A standardized untargeted serum metabolomics workflow using LC-QTOF-MS/MS revealed that acute sparring in elite boxers was associated with glycolysis and gluconeogenesis metabolite changes, while 24-h recovery was associated with sulfur metabolism, with phosphatidylinositol PI(16:0/18:2(9Z,12Z)) and thiosulfate serving as discriminating metabolites for the respective timepoints.
Key Findings
Results
Acute sparring in elite boxers was associated with changes in metabolites related to glycolysis and gluconeogenesis.
Study involved seven elite male boxers.
Samples were collected at three timepoints: before sparring, immediately after sparring, and 24 h after sparring.
Metabolite profiling was performed using liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (LC-QTOF-MS/MS).
Glycolysis and gluconeogenesis pathway changes were identified in the immediate post-sparring state.
Results
The 24-h post-sparring recovery timepoint was associated with sulfur metabolism changes.
Sulfur metabolism pathway changes were specifically identified at the 24-h post-sparring timepoint.
Thiosulfate was identified as a metabolite associated with the 24-h recovery state.
This temporal distinction suggests different metabolic processes dominate acute versus short-term recovery phases.
The 24-h timepoint was one of three standardized collection points in the study design.
Results
Phosphatidylinositol PI(16:0/18:2(9Z,12Z)) showed strong discrimination of the immediate post-sparring state.
This specific lipid metabolite was identified through multivariate statistical analysis as a key discriminating feature.
The finding emerged from an untargeted metabolomics approach, not a hypothesis-driven targeted assay.
Discrimination was assessed in a cohort of seven elite male boxers.
Multivariate statistical analysis and pathway interpretation were included as components of the standardized workflow.
Methods
A standardized untargeted serum metabolomics workflow was developed and validated for reproducible profiling of exercise-related metabolic changes in small cohorts.
The workflow included standardized sample collection, pooled quality-control (QC) monitoring, metabolite profiling, multivariate statistical analysis, and pathway interpretation.
Pooled QC monitoring was emphasized as a key component for ensuring reproducibility.
The protocol is explicitly intended for controlled small-cohort studies of acute exercise and short-term recovery.
The paper is published as a JoVE (Journal of Visualized Experiments) protocol article, indicating a methods-focused contribution.
The workflow was applied to samples from seven elite male boxers as proof-of-concept.
Background
Conventional physiological measurements do not fully capture the rapid metabolic changes induced by elite boxing.
This limitation of conventional measurements was stated as a primary motivation for developing the metabolomics workflow.
LC-QTOF-MS/MS-based untargeted metabolomics was proposed as a more comprehensive alternative.
The approach enables simultaneous profiling of a broad range of serum metabolites without prior hypothesis about specific analytes.
What This Means
This research describes a detailed laboratory method for measuring hundreds of chemical compounds in blood samples from elite boxers to understand how their bodies respond to a sparring session. Using a highly sensitive instrument called a liquid chromatography-mass spectrometer, the researchers analyzed blood taken from seven elite male boxers before a sparring session, right after it ended, and again 24 hours later. By comparing the chemical profiles at these three timepoints, they could identify which metabolic processes were most active at each stage of exercise and recovery.
The study found that immediately after sparring, the boxers' blood showed signs of increased activity in energy-producing pathways — specifically those involved in breaking down and producing glucose (glycolysis and gluconeogenesis). A specific fat-related molecule called phosphatidylinositol PI(16:0/18:2) was particularly useful for distinguishing the post-sparring state from the pre-sparring baseline. At the 24-hour recovery mark, the chemical signature shifted toward sulfur metabolism, with a compound called thiosulfate being notably elevated, suggesting a different set of biological repair and recovery processes were underway by that point.
This research suggests that untargeted metabolomics — measuring many metabolites at once without deciding in advance what to look for — can reveal aspects of athletic stress and recovery that standard fitness tests might miss. The authors emphasize that their carefully standardized protocol, including quality-control checks throughout sample processing, makes the results reproducible and trustworthy even in a small group of athletes. The method is intended as a tool for sports scientists and physiologists studying how the body responds to intense exercise in controlled research settings.
Check Your Own Numbers
Upload your bloodwork. We'll cross-reference your results against this study and 4,700 others.
Sun P, Zhang D, Chen Q, He M, Li B. (2026). A Standardized Metabolomics Protocol For Analyzing Exercise-Induced Metabolic Shifts In Elite Boxers By Liquid Chromatography-Mass Spectrometry.. Journal of visualized experiments : JoVE. https://doi.org/10.3791/70719