The adrenal cortex shows a histological and functional zonation which allows it to function as two distinct glands. Aldosterone is produced only in the outer area, or zona glomerulosa and cortisol is produced in the zona fasciculata. Adrenal slices (200 mu) were prepared from cores of beef adrenals using a McIlwain Tissue Slicer and incubated. Cortisol, corticosterone, aldosterone, 18-hydroxycorticosterone, 18-hydroxycortisol and 18-oxocortisol were measured by radioimmunoassay. Aldosterone and 18-hydroxycorticosterone were produced primarily in the outer slices of the adrenal. Cortisol and corticosterone were produced throughout the adrenal with the production of cortisol in the first layer probably due to contamination with fasciculata. 18-Hydroxycortisol and 18-oxocortisol were produced primarily in the outer slices. The coexistence of cytochrome P-450 corticosterone methyl oxidase I and II and the 17 alpha-hydroxylase in the outer slices leads to the synthesis of these two hybrid steroids.