Separation And Purification Of Polysaccharide From Abelmoschus Manihot And Development Of The Solid Beverage

Abelmoschus manihot is a medicinal and edible plant,which contains flavonoids,polysaccharides,vitamin E and other active substances.Polysaccharide is the second highest active substance of Abelmoschus manihot,which has many effects such as anti-oxidation,anti-aging and anti-cancer.In this study,the polysaccharide of Abelmoschus manihot was extracted,separated and purified,and the physicochemical properties,structure and biological activity of Abelmoschus manihot polysaccharide were studied.The main results are as follows:1.Extraction and purification of polysaccharide from Abelmoschus manihot.The polysaccharide of Abelmoschus manihot was extracted by compound enzyme extraction method.The optimum extraction conditions of polysaccharide from Abelmoschus manihot were determined by single factor and orthogonal test: Six kinds of enzymes were used in this study,which were xylanase,pectinase,hemicellulase,papain,cellulase and glucose oxidase.The enzyme hydrolysis time was 40 min,and the ratio of material to liquid was 1﹕30.Under these conditions,the yield of crude polysaccharides from flowers,stems and leaves were 35.67%,22.67% and 26.67%,respectively.Then AB-8macroporous resin was used to purify the crude polysaccharide of Abelmoschus manihot.The decolorization rate of polysaccharide was used as the purification index of polysaccharide.The optimum purification conditions were optimized by single factor and response surface: resin addition amount 7g,shaking speed 180 r/min,shaking time 2.4 h.At this time,the decolorization rate of flower polysaccharide was 39.96%,the decolorization rate of stem polysaccharide was 44.23%,and the decolorization rate of leaf polysaccharide was 59.02%.2.Structure and composition analysis of polysaccharide from Abelmoschus manihot.The composition structure,glycosidic bond type and microstructure of polysaccharide were analyzed by ultraviolet absorption spectroscopy,Fourier transform infrared spectroscopy,scanning electron microscopy.The results showed that the polysaccharide had no obvious absorption peak at 260 ～ 280 nm and the polysaccharide contained α-type glycosidic bond and β-type glycosidic bond.The total sugar content of flower polysaccharides was 59.18%,and the reducing sugar content was 18.33%.The total sugar content of stem polysaccharides was 27.24%,and the reducing sugar content was 8.29%.The total sugar content of leaf polysaccharide was23.88%,and the reducing sugar content was 8.23%.3.Analysis of antioxidant activity of polysaccharide in vitro.Hydroxyl radical,superoxide anion radical,FRAP and reducing power were used as evaluation indexes.When the concentration was 1.0 mg/m L,the scavenging rates of hydroxyl radicals of the flowers,stems and leaves were 75.95%,26.16% and 28.90%,respectively.The scavenging rate of superoxide anion radical of flowers,stems and leaves were 87.26%,89.35% and 90.40%,respectively.The FRAP values of flowers,stems and leaves were 0.454,0.382 and 0.218,respectively.The reducing power of flowers,stems and leaves were0.098,0.059 and 0.107,respectively.4.The effect of Abelmoschus manihot polysaccharides on pancreatic cancer cells and heat-damaged spermatogonia cells.Abelmoschus manihot polysaccharide can promote the apoptosis of pancreatic cancer cells,and the order of influence is : medium dose > high dose > low dose,the apoptosis rate of pancreatic cancer cells was 14.06% at medium dose.At 18 h,the medium dose of flower polysaccharide had a certain repair effect on the spermatogonia cells treated with heat injury.At this time,the cell proliferation rate was41.04%,and the medium dose of the cells increased by 14.19% compared with the control.5.Development of Abelmoschus manihot solid beverage.Through single factor and orthogonal experiments,the optimum formula of solid beverage of Abelmoschus manihot was determined: the addition amount of Abelmoschus manihot,Codonopsis pilosula and Polygonatum odoratum was 0.5 g;the amount of sugar was 2.0 g;the addition amount of citric acid was 0.01 g;the addition amount of rice flour was 0.15 g;the addition amount of rice flour was0.2 g;the addition amount of millet flour was 0.1 g;the addition amount of yam powder was 0.15 g;the addition amount of corn flour was 0.20 g;the addition amount of blueberry powder was 0.2 g;the addition amount of cherry powder was 0.3 g;the addition amount of jujube kernel was 0.1 g;the addition amount of purple sweet potato powder was 0.3 g.Each 100 g of Abelmoschus manihot solid beverage contains 3.44 g of water and 11.85 g of reducing sugar.