Extraction,Isolation,purification And In Vitro Immune Activity Of Polysaccharides From Cistanche Tubula

Objective: To establish an optimal extraction process for polysaccharides from Cistanche tubulosa dregs,and to investigate the effect of macroporous adsorption resins with different polarities on the decolorization and deproteinization of polysaccharides in aqueous extracts of Cistanche tubulosa,and to screen the best macroporous adsorption resins.Study its decolorization process.DEAE-650 M filler was used to separate the polysaccharide components in the water extract of Cistanche tubulosa and compare the antioxidant activity and in vitro immune activity of each component.Method: On the basis of single factor experiment,the polysaccharide extraction process was optimized by orthogonal experiment;the resin with the best decolorization effect was screened out by static adsorption method,combined with dynamic elution method,the main factors affecting the separation effect were studied,and the best Best separation process.According to the difference of the groups carried by each polysaccharide molecule,DEAE-650 M filler was used to separate neutral polysaccharides and acidic polysaccharides,and their ability to scavenge DPPH free radicals and hydroxyl free radicals in vitro,and to conduct preliminary research on the reducing ability of copper ions.Ascorbic acid(Vc)was used as a control;the effects of various polysaccharides on the secretion of nitric oxide,tumor necrosis factor and interleukin-6 levels of macrophages RAW264.7 were studied.Results: The optimal conditions for extracting polysaccharides from Cistanche tubulosa dregs were as follows: extraction temperature was 90℃,extraction time was 2h,solid-liquid ratio was 1:50;the extraction rate of polysaccharide under these conditions was 5.68%.Screening found that AB-8 macroporous adsorption resin has a better decolorization and deproteinization effect on the water extract of Cistanche tubulosa.The best decolorization process: loading concentration 25mg/ml,adsorption time 12 h,optimal flow rate 1.2BV/h,The purity of polysaccharides in the water extract of Cistanche tubulosa changed from 23.9% to 55.4%.Antioxidant experiments have found that the crude polysaccharide of Cistanche tubulosa with a certain concentration range has a better scavenging effect on DPPH free radicals and hydroxyl free radicals,and has a better reducing ability on copper ions.In the in vitro immune activity research experiment,it is found that each polysaccharide has a certain ability to regulate immune activity,among which acidic polysaccharides CT2 has strong anti-inflammatory ability.Conclusion: The extraction method is simple,accurate,energy-saving and environmentally friendly.It is suitable for the extraction of Cistanche tubulosa polysaccharides.AB-8 macroporous adsorption resin has a good separation effect on the polysaccharides in the water extract of Cistanche tubulosa.Operate and safe.DEAE-650 M ion-exchange column chromatography is also suitable for the separation and purification of Cistanche tubulosa polysaccharide.The purified Cistanche tubulosa has strong anti-inflammatory activity and good antioxidant activity.,It is convenient to popularize and use in the preparation of Xinjiang Cistanche tubulosa polysaccharides and lay the foundation for the development of Cistanche tubulosa products.