The effect of calcium supplement use on iron stores was examined in a randomized controlled study in free-living, healthy, premenopausal women. Of 109 women who completed the study, 52 were in the control group and 57 took two tablets containing 250 mg Ca as the carbonate with each of two meals daily for 12 wk. In all subjects at baseline, plasma ferritin concentrations were positively correlated with heme-iron intake (r = 0.21, P = 0.04), serum iron concentration (r = 0.19, P = 0.04), transferrin saturation (r = 0.31, P = 0.001), and hemoglobin concentration (r = 0.22, P = 0.02), and negatively correlated with total iron-binding capacity (TIBC, r = -0.42, P < 0.001). No significant differences in absolute or percent changes in plasma ferritin concentrations, serum iron concentrations, TIBC, transferrin saturation, hemoglobin concentrations, or hematocrit were observed between the treatment and control groups. Thus, over a 12-wk period, use of 1000 mg Ca as the carbonate daily with meals does not appear to be detrimental to iron stores in healthy, free-living, premenopausal women.