The existence of warfarin is beneficial for health, but its increase in the human body causes toxicity and increases the risk of bleeding. This research focuses on introducing a facile and safe fluorescence sensor for warfarin detection in biological samples. The synthesis of the fluorescence sensor probe was easily performed with doping metal ions of the Pt to CdTe quantum dots. The morphological and optical properties of the synthesized Pt:CdTe quantum dots were characterized by FT-IR, TEM, and EDX. The interaction patterns of warfarin with CdTe QDs were investigated by the quantum chemical method and compared with experimental results. The proposed quantum dots exhibited a blue luminescence with a 28.8% quantum yield. Pt:CdTe QDs were used as the fluorescence probe to assay warfarin. This procedure is based on the "off" fluorescence of quantum dots in the presence of different concentrations of warfarin. Under optimal conditions, the fluorescence sensor probe could detect the concentration of warfarin with a wide linear range of 0.1–100 μM and a detection limit (S/N = 3) of 0.05 µM. Results of sample analysis by fluorescent nanoprobe displayed that this probe could be the potential alternative tool for warfarin detection in biological samples.