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Article
Quantification of the production of dihydrokaempferol by Flavanone-3-Hydroxylase using capillary electrophoresis.
Phytochemical Analysis (2002)
  • Daniel K. Owens, East Tennessee State University
  • Tracy Hale, East Tennessee State University
  • Lori J. Wilson
  • Cecilia A. McIntosh, East Tennessee State University
Abstract
A sensitive method using capillary electrophoresis for the separation, detection, and quantification of dihydrokaempferol (1) is reported. Well-resolved, sharp symmetrical peaks were obtained in grapefruit leaf extracts for 1, naringenin (2), and the internal standard, naringin (3). Long columns were required to resolve 1 from 2 in crude enzyme reactions and this resulted in run times of 60 min. The limit of detection for 1 was found to be 1.44 ng/µL (4.2 pg). The method showed excellent linearity and reproducibility. The method was used to determine the activity of flavanone 3-hydroxytransferase (F3H) in leaf tissue of grapefruit by quantification of the production of dihydrokaempferol in controlled time course reactions. The sensitivity of the method makes it adaptable to assaying F3H activity in individual young seedlings and/or in small tissue samples and requires only 100 mg of tissue. Copyright © 2002 John Wiley & Sons, Ltd.
Keywords
  • Capillary electrophoresis,
  • flavanone 3-hydroxytransferase,
  • flavonoids,
  • naringenin,
  • dihydrokaempferol,
  • grapefruit
Publication Date
March 7, 2002
DOI
10.1002/pca.625
Citation Information
Owens, D. K., Hale, T., Wilson, L. J. and McIntosh, C. A. (2002). Quantification of the production of dihydrokaempferol by flavanone 3-hydroxytransferase using capillary electrophoresis. Phytochemical Analysis, 13, 69–74. https://dx.doi.org/10.1002/pca.625