Study On Polysaccharides Characteristics From Gymnadenia Conopsea R.Br.

This subject mainly in Yushu regions of Qinghai valuable Chinese herbal medicine Gymnadenia conopsea R.Br. as raw materials, Systematic go into the polysaccharide extracting technology, purification, structure analysis, and preliminary go into the toxicological characteristics for the development and to provide reference for the health care products.In this study, four extraction methods of GCP. conventional solvent extraction, microwave extraction method, ultrasound extraction method and enzyme extraction method were studied. Based on the single factor experiment, the optimum parameters and scopes were ensured by the method of orthogonal design. The result showed that the optimum parameters of solvent extraction were80℃,2.5h, and the ratio of material to water was1:30, extraction times were3times, the extraction rate of polysaccharide was up to11.83%; The optimum parameters of microwave extraction method were250W,20min, the ratio of material to water was1:30. extraction times was1time, the extraction rate of polysaccharide was up to13.56%; The optimum parameters of ultrasound extraction method were240w, ultrasonic extraction temperature60℃,10min and extraction times were3times, Under the optimized condition, the extraction rate of polysaccharide was15.24%; The optimum parameters of cellulose were pH4.8. temperature50℃.the adding of cellulose was1%of Gymnadenia conopsea R.Br..4h. the extraction rate of polysaccharide was16.58%.By treatment with alcohol-precipitation and by Sevage procedure, the small molecular (?)mpound and protein was excluded during isolation and purification. After concentrated and vacuum freeze drying, the GCP was obtained. This polysaccharide contains72.38%of sugar without free protein and small molecular compound, on this basis, the acute toxicity of the GCP was evaluated.GCP was further isolated into GCP-1and GCP-2a through the DEAE-cellulose column and sephadex G200gel chromatogram column. And then GCP-2a was collected and used to structure analysis. Identified by the infrared spectrum analysis, purification of GCP-2a with the characteristics of polysaccharide structure and functional groups:the sugar pyran ring carbonyl. hydroxyl and amino groups, etc. Phenol-Sulfating agent analysis showed that it contains polysaccharide92.6%;the Binding Proteins amount to0.87%according to Coomassie Brilliant; by vapor phase chromatography. GCP-2a is composed by Arabinose, xylose, mannitose, galactose and glucose.Gymnadenia conopsea R.Br. polysaccharides have no significant toxic effects and side-effects...