The assessment of eating and physical activity habits is an important step in promoting healthy behaviors among the adolescent population and is key in the prevention and management of chronic non-communicable diseases, such as obesity, diabetes, and cardiovascular disease. For this purpose, reliable and valid measuring instruments are essential. In this context, the aim of this article is to present the validation of a self-report questionnaire on eating and physical activity habits among adolescents in Mexico City. In order to validate the questionnaire, a cross-sectional study was conducted with a sample of 2710 adolescents between 11 and 12 years of age, the piloting of the questionnaire was carried out in September 2022 with a focus group, and the programming of the anthropometric measurements was established with the Federal Educational Authority of CDMX, as well as the application of the questionnaire to 33 schools, with these activities being scheduled from 7 November 2022 to 3 February 2023 and having an application duration of 15–25 min for each of the groups to which it was applied; the questionnaire that was applied consists of 31 questions that refer to the frequencies, quantity, or performance of behaviors related to the frequency and type of food, type of physical activity and behaviors related to the act of eating referring to the place where it is carried out (home or away from home) and with whom it is carried out (alone or in company), and about the individual’s lifestyle. Subsequently, the reliability of the instrument was evaluated using Cronbach’s alpha coefficient, and an exploratory factor analysis was conducted to determine the structure of the questionnaire. The results obtained showed that the questionnaire was adequately reliable (α = 0.778) with an eight-factor structure: four questions on mealtime frequencies, four questions on physical activity and lifestyles, six questions on the consumption of high-calorie foods, four questions on company and food consumption, four questions on the consumption of vegetables and fruits, four questions on the place of food consumption, two questions on the consumption of alcoholic beverages, and three questions on the consumption of sugary drinks, plain water, and milk. In conclusion, the self-report questionnaire on eating and physical activity habits among adolescents in Mexico City is reliable, has adequate internal consistency, and can therefore be used as a useful tool for the evaluation of eating and physical activity habits in this population.