Isolation And NMR Evaluation Of Anthocyanins From Mulberry And Flavonoid Glycoside From Passiflora

From Tang dynasty on mulberry have being used as tonics by Chinese for its Ziyin quality,the effectiveness of the blood,promoting saliva and body fluids moistens dry areas. It can be used to cure swimmy tinnitus,physically and mentally fatigued,early graying hair,thirst,diabetes with heat,diarrhea with blood deficient etc. Morden reserches showed that anthocyanins play the main role in psysiologocal activities of mulberry, therefore we tried to isolate anthocyanin monomer. this study is supposed to provides the foundation of systematically studying on the psysiologocal activities and other properties of anthocyanins from mulberry and to provides the theoritical directions to correlative food industry.Mulberry fruits were extracted adequately with methnol contained 0.1% hydrochloric acid. Then macropore absorption resin AB-8 was used to purify anthocyanins extractive. Preparative high performance liquid chromatography was applied to isolate anthocyanins from mulberry. Two collected fractions were concentrated by rotary evaporator then Vacuum freeze drier.Structures were evaluated by 'H nuclear magnetic resonance and reference. The result showed that anthocyanin 1 is cyaniding-3-O-glucoside; anthocyanin 2 is cyanidin-3-O-(6"-O-a-hhamnoPyrano-side-p-D-galacopyranoside).A new degredation product would generate if anthocyanin sample degredated fully under no protective measures. We purified this compound by HPLC using the solvents CH3OH (A) and H2O. After twice purification we got a high purified conpound (96.0%).'H-NMR and 13C-NMR evaluated this compound as 5-(hydroxymethyl)-2-furaldehyde.P. edulis f.flavicarpa Degener, a species in Passiflora genus, is widespread throughout the tropics and subtropics. The chemical investigation of P. edulis f.flavicarpa Degener afforded the isolation of different classes of natural products including flavonoids and alkaloids. The flavonoids is a natural antioxidant, scavengening free radicle, anti-ageing, improving immunity etc. Some reserches published in the past few years found that Passiflora showed anti-cancer,antianxiety,calm,antibiosis,anti-inflammation and so on. Thus, we used high-speed counter-current chromatography to isolate flavone C-glycosides to clarify which constituents is responsible for its activities.Reports on the flavonoid glycosides isolation applied the High speed counter current chromatography, have not been found. The leaves of P. edulis f.flavicarpa Degener were extracted by maceration using a mixture of MeOH-H2O (3:2) at room temperature. The organic extract was purified by column chromatography on polyamide firstly. The aquous phase fraction was collected and concentrated. Then the recycling high-speed counter-current chromatography (HSCCC) was used to separate from crude sample with a solvent system composed of n-butanol-ethyl acetate-water(4:1:5). At last, the luteolin-6,8-glucopyranoside(7.3mg) and apigenin-6,8-glucopyranoside(14.4mg) were completely separated by recycling HSCCC preparation with one recycle. The contents were 95.6%,96,3% and the recovery rates were 71.87%,76.31%, respectively. Compared with the chromatography,the resolution of the difficult separated compounds was enhanced by the recycling HSCCC. So the recycling HSCCC provides a convenient and rapid way to the difficult separated compounds.