In this work, sulfur and nitrogen co-doped carbon quantum dots (S,N-CQDs) were prepared via one-pot hydrothermal treatment of EDTA disodium and sodium sulfide. The prepared S,N-CQDs were characterized by TEM, XRD, FT-IR, XPS, UV-vis absorption and fluorescence spectra to characterize their morphology, crystal structure, functional groups, elemental composition, and optical properties. It was found that S and N elements were successfully doped into the CQDs and the morphology was approximately spherical with an average particle size of 2.16 nm, in which the excitation/emission wavelengths were 350 and 420 nm, respectively. Compared with single element doped CQDs, double element doped CQDs have a higher quantum yield and excellent optical stability. Cell experiments showed that S,N-CQDs had good biocompatibility because they had no obvious toxicity on both normal cell lines and cancer cell lines. More importantly, based on the synergy of static quenching and dynamic quenching, the S,N-CQDs were used as effective fluorescent probes for sensitive detection of DA, with high anti-interference and low limit of detection. Based on the good biocompatibility of S,N-CQDs, the detection of dopamine in actual serum samples were carried out and the results showed an excellent recovery rate. Therefore, this work provides a dopamine sensor with a practical application prospect.