The aim of the present study was to determine the metabolic profile of Holstein-Friesian cows in the postpartum period, as well as the effect of season on metabolic profile. The postpartum period is essential in the reproductive life of high yielding dairy cows because of its impact on future gravidity. This study included 60 cows up to 15 days after parturition, aged 2-8 years (the largest number of cows was between 3 and 5 years old) with no apparent clinical problems. Cows were sampled in summer season (n=30) and winter season (n=30). Parameters of metabolic profile were determined as follows: glucose, albumin, total protein, cholesterol, bilirubin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), urea, calcium, phosphorus and magnesium. Statistical differences were considered significant at the p<0.05. Present research showed that all investigated parameters were within a reference range for cattle. Impact of season sampling was determined for glucose, albumin, total protein, cholesterol and phosphorus, while bilirubin, calcium, magnesium, urea as well as activities of ALT, AST and LDH were unaffected by the season of sampling. In conclusion, metabolic status is affected by the season and examination during the postpartum period can provide valuable information of cows' health status, in order to diagnose and moreover prevent postpartum diseases.