The effect of oral vitamin E supplementation during the dry period on serum biomarkers of oxidative status was evaluated in an observational field study. Ninety-eight cows were selected from 26 herds in the Netherlands. On 15 farms dry cows were supplemented with a dry-cow mineral mix containing 1000 IU vitamin E (vitamin E+ group, n = 52), and on 11 other farms dry cows did not receive the supplement (control group, n = 48). A blood sample was taken from all cows between 0 and 28 days before calving. Serum levels of alpha-tocopherol, cholesterol, triacylglycerol, superoxide dismutase (SOD), glutathione peroxidase (GSHpx), uric acid, albumin, protein sulphydryl oxidation level, reactive oxygen metabolites (ROM), iron, the ferric reducing ability (FRAP), and malondialdehyde (MDA) were measured. Alpha-tocopherol levels were significantly higher in the vitamin E+ group, as was the alpha-tocopherol:cholesterol ratio. MDA levels were significantly lower in the vitamin E+ group, suggesting that levels of lipid peroxidation were lower with vitamin E supplementation. In conclusion, vitamin E supplementation increases alpha-tocopherol and lowers MDA serum levels.