Objective: Nocturia Quality of Life (N-QOL) questionnaire is a self-administered, nocturia-specific, QoL instrument developed to examine QoL impact of nocturia in male patients. This study was designed to evaluate content and face validity of the N-QOL in female patients. Methods: Twenty women (mean age 59 years; range 27-83) diagnosed with nocturia (=2 voids/night) were recruited through U.S. urology clinics. To establish content and face validity, the items and response options in the questionnaire must be considered relevant and comprehensive. The N-QOL questionnaire was evaluated using 2 methodologies. First, 15 patients provided information on their experiences of the impact of nocturia on their QoL and reported the most bothersome consequences of nocturia in a focus group format. Patient responses were thereafter compared with N-QOL items to evaluate how well they reflected the N-QOL concepts being measured. Second, five patients directly evaluated the N-QOL in an interview format using standardized cognitive debriefing methodology. Results: Of the 20 participants (80% Caucasian, 10% African-American, 10% Hispanic), 45% had their condition for more than 5 years (45% had 3 voids/night and 25% =4 voids/night). Seventy percent had nocturia secondary to OAB, and 30% were currently taking prescription medication for the underlying cause of their nocturia. Disrupted sleep was the most bothersome consequence of nocturia, which resulted in sleeping longer into the daytime hours, being too tired to exercise, eating at night, weight gain, difficulty concentrating, and reduced productivity during daytime. This corresponded well with N-QOL concepts. Directly evaluated the N-QOL was found simple, clear, easy to complete, and comprehensive. Conclusion: The N-QOL has face and content validity in female nocturia patients, with sleep disruption causing severe impact on daytime activities, as the most bothersome consequence. The N-QOL items and response options are relevant and comprehensive for assessing the impact of nocturia on QoL of female patients.