{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:39:33Z","timestamp":1760143173928,"version":"build-2065373602"},"reference-count":28,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2024,1,17]],"date-time":"2024-01-17T00:00:00Z","timestamp":1705449600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Metabolites"],"abstract":"<jats:p>Changes in thyroid hormone (TH) levels in rat brain at early developmental stages are correlated with adverse effects on offspring development. To characterize the ability of substances to interfere with the TH concentrations in, e.g., rat brain, it is essential to know the mean TH concentrations in this tissue under control conditions. In this publication, an online solid-phase extraction (SPE) liquid chromatography (LC) tandem mass spectrometry (MS\/MS) method was validated and used to measure TH metabolites (T4, T3, rT3, T2 and T1) in the brains of untreated rats. Data on TH concentrations in the whole brain and separate data from the cerebellum and the cortex are shown. The corresponding samples were gathered from young rats at postnatal days (PND) 4 and 21\/22 and from adult rats. The results show inter alia the high accuracy and precision of the method, and LOQs of 0.02 ng\/mL were determined for T1, T2 and rT3 and of 0.15 ng\/mL for T3 and T4. Technical variability is low, as shown by the relative standard deviations of 7.5\u201320%. For our rat model, we found that T4, T3 and T2 concentrations rise from PND4 to PND21, whereas the rT3 concentration decreases; as well as there is no statistical difference between TH concentrations in the male and female rat brain. This method is suitable to analyze TH metabolites in the brain and build up a database of historical TH concentrations in control rats. Together, this yields a robust diagnostic tool to detect potentially adverse disturbances of TH homeostasis in the most vulnerable anatomic structure.<\/jats:p>","DOI":"10.3390\/metabo14010061","type":"journal-article","created":{"date-parts":[[2024,1,17]],"date-time":"2024-01-17T07:41:28Z","timestamp":1705477288000},"page":"61","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Thyroid Hormone Metabolites Quantified in Pup and Adult Rat Cerebellum, Cortex and Whole-Brain Samples Using an Automated Online SPE-LC-MS\/MS Method"],"prefix":"10.3390","volume":"14","author":[{"given":"Christiane","family":"Hindrichs","sequence":"first","affiliation":[{"name":"BASF Metabolome Solutions GmbH, Tegeler Weg 33, 10589 Berlin, Germany"},{"name":"Department of Chemistry, Rheinland-Pf\u00e4lzischen Technischen Universit\u00e4t Kaiserslautern-Landau, Erwin-Schr\u00f6dinger-Stra\u00dfe 52, 67663 Kaiserslautern, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6341-3107","authenticated-orcid":false,"given":"Tilmann","family":"Walk","sequence":"additional","affiliation":[{"name":"BASF Metabolome Solutions GmbH, Tegeler Weg 33, 10589 Berlin, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert","family":"Landsiedel","sequence":"additional","affiliation":[{"name":"Experimental Toxicology and Ecology, BASF SE, Im Spitzenbusch 10, 67227 Frankenthal, Germany"},{"name":"Pharmacology and Toxicology, Institute of Pharmacy, Free University of Berlin, 14195 Berlin, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hennicke","family":"Kamp","sequence":"additional","affiliation":[{"name":"BASF Metabolome Solutions GmbH, Tegeler Weg 33, 10589 Berlin, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Steffen","family":"Schneider","sequence":"additional","affiliation":[{"name":"Experimental Toxicology and Ecology, BASF SE, Im Spitzenbusch 10, 67227 Frankenthal, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephanie","family":"Melching-Kollmuss","sequence":"additional","affiliation":[{"name":"Agricultural Solutions, BASF SE, Speyerer Str. 2, 67117 Limburgerhof, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dorothee","family":"Funk-Weyer","sequence":"additional","affiliation":[{"name":"Experimental Toxicology and Ecology, BASF SE, Im Spitzenbusch 10, 67227 Frankenthal, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,1,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"40","DOI":"10.3389\/fendo.2014.00040","article-title":"Thyroid hormones, T3 and T4, in the