Abstract The mathematical model of the compartmentalised energy transfer system in cardiac myocytes, which includes mitochondrial synthesis of ATP by ATP-synthase, phosphocreatine production in the coupled mitochondrial creatine kinase reaction, the myofibrillar and cytoplasmic creatine kinase reactions, ATP utilisation by actomyosin ATPase during contraction cycle, and diffusional exchange of metabolites between different compartments, was used to calculate creatine kinase reaction rates (fluxes) in different cellular compartments at a workload corresponding to the rate of oxygen consumption of 46 μg-atom O 2* min −1* (g wet mass) −1. The results of calculations showed that at this high workload all creatine kinase isoenzymes function most of their time in the cardiac cycle in the steady state far from equilibrium. This mathematical modelling shows that the validity of assumption of creatine kinase equilibrium is limited only to the diastolic phase of the contraction cycle in the working cardiac cells and only to the cytoplasmic compartment. In the systolic phase, due to rapid release of ADP at increased workloads, all creatine kinase isoenzymes are rapidly shifted out of the equilibrium. Cytoplasmic ADP concentration may increase up to 9 times in the systolic phase of the cardiac cycle, correspondingly changing all ADP-dependent parameters. Mitochondrial creatine kinase functions permanently in “metastable” steady state (Jurgen Daut, Biochim. Biophys. Acta 895,41–62, 1987). It may be proposed that a more precise, in comparison to the equlibrium concept, way of calculating steady state cytoplasmic ADP concentrations at increased workloads is to use kinetic equations and mathematical models of energy metabolism.