Green and low-cost synthesis strategy for ultrasonic preparation of polymer blend matrix based silver nanoparticles (Ag NPs) and the development of rapid and high sensitive detection route have a great attention in biomedical applications. Therefore, in this study, we investigated the hydrogen peroxide detection performance of Konjac gum (KG)/PEG-Ag NPs. The KG/PEG-Ag NPs was synthesized via an ultrasonic process and characterized by different techniques such as ultraviolet–visible spectroscopy (UV–Vis), Fourier-Transform Infrared spectroscopy (FT-IR), Scanning Electron Microscope (SEM) and Energy Dispersive X-ray spectroscopy (EDX). Furthermore, we determined the experimental optimization on the effect of the rheological parameters of nanostructure with the highest correlation constant (R 2 : 0.989-0.996), and the intrinsic viscosity (14.71-26.77 dl/g). To provide the miscible polymer blends and homogeneous dispersion of the nanostructure, we compared the rheological parameters with the experimental results. The response time was less than 5 s and the lower limit of detection was 0.071 μM. This novel highly sensitive, rapid, and naked-eye colorimetric biosensor based Ag NPs which are prepared ultrasonic manufacturing approach, opens up a green approach of development facile and rapid detection of hydrogen peroxide in practical biomedical applications.