Force control abilities are essential to interact with objects in our environments. However, there is a lack of evaluation tools and methods to test the force control abilities of the upper limb in evaluating the upper limb functions of prosthetic users. This study aimed to quantify upper limb isometric force control abilities in healthy individuals and prosthetic users using a custom-built handle with a 6-axis force/torque sensor and visual cue, namely an Upper Limb End-effector type Force control test device (ULEF). Feasibilities of the test device were demonstrated through experiments by holding the ULEF with an intact hand among healthy subjects and transradial and wrist amputees with a myoelectric powered prosthetic hand, the bebionic hand. Compared to the healthy individuals, the prosthetic user group demonstrated poor isometric force control abilities in terms of higher control instability during the lateral direction task ( [Formula: see text]). Significantly higher variability in force-generating rates was also found in all task directions in the prosthetic user group ( [Formula: see text]). Compared to the healthy group, the prosthetic user group showed significant small peak biceps activities during the posterior task ( [Formula: see text]) and anterior task ( [Formula: see text]). Quantification of isometric upper limb force control abilities can potentially be beneficial to develop evaluation and research tools for investigating mechanisms underlying force control abilities of prosthetic users and provide guidelines for targeted isometric force control training and prosthesis development.