An ultrasensitive and selective voltammetric sensor with ultra-trace level detection limit is introduced for idarubicin (IDA) determination in real samples. The as-synthesized nanocomposite was characterized by several techniques, including Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), Raman spectroscopy, Energy-dispersive X-ray spectroscopy (EDX), and Field emission scanning electron microscopy (FESEM). The electrocatalytic performance of the developed electrode was observed by cyclic voltammetry (CV), differential pulse voltammetry (DPV), electrochemical impedance spectroscopy (EIS), and chronoamperometry. The limit of detection (LOD) of the developed sensor for idarubicin is 1.0 nM, and the response is found to be in the dynamic concentration range of 0.01-1.9 mu mol/L in a Britton-Robinson buffer (B-R, pH = 6.0). Moreover, the fabricated electrode illustrated high selectivity with good repeatability and reproducibility for diagnosing idarubicin as an anthracycline antileukemic drug. Furthermore, to evaluate the validity of the recommended method, three real samples, including human plasma, urine, and water samples, were analyzed with satisfactory recovery and compared with high-performance liquid chromatography (HPLC). The minor groove-binding mode of interaction was also supported by docking simulation studies, emphasizing that IDA can bind to ds-DNA preferably and confirmed experimental results. The reduced assay time and the possibility of measuring a single sample with another anticancer drug without any interference are significant advantages compared to the HPLC. The developed and validated sensor could be a valuable point-of-care diagnostic tool for IDA quantification in patients.