Pulsed electric field (PEF) treatment (35 kV cm(-1) for 19.2 μs using bipolar 2 μs pulses) was conducted on bovine lactoferrin (LF; 0.4 mg mL(-1)) prepared in simulated milk ultrafiltrate (SMUF), at concentrations between 0.2× and 2× normal strength, with electrical conductivities ranging from 0.17 to 1.04 S m(-1). The physicochemical and structural characteristics (LF content by a spectrophotometric and an ELISA method, surface hydrophobicity, electrophoretic mobility, far-UV circular dichroism spectra, and tryptophan fluorescence) of LF dissolved in SMUF of all strengths tested were not changed after PEF treatment. The PEF treatment of LF in 0.2 strength SMUF did not cause the release of LF-bound ferric ion into the aqueous phase, with a concentration of LF-bound iron being the same as that of the untreated LF control (174 μg L(-1)). However, in treatment media with higher ionic strengths, ferric ion was released from the LF molecule into the aqueous phase. The concentration of LF-bound iron decreased from 174 μg L(-1) for the LF treated in 0.2 strength SMUF to 80 μg L(-1) for that treated in double-strength SMUF. The results suggest that the PEF-induced iron depletion of LF does not appear to cause an appreciable conformational change in LF molecules. PEF treatment could be developed as a novel physical way to produce iron-depleted LF, as an alternative to the existing chemical method.