Prolonged periods of demanding cognitive tasks lead to an exhausted feeling known as mental fatigue. The neural underpinnings of mental fatigue are still under exploration. In the present study, we aimed to identify neurophysiological indicators of mental fatigue by studying the time–frequency distribution of the event-related potentials (ERPs) measured in N=26 adults in nonfatigued versus fatigued states. We were interested in the frontal theta and occipital alpha variations, which have shown consistent relationships with mental fatigue in previous studies. Furthermore, we expected differential changes in left and right electrodes, in line with previously detected lateralization effects in cognitive tasks. Mental fatigue was induced by a sustained two-back verbal visual memory task for 125 min and assessed using the Chalder Fatigue Scale. We applied a high-resolution time–frequency analysis method called smoothed pseudo Wigner Ville distribution and used regional integrals as indicators for changing trends of signal energy. Results showed an increase in ERP frontal theta energy (P=0.03) and a decrease in occipital alpha energy (P=0.028) when participants became mentally fatigued. The change in frontal theta was more pronounced in left electrode sites (P=0.032), hinting toward a differential fatigue effect in the two hemispheres. The results were discussed on the basis of previous lateralization studies with memory tasks and interpreted as an indicator of a causal relationship between the sustained task execution and the physiological changes. Our findings also suggest that the ERP signal energy variations in frontal theta and occipital alpha might be used as neural biomarkers to assess mental fatigue.