Acetaminophen is a frequently prescribed over the counter drug to reduce fever and pain in the event of inflammatory process. As neutrophils are relevant cells in inflammatory processes, the putative interaction of acetaminophen with these cells, if present, would be of paramount importance. The present study was undertaken to evaluate the effect of acetaminophen in human neutrophils' oxidative burst and lifespan in vitro. The results of the present study demonstrate that acetaminophen efficiently modulates neutrophils' oxidative burst in phorbol myristate acetate-activated neutrophils, in a concentration-dependent manner, at in vivo relevant concentrations. It was clearly demonstrated that acetaminophen is a strong scavenger of HOCl and H2O2, which probably contributed to the effect observed in neutrophils, and depletion of glutathione in stimulated neutrophils, suggesting its transformation into a reactive intermediate. Obtained results also revealed that acetaminophen affects programmed cell death of human neutrophils, resulting in a delay of previously stimulated neutrophils-mediated apoptosis. Overall, our data suggested that acetaminophen has considerable potential as anti-inflammatory therapeutic strategy, by preventing biological damage induced by an excessive production of reactive species by activated neutrophils and by extending the lifespan of neutrophils, favoring the elimination of the offending insult, contributing this way to tissue healing and resolution of inflammation.