Carbon dots (CDs) were synthesized by refluxing glucose, as efficient fluorescence probes, which show convenient and sensitive detection of norfloxacin (NOR) over a wide concentration range. It is worth noting that because of the hydrogen bond interactions between the CDs and NOR, the fluorescence intensity of CDs was remarkably enhanced in the presence of NOR, which indicates that CDs are capable of rapid, stable and sensitive determination of NOR. Compared with high-performance liquid chromatography, the fluorescence enhancement method is considerably simpler and faster, and will pave a new way for the determination of NOR.