Mechanisms responsible for anaphylactoid reactions to N-acetylcysteine (NAC) are poorly understood, and acetaminophen itself may play an important role. The present study examined the relationship between serum acetaminophen concentrations and risk of anaphylactoid reactions. Prospective study of adverse reactions to NAC administered according to standardized clinical protocols in patients who present to hospital after acute acetaminophen overdose. Subgroups were defined by serum acetaminophen concentrations 0 to 100 mg/L, 101 to 150 mg/L, 151 to 200 mg/L, 201 to 300 mg/L, and >300 mg/L. There were 362 patients, and anaphylactoid reactions occurred in 14.9%. Anaphylactoid reactions occurred less commonly in patients with high serum acetaminophen concentrations (p = 0.046 by Cochran-Armitage trend test) and high equivalent 4 h acetaminophen concentrations (p = 0.004). High serum acetaminophen concentrations were associated with fewer anaphylactoid reactions, suggesting that these might in some way be protective. The biological basis needs further exploration so as to allow a better understanding of the mechanisms responsible for adverse reactions to NAC treatment.