Acetaminophen undergoes toxic conversion in the liver to a free-radical intermediary which binds to glutathione. N-Acetylcysteine acts as a glutathione precursor when natural stores are depleted, and is an effective antidote for acetaminophen overdose. Mushrooms containing amatoxins (such as Amanita phalloides) may undergo similar toxic conversion. However, in our amatoxin-poisoned mouse model, N-acetylcysteine (1.2 g kg−1) produced no change in survival or hepatic enzyme elevation compared to control animals. We conclude that N-acetylcysteine has no clinical role in the treatment of Amanita phalloides ingestion.