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Clinical Chemistry 13: 855-876, 1967;
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Clinical Chemistry, Vol 13, 855-876, Copyright © 1967 by the American Association for Clinical Chemistry

Sequential Solvent Extraction of Urinary Corticosteroids and its Clinical Application

Habeeb Bacchus 1

1 Metabolism Laboratory and Department of Pathology, Providence Hospital, Washington, D. C. 20017.

A procedure which permits the measurement of corticosteroid metabolites in three solvent extracts of hydrolyzed urine is described. Carbon tetrachloride extracts the 11-desoxy, 17-hydroxycorticosteroids, and the 17-OH, 21-deoxycorticosteroids. These are quantitated by the Porter-Silber reaction and the method for 17-ketogenic steroids. The difference between 17-KGS and PSC is a measure of the total 17-OH, 21-deoxycorticosteroids. Subsequent treatment of the residual urine with methylene dichloride extracts compounds of the cortisol series, i.e., cortisol and cortisone in small amounts, THF and THE in larger amounts. These compounds are quantitated by either the Porter-Silber reaction or the 17-KGS procedure. Further treatment of the urine residue with ethyl acetate (employing the "salting out" procedure) results in the extraction of polar compounds including cortol, cortolone, 6 -hydroxycortisol, and its reduction product. This procedure was employed to study several clinical disorders.

Congenital virilizing adrenocortical hyperplasia is characterized by a marked increase in the 17-KGS in the carbon tetrachloride extract; in adult-onset virilizing adrenocortical hyperplasia this increase is also significant, but not as marked. In adrenal hyperactivity (e.g., Cushing's syndrome, ACTH infusion) there are increases in all solvent fractions studied. Inhibition of 11- hydroxylase activity induced by SU 4885 (Metopirone) is accompanied by an increase in the PSC in the carbon tetrachloride fraction. In hepatocellular disease and in pregnancy, the most noteworthy changes are in the increase in polar compounds extracted in the ethyl acetate fraction. The method described is attractive in its simplicity and is readily applicable in the clinical laboratory.

Submitted on February 27, 1967
Accepted on April 13, 1967






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Copyright © 1967 by the American Association for Clinical Chemistry.