Romelli P B, Pennisi F, Vancheri L
J Endocrinol Invest. 1979 Jan-Mar;2(1):25-40. doi: 10.1007/BF03349272.
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.
本文描述了一种检测人血清中游离甲状腺激素的新方法。该方法基于甲状腺激素在Sephadex LH - 20树脂柱上的色谱吸附过程。通过洗涤柱子去除蛋白质组分，用甲醇洗脱吸附的激素，并用放射免疫分析法进行测定。结果表明，在所采用的实验条件下，树脂R的存在不会显著改变血清中的游离激素水平，并且根据线性关系：HR = phi [H]，吸附在树脂HR上的激素量仅取决于游离激素浓度[H]，其中phi是树脂吸附常数K ads乘以树脂结合位点数量nR。当使用150 mg树脂时，实验测定的phi值对于T3为32 ml，对于T4为58 ml。该方法对FT3的灵敏度为0.3 pg/ml，对FT4的灵敏度为0.6 pg/ml。FT3和FT4的批内重复性约为5%（CV），批间重复性约为6%（CV）。在96名正常受试者中，FT3和FT4水平分别为3.9 +/- 0.7 pg/ml（平均值 +/- 标准差）和11.1 +/- 1.9 pg/ml（平均值 +/- 标准差）。
