Background. Non-contact anterior cruciate ligament (ACL) injuries occurs with a higher incidence in female athletes compared with males after the onset of puberty. One anatomical factor with clinically observable differences between males and females is lower extremity alignment. The knee joint valgus in the coronal plane, which is associated with ACL injury risk, is composed of rotation of the tibia around the stationary femur and hip rotation combined with knee flexion. The purpose of this study was to prove the difference of hip rotation between female and male handball players with or without history of ACL injury. Methods. Elite collegiate athletes on the varsity handball team (17 females and 24 males) were recruited. Whereas 8 females and 1 male had a history of ACL injury, there were 9 females and 23 males with no history of ACL injury. The 6 measures of hip joint motion [flexion, extension, abduction, adduction, external rotation (ER), and internal rotation (IR)] and the 2 measures of knee joint motion (flexion and extension) were measured with the standard procedure using a goniometer. Results. The range of ER in ACL-injured females (37±7.5) was smaller than in uninjured females (49±8.2). The range of IR in ACL-injured female players (55±9.6) was greater than in uninjured females (41±13). There was no difference in ER+IR between ACL-injured and uninjured females. In the ACL-injured male handball player, ER was dominant to IR. Compared with the uninjured males, the ACL-injured female players showed smaller range of ER, greater ranges of IR and greater ER+IR. Conclusions. ACL-injured female handball players had greater hip IR with smaller hip ER, compared with uninjured counterpart. Hip rotation may be a crucial risk factor for ACL injury in female athletes. Level of evidence. 2b