Study The Extracting And Purifying Technology And Immune Activity Of Polysaccharides From Chelidonium Majus L

Objective:Study the crafts of polysaccharide extraction and purification and immunological activity from Chelidonium majus L.Provide theoretical basis for further development and utilization of polysaccharides Chelidonium majus L.Methods:(1)Using glucose as standard products to establish a standard curve and use of sulfuric acid-phenol method,for the determination of polysaccharide.(2)Set polysaccharide yield and polysaccharide content as indices and use weighted score(weighting each 50%),inspect the single factor of solid-liquid ratio, extraction time, extraction temperature, extraction times, alcohol precipitation concentration respectively and optimization the best water extraction process by orthogonal test based on the best single factor.(3)Set polysaccharide yield and polysaccharide content as indices and use weighted score(weighting each 50%),inspect the single factor of alkali concentration, extraction time, extraction temperature, extraction times respectively and optimization the best alkali extraction process by orthogonal test based on the best water extraction process and alkali.(4)Set polysaccharide yield and polysaccharide content as indices and use weighted score(weighting each 50%),inspect the single factor of enzyme dosage, reaction pH, hydrolysis time, reaction temperature respectively and optimization the best alkali extraction process by orthogonal test based on the best enzyme.(5)Set decolorization and polysaccharide retention as indices and use weighted score(weighting each 50%) to determined the best decolorization process of activated carbon and hydrogen peroxide by orthogonal test based on relevant literature.(6)Set polysaccharide retention rate,bleaching and desorption rate as indices and use weighted score(weighting each40%、30%、30%) to determined the bestmacroporous resin of 8 types.Set decolorization and polysaccharide retention as indices and use weighted score(weighting each 50%) to determined the best purification process by the single fators of sample concentration,higher than the diameter, eluent flow rate, sample volume, the amount of eluent.(7)Take immunocompromised mice by intraperitoneal injection of cyclophosphamide,set saline for the control group,lentinan as; positive control group, the purified polysaccharide from Chelidonium majus L different doses of the experimental group,fed with an equal volume of the solutionto determined the immunocompetent and the best dose of polysaccharide from Chelidonium majus L by peritoneal macrophage phagocytic rate and phagocytic index and the formation of hemolysin and hemolytic plaque situation.Results:(1)The best water extraction process of polysaccharide from Chelidonium majus L:Take 30 times the amount of water to powder of Chelidonium majus Z,,100℃ extracted three times,each four hours. Optimal concentration of alcohol precipitation is 80%.(2)The best alkali extraction process of polysaccharide from Chelidonium majus Z,:Take 20 times the amount of 0.1 mol/L Na2CO3 solution to dregs of Chelidonium majus L,4℃ extract 1, every four hours.(3) The best enzyme extraction process of polysaccharide from Chelidonium majus L:Chelidonium majus L powder was added to 30 times the amount of water, soak for 40℃ 30 minutes, adding 5% of pectinase enzyme 2 hours under conditions of pH was adjusted to 3.0,40℃.(4)The best decolorization process of activated carbon:The polysaccharide from Chelidonium majus L solution was added 0.5% of activated charcoal, adjusted to pH 3.0,50℃ under the conditions of a water bath with stirring 40min; The best decolorization process of a hydrogen peroxide:The polysaccharide from Chelidonium majus L solution was added 10% hydrogen peroxide solution, under conditions of pH was adjusted to 10.0,50℃ water bath for 1h.(5)The best purification process of macroporous resin:Take HPD-600 resin to the ratio of diameter and higher is 1:8 column packed with the polysaccharide sample solution 0.04g/ml samples on 1BV,adsorption was allowed to stand 4 h, according to a flow rate of 1.0ml/min eluted with distilled water, the amount of eluant to 8BV.(6)The refined polysaccharides from Chelidonium majus L can significantly improve the immunocompromised mouse peritoneal macrophage phagocytosis of chicken erythrocytes percentage and phagocytic index, promote hemolysin and hemolytic plaque formation, and 0.05g/kg celandine polysaccharide group work best.Conclusion:Through this project obtained celandine extraction and purification process is stable and reliable,and the enzyme extraction process and macroporous resin purification process works best,polysaccharide yield was 16.38%, polysaccharide content was 70.17%.Pharmacological experiments also proved celandine polysaccharides can significantly improve the immunocompromised mice immune activity, are worthy of further study.