Improved high-performance liquid chromatographic analysis of some carboxylic acids in food and beverages as their p-nitrobenzyl esters.

An improved, straightforward and accurate method for the derivatization and quantitative high-performance liquid chromatographic analysis of carboxylic acids in food and beverages is presented. Thirty-two p-nitrobenzyl esters were prepared in high yields by direct reaction of the free acid with O-(4-nitrobenzyl)-N,N'-diisopropylisourea in dioxane-water (9:1). Benzylmalonic acid was used as a new internal standard. Excess of reagent was removed on a strong cation-exchange resin. The advantage of this original and very convenient procedure over the usual clean-up step involving filtration through a disposable silica cartridge is discussed. The p-nitrobenzyl ester derivatives were readily separated using a single linear gradient of solvent (acetonitrile in water). With a high-performance liquid chromatography column of conventional size (20-25 cm X 4.6 mm I.D.), the analysis lasted for less than 20 min. This analysis time was even shorter (less than 12 min) when using modern small-bore (100 X 2.1 mm I.D.) columns. Applications of the method to the analysis of the main carboxylic acids in coffee, wine and fruit juices are presented.