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ADRENAL steroid activity has been demonstrated in adrenal venous blood by means of biologic assay (1, 2). The biologic techniques, although successful in demonstrating the presence of adrenal steroids, have been unsatisfactory from a quantitative standpoint and have given little information regarding the specific compound being measured. Chemical methods for the determination of adrenal steroids in blood have been proposed by Corcoran and Page (3), and by Porter and Silber (4). The method of Corcoran and Page was found to give high values due to the formation of formaldehyde from phospholipids, traces of which could not be eliminated even by repeated precipitation with acetone. The color reaction of Porter and Silber depends on the formation of phenylhydrazones in acid solution. Under the conditions given, the 17,21-dihydroxy-20-ketosteroids showed a strong maximum absorption at 410 mμ when several steroid ketones were tested. The purification procedure used with plasma, however, was so nonspecific as to raise the question of the influence of other compounds on the reaction.
Nelson et al. (1952) studied this question.