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
AUTHORS (5)
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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