Simultaneous determination of acidic 3,4-dihydroxyphenylalanine metabolites and 5-hydroxyindole-3-acetic acid in urine by high-performance liquid chromatography

Abstract We describe a simple and rapid quantitative method for the simultaneous determination of 3,4-dihydroxyphenylalanine acid metabolites and 5-hydroxyindole-3-acetic acid. After solvent extraction from acidified urine, the acids are analyzed by reversed-phase high-performance liquid chromatography. For detection and quantification, we used a fluorescence detector in combination with an amperometric detector to obtain a high degree of specificity. Sample preparation and chromatographic analysis can be completed within an hour. Results by the method correlate well with those by a previously used gas-chromatographic method, but the new method is faster and avoids the need for derivatization.