Related Experiment Video
Updated: Jul 17, 2026

Synthesis and Purification of Iodoaziridines Involving Quantitative Selection of the Optimal Stationary Phase for Chromatography
Published on: May 16, 2014
Behaviour of sulphonated azodyes in ion-pairing reversed-phase high-performance liquid chromatography
Dana Vanerková1, Pavel Jandera, Jaroslav Hrabica
1Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Nám. Cs. Legií 565, CZ-532 10 Pardubice, Czech Republic.
Abstract:
Ion-pairing reversed-phase high-performance liquid chromatography (RP-HPLC) operation conditions were studied to obtain information useful for development and optimization of separation methods suitable for HPLC/MS analysis of dyes. The retention of eight sulphonated azodyes with widely differing structures (1-5 acid groups, molecular weight range 350-1220) was measured in mobile phases containing various ion-pairing reagents in aqueous-methanolic mobile phases. The effects of the type and of the concentration of ammonium acetate, tetrabutylammonium hydrogen sulphate and five di- and tri-alkylammonium acetate ion-pairing reagents on the chromatographic behaviour of dyes were compared in mobile phases with varying concentrations of methanol. Structural effects on the retention of dyes were studied in detail. The retention scale based on lipophilic and polar indices can be used for optimization of mobile phase for HPLC/MS of dyes and, on the other hand, may provide some information on the structure of unknown dyes.
Related Concept Videos
Ion-Exchange Chromatography
High-Performance Liquid Chromatography: Introduction
In HPLC, two phases play a critical role in the separation process:
Ion Exchange
Aryldiazonium Salts to Azo Dyes: Diazo Coupling
High-Performance Liquid Chromatography: Elution Process
Silica Gel Column Chromatography: Overview
Polar components tend to bind strongly to the silica gel, causing them to move slowly through the column. In contrast, nonpolar compounds...

