The effect of tempering temperature on the impact toughness of 0.3 mass% carbon martensitic steels with prior austenite grain (PAG) size of about 6 μm and 30 μm were investigated. Instrumented Charpy impact test (ICIT) method was used to evaluate the impact toughness. The tempering temperature of 723K gives the largest difference in the Charpy impact energy at room temperature (RT) between the specimens with two different PAG sizes. Investigation of the test temperature dependence of Charpy impact energy in the 723K tempered steels shows a steep transition at around 200 K for the 6 μm PAG specimen, while it shows a continuous slow transition in a wide range of temperature for the 60 μm PAG specimen. ICIT waveform analysis shows that the fracture propagation energy in stead of the fracture initiation energy mainly controls the temperature dependence of the impact energy. The carbide size distribution in these two specimens was investigated by SEM and TEM. The 60 μm PAG specimen shows the distribution of coarser carbides than does the 6 μm PAG specimen, which seems to be the main reason for the observed difference between them in the Charpy impact energy and the other properties of impact fracture.