title: Optimization of an online post-column derivatization system for ultra high-performance liquid chromatography (UHPLC) and its applications to analysis of biogenic amines.
abstract: We investigated the efficacy and utility of a post-column derivatization method for the detection of amines with o-phthalaldehyde (OPA) in ultra high-performance liquid chromatography (UHPLC). Since it was anticipated that a derivatization reaction system placed downstream of the column would give considerable band broadening to narrow peaks generated by a UHPLC column, we examined the contributions of the dimensions of the reaction tube, the fluorescence (FL) detector flow cell, and the column to the extra-column peak broadening, and optimized the post-column derivatization system in UHPLC. We successfully separated amines within 6 min with gradient elution by using the post-column derivatization system connected to a UHPLC system. The analysis time was reduced by a factor of 7 compared with that by conventional HPLC. The detection limits were 18.0 - 105.7 fmol, depending on the compounds; the reproducibilities were 0.11 - 0.82% RSD for the retention times and 0.78 - 1.66% RSD for the peak areas (n = 10), respectively. The linear dynamic ranges were up to 12, 20, and 40 pmol/μL, depending on the compounds, respectively.
authors.affiliation: LC and SFC Applications Laboratory, JASCO Corporation, 2967-5, Ishikawamachi, Hachioji, Tokyo 192-8537, Japan. satoe.iijima@jasco.co.jp
authors.forename: Satoe
authors.initials: S
authors.lastname: Iijima
authors.forename: Yasuyo
authors.initials: Y
authors.lastname: Sato
authors.forename: Masao
authors.initials: M
authors.lastname: Bounoshita
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Miyaji
authors.forename: David J
authors.initials: DJ
authors.lastname: Tognarelli
authors.forename: Muneo
authors.initials: M
authors.lastname: Saito
mesh_terms: D001679:Biogenic Amines / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005504:Food Analysis
chemical_list: D001679:Biogenic Amines