The aim of the present study was to evaluate time-on-task effects on subjective fatigue in two different tasks of varying monotony during night-time testing (20:00 to 4:00 hours) in a sleep deprivation intervention. The experiment included eight test runs separated by breaks of approximately 20 min. Twenty healthy volunteers performed a driving simulator and the Mackworth clock vigilance task in four of the test runs each. Sequence of tasks was varied across subjects. Before and after each task, subjective sleepiness was assessed by means of the Karolinska sleepiness scale and subjective fatigue was rated on the Samn-Perelli checklist. Fatigue and sleepiness significantly increased over the course of the night. Both tasks led to an increase in fatigue and sleepiness across test runs. However, this time-on-task effect was larger in the vigilance than in the driving simulator task. It is important to note that fatigue and sleepiness in one test run were not influenced by the task performed in the preceding test run, that is there were no cross-over effects. The results suggest that time-on-task effects superimpose circadian and sleep-related factors affecting fatigue. They depend on the monotony of the task and can be quantified by means of a design including separate test runs divided by breaks.