In the current study, DNA immobilization was performed on pencil graphite (PG) modified with a polypyrrole (PPy) and flower-like Pt/NiCo2O4 (FL-Pt/NiCo2O4) nanocomposite, as a new sensitive electrode to detect chlorambucil (CHB). Energy dispersive X-ray (EDX) analysis, X-ray diffraction (XRD) and scanning electron microscopy (SEM) techniques were employed to characterize the synthesized FL-Pt/NiCo2O4 and PPy/FL-Pt/NiCo2O4 nanocomposites. Moreover, differential pulse voltammetry (DPV) was selected to assess the guanine and adenine electrochemical responses on the DNA sensor. The CHB determination was performed using the maximum currents towards adenine and guanine in the acetate buffer solution (ABS). According to the results, ds-DNA/PPy/FL-Pt/NiCo2O4/PGE was able to detect the different concentrations of CHB in the range between 0.018 and 200 μM, with a detection limit of (LOD) of 4.0 nM. The new biosensor was also exploited for CHB determination in real samples (serum, urine and drug), the results of which revealed excellent recoveries (97.5% to 103.8%). Furthermore, the interaction between ds-DNA and CHB was studied using electrochemistry, spectrophotometry and docking whose outputs confirmed their effective interaction.
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