Recent research has revealed that lifestyle behaviors and dietary habits put athletes at equal risk as the general population of hypovitaminosis D. A blood test, the gold standard for measuring vitamin D, is costly and invasive. A non-invasive, reliable, and valid measure that estimates the vitamin D intake of female college athletes, provides a tool for assessing the risk of hypovitaminosis D in this population of athletes. The purpose of this study was to assess the validity and reliability of a self-reported Vitamin D-Specific Food Frequency and Lifestyle Questionnaire (VDFFLQ) developed to evaluate the vitamin D intake of female college athletes.