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Cited by 38Thyroid Disorders and TreatmentsAnalytical Chemistry and ChromatographyAnalytical Methods in Pharmaceuticals

A direct comparison of liquid chromatography-mass spectrometry with clinical routine testing immunoassay methods for the detection and quantification of thyroid hormones in blood serum

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Bowerbank, Samantha L., Carlin, Michelle G., Dean, John R.·2019·Analytical and Bioanalytical Chemistry
DOI 10.1007/s00216-019-01724-2PMC PMC6522465
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Abstract

A new and improved method was developed for the determination and quantification of four "free" thyroid hormones (i.e. 3,5-diiodothyronine (T2), 3,3',5-triiodothyronine (T3), 3,3',5'-triiodothyrone (rT3) and 3,5,3',5'-tetraiodothyronine (T4)) in human serum by low- and high-resolution liquid chromatography-mass spectrometry (LC-MS). Several sample preparation strategies were investigated to obtain matrix-independent results. These strategies included solid phase extraction and matrix dilution. The developed analytical methods were then directly compared, in a blind study using patient-derived human blood serum samples, to the current clinical routine testing methods, i.e. electrochemiluminescence immunoassay and enzyme-linked immunosorbent assay. Chromatographic separation was achieved on a pentafluorophenyl (F5) column with an isocratic method of 30% aqueous phase, 70% organic phase where mobile phase A is 0.1% formic acid in water (pH 4) and mobile phase B is 0.1% formic acid in methanol (pH 4) (v/v). The high-resolution LC-MS was able to give a significant improvement in sensitivity with limits of quantification of 0.002 to 0.008 pmol/L for all four "free" thyroid hormones, as well as reduced sample preparation, making this the preferred method. However, the increase in capital cost may be beyond the capabilities of some laboratories. The LC-MS methods allow for the analysis of "free" thyroid hormones to be carried out in a significantly reduced analysis time. Clinical sample analysis showed that there was no statistical difference between the results obtained by ECLIA/ELISA and both LC-MS methods. Graphical abstract.

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  • Several laboratories have developed procedures to accurately determine total T4 and total T3 in human plasma or serum and largely agree on concentration ranges, but this consensus has not been achieved for most other TH metabolites.

    “While meanwhile several laboratories developed procedures to accurately determine total T4 and total T3 in human plasma or serum and largely agree on concentrations ranges (25, 67–73), this consensus has not been achieved yet for most of the other TH metabolites, which occur at much lower concentrations.”

    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 [69]

    The Abstract describes a procedure for measuring T3 and T4 in human serum, and Results and discussion reports strong agreement between that method and routine immunoassays. But the paper measures *free*, not total, hormones and does not establish that multiple laboratories agree on concentration ranges for total T3 and T4. The citation is attached to this first clause, not the subsequent statement about other metabolites. (LLM, conf=0.82)

    Experimental — accuracy not yet measured

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Research works printed but not linked: 27

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Cited by

Sorted by citations

Authors
Bowerbank, S. L., Carlin, M. G., & Dean, J. R.
Affiliations
Northumbria University (GB)
Corresponding author
Dean, John R.
ORCID
Venue
Analytical and Bioanalytical Chemistry
Publisher
Springer Science and Business Media LLC
Published
2019-05
Volume
411
Issue
13
Pages
2839-2853
ISSN
  • 1618-2642 (ISSN-L)
  • 1618-2650
DOI
10.1007/s00216-019-01724-2
PMC
PMC6522465
OpenAlex
W2944288092
Cited by
38
References
1
Citations by year
2026: 5 · 2024: 7 · 2023: 6 · 2022: 3 · 2021: 9 · 2020: 8
FWCI
4.06
Type
journal article
Language
en
Open access
Licence
Locations
  • Analytical and Bioanalytical Chemistry (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Northumbria Research Link (Northumbria University) (acceptedVersion)
Funders
Northumbria University
Topics
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Thyroid Disorders and Treatments
  • Physical Sciences › Chemistry › Spectroscopy › Analytical Chemistry and Chromatography
  • Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
Keywords
free thyroid hormones, liquid chromatography-mass spectrometry, immunoassay comparison, electrochemiluminescence immunoassay, enzyme-linked immunosorbent assay, 3,5-diiodothyronine, triiodothyronine, reverse T3, thyroxine, solid-phase extraction, method validation
MeSH terms
Animals, Chromatography, High Pressure Liquid, Horses, Humans, Immunoassay, Mass Spectrometry, Thyroid Hormones, Limit of Detection

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