Objective Mental fatigue, a poorly understood symptom of sports-related concussion, ideally requires assessment across multiple modalities. Our study aimed to examine mental fatigue effects among 10 neurologically normal, athletically active students undergoing typical concussion testing. It is our intention to ultimately address the question whether fatigue effects due to mild traumatic brain injury (mTBI) may become confounded with fatigue effects due to testing effort. Methods Fourteen athletically active and neurologically normal volunteers were initially recruited from Penn State University. Self-reported fatigue, neuropsychological performance, and electroencephalographic (EEG) activity were measured throughout the whole testing duration. EEG measures in frequency domain (e.g., relative power of theta, alpha, and beta bands) were examined over the course of neuropsychological (NP) test administration. Results Predicted fatigue effects over the course of testing included: (a) increased self-reported fatigue; (b) increased errors on the Stroop Interference Test; (c) significantly increased relative power of theta activity during the Stroop Interference Test in frontal–central and parietal regions; and (d) migration of alpha activation from the occipital to anterior (left parietal and pre-central) regions during the Stroop Interference task administered at the beginning compared with the end of testing. Conclusions Results supported predictions related to subjective fatigue and cognitive performance and offered partial support for predictions related to EEG activation patterns over the course of administering the NP testing. Significance Neurologically intact and athletically active college students demonstrate effects related to fatigue after undergoing a typical sports concussion assessment battery, including an increase in subjectively experienced fatigue, a decrease in cognitive task performance accuracy and associated modulations in EEG activity. This finding should be considered by clinical practitioners while evaluating the symptoms of concussion and making a decision regarding the return-to-sport participation.