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An in-situ continuous infusion technique was used to study the biogenesis of androgens and cortisol in the guinea-pig adrenal gland after the infusion of ³H- and ^14C-labelled precursors of these compounds. Pregnenolone was converted to 17-hydroxypregnenolone, progesterone, 17-hydroxyprogesterone and cortisol. The results of infusion of a combined dose of [³H]17-hydroxypregnenolone and [¹⁴C]progesterone indicated that the major pathway to cortisol is via 17-hydroxypregnenolone. No conclusive proof could be obtained regarding synthesis of dehydroepiandrosterone or androst-4-ene-3,17-dione. The only androgen detected in the guinea-pig adrenal venous blood was 11β-hydroxyandrost-4-ene-3,17-dione. The major and minor biosynthetic pathways involved in the synthesis of these hormones are discussed.