Due to the complexity of the automobile manufacturing process, some flexible and delicate assembly work relies on manual operations. However, high-frequency and high-load repetitive operations make assembly workers prone to physical fatigue. This study proposes a method for evaluating human physical fatigue for the manual assembly of automobiles with methods: NIOSH (National Institute for Occupational Safety and Health), OWAS (Ovako Working Posture Analysis System) and RULA (Rapid Upper Limb Assessment). The cerebral oxygenation signal is selected as an objective physiological index reflecting the human fatigue level to verify the proposed physical fatigue evaluation method. Taking auto seat assembly and automobile manual assembly as an example, 18 group experiments were carried out with the ARE platform (Augmented Reality-based Ergonomic Platform). Furthermore, predictions of metabolic energy expenditure were performed for experiments in Tecnomatix Jack. Finally, it is concluded that the proposed physical fatigue evaluation method can reflect the human physical fatigue level and is more accurate than the evaluation of metabolic energy consumption in Tecnomatix Jack because of the immersion that comes with the AR devices and the precision that comes with motion capture devices.