Metabolic extracts were analyzed four times using hydrophilic liquid chromatography (HILIC) and reverse phase liquid chromatography (RPLC) separation in both positive and negative ionization modes as previously described124 (link). Data were acquired on a Thermo Q Exactive plus mass spectrometer equipped with a HESI-II probe and operated in full MS scan mode. MS/MS data were acquired on pool samples consisting of an equimolar mixture of all the samples in the study. HILIC experiments were performed using a ZIC-HILIC column 2.1×100 mm, 3.5μm, 200Å (Merck Millipore) and mobile phase solvents consisting of 10mM ammonium acetate in 50/50 acetonitrile/water (A) and 10 mM ammonium acetate in 95/5 acetonitrile/water (B). RPLC experiments were performed using a Zorbax SBaq column 2.1 × 50 mm, 1.7 μm, 100Å (Agilent Technologies) and mobile phase solvents consisting of 0.06% acetic acid in water (A) and 0.06% acetic acid in methanol (B). Data quality was ensured by (i) injecting 6 and 12-pool samples to equilibrate the LC-MS system prior to run the sequence for RPLC and HILIC, respectively, (ii) sample randomization for metabolite extraction and data acquisition, and (iii) checking mass accuracy, retention time and peak shape of internal standards in every samples.