Hamstring strains are the most common time-loss injury in elite Gaelic football affecting over 20% of players per season. Thus, there is a need to identify factors contributing to the onset of hamstring injuries in order to inform injury risk management strategies. The current study investigated whether eccentric knee flexor strength and between-limb imbalances were associated with increased risk of sustaining a time-loss hamstring injury in elite Gaelic football players. A total of 185 elite male players (26.9 ± 2.7 years, 86.4 ± 6.2 kg, 183.4 ± 5.6) were prospectively followed for 12 weeks from the day of testing. Injury data were provided by the team medical staff. Twenty-eight players (16%) sustained a time-loss hamstring injury following testing. Players that did not sustain a hamstring injury had greater average between-limb asymmetries (uninjured = 9.1%, 95% CI 7.8-10.1; injured = 5.1%, 95% CI 3.7-6.7; P = .001). Eccentric knee flexor strength profiles were not associated with increased or decreased risk of sustaining a hamstring injury and did not alter the post-test probability of sustaining a hamstring injury across the investigation period. These findings do not support the use of eccentric knee flexor strength metrics in managing hamstring injury risk in elite male Gaelic football players.