Skip to main content
Journal cover image

Rapid method for the measurement of circulating thyroid hormones in low volumes of teleost fish plasma by LC-ESI/MS/MS.

Publication ,  Journal Article
Noyes, PD; Lema, SC; Roberts, SC; Cooper, EM; Stapleton, HM
Published in: Analytical and bioanalytical chemistry
January 2014

Thyroid hormones are critical regulators of normal development and physiological functioning in all vertebrates. Radioimmunoassay (RIA) approaches have been the method of choice for measuring circulating levels of thyroid hormones in vertebrates. While sensitive, RIA-based approaches only allow for a single analyte measurement per assay, can lack concordance across platforms and laboratories, and can be prone to analytical interferences especially when used with fish plasma. Ongoing advances in liquid chromatography tandem mass spectrometry (LC/MS/MS) have led to substantial decreases in detection limits for thyroid hormones and other biomolecules in complex matrices, including human plasma. Despite these advances, current analytical approaches do not allow for the measurement of native thyroid hormone in teleost fish plasma by mass spectrometry and continue to rely on immunoassay. In this study, we developed a new method that allows for the rapid extraction and simultaneous measurement of total T4 (TT4) and total T3 (TT3) in low volumes (50 μL) of fish plasma by LC/MS/MS. Methods were optimized initially in plasma from rainbow trout (Oncorhynchus mykiss) and applied to plasma from other teleost fishes, including fathead minnows (Pimephales promelas), mummichogs (Fundulus heteroclitus), sockeye salmon (Oncorhynchus nerka), and coho salmon (Oncorhynchus kisutch). Validation of method performance with T4- and T3-spiked rainbow trout plasma at 2 and 4 ng/mL produced mean recoveries ranging from 82 to 95 % and 97 to 105 %, respectively. Recovery of (13)C12-T4 internal standard in plasma extractions was: 99 ± 1.8 % in rainbow trout, 85 ± 11 % in fathead minnow, 73 ± 5.0 % in mummichog, 73 ± 1.7 % in sockeye salmon, and 80 ± 8.4 % in coho salmon. While absolute levels of thyroid hormones measured in identical plasma samples by LC/MS/MS and RIA varied depending on the assay used, T4/T3 ratios were generally consistent across both techniques. Less variability was measured among samples subjected to LC/MS/MS suggesting a more precise estimate of thyroid hormone homeostasis in the species targeted. Overall, a sensitive and reproducible method was established that takes advantage of LC/MS/MS techniques to rapidly measure TT4 and TT3 with negligible interferences in low volumes of plasma across a variety of teleost fishes.

Duke Scholars

Published In

Analytical and bioanalytical chemistry

DOI

EISSN

1618-2650

ISSN

1618-2642

Publication Date

January 2014

Volume

406

Issue

3

Start / End Page

715 / 726

Related Subject Headings

  • Trout
  • Time Factors
  • Thyroid Hormones
  • Spectrometry, Mass, Electrospray Ionization
  • Molecular Structure
  • Limit of Detection
  • Fishes
  • Chromatography, Liquid
  • Blood Chemical Analysis
  • Animals
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Noyes, P. D., Lema, S. C., Roberts, S. C., Cooper, E. M., & Stapleton, H. M. (2014). Rapid method for the measurement of circulating thyroid hormones in low volumes of teleost fish plasma by LC-ESI/MS/MS. Analytical and Bioanalytical Chemistry, 406(3), 715–726. https://doi.org/10.1007/s00216-013-7528-3
Noyes, Pamela D., Sean C. Lema, Simon C. Roberts, Ellen M. Cooper, and Heather M. Stapleton. “Rapid method for the measurement of circulating thyroid hormones in low volumes of teleost fish plasma by LC-ESI/MS/MS.Analytical and Bioanalytical Chemistry 406, no. 3 (January 2014): 715–26. https://doi.org/10.1007/s00216-013-7528-3.
Noyes PD, Lema SC, Roberts SC, Cooper EM, Stapleton HM. Rapid method for the measurement of circulating thyroid hormones in low volumes of teleost fish plasma by LC-ESI/MS/MS. Analytical and bioanalytical chemistry. 2014 Jan;406(3):715–26.
Noyes, Pamela D., et al. “Rapid method for the measurement of circulating thyroid hormones in low volumes of teleost fish plasma by LC-ESI/MS/MS.Analytical and Bioanalytical Chemistry, vol. 406, no. 3, Jan. 2014, pp. 715–26. Epmc, doi:10.1007/s00216-013-7528-3.
Noyes PD, Lema SC, Roberts SC, Cooper EM, Stapleton HM. Rapid method for the measurement of circulating thyroid hormones in low volumes of teleost fish plasma by LC-ESI/MS/MS. Analytical and bioanalytical chemistry. 2014 Jan;406(3):715–726.
Journal cover image

Published In

Analytical and bioanalytical chemistry

DOI

EISSN

1618-2650

ISSN

1618-2642

Publication Date

January 2014

Volume

406

Issue

3

Start / End Page

715 / 726

Related Subject Headings

  • Trout
  • Time Factors
  • Thyroid Hormones
  • Spectrometry, Mass, Electrospray Ionization
  • Molecular Structure
  • Limit of Detection
  • Fishes
  • Chromatography, Liquid
  • Blood Chemical Analysis
  • Animals