Urine polyamines determination using dansyl chloride derivatization in solid-phase extraction cartridges and HPLC

The derivatization of biogenic amines such as putrescine, cadaverine, sperm idine and spermine with dansyl chloride in solid phase extraction cartridge s is described. Different types of filling materials were tested in order t o have the highest retention of the different analytes. The best results we re obtained by using C18 cartridges. The optimal conditions were: amine sol ution buffered at pH 12, 2 mM dansyl chloride (acetone-bicarbonate solution 20 mM (pH 9-9.5), 2 + 3 v/v) as reagent concentration, room temperature an d 30 min reaction time. The developed procedure was applied to the determin ation of these polyamines in urine samples from healthy controls and cancer patients using HPLC with 1,7-diaminoheptane as internal standard. The conc entrations ranged from 0.5 to 5 mu g mL(-1) and the detection limits were 1 0 ng mL(-1) for all polyamines. By concentrating the urine extracts, the de tection limits were improved down to 2 ng mL(-1). The accuracy and the prec ision of the method were tested. The proposed dansylation method is advanta geous with respect to solution dansylation. It improves the total analysis time, avoids high temperatures that can affect the thermal stability of the derivatives and could make possible the automation of the procedure.