Glyphosate (N-(phosphonomethyl)glycine) is well known nonselective and broad-spectrum herbicide that has been extensively used in agricultural areas around the world to increase agricultural productivity. However, the utilization of glyphosate can cause environmental contamination and health problems. Therefore, the detection of glyphosate with a fast, low-cost, and portable sensor is still important. In this work, the electrochemical sensor has been developed by modifying of working surface on the screen-printed silver electrode (SPAgE) with a mixtures solution between zinc oxide nanoparticles (ZnO-NPs) and poly(diallyldimethylammonium chloride) (PDDA) by the drop-casting process. The ZnO-NPs have been prepared based on a sparking method by using pure zinc wires. The ZnO-NPs/PDDA/SPAgE sensor shows a wide range of glyphosate detection (0μM-5 mM). The limit of detection of ZnO-NPs/PDDA/SPAgE is 2.84μM. The ZnO-NPs/PDDA/SPAgE sensor exhibits high selective towards glyphosate with minimal interference from other commonly used herbicides including paraquat, butachlor-propanil and glufosinate-ammonium. Furthermore, the ZnO-NPs/PDDA/SPAgE sensor demonstrates a good estimation of glyphosate concentration in real samples such as green tea, corn juice and mango juice.