Untargeted NMR Metabolomics of Serum

Metabolic biomarkers were quantified from serum samples using untargeted high-throughput proton nuclear magnetic resonance (NMR) spectroscopy metabolomics platform (Nightingale Health Plc, Helsinki, Finland). The details of the methodology used have been described previously (Soininen et al., 2015 (link)). The samples were barcoded for sample identification and kept frozen at −80°C for analysis. Metabolites were measured by a quantitative high-throughput NMR experimental set up for the simultaneous quantification of lipids and lipoprotein subclass profiling in 350 µL of serum. All liquid handling procedures were completed prior to the NMR studies, and the SampleJet robotic sample charger was set up at a cooled temperature to prevent sample deterioration. Every single metabolic measurement was subjected to a number of statistical quality control procedures and cross-referenced with a sizable collection of quantitative molecular data.