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Probing the interaction mode in hydrophilic interaction chromatography
Umeå University, Faculty of Science and Technology, Department of Chemistry.
Merck SeQuant AB, Box 7956, S-90719 Umeå, Sweden.
Umeå University, Faculty of Science and Technology, Department of Chemistry.
2011 (English)In: Journal of Chromatography A, ISSN 0021-9673, E-ISSN 1873-3778, Vol. 1218, no 35, 5880-5891 p.Article in journal (Refereed) Published
Abstract [en]

This work aims at characterizing interactions between a select set of probes and 22 hydrophilic and polar commercial stationary phases, to develop an understanding of the relationship between the chemical properties of those phases and their interplay with the eluent and solutes in hydrophilic interaction chromatography. "Hydrophilic interaction" is a somewhat inexact term, and an attempt was therefore made to characterize the interactions involved in HILIC as hydrophilic, hydrophobic, electrostatic, hydrogen bonding, dipole-dipole, π-π interaction, and shape-selectivity. Each specific interaction was quantified from the separation factors of a pair of similar substances of which one had properties promoting the interaction mode being probed while the other did not. The effects of particle size and pore size of the phases on retention and selectivity were also studied. The phases investigated covered a wide range of surface functional groups including zwitterionic (sulfobetaine and phosphocholine), neutral (amide and hydroxyl), cationic (amine), and anionic (sulfonic acid and silanol). Principal component analysis of the data showed that partitioning was a dominating mechanism for uncharged solutes in HILIC. However, correlations between functional groups and interactions were also observed, which confirms that the HILIC retention mechanism is partly contributed by adsorption mechanisms involving electrostatic interaction and multipoint hydrogen bonding. Phases with smaller pore diameters yielded longer retention of solutes, but did not significantly change the column selectivities. The particle diameter had no significant effect, neither on retention, nor on the selectivities. An increased water content in the eluent reduced the multipoint hydrogen bonding interactions, while an increased electrolyte concentration lowered the selectivities of the tested columns and made their interaction patterns more similar.

Place, publisher, year, edition, pages
Elsevier , 2011. Vol. 1218, no 35, 5880-5891 p.
Keyword [en]
hydrophilic-interaction chromatography, retention factors, selectivity, column classification, partial least squares
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:umu:diva-46063DOI: 10.1016/j.chroma.2011.06.037PubMedID: 21803363OAI: oai:DiVA.org:umu-46063DiVA: diva2:436902
Available from: 2011-08-25 Created: 2011-08-25 Last updated: 2017-12-08Bibliographically approved
In thesis
1. Investigations of the retention mechanisms in hydrophilic interaction chromatography
Open this publication in new window or tab >>Investigations of the retention mechanisms in hydrophilic interaction chromatography
2013 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Hydrophilic interaction chromatography is well known as a powerful technique separation of polar and ionizable compound nowadays. However the retention mechanism of the technique is still under debate. Understanding retention mechanism would facilitate the method development using the technique and its future improvement. This was inspiring and became the goal of this thesis.

This work involves the characterization of the water enriched layer regarding to water and buffer salt accumulation. Twelve HILIC stationary phase with a diverse surface chemistry regarding to function groups and modification type were studied. Effect of water and salt on regarding to the retention mechanism was investigated by correlating the adsorption data to the retention of selected solutes

This also involved the characterization of interactions involve in the separation of 21 HILIC columns. Interactions was probe by retention ratio of pair solutes which are characteristic for each specific interaction. The data was evaluate using principle component analysis – a multivariable data analysis method. The model was comprehensive and its outcomes were confirmed by the studies on adsorptions of water and salts.

Place, publisher, year, edition, pages
Umeå: Umeå University, 2013. 42 p.
Keyword
HILIC, hydrophilic interaction chromatography, adsorption isotherm, water enriched layer, multivariate data analysis, retention mechanism, separation
National Category
Analytical Chemistry
Research subject
Analytical Chemistry
Identifiers
urn:nbn:se:umu:diva-68071 (URN)978-91-7459-630-4 (ISBN)
Public defence
2013-05-03, KBC Building, Room KB3A3, Umeå universitet, Umeå, 10:00 (English)
Opponent
Supervisors
Available from: 2013-04-12 Created: 2013-04-11 Last updated: 2013-05-14Bibliographically approved

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