Metabolomic Profiling

The collected intestinal tissue samples for metabonomic analysis. The metabolites were extracted with 120 μL of precooled 50% methanol, vortexed mixed for 1 min, and incubated at room temperature for 10 min. After centrifugation at 4,000 g for 20 min, the supernatants were transferred into new 96 well plates. The samples were stored at -80 °C before the LC-MS analysis. First, all chromatographic separations were performed with an ultra-performance liquid chromatography (UPLC) system (SCIEX, UK). An ACQUITY UPLC T3 column (100 mm * 2.1 mm, 1.8 µm, Waters, UK) was used for the reversed phase separation. A high-resolution TripleTOF 5600plus tandem mass spectrometer (SCIEX, UK) was used to detect metabolites eluted from the column. The Q-TOF was operated in both positive and negative ion modes. LC-MS raw data files were converted into mzXML format and then processed with the XCMS, CAMERA, and metaX toolbox implemented in the R software.