Studiey On Chemical Components And Functional Activities Of Polysaccharides From Malus Micromalus Makino Wine

Malus micromalus Makino is a perennial deciduous tree,which belongs to the genus Malu(Rosaceae family).It is a fruit with rare scarce resource only in China.Malus fruits taste sweet and sour,and they are rich in nutrients.Malus wine was brewed using Malus fruit juice as a raw material.Some chemical components were produced from original juiuce and metabolites of yeast in fermentation,which are of unique physiological activity and have high nutritional and medicinal value.In order to study the main compounds and their activities of Malus fruit wine,the chemical components were isolated from the wine,purified and identified,and the functional properties of polysaccharides from Malus wine was studied.There are five experimental parts in this dissertation:analysis and detection of aromasubstances in the Malus wine;isolation and identification of liposoluble components;isolation and purification of water-solublepolysaccharidesandstructuralcharacterization;evaluation of antioxidant and anti-aging activies of polysaccharide;and hypoglycemic activities of polysaccharides.The main results drawn are as follows:(1)Analysis and Detection of Aroma Components of the Malus Fruit WineThe aroma components of the fruits wine were analyzed by upper space-solid phase microextraction-gas chromatography-mass spectro-metry.The relative percentage of each component was determined by peak-area normalization method.The results showed that the fruit wine contained 60 aroma components,mainly alcohol and esters.Meanwhile,aldehydes,ketones,organic acids,and olefins were also detected.(2)Isolation and Identication of Liposoluble Components of the Malus WineThe Malus fruit wine was concentrated and extracted by petroleum ether,chloroform,ethyl acetate and n-butanol successively.The obtained extracts were separated by various charomatography(Silica gel thin layer chromatography,silica gel column chromatography,Sephadex LH-20 dextan column gel chromatography)and recrystallized.By means of physical and chemical properties and spectr oscopy,13components were identified from the extract of ethyl acetate:(1)hyperoside,(2)rutin,(3)catechin,(4)epicate-chin,(5)ursolic acid,(6)β-sitosterol,(7)naringin dihydrochalcone,(8)caffeic acid,(9)oleano-lic acid,(10)chlorogenic acid,(11)gentisic acid,(12)gallic acid,(13)vanillic acid,respectively.(3)PurificationandPrimaryStructuralCharactrtizationof Water-soluble PolysaccharidesThe extract was purified by macro-porous resin,separated by DEAE-52 cellulose chromatography column,screened by running water dialysis,Sephadex LH-20 glucan gel ccharomatography,and finally the pure polysacharide(MWP-21)was obtained from the fruit wine by vacuum freeze-drying.The primary structure of olysaccharide MWP-21was characterized by high performance gel permeation,ion chromatograph,UV Spectroscopy,Infrared Spectroscopy,mrthylation analysis and Nuclear Magnetic Resonance analysis.The results showed that the relative molecular mass of MWP-21 was 1×10~5 Da and it was composed of galactose,glucose and mannose and fructose in the mole ratio of 2.2:4.9:1.0.MWP-21 chain was composed of five sugar residues:α-1,6-Glcp,α-1,4-Galp,β-1,4,6-Glcp,α-1,3,6-Galp andα-1,2,6-Manp.(4)Evaluation of Antioxidation and Anti-aging Activities of polysaccharideThe free radical scavenging effect of polysaccharide was evaluated by antioxidant experiment in vitro.The results showed that crude polysaccharide(MWP)had the effect of scavenging hydroxyl,·DPPH and superoxide anion,and presented a dose-effect relationship.The results of anti-aging experiments in vitro showed that MWP could significantly reduce the lipid peroxidation level of brain cells,increase the activity of antioxidant enzymes(SOD,CAT and GPx),and remarkably inhibit the apoptosis of cerebral cortex cells,which proved that MWP can inhibit the decline of the brain by improving the body’s antioxidant capacity,inhibiting oxidative stress and reducing the apoptosis neurons in the cerebral cortex.(5)Research of Hypoglycemic Activity of PolysaccharideThe hypoglycemic activity of polysaccharides was studied with the body weight,blood sugar and insulin content in serum of mice as indicators.Malondialdehyde(MDA)content and antioxidant enzymes(SOD,CAT and GPx)activity in mouse hepatocytes were used as indicators to study the protective effect of polysaccharide on liver.The results showed that the polysaccharide MWP could decrease the blood sugar concentration of streptozotocin-induced diabetic mice,signify-cantly inhibit the weight loss of diabetic mice and promote the serum production in insulin.At the same time,MWP can obviously reduce the level of malondialdehyde in liver of diabetic mice and increase the activity of antioxidant enzymes(SOD,CAT and GPx).Furthermore,MWP can effectively improve the symptoms of liver injury such as fibrosis of liver tissue,enlargement of intercellular space,absence of central vein,and pseudolobular hyperplasia.It has a hepatoprotective effect to some extent.