Determination of polyamines in biological materials by HPLC with 9-fluorenylmethyl chloroformate precolumn derivatization

Di- and polyamines have been determined in the femtomol range by liquid chromatography on a reversed-phase C8 column with a linear gradient of acetate buffer/acetonitrile as mobile phase and fluorimetric detection at 265/310 nm after simple derivatization with 9-fluorenylmethyl chloroformate (FMOC) within 3 min at pH 7.8 in presence of aspartic acid to remove excess reagent. The FMOC—polyamine-derivatives have been stable in a methanol—water-solution for at least 10 h. The method can be directly applied to crude plant extracts, and it is not interfered by carbohydrates and phenolics.