Measurement of fracture toughness is one of the quality control parameter in rail manufacturing process. Fracture toughness value is needed for designing the rail lines, analysis of defects and other common prevalent works in mechanical engineering. The goal of this research is to introduce a relation for measuring fracture toughness of rail materials with grade R260 by Charpy V-notch number. At first the fracture toughness of rail material has been determined by 3-point bending technique according to ASTM E399 in ambient temperature. The fracture energies have been measured by Charpy impact test and it has been illustrated that fracture energies have no significant change in the tests temperature limit. Relations between the chemical analysis and mechanical properties have been studied and compared with results of the other researcher. Uniaxial tension test, analysis of chemical composite, metallography and hardness test have been carried out to better study of the process. Finally a relation to connect the Charpy V-notch number and fracture toughness has been introduced and the fracture toughness of the rails material over the impact test temperature limit has been calculated. Good agreement between calculated results and result of 3-pint bending test indicate appropriate accuracy of the introduced equation.