The in vitro effects of certain adrenal steroids and synthetic analogs on hepatic gluconeo-genesis have been investigated. Physiologic amounts of these hormones caused increased carbohydrate synthesis when liver of normal or adre-nalectomized rats was incubated with alanine. The steroid effect on gluconeogenesis was more marked in normal hepatic tissue. Incubation of liver slices with pyruvate resulted in greater formation of carbohydrate than when alanine was substrate; however, the steroids did not stimulate gluconeogenesis from pyruvate.