During the analysis of human urines for their steroid content, a substance was encountered which could not be identified with a known steroid. It was noticed in both laboratories; as a contaminant of the pregnanediol fraction from the urine of a female child with hirsutism (A.P.W.) and on two-dimensional paper chromatograms of urine extracts from five cases of congenital adrenal hyperplasia (R.W.H. E.). Acetylation and chromatography of the pregnanediol fraction showed that this substance was not pregnanediol.
The substance has since been found in almost all urines examined, childrens' and adults', in amounts ranging from a few micrograms to several milligrams/24 hr., but as yet the excretion cannot be related to age, sex, or physiological state. Generally it is excreted as the glucuronide, but is sometimes unconjugated.
Isolation was achieved by ethyl acetate extraction following enzyme hydrolysis of extracted urinary glucuronides, separation into ketones and non-ketones with Girard's reagent, and
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