Separation And Composition Analysis Of Triterpenoids From Pyrus Ussuriensis Maxim Cv. Anli

Pyrus ussuriensis Maxim cv. Anli belongs to the Pyrus genus of Rosaceasefamily. To lay a foundation for development of P. ussuriensis Maxim cv. Anli, basedon P. ussuriensis Maxim cv. Anli as raw materials, this paper mainly studiedtriterpenoids in the extraction and separation, structural characterization andphysiological activity. The main results were as follows:1. Extraction and separationCompared with heat reflux extraction and macro-wave assisted extraction, theoptimal conditions of the pressurized liquid extraction from P. ussuriensis Maxim cv.Anli in total triterpenoids included heating temperature of85℃, extraction time of4min,3times of circulation, and55%ethanol concentration. Under these conditions，the extraction rate was up to8.62%.Silica gel and macroporous resin column chromatography were used forpreliminary separation and purification P. ussuriensis Maxim cv. Anli extracts, andSephadex LH-20column chromatography and HPLC were used for furtherpurification.2. Structural analysisBased on the analysis of LC-MS, there are16kinds of triterpenoids ingredientsand12kinds of flavonoids ingredients. The triterpenoids ingredients are as follows:Roxburic acid, Pomonic acid, Alphitolic acid, Oleanic acid, Ursolic Acid, Oleanonicacid,7-dehydroavenasterol, Stigmasta-4,22-dien-3-one, Betulin, Glochidone, Ganoderol A, Nebrosteroid D, Nebrosteroid E and Desmosterol. The flavonoids ingredientsare as follows: luteolin7-O-rutinoside, kaempferol3-O-rhamnosylgalactoside, Kaempferol-3-O-glucoside, Luteolin7-O-glucoside, Luteolin4’-O-glucoside, Apigenin7-O-glucoside, Apigenin4’-O-glucoside, Isorhamnetin3-O-galactoside, Isorhamnetin3-O-glucoside, Isorhamnetin3-O-rhamnosylgalactoside, Isorhamnetin3-O-rhamnosylglucoside, Chrysoeriol7-O-glucoside.11kinds of triterpenoids ingredients were foundin the Pyrus genus for the first time.3. Research on physiological activity DPPH method is used to antioxidant activity study of P. ussuriensis Maxim cv.Anli. Ethyl acetate parts of the extracts on DPPH free radical maximum clearance ratewas60.47%, and n-butanol parts of extracts on DPPH free radical clearance rate was51.98%. The experiment proved that ethyl acetate extract has stronger antioxidantactivity.Through sulfonyl rhodamine B (sulforhodamine B, SRB) protein stainingmethod for screening of antitumor activity of P. ussuriensis Maxim (cv. Anli. Whensample concentration of1mg/ml, ethyl acetate parts of the extract of human livercancer cell Bel-7402growth inhibition rate was as high as66.3%, and n-butanol partextracts of human liver cancer cells Bel-7402growth inhibition rate was32.7%. Theexperiment proved that ethyl acetate parts of extracts on the growth of humanhepatocellular carcinoma cells Bel-7402had stronger inhibitory activity.