HPLC electrochemical fluorometric detection of amino acids including tryptophan using 4-fluoro-7-nitrobenzo-2-oxa-1,3-diazole

Biomed Chromatogr. 1987;2(3):99-103. doi: 10.1002/bmc.1130020304.

Abstract

It has been shown that by electrochemical oxidation 7-nitrobenzo-2-oxa-1,3-diazole-tryptophan (NBD-T) is converted to fluorophores having the same emission and excitation spectra as those for other NBD-amino acids. NBD-dioxindolylalanine was tentatively assumed to be a main electrochemical oxidation product of NBD-tryptophan. A coulochemical cell placed between an analytical column and a fluorometer showed no detrimental effect on the separation of NBD-amino acids by reversed phase HPLC. Highly sensitive fluorescence detection was achieved for amino and imino acids at 10-100 fmol levels. The detection limit for tryptophan was 50 fmol.

MeSH terms

  • 4-Chloro-7-nitrobenzofurazan* / analogs & derivatives
  • Amino Acids / analysis*
  • Chromatography, High Pressure Liquid / methods
  • Electrochemistry
  • Fluorescent Dyes*
  • Fluorometry
  • Oxadiazoles* / analogs & derivatives
  • Tryptophan / analysis*

Substances

  • Amino Acids
  • Fluorescent Dyes
  • Oxadiazoles
  • 7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole
  • Tryptophan
  • 4-Chloro-7-nitrobenzofurazan