Demonstration of aldosterone and corticosterone as the principal steroids formed in incubates of adrenals of the American bullfrog (Rana catesbeiana) and stimulation of their production by mammalian adrenocorticotropin

Aldosterone and corticosterone were found to be the only Δ⁴-3-ketosteroids produced to any significant extent in vitro by adrenal tissue from the American bullfrog. The production of aldosterone was stimulated highly by bovine adrenocorticotropin as well as by the frog's own anterior pituitaries. The amount of aldosterone produced by the bullfrog adrenal tissue under the stimulation of highly purified bovine ACTH was about four times the amount of corticosterone.The material from which the steroids were isolated was purified by solvent partitioning and reversed phase chromatography on purified acetylated filter paper according to a newly developed method. The identity of the isolated material with aldosterone was confirmed by the location of the ultraviolet absorption maximum at 239 mμ, by the fact that a characteristic spectrum was obtained before and after heating in sulfuric acid, on the basis of a positive reaction with blue tetrazolium and of the absence of any reaction with the Porter-Silber reaction, and by the results of bioassay in the Na²⁴-K⁴² test.