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The Effects of the Column Length on the Efficiency of Capillary Zwitterionic Organic Polymer Monolithic Columns in HILIC Chromatography

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Abstract

Organic polymer monolithic columns of different lengths have been prepared in 320 µm i.d. fused silica capillary by in situ radical polymerization of N,N-dimethyl-N-methacryloxyethyl-N-(3-sulfopropyl) ammonium betaine as a zwitterionic functional monomer and bisphenol A glycerolate dimethacrylate as a crosslinking monomer in the presence of porogenic solvents. The zwitterionic monolithic columns are intended for separations of polar compounds in hydrophilic interaction chromatography (HILIC). The effects of the capillary column length, from 115 to 175 mm, on separation efficiency, were investigated under HILIC conditions, using 95:5 acetonitrile in water as the mobile phase. The extra-column contributions to band broadening significantly decrease the efficiency (apparent height equivalent to a theoretical plate), especially for weakly retained samples, and increase with diminishing column length. The experimental height equivalents of theoretical plate, HETP, were corrected for the extra-column contributions, which were determined for a series of columns by extrapolation to zero column length. On a 175 mm long column, the column efficiency, HETP = 16.5 μm, measured at the optimum linear flow velocity of 0.5 mm s−1, improved to HETP = 5 µm, after correction for extra-column contributions. For more strongly retained small polar compounds, interactions with zwitterionic groups and (or) water adsorbed inside the pores decrease the column efficiency at higher flow rates.

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Acknowledgments

This work was funded by the Czech Science Foundation (the Grant Agency of the Czech Republic) under the project No. 206/12/0398.

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Authors and Affiliations

  1. Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentska 573, 53210, Pardubice, Czech Republic

    Pavel Jandera & Magda Staňková

Authors
  1. Pavel Jandera
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  2. Magda Staňková
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Correspondence to Pavel Jandera.

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Published in the topical collection 20th International Symposium on Separation Sciences in Prague with guest editors Aleš Horna and Pavel Jandera.

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Jandera, P., Staňková, M. The Effects of the Column Length on the Efficiency of Capillary Zwitterionic Organic Polymer Monolithic Columns in HILIC Chromatography. Chromatographia 78, 853–859 (2015). https://doi.org/10.1007/s10337-015-2848-4

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  • DOI: https://doi.org/10.1007/s10337-015-2848-4

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