Monitoring of herbicides and pesticides in water, food, and the environment is essential for human health, and this requires low-cost, portable devices for widespread deployment of this technology. For the first time, a wearable glove-based electrochemical sensor based on conductive Ag nano-ink was developed for the on-site monitoring of trifluralin residue on the surface of various substrates. Three electrode system with optimal thicknesses was designed directly on the finger surface of a rubber glove. Then, fabricated electrochemical sensor used for the direct detection of trifluralin in the range of 0.01 μM to 1 mM on the surface of tomato and mulberry leaves using square wave voltammetry (SWV) and difference pulse voltammetry technique. The obtained LLOQ was 0.01 μM, which indicates the suitable sensitivity of this sensor. On the other hand, this sensor is portable, easy to use, and has a high environmental capability that can be effective in detecting other chemical threats in the soil and water environment.