A questionnaire on explanatory variables for each behavior of the 24-h movement behaviors (i.e., physical activity, sedentary behavior, sleep) was developed based on three levels of the socio-ecological model, i.e., the intrapersonal level, interpersonal level and the physical environmental level. Within these levels, different constructs were questioned, i.e., autonomous motivation, attitude, facilitators, internal behavioral control, self-efficacy, barriers, subjective norm, social modeling, social support, home environment, neighborhood, and work environment. The questionnaire was tested for test-retest reliability (i.e., intraclass correlation (ICC)) for each item and internal consistency for each construct (i.e., Cronbach's Alpha Coefficient) among a group of 35 healthy adults with a mean age of 42.9 (±16.1) years. The total questionnaire contained 266 items, consisting of 14 items on general information, 70 items on physical activity, 102 items on sedentary behavior, 45 items on sleep and 35 items on the physical environment. Seventy-one percent of the explanatory items showed moderate to excellent reliability (ICC between 0.50 and 0.90) and a majority of constructs had a good homogeneity among items (Cronbach's Alpha Coefficient ≥ 0.70). This newly developed and comprehensive questionnaire might be used as a tool to understand adults' 24-h movement behaviors.