The aim of this study was to describe and compare the blood metabolic responses obtained after a single maximal exercise in elite and less-successful athletes and to investigate whether these responses are related to sprint performance. Eleven elite (ELI) and 14 regional (REG) long sprint runners performed a 300-m running test as fast as possible. Blood samples were taken at rest and at 4 minutes after exercise for measurements of blood lactate concentration [La] and acid-base status. The blood metabolic responses of ELI subjects compared to those of REG subjects for pH (7.07 ± 0.05 vs. 7.14 ± 1.5), sodium bicarbonate concentration ([HCO(3)(-)], 8.1 ± 1.5 vs. 9.8 ± 1.8 mmol·L(-1)), hemoglobin O(2) saturation (SaO(2)) (94.7 ± 1.8 vs. 96.2 ± 1.6%) were significantly lower (p < 0.05), and [La] was significantly higher in ELI (21.1 ± 2.9 vs. 19.1 ± 1.2 mmol·L(-1), p < 0.05). The 300-m performance (in % world record) was negatively correlated with pH (r = -0.55, p < 0.01), SaO2 (r = -0.64, p < 0.001), [HCO(3)(-)] (r = -0.40, p < 0.05), and positively correlated with [La] (r = 0.44, p < 0.05). In conclusion, for the same quantity of work, the best athletes are able to strongly alter their blood acid-base balance compared to underperforming runners, with larger acidosis and lactate accumulation. To obtain the pH limits with acute maximal exercise, coaches must have their athletes perform a distance run with duration of exercise superior to 35 seconds. The blood lactate accumulation values (mmol·L(-1)·s(-1)) recorded in this study indicate that the maximal glycolysis rate obtained in the literature from short sprint distances is maintained, but not increased, until 35 seconds of exercise.