STUDIES ON ALDOSTERONE: I. ISOLATION FROM ADRENAL CORTICAL EXTRACT AND A TEST FOR ITS IDENTITY

Abstract

Details are given of a method which has proved very satisfactory for the isolation of aldosterone from commercial aqueous adrenal cortical extracts. The hormone was extracted with chloroform and separated by chromatography in the following systems: System I, benzene: 5% ethyl acetate/methanol: water; System II, toluene/propylene glycol; System III, benzene: 10% ethyl acetate/methanol: water. Identification of the isolated steroid was provided by: U.V. absorption, acetylation followed by chromatography, bio-assay, qualitative tests with blue tetrazolium and phenylhydrazine, absorption spectra in sulphuric and phosphoric acids, and crystallization. Quantitative estimations were made by U.V. absorption and the dinitrophenylhydrazine reaction. The latter method has been shown to exhibit with aldosterone a characteristic behavior that can assist in establishing the identity of this hormone. The yield from the extract principally employed was 4 μg. per ml.