The circadian rhythm regulates arousal levels throughout the day and determines optimal periods for engaging in mental activities. Individuals differ in the time of day at which they reach their peak: Morning-type individuals are at their best in the morning and evening types perform better in the evening. Performance in recall and recognition of non-facial stimuli is generally superior at an individual’s circadian peak. In two studies (Ns = 103 and 324), we tested the effect of time-of-testing optimality on eyewitness identification performance. Morning- and evening-type participants viewed stimulus films depicting staged crimes and made identification decisions from target-present and target-absent lineups either at their optimal or non-optimal time-of-day. We expected that participants would make more accurate identification decisions and that the confidence-accuracy and decision time-accuracy relationships would be stronger at optimal compared to non-optimal time of day. In Experiment 1, identification accuracy was unexpectedly superior at non-optimal compared to optimal time of day in target-present lineups. In Experiment 2, identification accuracy did not differ between the optimal and non-optimal time of day. Contrary to our expectations, confidence-accuracy relationship was generally stronger at non-optimal compared to optimal time of day. In line with our predictions, non-optimal testing eliminated decision-time-accuracy relationship in Experiment 1.