Mobilized fatty acids (FA) provide an energy source to the liver during negative energy balance but can also be stored as liver lipids. The objectives were to determine the change in FA profile across the transition period and the correlations between circulating FA profile changes, blood concentration, and liver lipid profile in Holstein cows. Blood samples were taken at −28, +1, +3, +14, and +28 (n = 36, 27, 25, 38, and 25, respectively) and liver biopsies were taken at +1, +14, and +28 (n = 27) days relative to calving (DRTC). Liver and plasma FA were determined by acid methylated and gas chromatography. To calculate delta plasma values, the proportion of each FA at −28 was subtracted from the proportion on each day postpartum. Data were analyzed with PROC MIXED and CORR (SAS 9.4). The mixed model included the fixed effect of DRTC and the random effect of cow using repeated measures and data expressed as mean ± SE. Proportionally, C16:0 and C18:1 were increased (P < 0.01) postpartum and were greatest at +1 (4.0 ± 0.4 and 7.5 ± 0.6%) and +3 DRTC (4.0 ± 0.4% and 7.5 ± 0.6% increase for C16:0 and C18:1). Plasma FA concentration and delta plasma concentrations at +1 DRTC were correlated (P < 0.01) for C16:0 (r = 0.57) and C18:1 (r = 0.74). These data support that C16:0 and C18:1 are the primary mobilized FA from adipose tissue. Relative to −28 DRTC, the proportion of C18:2 was decreased (P < 0.01) at +1 (−6.3 ± 1.1%) and +3 DRTC (−6.0 ± 1.1%) yet greater at +14 (2.7 ± 1.1%) and +28 DRTC (4.5 ± 1.1%) suggesting contributions beyond adipose mobilization. The proportion of C18:0 was decreased (P < 0.01) postpartum (−2.3 ± 0.5%) and not correlated (P > 0.1) to total plasma FA. The correlation between plasma and liver profiles differed by fatty acid. Plasma and liver C18:0 was not correlated (P > 0.1) on +1, +14 and +28 DRTC (r = 0.20, 0.10, −0.23). Correlations were greater between plasma and liver for C16:0 (r = 0.59, 0.35, 0.33), C18:1 (r = 0.79, 0.60, 0.60), and C18:2 (r = 0.5, 0.49, 0.46) at +1, +14 and +28 DRTC. Taken together, these data suggest that circulating FA during negative energy balance reflect adipose mobilization patterns and preferential metabolic fates in hepatic tissue that should be further examined.