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Correlation between Serum Levels of 3,3',5'-Triiodothyronine and Thyroid Hormones Measured by Liquid Chromatography-Tandem Mass Spectrometry and Immunoassay.

Sakai H, Nagao H, Sakurai M, Okumura T, Nagai Y, Shikuma J, Ito R, Imazu T, Miwa T, Odawara M - PLoS ONE (2015)

Bottom Line: To the best of our knowledge, there are no studies to date comparing results of the quantification of human serum rT3 between LC-MS/MS and RIA.Matrix effects were not observed with our method.Our LC-MS/MS system requires no manual cleanup operation, and the process after application of a sample is fully automated; furthermore, it was found to be highly sensitive, and superior in both precision and accuracy.

View Article: PubMed Central - PubMed

Affiliation: Department of Diabetes, Endocrinology and Metabolism, Tokyo Medical University, Tokyo, Japan.

ABSTRACT

Objective: For measuring serum 3,3',5'-triiodothyronine (rT3) levels, radioimmunoassay (RIA) has traditionally been used owing to the lack of other reliable methods; however, it has recently become difficult to perform. Meanwhile, liquid chromatography-tandem mass spectrometry (LC-MS/MS) has recently been attracting attention as a novel alternative method in clinical chemistry. To the best of our knowledge, there are no studies to date comparing results of the quantification of human serum rT3 between LC-MS/MS and RIA. We therefore examined the feasibility of LC-MS/MS as a novel alternative method for measuring serum rT3, thyroxine (T4), and 3,5,3'-triiodothyronine (T3) levels.

Methods: Assay validation was performed by LC-MS/MS using quality control samples of rT3, T4, and T3 at 4 various concentrations which were prepared from reference compounds. Serum samples of 50 outpatients in our department were quantified both by LC-MS/MS and conventional immunoassay for rT3, T4, and T3. Correlation coefficients between the 2 measurement methods were statistically analyzed respectively.

Results: Matrix effects were not observed with our method. Intra-day and inter-day precisions were less than 10.8% and 9.6% for each analyte at each quality control level, respectively. Intra-day and inter-day accuracies were between 96.2% and 110%, and between 98.3% and 108.6%, respectively. The lower limit of quantification was 0.05 ng/mL. Strong correlations were observed between the 2 measurement methods (correlation coefficient, T4: 0.976, p < 0.001; T3: 0.912, p < 0.001; rT3: 0.928, p < 0.001).

Conclusions: Our LC-MS/MS system requires no manual cleanup operation, and the process after application of a sample is fully automated; furthermore, it was found to be highly sensitive, and superior in both precision and accuracy. The correlation between the 2 methods over a wide range of concentrations was strong. LC-MS/MS is therefore expected to become a useful tool for clinical diagnosis and research.

No MeSH data available.


Chemical structures of [2H5]T4 (A), T4 (B), T3 (C), and rT3 (D).
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pone.0138864.g001: Chemical structures of [2H5]T4 (A), T4 (B), T3 (C), and rT3 (D).

Mentions: L-Thyroxine-[L-tyrosine-2H5] (Fig 1A) was purchased from IsoSciences, LLC (King of Prussia, PA). T4, T3, and rT3 (Fig 1B, 1C and 1D, respectively) were purchased from Sigma-Aldrich Co. (Piscataway, NJ). Thyroid hormone-free human serum was purchased from Golden West Biologicals (DDC Mass Spect GoldĀ®, Temecula, CA). All other commercially available chemicals and reagents were of the highest analytical grade.


Correlation between Serum Levels of 3,3',5'-Triiodothyronine and Thyroid Hormones Measured by Liquid Chromatography-Tandem Mass Spectrometry and Immunoassay.

Sakai H, Nagao H, Sakurai M, Okumura T, Nagai Y, Shikuma J, Ito R, Imazu T, Miwa T, Odawara M - PLoS ONE (2015)

Chemical structures of [2H5]T4 (A), T4 (B), T3 (C), and rT3 (D).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591014&req=5

pone.0138864.g001: Chemical structures of [2H5]T4 (A), T4 (B), T3 (C), and rT3 (D).
Mentions: L-Thyroxine-[L-tyrosine-2H5] (Fig 1A) was purchased from IsoSciences, LLC (King of Prussia, PA). T4, T3, and rT3 (Fig 1B, 1C and 1D, respectively) were purchased from Sigma-Aldrich Co. (Piscataway, NJ). Thyroid hormone-free human serum was purchased from Golden West Biologicals (DDC Mass Spect GoldĀ®, Temecula, CA). All other commercially available chemicals and reagents were of the highest analytical grade.

Bottom Line: To the best of our knowledge, there are no studies to date comparing results of the quantification of human serum rT3 between LC-MS/MS and RIA.Matrix effects were not observed with our method.Our LC-MS/MS system requires no manual cleanup operation, and the process after application of a sample is fully automated; furthermore, it was found to be highly sensitive, and superior in both precision and accuracy.

View Article: PubMed Central - PubMed

Affiliation: Department of Diabetes, Endocrinology and Metabolism, Tokyo Medical University, Tokyo, Japan.

ABSTRACT

Objective: For measuring serum 3,3',5'-triiodothyronine (rT3) levels, radioimmunoassay (RIA) has traditionally been used owing to the lack of other reliable methods; however, it has recently become difficult to perform. Meanwhile, liquid chromatography-tandem mass spectrometry (LC-MS/MS) has recently been attracting attention as a novel alternative method in clinical chemistry. To the best of our knowledge, there are no studies to date comparing results of the quantification of human serum rT3 between LC-MS/MS and RIA. We therefore examined the feasibility of LC-MS/MS as a novel alternative method for measuring serum rT3, thyroxine (T4), and 3,5,3'-triiodothyronine (T3) levels.

Methods: Assay validation was performed by LC-MS/MS using quality control samples of rT3, T4, and T3 at 4 various concentrations which were prepared from reference compounds. Serum samples of 50 outpatients in our department were quantified both by LC-MS/MS and conventional immunoassay for rT3, T4, and T3. Correlation coefficients between the 2 measurement methods were statistically analyzed respectively.

Results: Matrix effects were not observed with our method. Intra-day and inter-day precisions were less than 10.8% and 9.6% for each analyte at each quality control level, respectively. Intra-day and inter-day accuracies were between 96.2% and 110%, and between 98.3% and 108.6%, respectively. The lower limit of quantification was 0.05 ng/mL. Strong correlations were observed between the 2 measurement methods (correlation coefficient, T4: 0.976, p < 0.001; T3: 0.912, p < 0.001; rT3: 0.928, p < 0.001).

Conclusions: Our LC-MS/MS system requires no manual cleanup operation, and the process after application of a sample is fully automated; furthermore, it was found to be highly sensitive, and superior in both precision and accuracy. The correlation between the 2 methods over a wide range of concentrations was strong. LC-MS/MS is therefore expected to become a useful tool for clinical diagnosis and research.

No MeSH data available.