CORTICOSTEROIDS IN HUMAN BLOOD .9. EVIDENCE FOR ADRENAL SECRETION OF SULFATE-CONJUGATED CORTISOL, 11-BETA, LPHA-DIHYDROXY-4-PREGNENE-3,20-DIONE-21-YL-SULFATE

A method was developed for the estimation of levels of cortisol-21-sul fate (FKS), cortisone-21-sulfate (E(K)S), and 20(alpha + beta)-reduced cortisol-21-sulfates in blood plasma. Levels of these conjugates were determined in peripheral vein plasma of 42 normal subjects, 21 men, a nd 21 women (age range 20-64 years) and in adrenal vein plasma of pati ents with various adrenocortical disorders, six patients with primary hyperaldosteronism, five patients with Cushing's syndrome, and in two obese patients, suspected to have Cushing's syndrome, but with inconcl usive laboratory findings. Adrenal vein blood was obtained by percutan eous, trans-femoral adrenal vein catheterization. Levels of non-conjug ated (free) cortisol were determined in all plasma samples along with those of the sulfated steroids. FKS was found in all plasma samples, b oth in men and women. The variation in FKS levels paralleled that in t he free cortisol levels, thus the ratio of F-K/FKS was the same in the blood samples drawn at 8 AM as in those drawn at 4 PM or 5 PM (ranges : 17.5-36.3 in men, 23.6-45.8 in women). The levels of FKS were relati vely lower in women than in men (women 610-880 ng/100 mt at AM, 300-51 0 ng/100 mL at PM; men: 760-1,220 ng/100 mL at AM, 380-760 ng/100 mL a t PM). Plasma levels of total sulfate-conjugated Delta(4)-3-keto-C-21 steroids (FKS + E(K)S + 20(alpha + beta)-dihydrocortisol-21-sulfates) were 30-40% higher than those of the levels of cortisol-21-sulfate alo ne (separated by thin-layer chromatography). In the adrenal vein plasm a, levels of Delta(4)-3-keto-C-21-steroid-21-yl sulfates were 20 to 40 times higher than levels of these steroids in the peripheral blood. T he bulk of the steroid sulfate measured in the adrenal vein plasma con sisted of cortisol-21-sulfate. The ratio of F-K/FKS in the adrenal vei n plasma was markedly smaller than in the peripheral vein plasma; it w as 6.9-12.3 in males and 4.9-6.7 in females, whereas in the peripheral vein of the same subjects it was 19.2-43.7 in males and 21.4-48.3 in females. Cortisol-21-sulfate isolated from adrenal vein plasma was ide ntified by mass spectrometry. The data presented provide evidence for the secretion of this conjugate by the adrenal cortex. Its secretion a ppears to be markedly elevated in patients with Cushing's syndrome, bo th due to hyperplasia and due to adrenal adenoma, as compared with nor mal subjects and patients with primary aldosteronism, both males and f emales. However, the F-K/FKS ratio was markedly lower in Cushing's pat ients due to adrenal adenoma than due to adrenal hyperplasia, this sug gesting that ACTH is stimulating intra-adrenal hydrolysis of cortisol sulfate.