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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
Author(s) -
Chen Xiaopeng,
Walter Kyla M.,
Miller Galen W.,
Lein Pamela J.,
Puschner Birgit
Publication year - 2018
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.4185
Subject(s) - chemistry , chromatography , acetic acid , derivatization , zebrafish , danio , thyronine , detection limit , tandem mass spectrometry , high performance liquid chromatography , triiodothyronine , mass spectrometry , hormone , biochemistry , gene
Environmental 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 C 18 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.