| Ganoderma lucidum(Leyss.ex Fr.)Karst,a medicinal fungus,was included in the “Sheng Nong's herbal classic” and Chinese Pharmacopoeia(edition 2015).And G.lucidum.has been extensively used to treat immune disorders,high blood pressure,chronic hepatitis,nephritis,gastric ulcer and cancer.G.lucidum has many active substances,which were widely and deeply studied.However,the G.lucidum polysaccharides were complex and indissolvable,which hindered the research on its extraction,biological activity and product development.Therefore,the G.lucidum polysaccharides and its related products were researched in this paper to provide scientific basis for pharmacological research of GLP,and contributing to the development of products or drugs of G.lucidum.In this thesis,the chief research content is presented as follows: enzyme ultrasonic/ microwave assisted extraction conditions of GLP were optimized through the response surface methodology(RSM).Then,the purified components of polysaccharides were obtained by Sevag method,DEAE-52 anion exchange column chromatography and Sephadex G-100 dextran gel column and characterized by HPLC-RID,HPLC-DAD and FT-IR.Finally,the quality control of G.lucidum tea was conducted,the preparation processes and quality control method of G.lucidum tea were developed.The main results are shown as follows:(1)Study on the enzyme ultrasonic/ microwave assisted extraction of GLPThe optimum extraction conditions of GLP were obtained by suitable enzyme types,single-factor test,and the response surface methodology,basing on GLP extraction rate.The results found that the mix-enzyme(cellulase,hemicellulase and pectinase = 1: 1: 1)could significantly improve the extraction rate of GLP.And the optimal extraction conditions obtained were that enzymolysis temperature 38 ?,enzyme dosage 1000 U,solvent dosage 22.0 m L,ultrasonic time 49 min.Under this condition,GLP extraction rate was increased 106.9% by comparing with the traditional hot water extraction method.(2)Isolation,purification and structure identification of GLPGLP was isolated and purified by Sevag method,DEAE-52 ion-exchange cellulose and Sephadex G-100 dextran gel.And three fragments were obtained,named as GLP-D1-1,GLP-D2-1 and GLP-D2-1.Then,HPLC-RID,HPLC-DAD and FT-IR were used to characterize the purified components of polysaccharides.The results of HPLC-RI indicted that GLP-D1-1,GLP-D2-1 and GLP-D2-2 were homogeneous components,and the molecular weight(Mw)was 6.542 × 104 Da,2.007 × 105 Da,1.055 × 105 Da,respectively.HPLC-DAD analysis showed that GLP-D1-1,GLP-D2-1 and GLP-D2-2 were hetero-polysaccharides,the GLP-D1-1 was mainly composed of D-mannose,rhamnose,D-galacturonic acid,glucose,D-galactose,L-fucose in the molar ratio of 20.7: 2.13: 1.18: 45.57: 27.67: 2.75;the GLP-D2-1 was mainly composed of D-mannose,rhamnose,D-glucuronic acid,D-galacturonic acid,glucose,D-galactose,L-fucose in the molar ratio of 12.81 : 5.92: 1.74: 0.84: 68.22: 8.84: 1.63;the GLP-D2-2 was mainly composed of D-mannose,rhamnose,glucose,D-galactose in the molar ratio of 6.11: 5.59: 82.71: 5.59.FT-IR analysis showed that GLP-D1-1,GLP-D2-1 and GLP-D2-2 had typical polysaccharides characteristic bonds and specific spectral signals.The sugar configurations of GLP-D2-1 and GLP-D2-2 were speculated as ?-type.(3)Study on the production technology and quality standard of Ganoderma Lucidum teaIn order to optimize the best production process parameters of G.lucidum tea,the Box-behnken response surface methodology was used to optimize the production process based on the results of single factor experiments.The optimum conditions were as follows: the dosage of G.lucidum extract was 0.93 g,the baking time was 61 min,and the baking temperature was 79 °C.Under these conditions the maximum value of sensory evaluation score of G.lucidum tea was 86.1.In order to control the quality of G.lucidum tea,the methods of determining ganoderic acid A and ganoderic acid B were established by ultra high performance liquid chromatography electrospray ionization tandem mass spectrome.The results indicate that Ganoderic acid A and ganoderic acid B had excellent linearity within the concentration range of 0.0513 ~ 0.5130 ?g·m L-1,0.0208 ~ 0.1247 ?g·m L-1,respectively.The average recoveries were as follows: ganoderic acid A was 99.57%(RSD = 2.74%,n = 9),ganoderic acid B was 101.9%(RSD = 1.08%,n = 9).The methods of determining polyphenols and polysaccharides were established by using UV-spectrophotometry.The results indicate that Gallic acid and glucose had excellent linearity linearity within the concentration range of 4.08 ~ 16.50 ?g·m L-1,40.33 ~ 181.5 ?g·m L-1 respectively.under this method,the average recoveries were as follows: 99.6%(RSD = 2.52%,n = 9),100.5%(RSD = 1.08%,n = 9).The result showed this study provides some scientific basis and methods to control the quality of G.lucidum tea. |
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