The major hydroxylated metabolite of leukotriene B4 in rat PMNL was found identical (UV spectrum and retention times in 3 different HPLC systems) to a synthetic compound of known stereochemistry, 19-hydroxy-LTB4. PMNL from various species exhibited 3 different types of behaviour for LTB4 hydroxylation. Human and monkey PMNL showed a high hydroxylating activity and a high regioselectivity with almost exclusive formation of products from 20-hydroxylation. Rat and mini-pig PMNL exhibited a very different regioselectivity with major formation of 19-OH-LTB4 (3:1 ratio). Finally, pig and beef PMNL were found almost devoid of any hydroxylating activity toward LTB4.