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This study aimed to investigate the effect of dissolved humic acid (dHA) on Zn bioavailability and the subsequent influence on the Zn-induced thyroid toxicity. Zn toxicity was assessed using a yeast bioassay in the presence and absence of dHA. With increasing concentration of dHA, the toxic effects decreased, and the free Zn concentrations detected by the anodic stripping voltammetry (ASV) method also decreased. The high correlation (R = 0.92, p < 0.001) between toxic effects and free Zn concentrations indicated that Zn thyroid toxicity largely comes from the free Zn fraction. Water samples from the Qing River in Beijing were also assayed for thyroid toxicity. The results revealed that the metals might contribute to the toxicity. The known thyroid hormone-disrupting metals, namely, Zn, Cd and Hg, were analyzed. The cause-effect relationship between the observed thyroid toxicity and free Zn concentrations as well as their dose-effect relationships were examined. Our results showed that Zn might be the major contributor to the observed thyroid toxicity caused by metals. These results suggest that the ASV method and the identified major contributor (Zn) may be used in lieu of conventional environmental analyses to follow the progression of a risk assessment or remediation strategy., (Copyright © 2018 Elsevier Inc. All rights reserved.)