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Preparative isolation and purification of antioxidative stilbene oligomers from Vitis chunganeniss using high-speed counter-current chromatography in stepwise elution mode
- He, Shan, Lu, Yanbin, Jiang, Liyan, Wu, Bin, Zhang, Feiying, Pan, Yuanjiang
- Journal of separation science 2009 v.32 no.14 pp. 2339-2345
- Vitis, antioxidant activity, countercurrent chromatography, high performance liquid chromatography, nuclear magnetic resonance spectroscopy, singlet oxygen
- Preparative high-speed counter-current chromatography (HSCCC) was successfully applied to the isolation and purification of three stilbene oligomers from Vitis chunganeniss using stepwise elution with a pair of two-phase solvent systems composed of n-hexane-ethyl acetate-methanol-water at (2:5:2:5, v/v) and (1:2:1:2, v/v). The preparative HSCCC separation was performed on 800 mg of crude sample yielding hopeaphenol (21.1 mg), amurensin G (37.2 mg) and vitisin A (95.6 mg) in a one-step separation, with purities over 95% as determined by HPLC. The structures of these three compounds were identified by MS, ¹H NMR and ¹³C NMR. In addition, their antioxidant activities were screened by DPPH assay, where vitisin A showed strong antioxidant activity. Further EPR experiments with spin-trapping technique demonstrated that vitisin A is a potent and selective singlet oxygen quencher, which may be used in singlet oxygen-mediated diseases as a pharmacological agent.