Anterior cruciate ligament injury rates are reported to be two to eight times higher in women than in men within the same sport. Because the menstrual cycle with its monthly hormonal fluctuations is one of the most basic differences between men and women, we investigated the association between the distribution of confirmed anterior cruciate ligament tears and menstrual cycle phase. Sixty-nine female athletes who sustained an acute anterior cruciate ligament injury were studied within 24 hours of injury at four centers. The mechanism of injury, menstrual cycle details, use of oral contraceptives, and history of previous injury were recorded. Urine samples were collected to validate menstrual cycle phase by measurement of estrogen, progesterone, and luteinizing hormone metabolites and creatinine levels at the time of the anterior cruciate ligament tear. Results from the hormone assays indicate that the women had a significantly greater than expected percentage of anterior cruciate ligament injuries during midcycle (ovulatory phase) and a less than expected percentage of those injuries during the luteal phase of the menstrual cycle. Oral contraceptive use diminished the significant association between anterior cruciate ligament tear distribution and the ovulatory phase.