Abstract
3-Bromomethyl-6,7-dimethoxy-1-methyl-2(1 H)-quinoxalinone was found to be a selective and highly sensitive fluorescence derivatization reagent for carboxylic acids in high-performance liquid chromatography. Its reactivity was investigated for various linear C 3C 20 saturated fatty acids. The reagent reacts with the fatty acids in acetonitrile in the presence of 18-crown-6 and potassium carbonate to produce the corresponding fluorescent esters, which can be separated on a reversed-phase column, Radial-Pak C 18 cartridge, with gradient elution using 57–100% (v/v) aqueous methanol; the detection limits for the acids were 0.3–1 fmol for an injection volume of 5 μl. The reagent also reacts with unsaturated fatty, dicarboxylic, aromatic carboxylic and hydroxycarboxylic acids and acidic nucleotides to form fluorescent derivatives. α-Keto acids and α-amino acids do not give fluorescent derivatives under these conditions.
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