Li Zhong-Min, Giesert Florian, Vogt-Weisenhorn Daniela, Main Katharina Maria, Skakkebæk Niels Erik, Kiviranta Hannu, Toppari Jorma, Feldt-Rasmussen Ulla, Shen Heqing, Schramm Karl-Werner, De Angelis Meri
Helmholtz Zentrum München-German Research Center for Environmental Health (GmbH), Molecular EXposomics, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
Technische Universität München-Weihenstephan, Lehrstuhl für Entwicklungsgenetik, c/o Helmholtz Zentrum München, Ingolstädter Landstr. 1, 85764, Neuherberg, Germany.
J Chromatogr A. 2018 Jan 26;1534:85-92. doi: 10.1016/j.chroma.2017.12.048. Epub 2017 Dec 24.
The transplacental passage of thyroid hormones (THs) is of great significance since the maternal THs are vitally important in ensuring the normal fetal development. In this paper, we determined the concentrations of seven THs, viz. L-thyroxine (T), 3,3',5-triiodo-l-thyronine (T), 3,3',5'-triiodo-l-thyronine (rT), 3,3'-diiodo-l-thyronine (T), 3,5-diiodo-l-thyronine (rT), 3-iodo-l-thyronine (T) and 3-iodothyronamine (TAM), in placenta using isotope dilution liquid chromatography quadrupole time-of-flight mass spectrometry. We optimized the method using isotopically labeled quantification standards (C-T, C-T, C-rT and C-T) and recovery standard (C-T) in combination with solid-liquid extraction, liquid-liquid extraction and solid phase extraction. The linearity was obtained in the range of 0.5-150 pg uL with R values >0.99. The method detection limits (MDLs) ranged from 0.01 ng g to 0.2 ng g, while the method quantification limits (MQLs) were between 0.04 ng g and 0.7 ng g. The spike-recoveries for THs (except for T and TAM) were in the range of 81.0%-112%, with a coefficient of variation (CV) of 0.5-6.2%. The intra-day CVs and inter-day CVs were 0.5%-10.3% and 1.19%-8.88%, respectively. Concentrations of the THs were 22.9-35.0 ng g T, 0.32-0.46 ng g T, 2.86-3.69 ng g rT, 0.16-0.26 ng g T, and < MDL for other THs in five human placentas, and 2.05-3.51 ng g T, 0.37-0.62 ng g T, 0.96-1.3 ng g rT, 0.07-0.13 ng g T and < MDL for other THs in five mouse placentas. The presence of T was tracked in placenta for the first time. This method with improved selectivity and sensitivity allows comprehensive evaluation of TH homeostasis in research of metabolism and effects of environmental contaminant exposures.
甲状腺激素(THs)的胎盘转运具有重要意义,因为母体甲状腺激素对确保胎儿正常发育至关重要。在本文中,我们使用同位素稀释液相色谱-四极杆飞行时间质谱法测定了胎盘中七种甲状腺激素的浓度,即L-甲状腺素(T4)、3,3',5-三碘-L-甲状腺原氨酸(T3)、3,3',5'-三碘-L-甲状腺原氨酸(反T3)、3,3'-二碘-L-甲状腺原氨酸(3,3'-T2)、3,5-二碘-L-甲状腺原氨酸(3,5-T2)、3-碘-L-甲状腺原氨酸(3-T1)和3-碘甲状腺胺(TAM)。我们使用同位素标记的定量标准品(C-T4、C-T3、C-反T3和C-3,3'-T2)和回收率标准品(C-3,5-T2)结合固液萃取、液液萃取和固相萃取对方法进行了优化。在0.5-150 pg μL范围内获得了线性关系,R值>0.99。方法检测限(MDLs)范围为0.01 ng/g至0.2 ng/g,而方法定量限(MQLs)在0.04 ng/g至0.7 ng/g之间。甲状腺激素(T4和TAM除外)的加标回收率在81.0%-112%范围内,变异系数(CV)为0.5%-6.2%。日内CV和日间CV分别为0.5%-10.3%和1.19%-8.88%。在五个人类胎盘中,甲状腺激素的浓度分别为22.9-35.0 ng/g T4、0.32-0.46 ng/g T3、2.86-3.69 ng/g反T3、0.16-0.26 ng/g 3,3'-T2,其他甲状腺激素低于MDL;在五个小鼠胎盘中,甲状腺激素的浓度分别为2.05-3.51 ng/g T4、0.37-0.62 ng/g T3、0.96-1.3 ng/g反T3、0.07-0.13 ng/g 3,3'-T2,其他甲状腺激素低于MDL。首次在胎盘中追踪到了3,5-T2的存在。这种具有更高选择性和灵敏度的方法能够在环境污染物暴露的代谢和影响研究中对甲状腺激素稳态进行全面评估。