Determination of polyamines in human blood by electron-capture gas-liquid chromatography

Abstract

The present work was undertaken to develop a sensitive and selective method for the estimation of putrescine, spermidine and spermine in human blood employing electron-capture gas--liquid chromatography. Polyamines were derivatized with heptafluorobutyric anhydride. The heptafluorobutyric derivatives of polyamines could be well resolved within 15 min under a temperature programme. The detection limit was 0.1 pmol for putrescine and cadaverine, and 0.02 pmol for spermidine and spermine. The method was applied to polyamine determinations in erythrocytes from human blood. For pre-separation of the polyamines from other compounds, a simple clean-up method utilizing an activated Permutit has been devised. Major interfering substances could be removed by the batchwise Permutit treatment. The mean values of spermidine and spermine concentrations, and the spermidine/spermine ratio in erythrocytes obtained from normal subjects (n = 11) were similar to reported values. The analytical procedure is thought to be applicable to various biological materials.