A Standardized Metabolomics Protocol For Analyzing Exercise-Induced Metabolic Shifts In Elite Boxers By Liquid...

Elite boxing induces rapid metabolic changes that are not fully captured by conventional physiological measurements. A standardized untargeted serum metabolomics workflow based on liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (LC-QTOF-MS/MS) was applied to samples collected before sparring, immediately after sparring, and 24 h after sparring in seven elite male boxers. The workflow included standardized sample collection, pooled quality-control monitoring, metabolite profiling, multivariate statistical analysis, and pathway interpretation. Acute sparring was associated with changes in metabolites related to glycolysis and gluconeogenesis, whereas the 24-h timepoint was associated with sulfur metabolism. Phosphatidylinositol PI(16:0/18:2(9Z,12Z)) showed strong discrimination of the immediate post-sparring state, and thiosulfate was associated with the 24-h recovery state. These findings support the use of this workflow for reproducible profiling of exercise-related metabolic changes in this cohort of seven elite male boxers. The protocol is intended for controlled small-cohort studies of acute exercise and short-term recovery and emphasizes reproducible sample handling, pooled quality-control monitoring, and interpretable downstream analysis.