Graphical Abstract: The present paper describes biochemical (fatty oil) composition of Silybummarianum (L.) Gaertn. of Moldovan origin. The oil content of the seeds was approximately 25%. Linoleic acid (C18:2), an essential polyunsaturated fatty acid, is the most abundant (48.88%), followed by monounsaturated oleic acid (C18:1, 31.94%) and saturated palmitic acid (C16:0, 7.61%). The RP-HPLC analysis of tocopherols composition, showed as main components α-tocopherol (23.45 mg/100g) and γ–tocopherol (5.60 mg/100g). Based on the obtained results, it was shown that the extracted oil from milk thistle seeds is rich in essential fatty acids (about 50%) and tocopherols (29.09 mg/100g) and it can be used in food preparation.