Ceruloplasmin (Cp) plays an important role in copper transport and iron metabolism, as well as Cp is also an indicator for the health status of dairy cows. The present study reports the validation of an automated assay to assess the plasma Cp in dairy cows. Plasma Cp levels were determined in 40 Holstein cows and intra- and inter-assay precision, accuracy, detection limit, and clinical validation of the assay were determined. Intra- and inter-assay coefficients of variation were < 2% and < 7%, respectively. The results were linear when serial sample dilutions were tested (r = 0.999, P < 0.001). The detection limit was lower than what could be measured in plasma from healthy cows. Increased plasma Cp levels were found in cows with inflammatory diseases. The method validated in this study is precise, simple, and fast and can be easily adapted to biochemical automated analysers. Furthermore, the promising results obtained with this protein will contribute to a wider use of Cp determination in bovine practice.