Automated chromatographic system for the simultaneous measurement of plasma pregnenolone and 17-hydroxypregnenolone by radioimmunoassay

Journal of Chromatography B: Biomedical Sciences and Applications Pub Date : 2001-11-05 DOI:10.1016/S0378-4347(01)00373-5

F.G Riepe, S Wonka, C.-J Partsch, W.G Sippell

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引用次数: 15

Abstract

A new, simple, rapid and highly practicable automated chromatographic system for the separation, and a sensitive radioimmunoassay system for the subsequent measurement of pregnenolone and 17-hydroxypregnenolone has been developed. Pregnenolone and 17-hydroxypregnenolone were extracted with methylene chloride and separated from cross-reacting steroids by mechanised Sephadex-LH20 multi-column chromatography. Anti-pregnenolone and anti-17-hydroxypregnenolone were obtained by immunising rabbits with pregnenolone-20-oxime-BSA and 17-hydroxypregnenolone-20-oxime-BSA. The lower detection limit of the assay is 0.15 and 0.28 nmol/l for pregnenolone and 17-hydroxypregnenolone, respectively. Normal values for this assay in young male adults, in adult females, and in prepubertal boys and girls were established as a basis for the functional diagnosis of androgen excess syndromes/steroidogenesis defects.

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用放射免疫法同时测定血浆孕烯醇酮和17-羟基孕烯醇酮的自动色谱系统

建立了一种新的、简单、快速和高度实用的自动色谱分离系统，以及一种灵敏的放射免疫分析系统，用于孕烯醇酮和17-羟基孕烯醇酮的后续测量。用二氯甲烷提取孕烯醇酮和17-羟基孕烯醇酮，用机械Sephadex-LH20多柱层析从交叉反应类固醇中分离。用孕烯醇酮-20-肟- bsa和17-羟基孕烯醇酮-20-肟- bsa免疫家兔获得抗孕烯醇酮和抗17-羟基孕烯醇酮。孕烯醇酮和17-羟基孕烯醇酮的检测下限分别为0.15和0.28 nmol/l。该检测在年轻男性、成年女性和青春期前男孩和女孩中的正常值被确立为雄激素过量综合征/类固醇生成缺陷功能诊断的基础。

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Journal of Chromatography B: Biomedical Sciences and Applications

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