Corticosteroids in urine of normal persons determined by paper chromatography.

The presence in human urine of small amounts of material with adrenocortical activity is well established (1, 2). The chemical characterization of this urinary material by the classical methods used for the steroids in adrenal glands has been technically difficult. In the six reported instances (3-8) in which corticoids have been isolated and identified by these procedures, extracts of large volumes of urine containing quantities of corticosteroids of the order of several mg. were employed. These methods, furthermore, cannot be routinely used in the isolation of the corticosteroids present in conveniently small aliquots of urine. A paper chromatographic method for the separation of microgram quantities of adrenocortical steroids has already been reported (9). In the preceding paper (lo), the value of this procedure has been demonstrated in the separation of the components in small quantities of adrenal cortex, extract and a systematic method for the identification of these corticoid components, embracing both chromatographic and spectrophotometric techniques, has been described. The application of this method to the analysis of urinary extracts for their minute content of corticoids is complicated by the presence of very much larger quantities of pigments and other contaminating materials. The procedure devised for urine extracts is, consequently, more lengthy than that used in the analysis of adrenal extracts. This paper describes a method which requires relatively small samples of urine and can, therefore, be used in extensive studies of corticoid excretion patterns. Also given are the results of the analyses of urine specimens, each representing 3 days excretion, from normal persons, in which the presence of 17a-hydroxycorticosterone (Kendall’s Compound F) and of 11-dehydro-17~hydroxycorticosterone (cortisone) was demonstrated, as reported in preliminary communications (11, 12).