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Objective: To evaluate the effect of iron supplementation on serum copper and zinc levels., Method: In a randomized, double-blind, placebo-controlled trial, 66 pregnant women with hemoglobin 13.2 g/dL or greater between the 13th and 18th week of pregnancy were randomized into case and control groups. From the 20th week until the end of pregnancy the case group received one ferrous sulfate tablet containing 50 mg elemental iron daily, while the control group received placebo. Hemoglobin, ferritin, copper, and zinc levels at 24-28 and 32-36 weeks of pregnancy were measured and compared., Results: In the case group, serum copper levels in the second and third trimester were significantly lower than the control group (P<0.01 and P<0.001, respectively). Serum zinc levels in the case group in the second and third trimester were also significantly lower than the control group (P<0.001)., Conclusion: Iron supplementation in pregnant women with hemoglobin greater than 13.2 g/dL reduces serum levels of copper and zinc.