HPLC determination of polyamines in the femtomole range

A new method for the chromatographic determination of polyamines is presented. The procedure consists of the reaction of 9-fluorenylmethyl chloroformate (FMOC-Cl) with polyamines at 50 +/- 1 degrees C for 10 min, followed by stepwise elution chromatography. The reaction is simple and the derivatives are stable. All the polyamine-FMOC derivatives can be separated within 13 min. The linearity of this method is satisfactory in the range of 0.5-100 pmol. The detection limits for putrescine, cadaverine, spermidine and spermine are 83.3 fmol, 109 fmol, 25.5 fmol and 22.7 fmol respectively.