Studies on HPA axis regulation in burnout revealed heterogeneous results, possibly due to different psychometric and endocrine measurements, heterogeneous samples or small sample sizes. In the present study, the relationship between salivary cortisol during the day (four time points: 0700 h, 1130 h, 1730 h, and 2000 h) and burnout as well as vital exhaustion was investigated in a large sample of 279 nurses. Burnout was measured using the Maslach Burnout Inventory (MBI), which includes scales for emotional exhaustion, depersonalization, and personal accomplishment. A burnout criterion was assumed to be fulfilled, when any of the MBI scales was above the norm. Subjects with two burnout criterions fulfilled (N=18) were characterized by a higher cortisol release over the day compared to those reporting only one criterion (N=77) or no burnout (N=181) (ANOVA, p=.015). On the other hand, subjects who reported high levels of vital exhaustion did not differ from those who did not report signs of vital exhaustion. These findings provide further evidence for HPA axis dysregulation in burnout.