Studies on Steroids in Urine of the Male Newborn

Abstract

Four different methods of isolation and purification were utilized to study steroids in urine of male newborns which was collected during the first 5 days of life. These methods included celite column, ion exchange column and thin-layer chromatography, solvolysis and enzyme hydrolysis with β-glucuronidase and aryl sulfatase. Procedural losses were evaluated by using radioactive internal standards. Final quantitation of each steroid was achieved by comparison of its chromatographic and quantitative behavior with the respective standard steroids on various gas-liquid chromatography systems, either as parent compound or as trimethylsilyl ether derivative. The following steroids were found in the amounts indicated: progesterone, 2.1 μg/l (pool I), 4.6 μg/l (pool III); pregnanediol, 625.0 μg/l (pool IIa), 605.0 μg/l (pool IIb glucuronide), 25.4 μg/l (pool IIb sulfate), 4.2 μg/l (pool IIb free), 729.0 μg/l (pool III); 16α-hydroxyprogesterone, 713.0 μg/l (pool III); 16α-hydroxypregnenolone, 14,000.0 μg/l (pool III); 16α-hydroxydehydro-epiandrosterone, 2,350.0 μg/l (pool III); 16-dehydroprogesterone, 155.0 μg/l (pool I), 21.2 μg/l (pool IIb glucuronide), 97.5 μg/l (pool IIb sulfate), 5.3 μg/l (pool III); 16-de-hydropregnenolone, 382.0 μg/l (pool I), 1,380 μg/l (pool IIb glucuronide), 172.0 μg/l (pool IIb sulfate), 174.0 μg/l (pool III); 16-dehydropregnanolone, 8.3 μg/l (pool I), 239.0 μg/l (pool IIb sulfate). Pregnenolone, pregnanolone, 17α-hydroxyprogesterone and 17α-hydroxypregnenolone could not be detected. The results support the concept that the steroid patterns of urine of the newborn and amniotic fluid are very similar and that the amniotic fluid steroid content is mainly dependent on fetal urinary steroid excretion. The data on Δ¹⁶-C₂₁ -steroids are discussed.

Development of an Enzyme Immunoassay for Serum 16-Dehydropregnenolone.

“…16-DHP was initially identified in the mono-sulfate conjugate fraction of neonatal feces by Gustafsson et al 3) Subsequently, this steroid has been identified in human placenta, 4) urine, [5][6][7][8] gall-bladder bile, 9) meconium 10) and amniotic fluid. 11) Previously we also identified 16-DHP in the serum of immature infants and we suggested that it is a form of the sulfate conjugate, 16-DHP sulfate (16DHPS).…”