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Water uptake on polar stationary phases under conditions for hydrophilic interaction chromatography and its relation to solute retention
Umeå University, Faculty of Science and Technology, Department of Chemistry. Merck SeQuant AB, Umeå. (Knut Irgum)
Merck SeQuant AB, Umeå.
Umeå University, Faculty of Science and Technology, Department of Chemistry.
2013 (English)In: Journal of Chromatography A, ISSN 0021-9673, E-ISSN 1873-3778, Vol. 1320, 33-47 p.Article in journal (Other academic) Published
Abstract [en]

In hydrophilic interaction chromatography, water is known to accumulate on the stationary phase to form a water enriched layer, which is believed to play an important role in the retention mechanism. To gain a better understanding retention mechanism in HILIC, we have determined the water uptake on twelve different HILIC stationary phases. Non-modified and monomerically functionalized silica phases followed a pattern of monolayer formation followed by multiple layer adsorption, while the water uptake on polymerically functionalized silica stationary phase showed the characteristics of formation and swelling of hydrogels. This difference in the nature of water accumulation was found to be related to different water uptake patterns when methanol and tetrahydrofuran were added to 80:20 % (v/v) acetonitrile/water by replacing 5 % of the acetonitrile as tertiary solvents, and also when ammonium acetate was added as buffering electrolyte. The relationship between water uptake and retention mechanism was investigated by looking at the correlation between retention factors of neutral analytes and phase ratios of HILIC columns, calculated either as surface area (adsorption) or volume of the water layer enriched from the acetonitrile/water eluent (partitioning). Regardless of the adsorption or partitioning mechanism, the interaction of neutral analytes and stationary phase could be mainly the hydrogen bonding between analytes and the accumulated water in the water enriched layer.

Place, publisher, year, edition, pages
Elsevier, 2013. Vol. 1320, 33-47 p.
Keyword [en]
HILIC, HPLC, Retention mechanism, Stationary phases, Eluents
National Category
Analytical Chemistry Biochemistry and Molecular Biology
URN: urn:nbn:se:umu:diva-68067DOI: 10.1016/j.chroma.2013.09.061ISI: 000327922100005OAI: diva2:615721
Swedish Research Council, 2008-3621, 2012-4000

Manuscript version included in dissertation with the title: Water uptake on polar stationary phases under HILIC conditions and its relation to solute retention

Available from: 2013-04-11 Created: 2013-04-11 Last updated: 2014-02-25Bibliographically 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.
HILIC, hydrophilic interaction chromatography, adsorption isotherm, water enriched layer, multivariate data analysis, retention mechanism, separation
National Category
Analytical Chemistry
Research subject
Analytical Chemistry
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)
Available from: 2013-04-12 Created: 2013-04-11 Last updated: 2013-05-14Bibliographically approved

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Dinh, Ngoc PhuocIrgum, Knut
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