Elevated iron stores may or may not promote atherogenesis by increasing free radical formation and oxidative stress, but controlled diet and supplement trials are lacking. We tested the hypothesis that iron supplementation does not increase the susceptibility of LDL to undergo oxidative modification in women with low iron status. A randomized, double-blind, 2-period crossover study design (n=26) was used to examine the effects of the following diets on measures of LDL oxidation: average American diet (AAD) [36% of energy as fat; 15% saturated fatty acids (SFA)], and a Step 2 diet (26% fat; 7% SFA). In addition, subjects received either a supplement containing 160 mg of ferrous sulfate (50 mg elemental iron) or a placebo twice daily [supplement group received a total of 320 mg ferrous sulfate (100 mg elemental iron) daily]. After supplementation, serum ferritin differed between the supplement and placebo groups (P=0.008). Measures of LDL oxidation were not affected by supplement intake; however, they were affected by diet. Lag time was shorter after the women consumed the AAD diet than after the Step 2 diet (P<0.0001). The diets did not affect the rate of oxidation or total dienes. Although iron status was improved by aggressive iron supplementation, LDL oxidative susceptibility was not affected. As expected, lag time was increased after the women consumed the low fat, low SFA diet. Therefore, the results of this study do not support a relationship between iron status and LDL oxidation.