Bones form protective and load-bearing framework of the body. Therefore, their structural integrity is vital for the quality of life. Unfortunately, bones can only sustain a load until a certain limit, beyond which they fail. Therefore, it is essential to study their mechanical and fracture behaviours in order to get an in-depth understanding of the origins of its fracture resistance that, in turn, can assist diagnosis and prevention of bone's trauma. This can be achieved by studying mechanical properties of bone, such as its fracture toughness. Generally, most of bone fractures occur for long bones that consist mostly of cortical bone. Therefore, in this study, only a cortical bone tissue was studied. Since this tissue has an anisotropic behaviour and possesses hierarchical and complex structure, in this paper, an experimental analysis for the fracture toughness of cortical bone tissue is presented in terms of J-integral. The data was obtained using single-edge-notch bending (SENB) cortical specimens of bone tested in a three-point bending setup. Variability of values of fracture toughness was investigated by testing specimens cut from different cortex positions of bovine femur called anterior, posterior, medial, and lateral. In addition, anisotropy ratios of fracture toughness were considered by examining specimens cut from three different orientations: longitudinal, transverse and radial. Moreover, in order to link cortical bone fracture mechanisms with its underlying microstructure, fracture surfaces of specimens from different cortices and along different orientations were studied. Experimental results of this study provide a clear understanding of both variability and anisotropy of cortical bone tissue with regard to its fracture toughness.