Simultaneous separation, identification, and quantification of lipid isomers using non-covalent chemistry-based enhancement of tandem ION mobility-mass spectrometry

The National Institute of General Medical Sciences, part of the Department of Health and Human Services National Institutes of Health, awarded Brigham Young University $406,576 on September 1, 2026, under the Biomedical Research and Research Training program (CFDA 93.859) to develop an analytical platform for simultaneous separation, identification, and quantification of lipid isomers using non-covalent chemistry-based enhancement of tandem ion mobility-mass spectrometry.

The research integrates non-covalent chemical modification strategies with cyclic ion mobility spectrometry coupled to tandem mass spectrometry to enable gas-phase separation of lipid isomers with enhanced resolution. The work addresses the current lack of analytical technologies with sufficient resolution and throughput to distinguish and quantify lipid isomers in complex biological samples, despite evidence that structural variations in isomers—such as double-bond position, stereochemistry, or glycosidic linkage—profoundly influence biological activity and disease relevance. Examples include glycosphingolipid isomers glucosylceramide and galactosylceramide associated with Gaucher and Krabbe diseases, respectively, and positional isomers of unsaturated lipids linked to breast cancer. The research team has demonstrated that non-covalent modifications enable isomer separation, identification, and relative quantification in small molecule systems and is extending these methods to biologically relevant lipid mixtures, including glycosphingolipid stereoisomers. Recent implementation of gas-phase ion/ion reactions within the platform further expands the chemical space accessible for isomer discrimination.

Work is performed in Provo, Utah. The period of performance runs through May 31, 2031.

Abstract

Lipids are central to human physiology, functioning in membrane structure, energy storage, and cellular signaling. Many biologically active lipids exist as isomers-molecules that share the same chemical formula but differ in the arrangement of atoms or bonds. These structural variations, such as double-bond position, stereochemistry, or glycosidic linkage, can profoundly influence biological activity and disease relevance. For example, the glycosphingolipid isomers glucosylceramide and galactosylceramide are associated with gaucher and krabbe diseases, respectively, while positional isomers of unsaturated lipids have been linked to breast cancer. Despite growing evidence for isomer-specific lipid functions, current analytical technologies lack the resolution and throughput needed to routinely distinguish and quantify these species in complex biological samples. Our laboratory is developing a unified analytical platform that integrates non-covalent chemical modification strategies with high-resolution ION mobility spectrometry (IMS), specifically cyclic IMS (cims), coupled to tandem mass spectrometry (MS). This approach enables gas-phase separation of lipid isomers with enhanced resolution, compatibility with multiple sample types, and adaptability across lipid classes. We have demonstrated that non-covalent modifications enable isomer separation, identification, and relative quantification in small molecule systems, and are now extending these methods to biologically relevant lipid mixtures, including glycosphingolipid stereoisomers. Recent implementation of gas-phase ION/ION reactions within our cims platform further expands the chemical space accessible for isomer discrimination. Over the next five years, our research will focus on optimizing non-covalent modification chemistries, validating their performance across diverse lipid classes, and applying these tools to total lipid extracts. This work will establish a broadly applicable lipidomics platform capable of resolving isomeric complexity, enabling the discovery of novel lipid biomarkers and elucidation of isomer-specific metabolic and inflammatory pathways. By advancing the analytical capabilities of lipidomics, this research supports NIH priorities in biomarker discovery and mechanistic understanding of disease.