A statistical re-assessment of aggregated individual cow data was conducted to examine trends in fat-to-protein ratio in milk (FPR), and relationships between FPR and energy balance (EB, MJ of ME/day) in Holstein-Friesian dairy cows of different parities, and at different stages of lactation. The data were collected from 27 long-term production trials conducted between 1996 and 2016 at the Agri-Food and Biosciences Institute (AFBI) in Hillsborough, Northern Ireland. In total, 1321 lactations (1 to 20 weeks in milk; WIM), derived from 840 individual cows fed mainly grass silage-based diets, were included in the analysis. The energy balance was calculated daily and then averaged weekly for statistical analyses. Data were further split in 4 wk. intervals, namely, 1–4, 5–8, 9–12, 13–16, and 17–20 WIM, and both partial correlations and linear regressions (mixed models) established between the mean FPR and EB during these periods. Three FPR score categories (‘Low’ FPR, <1.0; ‘Normal’ FPR, 1.0–1.5; ‘High’ FPR, >1.5) were adopted and the performance and EB indicators within each category were compared. As expected, multiparous cows experienced a greater negative EB compared to primiparous cows, due to their higher milk production relative to DMI. Relatively minor differences in milk fat and protein content resulted in large differences in FPR curves. Second lactation cows displayed the lowest weekly FPR, and this trend was aligned with smaller BW losses and lower concentrations of non-esterified fatty acids (NEFA) until at least 8 WIM. Partial correlations between FPR and EB were negative, and ‘greatest’ in early lactation (1–4 WIM; r = −0.38 on average), and gradually decreased as lactation progressed across all parities (17–20 WIM; r = −0.14 on average). With increasing parity, daily EB values tended to become more negative per unit of FPR. In primiparous cows, regression slopes between FPR and EB differed between 1–4 and 5–8 WIM (−54.6 vs. −47.5 MJ of ME/day), while differences in second lactation cows tended towards significance (−57.2 vs. −64.4 MJ of ME/day). Irrespective of the lactation number, after 9–12 WIM, there was a consistent trend for the slope of the linear relationships between FPR and EB to decrease as lactation progressed, with this likely reflecting the decreasing milk nutrient demands of the growing calf. The incidence of ‘High’ FPR scores was greatest during 1–4 WIM, and decreased as lactation progressed. ‘High’ FPR scores were associated with increased energy-corrected milk (ECM) yields across all parities and stages of lactation, and with smaller BW gains and increasing concentrations (log transformed) of blood metabolites (non-esterified fatty acid, NEFA; beta-hydroxybutyrate, BHB) until 8 WIM. Results from the present study highlight the strong relationships between FPR in milk, physiological changes, and EB profiles during early lactation. However, while FPR can provide an indication of EB at a herd level, the large cow-to-cow variation indicates that FPR cannot be used as a robust indicator of EB at an individual cow level.