To assess the validity of the Market Basket Analysis Tool (MBAT) for food environment quality within various retail environments compared to the Nutrition Environment Measures Survey in Stores (NEMS-S). In-store assessments using the MBAT and the NEMS-S on the same day in a given store were conducted in grocery stores, corner stores, pharmacies, and dollar stores in a metropolis, and urban and rural counties across 4 states: Louisiana, Mississippi, North Carolina, and Virginia. Descriptive statistics, correlations, and ANOVAs were used to assess store location, store type differences, and MBAT and NEMS-S scores. Market Basket Analysis Tool and NEMS-S data were collected from 114 stores. Market Basket Analysis Tool and NEMS-S total and all individual component scores were significantly correlated (r = 0.84, P ≤ 0.0001; r range, 0.51–0.88; P ≤ 0.0001). The MBAT offers a methodology to measure the food retail environment focusing on the availability of healthful food items with a reduced training time and streamlined data collection compared with the NEMS-S. Future work can assess the completion time of the MBAT compared with the NEMS-S and the ability of the MBAT to detect changes in food environment quality post healthy food retail interventions.