Background: A greater understanding of exercise physiology and biochemistry is required for the sport horse disciplines, including show jumping. Conditioning of horses for show jumping is empirical because they are primarily trained on flat ground, however the equivalent workload between jumping and flat work is currently unknown. The objectives of the study were therefore to compare the physiological demands of Warmblood show jumpers over a standardized 1.10 m course vs a 600 m standardized incremental exercise test on flat ground, and to report reference field test values for competitive show jumping horses. In this prospective field study, 21 healthy, actively competing Warmblood show jumping horses were assessed to determine physiological variables after a standardized jumping course at 6.4 m/s (average speed) and track standardized incremental exercise test at 5 m/s, 8 m/s and 11 m/s. Heart rate, velocity, blood lactate, blood pH, pCO2, bicarbonate, PCV and TP concentrations were recorded. V200, V170 and VLa4 were calculated. Parametric statistics were performed on analysis of all 21 horses' variables. Results: Contrary to exercise at 5 m/s and 11 m/s, cantering at 8 m/s did not induce any significant difference in blood lactate, mean heart rate or mean venous blood pH compared to after completion of the jumping course. Conclusions: Jumping a 1.10 m course demands a statistically similar workload to cantering around a flat track at 8 m/s. This study will help to test fitness and design conditioning programs for Warmblood show jumping horses.