Handball is a strenuous body-contact team sport that places high loads on the knee joint. Anterior cruciate ligament (ACL) tear is one of the most devastating injuries that any handball player can suffer, and female athletes are at particular risk due to their intrinsic anatomical, hormonal, neuromuscular and biomechanical characteristics. The purpose of this study was to analyze the horizontal jumping biomechanics of female elite handball players with or without previous ACL reconstruction.Twenty-one female participants (6 with previous ACL reconstruction and 15 uninjured controls) were recruited. Two horizontal hopping tasks were evaluated using inertial sensor unit (ISU)-based technology to assess jumping biomechanics through a direct mechanics-based approach.The athletes with previous ACL reconstruction demonstrated a significant (P < 0.05) reduction in the unilateral triple hop for distance compared with the healthy controls. Furthermore, during the initial propulsive phase of the unilateral cross-over hop, the control participants generated significantly (P < 0.05) higher force values in the mediolateral direction (the X axis) with their dominant limb compared with the ACL-reconstructed (ACL-R) limb of previously injured participants.Three-dimensional horizontal jumping biomechanics analyses using ISU-based technologies could provide clinicians with more accurate information regarding the horizontal jumping biomechanical patterns among elite handball female athletes. Furthermore, several mechanical alterations could still be observed among those players who had undergone previous ACL reconstruction, even when several years have passed since the original ACL injury.