The aim of our study was the identification of two categories of bioactive molecules, characteristic to functional oils: sunflower, soybean, rapeseed, linseed, and hempseed oil, processed from seeds in Romania. Two separation and purification techniques, column chromatography and thin layer chromatography, were applied and the profiles of fatty acids from total lipids and phytosterol esters fraction was determined by GC-FID. Transesterification procedures were used for derivatization of fatty acids for GC analysis. The main fatty acids from the studied oil samples were: oleic, linoleic and palmitic fatty acids. Hemp oil is one of the few seed oils that contains both of omega-6 and omega-3, gamma-linolenic acid being one of relatively rare fatty acids. A 3.07% content of -linolenic acid was found. Hemp oil proved to be the most rich in phytosterols, been recommended as a functional food.