Optimum Combination of Reversed-Phase Column Type and Mobile-Phase Composition for Gradient Elution Ion-Pair Chromatography of Amino Acids

Abstract

An optimum combination of reversed-phase column type and eluent composition was selected for the practical separation of amino acids by gradient elution ion-pair chromatography. A butyl-silica (C4) column with a concentration gradient elution between 5mM SDS-10mM HClO4 solution and acetonitrile provided the best resolution for amino acids. The C4 column was most adequate for reducing the chromatographic cycle time including the column regeneration. The use of 5mM SDS as ion-pairing agent in relatively low concentration is economical and practical for replicate and routine analyses. The developed HPLC system is applicable to the analysis of organic cations at low cost.