The feasibility of pressure-retarded osmosis (PRO) for power generation was evaluated with consideration of the energy inputs and losses in the process. The effects of the concentration polarization, reverse salt diffusion, and external resistance at the membrane porous layer were quantified, for the first time, along the membrane module to determine their contributions to the energy loss in the PRO process. Concentration polarization was responsible for up to 40% of the energy loss during the PRO process. However, increasing the PRO membrane modules from 1 to 4 resulted in a variable increase of the energy output depending on the salinity gradient. The energy requirements for draw and feed solution pretreatment were estimated to be over 38% of the total energy inputs. Results showed that coupling seawater (SW) with river water (RW) was unable to generate sufficient energy to compensate for the energy inputs and losses during the PRO process. With 0.39 kwh/m3 maximum specific energy in the PRO process, the energy yield of reverse osmosis brine (ROB)-wastewater (WW) salinity gradient was slightly greater than the total energy inputs, although using Dead Sea-SW/ROB salinity gradient was more promising. Overall, the primary current limitation is the lack of suitable PRO membranes that can withstand a high hydraulic pressure.