Abstract: Objective: To investigate the changes of energy metabolism regulation in high-level female football players before and after a match through the liquid chromatography-mass spectrometry(LC-MS), so as to provide theoretical reference for the assessment of female football players' performance and monitoring the training. Methods: 10 female athletes who participated in the semi-finals of the 2021 College Women's Super Champion League were participated in this study. The urine samples in the morning, 30 minutes before and 30 minutes after the competition were collected. Data were converted, peak detected, and normalized for multivariate statistical analysis. Results: Significant differences in urine metabolite chromatograms were observed at before and after intermittent high-intensity interval training. Compared with before exercise, metabolites including lactate, creatine,pyruvate, creatinine, L-valine, tryptophan and alanine were significantly up-regulated after exercise(VIP>1.3 and P<0.01), the adenine, β-glucose, tyrosine, citric acid and succinic acid were decreased significantly(VIP>1.4 and P<0.01). Conclusions: 1) The metabolic changes observed in high-level female soccer players after intense matches mainly involve the ATP-CP system,glycolysis system, aerobic oxidation system, neural regulation system, and amino acid metabolism system; 2) Metabolomics methods can provide more comprehensive and targeted data analysis and guidance for athletes' diet and nutrition supplementation during preparation and pre-competition period as well as evaluation of their performance.