N-doped carbon dots (N-CDs) were synthesized from L-glutamine and triethanolamine via one-step hydrothermal method. The N-CDs emitting blue fluorescence have selective response to tetracyclines (TCs), and can be used as a fluorescent probe to realize the quantitative detection and qualitative analysis of TCs. A method for the determination of TCs by the N-CDs in actual samples has been successfully established. The recovery rate was maintained at 97.50-105.60%, and the relative standard deviation (RSD) was below 3%. Meanwhile, TCs can be visually distinguished using filter paper by the different fluorescence colors (light green, dark blue and yellow-green) of the N-CDs/TCs system under ultraviolet (UV) light. This study provides a relatively simple method to detect and identify TCs.