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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.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.LC-MS/MS is therefore expected to become a useful tool for clinical diagnosis and research.

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.


Comparison of the LC/MS/MS method with the RIA method for the measurement of serum rT3.y = 2.48x + 0.0456, r = 0.928, p < 0.001.
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pone.0138864.g003: Comparison of the LC/MS/MS method with the RIA method for the measurement of serum rT3.y = 2.48x + 0.0456, r = 0.928, p < 0.001.

Mentions: For serum T4 and T3 levels determined by LC-MS/MS and ECLIA, very strong correlations between the 2 measurement methods were obtained (r = 0.976, p < 0.001; r = 0.912, p < 0.001, respectively) (Fig 2A and 2B). Regarding the measurements of rT3 concentrations, 4 samples were found to be lower than 0.05 ng/mL (LLOQ) by LC-MS/MS, and 8 samples were higher than 0.72 ng/mL (maximum concentration of precision) by RIA. We therefore excluded those 12 samples, and statistically analyzed the remaining data (n = 38). A strong correlation was also observed between the measurements obtained by LC-MS/MS and by RIA (r = 0.928, p < 0.001) (Fig 3).


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)

Comparison of the LC/MS/MS method with the RIA method for the measurement of serum rT3.y = 2.48x + 0.0456, r = 0.928, p < 0.001.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0138864.g003: Comparison of the LC/MS/MS method with the RIA method for the measurement of serum rT3.y = 2.48x + 0.0456, r = 0.928, p < 0.001.
Mentions: For serum T4 and T3 levels determined by LC-MS/MS and ECLIA, very strong correlations between the 2 measurement methods were obtained (r = 0.976, p < 0.001; r = 0.912, p < 0.001, respectively) (Fig 2A and 2B). Regarding the measurements of rT3 concentrations, 4 samples were found to be lower than 0.05 ng/mL (LLOQ) by LC-MS/MS, and 8 samples were higher than 0.72 ng/mL (maximum concentration of precision) by RIA. We therefore excluded those 12 samples, and statistically analyzed the remaining data (n = 38). A strong correlation was also observed between the measurements obtained by LC-MS/MS and by RIA (r = 0.928, p < 0.001) (Fig 3).

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.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.LC-MS/MS is therefore expected to become a useful tool for clinical diagnosis and research.

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.