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The role of HILIC in uni- and two-dimensional separations of pharmaceuticals, supplements, and low-molecular metabolites

Authors: Česla Petr | Kubát Miroslav | Tošovská Barbora | Kašpar Michal

Year: 2024

Type of publication: ostatní - přednáška nebo poster

Page from-to: nestránkováno

Titles:

Language	Name	Abstract	Keywords
eng	The role of HILIC in uni- and two-dimensional separations of pharmaceuticals, supplements, and low-molecular metabolites	Hydrophilic interaction liquid chromatography (HILIC) is an alternative to reversed-phase liquid chromatography (RP-LC) for separation of highly polar ionic or ionizable analytes, with orthogonal separation mechanims to RP-LC, and straighforward coupling to mass spectrometric detection. A wide range of stationary phases is currently commercially available for HILIC separations ranges from bare silica over various chemical moieties bonded on silica, hybrid silica, or polymeric support Most HILIC columns provide a dual retention mechanism combining RP and HILIC depending on the composition of the mobile phase, which can be beneficial especially in designing two-dimensional separation systems. In this work, a novel approach to the selection of a suitable type of stationary phase for target application was proposed and tested using several mixtures of polar compounds including nucleosides, phenones, oligosaccharides, and low-molecular glutathione metabolites and thiols related to glutathione metabolism. The glutathione-related metabolites are important indicators of cellular oxidative stress and biomarkers of many diseases, e.g., cataracts, cancer, neurodegenerative disease, pneumonia, and cystic fibrosis. The HILIC separation coupled with mass spectrometry was used for the assessment of glutathione metabolism without the need for derivatization, which was demonstrated on the analyses of cell lysates in the metabolic studies of glutathione metabolism. In HILIC, water can be partially or entirely replaced by another polar solvent, usually an alcohol such as methanol, ethanol, propan-2-ol. Thus the possibility of modifying the retention and selectivity of separation of polar analytes was tested employing alternative polar modifier to acetonitrile, and the retention model was established and tested for optimization of gradient separations with ternary mobile phases.