Advancing Metabolomic Characterization of Eukaryotic Pathogenic Systems: A Focus on High-Resolution Ion Mobility Separation of Lipid Isomers

Heidi M. Sabatini, Christopher D. Chouinard

Metabolomics provides a powerful lens for understanding eukaryotic pathogens by revealing small-molecule networks that drive infection, adaptation, and host interactions. These signatures can uncover virulence mechanisms, drug targets, and biomarkers not evident from genomic or proteomic data alone. Despite its standing as the “gold-standard” in metabolomics, LC-MS-based methods can still struggle with separation and identification of novel metabolites in complex mixtures, especially for challenging isomers. Our work has recently focused on high-resolution ion mobility (HRIM) as a complement to existing LC-MS workflows, as the technique nests nicely and provides an additional dimension of separation based on ion size, shape, and charge. Herein we apply several HRIM approaches aimed at resolving challenging lipid isomers relevant to eukaryotic pathogen systems.