The Research On The Isolation And Identification Of The Catalitaxol And Its Analogue

Three unknown compounds were measured from the endophytic fungi FSN002 by HPLC(High Performance Liquid Chromatography).The compounds were speculated to be catalitaxol(RT 21 min and ?max 282 nm),the acid degradation product(RT 24 min and ?max 306 nm)and the yellow degradation product(RT 28 min and ?max 362 nm).Specifically,the yellow natural product catalitaxol was reported to treat liver cancer and brain cancer without damaging other normal body cells.To better measure catalitaxol by HPLC,the inorganic salt Na Cl was added into the fermentation liquor.To isolate and identify the three unkown compounds,resin adsorption separation,extraction,thin layer chromatography,LC-MS(liquid chromatography-mass spectrometry)and NMR(nuclear magnetic resonance)were used in this study.The main results were as follows:(1)The resinous material ADS-7 was chosen for the isolation of catalitaxol.Catalitaxol was purified to the salt methanol solution through the cleaning process containing 4 steps: saturated Na C-water-methanol-saturated methanol.At the same time,the hydrophilic pigment in the fermentation liquid was removed.Besides,the compound 3 was separated and purified through a combined method of polyamide,thin layer chromatography and preparative ODS column analysis.The chromatographic purity of compound 3 was 91.54%.Subsequently,the purified compound 3 was identified by NMR and LC-MS analysis.These results show that the molecular weight of compound 3 is 496.21 g/mol,and the molecular formula is C28H32O8.The compound 3 is Trichodimerol,which belongs to the polyketide.Its space structure is symmetrical,and chemical properties are similar to the flavonoids.Trichodimerol with the antibacterial and antifungal function is extracted from both the mangrove endophytic fungus Trichoderma sp.and Hypoxylon oceanicum.(2)Catalitaxol was separated and purified from the fermentation liquor using liquid-liquid extraction under the acid and alkaline conditions.The separation and purification method was established through the type of solvents,the ratios of the solvent to the material(by volume),the p H conditions of the extraction system,and detected by HPLC.The partition coefficients of oil-water in four different solvent decrease in order of ethyl acetate,chloroform,dichloromethane and methylbenzene.Reducing the extraction volume resulted in an degradation of catalitaxol.When the p H was increased,the oil-water distribution coefficient is decreased.The extraction rate of catalitaxol was 78.28% using fermentation liquor and ethyl acetate mixed solvent(1:1),p H= 4.5 and sodium hydroxide and ethyl acetate mixed solvent(1:3),p H= 9.The final chromatographic purity of product is 97.7%.The liquid-liquid extraction under the acid and alkaline conditions can be used to separate and purify the catalitaxol.