6,7-Dimethoxy-1-methyl-2(1H)-quinoxalinone-3-propionylcarboxylic acid hydrazide: a highly sensitive fluorescence derivatisation reagent for carboxylic acids in high-performance liquid chromatography

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Abstract
6,7-Dimethoxy-1-methyl-2(1H)-quinoxalinone-3-propionylcarboxylic acid hydrazide was found to be a highly sensitive fluorescence derivatisation reagent for carboxylic acids in high-performance liquid chromatography. The reaction conditions were optimised for various C5–C20 saturated fatty acids. The reagent readily reacted with the fatty acids in aqueous solution in the presence of pyridine and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide at room temperature to produce the corresponding fluorescent derivatives, which were separated on a reversed-phase column, Biofine RPC-SC 18 B, by gradient elution with 40–100% aqueous methanol. The derivatives were detected spectrofluorimetrically at 447 nm with excitation at 365 nm. The detection limits (signal to noise ratio = 3) for the acids were 3–6 fmol for an injection volume of 10 µl. The reagent was also applied to the derivatisation of some metabolites of arachidonic acid.

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