High-performance liquid chromatographic method for the determination of dansyl-polyammines

This paper describes a fast reliable, and a sensitive technique for the separation and quantification of dansylated polyamines by high-performance liquid chromatography. Using a small 33 × 4.6 mm I.D., 3 μm particle size, C18reversed-phase cartridge column and a linear gradient of acetonitrile—heptanesulfonate (10 mM, Ph 3.4), at a flow-rate of 2.5 ml/min, the retention time for different polyamines was: N8-acetyl-spermidine, 1.79 min; N1-acetylspermidine, 1.82 min;putrescine, 2.26 min; cadaverine, 2.43 min; heptanediamine, 2.83 min; spermidine, 3.42 min; and spermine, 4.41 min. With an additional column regeneration time of 3—4 min, the complete cycle per sample took less than 8 min at room temperture. Using a fluorescence detector, the lower limit of detection was less than 1 pmol per 6 μl injection volume. The fluorescence response was linear up to 200 pmol per 6 μl for each polyamine. The method is suitable for separation of polyamines from animal, plant and fungal sources.