IDENTIFICATION OF CORTICOSTEROIDS OF BEEF ADRENAL EXTRACT BY PAPER CHROMATOGRAPHY

Abstract

Chemical studies initiated in the laboratories of Reichstein, Kendall, and Wintersteiner (1) resulted in the isolation of twenty-eight crystalline steroids from adrenal gland extracts. Twelve of the compounds possessed an a-ketol side chain and, of these, six proved to be biologically active. The finding of 11-desoxycorticosterone, one of these active steroids, by Reichstein and von Euw (2) has been corroborated neither in the other two laboratories nor by subsequent workers. The methods used by these workers for the fractionation of the extracts and isolation of the crystalline compounds were such that the possibility of chemical alteration of some of the more labile steroids could not bi excluded. These methods, in addition, gave no reliable data as to the quantitative relationships existing among the different compounds found in the gland. The recent development in our laboratories of paper chromatographic and spectrophotometric techniques for the identification of adrenocortical steroids has been reported (3, 4). A procedure for the fractionation of complex corticoid mixtures and a systematic method for the identification of the separated compounds based upon these techniques are herein described. These methods offer several advantages over those used by previous investigators: (1) Samples are subjected to a minimum of handling; (2) components present in very low concentrations can be isolated and identified; and (3) compounds very closely related in chemical structure can be separated and differentiated. Analysis of a commercial preparation of beef adrenal extract thereby indicated the presence of at least twelve cr-ketolic steroids, of which eight, including 11-desoxycorticosterone, have been identified. Of the remaining four, two may represent new compounds. A preliminary report of these findings has already been made (5).