Avoiding the pitfalls when quantifying thyroid hormones and their metabolites using mass spectrometric methods: The role of quality assurance
Section snippets
Scope of the review
The use of mass spectrometry (MS) for the analysis of thyroid hormones.
(TH) and TH metabolites (THM) in biological samples has become widespread. MS methods for the determination of TH and THM in human (Siekmann, 1987, Thienpont et al., 1994, De Brabandere et al., 1998, Tai et al., 2002, Tai et al., 2004, Thienpont et al., 1999, Van Uytfanghe et al., 2004, Soukhova et al., 2004, Gu et al., 2007, Yue et al., 2008, Saba et al., 2010, DeBarber et al., 2008, Galli et al., 2012, Jonklaas et al., 2014
Background
THM are a group of low molecular mass iodine-containing hormonally active compounds derived from the amino acid L-tyrosine. The pro-hormone L-thyroxine (T4, Fig. 1A), the more biologically active 3,3′,5-Triiodo-L-thyronine (T3, Fig. 1B) and the di-iodometabolites 3,5-T2 and 3,3′-T2 (Fig. 1D and E respectively) belong to the thyronine family of compounds. The de-carboxylated, de-iodinated derivatives of T4, include 3-iodothyronamine (3-T1AM, Fig. 1C); these are known as thyronamines.
Endogenous
The history of mass spectrometry in thyroid hormone analysis
T4 and T3 are 99.97 and 99.7% respectively bound to carrier proteins in the circulation (Spencer, 2000). The availability of TH to cells is considered to be dependent on their free rather than protein-bound concentrations (Cheng et al., 2010), making the concentrations of free T3 and T4 of major diagnostic interest to clinicians. Hence, free T4 and T3 concentrations are clinically important parameters that are routinely measured on a high throughput daily basis in blood samples from adult
Quality assurance and method validation
QA is a term associated with analytical chemistry, but less so with basic research. Whilst it is unreasonable to expect thyroid hormone researchers to apply full quality management procedures to their laboratory environment, it is appropriate to expect an adequate standard of QA to be applied to published bioanalytical MS methods. What should such a standard include? The US Food and Drug Administration (FDA) guidelines for bioanalytical method validation (B.C. Committee, 2001) state that
Immunoassays
In the decades before the introduction of the LC-MS/MS methodology, the thyroid field depended on the use of IA for the quantification of TH (Wiersinga and Chopra, 1982, Chopra et al., 1972, Brown et al., 1970, Ekins, 1998). In contrast to LC-MS/MS, early IA did not have multi-analyte capability. The ELISA-based (enzyme-linked immunosorbent assay) methods in principle use one antibody to immobilize the antigen and a further antibody as a detection antibody. The latter antibody is labelled with
Summary
Tandem MS has proved to be an extremely valuable bioanalytical tool for THM, and its combination with liquid chromatography has become the method of choice for many laboratories. Although the validation of published LC-MS/MS methods has become a routine requirement, there is no generally accepted “golden rule” as to how a validation procedure should be defined, and there are clearly areas of QA that can be improved. In particular, the use of MRM quantitation and confirmation ions and their
Funding
This work is supported by grants from the Deutsche Forschungsgemeinschaft within the DFG-SPP 1629 ThyroidTransAct KO 922/17-1/2 to JK.
Conflict of interest statement
The authors declare that there are no competing interests.
Acknowledgements
We thank Mr. Ray Monk for expert technical assistance.
References (72)
- et al.
Validation of a liquid chromatography-tandem mass spectrometry method to enable quantification of 3-iodothyronamine from serum
J. Chromatogr. A
(2008) - et al.
Stir bar sorptive extraction combined with high performance liquid chromatography-ultraviolet/inductively coupled plasma mass spectrometry for analysis of thyroxine in urine samples
J. Chromatogr. A
(2013) - et al.
Simultaneous quantification of free triiodothyronine and free thyroxine by isotope dilution tandem mass spectrometry
Clin. Biochem.
(2007) - et al.
Identification and quantification of 3-iodothyronamine metabolites in mouse serum using liquid chromatography-tandem mass spectrometry
J. Chromatogr. A
(2012) - et al.
Stable isotope-dilution liquid chromatography/tandem mass spectrometry method for determination of thyroxine in saliva
J. Chromatogr. B Anal. Technol. Biomed. Life Sci.
(2011) - et al.
3,3'-Diiodothyronine concentrations in hospitalized or thyroidectomized patients: results from a pilot study
Endocr. Pract.
(2014) - et al.
Validated ultra high performance liquid chromatography-tandem mass spectrometry method for quantitative analysis of total and free thyroid hormones in bovine serum
J. Chromatogr. A
(2014) - et al.
A method for the analysis of six thyroid hormones in thyroid gland by liquid chromatography-tandem mass spectrometry
J. Chromatogr. B Anal. Technol. Biomed. Life Sci.
(2010) - et al.
Development of Standard Reference Materials to support assessment of iodine status for nutritional and public health purposes
Am. J. Clin. Nutr.
(2016) - et al.
ApoB-100-containing lipoproteins are major carriers of 3-iodothyronamine in circulation
J. Biol. Chem.
(2012)
Thyroid hormone testing by tandem mass spectrometry
Clin. Biochem.
The measurement of free thyroxine by isotope dilution tandem mass spectrometry, Clinica chimica acta
Int. J. Clin. Chem.
Isotope dilution tandem mass spectrometric method for T4/T3, Clinica chimica acta
Int. J. Clin. Chem.
Determination of free thyroid hormones
Best. Pract. Res. Clin. Endocrinol. Metab.
The expanding role of tandem mass spectrometry in optimizing diagnosis and treatment of thyroid disease
Adv. Clin. Chem.
