The objective of this study was to determine if children with phenylketonuria (PKU) have lower fatty acid concentrations in total erythrocyte lipid due to the phenylalanine restricted diet therapy compared to healthy control subjects. Dietary intake and fatty acid concentrations in total erythrocyte lipid were measured in twenty-one subjects (≤6 years of age) with PKU and twenty-three control children. Subjects with PKU had significantly lower protein and significantly higher polyunsaturated fat intake compared to controls. Subjects with PKU had significantly lower concentrations in total erythrocyte lipid of the sum of the ω-3,ω-6, saturated and polyunsaturated fatty acids. Concentrations of fatty acids among subjects with PKU were lower than control subjects but no subject with PKU exhibited any signs or symptoms suggestive of essential fatty acid deficiency, thereby suggesting that subjects with PKU in this cohort have normal and adequate essential fatty acid concentrations in total erythrocyte lipid.