Light, one of the key environmental components for both life and work, played significant role in subjective feelings (e.g. affect and comfort), but the exact effects and mechanisms were still to be determined. The present study screened thirty healthy adults (13 females, 22.45 +/- 3.26 years) and examined subjective affect and comfort under short-term white lights with different combination of correlated color temperature (CCT) and illuminance at different times of day (e.g. morning, afternoon, and evening). Our results showed a significant interaction between illuminance level and time-of-day on subjective comfort. Participants felt more comfortable under 50 lx and 100 lx instead of 500 lx in the evening, and more comfortable under 500 lx in the morning and afternoon. In addition, a positive correlation between illuminance and comfort in the morning and a negative correlation between them in the evening were found. No significant effect of CCT on any subjective feeling was revealed. Our results necessitate the consideration of time-of-day in understanding lighting effects and application of healthy lighting in daily life.