The aim of this study was to assess the effect of gamma irradiation treatment at doses of 0, 1, 2 and 3 kGy, storage time of fruits for 0, 30 and 45 days and storage time of oils for 0, 6 and 12 months on fatty acid profile of olive oil produced in Syria. Results of this study showed that fatty acid composition was different based on irradiation dose and storage time of fruits and oils. Small change were observed in saturated, monounsaturated and polyunsaturated fatty acid components due to irradiation and storage. However, the changes in fatty acids composition of oil extracted from irradiated and stored fruits or oils were sometimes significant (P<0.05). The composition of fatty acids of analyzed oil samples was determined as palmitic acid (C16:0) from 13.25% to 14.60%; palmitoleic acid (C16:1) from 0.82% to 1.43%; stearic acid (C18:0) from 2.43% to 3.83%; oleic acid (C18:1) 68.95% to 72.64%; linoleic acid (C18:2) 7.65% to 11.93%; Linolenic acid (C18:3) from 0.41% to 1.09%. The present study demonstrated that the effect of gamma irradiation, fruit and oil storage on the fatty acid profile of virgin olive oil (VOO) was minimized.