>> When bioethanol and water are mixed at a proper ratio, phase separation can occur because of the immiscibility of biobutanol with water. Phase separation in bioethanol blends fuels is a major problem for gasoline vehicle users due to effect of octane number and component corrosion. Thus, in this study, the phase separation of bioethanol was examined effect of bioethanol blends (E3 (3 vo.% bioethanol in gasoline), E5 and E10) in presence of water. The effect were evaluated behavior with phase separation test, simulation test of fuel tank in gas station according to water addition volume and it was investigated change of water content, bioethanol content and octane number for gasoline phase in bioethanol blends (E3, E5 and E10) every 1 week after water addition. The E3 occurred phase separation more easily than the E5 and E10 in small water contents because solubility of water on ethanol content difference in gasoline-ethanol. It was kept a initial level of water content, bioethanol content, and octane number by repeated sample replacing in simulation test of fuel tank.