This work aims to determine how the temperature gradient orientation affects the heat exchange between two superposed fluid layers separated by zero wall thickness. The finite volume method (FVM) has been developed to solve the governing equations of both fluid layers. To achieve the coupling between the two layers, the heat flow continuity with the no-slip condition at the interface was adopted. The lower part of the space is filled with a nanofluid while the upper part is filled with a pure fluid layer. We have explored two cases of temperature gradient orientation: parallel gradient to gravity forces of our system and perpendicular gradient to gravity forces. We took a set of parameters, Ri and ϕ, to see their influence on the thermal and hydrodynamic fields as well as the heat exchange rate between the two layers. The main applications of this study related to biological systems such as the cytoplasm and the nucleoplasm are phase-separated solutions, which can be useful as models for membranelles organelles and can serve as a cooling system application using heat exchange. The Richardson number and the volume of nanosolid particles have a big impact on the rate of change of heat transmission. When a thermal gradient is perpendicular to gravity forces, total heat transmission improves with increasing solid volume percentage, but when the thermal gradient is parallel to gravity forces, overall heat transfer decreases significantly.