Atopic dermatitis has been related to a disturbed metabolism of polyunsaturated fatty acids (PUFAs). We tested whether the PUFA composition of breast milk differs significantly between mothers with atopic dermatitis, mothers with other types of atopy, and nonatopic mothers. We also investigated whether differences in diet can explain possible observed differences. Mothers with current or previous asthma (n = 396) were divided into 3 groups according to history of atopic dermatitis and allergic rhinitis. Breast-milk samples were collected from 314 women approximately 3 wk after delivery. The habitual diet of the women was assessed with food-frequency questionnaires in the 25th week of gestation (n = 207). Breast-milk samples and simultaneous dietary data from 14 nonatopic mothers were used for comparison. Compared with the milk of nonatopic mothers, that of atopic mothers had significantly higher concentrations of 22:5n-6 and lower concentrations of 20:5n-3; moreover, 20:4n-6/20:5n-3, 22:5n-6/22:6n-3, and long-chain n-3 PUFA/18:3n-3 were shifted toward n-6 PUFA and 18:3n-3 in nonatopic and atopic mothers, respectively. No differences in breast-milk PUFA composition were evident between the subject groups. The diets of the groups differed only slightly with respect to protein intake. However, the PUFA composition of the breast milk was associated with diet and time of milk sampling, and the above difference in milk PUFAs disappeared when those factors were taken into account. Our data do not support the possibility that the fatty acid composition of breast milk is affected by atopic dermatitis or atopy in general, because most differences in breast-milk PUFA composition appear to be explained by the diet.