The effect of temperature and brine composition on Crude-Oil-Rock-Brine (CORB) interactions during controlled salinity brine and biosurfactant injection process were investigated and reported in this study. Comprehensive core flooding experiments were carried out using formation brine, controlled salinity brine and combined controlled salinity biosurfactant brine injection in carbonate core samples at 23 °C and 70 °C. Effluent analyses and oil recovery from each flooding experiment were used to interpret the temperature and brine composition effect on the CORB interactions. The results of this study show that increased temperature makes no significant impact on CORB interactions during high salinity brine flooding. Increased temperature however, enhances CORB interactions through increased reactivity during controlled salinity brine and controlled salinity biosurfactant brine flooding thereby, resulting in increased oil production. Furthermore, the results of this study demonstrated the enhanced oil recovery potential of combined controlled salinity brine and biosurfactant application in condition relevant to hydrocarbon reservoirs.