Add time:07/30/2019         Source:sciencedirect.com

A sytem for the high-performance liquid chromatography of carboxylic acids using 4-bromomethyl-7-acetoxycoumarin (Br-Mac) as the fluorescence reagents is described. Br-Mac reacts with carboxylic acids to give the ester derivatives, which are separated using reversed-phase liquid chromatography. The eluate from the column is mixed with an alkaline solution. The labelled carboxylic acids are hydrolysed to the fluorescent coumarin derivatives, which are introduced into a flow-through fluorimeter. In this system, the fluorescent hydrolysate, equimolar to a carboxylic acid, is detected and this fluorophor is common to every carboxylic acid. Only a slight variation is found in the peak areas for various carboxylic acids. A gradient elution technique is effectively used in this system because the fluorescence quantum yield of the fluorescent hydrolysate is not affected by the constitution of the mobile phase. Low femtomole levels of carboxylic acids can be detected.

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