The effects of vitamin AD3E (V-AD3E) solution on the milk protein expression of periparturient dairy cows were determined using proteomic analysis. Milk Protein from d 1 and d 21 after calving were separated and identified by two-dimensional electrophoresis combined with high performance liquid chromatography tandem ion trap spectrometry (HPLC/MS/MS). IgG and albumin in colostrum from dairy cows injected with V-AD3E solution were up-regulated compared with not injected ones. No changes of milk protein composition were observed from d 21 postpartum between injection and non-injection. Injecting V-AD3E solution into the periparturient dairy cows prior to calving up-regulated IgG and albumin levels in colostrum,which, besides transferring higher levels immune constituents to the offspring, fed the colostrum, also promotes development and digestive tract protection in the neonate, and defends the cows who are in immune suppression period against the invasion of pathogenic bacteria,fungi and virus.