In this study, biological synthesis of silver nanoparticles (AgNPs) using Senna auriculata flower extract for antibacterial activities was reported. The silver spectra compared to the plant extract show a rightward shift in AgNP peaks, indicating successful nanoparticle formation. The absorption band at 302 nm and the disappearance or shift of other peaks further confirm the synthesis. X‐ray diffraction (XRD) analysis reveals that the AgNPs synthesized with S. auriculata extract have an average crystallite size of 25 nm. Transmission electron microscopy (TEM) results exhibited a polydispersed, spherical shape with sizes ranging from 70 nm, in clear contrast to the electron microscope image that showed their spherical shape. When examining the selected area electron diffraction (SAED) image, a specific set of lattice planes was correlated with a specific spot. A histogram of AgNP particle size distribution can be seen. AgNPs were tested against four different strains of bacteria for their antibacterial effectiveness, including gram negative bacteria (Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus), as well as gram positive bacteria (S. aureus, d. Bacillus subtilis), at various concentrations of AgNP. The results of in vitro experiments indicate that AgNPs containing S. auriculata flowers inhibit amylase well. At two concentrations, ~16.03% and ~70.99%, AgNPs inhibit the reaction at low and high concentrations, respectively.