Repeated applications of glyphosate may contaminate the soil and water and threaten their quality both within the environmental system and beyond it through water erosion related processes and leaching. In this study, we focused on the transport of glyphosate and its metabolite aminomethylphosphonic acid (AMPA) related to soil erosion at two slope gradients (10 and 20°), two rates of pesticide with a formulation of glyphosate (Roundup®) application (360 and 720 mg m− 2), and a rain intensity of 1.0 mm min− 1 for 1 h on bare soil in hydraulic flumes. Runoff and erosion rate were significantly different within slope gradients (p < 0.05) while suspended load concentration was relatively constant after 15 min of rainfall. The glyphosate and AMPA concentration in the runoff and suspended load gradually decreased. Significant power and exponent function relationship were observed between rainfall duration and the concentration of glyphosate and AMPA (p < 0.01) in runoff and suspended load, respectively. Meanwhile, glyphosate and AMPA content in the eroded material depended more on the initial rate of application than on the slope gradients. The transport rate of glyphosate by runoff and suspended load was approximately 14% of the applied amount, and the chemicals were mainly transported in the suspended load. The glyphosate and AMPA content in the flume soil at the end of the experiment decreased significantly with depth (p < 0.05), and approximately 72, 2, and 3% of the applied glyphosate (including AMPA) remained in the 0–2, 2–5, and 5–10 cm soil layers, respectively. The risk of contamination in deep soil and the groundwater was thus low, but 5% of the initial application did reach the 2–10 cm soil layer. The risk of contamination of surface water through runoff and sedimentation, however, can be considerable, especially in regions where rain-induced soil erosion is common.