LED *Shared senior authors. light sources have a discontinuous light spectrum with a prominent ‘blue’ peak between 450 and 470 nm that influences non-image forming responses in humans. We tested an LED lighting solution mimicking a daylight spectrum on visual comfort, circadian physiology, daytime alertness, mood, cognitive performance and sleep. Fifteen young males twice spent 49 hours in the laboratory under a conventional-LED and under a daylight-LED condition in a balanced cross over design flanked by a baseline and a post-light exposure night. Despite different light spectra, the photopic lux and the correlated colour temperature of the lighting were the same for both LEDs. The colour rendering index and the melanopic strength were 25.3% and 21%, respectively, higher for the daylight LED than the conventional LED. The volunteers had better visual comfort, felt more alert and happier in the morning and evening under daylight LED than conventional LED, while the diurnal melatonin profile, psychomotor vigilance and working memory performance were not significantly different. Delta EEG activity (0.75–4.5 Hz) was significantly higher after daylight-LED than conventional-LED exposure during the post-light exposure night. We have evidence that a daylight-LED solution has beneficial effects on visual comfort, daytime alertness, mood and sleep intensity in healthy volunteers.