brain","volume":"5","author":"Schroeder","year":"2014","journal-title":"Front. Endocrinol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Chatonnet, F., Picou, F., Fauquier, T., and Flamant, F. (2011). Thyroid Hormone Action in Cerebellum and Cerebral Cortex Development. J. Thyroid. Res., 145762.","DOI":"10.4061\/2011\/145762"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1210\/endo-110-2-367","article-title":"An Analysis of the Sources and Quantity of 3,5,3\u2032-Triiodothyronine Specifically Bound to Nuclear Receptors in Rat Cerebral Cortex and Cerebellum","volume":"110","author":"Crantz","year":"1982","journal-title":"Endocrinology"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/S2213-8587(15)00327-7","article-title":"Association of maternal thyroid function during early pregnancy with offspring IQ and brain morphology in childhood: A population-based prospective cohort study","volume":"4","author":"Korevaar","year":"2016","journal-title":"Lancet Diabetes Endocrinol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"95001","DOI":"10.1289\/EHP5297","article-title":"Evaluating Chemicals for Thyroid Disruption: Opportunities and Challenges with In Vitro Testing and Adverse Outcome Pathway Approaches","volume":"127","author":"Noyes","year":"2019","journal-title":"Environ. Health Perspect."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1080\/10408444.2021.1910625","article-title":"Towards a science-based testing strategy to identify maternal thyroid hormone imbalance and neurodevelopmental effects in the progeny\u2014Part II: How can key events of relevant adverse outcome pathways be addressed in toxicological assessments?","volume":"51","author":"Marty","year":"2021","journal-title":"Crit. Rev. Toxicol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1789","DOI":"10.1210\/endo.143.5.8770","article-title":"Concentrations of seven iodothyronine metabolites in brain regions and the liver of the adult rat","volume":"143","author":"Pinna","year":"2002","journal-title":"Endocrinology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1021\/ac1026995","article-title":"Determination of Six Thyroid Hormones in the Brain and Thyroid Gland Using Isotope-Dilution Liquid Chromatography\/Tandem Mass Spectrometry","volume":"83","author":"Kunisue","year":"2011","journal-title":"Anal. Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1093\/toxsci\/kfad040","article-title":"Reducing uncertainties in quantitative adverse outcome pathways by analysis of thyroid hormone in the neonatal rat brain","volume":"193","author":"Ford","year":"2023","journal-title":"Toxicol. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1093\/toxsci\/kfs335","article-title":"An Animal Model of Marginal Iodine Deficiency During Development: The Thyroid Axis and Neurodevelopmental Outcome","volume":"132","author":"Gilbert","year":"2013","journal-title":"Toxicol. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"4055","DOI":"10.1210\/en.2010-0252","article-title":"Perinatal Iron and Copper Deficiencies Alter Neonatal Rat Circulating and Brain Thyroid Hormone Concentrations","volume":"151","author":"Bastian","year":"2010","journal-title":"Endocrinology"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1093\/toxsci\/kfab080","article-title":"Thyroid Disruptors: Extrathyroidal Sites of Chemical Action and Neurodevelopmental Outcome\u2014An Examination Using Triclosan and Perfluorohexane Sulfonate","volume":"183","author":"Gilbert","year":"2021","journal-title":"Toxicol. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1093\/toxsci\/kfac038","article-title":"Gestational Exposure to Perchlorate in the Rat: Thyroid Hormones in Fetal Thyroid Gland, Serum, and Brain","volume":"188","author":"Gilbert","year":"2022","journal-title":"Toxicol. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"4662","DOI":"10.1038\/s41598-019-40249-7","article-title":"A transient window of hypothyroidism alters neural progenitor cells and results in abnormal brain development","volume":"9","author":"Thomas","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1021\/acs.chemrestox.3c00152","article-title":"A Novel and Fast Online-SPE-LC-MS\/MS Method to Quantify Thyroid Hormone Metabolites in Rat Plasma","volume":"37","author":"Hindrichs","year":"2023","journal-title":"Chem. Res. Toxicol."