Analytical Sciences


Abstract − Analytical Sciences, 29(5), 539 (2013).

Optimization of an Online Post-Column Derivatization System for Ultra High-Performance Liquid Chromatography (UHPLC) and Its Applications to Analysis of Biogenic Amines
Satoe IIJIMA,* Yasuyo SATO,* Masao BOUNOSHITA,* Toshihiko MIYAJI,* David J. TOGNARELLI,** and Muneo SAITO*
*LC and SFC Applications Laboratory, JASCO Corporation, 2967-5, Ishikawamachi, Hachioji, Tokyo 192-8537, Japan
**JASCO Incorporated, 28600 Mary’s Court, Easton, MD 21601, USA
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.