Flax is one of the oldest oil crops, but only since the end of the twentieth century nutritional use of its whole seeds and flour has been resumed. This crop has been evaluated for its oil fatty acid composition, content of sterols and tocopherols, carbohydrate composition of mucilage, but a comprehensive study has never been carried out, so the aim of the work was to identify differences in the metabolomic profiles of flax lines contrasting in color and size of seeds. The biochemical composition of seeds from 16 lines of the sixth generation of inbreeding was tested using gas chromatography coupled with mass spectrometry. In total, more than 90 compounds related to sugars (78% of the identified substances), free fatty acids (13%), polyatomic alcohols (5%), heterocyclic compounds, free amino acids, phytosterols and organic acids (no more than 2.5% in total) were identified. Statistical analyses revealed six main factors. The first is a factor of sugar content; the second one affects most of organic acids, as well as some free fatty acids, not related to reserve ones, the third factor is related to compounds that play a certain role in the formation of "storage" substances and resistance to stress, the fourth factor is influencing free polar amino acids, some organic and free fatty acids, the fifth one is a factor of phenolic compounds, the sixth factor combined substances not included in the first five groups. Factor analysis made it possible to differentiate all 16 lines, 10 of which occupied a separate position by one or two factors. Interestingly, the first two factors with the highest loads (20 and 15% of the total variability, respectively) showed a separate position of the gc-432 line, which differed from the others, not only by chemical composition, but also by the phenotype of the seeds, while gc-159 differed from the rest ones by the complex of organic acids and other substances taking about 1% of the extracted substances of the seed. Thus, the analysis of metabolomic profiles is promising for a comprehensive assessment of the VIR flax genetic collection, which has wide biochemical diversity.