Pesticides are an important tool for maintaining and improving the global population’s standard of living. However, their presence in water resources is concerning due to their potential consequences. Twelve water samples from rivers, dams/reservoirs, and treated drinking water were collected from Mangaung Metropolitan Municipality in South Africa. The collected samples were analysed using high-performance liquid chromatography linked to a QTRAP hybrid triple quadrupole ion trap mass spectrometer. The ecological and human health risks were assessed by risk quotient and human health risk assessment methods, respectively. Herbicides, such as atrazine, metolachlor, simazine and terbuthylazine, were analysed in water sources. The average concentrations of simazine in rivers (1.82 mg/L), dams/reservoirs (0.12 mg/L), and treated drinking water (0.03 mg/L) were remarkable among all four herbicides detected. Simazine, atrazine, and terbuthylazine posed high ecological risks for both acute and chronic toxicity in all water sources. Moreover, simazine is the only contaminant in the river water that poses a medium carcinogenic risk to adult. It can be concluded that the level of herbicide detected in water sources may affect aquatic life and human beings negatively. This study may aid in the development of pesticide pollution management and risk reduction strategies within the municipality.