Study On The Extraction, Purification And Biological Activity Of Polysaceharides From Rosa Multiflora Thunb

In the research, Polysaccharides of Rosa multiflora Thunb were choosed as material,systematieally studied that extraction, separation,Purifieation of poiysaccharides from Rosa Multiflora Thunb.We studied the antioxidant and bacteriostatic functions on the Polysaccharides from Rosa Multiflora Thunb,which established a basis for them in foodstuffs, medieine and health supplies, etc.The main results and innovations were as follows:(1)The effects of different extraction methods on the yields of polysaccharide extracted from Rosa Multiflora Thunb were investigated.Different extraction methods,including traditional hot water extraction, microwave assistant extraction, ultrasound extraction,ultrasonic-microwave assisted extraction,enzymatic extraction and ultrasonic-microwave-enzyme synergistic extraction, were used to extract polysaccharides from Rosa Multiflora Thunb(RMTP),and the ultrasonic-microwave assisted extraction and ultrasonic-microwave-enzyme synergistic extraction methods optimized by response surface analysis(RSA).The results showed that the optimum technology conditions for ultrasonic-microwave assisted extraction of polysaccharides from Rosa Multiflora Thunb were : extraction temperature 81?, microwave power317 W, extraction time 22 min, the ratio of liquid to material 32:1(mL/g). The yield of polysaccharides was 6.10mg/g;and the results showed that the optimum technology conditions for ultrasonic-microwave-enzyme synergistic extraction of polysaccharides from Rosa Multiflora Thunb were : the amount of enzyme added was 1.15 %(enzyme activity was 40 U/mg), pH value was 5.72, extraction temperature was 81.19 ? and the ratio to liquid was 12.56:1(m L/g), at this conditions, actual yield of Rosa Multiflora Thunb polysaccharides was 10.48 mg/g.(2)To screen the optimum technology for deproteinizing polysaccharides from Rosa Multiflora Thunb.The different methods of removing protein including Sevag,TCA(trichloroacetic acid),enzyme,TCA-enzyme combination were compared with the rate of deproteinization as key indexes, and the TCA method optimized by response surface analysis.The optimal conditions of TCA method was as follows : The mass fraction of TCA was 0.9%,volume ratio was 1:1,standing time was 70 min.At this conditions, the rate of deproteinization was 75.92% and polysaccharide loss was22.57%.(3)The study aimed to discuss the optimum technology condition for the hydrogen peroxide method decolorization of polysaccharides from Rosa MultifloraThunb.The polysaccharides were discolored using hydrogen peroxide method,and the best decolorization conditions were determined by response surface methology.The optimum technology conditions for hydrogen peroxide method decolorization of polysaccharides from Rosa Multiflora Thunb was as follows:The volume fraction of H2O2 was 35.0%,pH value was 9.0,decolorization time was 4.5 h.Under such conditions the decolorization rate and retention rate of polysaccharide were 95.48%and 75.17%,respectively.(4)The purification method of Rosa Multiflora Thunb polysaccharide was set up,and using the DEAE-52 cellulose to purify the polysaccharides from Rosa Multiflora Thunb,and the optimized its best purification process.Results,the best purification process parameters were:the ionic strength were:water,0.1mol/L and0.3mol/L NaCl solution,quantity of DEAE-52 cellulose 30 mg,eluent velocity30mL/h.Three components RMTP-1,RMTP-2 and RMTP-3 were gained by DEAE-52 cellulose, purity identification results showed that purified components was a uniform component by freezing-and-thwaing test and UV spectrum.The UV specturm's result indicated that this Polysaccharide component does not contain nucleic acid, protein and other impurities.(5)Characteristic reaetions of RMTP-1 and RMTP-2 were diseussed, the resultss howed that it was non-starch Polysaccharide without protein or polyphenol material,and the has the connectivity of Polysaccharide compounds.Infrared spectrum of polysaccharides RMTP-1 and RMTP-2 indicates that they had typical characteristic absorption peaks.Monosaeeharide composition analysis by GC-MS chromatography of RMTP-1 and RMTP-2 show that : RMTP-1 of mainly consists of xylos,fructose,mannose;RMTP-2 mainly consists of xylos,fructose.(6)Test Rosa Multiflora Thunb polysaccharides in vitro antioxidant activity in a chemical way, and compared with the antioxidant activity of VC, and the antibacterial activity was evaluated by the methods of disk diffusion and standard tube dilution.The results showed that the polysaccharides from Rosa Multiflora Thunb had strong antioxidant capacity.The polysaccharides from Rosa Multiflora Thunb had weaker antioxidant activity than VC.The antioxidant activity and polysaccharide content in certain concentration ranges were related.The polysaccharide from the Rosa Multiflora Thunb had strong inhibiting effects on some pathogenic bacterium such as Bacillus subtilis ? Escherichia coli ? Staphylococcus aureus and Proteus mirabilis.