The Calcium ion Ca2+ plays a critical role as an initiator and preserving agent of the cross-bridge cycle in the force generation of skeletal muscle. A new multi-scale chemo-mechanical model is presented in order to analyze the role of Ca2+ in muscle fatigue and to predict fatigue behavior. To this end, a cross-bridge kinematic model was incorporated in a continuum based mechanical model, considering a thermodynamic compatible framework. The contractile velocity and the generated active force were directly related to the force-bearing states that were considered for the cross-bridge cycle. In order to determine the values of the model parameters, the output results of an isometric simulation were initially fitted with experimental data obtained for rabbit Extensor Digitorum Longus muscle. Furthermore, a simulated force-velocity curve under concentric contractions was compared with reported experimental results. Finally, by varying the Ca2+ concentration level and its kinetics in the tissue, the model was able to predict the evolution of the active force of an experimental fatigue protocol. The good agreement observed between the simulated results and the experimental outcomes proves the ability of the model to reproduce the fatigue behavior and its applicability for more detailed multidisciplinary investigations related to chemical conditions in muscle performance. © 2018 Elsevier Ltd