A reevaluation of the chemotactic potency of leukotriene B4 (LTB4)

The chemotactic potency of leukotriene B4 (LTB4) was reevaluated based on an improved purification procedure which combines reversed phase and straight phase high pressure liquid chromatography (RP-HPLC and SP-HPLC). Socalled LTB4 isomer III prepared by RP-HPLC contains two double oxygenated 5,12-dihydroxy acids in addition to LTB4. On a molar basis, the chemotactic activity of LTB4 repurified by SP-HPLC was far greater than that of the other two 5,12-dihydroxy acids and comparable to that of formyl-methionyl-leucyl-phenylalanine (fMLP). The chemotactic activity of LTB4 isomer III is dependent upon the relative concentrations of the double oxygenated 5,12-dihydroxy acids and LTB4. Further purification of peak III by SP-HPLC is required before assessing the biologic activity of LTB4.