Fat-soluble vitamins transfer poorly across the diffuse epitheliochorial placenta of the mare, so the neonatal foal is dependent on its supply via colostrum. Concentrations of retinol (RT), β-carotene (BC), and β-tocopherol (AT) were assayed in samples of plasma, colostrum and milk from nine mares of mixed light breeding and their foals at parturition and days 1, 2, and 4 postpartum. Samples were analyzed simultaneously for RT, BC, and AT using a new, less time-consuming reverse phase high-pressure liquid chromatography (HPLC) method. Feeds were analyzed, and mean daily intakes calculated. Mare plasma RT increased from day 1 to day 4 (P = .033), and plasma AT declined linearly from day 0 to day 4 (P = .006). Colostrum concentrations of all vitamins increased from parturition to day 1, and then declined rapidly over the 4 days. Foal plasma BC increased from parturition to day 4 (P = .080), and plasma AT increased from parturition to day 2 (P < .001), and 4 (P = .060). These observations suggest that three times the current recommendation of vitamin A is sufficient for pregnant mares, in keeping with previous studies of growth, pregnancy, and lactation. In contrast, the linear decrease in mare plasma AT suggests that 1.2 times the current recommendation of vitamin E may be less than optimal for pregnant mares.