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Coupled-column liquid chromatographic analysis of catecholamines, serotonin, and metabolites in human urine
T J Panholzer1, J Beyer, K Lichtwald
1University Hospital of Mainz, Department of Internal Medicine/Endocrin ology and Metablic Diseases, D-5101, Mainz, Germany.
Abstract:
A column-switching HPLC system was utilized for the simultaneous determination of epinephrine, norepinephrine, dopamine, serotonin, and their metabolites metanephrine, normetanephrine, 3, 4-dihydroxyphenylacetic acid, homovanillic acid, and 5-hydroxyindoleacetic acid in human urine. The sample was injected directly onto a C18-alkyl-diol silica precolumn, which separated the analytes from matrix. The analytes were eluted from the precolumn onto the analytical column by the use of column-switching techniques and were then separated on the analytical column by means of ion-pair reversed-phase HPLC. The analytes were then oxidized to the corresponding quinones and converted into fluorescent derivatives by reaction with meso-1,2-diphenylethylenediamine.