Managing sample introduction problems in hydrophilic interaction liquid chromatography

Taylor, Mark R.; Kawakami, Jane; McCalley, David V.

Abstract

Sample injection can cause serious problems in hydrophilic interaction liquid chromatography (HILIC) when the injection solvent has higher elution strength than the mobile phase (mp). It can lead to asymmetric peak shapes and poor efficiency. The problem can occur when the mp contains a high proportion of organic e.g. 95% acetonitrile (a weak solvent) whereas the injection solvent contains a higher proportion of water (a strong solvent) that is necessary to dissolve polar samples. We investigated different strategies to overcome this problem. A simple method is pre-column dilution where the injector is programmed to deliver a plug of weak solvent (e.g. pure acetonitrile) along with the sample dissolved in a solvent with higher water content than the mp. Another option is to use alternative organic solvents to acetonitrile in the injection solvent, e.g. isopropanol, acetone or tetrahydrofuran, that may give enhanced sample solubility. The role of the volume of injection solvents was investigated as well as the possible effects of mass overload on the results. The use of small sample volumes is always recommended to reduce mismatch effects.

• Journal Article Type: Article
• Acceptance Date: Apr 19, 2023
• Online Publication Date: Apr 22, 2023
• Publication Date: Jul 5, 2023
• Deposit Date: May 15, 2023
• Publicly Available Date: Apr 23, 2025
• Journal: Journal of Chromatography A
• Print ISSN: 0021-9673
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 1700
• Article Number: 464006
• DOI: https://doi.org/10.1016/j.chroma.2023.464006
• Keywords: HPLC; HILIC; sample injection
• Public URL: https://uwe-repository.worktribe.com/output/10723628
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S0021967323002327?via%3Dihub