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NORMAN I. GOLD, JOHN F. CRIGLER, Evidence for 3α,17,21-Trihydroxy-5α-pregnane-11,20-dione (“ATHE”), a Metabolite of Cortisol in Urine of Infants, The Journal of Clinical Endocrinology & Metabolism, Volume 30, Issue 1, 1 January 1970, Pages 71–75, https://doi.org/10.1210/jcem-30-1-71
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Abstract
Cortisol and cortisone, one labeled with 3H and the other with 14C, were simultaneously administered iv to 6 infants, male and female, 2/3 to 16 months of age and to 1 boy 30 months of age. The radioactivity derived from the cortisol label was determined by reverse isotope dilution procedures for the following glucosiduronate metabolites of cortisol present in urine collected for 48 hr after labeled hormone administration: 3α,11β,17,21-tetrahydroxy-5β-pregnan-20-one (THF), 3α,17,21-trihydroxy-5β-pregnane-11,20-dione (THE), 3α,11β,17,21-tetrahydroxy-5α-pregnan-20-one (ATHF) and a new metabolite identified as 3α,17,21-trihydroxy-5α-pregnane-11,20-dione (ATHE). The presence of the additional label derived from cortisone in the metabolites and their oxidation products permitted calculation of the constancy of (14C SA)/cm and the slope of (3H/14C)/cm, due to the chromatographic 3H-isotope effect, across the metabolite peaks. This aided the definition and specificity of the chromatograpliic techniques. The relative amount of each metabolite was calculated as 100× (metabolite) dpm/(THF+THE+ATHF+ATHE) dpm. ATHE was present in the urine of 5 of the 6 infants in amounts, 8–24%, comparable to the quantities of THF, 1–19%, and ATHF, 5–28%. The following characteristics of the new metabolite were consistent with its identification as ATHE: a) chromatographic mobility, b) reaction with acid phenylhydrazine, c) transformation to a single product, 5α-androstane- 3,11,17-trione, with chromic acid oxidation, and d) transformation to a single product, 3α-hydroxy-5α-androstane-11,17-dione, upon oxidation with sodium bismuthate. The frequent occurrence of ATHE in the urine of infants and its relative absence in adults is another example of a difference in cortisol metabolism in infants.