Magnetic fluids, as smart nanomaterials, have been successfully used in sealing applications and other fields. However, the temperature of magnetic fluids in the sealing gap is a key factor affecting sealing performances, limiting their application in high-speed sealing fields. Since obtaining a direct measurement of the magnetic fluid’s temperature is difficult, due to the small clearance, accurately calculating the maximum temperature of the magnetic fluid layer in high-speed seals is crucial. Herein, a mathematical model for calculating the maximum temperature of the magnetic fluid layer was established, by using a reasonable simplification of high-speed sealing conditions, and the calculation formula was modified by studying the rheological properties of the diester-based magnetic fluid. The results suggest that the calculation of the maximum temperature is influenced by viscous dissipation, and both are related to the rheological characteristics of magnetic fluids. When the influence of rheological properties is ignored, the calculation results are not accurate for higher-velocity seals, but the calculation model applies to lower-velocity seals. When the influence of rheological properties is considered, the calculation results obtained by the corrected formula are more accurate, and they are applicable to both lower- and higher-velocity seals. This work can help us more accurately and conveniently estimate the maximum temperature of magnetic fluids in high-speed seal applications, which is of theoretical and practical research significance for determining sealing performances and thermal designs.