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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
- Source :
- Frontiers in Endocrinology, Vol 10 (2019), Frontiers in Endocrinology
- Publication Year :
- 2019
- Publisher :
- Frontiers Media SA, 2019.
-
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.
- Subjects :
- 0301 basic medicine
thyroid hormones metabolites
Endocrinology, Diabetes and Metabolism
030209 endocrinology & metabolism
Endogeny
Derivative
Mass spectrometry
lcsh:Diseases of the endocrine glands. Clinical endocrinology
High-performance liquid chromatography
03 medical and health sciences
chemistry.chemical_compound
Endocrinology
0302 clinical medicine
Solid phase extraction
Original Research
mass spectrometry
thyroid hormones
T2
lcsh:RC648-665
Chromatography
3, 5-diiodo-L-thyronine
Thyroid Hormones
3,5-Diiodo-L-Thyronine
Mass Spectrometry
Hexane
030104 developmental biology
chemistry
Thyronine
5-Diiodo-L-Thyronine
Hormone
Subjects
Details
- ISSN :
- 16642392
- Volume :
- 10
- Database :
- OpenAIRE
- Journal :
- Frontiers in Endocrinology
- Accession number :
- edsair.doi.dedup.....1be91b9fa04e6bb757d96547b634e488