Ingestion of amniotic fluid and placenta by rats has been shown to enhance opioid-mediated antinociception but not affect the nonopioid-mediated antinociception produced by aspirin, suggesting specificity for opioid-mediated processes. However, enhancement by the active substance(s) in amniotic fluid and placenta (POEF, for placental opioid-enhancing factor) of antinociception produced by other nonopioid mechanisms has yet to be examined. The present experiments tested whether ingestion of amniotic fluid enhances the antinociception produced by nicotine injection. In Experiment 1A, enhancement of morphine-mediated antinociception by ingestion of amniotic fluid was demonstrated in a hot-plate assay. In Experiment 1B, rats pretreated with naltrexone were given an orogastric infusion of amniotic fluid or control (0.25 ml), then injected with nicotine (0, 0.075, 0.125, or 0.225 mg/kg subcutaneously), then tested for antinociception in a hot-plate assay. Amniotic fluid ingestion did not enhance the antinociception produced by various doses of nicotine. In Experiment 2, rats pretreated with naltrexone were given an orogastric infusion of amniotic fluid (0, 0.125, 0.25, or 0.50 ml) and then injected with 0.125 mg/kg nicotine. None of the doses of amniotic fluid enhanced the nicotine-induced antinociception. The findings of these experiments lend support to our contention that the enhancement by POEF of antinociception is specific to opioid-mediated processes.