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A variety of ¹⁴C and ³H-labelled steroids have been perfused through the human adrenal gland in situ and their metabolic products isolated from adrenal venous blood.

Progesterone, dehydroepiandrosterone and cortisol were isolated after infusion of [³H]pregnenolone; 17-hydroxyprogesterone, dehydroepiandrosterone and cortisol after infusion of [³H]17-hydroxypregnenolone and [¹⁴C]progesterone; androstenedione and cortisol after infusion of [³H]17-hydroxyprogesterone and [¹⁴C]dehydroepiandrosterone; and 11β-hydroxyandrostenedione after infusion of [³H]androstenedione and [¹⁴C]cortisol.

From a consideration of the incorporation of radioactivity into the metabolic products, the³H: ¹⁴C ratios and the tissue pool sizes it was concluded that the major biosynthetic pathway to cortisol in the human adrenal glands was: pregnenolone 17-hydroxypregnenolone cortisol. Progesterone was not an important intermediary.

Androstenedione was mainly formed by way of 17-hydroxypregnenolone 17-hydroxyprogesterone androstenedione. 11β-Hydroxyandrostenedione was formed mainly from cortisol and only a minor amount came from the hydroxylation of androstenedione.