Automated chromatographic system for the simultaneous measurement of plasma pregnenolone and 17-hydroxypregnenolone by radioimmunoassay.
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.
Riepe
F G
FG
Department of Paediatrics, Division of Paediatric Endocrinology, Christian-Albrechts University Kiel, Germany.
Wonka
S
S
Partsch
C J
CJ
Sippell
W G
WG
D006907:17-alpha-Hydroxypregnenolone / Q000097:blood*; D000328:Adult; D001331:Automation; D002648:Child; D002852:Chromatography, Ion Exchange / Q000295:instrumentation*; D005260:Female; D006801:Humans; D008297:Male; D011284:Pregnenolone / Q000097:blood*; D011863:Radioimmunoassay / Q000379:methods*; D012016:Reference Values; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D006907:17-alpha-Hydroxypregnenolone; D011284:Pregnenolone