This study investigated the repeated-sprint ability (RSA) physiological responses to a standardized, high-intensity, intermittent running test (HIT), maximal oxygen uptake (VO(2) (max)) and oxygen uptake (VO(2)) kinetics in male soccer players (professional (N = 12) and amateur (N = 11)) of different playing standards. The relationships between each of these factors and RSA performance were determined. Mean RSA time (RSA(mean)) and RSA decrement were related to the physiological responses to HIT (blood lactate concentration ([La(-)]), r = 0.66 and 0.77; blood bicarbonate concentration ([HCO(3)-]), r = -0.71 and -0.75; and blood hydrogen ion concentration ([H(+)]), r = 0.61 and 0.73; all p < 0.05), VO(2) (max) (r = -0.45 and -0.65, p < 0.05), and time constant (tau) in VO(2) kinetics (r = 0.62 and 0.62, p < 0.05). VO(2) (max) was not different between playing standards (58.5 +/- 4.0 vs. 56.3 +/- 4.5 mL.kg(-1).min(-1); p = 0.227); however, the professional players demonstrated better RSA(mean) (7.17 +/- 0.09 vs. 7.41 +/- 0.19 s; p = 0.001), lower [La-] (5.7 +/- 1.5 vs. 8.2 +/- 2.2 mmol.L(-1); p = 0.004), lower [H+] (46.5 +/- 5.3 vs. 52.2 +/- 3.4 mmol.L(-1); p = 0.007), and higher [HCO3-] (20.1 +/- 2.1 vs. 17.7 +/- 1.7 mmol.L(-1); p = 0.006) after the HIT, and a shorter in VO2 kinetics (27.2 +/- 3.5 vs. 32.3 +/- 6.0 s; p = 0.019). These results show that RSA performance, the physiological response to the HIT, and differentiate between professional- and amateur-standard soccer players. Our results also show that RSA performance is related to VO(2) max, tau, and selected physiological responses to a standardized, high-intensity, intermittent exercise.