Precise monitoring of pesticide with portable device was challenging because it required high sensitivity, short response time, strong stability and excellent selectivity. Herein, we newly constructed a stimuli-responsive hydrogel (SRHg)-based portable kit by embedding copper nanoparticles (CuNPs) in agarose hydrogel. In this work, dimethoate as inhibitor of urease restrained the generation of ammonia, which reduced in-situ etching of CuNPs, resulting in the fluorescence color response of test kit under ultraviolet illumination. Interestingly, by means of smartphone-based nanocolorimetry, the photo image of portable kit could be translated into digital information using ImageJ software, achieving a direct quantitative tool for dimethoate identification. The simplicity of SRHg-based portable kit combined with smartphone-based color recognition not only improved the analysis sensitivity (detection limit of 1.0 μg L−1 ), accuracy and stability, but also simplified operation process and shortened sample-to-answer analysis time (55 min), demonstrating that the methodology met the needs of daily testing and provided a new sight for on-site monitoring of food safety and human health.