The cardiac regulation effects of a mental task added to regular office work are described. More insight into the time evolution during the different tasks is created by using time-frequency analysis (TFA). Continuous wavelet transformation was applied to create time series of instantaneous power and frequency in specified frequency bands (LF 0.04-0.15 Hz; HF 0.15-0.4 Hz), in addition to the traditional linear heart rate variability (HRV) parameters. In a laboratory environment, 43 subjects underwent a protocol with three active conditions: a clicking task with low mental load and a clicking task with high mental load (mental arithmetic) performed twice, each followed by a rest condition. The heart rate and measures related to vagal modulation could differentiate the active conditions from the rest condition, meaning that HRV is sensitive to any change in mental or physical state. Differences between physical and mental stress were observed and a higher load in the combined task was observed. Mental stress decreased HF power and caused a shift toward a higher instantaneous frequency in the HF band. TFA revealed habituation to the mental load within the task (after 3 min) and between the two tasks with mental load. In conclusion, the use of TFA in this type of analysis is important as it reveals extra information. The addition of a mental load to a physical task elicited further effect on HRV parameters related to autonomic cardiac modulation.