made clear,and the mathematical model has been established as well.(5)Compared with the general methods,a more feasible new method to measure the sulphur content of Sulfated polysaccharide was established.(6)The structure of SP1 and sulfated SP1 were confirmed by chemical and instrumental analysis methods.The results indicated:the molecular weight of SP1 was 234kd;SP1 was composed of six monosaccharides that is rhamnose,arabinose,glucose,and a few of rhaglucuronic acid,xylose and galactose,the proportion of the three main monosaccharides were 2:3:18;SP1 has three types of polysaccharide bond that is 1,4(2)-,1,3-,and 1,6-(or non-deoxidized terminal),and the proportion is 2:12:3;the molecular weight o f sulfated SP1 was heavier than SP1,and the variety of monosaccharides was same as SP1,but the content of glucose was decreased;the proportion of 1,4(2)- was also decreased.And the spatial structures of the both were not triple helix.(7)Both of SP1 and SPS can quench the fluorescence of BSA,and each of them was a single static quenching process.BSA was strongly binded to SP1 with the molar ratio of 1.45:1,and the ratio wasl.47:1 when binded to SPS.The binding distance between SP1 and BSA was 3.65nm,and between SP1 and BSA was 3.33nm.The hydrophobic force was the main binding force.The samples cannot make the conformational changes of BSA,studied by synchronization fluorescence.(8)With the cyto-active experiment,the activity of SP1 and sulfated SP1 were studied. The structure activity relationship of SP1 and sulfated SP1 were confirmed.The results show that the anti-HIV activities of products were related to their sulphur content,the activityof sulfated SP1 was higher than SP1.The sulfated SP1 reached higher activity when their degrees of sulfonation were suitable.And the sulfated SP1 which had a higher ratio of 1,2(4)-glucosidic bond reached higher activity.