Assessing the metabolic profile based on the concept that the laboratory measurement of certain circulating components is a tool to evaluate metabolic status of dairy cows. Veterinarian also can evaluate the energy input-output relationships by assessing the metabolic profile to prevent and control of negative energy balance, metabolic disorders and nutritional insufficiencies. In the present study, 25 multiparous Holstein dairy cows were divided to 5 equal groups containing early, mid and late lactation, and far-off and close-up dry. Blood samples were collected from all cows through jugular venipuncture and sera were evaluated for glucose, insulin, β-hydroxybutyric acid (BHBA), non-esterified fatty acid (NEFA), cholesterol, triglyceride (TG), high, low and very low density lipoproteins (HDL, LDL and VLDL). Insulin levels in mid lactation and close-up dry cows were significantly higher than other groups (P<0.05) and the lowest insulin concentration was detected in faroff dry group. Serum concentrations of NEFA and BHBA in early and mid-lactation and close-up dry cows were significantly higher than late lactation and far-off dry animals (P<0.05). Baseline levels of cholesterol in mid and late lactation were significantly higher than other groups. The level of LDL in mid lactation cows was higher than others significantly, and its value in far-off dry cows was significantly lower than other group (P<0.05). It may be concluded that the detected changes among different groups induce commonly by negative energy balance, lactogenesis and fetal growth in each state. The presented metabolic profile can be considered as a tool to assess the energy balance in dairy cows at different physiologic states. It can be used to evaluate the metabolic situations of herd and manage the metabolic and production disorders.