A “Step-Up” approach for harmonization, Clinica chimica acta
Int. J. Clin. Chem.
Radioimmunoassay of thyroxine (T4), 3,5,3'-triiodothyronine (T3), 3,3',5'-triiodothyronine (reverse T3, rT3), and 3,3'-diiodothyronine (T2)
Methods Enzymol.
An online solid-phase extraction-liquid chromatography-tandem mass spectrometry method to study the presence of thyronamines in plasma and tissue and their putative conversion from 13C6-thyroxine
J. Endocrinol.
Determination of thyroid hormones and their metabolites in tissue using SPE UPLC-tandem MS
Biomed. Chromatogr.
Triple bioaffinity mass spectrometry concept for thyroid transporter ligands
Anal. Chem.
Laboratory medicine practice guidelines. Laboratory support for the diagnosis and monitoring of thyroid disease
Thyroid
Guidance for industry: bioanalytical method validation
Liquid chromatography/mass spectrometry: the method of choice for the qualitative and quantitative analysis of environmental pollutants
3-Iodothyronamine: a novel hormone controlling the balance between glucose and lipid utilisation
J. Comp. Physiol. B
A radioimmunoassay for serum tri-iodothyronine
J. Endocrinol.
Measurement of serum T4 concentration by high performance liquid chromatography
J. Clin. Endocrinol. Metab.
Molecular aspects of thyroid hormone actions
Endocr. Rev.
An improved radioimmunoassay of triiodothyronine in serum: its application to clinical and physiological studies
J. Lab. Clin. Med.
Isotope dilution-liquid chromatography/electrospray ionization-tandem mass spectrometry for the determination of serum thyroxine as a potential reference method
Rapid Commun. mass Spectrom. RCM
Ligand assays: from electrophoresis to miniaturized microarrays
Clin. Chem.
Progress in standardizing and harmonizing thyroid function tests
Am. J. Clin. Nutr.
Detection of 3-iodothyronamine in human patients: a preliminary study
J. Clin. Endocrinol. Metab.
A new linear ion trap mass spectrometer
Rapid Commun. Mass Sp.
Quantification of 11 thyroid hormones and associated metabolites in blood using isotope-dilution liquid chromatography tandem mass spectrometry
Anal. Bioanal. Chem.
High-pressure liquid-chromatography of amino-acids, peptides and proteins .14. High-pressure liquid-chromatography of thyromimetic iodoamino acids
J. Liq. Chromatogr.
Thyronamines and derivatives: physiological relevance, pharmacological actions, and future research directions
Thyroid
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The 3-iodothyronamine (T1AM) and the 3-iodothyroacetic acid (TA1) indicate a novel connection with the histamine system for neuroprotection
2021, European Journal of PharmacologyCitation Excerpt :These uncertainness mostly derive from the methodological issues encountered in the assessment of T1AM plasma levels and from the incomplete knowledge of the step-to step enzyme pattern involved in T1AM biosynthesis. In respect of this latter, an extensive critical revision of the different protocols for detecting circulating T1AM levels is available for anyone interested (Richards et al., 2017) whereas a novel analytical method of detection has been published while this review was writing (Li et al., 2021). In the meanwhile, the pharmacological profile of the amine delineated progressively indicating from one side the amine maintains some of the effects of a trace amine, as the capacity to modulate the aminergic transmission (Lam et al., 2015), from the other it is proposing as a hormone with metabolic and behavioral profiles.
Rapid determination of thyroid hormones in blood plasma from Glaucous gulls and Baikal seals by HybridSPE®-LC-MS/MS
2021, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life SciencesCitation Excerpt :In particular, the transformation of T4 to T3 has a strong impact on T3 concentrations, which are significantly (even 2 orders of magnitude) lower than those of T4 in the blood of vertebrates [19]. Moreover, another major challenge in LC-MS/MS bioanalysis of THs is the uncertainty of quantification due to matrix effects that derive mainly from the high endogenous protein and phospholipid content of serum and plasma in contrast to the natural occurring trace concentrations of THs [13,16]. In this study, a rapid methodology that used the hybrid solid phase extraction (HybridSPE®) technique tailored to LC–electrospray (ESI+)–MS/MS was developed for the analysis of the total concentrations of four THs, T4, T3, rT3 and T2.
A new method for measuring thyroid hormones using nano-LC-MS/MS
2018, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life SciencesCitation Excerpt :Therefore methods with very low detection limits are needed. Richards et al. [7] recently summarized studies measuring plasma THs by LC-MS/MS (mostly in humans and rat/mouse) and associated pitfalls. For example, Hansen et al. [8] developed the simultaneous measurement of 11 TH metabolites in frog plasma, and Noyes et al. [9] validated the method in teleost fish.
Determination of thyroid hormones in placenta using isotope-dilution liquid chromatography quadrupole time-of-flight mass spectrometry
2018, Journal of Chromatography ACitation Excerpt :This method is of high sensitivity, but lack selectivity due to nonspecific interferences [18–20]. Methods based on tandem mass spectrometry offer a better specificity and accuracy [21], and have been used in various samples such as human and animal serum/plasma and tissues [20,22–29]. Although it is of significant importance, only two methods were reported for TH analysis in human placenta using IA method or liquid chromatography with tandem mass spectrometry technique (LC–MS/MS) [30,31].
T-Time again – The diversity of actions of thyroid hormones (TH), their aminergic (TAM) and acetic acid (TAc) metabolites
2017, Molecular and Cellular EndocrinologyMass spectrometry in the diagnosis of thyroid disease and in the study of thyroid hormone metabolism
2022, Mass Spectrometry Reviews