de Man A J, Benraad T J
Clin Chim Acta. 1977 Sep 1;79(2):489-501. doi: 10.1016/0009-8981(77)90446-6.
A simple and reliable radioimmunoassay for the measurement of the aldosterone secretion rate has been developed. In this method the urinary pH 1-hydrolyzed aldosterone is acetylated to its 21-monoacetate and purified by a single paper chromatogram. The isolated aldosterone 21-monoacetate is radioimmunoassayed using the N.I.H. antiserum lot 088. From the value obtained and the tritium amount present in the eluate the specific activity is calculated. The following results disclosed the reliability of the method: 1. The values obtained were in perfect agreement with values obtained by a double isotope assay. 2. The aldosterone secretion rate values calculated did not change upon extension of chromatographic steps. 3. Immunochromatographic analysis showed a sharp peak in the aldosterone 21-monoacetate area without significant background immunoreactivity. 4. The steroid concentration measured decreased linearly with dilution of the paper eluate. 5. Identical values were obtained with three antisera, raised against different aldosterone immunogens. In our laboratory no reliable results could be obtained when the conversion to the aldosterone 21-monoacetate was omitted and the specific activity was calculated after radioimmunoassay of chromatographically purified, pH 1-hydrolyzed, aldosterone per se. In that case the values obtained were systematically lower than the double isotope values, a problem which is discussed.
已开发出一种用于测量醛固酮分泌率的简单可靠的放射免疫测定法。在该方法中，尿液中pH 1水解的醛固酮被乙酰化为其21 - 单乙酸酯，并通过单次纸色谱法进行纯化。使用美国国立卫生研究院（N.I.H.）批号为088的抗血清对分离出的醛固酮21 - 单乙酸酯进行放射免疫测定。根据所得值和洗脱液中存在的氚量计算比活性。以下结果揭示了该方法的可靠性：1. 所得值与通过双同位素测定法获得的值完全一致。2. 计算出的醛固酮分泌率值在色谱步骤延长时没有变化。3. 免疫色谱分析显示醛固酮21 - 单乙酸酯区域有一个尖锐峰，且背景免疫反应不明显。4. 测量的类固醇浓度随纸洗脱液的稀释呈线性下降。5. 用针对不同醛固酮免疫原产生的三种抗血清获得了相同的值。在我们实验室中，如果省略向醛固酮21 - 单乙酸酯的转化，并且在对色谱纯化的、pH 1水解的醛固酮本身进行放射免疫测定后计算比活性，则无法获得可靠结果。在那种情况下，所得值系统性地低于双同位素值，对此问题进行了讨论。
