Microdetermination of Polyamines (Putrescine, Cadaverine, Spermidine, And Spermine) Using Polyamide Thin-Layer

Abstract

A fluorometric method for the determination of polyamines (putrescine, cadaverine, spermidine, and spermine) using polyamide thin-layer is descsibed. Dansylated polyamines obtained after the reaction with dansyl-chloride at pH 9.5 at 40°C for 1 h were extracted into toluene. An aliquot of the extract was spotted on the polyamide thin-layer plate, and developed with a solvent system of water-formic acidtriethylamine (54 : 15 : 2). Spots were measured with a scanning fluorometer. The fluorescence intensity of dansylated polyamines was stable on the plate at least 24 h. The relative standard deviation of the fluorescence intensity obtained on the same thin-layer plate was (1. 5~5. 0) % at 200 pmol/spot, and the lowest limit of determination was 10 pmol/spot for bis-DNSputrescine, bis-DNS-cadaverine, and tri-DNS-spermidine, and 5 pmole/spot for tetra-DNS-spermine. This method was successfully applied for the determination of polyamines in tissues of male rats and guinea pigs. © 1982, The Japan Society for Analytical Chemistry. All rights reserved.