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Accurately realizing the use of Charpy impact test to measure fracture toughness and determine reference temperature of low alloyed steel with ferritic microstructure has important practical engineering significance for the structural integrity assessment of reactor pressure vessels. In this paper, a methodology for measuring fracture toughness and determining reference temperature for RPV steels by Charpy impact test was proposed. The method referred to the ASTM E1820 and E1921 standard. By analyzing the physical meaning of the Charpy impact specimen’s fracture, load-time curve and size parameters, the fracture toughness of the material was directly calculated according to the linear elastic fracture mechanics. The reference temperature was calculated using the multi-temperature procedure formula. The reference temperature and fracture toughness were corrected according to two factors, such as the difference in fracture toughness caused by the difference in the loading rate of the stress intensity factor and the specimen shape and size between Charpy impact test and compact tension test. And the validity of the fracture toughness and reference temperature was evaluated. The corrected two are basically consistent with the data obtained by the compact tension test in accordance with ASTM E1820 and E1921 standards. This method may be used to solve the problem that the nuclear power plant needs to use the impact test data to evaluate the fracture toughness and reference temperature of the reactor pressure vessel in the structural integrity assessment of the reactor pressure vessel operation and life extension process.