Numerous research studies have formulated abstract assertions concerning how the human perception of color can stimulate a physiological response that enhances cognitive concentration. Within the framework of the color-in-context theory, interpretations and impacts of color are contingent on specific contexts; a single color can convey divergent or contrasting meanings in alternative settings. This pilot investigation seeks to enhance the experimental design for assessing the impact of different background colors using the U.S. Army Aeromedical Research Laboratory's Multi-Attribute Task Battery (USAARL MATB) simulation software. In this study, 8 participants undertook tasks of varying difficulty levels, with alterations to the background colors—specifically red, blue, grey, yellow, and green. To evaluate mental workload, pupillometry data, including average pupil diameter and average duration of fixation, were collected using the Tobii eye-tracking device. Additionally, subjective measures utilized in this study are the NASA task load index (TLX) and instantaneous self-assessment (ISA). The performance results derived from this research were anchored in accuracy and time metrics, and these were subjected to statistical tests aimed at ascertaining significant differences. The findings revealed noteworthy disparities in task difficulty levels concerning both performance outcomes and subjective assessments. Moreover, the physiological data exhibited significant differences as determined through paired tests. Notably, the three mental workload evaluation discrepancies can be attributed to the relatively small sample size, as anticipated.