We examined the effect of iron-containing prenatal vitamin-mineral supplements (PNV) taken postpartum on biomarkers of iron status and oxidative stress. Lactating women (n=114) were randomly assigned to consume daily one iron-free PNV plus either 27 mg of iron or placebo for approximately 3.5 mo. The placebo group took the tablets between meals, while those given iron took the tablets either with (Fe-W) or between meals (Fe-B). Blood and urine samples were collected before and after the supplementation period to analyze hemoglobin (Hb), ferritin, hepcidin, transferrin saturation (TfSat), total plasma iron, and biomarkers of oxidative stress (isoprostane and 8-hydroxy-2-deoxyguanosine (8-OHdG)) and inflammation (C-reactive protein (CRP) and alpha-1-acid glycoprotein (AGP)). There was a trend toward a greater change in Hb among women in the Fe-B group compared to placebo (+2.5 vs. -3.7 g/L, respectively, P=0.063). When the iron groups were combined, there was a greater change in Hb (+1.4 g/L) compared to placebo (P=0.010). There were trends toward greater changes in TfSat (P=0.087) and total plasma iron (P=0.065) in the iron groups compared to placebo, yet no significant differences between the 3 groups in change in hepcidin (P=0.291), isoprostane (P=0.319) or 8-OHdG (P=0.659), nor in change in ferritin among those with elevated CRP at baseline (60% of women; P=0.946); among those without elevated CRP (40% of women), ferritin increased more in the iron groups compared to placebo (P=0.001). Iron consumption during lactation moderately increased iron status, particularly among women without elevated CRP, and increased Hb, but did not significantly increase oxidative stress