4-Dimethylaminobenzylamine as a sensitive chemiluminescence derivatization reagent for 5-hydroxyindoles and its application to their quantification in human platelet-poor plasma.
A selective and sensitive high-performance liquid chromatographic method with chemiluminescence detection for the determination of 5-hydroxyindoles is described, based on the reaction of 5-hydroxyindoles with 4-dimethylaminobenzylamine. Serotonin, 5-hydroxyindole-3-acetic acid, 5-hydroxytryptophol, 5-hydroxyindole-3-acetamide and N-acetyl-5-hydroxytryptamine were used as model compounds to optimize the derivatization and chemiluminescent reaction. The reagent reacts with 5-hydroxyindoles in slightly alkaline media in the presence of potassium hexacyanoferrate(III) to give the corresponding derivatives, which can be separated on a reversed-phase column, Wakosil-II 5C18RS, with aqueous acetonitrile as an eluent. The derivatives were detected by peroxyoxalate chemiluminescence detection. The detection limits are in the range of 0.5-1.2 fmol per 100-microl injection. The method was applied to the simultaneous determination of serotonin and 5-hydroxyindole-3-acetic acid in human platelet-poor plasma.
Ishida
J
J
Faculty of Pharmaceutical Sciences, Fukuoka University, Japan.
Takada
M
M
Hitoshi
N
N
Iizuka
R
R
Yamaguchi
M
M
D000814:Aniline Compounds / Q000737:chemistry*; D001596:Benzylamines / Q000737:chemistry*; D001792:Blood Platelets; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D006897:Hydroxyindoleacetic Acid / Q000097:blood; D007202:Indicators and Reagents / Q000737:chemistry*; D007211:Indoles / Q000097:blood*; D008163:Luminescent Measurements; D010949:Plasma / Q000737:chemistry; D015203:Reproducibility of Results; D012701:Serotonin / Q000097:blood; D013050:Spectrometry, Fluorescence
D000814:Aniline Compounds; D001596:Benzylamines; D007202:Indicators and Reagents; D007211:Indoles; C035815:4-dimethylaminobenzylamine; C026785:5-hydroxyindole; D012701:Serotonin; D006897:Hydroxyindoleacetic Acid