Although the importance of the hypothalamic-pituitary-adrenal axis in emotional stress response is a well-established fact, the causal relationship between glucocorticoids and emotions remains unclear. Therefore, it was tried to elucidate this relationship by an experimental approach in 100 healthy volunteers. The effects of different oral dosages of cortisol (hydrocortisone; 20 and 40 mg) or placebo on emotional states were tested under stress and nonstress conditions in an independent group design. Furthermore, the time of drug application was varied in relation to the onset of the stress/nonstress conditions (1 vs. 2 h prior to exposure). Stress and nonstress conditions were induced by a film depicting violence and neutral scenes, respectively. The results indicate that the application of cortisol ameliorates emotional states irrespective of the time of drug administration and stress exposure. Also the dimensions of activity were increased by cortisol in all experimental conditions, except when applied 1 h before stress onset, indicating an adaptive negative feedback regulation preventing stress-induced overactivation.