The purpose of this study was to develop the separation process removing sediment in sea buckthorn seed ethanol extract, and to investigate the antioxidant and antibacterial behavior for the sediment removed ethanol extract (SBS extract). Sediment such as low solubility materials and oil component was simply removed by filtering after storing the crude extract containing celite for 16 h in a refrigerator at 4 ℃. The SBS extract displayed strong antioxidant activity in the stability test as a function of both storage time and temperature when compared to vitamin C and BHA (Butylated hydroxyanisole) as typical antioxidant ingredients. The bactericidal ratio of the SBS extract against E. coli increased with an increase in the addition of the test agent, and this was particularly significant at concentrations greater than 1,000 μg/ml addition. The bactericidal ratio of 1,000 μg/ml SBS extract against B. subtilis, S. aureus, P. aerusinosa and E. coli was 96.4, 98.6, 91.5, and 52.2% after 2 h, and 98.2, 99.7, 99.3, and 82.6% after 4 h, respectively. These results suggest that the SBS extract may be used as a functional food, pharmaceutical, and cosmetic agent.