Purpose: The aim of this study was to develop a reliable measurement tool to assess the urban food environment, particularly focusing on food stores in Korea. Methods: The items for the measurement tool were selected through systematic literature reviews and adjusted to the Korean food environment. A total of 25 food stores in an urban area were recruited for the pilot test to evaluate the time required for completion of the survey, ease of response, willingness to participate, difficulties in conducting the survey, and appropriateness, and 34 food stores were recruited for assessment of the reliability of the tool using percent agreement and kappa value. Results: The measurement tool is composed of questions regarding food store characteristics, accessibility, and food availability. On average, 26 minutes was required to survey a single food store, and the subjects and interviewers answered that the process was not difficult for the survey. The percent agreement for the inter-rater and test-retest reliability ranged from 93.9~98.8% and 91.9~97.9, respectively. The kappa values ranged from 0.78 to 0.97, which was very high. Percent agreement and kappa value of food with healthy option were lower than those of the general food in the inter-rater reliability (p = 0.0027, p = 0.0095 respectively) as well as in the test-retest reliability (p = 0.0081, p = 0.0290, respectively), although they were still high enough (86.4~98.0% for percent agreement, 0.64~0.96 for kappa value). Conclusion: The newly developed measurement tool for assessment of food store environment appears to be feasible and reliable; therefore, it can be applied to research on the association between food environment and dietary behaviors as well as health outcomes.