Overuse of antibiotics has become the major factor for the emergence and dissemination of multi-drug resistant strains of several groups of microorganisms and this lead to search for agents that may have antibacterial effects. Vitamin E emerged as an essential, fat-soluble nutrient in the human body and it is essential, because the body cannot manufacture its own vitamin E, so foods and supplements must provide it. The aim of the present study was to evaluate the effect of vitamin E against pathogenic bacteria. Gram positive and gram negative bacteria were selected as the test microorganisms based on their importance in infections. In this study vitamin E used in four concentrations (50,100,200,400) IU/ml. The agar diffusion method was used to determine antibacterial activity. Results showed that gram negative bacteria were shown to be more resistant than gram positive bacteria. The resistance of gram negative bacteria towards antibacterial substances may be related to lipopolysaccharides in their outer membrane.