Studies On Extraction, Purification And Preliminary Antitumor Activity Of Polysaccharide From Galla Chinensis

ObjectiveGalla chinensis has been used for various purpose in healthcare and disease treatments, but researches on polysaccharide from Galla chinensis(GCP) are much rarer. The main purpose of the dissertation was to study the establishment of extraction, separation, purification of GCP, structural determination and preliminary antitumor activity. The following studies can provide references for application and exploration of Galla chinensis resources and theoretics gist for exploration of functional product.Methods(1) The orthogonal assay was employed to test the effects of four factors including folds of water, extracting temperature, extracting time and extracting power on the yield of polysaccharides from Galla chinensis. The optimal sedimentation techniques of polysaccharide were determined by concentrating rate and amount of precipitator.(2) To select the best deproteinization method by using Bradford method with deproteinization rate and polysaccharide-remaining rate as indicators.(3) The GCP was separated as GCP-1、GCP-2and GCP-3with DEAE-52cellulose column, Sephadex G-75column.(4) The three polysaccharides were tested the relative molecular weight by HPGPC.(5) The three polysaccharides were tested combined monosaccharide by HPLC with PMP pre-column derivatization.(6) The in vitro antitumor activities of three samples against Hepg2cell were studied by MTT method.Results(1) The optimized parameters of extracting polysaccharides by ultrasonic were as follows:ultrasonic power700W, material/solvent ratio1:20, extracting30min at60℃. The optimal sedimentation techniques of polysaccharide were as follows:using95%ethanol as precipitator, concentrating the extract liquid to2.5times and adding3times of precipitator.(2) The optimal techniques of deproteinization from polysaccharide were using the method of enzyme and Sevag together.(3) According to the results of HPGPC, the molecule weights of GCP-1> GCP-2and GCP-3were182,2.1×104and6×104, respectively. Based on this, GCP-1may be a kind of oligosaccharide.(4) The test of monosaccharide components was carried out by HPLC with PMP pre-column derivatization. The results showed that GCP-2and GCP-3were mainly combined with Gal.(5) The pharmacology experiment indicate that the three samples have little inhibitory effects on the proliferation of Hepg2cells.Conclusion(1) Based on data from the single factor test, the optimum process of ultrasonic extraction were obtained. The optimal sedimentation techniques of polysaccharide were determined too.(2) The GCP was separated as GCP-1. GCP-2and GCP-3with DEAE-52cellulose column, Sephadex G-75column. The test results showed the main component and relative molecular weight of three polysaccharides. The identification results showed that the samples were pure without including protein and nucleic acid by UV scan. We also can get the preliminary structure information by IR scan.(3) The GCPs showed no obvious antitumor activity. Some other pharmacology experiments will be needed for searching its immune activities.