The oil recovery improvement by low-salinity waterflooding (LSWF) must be achieved with minimal formation damage. The asphaltic fractions of crude oil can be destabilized when in contact with incompatible injection brines resulting in organic formation damage. Due to the lack of fundamental understanding about the potential effect of rock presence on this phenomenon, in this study, the simultaneous effect of brine salinity and calcite rock presence on asphaltene instability in an emulsified system was investigated by developing a new experimental protocol. In this method, calcite rock is dispersed in different brines with total salinity ranging from 0−189,365 ppm and then mixed with oil to find the rock and brine effect on asphaltene and emulsion stability. UV−vis spectroscopy is performed on the supernatant oil phase before and after contact with brine to assess quantitatively asphaltene instability. Oil/water IFT measurements before and after contact and microscopic analysis of the water droplet size were conducted to evaluate the water/oil interface behavior. Besides, the zeta-potential for suspended calcite particles in different brines was measured to evaluate the electrokinetics of rock-oil interaction. The results show increased asphaltene instability when oil makes contact with the brine in an emulsified system. This highly depends on the brine type, and the maximum precipitation occurred at an intermediate salinity with two-times diluted seawater. While the trend of before-contact IFT and the water-in-oil emulsion size at different salinities is consistent, the UV−vis absorbance of bulk oil shows a reverse tendency with the after-contact IFT. Asphaltene deposition generally increased when solid calcite particles were present in the emulsified system, and at higher salinities, more asphaltene adsorption on the rock surface occurred due to a larger attractive force between rock and asphaltene. This trend is consistent with the results of the zeta-potential measurements. These novel findings can help avoid unrepresentative results regarding organic damage in LSWF.