A Method for the Separation and Estimation of Neutral Steroids in the Urine of Newborn Infants

Abstract

SUMMARY A method is described for the separation and estimation of neutral steroids in infant urine. The free steroids are obtained by a two-stage enzyme hydrolysis. Following preliminary chromatography, the extracts containing steroids of low and medium polarity are suitable for final resolution. The extract containing steroids of high polarity requires further purification by partition between ether-ethanol and aqueous ammonium sulphate, followed by reversed-phase paper chromatography. Final resolution of steroids in fractions of different polarity is achieved using Bush aqueous methanol systems. Losses incurred in the method are estimated by the use of 4-14C steroids.

3 : 21-Dihydroxypregn-5-ene-20-one in Urine of Normal Newborn Infants and in Third Day Urine of Child with Deficiency of 3 -hydroxysteroid-dehydrogenase

“…The first is that there may be increased utilization by the newborn infant of the biosynthetic pathway to corticosterone via 21hydroxypregnenolone, with resultant overspill of some of the intermediary steroid into the urine. This hypothesis is in agreement with the relatively higher output of corticosterone and its metabolites than of cortisol and its metabolites demonstrable in infancy urine (Birchall et al, 1961;Cathro et al, 1963;Exley and Norymberski, 1963). Supportive evidence for increased use of the pathway via 21-hydroxypregnenolone during disproportionate corticosterone secretion by the adrenal gland has been given in the present study by analysis of pregnancy urine.…”

“…in an affected infant at the age of 2 weeks but did not report the DHA excretion in a case in which the diagnosis was made at 7 days. Further information on the excretion of DHA during the early days of life in infants with this syndrome would be advantageous to the endocrinologist faced with making a decision on the optimum time for establishing the diagnosis of the disease by means of urinary steroid analysis, since, if a slight or relative deficiency of the enzyme is postulated in the healthy normal infant, some explanation must exist for the minimal excretion of DHA in the urine of endocrinologically normal infants during the first week of life (Birchall et al, 1961;Cathro et al, 1963). It is noteworthy that DHA has been reported to occur in large amounts in meconium (Francis, Shen, and Kinsella, 1960) and to attain higher levels in cord blood than in maternal blood (Migeon, Keller, and Holmstrom, 1955;Colas, Heinrichs, and Tatum, 1964), so that the small amounts found in the urine in the first week of life in the normal child can only be explained by a major excretion of DHA in the form of hydroxylated metabolites such as 16oc-hydroxy-DHA (Reynolds, 1964).…”

“…The unusual patterns of excretion of steroids of adrenal or placental origin occurring in the urine of normal newborn infants have been the subject of previous reports (Birchall, Cathro, Forsyth, and Mitchell, 1961;Cathro, Birchall, Mitchell, and Forsyth, 1963) and have been interpreted as providing evidence that the metabolism of adrenal steroids in early infancy is unique. The presence of atypical urinary neutral steroids is characteristic of the first…”

Corticosteroid Secretion in the Monotreme Tachyglossus Aculeatus

“…Visual comparison was preferred to fluorimetry on paper, because of greater sensitivity and convenience. Similar preference has been expressed by Birchall, Cathro, Forsyth & Mitchell (1963). The lower limit for detection was Table 1.…”

Adrenocortical response to stress in newborn infants.

“…The adult cortex therefore must supply all the needs of the infant for the period of adaptation to extrauterine life. However, the high excretion of J5-steroids in the urine during the first week of life (Cathro et al, 1963(Cathro et al, , 1965Mitchell, 1967) is evidence that there is a relative insufficiency of J5-3fl-ol-dehydrogenase in the definitive cortex at this period, which does not exist in later life. On theoretical grounds, therefore, because of enzyme immaturity, the adrenal glands in the newborn infant might fail to provide as adequate a reserve of function under conditions of stress as is shown by adults.…”