Rapid, high-throughput method for the quantification of thyroid hormones in rat blood serum using isotope-dilution LC-MS/MS

Aim: Numerous guideline studies required for regulatory toxicity testing now include the measurement of the thyroid hormones 3,3 ',5-triiodo-L-thyronine (T3) and L-thyroxine (T4) in blood serum from rodents. A rapid, high-throughput method for the determination of the thyroid hormones T4 and T3 is reported. Materials & methods: Sample preparation is done using a 96-well microtiter plate format. Stable isotope analogs of both hormones are used as internal standards for study and quality control samples. Results & conclusion: The validated quantification levels are T3: 10 pg/ml and T4: 1 ng/ml, with CVs of <10% at the limit of quantification and up to 50*limit of quantification. The use of isotope analog internal standards eliminates the need for quantitative transfers and complete evaporations.