Sudan III, a synthetic azo dye, is commonly used for coloring oils, waxes, and various plastics. However, its unauthorized presence in food and food products poses serious health hazards, including potential carcinogenic effects. In this study, we investigated the application of nitrogen and sulfur co-doped carbon quantum dots (N,S-CQDs) as a fluorescence sensor to detect trace amounts of sudan III. When exposed to trace amounts of sudan III, the fluorescence signal of N,S-CQDs was significantly enhanced. Under optimal conditions (pH 5.0 and an incubation time of 4 minutes), the fluorescence enhancement of N,S-CQDs showed a linear relationship with sudan III concentration in the range of 7.5 x 10-10 to 1.0 x 10-8 M, with a limit of detection of 2.0 x 10-10 M (equivalent to 0.07 ppb). The method demonstrated high accuracy (recovery from 91.5% to 103.5%), good reproducibility (RSD < 3.86%), and excellent selectivity for sudan III over other azo dyes. We successfully applied this developed method to analyze sudan III in chili sauce and chili powder samples, cross-checking differences between results less than 10% with LC-MS/MS analysis.