Study On Isolation And Identification Of Phoebe Sheareri And Its Bioactivities

Phoebe sheareri is a genus of Phoebe,It's tall trees mostly.Phoebe and Machilus are collectively known as Nanmu in China.In the south of the Yangtze River in China,Phoebe sheareri is a associated species in the natural evergreen broad-leaved forest.Phoebe sheareri is a timber tree species,for the construction,precious furniture,It is a important economic forest in China.Phoebe sheareri play an important position in the garden,wood,spices and other aspects.Phoebe sheareri plant resources are rich in China.It can be used as herbs,with more significant biological activity,such as analgesic,anti-inflammatory,anti-bacterial and anti-infection.In the first section,the latest progress of extraction and purification of plant active ingredients is described.At the same time,the composition analysis and biological activities of Phoebe sheareri were studied systematically and meticulously.Nanmu mainly contains chitosan,flavonoids,butyrolactones,alkaloids and phenolic compounds,It has various pharmacological activities with liver protection activity,neuroprotective activity,cytotoxic activity,Caspase-3 enzyme activity and so on.At the end of the this chapter,The contents of the whole study were summarized: the efficient extraction of the effective components of Phoebe sheareri,the preliminary screening of the active part of Phoebe sheareri,the biological activity evaluation of each polar part of Phoebe sheareri,separation,purification and identification of active ingredients.In the second part,based on the extraction of ethanol solvent,the single factor control and Response surface analysis method were used to optimize the extraction process of ethanol and determine the optimum extraction conditions.The results showed that the effects of ethanol concentration,extraction time and extraction temperature on the extraction rate of Phoebe sheareri were extremely significant,and the extraction timehad the greatest effect and the ethanol concentration was the second.The optimum extraction conditions were as follows:ethanol concentration 95%,extraction time 6h,extraction temperature 90 ?,ratio of material to liquid1: 8,under this condition,the extraction rate was 6.6%.In the third part,based on the efficient extraction of Phoebe sheareri.The extraction then portioined with petroleum ether,ethylacetate and n-butanol respectively which yielded petroleum ether fraction,ethylacetate fraction and n-butanol fraction.A series of experiments were carried out to study the bioactivities of four extractions,Different models(antioxidant activities,inhibiting the growth of microorganisms and inhibiting the proliferation of tumor cells)were used.The results showed that the 95%ethanol extract and ethyl acetate fraction were found to have significant antioxidant activity.While the antibacterial activity of the extract from the petroleum ether was obvious in particular to inhibit the food-borne microbial(Escherichia coli,Salmonella,Staphylococcus aureus)growth effect.In order to inhibit the proliferation of tumor cells,SPCA-1,MCF-7and HepG2 were used as the research object.It was found that the effects of the extraction of Phoebe sheareri on tumor cell growth were not obvious,Ester part of the three tumor cells have a certain growth inhibition.And the part of the petroleum ether of Phoebe sheareri has almost no cytotoxic activity,and the partial nicotine fraction of Phoebe sheareri shows selective cytotoxicity to human breast cancer cell MCF-7.In the fourth part,the extraction of Phoebe sheareri rhizome and the volatile components of Phoebe sheareri leaves were analyzed by liquid phase-tandem mass spectrometry(LC-MSMS)and gas-phase tandem mass spectrometry(GC-MSMS).The results showed that 12 compounds were identified.It include Tetraethyl-o-scutellarin,aconitine,catechins,L-epicatechin,Miconioside A,Linoleic acid,octadecatrienoic acid,Machilusolide D,iso-ferulic acid,Myristyl-acetate.The compounds were identified by GC-MSMS,and 19 compounds were identified.Thesecompounds mainly include compounds such as olefins,esters,alkanes,alcohols and aldehydes.In the fifth part,Ethyl acetate fraction of Phoebe sheareri was separated and purified by various chromatography(thin layer chromatography and various column chromatography include silica gel column chromatographyand so on).Combined with modern separation and purification methods,semi-preparative liquid phase and high performance liquid chromatography.Four compounds were obtained and their structure was identified by mass spectrometry and nuclear magnetic resonance as oleanolic acid,syringareinol,ferulic acid,catechins respectively.