title: Selective and sensitive liquid chromatographic determination method of 5-hydroxyindoles with fluorous and fluorogenic derivatization.
abstract: A liquid chromatographic (LC) method with improved selectivity for the simultaneous determination of 5-hydroxyindoles (5-HIs; 5-hydroxytryptophan, 5-hydroxytryptamine, N-acetyl-5-hydroxytryptamine, 5-hydroxyindole-3-acetic acid, and 5-hydroxytryptophol) is described. This method involves precolumn derivatization with 4-(3',3',4',4',5',5',6',6',7',7',8',8',9',9',10',10',10'-heptadecafluorodecyl)benzylamine (HFBA) and separation of the derivatives using a fluorous LC column. In this study, stable benzoxazole derivatives of 5-HIs with HFBA have been obtained by a simple derivatization procedure; their fluorescent properties enabled highly sensitive detection. In addition, only the HFBA derivatives of 5-HIs has been selectively retained on the fluorous LC column via fluorous interaction whereby perfluoroalkyl compounds show affinities with each other, while the non-fluorous compounds did not. The HFBA derivatives were separated within 30 min and the detection limits for 5-HIs in a 20-μL injection volume were 1.2-14 fmol (S/N=3). Furthermore, this method was applied to the analysis of 5-HIs in the human plasma from healthy subjects.
authors.affiliation: Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan, Fukuoka 814-0180, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Sakaguchi
authors.affiliation: Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan, Fukuoka 814-0180, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Ikenaga
authors.affiliation: Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan, Fukuoka 814-0180, Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Yoshida
authors.affiliation: Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan, Fukuoka 814-0180, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Hayama
authors.affiliation: Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan, Fukuoka 814-0180, Japan.
authors.forename: Miki
authors.initials: M
authors.lastname: Itoyama
authors.affiliation: Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan, Fukuoka 814-0180, Japan.
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Todoroki
authors.affiliation: Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan, Fukuoka 814-0180, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Imakyure
authors.affiliation: Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan, Fukuoka 814-0180, Japan.
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Yamaguchi
authors.affiliation: Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan, Fukuoka 814-0180, Japan. Electronic address: nohta@fukuoka-u.ac.jp.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Nohta
mesh_terms: D006916:5-Hydroxytryptophan / Q000032:analysis; D019342:Acetic Acid / Q000032:analysis; D000328:Adult; D001596:Benzylamines / Q000032:analysis; D002853:Chromatography, Liquid / Q000379:methods*; D005260:Female; D006801:Humans; D006897:Hydroxyindoleacetic Acid / Q000032:analysis; D007211:Indoles / Q000032:analysis*; D008297:Male; D015203:Reproducibility of Results; D012701:Serotonin / Q000031:analogs & derivatives / Q000032:analysis; D013050:Spectrometry, Fluorescence / Q000379:methods; D013997:Time Factors; D055815:Young Adult
chemical_list: D001596:Benzylamines; D007211:Indoles; C026785:5-hydroxyindole; D012701:Serotonin; D006897:Hydroxyindoleacetic Acid; D006916:5-Hydroxytryptophan; C006389:N-acetylserotonin; D019342:Acetic Acid
keywords: 5-Hydroxyindoles; Fluorous derivatization; Fluorous separation; Human plasma; Liquid chromatographic-fluorescent detection