Light exerts non-visual effects on a wide range of biological functions and behavior apart from the visual effect. Light can regulate human circadian rhythms, like the secretion of melatonin and cortisol. Light also has influence on body’s physiological parameters, such as blood pressure, heart rate and body temperature. However, human cognitive performance, alertness and mood under different lighting conditions have not been considered thoroughly especially for the complicated visual task like surgical operating procedure. In this paper, an experiment was conducted to investigate the cognition, alertness and mood of healthy participants in a simulated operating room (OR) in the hospital. A LED surgical lamp was used as the light source, which is mixed by three color LEDs (amber, green and blue). The surgical lamp is flexible on both spectrum and intensity. Exposed to different light settings, which are varied from color temperature and luminance, participants were asked to take psychomotor vigilance task (PVT) for alertness measurement, alphabet test for cognitive performance measurement, positive and negative affect schedule (PANAS) for mood measurement. The result showed the participants’ cognitive performance, alertness and mood are related to the color temperature and luminance of the LED light. This research will have a guidance for the surgical lighting environment, which can not only enhance doctors’ efficiency during the operations, but also create a positive and peaceful surgical lighting environment.