Bile acids are traditionally known for their role in lipid digestion and recently bile acids have been shown to have potential endocrine activity. Pregnancy state elevates bile acid concentration in humans, but the effects of pregnancy on bile acid concentrations have not been studied in cattle. Our objective was to report the bile acid profile during the transition period and to determine if bile acid concentrations are associated with energy balance, glucose and insulin. Multiparous Holstein cows (n = 48) were characterized for plasma total bile acid concentration, energy balance, nonesterified fatty acids (NEFA), BHB, glucose and insulin at 5 time points: 3 wk prepartum (wk −3), 2 wk prepartum (wk −2), week of calving (Wk0), first week postpartum (wk +1) and second week postpartum (wk+2). Blood was collected at approximately 0900 h and fresh TMR was given at 0800 h daily. Cows were individually fed in tie-stalls with daily feed offered and refused weighted to determine feed intake. Average daily energy balance was calculated based on energy intake and energy requirements for maintenance, pregnancy or lactation for each of the time periods. Data were analyzed as repeated measures using PROC MIXED of SAS. Total bile acid concentration was significantly lower during pregnancy and increased postpartum (in µmol/L, wk −3 = 15.7 ± 1.3, wk −2 = 14.9 ± 1.3, wk 0 = 38.7 ± 2.6, wk +1 = 65.1 ± 5.0, wk +2 = 75.2 ± 9.1; P < 0.001). Plasma total bile acid concentration was positively associated with NEFA (P < 0.001) and BHB (P = 0.009); and negatively associated with energy balance (P < 0.001) and insulin (P < 0.001); but not associated with glucose (P = 0.19). Glucose to insulin ratio was also higher in cows with increased plasma total bile acid (P < 0.001). Taken together, increased plasma total bile acid was associated with poorer energy balance and higher glucose to insulin ratio during the transition period but further studies are warranted. This work was supported by the USDA National Institute of Food and Agriculture, Research Animal Health project NYCV-480867.