Background: For gendered innovations to be streamlined in health research, researchers need tools to measure gender variables quantitatively. Because gender is a sociocultural concept which varies by culture, it is important to check feasibility of using tools developed in western countries before using them in Korea. The objective of this study is to conduct a pilot study for adapting a questionnaire measuring gender variables from the literature for the Korean population. Methods: The research was conducted in two steps: 1) modification of questionnaire by an expert group, and 2) an on-line survey of 3,000 adults, in three age groups, balanced in sex. The questionnaire was composed of 7 gender variables with 2 to 6 items, constituting a total of 24 5-point Likert scale items. Gender variable scores were calculated as the average of Z-scores of items pertaining to each variable. Results: Participants were 45.6% men, 43.9% women, and 10.5% other gender. Scores of five gender variables-caregiver tension, work tension, independence, emotional intelligence, and discrimination-were significantly different among sex and gender groups. Among gender variables, work strain, caregiver strain, and discrimination had notable influences on health status and health-related behaviors among survey participants. Conclusions Several gender variables were related to health status and health behaviors in Korean adults. This is the first attempt in Korea to quantify gender characteristics to be used for health research. Further studies are needed to verify the questionnaire and methods developed in this study for use with diagnosed disease outcomes.