Bisphenols (BPs), which act as endocrine disruptors, are extensively used in epoxy resin and polycarbonate plastic production and are found in various environmental media. This study investigated the occurrence and distribution of seven BPs in water and sediment samples from the Lanzhou section of the Yellow River, China. Previous studies have focused on the southern and more developed regions in China. Paired water and sediments were collected in the Yellow River from August 2016 to March 2017. Bisphenol A (BPA), bisphenol AF (BPAF) and bisphenol S (BPS) were measured in the study samples. Mean concentrations of BPs were 88 and 61.2 ng/L in the rainy and dry seasons, respectively, in water samples. The concentrations were 40.4 and 29.9 ng/g in the rainy and dry seasons, respectively, in sediment samples. BPA accounted for the largest proportion in ∑BPs, followed by BPAF and BPS, which indicates that BPA and BPAF have been widely used and discharged in this region. 17β-estradiol equivalents for BP analogues were 21.57–203.83 pg/L and 13.06–185.30 pg/L in the dry and rainy seasons, respectively, in water samples. BPAF had the highest mean estrogenic activity (> 80%) amongst the study samples. The concentration of ∑BPs was lower than those in previous studies; however, the concentration of BPAF was high. The present study also surveyed the estrogenicity of BPs from the samples, and BPAF contributed the highest mean estrogenic activity. The results of this study demonstrate that BP pollution exists to some degree, and the use and discharge of BPAF should be monitored.