The color and light have effect on physiology, psychology, cognitive performance and hormone production in the human. In this paper, we aim to study red and blue color with their effect on brain activity, cardiovascular activity, emotion and saliva hormone. Each participant was testing in red and blue room. Participant was measure the EEG, ECG, SpO2, pulse rate, saliva hormone (melatonin, cortisol, testosterone, progesterone, estradiol and DHEAS), and emotion. The results showed that red room regulates the tension, anger, vigor and confusion moods which stimulate significant increasing of the pulse rate and SpO2 and cortisol levels but blue room regulates the depression and fatigue. The brain activity of high beta wave (25–30 Hz) was high at the occipital lobe and little high at frontal lobe in red room but the blue room was high at frontal lobes only. The coherence of high beta wave was not difference in both rooms. The Gonadal and melatonin hormone were not clearly changing in red and blue room.