AgCu nanoparticles were prepared through hydrogen-reduction-assisted Ultrasonic Spray Pyrolysis (USP) and the Hydrogen Reduction (HR) method. The changes in the morphology and crystal structure of nanoparticles were studied using different concentrated precursors. The structure and morphology of the mixed crystalline particles were characterized through X-ray diffraction analysis (XRD), scanning electron microscopy (FEG-SEM), transmission electron microscopy (TEM) and Energy-dispersive X-ray spectroscopy (EDS). The average particle size decreased from 364 nm to 224 nm by reducing the initial solution concentration from 0.05 M to 0.4 M. These results indicate that the increase in concentration also increases the grain size. Antibacterial properties of nanoparticles against Escherichia coli were investigated. The obtained results indicate that produced particles show antibacterial activity (100%). The AgCu nanoparticles have the usage potential in different areas of the industry.