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Unconjugated steroids were extracted with dichloromethane from the serum of pre-term neonates. Conjugated steroid fraction in the residue was separated and purified by high performance liquid chromatography. The purified steroid fraction was subjected to enzyme (arylsulfatase from Helix pomatia) or acid hydrolysis. Liberated steroid was analyzed by gas chromatography-mass spectrometry (GC/MS) and gas chromatography-Fourier transformation infrared spectroscopy (GC/FT-IR). All of the results obtained from GC/MS and GC/FT-IR analysis were identical with those of authentic 16-dehydropregnenolone (3 beta-hydroxy-5,16-pregnadien-20-one, 16-DHP). This is the first report to demonstrate the presence of 16-DHP in human blood.

Studies on Steroids in Fetuses and Neonates: Identification of 16-Dehydropregnenolone in the Circulation of Pre-term Neonates.