The aim of this study was to assess the effect of 2 different megadoses of retinyl palmitate on the level of retinol in the breast milk of healthy women. In total, 199 women were randomly allocated to 3 groups and supplemented in the postpartum period with a single retinyl palmitate dose of 200 000 IU (S1), a double dose of 200 000 IU 24 hours apart (S2), or no supplementation (C). Retinol content of colostrum and mature milk at 4 weeks was determined by high-performance liquid chromatography (HPLC). For colostrum, no significant difference was found between the groups (P = .965). The retinol content in mature milk differed between group C and groups S1 and S2 (P < .05). The double dose of vitamin A did not significantly increase the retinol content of milk at 4 weeks postpartum in comparison to a single dose; however, future research is needed to determine the optimal timing of the second dose of vitamin A.