Study On Extraction, Separation And Purification Of Polysaccharide From Seabuckthorn

The main research object is to research several methods of extracting seabuckthorn polysaccharide. Using the traditional hot water extraction, microwave-assisted extraction method,ultrasound-assisted extraction method, respectively orthogonal test, is to fing the optimum extraction. The extracting of polysaccharides from seabuckthorn was precipitated by ethanol. Polysaccharide is purified by n-butanol■20%trichloroacetic, n-butanol-20%trichloroacetic method to remove protein, and then followed by DEAE-52column chromatography. Polysaccharide is shown to be a single homogeneous component by polyacrylamide gel electrophoresis and SephadexG-100column chromatography. The results showed that:1.Traditional hot water extraction of polysaccharide optimum conditions are as follows:The extraction temperature is90℃, extraction time is2h, water:angustifolia is30:1.The extraction content is13.96mg/g.Microwave-assisted extraction of Angustifolia polysaccharide optimum conditions are as follows:The extracting power is100W, extraction time is5min, water:seabuckthorn is30:1, The extraction content is11.96mg/g.Ultrasonic extraction auxiliary seabuckthorn polysaccharide optimum conditions are as follows:The Ultrasonic power is110W, extraction time is45min, extraction temperature is50℃, water:seabuckthorn is30:1, The extraction content is14.36mg/g. The optimal technique conditions for the enzymatic extraction were:the extraction time is45min, the extraction temperature is50℃, PH is4.5, and the Enzyme dosage is2%, under these conditions, the highest yield of polysaccharide from Hippophae fhamnoides L. is11.26mg/g. Considering various factors, Ultrasonic extraction is the optimal extraction method.2. The extracting of polysaccharides from seabuckthorn were precipitated by ethanol. Polysaccharide is purified by n-butanol,20%trichloroacetic, n-butanol-20%trichloroacetic method to remove protein, and compared with that three methods. The results showed that:it is the best way to remove protein from Polysaccharide using n-butanol-20%trichloroacetic method, the polysaccharides lose rate is33.23%and the protein removal rate is73.53%.3. The polysaccharides that has been removed protein followed by DEAE-52column chromatography, and we obtained two kinds of polysaccharides. And they are shown to be two single homogeneous components by PAGE and SephadexG-100column chromatography.