ECE2018 Poster Presentations: Thyroid Endocrine Disruptors (2 abstracts)
A validated LC-Q-TOF-MS method for quantitative analysis of thyroxine and metabolites in placenta
Zhong-Min Li 1 , Florian Giesert 2 , Daniela Vogt-Weisenhorn 2, , Katharina Main 4 , Niels Skakkebæk 4 , Hannu Kiviranta 5 , Jorma Toppari 4, , Ulla Feldt-Rasmussen 7 , Heqing Shen 8 , Karl-Werner Schramm 1, & Meri De Angelis 1
1Helmholtz Zentrum München-German Research Center for Environmental Health (GmbH), Molecular EXposomics, Munich, Germany; 2Technische Universität München-Weihenstephan, Lehrstuhl für Entwicklungsgenetik, c/o Helmholtz Zentrum München, Munich, Germany; 3Helmholtz Zentrum München-German Research Center for Environmental Health (GmbH), Institute of Developmental Genetics, Munich, Germany; 4Department of Growth and Reproduction and EDMaRC, Rigshospitalet, University of Copenhagen, Copenhagen, Denmark; 5National Public Health Institute, Department of Health Security, Kuopio, Finland; 6Institute of Biomedicine, University of Turku, and Department of Paediatrics, Turku University Hospital, Turku, Finland; 7Department of Medical Endocrinology PE, Rigshospitalet, University of Copenhagen, Copenhagen, Denmark; 8Key Lab of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen, China; 9Department für Biowissenschaftliche Grundlagen, Technische Universität München, Munich, Germany.
Thyroid hormones (TH) of maternal origin are critical for the proper fetal development, especially during early pregnancy. Even minor changes in maternal TH circulation can lead to various adverse outcomes. Recent studies found that the metabolites of thyroxine (T4) also play an important physiological role. For example, 3,5-diiodo-L-thyronine (T2) and 3,3′-diiodo-L-thyronine (rT2) can suppress the thyroid stimulating hormone (TSH) level and increase the resting metabolic rate. 3-iodothyronamine (T1AM) administration in mice leads to a hypometabolic state. These metabolites may have influences on the fetus. Having the capacity to make a comprehensive analysis of T4 and the metabolites in placenta provides a diagnostic tool for the placental TH homeostasis. Routine TH assessment has long been achieved by measuring T4, triiodo-L-thyronine (T3), and TSH in blood using immunoassay (IA) method, which is of high sensitivity, but is prone to nonspecific interferences. Methods based on liquid chromatography-mass spectrometry (LC-MS) and tandem mass spectrometry (LC-MS/MS) showed better accuracy and reliability. In this study, we report a method for the determination of T4, T3, 3,3′,5′-triiodo-L-thyronine (rT3), T2, rT2, 3-iodo-L-thyronine (T1), and T1AM in the placenta. The method was optimized using isotope (13C-T4, 13C-T3, 13C-rT3, 13C-T2) dilution methodology and determined by liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS). The calibration ranges from 0.5 to 150 ng/ml with R2 values > 0.99. The method detection limits (MDLs) and the method quantification limits (MQLs) were 0.01–0.2 ng/g and 0.04–0.7 ng/g, respectively. The spike-recoveries for THs (except for T1 and T1AM) were between 81.0% and 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. The method was adopted for TH measurement in human and mouse placenta. The concentrations of T4, T3, rT3, and T2 were 22.9–35.0 ng/g, 0.32–0.46 ng/g, 2.86–3.69 ng/g, and 0.16–0.26 ng/g in human placenta, and 2.05–3.51 ng/g, 0.37–0.62 ng/g, 0.96–1.3 ng/g, and 0.07–0.13 ng/g in mouse placenta, respectively. The presence of T2 was tracked in placenta tissue for the first time, indicating improved selectivity and sensitivity of our method. The validated method allows comprehensive evaluation of total T4 and metabolites in the placenta.
Volume 56
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Endocrine Abstracts
ISSN 1470-3947 (print) | ISSN 1479-6848 (online)
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