The kinetics of iron removal from the two metal binding sites of the bovine lactoferrin by ethylenediaminetetraacetic acid (EDTA) was investigated at pH 7.5 and 33 °C. Solutions were buffered at pH 7.5 by 0.15 M Tris–HCl. Pseudo first-order rate constants as a function of ligand concentration were measured for iron removal from diferric lactoferrin and from N- and C-terminal monoferric lactoferrin. Diferric lactoferrin showed simple saturation behavior while both the monoferric forms showed a two-term dependence of k obs on ligand concentration that signifies two pathways for iron removal under the conditions applied. Moreover, the results show that the N-terminal site is more labile towards iron removal by EDTA than the C-terminal site.