Stable isotope absorption studies of iron have been limited by the high cost and limited availability of isotope ratio analysis using thermal ionization MS (TIMS). The development of high-resolution double focusing inductively coupled plasma MS (ICP-MS) may permit more cost-efficient sample analysis due to its high throughput, lower cost, easy sample pretreatment, and greater availability. Our objective was to develop an ICP-MS methodology for the measurement of iron isotope ratios using very small blood volumes. We developed a technique using multiple iron-nickel mixing standard solutions to adjust for nickel interference calibration. RBC samples from human subjects previously given 58Fe and 57Fe were analyzed for iron isotope ratios and compared with our current methodology (TIMS). Reproducibility of iron isotope ratios provided external relative SD < 0.5 and 0.7% (1 SD) for 57Fe/54Fe and 58Fe/54Fe, respectively. Iron isotope ratios from ICP-MS analysis did not differ from those from TIMS based on statistical analyses, nor did the calculated iron absorption values. The mean and SD of iron absorption did not differ when measured by TIMS or ICP-MS. A 2-microL RBC sample was sufficient for ICP-MS iron isotope ratio analysis with an internal relative SD < 0.5% and analytical time < 5 min. This technique may assist groups in increasing their use of stable isotope methods to assess iron absorption in infants and children.