The consumption of food has an impact on the environment; hence, it is crucial to explore the level of knowledge the urban population possesses regarding the origin of the foods they consume. Objective: To design, validate, and determine the reliability of an instrument to assess the level of knowledge among the urban population regarding agroecological foods. Materials and methods: The study was conducted at the Central University of Ecuador in Quito, Ecuador, during the period 2020-2021. The research type was exploratory-descriptive, with a non-experimental design and stratified non-probabilistic sampling. The instrument’s content was evaluated using the Delphi method with the participation of seventeen experts. Two pilot tests were administered to 370 and 1323 participants, respectively. Internal consistency and homogeneity index were determined using Cronbach’s alpha. The construct’s dimensionality was established using the Cronbach-Mesbah curve. Construct validity was assessed through a factorial analysis. Results: Content validity revealed a convergence value of over 75% among experts. In the validation of internal consistency, the Cronbach’s alpha coefficient was 0.9, and the homogeneity index was greater than 0.2. In the study of construct dimensionality, the Cronbach-Mesbah curve indicated that unidimensionality was fulfilled throughout the instrument. In construct validity through factorial analysis, the Kaiser-Meyer-Olkin measure was 0.753. Bartlett’s sphericity was 0.000, and the factor extraction test was greater than 77%. Conclusions: The instrument demonstrated high validity and reliability, both in its content evaluated by experts and in its internal consistency, dimensionality, and construct validity assessed through statistical methods. The instrument is reliable and can be applied to the urban population.