Preparative isolation and purification of seven main antioxidants from Eucommia ulmoides Oliv. (Du-zhong) leaves using HSCCC guided by DPPH-HPLC experiment

Food Chem. 2013 Aug 15;139(1-4):563-70. doi: 10.1016/j.foodchem.2013.02.006. Epub 2013 Feb 16.


Seven antioxidants were purified from Eucommia ulmoides Oliv. leaves using HSCCC guided by DPPH-HPLC experiment. HSCCC was successfully used to separate target antioxidants by three runs with different solvent systems after D101 column chromatography fractionation. Ethyl acetate-n-butanol-water (1:2:3, v/v/v) was selected as the optimum solvent system to purify geniposidic acid. Ethyl acetate-ethanol-water (4:1:5, v/v/v) was used to isolate caffeic acid, chlorogenic acid and ferulic acid. While three flavonoids, quercetin-3-O-sambubioside, rutin and isoquercitrin were purified by petroleum ether-ethyl acetate-methanol-water (1:5:1:5, v/v/v/v). The structures were identified by MS and NMR. Antioxidant activities were assessed, and compounds 2-7 showed strong antioxidant activities. This is the first report about separation of antioxidants from E. ulmoides leaves by HSCCC. The results indicated that the combinative methods using DPPH-HPLC and HSCCC could be widely applied for screening and isolation of antioxidants from complex extracts.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antioxidants / chemistry*
  • Antioxidants / isolation & purification*
  • Biphenyl Compounds
  • Chromatography, High Pressure Liquid / methods*
  • Countercurrent Distribution / methods*
  • Eucommiaceae / chemistry*
  • Picrates
  • Plant Extracts / chemistry*
  • Plant Extracts / isolation & purification*
  • Plant Leaves / chemistry


  • Antioxidants
  • Biphenyl Compounds
  • Picrates
  • Plant Extracts
  • 1,1-diphenyl-2-picrylhydrazyl