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Clinical Chemistry 25: 1757-1760, 1979;
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Clinical Chemistry, Vol 25, 1757-1760, Copyright © 1979 by American Association for Clinical Chemistry

Rapid analysis for iodotyrosines and iodothyronines in thyroglobulin by reversed-phase liquid chromatography

NM Alexander and M Nishimoto

We describe a 10-min reversed-phase "high-pressure' liquid- chromatographic procedure for measuring tyrosine, monoiodotyrosine, diiodotyrosine, 3,5-diiodothyronine, 3,5,3'-triiodothyronine, 3,3',5'- triiodothyronine, and thyroxine. Resolution and quantitation of a mixture of these amino acids were excellent on LiChrosorb (Altex) RP-8 with isocratic elution (1.5 mL/min) with acetonitrile/water/glacial acetic acid (50/49/1 by vol). As little as 100 ng of each iodoamino acid could be detected and quantitated with a conventional 1-cm, flow- through spectrophotometric (254-nm) detector coupled to a 10-mV strip- chart recorder. Analyses for monoiodotyrosine, diiodotyrosine, 3,5,3'- triiodothyronine, and thyroxine in hog and beef thyroglobulin hydrolysates (sequential digestion with pronase and aminopeptidase) agreed well with results by anion-exchange chromatography and by competitive radioassays. To prevent interference by tryptophan in the analysis for diiodotyrosine, we batch-separated the iodoamino acids by anion-exchange chromatography before the procedure. The procedure we describe seems generally useful for detection and quantitation of thyroid hormones, thyroid hormone metabolites, and iodotyrosines.




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Copyright © 1979 by the American Association for Clinical Chemistry.