An efficient cooling system is essential for the electronic packaging such as a high-luminance LED lighting. A special heat transport technology, Pulsating Heat Pipe (PHP), can be applied to the cooling of LED lighting. In this paper, the operational characteristics of the PHP in the imposed thermal boundary conditions of LED lighting were experimentally investigated. The experimental PHP was made of copper tubes of internal diameter of 2.1 mm. The working fluids of ethanol, FC-72, water, acetone and R-123 were chosen for comparison. The results showed that an optimum range of charging ratio exists for high cooling performance; 50% for most of the fluids. Among the five working fluids, water showed the highest heat transfer rate of 260 W. Two distinguished characteristics of pulsating direction were identified. It is also identified that high vapor pressure gradient is one of key parameters for better heat transfer performance.