Measurement of free thyroid hormones in serum by column adsorption chromatography and radioimmunoassay

Journal of Endocrinological Investigation
P B RomelliL Vancheri

Abstract

A new method for the assay of free thyroid hormones in human serum is described. This method is based on a chromatographic adsorption process of thyroid hormones onto a Sephadex LH-20 resin column. Protein fractions are eliminated by washing columns, adsorbed hormones are eluted with methanol and determined by radioimmunoassay. It was demonstrated that under the experimental conditions adopted the presence of the resin R does not significantly change the free hormone level in the serum, and the amount of hormone adsorbed onto the resin HR is exclusively in function of the free hormone concentration [H], according to a linear relationship: HR = phi [H], where phi is the resin adsorption constant K ads multiplied by the number of resin binding sites nR. The phi value, experimentally determined, was 32 ml for T3 and 58 ml for T4, when 150 mg resin were used. The method sensitivity was 0.3 pg/ml for FT3 and 0.6 pg/ml for FT4. The within-assay reproducibility was about 5% (CV) and the between-assay reproducibility was about 6% (CV), both for FT3 and FT4. FT3 and FT4 levels, in 96 normal subjects, were 3.9 +/- 0.7 pg/ml (mean +/- SD) and 11.1 +/- 1.9 pg/ml (mean +/- SD) respectively.

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Citations

Oct 1, 1979·Clinical Endocrinology·M LangerC Costanza
May 1, 1992·Journal of Endocrinological Investigation·A ParacchiP Raineri
Aug 1, 1986·Journal of Endocrinological Investigation·A PacchiarottiA Pinchera
Jun 1, 1983·Journal of Endocrinological Investigation·M LangerG Madeddu
Apr 1, 1986·Journal of Endocrinological Investigation·A PacchiarottiA Pinchera
Nov 1, 1988·Journal of Endocrinological Investigation·F MarongiuE Martino
Dec 1, 1987·Journal of Endocrinological Investigation·S BalzanoE Martino
Oct 1, 1984·Clinical Endocrinology·G MadedduM Langer
Jul 1, 1993·Journal of the American Geriatrics Society·I SzabolcsJ Herrmann
Feb 1, 1992·Journal of Endocrinological Investigation·A CrinòP Beck-Peccoz
Nov 1, 1991·Journal of Endocrinological Investigation·E MartinoL E Braverman
Dec 1, 1991·Journal of Endocrinological Investigation·L ChiovatoG Ferretti
Feb 1, 1983·Journal of Endocrinological Investigation·C FerrariS Frezzati
Nov 1, 1993·Journal of Endocrinological Investigation·F Aghini-LombardiE Martino
Jan 1, 1997·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·C MarcocciA Pinchera
Aug 17, 2006·Thyroid : Official Journal of the American Thyroid Association·Eugenio ReschiniPaolo Gerundini
Jan 1, 1984·European Journal of Clinical Pharmacology·E RivaR Urso
Sep 1, 1982·Annals of Clinical Biochemistry·J W Tuttlebee
Mar 1, 1982·Annals of Clinical Biochemistry·H A RossJ H Thijssen

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