Fluorimetric determination of kynurenine in human serum by high-performance liquid chromatography coupled with post-column photochemical reaction with hydrogen peroxide

A high-performance liquid chromatographic method involving post-column photochemical reaction and fluorimetric detection has been developed for the determination of kynurenine in serum. Kynurenine was separated on a column of Capcell Pak C18 (resistant to pH 10). The mobile phase consisted of 0.05 M Na2B4O7-0.1 M KH2PO4 buffer (pH 8.5)-ethanol (97:3, v/v) containing 60 mM hydrogen peroxide. The post-column reagent, containing 60% (v/v) ethanol, was mixed with the mobile phase, which was irradiated with ultraviolet light to induce fluorescence. The recovery of kynurenine was 95.9 +/- 5.0% (n = 6). The method allows the determination of as little as 2 pmol of kynurenine.