Anterior cruciate ligament (ACL) injury is one of the most serious time-loss injuries in football. The purpose of the present study is to evaluate risk factors of ACL injuries in high-level football players during the preseason. Thirty-five high-level players were captured by a video movement analysis system and a two-force platform. Frontal plane knee angle and vertical force were measured during the landing phase of the drop jump. Intraclass correlation coefficient was determined to assess test-retest reliability, and α level of 0.05 was considered statistically significant. There were significant differences between the stronger and weaker leg in eccentric (32.4 Ns; 95% CI; 22.9-41.8 Ns; P < .05) and concentric impulse (14.4 Ns; 95% CI, 9,8-19,1 Ns; P < .05). Additionally, 31.4% of the subjects obtained a lower limb strength imbalance higher than 15%. Furthermore, 14.2% of the subjects obtained a frontal plane knee valgus angle higher than 20°. Findings of the present study are particularly significant as a functional deficit higher than 15% and an increased in valgus motion at the knee suggests an altered muscular control of the lower extremity. Furthermore, results confirmed that the test proposed in this study is reliable and sensitive to identify ACL injury intrinsic risk factors in high level male football players in order to introduce the necessary preventive measures.