Flax, Linum usitatissimum, cultivars are grown throughout the world. Flax oil is a dietary source of polyunsaturated fatty acids, vitamin E, as well as phospholipids, sterols, and phenolic acids. Linseed plays a pivotal role in protecting cells from oxidative damage associated diseases, i.e., atherosclerosis, neurodegenerative disorders, cancer, and inflammation. In this study, two groups of L. usitatissimum seeds were used to evaluate and compare the content and composition of tocochromanols (vitamin E) and fatty acids. Group I included accessions originating from Poland and the Ukraine, while Group II encompassed worldwide flax cultivars (such as from the United States, Argentina, and Italy). A comparison of the tocochromanol profiles showed a higher content in Group I, although there were no significant differences in tocopherol content and composition between the genotypes within this group. All accessions in Groups I and II contained γ-tocotrienol and plastochromanol-8, which confirms the high nutritional value of flaxseeds. The composition of fatty acids varied depending on the varieties, with linolenic acid showing the greatest discrepancy. Based on the tocochromanol content and fatty acid composition, we conducted a principal component analysis (PCA) and cluster analysis, which revealed a greater similarity among the accessions in Group I. An analysis of the tocochromanol and fatty acid composition of flaxseeds is important from an agronomic and medicinal perspective and can be used to select the most appropriate flax cultivar.