The objective of this study was to estimate the limitations imposed by the kinetics of activation and relaxation on the ability of slow skeletal muscle to produce mechanical work. These estimates were made by the following methods: 1) using the work loop technique and measuring the actual mechanical work (WA) produced by rat soleus muscles (n = 6) at four different frequencies (0.5, 1, 2, and 4 Hz) and seven different amplitudes of length change (1, 2, 3, 4, 5, 6, and 7 mm); 2) determining the force-velocity relationships of the soleus muscles and using this data to quantify the theoretical mechanical work (WT) that could be produced under the work loop conditions described above; and 3) subtracting WA from WT. The difference between WT and WA was interpreted to represent limitations imposed by activation and relaxation. Under certain conditions (high frequency, small strain), factors controlling the kinetics of activation and relaxation reduced the mechanical work of the soleus muscle by approximately 60%. Hence, activation and relaxation collectively represent important factors limiting the production of mechanical work by slow skeletal muscle.