Preparation Of Acanthopanax Senticosus-Ganoderma Lucidum Bidirectional Fermentation Product And Analysis On Its Main Active Components By LC-MS

Objective:Two-way solid fermentation was carried out with Acanthopanax senticosus as the drug matrix and Ganoderma lucidum as the fermentation strain,and the best fermentation process of Acanthopanax senticosus-Ganoderma lucidum was established.Through the qualitative and quantitative analysis of the main chemical components of the bud and the drug matrix,the change trend of the chemical composition of the fungus and the formation mechanism of the material foundation during the fermentation process were further elucidated.Methods:Using biotransformation technology,the fermentation time,biomass and bud active ingredient content were used as indicators to evaluate the two-way fermentation conditions of Acanthopanax senticosus and Ganoderma lucidum by single factor investigation and response surface test.Ganoderma lucidum and Sci-finder platform were used.The chemical constituents of Acanthopanax senticosus were integrated into multi-stage mass spectrometry information,HPLC-MS-DS was established,and its search function was applied to the rapid identification and efficient preparation of chemical components.The medium-pressure rapid purification preparative chromatography and preparative high performance liquid chromatography were used.Preparation of monomeric triterpenoids;Combined with HPLC-Q-TOF-MS/MS and HPLC-ESI-QQQ-MS/MS techniques,qualitative and quantitative study of the chemical composition of fermented bacteria,and the method of liquid chromatography-mass detection system Investigate and investigate the structural changes and cleavage of metabolites before and after fermentation,and dynamically track and analyze the changes in the main chemical components and content of the fungus.Results:The best technique for the two-way solid fermentation of Acanthopanax senticosus-Ganoderma lucidum is to weigh the powder of Acanthopanax senticosus（over 10mesh sieve）,add 40% distilled water by mass,MnCl₂ 82 mmol/L,NaCl 78 mmol/L,CaCl₂149 mmol/L.After mixing evenly,it was placed in a triangular flask and autoclaved twice at 121℃ for 30 min each time.Ganoderma lucidum shake flask seed solution wet weight（g）:acanthopanax senticosus solid medium（g）=2:1 ratio,the appropriate bactericidal age of the second Ganoderma lucidum shake flask seed solution is connected to Acanthopanax senticosus culture On the substrate,inoculated into a constant temperature and humidity incubator at 28℃,38%RH protected from light,until the Ganoderma lucidum mycelium covered the entire triangular flask and formed a distinct bud structure is the end of fermentation;Using HPLC-Q-TOF-MS/MS technology identified twelve chemical components such as Colossolactone IV in Ganoderma lucidum buds produced by two-way solid fermentation.Twelve chemical components such as Acanthopanaxoside C were identified in the matrix of Acanthopanax senticosus before and after bidirectional solid fermentation.Ganoderic acid P with a purity of 94.16% and Ganoderic acid T with a purity of 96.69% were obtained by chromatographic separation,and its chemical structure was confirmed by NMR;the Acanthopanax senticosus-Ganoderma lucidum was obtained by HPLC-QQQ-MS/MS technique.The dynamic tracking analysis of the relative contents of twelve chemical components such as Silphioside F,and determined that when the fermentation time reached 30 days,the changes of the main chemical components in the fungus became stable,which confirmed the existence of the fermentation end point.Conclusion:The chemical base and quality of Ganoderma lucidum and Acanthopanax senticosus were modified by two-way solid fermentation technology,which provided a theoretical basis for improving the bioavailability of medicinal materials and increasing the active substances.The medicinal properties of Ganoderma lucidum and Acanthopanax were separately applied.The matrix was studied to clarify the attribution of chemical components in the mixture of fungi.Separation and purification of the monomer Ganoderma lucidum triterpenoids,providing new resources for the acquisition of rare ganoderic acid;HPLC-MS-DS establishment,HPLC-Q-TOF-MS/MS technology and HPLC-QQQ-MS/MS technology combined qualitative Quantitative application,comprehensively identify the chemical constituents of the fungus and explore its pyrolysis pathway,truly evaluate the intrinsic quality of the fungus,and provide technical support for the multi-index quality control of the two-way fermented medicinal materials and preparations.