Passive mechanical properties in three different zones (distal, proximal, and medial) of biceps brachii immobilized in short position and its free contralateral were investigated. For that, in vitro equibiaxial relaxation tests were performed with samples harvested from skeletal muscles of immobilized rats during one or two weeks. From data obtained in two plane axes of loading, a viscohyperelastic anisotropic model described by a strain energy function coupled with second order Maxwell's model, was used to identify the material parameters. It has been shown that the zone influences the material parameters of the hyperelasticity behavior while the immobilization acts rather on the viscoelasticity response. Based on measurements of histological parameters from flexor carpi ulnaris muscles, we observed that immobilization produced contractile tissue atrophy and connective tissue thickening. The muscle atrophy caused by immobilization could lead to a more linear mechanical behavior along the axis of the muscle fibers. Furthermore, fibrosis, quantified by histological analysis on flexor carpi ulnaris muscles, could result in highly non-linear behavior along the other axis. These structural changes also contribute to an increase in relaxation following immobilization along both axes (+11.7% and +15.5% on average with p < 0.001 and p < 0.01 respectively for each axis). Statement of significanceTo our best knowledge, this work is the first to investigate the anisotropic, viscohyperelastic and heterogeneous properties within a passive skeletal muscle immobilized in a short position. For that, from a rat model, equibiaxial relaxation tests were performed and material parameters were found from a strain energy function coupled with a Maxwell's model. We showed that excising zone of samples greatly influences the parameters of the hyperelastic behavior while the immobilization influences the viscoelasticity response instead. Furthermore, histological analysis permitted to quantify fibrosis in the skeletal muscle and bear out, at least in part, changes in relaxation behavior following immobilization.