Liquid chromatography method for detecting native fluorescent bioamines in urine using post-column derivatization and intramolecular FRET detection.
Liquid chromatography (LC) with fluorescence detection is described for simultaneous determination of native fluorescent bioamines (indoleamines and catecholamines). This is based on intramolecular fluorescence resonance energy transfer (FRET) in an LC system following post-column derivatization of native fluorescent bioamines' amino groups with o-phthalaldehyde (OPA). OPA fluorescence was achieved through an intramolecular FRET process when the molecules were excited at maximum excitation wavelength of the native fluorescent bioamines. Bioamines separated by reversed-phase LC on ODS column were derivatized with OPA and 2-mercaptoethanol. This method provides sufficient selectivity and sensitivity for the determination of normetanephrine, dopamine, tyrosine, 5-hydroxytryptamine, tryptamine, and tryptophan in healthy human urine without prior sample purification.
Yoshitake
Makoto
M
Faculty of Pharmaceutical Sciences, Fukuoka University, Nanakuma, Johnan, Fukuoka 814-0180, Japan.
Nohta
Hitoshi
H
Ogata
Susumu
S
Todoroki
Kenichiro
K
Yoshida
Hideyuki
H
Yoshitake
Takashi
T
Yamaguchi
Masatoshi
M
D001679:Biogenic Amines / Q000737:chemistry / Q000652:urine*; D002395:Catecholamines / Q000737:chemistry / Q000652:urine; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D004298:Dopamine / Q000737:chemistry / Q000652:urine; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006801:Humans; D009647:Normetanephrine / Q000737:chemistry / Q000652:urine; D015203:Reproducibility of Results; D014364:Tryptophan / Q000737:chemistry / Q000652:urine; D014439:Tyramine / Q000737:chemistry / Q000652:urine; D014443:Tyrosine / Q000737:chemistry / Q000652:urine; D009764:o-Phthalaldehyde / Q000737:chemistry / Q000652:urine
D001679:Biogenic Amines; D002395:Catecholamines; D009647:Normetanephrine; D014443:Tyrosine; D009764:o-Phthalaldehyde; D014364:Tryptophan; D004298:Dopamine; D014439:Tyramine