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Intravenous injections of adrenocorticotrophic hormone (ACTH) caused adrenal ascorbic acid depletion in rats in which the mobilization of endogenous ACTH had been prevented by pretreatment with large doses of deoxycorticosterone acetate (DCA). The fall in adrenal ascorbic acid concentration was dependent upon the dose of ACTH administered. This observation has been developed to give a sensitive method for the bioassay of ACTH in which the need for hypophysectomy is obviated.