A fast and facile electrochemical sensor for the detection of an important anticancer drug, 5-fluorouracil, is fabricated using CuFe2O4nanoparticles modified screen printed graphite electrode (CuFe2O4NPs/SPGE). The electrochemical activity of the modified electrode was characterized by chronoamperometry, cyclic voltammetry (CV) and differential pulse voltammetry (DPV) and linear sweep voltammetry (LSV) experiments. The CuFe2O4NPs improved the electrochemical properties of the electrodes and enhanced their electroanalytical performance. Electrochemical measurements using differential pulse voltammetry showed a wide linear relationship between 5-fluorouracil concentration and peak height within the range 0.1 to 270.0 μM with a low detection limit (0.03 μM). Further, the sensor was testified with a urine sample and 5-fluorouracil injection sample, and the observed remarkable recovery results replicate its practical applicability.