Managing the column equilibration time in hydrophilic interaction chromatography

McCalley, David V.

Abstract

For a wide variety of hydrophilic interaction chromatography stationary phases, a repeatable partial equilibration was demonstrated in gradient elution after purging with as little as 12 column volumes of mobile phase. Relative standard deviations of retention time of on average ~0.15% could be obtained after 1 or 2 conditioning (blank) runs. The equilibration period must be kept strictly constant, otherwise selectivity changes occur, but this is not problematic on modern instruments. Partial equilibration was largely independent of stationary phase or gradient slope. Alternatively, full column equilibration is favoured for stationary phases that do not trap extensive water layers, and for materials with a wider pore size that have a lower surface area. Temperatures somewhat above ambient also shorten the equilibration time. Some stationary phases under optimum conditions can achieve full column equilibration using purging with ~12 column volumes, which is useful for rapid set-up of isocratic separations or for conventional gradient analysis.

• Journal Article Type: Article
• Acceptance Date: Oct 23, 2019
• Online Publication Date: Oct 24, 2019
• Publication Date: Feb 8, 2020
• Deposit Date: Oct 25, 2019
• Publicly Available Date: Oct 25, 2020
• Journal: Journal of Chromatography A
• Print ISSN: 0021-9673
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 1612
• Article Number: 460655
• DOI: https://doi.org/10.1016/j.chroma.2019.460655
• Keywords: Organic Chemistry; Analytical Chemistry; Biochemistry; General Medicine
• Public URL: https://uwe-repository.worktribe.com/output/4103795