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Cited by 22Thyroid Disorders and TreatmentsAnalytical chemistry methods developmentAnalytical Methods in Pharmaceuticals

Assay of endogenous 3,5-diiodo-L-thyronine (3,5-T2) and 3,3'-diiodo-L-thyronine (3,3'-T2) in human serum: a feasibility study

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Lorenzini, Leonardo, Nguyen, Nhat Minh, Sacripanti, Ginevra, Serni, Enrico, Borsò, Marco, Saponaro, Federica, Cecchi, Elena, Simoncini, Tommaso +3 more·2019·Frontiers in Endocrinology
DOI 10.3389/fendo.2019.00088PMC PMC6389606
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Abstract

3,5-diiodo-L-thyronine (3,5-T2) is an endogenous derivative of thyroid hormone with potential metabolic effects. It has been detected in human blood by immunological methods, but a reliable assay based on mass spectrometry (MS), which is now regarded as the gold standard in clinical chemistry, is not available yet. Therefore, we aimed at developing a novel ad-hoc optimized method to quantitate 3,5-T2 and its isomers by MS in human serum. Serum samples were obtained from 28 healthy subjects. Two ml of serum were deproteinized with acetonitrile and then subjected to an optimized solid phase extraction-based procedure. To lower background noise, the samples were furtherly cleaned by hexane washing and acetonitrile precipitation of residual proteins. 3,5-T2 and its isomers 3,3’-T2 and 3’,5’-T2 were then analyzed by HPLC coupled to tandem MS. Accuracy and precision for T2 assay were 88-104% and 95-97%, respectively. Recovery and matrix effect averaged 78% and +8%, respectively. 3,5-T2 was detected in all samples and its concentration averaged (mean±SEM) 41±5 pg/ml, i.e. 78±9 pmol/l. In the same samples the concentration of 3,3’-T2 averaged 133±15 pg/ml, i.e. 253±29 pmol/l, while 3’,5’-T2 was not detected. 3,5-T2 concentration was significantly related to 3,3’-T2 concentration (r=0.540, P<0.01), while no significant correlation was observed with either T3 or T4 in a subset of patients in which these hormones were assayed. In conclusion, our method is able to quantify 3,5-T2 and 3,3’-T2 in human serum. Their concentrations lie in the subnanomolar range, and a significant correlation was detected between these two metabolites in healthy individuals.

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  • The higher LC-MS-based 3,5-T2 concentrations reported by Lorenzini et al. are probably due to contamination of the 13C9-15N-3,5-T2 internal standard with natural unlabeled 3,5-T2.

    “The fact that Lorenzini et al. probably report higher LC-MS-based 3,5-T2 concentrations is due to the contamination of the 13C9-15N-3,5-T2 internal standard, with “natural” unlabeled 3,5-T2.”

    From 3,5-T2—A Janus-Faced Thyroid Hormone Metabolite Exerts Both Canonical T3-Mimetic Endocrine and Intracrine Hepatic Action (2020) · cites this paper as [75]

    The Results report LC-MS-based 3,5-T2 measurements, and Materials and Methods (Reagents and Materials; Sample Preparation) identify 13C9¹⁵N-T2 as an internal standard. Neither section reports contamination with unlabeled 3,5-T2 or attributes the measured concentrations to it; that causal explanation is unsupported by this paper. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • Lorenzini et al. observed a mean 3,5-T2 concentration in human serum of 78 ± 9 pM by LC-MS.

