| Based on a large number of resources investigation and literature systematization, the medical bioactive ingredients of Marchantia convoluta living in Guangxi China were made as object. During pharmacological research, the liposoluble constituents of the plant were analyzed, extracted, separated and purified, and the crude drug identification was engaged. Then bioactive ingredients - ethyl acetate extracts were obtained which possessed the action of inhibition of liver cancer cells HepG2 and the small cell lung cancer cells H1299. The qualitative analysis was carried out by means of HPLC, UV, IR, NMR, MS, and GC-MS to determine the structure of the monomers. Details are as following:1.Identification of the plant samples. Three species of Marchantiale plants in Guangxi {Marchantia polymorpha, Marchantia convoluta and Marchantia paleacea) were identified through Ultraviolet Absorption Spectra Lines Group (UASLG). The UV spectra of the solutions extracted by four different kinds of solvents (water, chloroform, ethanol and petrol). The UV spectra of the three herbal drugs gave different maximum absorption peaks both in number and intensities. The method was both simple and accurate and can be used to distinguish one from others.2.Analysis of liposoluble constituents with GC-MS. The gas chromatography-mass spectrum was set up to identify liposoluble constituent in Marchantia convoluta. There were many differences in chemical constituents of the petroleum ether extracts and the ethyl acetate extracts. The fatty acids and their esters were the main constituents of the petroleum ether extracts. (Z)-11-Hexadecenoic acid and n-Hexadecenoic acid were the main fatty acid accounting for 9.80 and 20.29 % respectively in the petroleum ether extracts. There were few terpenoids in the petroleum ether extracts. But terpenoids were the main constituents of the ethyl acetate extracts. Phytol, benzothianzole and cedrol were the main compounds accounting for 25.80, 14.97 and 20.69%respectively in the ethyl acetate extracts. The cedrol was first identified from Liverworts.3. Extraction of volatile oil. Five factors and four levers orthogonal test methods were used to optimize the SFE conditions. The best extraction conditions were established: pressure, 15KPa; temperature, 50°C; extracting time, lh; modifier, alcohol (20mL). The SFE method provides many advantages, such as shorter extraction time and high extraction efficiency.4. Separation and identification of monomer in the petroleum ether extracts. Two sterol compounds (6-sistosterin and stigmasterol) were yielded by separating the lip soluble part with silica-gel column chromatography. The structures were identified by means of physic-chemical method combined with the application of UV, IR, HNMR and MS.5. Bioactivity testing of ethyl acetate extracts. The ethyl acetate extracts could evidently inhibit the small cell lung cancer cells HI299 and the liver cancer cells HepG2. But the petroleum ether extracts have no effects on the small cell lung cancer cells HI299 and the liver cancer cells HepG2. The average inhibition rates of the ethyl acetate extracts (5 ng/ml, 15 ug/ml, 30 ug/ml, 40 ug/ml) on liver cancer cells HepG2 were 42.88 >. 53.66^ 57.31 and 69.17% respectively. The average inhibition rates of the ethyl acetate extracts (5 ug/ml, 15 ug/ml, 30 (xg/ml, 40 ug/ml) on the small cell lung cancer cells H1299 were 6.31 > 17.65 ^ 18.96 and 23.89 % respectively.There have been no similar domestic and international documents in this field, which has been proved by authority. |
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