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Polymer-based monolithic microcolumns for hydrophobic interaction chromatography of proteins.
Umeå University, Faculty of Science and Technology, Chemistry.
Umeå University, Faculty of Science and Technology, Chemistry.
Umeå University, Faculty of Science and Technology, Chemistry.
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2006 (English)In: Journal of Separation Science, ISSN 1615-9306, Vol. 29, no 1, 25-32 p.Article in journal (Refereed) Published
Abstract [en]

Monolithic capillary columns for hydrophobic interaction chromatography (HIC) have been prepared by thermally initiated, single-step in situ polymerization of mixtures of monovinyl monomers including butyl methacrylate and/or 2-hydroxyethyl methacrylate, with a divinyl crosslinker glycerol dimethacrylate or 1,4-butanediol dimethacrylate using two different porogen systems. Two porogenic solvent mixtures were used; one "hydrophilic", consisting of water, butanediol, and propanol, and one "hydrophobic," comprising dodecanol and cyclohexanol. The porous structures of the monoliths were characterized and their performance was demonstrated with a separation of a mixture of myoglobin, ribonuclease A, and lysozyme under conditions typical of HIC.

Place, publisher, year, edition, pages
2006. Vol. 29, no 1, 25-32 p.
Keyword [en]
Capillary, Column, HIC, Hydrophobic interaction chromatography, Monolith, Protein separation
URN: urn:nbn:se:umu:diva-11762DOI: doi:10.1002/jssc.200500239OAI: diva2:151433
Available from: 2007-04-23 Created: 2007-04-23 Last updated: 2009-09-17Bibliographically approved
In thesis
1. Hydrophilic Separation Materials for Liquid Chromatography
Open this publication in new window or tab >>Hydrophilic Separation Materials for Liquid Chromatography
2007 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The main focus of this thesis is on hydrophilic interaction chromatography (HILIC) and the preparation of stationary phases for HILIC. The mechanism of HILIC is also discussed; a large part of the discussion has been adapted from a review written by me and professor Irgum for the Journal of Separation Science (ref 34). By reevaluating the literature we have revealed that the notion of HILIC as simply partitioning chromatography needed modification. However, our interest in the HILIC mechanism was mainly inspired by the need to understand how to construct the optimal HILIC stationary phase. The ultimate stationary phase for HILIC is still not found. My theory is that a non-charged stationary phase capable of retaining a full hydration layer even at extreme acetonitrile (> 85%) concentrations should give a HILIC stationary phase with a more pure partitioning retention behavior similar to that of a swollen C18 reversed phase. The preparation of a sorbitol methacrylate grafted silica stationary phase is one of our attempts at producing such a stationary phase. The preparation of such a grafted silica has been performed, but with huge difficulty and this work is still far from producing a column of commercial quality and reprodicibility.

This thesis also discusses a new method for the initiation of atom transfer radical polymerization from chlorinated silica. This new grafting scheme theoretically results in a silica particle grafted with equally long polymer chains, anchored to the silica carrier by a hydrolytically stable silicon-carbon bond. The hydrolytic stability is especially important for HILIC stationary phases due to the high water concentration at the surface.

Place, publisher, year, edition, pages
Umeå: Kemi, 2007. 24 p.
Chromatography, HILIC, Hydrophilic, Stationary phase
National Category
Chemical Sciences
urn:nbn:se:umu:diva-1350 (URN)978-91-7264-406-9 (ISBN)
Public defence
2007-10-05, KB3A9, KBC-Huset, Umeå Universitet, Umeå, 10:00 (English)
Available from: 2007-09-14 Created: 2007-09-14 Last updated: 2009-09-17Bibliographically approved

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