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In the present study, we utilized a novel electrochemical sensor for the determination of colchicine in 0.1 M phosphate buffer solution (pH 4.0) using differential pulse voltammetry. 1-butyl-3-methylimidazolium tetrachloroferrate ([Bmim]FeCl4) was synthesized, investigated by FT-IR, Raman and UV–Vis spectrometer and applied as magnetic ionic liquid in electrode modification. The other modifier was copper oxide nanoparticles/carbon nanofiber composite (CuO/CNF) provided by electrospinning and evaluated by scanning electron microscopy, X-ray diffraction and Fourier transform infrared spectroscopy (FT-IR). Several electro-analytical parameters such as the electrochemical behavior of the modified electrode, scan rate, type and supporting electrolyte pH were studied for the determination of colchicine. Under the optimized experimental conditions, oxidative peak current increased linearly by increasing colchicine concentration in the range of 1.0 to 100.0 nM and detection limit was obtained 0.25 nM. The relative standard deviation for six replicate analyses of 20.0 nM and 58.0 nM colchicine solution was obtained to be ± 3.17% and ± 2.08%, respectively. The proposed electrochemical sensor was successfully applied for colchicine quantification in colchicine tablet and health human plasma with satisfying results. [ABSTRACT FROM AUTHOR]