Purpose The depth information of muscle motor units is important in clinical applications. The purpose of this paper is to investigate relationship between motor unit action potentials from surface electromyography and motor unit (MU) depth. Methods Firstly, the skin structure was depicted, and the action potential was considered as a constant current source, which was generated by a tripole model. Secondly, the potential generated by the tripole on the surface was given. Finally, the relationships between MU depth and MU action potential (MUAP) parameters on the skin were studied. Results The regression analysis showed a low relative error for the simulated signals. When the position of the detected electrodes was 18 mm, the linear regression analysis yielded 2.47% in the case of arc, and 2.48% in the case of side. The relationships between the peak to peak amplitude of MUAP and the lengths (arc and side) were tested by the nonlinear regression analysis. The sum of the squared errors was less than 0.0202, and the coefficient of determination was more than 0.9939. Conclusions These results show that it is possible to estimate MU depth by action potentials on surface electrodes.