Automated Quantitation of Polyamines by Improved Cation-Exchange High-Performance Liquid Chromatography Using a Pump Equipped With a Plunger Washing System

Abstract

Abstract An improved method for simple, precise, and sensitive quantitation of polyamines[putrescine, spermidine, spermine, cadaver-ine(in part) and 1, 6-diaminohexane] by cation-exchange high performance liquid chromatography is described. Postcolumn deriv-atization using the o-phthalaldehyde method was employed to detect them. Analysis took 45 minutes and the minimum detectable amount of each polyamine was 10 – 20 pmol/μ1. An isocratic pump equipped with a plunger washing system and a mobile phase of pH 5.3 are the advantages of this improved method. Consequently, an operator can carry out routine and automated polyamine analysis without any hesitation regarding maintenance of the pump or column damage due to crystal precipitation from the mobile phase on or in the pump, which has been a problem previously encountered with cation-exchange resin. Reproducibility of the day to day precision and duplicate determination, and simultaneous reproducibility of the standard mixture were calculated (CV<2.2%).