Rudolph M, Sakurada T, Fang S L, Vagenakis A G, Braverman L E, Ingbar S H
J Clin Endocrinol Metab. 1978 Jun;46(6):923-8. doi: 10.1210/jcem-46-6-923.
Chromatography of serum on columns of Sephadex G-25 superfine after the iv administration of 125I-labeled T4 consistently yielded labeled iodide, iodoprotein, T3, and a labeled peak that eluted from the column before the T3, termed "pre-T3." Much larger quantities of pre-T3 were generated after the iv administration of 125I-labeled T3 and rT3. The ratio of [125I]pre T3:[125I]T3 and [125I]pre T3:[125I]rT3 plateaued at approximately 10 h and 2 h, respectively, after the iv administration of the labeled hormone, and averaged approximately 15% in euthyroid subjects. As pre-T3 behaves like its precursors in the TCA precipitation-ethanol extraction or anion exchange chromatographic procedures used to separate labeled iodide and iodoprotein from administered labeled T3 and rT3, concentrations of labeled hormone measured by these techniques will be in error, because pre-T3 will be included. Thus, the MCR of labeled T3 and rT3 measured by Sephadex chromatography will always be higher than values obtained by the other two separative techniques and the magnitude of change will be similar to the ratio of pre-T3 to precursor T3 or rT3. The Sephadex chromatographic technique is laborious, but appears to be the method of choice where greatest accuracy of measurement is required. As the generation of pre-T3 from labeled T4 is sufficiently slow, the present chromatographic technique is not necessary in studying peripheral T4 turnover.
静脉注射125I标记的T4后,在Sephadex G - 25超细柱上对血清进行色谱分析,始终会产生标记碘化物、碘蛋白、T3以及一个在T3之前从柱上洗脱的标记峰,称为“前T3”。静脉注射125I标记的T3和反T3(rT3)后,会产生大量更多的前T3。静脉注射标记激素后,[125I]前T3:[125I]T3和[125I]前T3:[125I]rT3的比值分别在约10小时和2小时达到平台期,在甲状腺功能正常的受试者中平均约为15%。由于前T3在用于从注射的标记T3和rT3中分离标记碘化物和碘蛋白的三氯乙酸沉淀 - 乙醇萃取或阴离子交换色谱程序中表现得像其前体,因此用这些技术测量的标记激素浓度会有误,因为会包含前T3。因此,通过Sephadex色谱法测量的标记T3和rT3的代谢清除率(MCR)将始终高于通过其他两种分离技术获得的值,且变化幅度将与前T3与前体T3或rT3的比值相似。Sephadex色谱技术费力,但在需要最高测量精度的情况下似乎是首选方法。由于从标记T4生成前T3足够缓慢,在研究外周T4周转时,目前的色谱技术并非必要。