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Some factors that can lead to poor peak shape in hydrophilic interaction chromatography, and possibilities for their remediation

Heaton, James C.; Heaton, James; McCalley, David V.

Authors

James C. Heaton

James Heaton



Abstract

© 2015 Elsevier B.V. Some factors which present difficulties for obtaining good peak shape in hydrophilic interaction chromatography (HILIC) were studied. The effect of injection solvent composition and volume was systematically investigated using a selection of weak and stronger basic compounds on a hybrid bare silica phase. Increasing the mismatch between the injection solvent (range 95-0% ACN v/v) and the mobile phase (maintained at 95% ACN v/v) gave increasing deterioration in peak shape. With the 2.1 mm ID columns used, injections in the mobile phase of increasing volume (1-20 μL) gave poorer peak shape, but the magnitude of the effect was considerably smaller than that of solvent mismatch over this range. Some solute structural features such as galloyl (trihydroxy benzene), catechol (benzene diol) and phosphate (in nucleotides) gave serious peak tailing, attributed to interactions with metals in the stationary phase or the chromatographic hardware. These undesirable effects can be moderated by including complexing agents in the mobile phase, by changing the stationary phase chemistry, or by altering the mobile phase pH.

Citation

Heaton, J. C., Heaton, J., & McCalley, D. V. (2016). Some factors that can lead to poor peak shape in hydrophilic interaction chromatography, and possibilities for their remediation. Journal of Chromatography A, 1427, 37-44. https://doi.org/10.1016/j.chroma.2015.10.056

Journal Article Type Article
Acceptance Date Oct 16, 2015
Publication Date Jan 4, 2016
Journal Journal of Chromatography A
Print ISSN 0021-9673
Electronic ISSN 1873-3778
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 1427
Pages 37-44
DOI https://doi.org/10.1016/j.chroma.2015.10.056
Keywords HILIC, peak shape, catecholamines, acids, nucleotides
Public URL https://uwe-repository.worktribe.com/output/915530
Publisher URL http://dx.doi.org/10.1016/j.chroma.2015.10.056

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