Analysis of thyroid hormones in serum by liquid chromatography-tandem mass spectrometry

Thyroid hormones are essential hormones for regulating growth and development in humans and wildlife. Methods to monitor precise and low levels of these hormones in serum and tissues are needed to assess overall health, whether from disease considerations or possibly from environmental contaminant exposures. Common and routine methods typically rely upon radioimmunoassays, which can be expensive, and typically only measure thyroxine and 3,3′,5-triidothyronine, which can be a limitation in fully evaluating impacts on thyroid regulation. In this study we developed a liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for the simultaneous analysis of five thyroid hormones—thyroxine, 3,3′,5-triidothyronine, 3,3′,5′-triiodothyronine, 3,3′-diiodothyronine, and 3,5-diiodothyronine—in serum samples. The LC-MS/MS parameters were optimized and calibrated over a wide concentration range (1.0–500 ng/mL) with on-column detection limits of 1.5–7.0 pg. With use of spiked bovine serum samples, the mean method recoveries were calculated to be 81.3–111.9% with relative standard deviations of 1.2–9.6% at spiking levels ranging from 10 to 100 ng/mL. This method was compared with measurements made by standard radioimmunoassays and with measurements made in a serum Standard Reference Material (SRM 1951b). Development of this method expands the capacity to measure thyroid hormones by including a larger suite of thyroid hormones, and has promising applications for measuring catabolism of thyroid hormones in vitro.