To determine allergen beta-lactoglobulin (β-LG) in foods, a method by a coffee-ring effect (CRE)-based paper sensor chip combined with a smartphone has been developed. The strategy was based on the principle of fluorescence quenching by the quantum dot-aptamer-graphene oxide (QD-Apt-GO) and where the degree of quenching was inversely proportional to the concentration of β-LG. First, the prepared β-LG solution was added to the QD-Apt fluorescent probe solution followed by an appropriate amount of GO. After the reaction, a certain amount of the mixture was dispensed onto the detection zone of the sensor chip, resulting in the spread of the analyte solution by evaporation-driven flow. The QD-Apt and the QD-Apt-β-LG in the mixed solution formed a visible fluorescent coffee-ring. For signal quantitation, an image of the coffee-ring was taken by a smartphone and imported into self-programmed software for analysis. The quantitative results for β-LG were acquired rapidly based on the magnitude of the generated fluorescent coffee-ring. The limit of detection (LOD) for β-LG was 0.048 mg L-1 and the linear range for the optimized assay was 0.39–1000 mg L-1. The method was validated for analysis of real food samples and the results were consistent with those of a commercial ELISA assay kit. The smartphone-based assay provides a simple, economic, compact, and visual detection platform for the determination of β-LG in foods.