This study aims to investigate the effects of workplace noise on neural activity and alpha asymmetries of the prefrontal cortex (PFC) during mental stress conditions. Workplace noise exposure is a pervasive environmental pollutant and is negatively linked to cognitive effects and selective attention. Generally, the stress theory is assumed to underlie the impact of noise on health. Evidence for the impacts of workplace noise on mental stress is lacking. Fifteen healthy volunteer subjects performed the Montreal imaging stress task in quiet and noisy workplaces while their brain activity was recorded using electroencephalography. The salivary alpha-amylase (sAA) was measured before and immediately after each tested workplace to evaluate the stress level. The results showed a decrease in alpha rhythms, or an increase in cortical activity, of the PFC for all participants at the noisy workplace. Further analysis of alpha asymmetry revealed a greater significant relative right frontal activation of the noisy workplace group at electrode pairs F4-F3 but not F8-F7. Furthermore, a significant increase in sAA activity was observed in all participants at the noisy workplace, demonstrating the presence of stress. The findings provide critical information on the effects of workplace noise-related stress that might be neglected during mental stress evaluations. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.