Determination of Catecholamines and Indoleamines in Human Urine Based on Intramolecular Excimer-forming Derivatization and Fluorescence Detection

YOSHIDA Hideyuki, KIDO Fumito, YOSHITAKE Makoto, TODOROKI Kenichiro, NOHTA Hitoshi, YAMAGUCHI Masatoshi

doi:10.2116/analsci.23.485

Determination of Catecholamines and Indoleamines in Human Urine Based on Intramolecular Excimer-forming Derivatization and Fluorescence Detection

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YOSHIDA Hideyuki

Faculty of Pharmaceutical Sciences, Fukuoka University
KIDO Fumito

Faculty of Pharmaceutical Sciences, Fukuoka University
YOSHITAKE Makoto

Faculty of Pharmaceutical Sciences, Fukuoka University
TODOROKI Kenichiro

Faculty of Pharmaceutical Sciences, Fukuoka University
NOHTA Hitoshi

Faculty of Pharmaceutical Sciences, Fukuoka University
YAMAGUCHI Masatoshi

Faculty of Pharmaceutical Sciences, Fukuoka University

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抄録

A liquid chromatographic (LC) determination of catecholamines and indoleamines is described. This is based on intramolecular excimer-forming fluorescence derivatization with 4-(1-pyrene)butanoyl chloride, followed by reversed-phase LC. The analytes, containing an amino moiety and phenolic hydroxyl moieties in a molecule, were converted to the corresponding polypyrene-labeled derivatives by one-step derivatization. They afforded intramolecular excimer fluorescence, which can clearly be discriminated from the normal fluorescence emitted from reagent blanks. The detection limits (S/N = 3) for catecholamines and indoleamines were femto-mole levels per 20-µL injection. Furthermore, this method was applied to a urine assay.