    “Lorenzini et al. (75), observed a mean concentration range for 3,5-T2 in human serum of 78 ± 9 pM”

    From 3,5-T2—A Janus-Faced Thyroid Hormone Metabolite Exerts Both Canonical T3-Mimetic Endocrine and Intracrine Hepatic Action (2020) · cites this paper as [75]

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Research works printed but not linked: 22

  1. [1]Accorroni A Saponaro F Zucchi R. Tissue thyroid hormones and thyronamines. Heart Fail Rev. (2016) 21: 373– 90. 10.1007/s10741-016-9553-8 27115768 · DOI · Paper page
  2. [2]Senese R Cioffi F De Lange P Goglia F Lanni A. Thyroid: biological actions of “nonclassical” thyroid hormones. J Endocrinol. (2014) 221: R1– 12. 10.1530/JOE-13-0573 24464019 · DOI
  3. [3]Moreno M Lanni A Lombardi A Goglia F. How the thyroid controls metabolism in the rat: different roles for triiodothyronine and diiodothyronines. J Physiol. (1997) 505 (Pt. 2): 529– 38. 9423191 PMC1160082
  4. [4]Meinhold H Shurnbarnd P. A radioimmunoassay for 3,5-diiodothyronine*. Clin Endocrinol. (1978) 8: 493– 7. 10.1111/j.1365-2265.1978.tb02187.x 668156 · DOI
  5. [5]Faber J Lumholtz IB Kirkegaard C Siersbaek-Nielsen KFT. Isolation of radioactive iodothyronines for kinetic studies: a comparison of two methods. Acta Endocrinol. (1982) 99: 64– 71. 10.1530/acta.0.0990064 7058678 · DOI
  6. [6]Pinna G Meinhold H Hiedra L Thoma R Hoell T Gräf KJ. Elevated 3,5-diiodothyronine concentrations in the sera of patients with nonthyroidal illnesses and brain tumors. J Clin Endocrinol Metab. (1997) 82: 1535– 42. 9141546
  7. [7]Lehmphul I Brabant G Wallaschofski H Ruchala M Strasburger CJ Koehrle J. Detection of 3,5-diiodothyronine in sera of patients with altered thyroid status using a new monoclonal antibody based chemiluminescence immunoassay. Thyroid (2014) 24: 1350– 60. 10.1089/thy.2013.0688 24967815 · DOI · Paper page
  8. [8]Wang D Stapleton HM. Analysis of thyroid hormones in serum by liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem. (2010) 397: 1831– 9. 10.1007/s00216-010-3705-9 20437035 PMC3082288 · DOI
  9. [9]Saba A Chiellini G Frascarelli S Marchini M Ghelardoni S Raffaelli A. Tissue distribution and cardiac metabolism of 3-iodothyronamine. Endocrinology (2010) 151: 5063– 73. 10.1210/en.2010-0491 20739399 · DOI
  10. [10]Galli E Marchini M Saba A Berti S Tonacchera M Vitti P. Detection of 3-iodothyronamine in human patients: a preliminary study. J Clin Endocrinol Metab. (2012) 97: E69– 74. 10.1210/jc.2011-1115 22031514 · DOI
  11. [11]Saba A Donzelli R Colligiani D Raffaelli A Nannipieri M Kusmic C. Quantification of thyroxine and 3,5,3′-triiodo-thyronine in human and animal hearts by a novel liquid chromatography-tandem mass spectrometry method. Horm Metab Res. (2014) 46: 628– 34. 10.1055/s-0034-1368717 24591048 · DOI
  12. [12]Matuszewski BK Constanzer ML Chavez-Eng CM. Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS. Anal Chem. (2003) 75: 3019– 30. 10.1021/ac020361s 12964746 · DOI
  13. [13]Pietzner M Lehmphul I Friedrich N Schurmann C Ittermann T Dörr M. Translating pharmacological findings from hypothyroid rodents to euthyroid humans: is there a functional role of endogenous 3,5-T2? Thyroid (2015) 25: 188– 97. 10.1089/thy.2014.0262 25343227 PMC4322092 · DOI · Paper page
  14. [14]Pietzner M Homuth G Budde K Lehmphul I Völker U Völzke H Urine Metabolomics by &lt;sup&gt;1&lt;/sup&gt;H-NMR spectroscopy indicates associations between serum 3,5-T&lt;sub&gt;2&lt;/sub&gt; concentrations and intermediary metabolism in euthyroid humans. Eur Thyroid J. (2015) 4 (Suppl. 1): 92– 100. 10.1159/000381308 26601079 PMC4640298 · DOI
  15. [15]Soldin OP Soldin SJ. Thyroid hormone testing by tandem mass spectrometry. Clin Biochem. (2011) 44: 89– 94. 10.1016/j.clinbiochem.2010.07.020. 20691174 PMC3634329 · DOI
  16. [16]Carvalho VM. The coming of age of liquid chromatography coupled to tandem mass spectrometry in the endocrinology laboratory. J Chromatogr B Anal Technol Biomed Life Sci. (2012) 883–884: 50– 8. 10.1016/j.jchromb.2011.08.027 21907642 · DOI · Paper page
  17. [17]Richards K Rijntjes E Rathmann D Köhrle J Avoiding the pitfalls when quantifying thyroid hormones and their metabolites using mass spectrometric methods: the role of quality assurance. Mol Cell Endocrinol. (2017) 58: 44– 56. 10.1016/j.mce.2017.01.032 28153800 · DOI · Paper page
  18. [18]Donzelli R Colligiani D Kusmic C Sabatini M Lorenzini L Accorroni A. Effect of hypothyroidism and hyperthyroidism on tissue thyroid hormone concentrations in rat. Eur Thyroid J. (2016) 5: 27– 34. 10.1159/000443523 27099836 PMC4836127 · DOI
  19. [19]Jonklaas J Sathasivam A Wang H Finigan D Soldin O Burman K. 3,3′-Diiodothyronine concentrations in hospitalized or thyroidectomized patients: results from a pilot study. Endocr Pract. (2014) 20: 797– 807. 10.4158/EP13453.OR 24518182 PMC4185286 · DOI · Paper page
  20. [20]Hansen M Luong X Sedlak DL Helbing CC Hayes T. Quantification of 11 thyroid hormones and associated metabolites in blood using isotope-dilution liquid chromatography tandem mass spectrometry. Anal Bioanal Chem. (2016) 408: 5429– 42. 10.1007/s00216-016-9614-9 27215639 · DOI
  21. [21]Li ZM Giesert F Vogt-Weisenhorn D Main KM Skakkebæk NE Kiviranta H. Determination of thyroid hormones in placenta using isotope-dilution liquid chromatography quadrupole time-of-flight mass spectrometry. J Chromatogr A (2018) 1534: 85– 92. 10.1016/j.chroma.2017.12.048 29307529 · DOI
  22. [22]Lorenzini L Ghelardoni S Saba A Sacripanti G Chiellini G Zucchi R. Recovery of 3-iodothyronamine and derivatives in biological matrixes: problems and pitfalls. Thyroid (2017) 27: 1323– 31. 10.1089/thy.2017.0111 28859548 · DOI

