Aiming at the problem of the influence of preloading force on its mechanical response in soft tissue compression experiments, an elimination method of preloading force based on linear loading region is proposed. Unconfined compression experiments under a variety of different preloading forces are performed. The influence of the preloading force on the parameters of constitutive model is analyzed. In the preload phase, the mechanical response of the soft tissue is taken as a linear model. The preloading force is eliminated by taking the preloading phase into account throughout the response process. According to five different preloading forces of the unconfined compression experiments, the elimination method is validated with two different constitutive models of soft tissue, and the error between the models obtained by the preloading force elimination method and the traditional method with the experimental results is compared. The results show that the error obtained by preloading force elimination method is significantly smaller than the traditional method. The preloading force elimination method can eliminate the influence of preloading force on mechanical response to a certain extent, and constitutive model parameters which are closer to the true properties of soft tissue can be obtained.