Instruments for the measurement of human sexuality include self-report measures used to assess sexual functioning, but many of them have not yet been validated. The Center of Applied Psychology Female Sexual Questionnaire (CAPFS-Q) is an original self-report instrument. It has been developed for the study of sexuality in specific non-clinical populations, such as female university students of Medicine and other Health Sciences. The CAPFS-Q includes 26 items, organized as follows: sociodemographic and relevant data (four items); aspects of sexual relations with partner (five items); sexual practices (12 from 13 items); and dysfunctional aspects of sexual relations (four items). CAPFS-Q validity and reliability were examined in a sample of Spanish female university students of Health Sciences. Exploratory and confirmatory factor analysis (FA) showed a four-factor structure which explained 71.6% of the variance. This initial version of the CAPFS-Q is a reliable measure of women’s sexual behavior, with a dimensionality that replicates the initial theoretical content and with adequate indicators of internal consistency, validity, and test–retest reliability. It is easy to administer and to complete.