Metabolomics Analysis of Urinary Extracellular Vesicles by Nuclear Magnetic Resonance and Liquid Chromatography–Mass Spectrometry

Extracellular vesicle (EV) release and their content are influenced by diverse clinical conditions. EVs participate in inter-cellular communication and have been postulated as reflectors of the pathophysiology of the cells, tissues, organs or the whole system with which they are in contact. Urinary EVs have been proved to reflect pathophysiology not only of renal system related diseases constituting an additional source of potential biomarkers easily accessible in a non-invasive way. The interest in EVs cargo has been mostly focused on proteinsProteins and nucleic acids and more recently it has been extended to metabolitesMetabolites. MetabolitesMetabolites represent the downstream changes in the genome, transcriptome, and proteome as a reflection of processes occurring in living organisms. For their study, nuclear magnetic resonance (NMR)Nuclear magnetic resonance (NMR)Nuclear magnetic resonance (NMR) and mass spectrometryMass spectrometry in tandem (LC-MS/MS) are widely used. NMRNuclear magnetic resonance (NMR) is a reproducible and non-destructive technique and we show here methodological protocols for the metabolomicsMetabolomics analysis of urinary EVs by NMRNuclear magnetic resonance (NMR). Additionally, we also describe the workflow for a targeted LC-MS/MS analysis that is extensible to untargeted studies.