Study On Chemical Compositions In Liquid Submerged Fermented Mycelium Of Ganoderma Lipsiense

Ganoderma lipsiense(Batsch) G.F.Atk.belongs to the fungal genus Ganoderma in taxonomy,which is valuable in medicinal use.Modern medical and pharmacological researches have proved that Ganoderma lipsiense was active in improving immunity,resisting tumor,antivirus,antibiosis,eliminating inflammation, and so on.Thus in recent years the researches on chemical compositions in Ganoderma lipsiense have gained more and more attention.Ganoderma lipsiense collected from different habitats influenced by different geographical and environmental as well as other conditions differ in chemical composition constitution and content,which led to difference in its drug property and added to the difficulty in its practical application,To solve this tough problem,liquid submerged fermentation of Ganoderma lipsiense have caused extensive interest presently.To add new content to the systematic studies of the chemical compositions in Ganoderma lipsiense and facilitate its progress,as well as establish foundation for its further pharmacological studies and clinical application,this thesis made further investigation and exploration on the chemical compositions in the liquid submerged fermented mycelium of Ganoderma lipsiense on the basis of previous studies.Specialized qualitative methods were adopted to study the chemical compositions in the liquid submerged fermented mycelium of Ganoderma lipsiense in this thesis and carbohydrate,steroid,terpenoid, essential oil,organic acid,coumarin as well as alkaloid were detected.The mensuration vanillin-glacial acetic acid-perchloric acid spectrophotometric method used for detecting terpenoid content was optimized,and the optimum conditions were obtained,which provided reliable and sensitive mensuration for detecting terpenoid content in Ganoderma lipsiense.The optimum conditions were:φ(vanillin-glacial acetic acid)=5%solution 0.2ml,perchloric acid 0.8ml,70℃aqueous bath reacting for 20min,cooling in iced bath,plus glacial acetic acid 5.0ml,detecting wave length 570nm.Terpenoid contents in mycelium and fruiting body of Ganoderma lipsiense was 10.40%and 23.02%respectively detected through the optimized method.The chemical compositions in fermented mycelium were extracted through digeratur by organic solvents.The extractive, centrifugated under low temperature and low ethanol conditions,was separated and purified by silica gel column chromatography and macroporous resin adsorption chromatography,and 4 monomer compounds were obtained through TLC preparation,crystallization,recrystallisation,and so on.They were identified as ergosterol,mannitol and erythritol through nuclear magnetic resonance spectrum,UV and UR spectral analysis and with the aid of previous literature,among which erythritol was reported for the first time in Ganoderma lipsiense.Compositions constitution of unsaponifiable matters in fermented mycelium of Ganoderma lipsiense were firstly analyzed.Methanol-KOH saponifying method was employed to saponify the aether extractive from the mycelium and thus the unsaponifiable matters were obtained.Applying GC-MS and spectra retrieval,21 foregone compounds and 1 unknown compound were identified and analyzed,which were mainly divided into four categories:diolefine,acetylphenanthrene,sterol and butylated hydroxytoluene, Among them butylated hydroxytoluene,acetylphenanthrene,2-Bromo dodecane,1-iodo-Octadecane and 5, 6-Dinydroergosterol were all firstly reported in the chemical composition researches on Ganoderma lipsiense. The physico-chemical properties and structure of the unknown compound is being analyzed now.In the unsaponifiable matters the content of steroid and acetylphenanthrene were fairly high.Studies showed that steroid and acetylphenanthrene possessed many biological activities respectively,whether or not they would influence the pharmacological activity of the unsaponifiable matters because of their simultaneous existence and high content still need further pharmacological experiments to certify,so as to provide theoretical basis for correctly evaluating the medicinal value and establish foundation for further exploitation and utilization of liquid submerged fermented mycelium of Ganoderma lipsiense.