Untargeted metabolomics analysis
HEAT
The collected plasma samples before and after exercise were subjected to untargeted metabolomics analysis using High-performance Liquid Chromatography. In addition, each sample was analyzed using Q Exactive Focus Orbitrap Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC-MS/MS) (Thermo Scientific, Waltham, USA) operated in the full-scan mode. The electrospray ionization source parameters were conducted as follows: nebulising gas flow of 3 L/min; the heating gas flow of 10 L/min; interface temperature of 500 °C; DL temperature of 250°C; heat block temperature of 400°C; and drying gas flow of 10 L/min. The obtained raw data was analyzed using Compound Discoverer 3.3 software.In the Compound Discover 3.3 software, high-quality (S/N > 10) metabolic signals from the MS2 spectral information were first compared with the database, and the metabolic signals were annotated in batches. Metabolic signals that were unmatched by the information in the database were identified by querying the MS2 spectral data from past literature or searching in other online databases, such as METLIN and MassBank. Compounds with standards were identified by comparing the Retention Time (RT), accurate m/z values, and fragmentation patterns with those obtained by injecting the standards under similar conditions.