Separation and purification of glucosinolates from crude plant homogenates by high-speed counter-current chromatography

Jed W. Fahey, Kristina L. Wade, Katherine K. Stephenson, F. Edward Chou

Research output: Contribution to journalArticlepeer-review

65 Scopus citations


Glucosinolates are anionic, hydrophilic plant secondary metabolites which are of particular interest due to their role in the prevention of cancer and other chronic and degenerative diseases. The separation and purification of glucosinolates from a variety of plant sources (e.g. seeds of broccoli, arugula and the horseradish tree), was achieved using high-speed counter-current chromatography (HSCCC). A high-salt, highly polar system containing 1-propanol-acetonitrile-saturated aqueous ammonium sulfate-water (1:0.5:1.2:1), was run on a semi-preparative scale and then transferred directly to preparative scale. Up to 7 g of a concentrated methanolic syrup containing about 10% glucosinolates was loaded on an 850-ml HSCCC column, and good separation and recovery were demonstrated for 4-methylsulfinylbutyl, 3-methylsulfinylpropyl, 4-methylthiobutyl, 2-propenyl and 4-(rhamnopyranosyloxy)benzyl glucosinolates. Multiple injections (5 to 6 times) were performed with well-preserved liquid stationary phase under centrifugal force. Pooled sequential runs with broccoli seed extract yielded about 20 g of its predominant glucosinolate, glucoraphanin, which was produced at >95% purity and reduced to powdered form.

Original languageEnglish (US)
Pages (from-to)85-93
Number of pages9
JournalJournal of Chromatography A
Issue number1-2
StatePublished - May 9 2003
Externally publishedYes


  • Brassica spp.
  • Counter-current chromatography
  • Glucoraphanin
  • Glucosinolates
  • Plant materials
  • Vegetables

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry


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