In musculoskeletal simulation, individualized joint axes enhance the accuracy and reliability of kinematic and kinetic simulation results. We investigated the correctness and performance of an analytical method for identifying the instantaneous axis of rotation between two bodies based on motion data in OpenSim. The instantaneous center of rotation is the point at which two bodies have the same velocity. The relative linear and angular velocity between the two bodies, as well as their relative position to each another, are required as inputs to calculate it. Using the instantaneous center of rotation, fixed or moving joint centers of rotation can be identified. To prove the general applicability of the method, the instantaneous centers of rotation of a revolute joint of a simple double pendulum model and the hip and knee joint of a more complex musculoskeletal model were investigated. The hip joint is defined as a ball joint. The knee joint is defined as an OpenSim custom joint which describes the motion of the child segment in relation to the parent segment as a function of generalized coordinates. To verify the correctness of the approach in OpenSim, the moving centers of rotation were calculated using synthetic noisefree data. The results were compared to the implementation of the respective joints in the model which act as the ground truth. White Gaussian noise was added to the synthetic data to analyze its effect on the quality of the calculated centers of rotation. We were able to correctly identify the center of rotation of each joint using noisefree data. In the case of noisy data, joint centers of rotation can be determined by applying additional filtering or optimization methods to the calculated instantaneous centers of rotation. Consequently, we are able to determine the center of rotation for arbitrary joints based on noisy synthetic data. This approach is applicable for both fixed and moving centers of rotation which distinguishes it from commonly used methods in the field of biomechanical simulation.