Concentrations of widely used antibiotics were predicted to be the highest in the Haihe River Catchment across China previously, potentially resulting in high ecotoxicological risks in this region. As a result of growing usage and regulation of antibiotic use in animals, the pattern of use may have altered temporally for different antibiotics. It is important to monitor the occurrence of antibiotics within different categories for understanding their mass loading to the catchment and the potential ecotoxicological risks involved. This study investigated the seasonal occurrence and spatial variation of 15 antibiotics in the Haihe River Catchment during 2016–2017. The investigated compounds included veterinary antibiotics, human-use antibiotics, and those intended for both human and animal use. Measurements reported from previous studies were compared with the results of this study and indicated that the use of veterinary antibiotics is probably increasing around the catchment. The ∑antibiotics concentration (i.e. the summed concentration of the 15 target antibiotics) ranged from 414 to 1951 ng/L, with an average of 821 ng/L. Discharges from wastewater treatment plants were the main sources of these compounds. The mass loading of antibiotics to the river waters was higher during wet seasons than during dry seasons. The mass fluxes of the antibiotics continuously increased towards the lower reaches of the rivers. The total annual input of the antibiotics from the Haihe River and Yongdingxin River into the Bohai Bay was 5008 kg/yr. Ofloxacin, trimethoprim, leucomycin, anhydro erythromycin and florfenicol were the predominant antibiotics, whilst amoxicillin, anhydro erythromycin, ofloxacin, norfloxacin and enrofloxacin may pose high ecotoxicological risks to the investigated aquatic ecosystem. Several antibiotics selected in this study were reported for the first time in this catchment. This study provides important information for chemical management and indicates that further monitoring is needed on the more harmful and veterinary antibiotics in the catchment. © 2019 The Authors