RP-HPLRC Separation of Polyamines After Automatic FMOC-CI Derivatization and Precolumn Sample Clean-Up Using Column Switching

Abstract

A fully automated RP-HPLC procedure including precolumn derivatization and sample clean-up was developed for the separation and determination of polyamines in plant tissue. The crude plant extract was derivatized with FMOC-CI during 3 min without a prechromatographic purification step. The sample matrix, the reagent excess and most of the reaction by-products were effectively removed by a simple end-cut technique using column switching. The FMOC-polyamines were separated through an ODS microbore column with good resolution and detected with a fluorescence detector. The detection limit was less than 40 femtomoles for all analysed FMOC-polyamines.