Pirke K M
Steroids. 1977 Jul;30(1):53-60. doi: 10.1016/0039-128x(77)90136-2.
A reliable radioimmunoassay for the determination of 5-androstene-3beta, 17beta-diol in plasma is described. Antisera were obtained by immunization of rabbits with 3beta, 17beta-dihydroxy-5-androsten-16-one coupled to bovine serum albumin in position 16. The antiserum was characterized by titer, affinity, and specificity. Only dehydroepiandrosterone (24.3 percent) and pregnenolone (2.7 percent) showed a small cross-reactivity. The assay method consisted of extraction with ether, thin-layer chromatography and endpoint determination. The reliability of the method was studied. The interassay variability was 7.5 percent at a concentration of 1.22 microgram/l. The limit of detection was 0.068 microgram/l. Specificity was achieved by chromatographic separation of the crossreacting steroids. Mass recovery experiments with 250 and 500 pg were performed, in which 99.0 + or - 4.6 percent of the smaller and 97.6 +/- 11.3 percent of the greater mass were recovered. In 45 healthy adult males plasma concentrations between 0.44 and 1.80 microgram/l were found. The median was 1.06 microgram/l. Stimulation of the Leydig cells with human chorionic gonadotropin (HCG) increased plasma concentrations by 93 percent (average in 12 males). Thereapeutic castration in 8 men caused an average decrease of 55.4 percent in plasma values.
本文描述了一种用于测定血浆中5-雄烯-3β,17β-二醇的可靠放射免疫分析法。通过用16位偶联牛血清白蛋白的3β,17β-二羟基-5-雄烯-16-酮免疫兔子获得抗血清。该抗血清通过效价、亲和力和特异性进行表征。只有脱氢表雄酮（24.3%）和孕烯醇酮（2.7%）显示出较小的交叉反应性。测定方法包括用乙醚萃取、薄层色谱法和终点测定。研究了该方法的可靠性。在浓度为1.22微克/升时，批间变异为7.5%。检测限为0.068微克/升。通过对交叉反应类固醇进行色谱分离实现了特异性。进行了250和500皮克的质量回收实验，其中较小质量的回收率为99.0±4.6%，较大质量的回收率为97.6±11.3%。在45名健康成年男性中，血浆浓度在0.44至1.80微克/升之间。中位数为1.06微克/升。用人绒毛膜促性腺激素（HCG）刺激睾丸间质细胞可使血浆浓度升高93%（12名男性的平均值）。8名男性进行治疗性去势后，血浆值平均下降55.4%。
