Infusion medium containing a tracer was introduced through the left uterine artery to 20.5-day pregnant rats. In this way, the left uterine horn received the tracer directly while it reached the right horn after dilution in the mother’s circulation. When U-14C-D-glucose or U-14C-L-alanine was infused, radioactivity in fetal blood from the left uterine horn was 3 or 8 times higher respectively than in the right horn and 2 or 6 times higher than in maternal blood. When the infusion was carried out with 125I-labelled growth hormone, which is known not to cross the placental barrier, very little radioactivity was found in fetuses compared with that in the mother’s plasma and no differences were observed in blood radioactivity levels of fetuses from either horn. After infusion of U-14C-D)-glucose through the maternal jugular vein instead of the left uterine artery, radioactivity was the same in fetuses from the left and right horns, indicating that manipulations of the left uterine side did not greatly affect its circulation. These findings, together with the determination of specific radioactivity of the tracer used in both maternal and fetal plasma, validate the technique as a semiquantitative means to determine the direct transfer of any product through the placental barrier in the rat.