Hydrolysis of vegetable oils (Olive, corn, peanut, sesame, flaxseed, soy, canola, garlic, sunflower, almond, castor bean oils) and beef marrow bone oil by lipase was studied. The enzyme was capable of generating free fatty acids from all oils tested. The higher hydrolytic activity of the enzyme was towards olive (18.0 IU) and soybean (17.8 IU) oils. The average percentage of essential fatty acids generated from hydrolysis of the oils was 32.92% of omega 9 (as oleic acid C18:1), 26.24% of omega 6 (linoleic C18:2), and 5.86% of omega 3 (such as α-linolenic acid C18:3). Comparison between chromatographic profile of the oils and its enzymatic hydrolysate showed a good equivalence, stressing the applicability of these vegetable substrates under the action of lipase from produce essential fatty acids, being more efficient production of α-linolenic acid from flaxseed oil, linoleic acid from sunflower oil, and oleic acid from olive.