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, vanillyl-mandelic 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.
Jeon
H K
HK
Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Nohta
H
H
Ohkura
Y
Y
D000328:Adult; D002395:Catecholamines / Q000097:blood / Q000378:metabolism* / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004298:Dopamine / Q000097:blood / Q000378:metabolism* / Q000652:urine; D004837:Epinephrine / Q000097:blood / Q000378:metabolism* / Q000652:urine; D005260:Female; D006801:Humans; D007202:Indicators and Reagents; D008297:Male; D009638:Norepinephrine / Q000097:blood / Q000378:metabolism* / Q000652:urine; D010084:Oxidation-Reduction; D012016:Reference Values; D013050:Spectrometry, Fluorescence / Q000379:methods
D002395:Catecholamines; D007202:Indicators and Reagents; D004298:Dopamine; D009638:Norepinephrine; D004837:Epinephrine