Abstract
An on-column fluorometric derivatization method has been developed for the determination of polyamines using high-performance liquid chromatography. The system for the derivatization consisted of a single-plunger pump and a reversed-phase C18 polymer column with a mobile phase of acetonitrile and alkaline borate buffer solution containing o-phthalaldehyde and N-acetyl-L-cysteine as derivatizing reagents. The polyamines injected onto the liquid
chromatograph are derivatized to fluorophores at the inlet of the column, followed by chromatography on the same column. Optimization of the liquid chromatography conditions led to a simple analytical method for the simultaneous determination of polyamines with high reproducibility and linearity. Application of this method to the determination of polyamines in food samples resulted in overall mean recoveries greater than 85% at fortification values of 20-200 μg g-1 of each polyamine.
