Nitrogen is an alloying element that can significantly improve the yield strength of steel by means of interstitial solid solution strengthening. The effects of quenching temperature on the evolution of second phase and grain size were investigated, as well as tensile properties, impact energy and hardness of a new nitrogen-alloyed Fe-Cr-Mo-V-N steel. The results show that with the increase of quenching temperature, the number and size of the second phase decrease as the high temperature promotes the dissolution and refinement of the second phase. For instance, the number of second phase M 23 C 6 particles decreases gradually with temperature increasing. When the quenching temperature reaches 1050 °C, the grain is refined. Under the same tempering treatment, with the increase of quenching temperature, the tensile properties, impact energy and hardness increase first and then decrease. When the quenching temperature is 1050 °C and the tempering temperature is 180 °C, the obtained mechanical properties are the best. Under this condition, the tensile strength at room temperature is 2110 MPa, the yield strength is 1620 MPa, and the impact energy is 29.1 J, respectively.