Porphin-based carbon dots (denoted as PCDs) are prepared through a one-step hydrothermal method by using meso-tetra (4-carboxyphenyl) porphin (TCPP), citric acid, and ethanediamine as precursor. PCDs give rise to the optimal photoluminescence at λex/λem = 375/645 nm, exhibit an excitation-independent property, excellent water solubility, and good biocompatibility, which provide red emission and avoid the autofluorescence as an efficient fluorescent imaging probe. On the other hand, when Eu3+ is added into PCDs, the carboxylate groups located on the surface of PCDs exhibit high affinity to Eu3+, resulting in the fluorescence of PCDs turning off via static quenching. In the presence of phosphate, owing to the strong coordination with Eu3+, the fluorescence of PCDs turns on. Based on this performance, a novel “turn off–on” phosphate sensing system is developed. The detection limit of this sensing system can attain 3.59 × 10−3 μmol L−1. This system has been utilized for the detection of phosphate in real samples successfully, which further demonstrates potential applications in biological diagnostic and environmental analysis.