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

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Abstract
It has been shown that by electrochemical oxidation 7‐nitrobenzo‐2‐oxa‐1,3‐diazole‐tryptochan (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.

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