Study On Extraction,Isolation,and Antioxidant Activities Of Active Compositions From Ganoderma Lucidum

Ganoderma lucidum,as a kind of medicinal fungus,has more than two thousand-years history and the extremely important pharmacological activity.The research proves that the activity of G.lucidum polysaccharide and triterpene substances is the best materia.Its can strengthen immunity,anti-tumor,antioxidant,fall blood sugar,blood lipid and other effect.Its can widely used in health care,food,cosmetics,and other industries.G.lucidum cultivated with substitute materials in guanxian county that is the largest planting base in China.In this experiment,G.lucidum was from guanxian county.Aiming to provide theoretical basis for rational development of G.lucidum in guanxian county by studying the active components of G.lucidum.Firstly,the best extraction method is selected by five different extraction methods,it include that hot water extraction,ultrasonic crushing extraction,micro-blog assisted extraction,ultrasonic cleaning extraction and enzymatic assisted extraction.Then the extraction process was optimized by hot water extraction method and ultrasonic crushing method.On the basis of single factor orthogonal optimization and response surface optimization tests were conducted.Secondly,the purification,structure characterization,and antioxidant activity of G.lucidum polysaccharides were studied.Finally,the alcohol extract of G.lucidum was extracted,purified,and its antioxidant activity was studied.The main conclusions were as follows:By comparing the yield of G.lucidum polysaccharides,the five extraction methods of polysaccharides include that hot water extraction,ultrasound-assisted extraction,ultrasound assisted extraction,microwave-assisted extraction and enzyme-assisted extraction.According to the extraction yeild,the extraction order was M-GLP>E-GLP>U₂-GLP>H-GLP>U₁-GLP.On the basis of single factor and orthogonal process optimization and the response surface optimization test of hot water extraction method,studies have shown that the best combination was:liquid ratio is 10:1 mL/g,time is 2 h,temperature is 90?,Response surface results showed that the optimal combination was:liquid-material ratio was 17.71:1mL/g,time was 2.56 h,extraction temperature was 89.20?.Base on the single factor,The response surface optimization test was used for ultrasonic crushing method.The results showed that the optimal extraction process of polysaccharide from G.lucidum by ultrasonic crushing method as follow:liquid to solid ratio is 25:1 mL/g,time is 60 min,extraction power 760 W,and the extraction yield of polysaccharide from G.lucidum was 3.60%.Using activated carbon,chitosan and H₂O₂ method to optimize the depigmentation process of G.lucidum polysaccharide,on the basis of single factor experiment,the study showed that H₂O₂ was superior to activated carbon and chitosan.The HCl method and Sevage method were used to the deproteinization experiment of G.lucidum polysaccharides.The results showed that the Sevage method was superior to the HCl method,and the Sevage method was optimized of the orthogonal process.The results showed that the optimal combination is:shock time was 30 min,and the proportion of the sample solution to Sevage reagent was 4:1?V:V?,and the proportion of chloroform and n-butanol was 4:1?V:V?.The structure characterization of polysaccharides from G.lucidum extracted through five different methods and ethanol grading deproteinized G.lucidum.The results showed that galactose was dominant in ultrasonic crushing,while glucose was dominant in other polysaccharides.The polysaccharides from G.lucidum were similar in structure but the conten is differentt.GLP40,GLP60,and GLP80 are all ?-pyranose.The four polysaccharides have the same monosaccharide components,but their molar ratios are different.By analysis of antioxidant activity of G.lucidum polysaccharide?M-GLP,E-GLP,U₂-GLP,H-GLP,U₁-GLP?,ethanol fractional precipitation to protein removal G.lucidum polysaccharide?GLP,GLP40,GLP60,GLP80?,ethanol fractional precipitation ultrasonic broken G.lucidum polysaccharide?GLP20,GLP30,GLP40,GLP50,GLP60,GLP80?,G.lucidum polysaccharide pigment was removal?chitosan,activated carbon,H₂O₂?,G.lucidum polysaccharide protein was removal by different method?GLP-S,GLP-HCl?have antioxidant activity,There was a positive correlation between activity and concentration.Study of the optimization orthogonal with G.lucidum alcohol extraction process and antioxidant activity,base on the extract yield,the results show that the best combination was follow:liquid-to-solid ratio is 40:1?mL/g?,and the volume fraction is 40%ethanol,temperature is 50?,time 2.5 h,using petroleum ether,ethyl acetate,N-butanol,extraction with ethanol in alcohol extraction,by the study showed that the DPPH radical scavenging ability,reducing power,hydroxyl radical scavenging capacity,the order as follows:V_C>N-butanol phase>Ethanol phase>Ethyl acetate phase>Petroleum ether phase,It antioxidant activity is relatively high close to the positive control that N-butanol phase and Ethanol phase.