Leukotriene B4 (LTB4) has been implicated as a mediator in the inflammatory process by virtue of its potent chemotactic activity. At present, very little is known of the stability of this compound in vivo; therefore, the present study was designed to determine the half-life and metabolic fate of radiolabeled LTB4 during a 2-hr intrapleural incubation in rats with acute carrageenan pleurisy. After injection of 0.2 ml of 1% sodium carrageenan (Viscarin), inflammation was allowed to develop for 4 hr. A small polyethylene cannula was then inserted into the chest, and 0.1 microCi of [14C]LTB4 was injected into the chest. Samples for radioactivity determination were taken at 0, 1, 2, 3, 4, 5, 7, 10, 15, 20, 30, 45, 60, 90 and 120 min via the cannula, and at 120 min the entire content of the chest was collected. The half-life for the disappearance of radioactivity from the chest was 45.8 +/- 3.5 min. The 120-min samples were treated with acetone to precipitate protein and extracted with Sep-Paks. The extracts were analyzed by reversed phase high performance liquid chromatography using an ultraviolet detector set at 269 nm and a radioactivity detector. An additional experiment was run using multi-[3H]LTB4, and the only major metabolites detected were omega-hydroxylated compounds. It can be concluded from these results that LTB4 is relatively stable in vivo and could be present for long enough at the inflammatory site to have an influence upon inflammatory cell migration.