The enrichment and purification of hexabromocyclododecanes and its effects on thyroid in zebrafish

Abstract

Hexabromocyclododecanes (HBCDs) are the third most highly produced brominated flame retardants (BFRs) all over the world. Based on the current research status of HBCDs, zebrafish were exposed to three dietary concentrations of HBCDs (0, 10, 100, 400 ng/g) for 56 days, and followed by clean food for 28 days. In order to investigate the enrichment and purification of HBCDs in zebrafish, HBCD enantiomers in zebrafish were determined using ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). To investigate the effects of long-term exposure of HBCDs on thyroid dysfunction and oxidative stress in zebrafish, the concentrations of thyroid hormone (T3, T4, FT3 and FT4) and the activities of malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) were measured. RT-PCR was used to reveal the molecular mechanism of HBCDs' influence on thyroid hormone in zebrafish. The result of UPLC-MS/MS showed that there were three main reasons for the existence of α-HBCD as the major isomer in the organism. HBCDs had significant inhibitory effect on T3 and T4 in liver of adult zebrafish after 56 days’ exposure. Compared with the control group, the ratio of T3 and T4was significantly higher in the medium and high concentration group. The content of FT3 and FT4 in the liver tissue of zebrafish increased first and then decreased with the increase of exposure concentration. With the increase of exposure concentration, the content of MDA in zebrafish liver decreased firstly and then increased. The activity of SOD and CAT in zebrafish liver showed the opposite trend with MDA. And the concentration of GSH in liver decreased gradually, which showed a significant dose-effect relationship. HBCDs exposure has an inhibitory effect on thyroid hormone receptor gene (TRβ) and adrenocorticotropin-releasing hormone gene (Crh) in zebrafish.