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We have been using “-Omics” sciences with classic laboratory analyses to understand the systemic metabolic and signaling changes induced by sport and exercise. We called this approach Sportomics. Our samples were collected in situ either during competitions or training to mimic the genuine challenges and conditions faced during sports. Non-targeted analysis (NTA) has opened the door to a new era of high-throughput exercise-induced metabolic research. In the present study, 30 male semi-professional soccer players were observed for two subsequent days. Both blood and urine samples were collected immediately pre-match and after matches. The most up-regulated prominent molecules were fatty acyls, carboxylic acids and derivatives, steroids and steroid derivatives. The most down-regulated molecules were fatty acyls, carboxylic acids and derivatives, as well as benzene and substituted derivatives. After metabolite identification and determining which metabolites were up- or down-regulated, we took the metabolites and grouped them into classes to examine the metabolic pathways involved with purine metabolism and to investigate hyperammonemia. To follow-up on our findings in urine, we used point-of-care instrument analysis to measure capillary blood metabolites. Glucose significantly increased by 35%, whereas urate increased by 16% and uremia by 17% without any changes in creatinemia. In the present study, we showed that hypoxanthine and related metabolites were up-regulated in urine after a soccer match, which suggested that AMP deamination was increased. In this study, we demonstrated several results through urine non-target mass spectrometry (NTMS) to understand exercise-induced changes during a soccer match using a Sportomics approach. These data together demonstrated that during a soccer game, there was an increase in ATP use provided by ADP synthesis via myokinase. Our data may show that the use of urinary metabolomics can be a less invasive way to follow the metabolism of athletes during exercise. We demonstrated that the use of NTMS may be useful for future studies that aim to design holistic interventions for improving athletic performance.