BACKGROUND: We tested the hypothesis that impulsive compression, flexion and valgus knee moment loading during a simulated one-footed jump landing will significantly increase the peak relative strain in the anteromedial region of the anterior cruciate ligament compared with loading without the valgus moment. METHODS: Ten cadaveric knees [mean (SD) age: 67.9 (7.6) years; 5 males; 5 females] were mounted into a custom fixture to simulate a lower extremity impact loading of approximately 1600 N. Triaxial load cells monitored the 3D tibial and femoral impulsive force and moments at 2000 Hz, while 3D tibiofemoral kinematics were measured at 400 Hz. Pre-impact quadriceps, hamstring and gastrocnemius muscle forces were simulated using pretensioned steel cables. A differential variable reluctance transducer measured the relative strain in the anteromedial aspect of the anterior cruciate ligament. With the knee initially in 25 degrees flexion, 10 trials were conducted with the impulsive force directed 4 cm posterior to the knee joint center in the sagittal plane ("neutral" loading) to cause a flexion moment, 10 trials were conducted under a similar loading, but with the force directed 15 degrees lateral to the knee sagittal plane ("valgus" loading), and the 10 neutral loading trials were then repeated. A non-parametric Wilcoxon signed rank test was used to test the hypothesis using a P<0.05 significance level. FINDINGS: The peak normalized anterior cruciate ligament strain was 30% larger for the impulsive compression loading in valgus and flexion compared with an impulsive compression loading in isolated flexion (P<0.05). INTERPRETATION: Minimizing the abduction loading of the knee during a jump landing should help reduce anterior cruciate ligament strain during that maneuver.