Studies On The Chemical Components Of Ganoderma Lipsiense (Batsch) G.F.Atk. And Antitumor Activities In Vitro

In order to clarify the chemical and effective compositions of this medicine and their action, this research studied the chemical compositions and anti-tumor activities of G lipsiense.Solvent extracting method was employed to extract small molecular substances from G lipsiense.Roast G. lipsiense with oven at 40℃ and communite them into powder .The powder was stir-soaked with petroleum ether in stainless steel container at room temperature for 24 hours for several times.The superant was collected and combined, and distilled at 40℃ at a reduced pressure. Petroleum ether layer of extract liquid was obtained.The residual powder of G lipsiense, in which petroleum ether has been all volatilized, was soaked and extracted with 95% ethanol at room temperature.The ditto method was employed in this process. Ethanol layer of extract was obtained and extracted with diethyl ether, chloroform, ethyl acetate and n-butanol in turn, then extracts of the solvent above were obtained.Systematic solvent separation, two phases solvent extraction, chromatography, crystallization and re-crystallization methods were applied to separate and purify the chemical compositions in ethanol, petroleum ether and chloroform layers respectively and gradually by different isolation process.One water soluable crystal, crystal Ⅰ and four fat soluable crystals, crystal Ⅱ,Ⅲ, Ⅳ and Ⅴ was obtained after systematic isolation besides one volatile part, petroleum ether phase of part I in chloroform layer.Crystal Ⅰ was further isolated and purified by HPLC, then three compounds, compound L, M and N were obtained. Crystal Ⅱ was further isolated and purified by fractional crystallization and recrystallization, six crystals A-F were obtained.The configurations of crystals were identified on the basis of NMR, MS, HPLC, TC and thecomparison with normal collection of illustrative plates.The analysis results indicated that compound L was mannitol, compound N was (+ )-trehalose, and compound F was Ergosta-7, 22-dien-3p-ol.The chemical components of crystal Ⅲ, D, E and petroleum ether phase of part I in chloroform layer are analysised by GC-MS.The result indicated that Ergosta-14 , 22-dien-3-ol(3.beta., 5.alpha., 22E) and Cholest-4-ene, 3.beta.-(methoxymethoxy)-are the main components of crystal Ⅲ; Ergosterol and Ergosta-14, 22-dien-3-ol(3.beta., 5.alpha., 22E) are the main components of crystal D; 1, 2-Benzenedicarboxylic acid, diisooctyl ester, Eicosane, Tridecanol, 2-ethyl-2-methyl-, Hexadecane, 2, 6, 10, 14-tetramethyl- andErgosta-7, 22-dien-3-ol, (3.beta., 5.alpha., 22E)-are the main components of crystal E; 1-Dodecanol, 3, 7, 11-trimethyl-, Pentanedioic acid, dibutyl ester, Hexanedioic acid, bis(2-methylpropyl) ester , Pentadecanoic acid , 14-methyl- , methyl ester , 8 , 11-Octadecadienoic acid, methyl ester, 9-Octadecenoic acid, methyl ester and Linoleic acid ethyl ester are the main components of petroleum ether phase of part I in chloroform layer. All these components were found in G. lipsiense for the first time except Ergosterol and Ergosta-7, 22-dien-3-ol, (3.beta., 5.alpha., 22E)-.MTT is employed in this study to investigat the effect on MCF-7 cells' livability of the extracts of ethanol, diethyl ether, ethyl acetate, chloroform and petroleum ether layers.The results indicated that the extract of chloroform layer inhibited MCF-7 cells better and others did not manifested inhibition.The conclusion is the base of further pharmacological research.