In-Depth Performance Analysis and Comparison of Monolithic and Particulate Zwitterionic Hydrophilic Interaction Liquid Chromatography Polymer Columns.

ABSTRACT

The kinetic performance of different zwitterionic hydrophilic interaction liquid chromatography polymer columns is evaluated and compared in-depth. For this purpose, two lab-made monolithic columns, synthesized with different crosslinkers, and a commercial particle packed column are considered. It is found that performance evaluation techniques, such as comparing plate height curves or fitted A-, B- and C-terms, obtained by fitting experimental plate height data to a plate height model, are complicated by the determination of a reliable characteristic length. This is due to the very different morphology of these column types, and the heterogeneity of the monolithic columns. The occurrence of a convective flow through the packed particle column further complicates the interpretation of the obtained fitting parameters, as part of the C-term is wrongfully attributed to the A-term. Therefore, the use of the kinetic plot method is suggested for the comparative evaluation of these columns, as kinetic plots do not require the determination of a characteristic length, nor rely on any fitting parameters. With the kinetic plot method, it is demonstrated that the lab-made monolithic columns outperform the packed particle column for plate counts between 10,000 and 800,000. This is attributed to the higher column efficiency of these columns, due to their small domain and skeleton sizes, and their high permeability, resulting from their high external porosity and the occasional occurrence of preferential flow paths.