Studies On Chemical Constituents And Quality Control Methods Of Ganoderma Lucidum

This dissertation includes four chapters:Chapter 1 Summary of the related literaturesThe regulation varieties of Chinese pharmacopoeia were the dry fruiting bodies of Ganoderma lucidum(Ley-ss.ex Fr.)Karst.and Ganoderma sinense Zhao,XuetZhang.In this part,the related literatures about the chemical constituents and pharmacological activities of Ganoderma lucidum were reviewed on the basis of 155 references,and summarized the constituents of triterpenes,polysaccharides,steroid body,fatty acids,alkaloids in the Ganoderma lucidum.It also has a wide range of pharmacological activities,such as anti-tumor,anti-inflammatory,anti-aging,regulation of immune function,lowering blood sugar and blood fat.The reaserches on quality control mostly focused on dertermination and fringerprints of active constituents.Chapter 2 Studies on chemical constituents from fruiting bodies of Ganoderma lucidumSolvent extraction method and several modern chromatographic techniques(including silica gel,C18,Sephadex LH-20,MCI,etc.)were used to isolate the chemical constituents of ethanol extract of the fruiting bodies of Ganoderma lucidum,and the structure of chemical constituents were identified according to physicochemical properties and spectralanalysis(ESI-MS,13C NMR,1H NMR,and ROESY,HMBC,HSQC,2D-NMR,etc.).Fourteen compounds were isolated from the fruiting bodies of Ganoderma lucidum,including twelve triterpenes and two cerebrosides.Twelve triterpenes were characterized as 3β,7β,15a-trihydroxy-11,23-dioxo-lanost-8,16dien-26-oic acid,3β,7β,15β-trihydroxy-11,23-dioxo-lanost-8,16-dien-26-oic acid methyl ester,3β,7a-dihydroxylanosta-8,24-dien-11-one,3β,7β,15a,28-tetrahydroxy-11,23-dioxo-lanost8,16-dien-26-oic acid,Ganotropic acid,methyl ganoderate C2,Ganosineniol A,Ganoderic acid B,Ganoderiol H,Ganoderic acid A,Ganoderic acid C,Ganoderic acid B.Two cerebrosides were characterized as(4E,8E)-N-D-2’-hydroxypalmitoyl-1-O-β-Dglucopyranosyl-9-methyl-4,8-spingodienine,Termitomycesphins G.Among them two triterpenes were first reported from the fruiting bodies of Ganoderma lucidum;one cerebrosides were first isolated from Ganoderma.Chapter 3 Studies on quality control method for Ganoderma lucidumThe HPLC fingerprints of fat-soluble components of Ganoderma lucidum from 15 different habits were constracted with the detection wavelength 254nm and analyzed by using Chromatographic Fingerprint Evaluation System(2004 A version).There were 37 common peaks in the fingerprints.The similarity analysis of 15 samples from different regions revealed that the similarities were between 0.956 and 0.772 with 4 regions above 0.9,and 9 regions above 0.8.Clustering results showed that 15 batches of G.lucidum were clustered into four categories.In the principal component analysis,six principal components were extracted,and the accumulated contribution rate was 93.69%,in which the first principal component contribution rate was 43.09%.The peaks 10,3,30,9,4 contributed the most to the first principal component.The content of the 5 components may be the main reason for the clustering result.The qualitative identification of the common peaks of the fat-soluble components of G.lucidum fingerprints were carried out by using LCMS-IT-TOF.33compounds in the 37 common peaks were identified.Common peaks,in some extent,are reflections of fingerprint informations and chemical constituents of medicinal materials.So it can provide a reliable basis for the quality control of G.lucidum from different regions by qualitative analysis of common peaks.A quantitative method was firstly established for the determination of Ganoderic acid A,B,G and 3β,7β,15α-trihydroxy-11,23-dioxo-lanost-8,16-dien-26-oic acid(GS8)by using High Performance Liquid Chromatography.The regression equations of thefour compounds were,Y=972162X-51942(r=0.9992),Y=836496 X-2621.6(r=0.9991),Y=834617X-3224.8(r=0.9992),Y=688859X+6086.6(r=0.9993),respectively.The linear range of the four ganoderic acids were,0.1575～2.6563 ug、0.0891～1.485 ug、0.1041～1.735 ug、0.1509～2.515 ug.The average recoveries were 101.68%with RSD of 2.10%for Ganoderic acid A,105.10%with RSD of 2.49%for Ganoderic acid B,100.24%with RSD of 3.72%for Ganoderic acid G,99.2%with RSD of 2.89%for 3β,7β,15α-trihydroxy-11,23-dioxo-lanost8,16-dien-26-oic acid,respectively.The content determination results of the four compounds in 15 batches of raw drugs showed that the methods were simple,quick,reliable,reproducible and can be used as the determination of G.lucidum.Chapter 4 Conclusion and discussion1.Fourteen compounds were isolated from the fruiting bodies of Ganoderma lucidum,including twelve triterpenes and two cerebrosides.Among them two triterpenes were first reported from the fruiting bodies of Ganoderma lucidum;one cerebrosides were first isolated from Ganoderma.2.The HPLC fingerprints of fat-soluble components of Ganoderma lucidum from 15 different habits were constracted and analysed by by using Chromatographic Fingerprint Evaluation System.The qualitative identification of the common peaks of the fat-soluble components of G.lucidum fingerprints were carried out by using LCMS-IT-TOF.33 compounds in the 37 common peaks were identified.3.A quantitative method for the determination of Ganoderic acid A,B,G and 3β,7β,15α-trihydroxy-11,23-dioxo-lanost-8,16-dien-26-oic acid(GS8)were firstly established by using HPLC.4.This topic used the fruiting bodies of Ganoderma lucidum as the research object,and for the chemical constituents and quality control method research,is mainly to triterpenoid compounds indicators,but pharmacologically active ingredients in Ganoderma lucidum is not only triterpenoid compounds,including polysaccharides and other compounds.In order to achieve the comprehensive evaluation and understanding of Ganoderma lucidum,studies on other ingredients are essential.