The effect of the imbibition temperature on the operational performance of the extraction stage is not fully clarified. Additionally, many criteria referred to on this issue are contradictory, inconclusive, or not supported by industrial data. Due to such limitations in knowledge, this research aims to analyze the influence of imbibition temperature on: i) temperature of the milling train juices, ii) moisture and sucrose composition of bagasse, and iii) starch composition of milling train juices. The results indicate that the imbibition temperature has no influence on the mixed juice temperature and does not allow the minimum recommended temperature for asepsis (65 °C) in all juices of the milling units to be reached. Moreover, from the experiments performed, it is established that bagasse moisture increases by 1.6–1.8 units, and sucrose extraction reduces by 12% when the imbibition temperature varies from 85 °C to 40 °C. The starch composition of the mixed juice increases by 15% when the imbibition temperature rises from 40 °C to 85 °C. The research provides new statistical evidence for a better understanding and operational control of the effect of imbibition temperature.