Analysis of ascorbate in plant tissues by high-performance capillary zone electrophoresis

Anal Biochem. 1996 Jul 15;239(1):8-19. doi: 10.1006/abio.1996.0284.


We describe here a simple and rapid capillary electrophoresis method for the determination of ascorbic acid (L-AA) and isoascorbic acid (D-AA) in vegetative tissues. For optimal yields and stabilization, samples are extracted with cold 3% metaphosphoric acid. Hydrophobic contaminants are then removed by passage through a C18 solid-phase extraction cartridge. The analysis itself is performed on a fused silica capillary with 200 mM borate, pH 9, as the carrier electrolyte, using on-line diode array detection over the range 190-350 nm. Quantitation was performed at 260 nm, the uv-absorption maximum for ascorbate at this pH. This method has a minimum detection limit of 84 fmol/injection and linearity of detector response was observed up to at least 12 pmol/injection. We also describe the influence of electrolyte concentration, pH, and the presence of detergent on separations of L-AA, D-AA, and L-galacturonic acid-1,4-lactone. The protocol has been demonstrated to be suitable for the analysis of L-AA in Arabidopsis, parsley, and mushroom. The method has superior resolution to comparable HPLC separations, a comparable analysis time, but lower sensitivity because of the concentration limitations of the detection system.

Publication types

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

MeSH terms

  • Arabidopsis / chemistry
  • Ascorbate Oxidase / pharmacology
  • Ascorbic Acid / analysis*
  • Basidiomycota / chemistry
  • Chromatography, High Pressure Liquid
  • Electrophoresis, Capillary / methods*
  • Hydrogen-Ion Concentration
  • Plants / chemistry*
  • Reproducibility of Results
  • Sodium Dodecyl Sulfate / pharmacology


  • isoascorbic acid
  • Sodium Dodecyl Sulfate
  • Ascorbate Oxidase
  • Ascorbic Acid