Radiographic assessment tools such as the cortical step sign (CSS) or the diameter difference sign (DDS) aim to identify clinically relevant rotational malalignment after long bone fracture fixation. We aimed to analyze the effect of rotational malalignment on CSS and DDS parameters in a subtrochanteric fracture model and to construct a prognostic model to identify clinically relevant rotational malalignment. A subtrochanteric transverse osteotomy was set in human femora. Rotation was set stepwise from 0° to 30° in internal and external rotation. Images were obtained using a C‐arm and transferred for measuring the medial cortical thickness (MCT), lateral cortical thickness (LCT), femoral diameter (FD) in AP and the anterior cortical thickness (ACT) as well as the posterior cortical thickness (PCT) and the FD of the proximal and the distal main fragment. There were significant differences between the various levels of rotation for each of the absolute values of the evaluated variables. MCT, PCT and FD (AP & lat.) were the most affected parameters. In internal rotation, the MCT, PCT and the FD were the most affected variables. The parameters with the highest correlation with femoral rotation were ACT, PCT and FD. A model combining ACT, LCT, PCT and FD AP was most suitable model in identifying rotational malalignment. The best prediction of clinically relevant rotational malalignment was obtained with the FD and the PCT. The CSS and the DDS are promising tools for detecting rotational deformities of the proximal femur and should be used intra‐ and postoperatively. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res