},{"key":"ref_16","unstructured":"(2021). Guidance Document on Pesticide Analytical Methods for Risk Assessment and Post-approval Control and Monitoring Purposes (Standard No. Sante\/2020\/12830)."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"752","DOI":"10.1289\/ehp.11089","article-title":"Developmental Exposure to Perchlorate Alters Synaptic Transmission in Hippocampus of the Adult Rat","volume":"116","author":"Gilbert","year":"2008","journal-title":"Environ. Health Perspect."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1090081","DOI":"10.3389\/fendo.2023.1090081","article-title":"Thyroid hormone action controls multiple components of cell junctions at the ventricular zone in the newborn rat brain","volume":"14","author":"McMichael","year":"2023","journal-title":"Front. Endocrinol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1210\/endo.140.2.6537","article-title":"Expression profiles of the three iodothyronine deiodinases, D1, D2, and D3, in the developing rat","volume":"140","author":"Bates","year":"1999","journal-title":"Endocrinology"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/S0083-6729(05)71004-9","article-title":"Thyroid Hormones and Brain Development","volume":"71","author":"Bernal","year":"2005","journal-title":"Vitam. Horm."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1007\/s12311-008-0021-4","article-title":"Thyroid hormone and cerebellar development","volume":"7","author":"Anderson","year":"2008","journal-title":"Cerebellum"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1111\/jne.12135","article-title":"Altered cerebellum development and dopamine distribution in a rat genetic model with congenital hypothyroidism","volume":"26","author":"Shimokawa","year":"2014","journal-title":"J. Neuroendocrinol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1093\/toxsci\/kfy190","article-title":"Thyroid Hormone Disruption in the Fetal and Neonatal Rat: Predictive Hormone Measures and Bioindicators of Hormone Action in the Developing Cortex","volume":"166","author":"Wood","year":"2018","journal-title":"Toxicol. Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1157","DOI":"10.1210\/en.2013-1571","article-title":"Fetal and neonatal iron deficiency exacerbates mild thyroid hormone insufficiency effects on male thyroid hormone levels and brain thyroid hormone-responsive gene expression","volume":"155","author":"Bastian","year":"2014","journal-title":"Endocrinology"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1093\/toxsci\/kfx163","article-title":"Neurodevelopment and Thyroid Hormone Synthesis Inhibition in the Rat: Quantitative Understanding Within the Adverse Outcome Pathway Framework","volume":"160","author":"Hassan","year":"2017","journal-title":"Toxicol. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1890","DOI":"10.1210\/endo-117-5-1890","article-title":"Effects of Maternal Hypothyroidism on the Weight and Thyroid Hormone Content of Rat Embryonic Tissues, before and after Onset of Fetal Thyroid Function","volume":"117","author":"Escobar","year":"1985","journal-title":"Endocrinology"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1016\/S0021-9258(18)64849-5","article-title":"A simple method for the isolation and purification of total lipides from animal tissues","volume":"226","author":"Lees","year":"1957","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1016\/0003-9861(82)90229-6","article-title":"Preparation of lipid-free tissue extracts for chromatographic determination of thyroid hormones and metabolites","volume":"216","author":"Gordon","year":"1982","journal-title":"Arch. Biochem. Biophys."}],"container-title":["Metabolites"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-1989\/14\/1\/61\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T13:48:50Z","timestamp":1760104130000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-1989\/14\/1\/61"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,1,17]]},"references-count":28,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,1]]}},"alternative-id":["metabo14010061"],"URL":"https:\/\/doi.org\/10.3390\/metabo14010061","relation":{},"ISSN":["2218-1989"],"issn-type":[{"type":"electronic","value":"2218-1989"}],"subject":[],"published":{"date-parts":[[2024,1,17]]}}}