The aim of this study was to investigate the effect of a multi-directional soccer-specific fatigue protocol on sprinting kinematics in relation to hamstring injury risk. Nine semi-professional soccer players (Mean±SD: Age: 21.3±2.9 year; Height 185.0±8.7 cm; Body Mass 81.6±6.7 kg) completed the SAFT90; a multi-directional, intermittent 90 min exercise protocol representative of soccer match-play. The 10m sprint times and three-dimensional kinematic data were recorded using a high-speed motion capture system (Qualisys Track Manager®) every 15 min during the SAFT90. A significant time dependent increase was observed in sprint time during the SAFT90 (P<0.01) with a corresponding significant decrease in stride length (P<0.01). Analysis of the kinematic sprint data revealed significantly reduced combined maximal hip flexion and knee extension angle, indicating reduced hamstring length, between pre-exercise and half-time (P<0.01) and pre-exercise and full-time (P<0.05). These findings revealed that the SAFT90 produced time dependent impairments in sprinting performance and kinematics of technique which may result from shorter hamstring muscle length. Alterations in sprinting technique may have implications for the increased predisposition to hamstring strain injury during the latter stages of soccer match-play.