Relationship between free oestrone and boar taint compounds in adipose tissue were studied in two groups of entire male pigs of different breeds. Group A consisted of 33 entire crossbred male pigs (dam Yorkshire and sire backcross Yorkshire × Wild Boar, generation seven). Group B consisted of 194 entire male pigs of crossbreeds between Swedish Hampshire (H) and Finnish Landrace (L), LH × H, H × LH, LH × LH (dam × sire). The measurements of free oestrone in adipose tissue were performed with a new method developed and validated in our laboratories. The standard curve was linear for concentrations of free oestrone ranging from 0.13 to 5.10 ng/g. The method exhibited parallelism of results between serial dilutions and a mean recovery of 97 ± 13.7%. Intra-assay variations for samples with concentrations of free oestrone from 0.67 to 2.08 ng/g were from 9.23 to 11.94%. Inter-assay variations for the samples with concentrations of free oestrone from 0.89 to 2.96 ng/g were from 3.78 to 10.11%. The levels of free oestrone in fat from group A were well correlated with fat levels of androstenone (r = 0.66; p < 0.001) and levels of oestrone sulphate in peripheral plasma collected at the same time as the fat (r = 0.74, p < 0.001). The levels of free oestrone in fat from group B were significantly correlated to fat levels of androstenone (r = 0.68, p < 0.001) and skatole (r = 0.29, p < 0.001). In group B, age-related differences in fat levels of free oestrone, androstenone and skatole were studied. Free oestrone and skatole levels increased simultaneously at the age of 22 week (p < 0.05 for both), and androstenone levels increased at the age of 26 week (p < 0.05). It was suggested that the levels of free oestrone in adipose tissue might be used for the evaluation of hormonal status of male pigs as an alternative to plasma levels of testicular hormones. The levels of free oestrone might be involved in the regulation of skatole levels in fat as indicated by both the simultaneous increases in skatole and free oestrone levels in fat and positive correlation between skatole and free oestrone.