Ion-exchange chromatographic separation and fluorometric detection of urinary polyamines.

An ion-exchange chromatographic procedure has been developed for the separation and fluorometric detection of the urinary polyamines 1,3-diaminopropane, putrescine, spermidine, cadaverine, and spermine. The polyamines were separated on a 15 x0.45 cm cation-exchange column (70”) employing a combined pH-salt gradient. The separation conditions were designed to enable utilization of the sensitive reagent fluorescamine as a means of monitoring these amines in physiologic fluids. This non-fluorescent reagent forms a fluorophor with most primary amines. Experiments were carried out to determine the effects of several variables such as pH, reagent concentration, and flow-rate. The method was successfully applied to both normal and pathologic samples of human urine.