| Description |
Metabolomic analysis was performed using gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) following established protocols.Quality control samples were prepared by pooling equal volumes of each AH sample to assess the stability and reproducibility of the untargeted metabolic analysis. The current untargeted metabolic analysis was performed via GC-MS and LC-MS. For GC analysis, AH samples were analyzed on an Agilent 7890B gas chromatograph coupled to an Agilent 5977B MSD system (Agilent Technologies Inc., Santa Clara, CA, USA), and an HP-5MS fused-silica capillary column was utilized to separate the derivatives. For LC-MS, an ACQUITY UPLC I-Class system (Waters Corporation, Milford, MA, USA) coupled with a VION IMS QTOF Mass spectrometer (Waters Corporation, Milford, USA) was used to analyze the metabolic profiling in both ESI positive and ESI negative ion modes. An AC_x005f QUITY UPLC BEH C18 column was employed in both positive and negative modes.All raw data were imported into MS-DIAL software and Progenesis QI V2.3 software (Nonlinear, Dynamics, Newcastle, UK) for further identification processing. The missing values in the raw data were filled by using half of the minimum value. The identification of metabolites from GC-MS was based on the LUG database, while the detection of com pounds by LC-MS was performed based on the Human Metabolome Database (HMDB), Metlin, PMDB, and self-built databases. |
