Improved analysis for urinary polyamines by use of high-voltage electrophoresis on paper

This method for assay of polyamines (putrescine, spermidine, and spermine) in large numbers of urine samples is based on preliminary analyte isolation by use of a Dowex 50W column, separation by high-voltage paper electrophoresis, reaction with ninhydrin to detect the separated amines, and subsequent direct assay with a dual-wave-length thin-layer chromatography scanner by the zigzag scanning method. By optimizing the conditions for the Dowex 50W column and high-voltage paper electrophoresis, putrescine, cadaverine, spermidine, and spermine were completely separated. The major technical improvements are room-temperature collection of urine with toluene, shorter hydrolysis time, complete separation of putrescine from cadaverine and histamine, and elimination of interference with spermine measurement. Polyamine concentrations as measured by this method agreed with those obtained by use of an amino acid analyzer. Reference intervals by this method were similar to those reported by Russell (Clin. Chem. 23: 22, 1977) with an automated amino acid analyzer. Putrescine, spermidine, and spermine excretion in urine from patients with blood cancers and solid cancers was significantly increased; information on concentrations of putrescine and spermidine were especially useful for following the clinical efficacy of cancer treatments.