The aim of this study was to investigate alertness and cognitive performance immediately after and until the end of the night shift after taking a 120-min monophasic nap (One-nap) or a split 90-min and 30-min nap (Two-nap) during a 16-h simulated night shift, and the relationship between sleep quality and both alertness and performance. This study was performed in 41 females. Among them, 15 participants were included in the No-nap group, 14 in the One-nap group (22:00–00:00), and 12 in the Two-nap group (22:30–00:00 and 02:30–03:00). Participants were tested every hour from 16:00 to 09:00 for performance on the Uchida-Kraepelin test, as well as for subjective feelings of fatigue and drowsiness, body temperature, and heart rate variability. The shorter the sleep latency, the worse the alertness immediately after the 90-min nap. The 120-min and 30-min naps also revealed that prolonged total sleep time led to increased fatigue and drowsiness upon awakening. From 04:00 to 09:00, in the No-nap and One-nap groups, fatigue was higher than in the Two-nap group. The One-nap and Two-nap groups did not show improved morning performance. These results suggest that a split nap might improve drowsiness and fatigue during a long night shift.