Natural circulation loop (NCL) is simple and reliable due to the absence of moving components and is preferred in applications where safety is of foremost concern, such as nuclear power plants and high-pressure thermal power plants. In the present study, optimum operating conditions based on the maximum heat transfer rate in NCLs have been obtained for subcritical as well as supercritical fluids. In recent years, there is a growing interest in the use of carbon dioxide (CO2) as loop fluid in NCLs for a variety of heat transfer applications due to its excellent thermophysical environmentally benign properties. In the present study, three-dimensional (3D) computational fluid dynamics (CFD) analysis of a CO2-based NCL with isothermal source and sink has been carried out. Results show that the heat transfer rate is much higher in the case of supercritical phase (if operated near pseudocritical region) than the subcritical phase. In the subcritical option, higher heat transfer rate is obtained in the case of liquid operated near saturation condition. Correlations for optimum operating condition are obtained for a supercritical CO2-based NCL in terms of reduced temperature and reduced pressure so that they can be employed for a wide variety of fluids operating in supercritical region. Correlations are also validated with different loop fluids. These results are expected to help design superior optimal NCLs for critical applications.