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Molecular Enzymology and Drug Targets

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Abstract

Modern Developments and Emerging Directions in Hydrophilic Interaction Chromatography-Mass Spectrometry for Metabolomics and Proteomics

Shivanshi Singh*

Proteomics and metabolomics have drawn more interest during the past ten years than any other -omics methods. Both methods have matured to the point where they are relevant in many clinical applications, including as the discovery of biomarkers, enhanced illness diagnosis, staging, and prognosis, as well as a greater understanding of many physiological processes. Due to its simplicity and reproducibility, reversed-phase liquid chromatography mass spectrometry is regarded analytically as the gold standard in proteomics and metabolomics. The complexity of the proteome cannot be resolved by RPLC-MS alone, because highly polar metabolites are often poorly preserved. Due to its orthogonal separation process, hydrophilic interaction chromatography in this context constitutes an appealing supplementary technique. This review provides a summary of the research literature regarding the use of HILIC-MS for proteomics and metabolomics. In contrast to proteomics, which discusses the analysis of complex samples and protein post-translational modifications therein using bottom-up, middle-up/down proteomics, and intact protein analysis, metabolomics focuses on the analysis of bioactive lipids, amino acids, organic acids, and nucleotides/nucleosides. The technical aspects of HILIC-MS utilisation in proteomics and metabolomics are covered in the review, with special focus paid to the stationary phases, mobile phase parameters, injection volume, and column temperature. Additionally shown and discussed are recent trends and advancements in the use of HILIC-MS in proteomics and metabolomics, emphasising the benefits the method can offer in addition to or as a complement to RPLC-MS as well as the current drawbacks and potential remedies.

Keywords

Electrostatic repulsion hydrophilic; interaction chromatography; Hydrophilic interaction chromatography; HILIC-MS; Metabolomics; Proteomics

Published Date: 2023-04-28; Received Date: 2023-04-03