Determination of fluorescent cyanobenz[f]isoindole derivatives of dopamine and norepinephrine using high performance liquid chromatography with chemiluminescence detection.
Dopamine (DA), norepinephrine (NE) and 3,4-dihydroxybenzylamine (DHBA) are converted to highly fluorescent cyanobenz[f]isoindole (CBI) derivatives by reacting these amines with naphthalene-2,3-dicarboxaldehyde in the presence of cyanide ion. Femtomole amounts of these CBI derivatives separated by reverse-phase high performance liquid chromatography can be detected by a post-column chemiluminescence system utilizing bis(2,4-dinitrophenyl) oxalate and hydrogen peroxide. Linear detection response was observed between 1 to 600 fmol, giving a signal to noise ratio of approximately 15 at the 1 fmol level. An assay procedure was developed for the determination of DA and NE in 20 microL of urine sample using DHBA as the internal standard.
Kawasaki
T
T
Center for Bioanalytical Research, University of Kansas, Lawrence 66045.
Imai
K
K
Higuchi
T
T
Wong
O S
OS
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004140:Dinitrophenols; D004298:Dopamine / Q000378:metabolism* / Q000652:urine; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007211:Indoles / Q000032:analysis* / Q000378:metabolism; D008163:Luminescent Measurements; D009570:Nitriles / Q000032:analysis* / Q000378:metabolism; D009638:Norepinephrine / Q000378:metabolism* / Q000652:urine; D010070:Oxalates / Q000494:pharmacology; D013997:Time Factors
D004140:Dinitrophenols; D007211:Indoles; D009570:Nitriles; D010070:Oxalates; C041019:bis(2,4-dinitrophenyl) oxalate; D006861:Hydrogen Peroxide; D004298:Dopamine; D009638:Norepinephrine