Laboratory methods
Untargeted metabolite profiling was performed by Metabolon (Munich, Germany) using a Waters ACQUITY ultrahigh performance LC (UPLC; Elstree, UK) and Thermo Scientific Q-Exactive high-resolution MS with a heated electrospray ionisation (HESI-II) source (Waltham, USA). Two aliquots were analysed using acidic positive ion conditions, which included optimisation step for accurate determination of hydrophilic or hydrophobic compounds. Two further aliquots were analysed using negative ionisation mode. The MS analysis alternated between full scan MS (scan range 70–1000 m/z, scan rate 4 Hz) and data-dependent MSCompounds were identified using retention index, mass match ±10 ppm, and MS/MS forward and reverse scores between the sample data and the authenticated standards using a library, maintained by Metabolon. Peaks were quantified using AUC; raw data were normalised to correct for instrument inter-day tuning differences (median 1.0 for each run).Lipids were analysed using the Metabolon complex lipid panel. Lipids were extracted using a modified Bligh-Dyer extraction (methanol/water/dichloromethane) with 