Recently, it was reported that polyamines, such as spermidine and spermine, play an important role in the inhibition of Leukocyte Function Antigen-1 (LFA-1) related to the induction of inflammation, immune function, and the arteriosclerosis process. 8,9 Since polyamines orally taken are directly absorbed, more attention has been given to food rich in polyamines for the possibility of controlling arteriosclerosis.Both pre-column and post-column derivatization methods for amine analyses have been extensively used in HPLC to improve the sensitivity and selectivity.10-17 A post-column derivatization method is the much preferred method for amines, because it offers easy operation, less sample manipulation, and high 2013 © The Japan Society for Analytical Chemistry † To whom correspondence should be addressed. 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.

In this study, using a single solvent system (30 mM sodium

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