Availability of biaxial mechanical data for heart myocardia remains high in demand for the development of accurate and detailed computational models. The aim of this study is to study the regional difference of wall mechanics using rat heart in the left ventricle (LV), septal wall (STW) and right ventricle (RV). This was achieved by conducting a biaxial test on three rat heart myocardia (i.e LV, RV and STW). Fung, Choi-Vito, Polynomial (Anistropic), Four-Fiber family, Holzapfel (2000) and Holzapfel (2005) hyperelastic models were selected and fitted on the bixial data of the LV, RV and STW. The best hyperelastic model was the selected based on evaluation index (EI) determined from the coefficient of determination (R2). All the six models were then compared in all three rat heart myocardia. The results show that the Polynomial (Anisotropic) model outperforms the other five models in all myocardial tissues with EI’s above 90 % goodness of fit. The Four-fiber-family and the two Holzapfel models perform equally in the LV and STW myocardial tissue between 50 and 70 % goodness of fit. The Fung and Choi-Vito models yielded poor goodness of fit in the LV and STW myocardial tissues. Parameter fitting is useful method in advancing reliable data to be used in the development of accurate computational models.