Introduction: The training and competition alter the metabolism and body composition greatly, especially those of long duration in one or several stages, where the energy becomes to be deficient or the metabolic profile changes (German et al., 2003; Weitkunat et al., 2012; da Silva et al.., 2013). There is a little research, and this is the aim of this study, focusing on analyzing the metabolomic response and profile of a cycling test (1000km non-stop) with an individualized nutritional intervention to assess as a response of the organism. Methods: A well-trained long-distance cyclist developed 1000km non-stop of cycling in a road circuit (4.7km) with a 40-hours limit. The nutritional and fluid intake was registered, during the entire test (pre, during -6 measures- and at the end) bioimpedance analysis -BIA- (Z-Metrix®, Bioparhom) were used to determine changes in body mass, skeletal muscle mass and fat mass. In order to quantify hydration status we measured intracellular water volume and urinary specific gravity and urine samples were collected for 1H NMR (PCA model). Results: The body composition changed from the beginning until the end (decreased: fat mass, skeletal muscle mass and intracellular water volume) and the urine specific gravity changed too (1005 g/mL to maximum of 1030 g/mL and 1020g/mL at the end). The PCA showed differences in global metabolic profiles in the whole event, from pre-, during and post- in the different PCA spaces. Conclusion: Our preliminary results suggest that is important to analyze the principal metabolites that contribute to change the metabolomics profile (not only in urine samples) during a long-distance sport event to avoid energy and hydration deficiency effects as well as to assess the catabolic effects that occur in the athlete.