Work and intellectually fatiguing environments can significantly influence the health of individuals, which is strictly bound to motor efficiency. In particular, desk workers and university students may have a sedentary lifestyle and a condition of mental fatigue caused by daily routine, which could impair motor efficiency. The assessment is a starting point for enhancing awareness of the individual’s psychophysical condition through the perception of one’s body motor efficiency, motivating to move towards improvement. This way, a submaximal test based on perceived exertion was developed (Cubo Fitness Test, CFT) and validated in previous studies. Hence, two further studies were employed to enhance the consistency and accuracy of this instrument in different conditions. The first study investigated the internal responsiveness of CFT, evaluating if mental fatigue could affect motor efficiency. The second study investigated which perceived intensity (weak, moderate, strong, or absolute maximum) could be more reliable for applying the CFT (as previous research focused the investigation only on moderate intensity). In the first investigation, participants assessed two stimuli (mental fatigue induced with a Stroop color-word task and a neutral condition based on the vision of a documentary) lasting 60 min each. The quality of psychophysical recovery (total quality recovery) and the mood state (Italian Mood State questionnaire) were evaluated before the stimuli. After the fatiguing or the neutral task, the mood state was newly assessed, together with the evaluation of the workload’s characteristics (Nasa TLX) and the CFT motor efficiency. In the second investigation, participants had to perform CFT twice for each at different intensities of Borg’s Scale of perceived exertion. Researchers successfully requested to fill out the NASA TLX questionnaire regarding the perceived workload characteristics of CFT, and the reliability of each intensity was assessed. Results seem to enhance the consistency and the accuracy of the instrument. Indeed, findings evidenced that CFT is not influenced by mental fatigue conditions typical of the intellectual work of desk workers and university students for which this test was specifically conceived. Moreover, moderate and strong perceived intensity are the most adequate conditions to assess motor efficiency in these populations.