The effects of thermal aging and step cooling embrittlement on the impact toughness of a reactor pressure vessel steel SA533B quenched and tempered (QT) with and without post-weld heat treatment (PWHT) have been studied. Charpy impact testings were conducted on the aged plates at 350°C for 5000 h to evaluate whether the embrittlement was induced by step cooling heat treatment. The results show that thermal aging increases the ductile–brittle transition temperature in both QT and PWHT states but dramatically decreases the upper shelf energy in QT state and has less effects on the PWHT state. By comparing the correlation between thermal aging embrittlement and step cooling embrittlement for both QT and PWHT states in steel, it is found that the step cooling heat treatment can obviously promote further embrittlement of the base metal in QT state but has little influence on the impact toughness in PWHT and thermal aged state. Further analysis indicates that the step cooling heat treatment cannot promote steel embrittlement at some heat treatment states. Finally, a new method is proposed to evaluate the degree of step cooling embrittlement of the pressure vessel steel.