LC-MS Metabolite Profiling Workflow

LC-MS analysis was performed on a quadrupole-time-of-flight (Q-TOF) 6550 mass spectrometer (Aglient Technologies, Santa Clara, CA) with an electrospray ionization source. The mass spectrometer was interfaced with an Agilent 1290 Infinity II UPLC system (Aglient Technologies, Santa Clara, CA). Metabolites were analyzed in the positive mode over a m/z range of 50–1000 with a C18 T3 reverse-phased column (Waters Corporation, Milford, MA). MS/MS data were generated on the Q-TOF for the identification of perturbed metabolites. The XCMS Online sever (https://xcmsonline.scripps.edu, version 3.5.1) was applied for peak picking, alignment, integration, and extraction of the peak intensities. A two-tailed Welch’s t-test was used for the assessment of metabolite differences between control and treatment groups. The programs of MS-DIAL^{56 (link)} (version 2.70) and MS-FINDER^{57 (link)} (version 2.20) were used for the identification of metabolites.