Fatigue failure is one of the most common damage types of monopiles, which are widely used in offshore wind turbines (OWT). In this paper, a systematic study is performed to investigate the influence of stiffness matrix, sea states and wave directions on fatigue loads of monopiles. The stiffness matrix of the monopile determines natural frequency of the whole system and has great effect on fatigue loads. The influence of the pile diameter and length on the stiffness matrix are analyzed, which are calculated based on p-y curve of foundation. Then, three methods for selected sea states are compared. The dynamic response of the monopile using the different selection method under same wind condition are calculated and their contribution to the fatigue load is discussed. Finally, bending moment on the top of monopiles in vary wave direction are analyzed which shows significant on the fatigue. Several suggestions for design of monopile are given to avoid fatigue failure based on this study.