Massie, F.; MacKenzie, A.; Pandor, S.; Cremonini, M. A.; Moses, T.

Metabolomics is the comprehensive study of small molecules that provides a snapshot of an organism\'s physiological state. Reflecting phenotype more closely than genes or proteins, metabolites reveal changes linked to diseases, mutations, genetic interventions, and environmental stimuli. Recent technological advancements in metabolomics analysis through the application of ion mobility mass spectrometry have enhanced the comprehensive analysis of complex metabolic mixtures. However, pre-analytical bottlenecks in throughput and consistent extraction persist. We developed an automated, sample-agnostic metabolite extraction workflow for diverse liquid samples using an Agilent Bravo liquid handling platform. Here, we provide device, protocol, and form files for efficient sample processing to extract metabolites for global metabolomics analysis using liquid chromatography - mass spectrometry techniques.