Pulsating heat pipes are high efficiency heat transfer components having a great potential application in the field of electronic cooling and space applications. In this investigation, an experiment was conducted to study the influence of working fluid thermophysical properties on the thermal performance of flat-plate closed loop pulsating heat pipes (FCLPHP). The analysis of the forces acting on the liquid-vapor mixture shows that the surface tension, viscosity and latent heat of vaporization have important impact on the thermal performance of FCLPHP. The overall thermal resistance decreases with the decrease in surface tension, viscosity and latent heat of vaporization, leading to the heat transfer improvement of FCLPHP. An experimental correlation was developed to describe the relationship among the relative overall thermal resistance and the relative thermophysical properties. With the correlation, a sensitivity analysis was made. The results show that latent heat of vaporization is the prior factor to the all others to impact the thermal performance of FCLPHP.