Gas chromatographic analysis of steroids using Sp-2100 stationary phase revealed that all urine samples from 34 healthy children (aged 4-14 years) and from 35 adults contained 5-pregnene-3 beta, 20 alpha-diol, which was also identified by GC-MS. The extent of pregnenediol excretion continuously increased from prepuberty to adult age. The observed increase in pregnenediol excretion during prepuberty points to an enhanced biosynthetic activity of the adrenal gland in the years preceding sexual maturation.