Rapid chromatography combined with trapped ion mobility mass spectrometry for analysis of derivatized small molecules

ABSTRACT: Liquid chromatography (LC) separates biomolecules based on their physicochemical properties. Reproducible retention times support compound identification, while efficient separation of structurally similar compounds is essential for accurate quantification. However, conventional LC methods are often time-intensive, limiting analytical throughput in large-scale and clinical applications. Here, we present rapid LC–trapped ion mobility spectrometry–tandem mass spectrometry (LC–TIMS–MS/MS) approaches that substantially reduce analysis time while preserving chromatographic resolution and quantitative reliability, with run times of around 2 minutes or below. Accelerated separations were achieved using shorter columns and elevated flow rates coupled to a trapped ion mobility mass spectrometer. The added ion mobility dimension improves molecular specificity and enhances confidence in compound annotation. To maximise metabolome coverage in a single chromatographic analysis, and to further increase throughput, we implemented automated derivatization of carboxylic acids, ketones, aldehydes, and amino acids using a liquid-handling platform, minimizing rate-limiting steps in sample preparation. The resulting workflow is well suited for high-throughput small-molecule analysis in large cohort studies and screening applications. The method was applied to the analysis of short-chain fatty acids, tricarboxylic acid cycle intermediates, and amino acids in human plasma and fecal samples.

INSTRUMENT(S): Liquid Chromatography MS - negative - reverse-phase, Liquid Chromatography MS - positive - reverse-phase

PROVIDER: MTBLS14813 | MetaboLights | 2026-06-30

REPOSITORIES: MetaboLights