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In the present work, the simultaneous determination of dexamethasone (DXM) and hydrocortisone (HYD) is described by the use of a nano-porous glassy carbon electrode (GCE), prepared via an electrochemical treatment. Nano-porous/GCE provided the porosity which was increased the sensitivity of the surface of electrode to monitor the drugs. Differential pulse voltammetry (DPV) was used for the determination of DXM and HYD simultaneously at the surface of the electrode. Under the optimum pH 2 (0.1 M, phosphate buffer solution), it is found that the calibration plots for DXM and HYD are linear in the concentration ranges 0.02–22 and 0.1–42 µM with detection limits of 5 and 30  nM for DXM and HYD, respectively. Also, the interaction of human growth hormone (somatropin) with DXM and HYD has been studied by electrochemical and UV–Vis spectroscopy techniques. The binding of DXM and HYD to human growth hormone was also modeled by molecular docking methods. The glucocorticoids could inhibit the application of the human growth hormone.