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Authors
Lorenzini, L., Nguyen, N. M., Sacripanti, G., Serni, E., Borsò, M., Saponaro, F., Cecchi, E., Simoncini, T., Ghelardoni, S., Zucchi, R., & Saba, A.
Affiliations
University of Pisa (IT)
Corresponding author
  • Saponaro, Federica
  • Zucchi, Riccardo
  • Saba, Alessandro
ORCID
Venue
Frontiers in Endocrinology
Publisher
Frontiers Media SA
Published
2019-02-19
Volume
10
Pages
88-88
ISSN
1664-2392 (ISSN-L)
DOI
10.3389/fendo.2019.00088
PMC
PMC6389606
OpenAlex
W2917149508
Cited by
22
References
0
Citations by year
2025: 1 · 2024: 3 · 2022: 5 · 2021: 1 · 2020: 10 · 2019: 2
FWCI
3.65
Type
journal article
Language
en
Open access
Licence
Locations
  • Frontiers in Endocrinology (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • CINECA IRIS Institutial research information system (University of Pisa) (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
Università di Pisa
Topics
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Thyroid Disorders and Treatments
  • Physical Sciences › Chemistry › Analytical Chemistry › Analytical chemistry methods development
  • Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
Keywords
3,5-diiodo-L-thyronine, thyroid hormone metabolites, solid-phase extraction, matrix effects, healthy volunteers

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