Objective To investigate an ideal method of treating bovine pericardial. Methods The changes of biomechanical and ultrastructural properties of the bovine pericardial affer different treatments were studied by the shrinkage temperature analysis and uniaxial load test. Results After the treatment, the thickness and stiffness of pericardium were increased, whereas the plasticity and flexibility were decreased, which became similar to the biomechanical properties of bioprosthesis. The modulus of elasticity of 2,3-butanediol post-treatment group is higher than that of glutaraldehyde-fixed group (8.75±1.71 versus 6.59±1.37, P<0.05), which was similar to that of nature valve tissue. The ultimate tensile stress of glutaraldehyde-fixed group was higher than that of 2,3-butanediol post-treatment group (P<0.005). Histological examination revealed some changes in structural organization of the collagen, which explained the alterations in the biomechanical properties. Conclusion The 2,3-butanediol post-treatment is a better method of treating bovine pericardial, which improved the biomechanical properties of bovine pericardial.