The concentrations of relaxin was determined by RlA [radioimmunoassay] in various compartments and tissues in pregnant women. Ten pairs of maternal venous and cord blood were studied. The mean relaxin concentrations, in immunoreactive equivalents of porcine relaxin, were 0.683 ng/ml in maternal serum and 0.009 ng/ml in cord serum. Relaxin was undetectable in 8 of 9 samples of amniotic fluid. Mean concentrations of relaxin, in pg immunoreactive equivalents of porcine hormone per mg protein, from maternal tissues at term pregnancy were: fat 93 (N = 5), myometrium 47 (N = 4), skin 62 (N = 5), placenta 51 (N = 9) and corpus luteum 13,000 (N = 8). Apparently little relaxin crosses the placenta and little is produced in the fetus. Contrary to prior suggestions that relaxin may also be a placental product, relaxin appears to be solely produced in the corpus luteum, making it the only peripherally measured hormone that can be used as an index of luteal activity in pregnancy.