A high performance liquid chromatographic method coupled with an electrochemical detector for the separation and quantification of putrescine and cadaverine as their electroactive derivatives has been developed. Two different solvent column systems were used to determine putrescine, cadaverine, spermidine, spermine, and 1,3-diaminopropane (IS) in rat cerebral cortex. The lower detection limit was less than 80 fmol. The electrochemical response was linear in the range 80–240 ng/mL.