The present study was designed to determine the effect of barn or cycle of grazing on changes of biochemical metabolites in prepartum and changes of milk composition in postpartum of dairy cows. For this purpose, a total of sixteen 25 months old Holstein primiparous dairy cows were allocated in two groups (n=8) with an average body weight of 571.61 ± 35.30 kg (Barn) and 578.10 ± 39.20 kg (Grazing). The study was conducted from June 2018 to October 2018. Results revealed that barn raised dairy cows had a higher increase in their serum albumin and calcium level on day 14 prepartum. However, the level of palmitic acid, saturated fatty acid increased significantly, and the level of fat, oleic acid, γ-linoleic acid, arachidonic acid and unsaturated fatty acids decreased significantly in barn raised dairy cow’s milk on day 14 postpartum. There were no significant differences observed with respect to all other biochemical metabolites, fatty acids and minerals between barn raised and cycle grazing dairy cows during prepartum and postpartum. Our study results could serve to a better understanding of barn raised cow with respect to changes of biochemical metabolites in prepartum and changes of milk composition, fatty acids and minerals content in grazing dairy cows in postpartum for estimating their physiological status.