HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC DETERMINATION OF CATECHOLAMINES AND THEIR PRECURSOR AND METABOLITES IN HUMAN URINE AND PLASMA BY POSTCOLUMN DERIVATIZATION INVOLVING CHEMICAL OXIDATION FOLLOWED BY FLUORESCENCE REACTION

A high-performance liquid chromatographic method for the determination of catecholamines and their precursor and metabolites [amino compounds (norepinephrine, epinephrine, dopamine, normetanephrine, metanephrine, 3-methoxytyramine, and l-DOPA), acidic compounds (3,4-dihydroxyphenylacetic acid, vanillylmandelic acid, and homovanillic acid), and alcoholic compound ((4-hydroxy-3-methoxyphenyl)ethylene glycol)] in human urine and plasma. Urine and plasma samples deproteinized with perchloric acid in the presence of isoproterenol and 3,4-dihydroxyphenylpropanoic acid (internal standards) are fractionated by solid-phase extraction on a strong cation-exchange resin cartridge (Toyopak IC-SP S) into two fractions (amine fraction and acid-alcohol fraction). The compounds in each fraction are separated by an ion-pair reversed-phase chromatography on a TSK gel ODS-80TM with isocratic elution and on-line derivatized by periodate oxidation followed by a fluorescence reaction using meso-1,2-diphenylethylenediamine. The detection limits ( S N = 5) vary from 0.5 to 95 pmol/ml, depending on the compounds. © 1992.