Simultaneous quantification of T4, T3, rT3, 3,5‐T2 and 3,3′‐T2 in larval zebrafish (Danio rerio) as a model to study exposure to polychlorinated biphenyls

0301 basic medicine 570 Thyroid Hormones Endocrine Disruptors 01 natural sciences Analytical Chemistry 03 medical and health sciences Limit of Detection Tandem Mass Spectrometry 615 derivatization Animals PCBs Chromatography, High Pressure Liquid Zebrafish thyroid hormones Chromatography Biomedical and Clinical Sciences Animal Reproducibility of Results Pharmacology and Pharmaceutical Sciences zebrafish Polychlorinated Biphenyls 0104 chemical sciences Disease Models, Animal Pharmacology and pharmaceutical sciences UPLC-MS/MS High Pressure Liquid Larva Chemical Sciences Disease Models Linear Models Analytical chemistry
DOI: 10.1002/bmc.4185 Publication Date: 2018-01-04T00:32:48Z
ABSTRACT
AbstractEnvironmental toxicants that interfere with thyroid hormone (TH) signaling can impact growth and development in animals and humans. Zebrafish represent a model to study chemically induced TH disruption, prompting the need for sensitive detection of THs. Simultaneous quantification of 3,3′,5‐triiodo‐l‐thyronine (T3), thyroxine (T4), 3,3′,5′‐triiodo‐l‐thyronine (rT3), 3,5‐diiodo‐l‐thyronine (3,5‐T2) and 3,3′‐diiodo‐l‐thyronine (3,3′‐T2) in zebrafish larvae was achieved by ultra‐performance liquid chromatography–tandem mass spectrometry in positive ion mode. Solid‐phase extraction with SampliQ cartridges and derivatization with 3 m hydrochloric acid in n‐butanol reduced matrix effects. Derivatized compounds were separated on an Acquity UPLC BEH C18 column with mobile phases consisting of 0.1% acetic acid in deionized water and 0.1% acetic acid in methanol. The limits of detection ranged from 0.5 to 0.6 pg injected on column. The method was validated by evaluating recovery (77.1–117.2%), accuracy (87.3–123.9%) and precision (0.5–12.4%) using diluted homogenized zebrafish embryos spiked with all target compounds. This method was then applied to zebrafish larvae collected after 114 h of exposure to polychlorinated biphenyls (PCBs), including PCB 28, PCB 66 and PCB 95, or the technical mixture Aroclor 1254. Exposure to PCB 28 and PCB 95 increased the T4:T3 ratio and decreased the T3:rT3 ratio, demonstrating that this method can effectively detect PCB‐induced alterations in THs.
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