Abstract
A relatively simple new technique is reported for the separation and measurement of the following Δ 5 steroids in human fetal blood, amniotic fluid and infant and adult urine: 3β-hydroxypregn-5-en-20-one, 3β-hydroxyandrost-5-en-17-one, 3β,21,-dihydroxypregn-5-en-20-one, 3β,17α(and 17β)-dihydroxyandrost-5-ene, 3β,17β-dihydroxyandrost-5-en-16-one, 3β, 16α-dihydroxyandrost-5-en-17-one, 3β,16α-dihydroxypregn-5-en-20-one, 3β,16α,17β-trihydroxy-androst-5-ene and two unidentified compounds. After enzyme hydrolysis and solvolysis the steroids are extracted from the biological material and separated on thin-layer silica-gel chromatograms. Assay is by direct photo-electric scanning of the coloured bands developed on the chromatograms by an antimony trichloride reagent. The identity of the compounds has been checked by gas chromatography-mass spectroscopy and further thin-layer chromatography. The accuracy, specificity and precision of the method have been evaluated. The total urinary excretion of the compounds when expressed in μg/24 hr/m 2 body area is approximately 10 times that of the adult. The 2 unknown compounds, 21-hydroxypregnenolone and 16-hydroxypregnenolone are undetectable in adult urine. In cases of the adrenogenital syndrome the excretion of 16-hydroxypregnenolone was markedly increased.
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