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In this study, a screen-printed electrode (SPE) modified with gold-nanoparticle-decorated reduced graphene oxide–carbon nanotubes (rGO-AuNPs/CNT/SPE) was used for the determination of estradiol (E2). The AuNPs were produced through an eco-friendly method utilising plant extract, eliminating the need for severe chemicals, and remove the requirements of sophisticated fabrication methods and tedious procedures. In addition, rGO-AuNP serves as a dispersant for the CNT to improve the dispersion stability of CNTs. The composite material, rGO-AuNPs/CNT, underwent characterisation through scanning electron microscopy (SEM), ultraviolet–visible absorption spectroscopy (UV–vis), Fourier-transform infrared (FTIR) spectroscopy, and atomic force microscopy (AFM). The electrochemical performance of the modified SPE for estradiol oxidation was characterised using cyclic voltammetry (CV) and differential pulse voltammetry (DPV) techniques. The rGO-AuNPs/CNT/SPE exhibited a notable improvement compared to bare/SPE and GO-CNT/SPE, as evidenced by the relative peak currents. Additionally, we employed a baseline correction algorithm to accurately adjust the sensor response while eliminating extraneous background components that are typically present in voltammetric experiments. The optimised estradiol sensor offers linear sensitivity from 0.05–1.00 µM, with a detection limit of 3 nM based on three times the standard deviation (3δ). Notably, this sensing approach yields stable, repeatable, and reproducible outcomes. Assessment of drinking water samples indicated an average recovery rate of 97.5% for samples enriched with E2 at concentrations as low as 0.5 µM%, accompanied by only a modest coefficient of variation (%CV) value of 2.7%.

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