The Normal human pregnancy is associated with maternal adrenocortical hyperfunction that apparently is secondary to increased corticotropin (ACTH) secretion, since concentrations of maternal plasma ACTH increase progressively during pregnancy; urinary excretion of free cortisol is relatively resistant to glucocorticoid negative-feedback suppression (1, 2). It has been suggested that the placenta is involved in this increased secretion of ACTH because several workers have reported the presence of bioactive (1-8) or immunoreactive ACTH (1-3) in placental extracts. The ACTH extracted from placenta appears to consist, in addition to ACTH, of a high Mr form of the hormone (3). In mouse pituitary tumor cells (9, 10) and human nonpituitary carcinoma cells (11,12), ACTH appears to be formed by processing of a high Mr precursor molecule. This ACTH precursor contains the 13-lipotropin (f3LPH) sequence and, therefore, may also be processed to form f3LPH and its related peptides, yLPH [(1-58)/3LPH] and f3-endorphin [BEND,/3LPH] (9,11,12). The high Mr ACTH precursor molecule thus contains antigenic determinants for ACTH, the LPHs, and BEND.In the present study, we have used radioimmunoassays for ACTH, LPH (J3LPH plus "yLPH), 'yLPH, and fiEND, gel exclusion chromatography, affinity chromatography, sodium