Quantification of 11 thyroid hormones and associated metabolites in blood using isotope-dilution liquid chromatography tandem mass spectrometry
Quantification of 11 thyroid hormones and associated metabolites in blood using isotope-dilution liquid chromatography tandem mass spectrometry
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DOI：
10.1007/s00216-016-9614-9
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发表时间：
2016-08-01
影响因子：
4.3
通讯作者：
Hayes, Tyrone
中科院分区：
文献类型：
--
作者：
Hansen, Martin;Luong, Xuan;Hayes, Tyrone
本文介绍了一种同时测定甲状腺素（T-4）、3，3 '，5-三碘甲状腺原氨酸（T-3）、3，3'，5 '-三碘甲状腺原氨酸（rT（3））、3，5-二碘甲状腺原氨酸（3，5-T-2）、3，3'-二碘甲状腺原氨酸（3，3 '-diiodothyronine，3，3'-diiodothyronine，3，3 '-diiodothyronine，rT（3））等11种天然甲状腺激素及其代谢产物的绝对浓度和总浓度的新分析方法（3，3 '-T-2）、3-碘甲状腺原氨酸（T-1）、甲状腺原氨酸（T-0）、3-碘甲状腺原氨酸（T（1）AM）、四碘甲状腺乙酸（Tetrac）、三碘甲状腺乙酸（Triac）和二碘甲状腺乙酸（Diac）。使用四种同位素标记的替代品和内标物结合固相萃取和LC-MS/MS优化了该方法。使用两栖动物血浆和血清与基质匹配校准进行定量，进一步评估了该方法。方法检测限为3.5 pg T-4、1.5 pg T-3、2.9 pg rT（3）、1.7 pg 3，3 '-T-2、2.3 pg 3，5-T-2，其余6种代谢物在50 μ L等份血清或血浆中的检测限为0.3 - 7.5 pg。所有分析物的准确度和重复性分别为88 - 103%和1.31 - 17.2%。最后，我们将该方法应用于成年青蛙（非洲爪蟾）血浆和蝌蚪（蛙（Lithobates）catesbeiana）血清。我们在蝌蚪血清中观察到多达七种不同的甲状腺激素和相关代谢物。这种方法将使研究人员能够改善动物和人类甲状腺稳态和内分泌干扰的评估。
This paper describes a novel analytical methodology for the simultaneous determination of absolute and total concentrations of 11 native thyroid hormones and associated metabolites, viz. thyroxine (T-4), 3,3', 5-triiodothyronine (T-3), 3,3', 5'-triiodothyronine (rT(3)), 3,5-diiodothyronine (3,5-T-2), 3,3'- diiodothyronine (3,3'-T-2), 3-iodothyronine (T-1), thyronine (T-0), 3-iodothyronamine (T(1)AM), tetraiodothyroacetic acid (Tetrac), triiodothyroacetic acid (Triac), and diiodothyroacetic acid (Diac), in 50-mu L of plasma or serum. The method was optimized using four isotopic labeled surrogate and internal standards in combination with solid-phase extraction and LC-MS/MS. The methodology was further evaluated using amphibian plasma and serum with matrix-matched calibration applied for quantification. Method detection limits are 3.5 pg T-4, 1.5 pg T-3, 2.9 pg rT(3), 1.7 pg 3,3'-T-2, 2.3 pg 3,5-T-2, and between 0.3 and 7.5 pg for the remaining six metabolites in 50 mu L aliquots of blood sera or plasma. Accuracies and repeatabilities for all analytes were between 88 and 103 % and 1.31 and 17.2 %, respectively. Finally, we applied the method on adult frog (Xenopus laevis) plasma and tadpole (Rana (Lithobates) catesbeiana) serum. We observed up to seven different thyroid hormones and associated metabolites in tadpole serum. This method will enable researchers to improve the assessment of thyroid homeostasis and endocrine disruption in animals and humans.