Objective We assessed operator performance during a real-time reactive telerobotic lunar mission simulation to understand how daytime versus nighttime operations might affect sleepiness, performance, and workload. Background Control center operations present factors that can influence sleepiness, neurobehavioral performance, and workload. Each spaceflight mission poses unique challenges that make it difficult to predict how long operators can safely and accurately conduct operations. We aimed to evaluate the performance impact of time-on-task and time-of-day using a simulated telerobotic lunar rover to better inform staffing and scheduling needs for the upcoming Volatiles Investigating Polar Exploration Rover (VIPER) mission. Methods We studied seven trained operators in a simulated mission control environment. Operators completed two five-hour simulations in a randomized order, beginning at noon and midnight. Performance was evaluated every 25 minutes using the Karolinska Sleepiness Scale, Psychomotor Vigilance Task, and NASA Task Load Index. Results Participants rated themselves as sleepier (5.06 ± 2.28) on the midnight compared to the noon simulation (3.12 ± 1.44; p < .001). Reaction time worsened over time during the midnight simulation but did not vary between simulations. Workload was rated higher during the noon (37.93 ± 20.09) compared to the midnight simulation (32.09 ± 21.74; p = .007). Conclusion Our findings suggest that work shifts during future operations should be limited in duration to minimize sleepiness. Our findings also suggest that working during the day, when distractions are present, increases perceived workload. Further research is needed to understand how working consecutive shifts and taking breaks within a shift influence performance.