The Maillard reaction and its fluorescent products have attracted widespread attention in the field of food safety and biology. Herein, the novel Maillard reaction fluorescent nanoparticles (MRFNs) as a fluorescent probe were synthesized via a “green” method with simple technical processes. In addition, the effects of tetracycline (TC) and chlorotetracycline (CTC) representing certain properties of tetracyclines (TCs) on the fluorescence behaviour of MRFNs were studied, respectively. The present study showed that the fluorescence intensity of MRFNs greatly enhanced with a linear increase in the CTC concentration. However, with the gradual increase in the TC concentration, the intensity of MRFNs tended to significantly decrease linearly. Based on this, novel fluorescence analysis methods for the simple and rapid detection of TC and CTC in water bodies were established, respectively. Significantly, the proposed detection methods were successfully adopted for detecting TC and CTC in some environmental water samples. Besides, the possible mechanisms for TC-induced fluorescence quenching and CTC-induced fluorescence enhancement of MRFNs were also discussed, respectively.