Abstract
Urine was chromatographed on DEAE-Sephadex. The unconjugated steroids and the steroids liberated by β-glucuronidase hydrolysis from the glucuronide fraction were isolated by Amberlite XAD-2 chromatography. After TLC the polar steroids (RF ≤ THE) were subjected to HPLC. The different fractions obtained by HPLC were analysed by GC-MS. 6α-hydroxy-tetrahydrocortisone (6α-OHTHE) and the 6α-hydroxy-cortolones (6α-OH-20αC and 6α-OH-20βC) were identified in the neonatal urine by comparing these compounds with standard steroids previously synthesised from 6α-hydroxy-cortisone. Steroids were quantitated by GC or mass fragmentography. Internal standards were used to correct for losses during the entire procedure. The average amounts (μg/kg body weight/24 hr) excreted on the second day of life by six fullterm infants were:
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