Iron oxide-Copper oxide nanoparticles composite (Fe2O3CuO NPs) was synthesized through a green phytosynthetic approach using Ocimum sanctum Linn (commonly known as Tulsi) leaf extract. The evaluation of electrocatalytic properties were evaluated by carrying out electrochemical detection of amiprofos-methyl (APM), an organophosphorus herbicide. It is moderately toxic to mammals and aquatic biodiversity and is considered to be an acetylcholinesterase inhibitor. The presence of specific natural phytochemicals such as eugenol, naringenin, apigenin, quercetin, and high amount of ascorbic acid in the aqueous extract of Ocimum sanctum Linn plant parts, has been widely used for the synthesis of various metallic nanoparticles where these compounds serve as reducing, stabilizing, and capping agents. The synthesized Fe2O3CuO NPs were characterized using scanning electron microscope (SEM), energy dispersive X-ray analysis (EDX), X-ray diffraction analysis (XRD), UV–Vis spectroscopy, Fourier transform infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA). The modified electrode was electrochemically characterized by cyclic voltammetry and differential pulse voltammetry (DPV) techniques for the detection of APM. The electrochemical signals have increased by three folds in the detection of APM with Fe2O3CuO nanocomposite compared to the bare glassy carbon electrode. The electrochemical sensor showed a linear range of 0.05 to 30 µg/mL with a limit of detection of 0.0065 µg/mL. The developed electrochemical sensor was successfully applied for the detection of APM in different water and soil samples with recoveries ranging from 96.00−99.00%. The electrode showed good stability and reproducibility over a period of 10 days with a 95% of peak current than the former. The newly synthesized nanoparticles, thus, proved to be an interesting material for electrochemical and biological studies.