Horm Metab Res 2014; 46(09): 628-634
DOI: 10.1055/s-0034-1368717
Endocrine Research
© Georg Thieme Verlag KG Stuttgart · New York

Quantification of Thyroxine and 3,5,3′-Triiodo-Thyronine in Human and Animal Hearts by a Novel Liquid Chromatography-Tandem Mass Spectrometry Method

A. Saba
1   Department of Pathology, University of Pisa, Pisa, Italy
,
R. Donzelli
1   Department of Pathology, University of Pisa, Pisa, Italy
,
D. Colligiani
2   Department of Medicine, University of Pisa, Pisa, Italy
,
A. Raffaelli
3   National Research Council, Institute of Clinical Physiology, Pisa, Italy
,
M. Nannipieri
2   Department of Medicine, University of Pisa, Pisa, Italy
,
C. Kusmic
3   National Research Council, Institute of Clinical Physiology, Pisa, Italy
,
C. G. Dos Remedios
4   Muscle Research Unit, Bosch Institute, The University of Sydney, Sydney, Australia
,
W. S. Simonides
5   Laboratory for Physiology, Institute for Cardiovascular Research, VU ­University Medical Center, Amsterdam, The Netherlands
,
G. Iervasi
3   National Research Council, Institute of Clinical Physiology, Pisa, Italy
6   G. Monasterio Foundation, Pisa, Italy
,
R. Zucchi
1   Department of Pathology, University of Pisa, Pisa, Italy
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Publikationsverlauf

received 17. August 2013

accepted 27. Januar 2014

Publikationsdatum:
03. März 2014 (online)

Abstract

Assaying tissue T3 and T4 would provide important information in experimental and clinical investigations. A novel method to determine tissue T3 and T4 by HPLC coupled to mass spectrometry is described. The major difference vs. previously described methods lies in the addition of a derivatization step, that is, to convert T3 and T4 into the corresponding butyl esters. The yield of esterification was ̴ 100% for T3 and 80% for T4. The assay was linear (r>0.99) in the range of 0.2–50 ng/ml, accuracy was in the order of 70–75%, and the minimum tissue amount needed was in the order of 50 mg, that is, about one order of magnitude lower than observed with the same equipment (AB Sciex API 4000 triple quadrupole mass spectrometer) if derivatization was omitted. The method allowed detection of T3 and T4 in human left ventricle biopsies yielding concentrations of 1.51±0.16 and 5.94±0.63 pmol/g, respectively. In rats treated with different dosages of exogenous T3 or T4, good correlations (r>0.90) between plasma and myocardial T3 and T4 concentrations were observed, although in specific subsets different plasma T4 concentrations were not associated with different tissue content in T4. We conclude that this method could provide a novel insight into the relationship between plasma and tissue thyroid hormone levels.

 
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