Milk from cows fed a low-fat diet high in cereals designed to stimulate fat synthesis de novo was lower in unsaturated fatty acids (21·3%) than milk from cows fed a diet high in fat, mainly from roasted soy beans (41·3% unsaturated fatty acids). Buttermilk from the more unsaturated milk was less oxidatively stable during storage (at 4 °C, followed for 11 d) than buttermilk from the more saturated milk, as monitored both by primary lipid oxidation products (lipid hydroperoxides) and by the secondary lipid oxidation product, hexanal. Fat-soluble antioxidants, β-carotene and α-tocopherol, analysed by HPLC, were not consumed during storage for either of the two types of buttermilk. In contrast, the antioxidative capacity of the serum phase decreased during storage as evaluated in a radical scavenging assay based on the semi-stable water-soluble radical nitrosodisulphonate (Fremy's salt). The time course for the decrease in water-soluble antioxidants was very similar for the two types of buttermilk suggesting that oxidation is initiated in the serum phase independently of fatty acid composition.