There is little information on the impact of hydration status on the psychological response to exercise despite potential implications for adherence to an exercise programme and for overall health and fitness. We investigated initial hydration status, fluid balance, and psychological responses associated with a typical recreational exercise session in healthy adults. Fifty-two participants performed a freely chosen gymnasium-based exercise session at a fitness centre, with ad libitum access to fluids. Urine samples were collected on arrival for analysis of osmolality. Sweat loss was estimated from the change in body mass after correction for fluid intake and urinary losses. Subjective psychological ratings were recorded before and after exercise. Pre-exercise urine osmolality was above 900 mOsmol · kg(-1) (used as a threshold for hypohydration) in 37% of participants. Fluid intake during exercise was 390 ± 298 mL, while estimated sweat loss was 794 ± 391 mL. The percentage change from pre-exercise body mass was -0.62 ± 0.20%. Physically active adults who arrived to take part in exercise hypohydrated reported more negative changes in psychological affect in response to their subsequent freely chosen recreational exercise session than those classified as euhydrated prior to exercise (-0.2 ± 0.7 vs. 0.8 ± 0.7; P < 0.005).