An iron stable isotope comparison between human erythrocytes and plasma

We present precise iron stable isotope ratios measured by multicollector-ICP mass spectrometry (MC-ICP-MS) of human red blood cells (erythrocytes) and blood plasma from 12 healthy male adults taken during a clinical study. The accurate determination of stable isotope ratios in plasma first required substantial method development work, as minor iron amounts in plasma had to be separated from a large organic matrix prior to mass-spectrometric analysis to avoid spectroscopic interferences and shifts in the mass spectrometer's mass-bias. The Fe-56/Fe-54 ratio in erythrocytes, expressed as permil difference from the IRMM-014 iron reference standard (delta(56)/Fe-54), ranges from -3.1% to -2.2%, a range typical for male Caucasian adults. The individual subject erythrocyte iron isotope composition can be regarded as uniform over the 21 days investigated, as variations (+/- 0.059 to +/- 0.15%) are mostly within the analytical precision of reference materials. In plasma, delta(56)/Fe-54 values measured in two different laboratories range from -3.0 parts per thousand to -2.0 parts per thousand, and are on average 0.24 parts per thousand higher than those in erythrocytes. However, this difference is barely resolvable within one standard deviation of the differences (0.22 parts per thousand). Taking into account the possible contamination due to hemolysis (iron concentrations are only 0.4 to 2 ppm in plasma compared to approx. 480 ppm in erythrocytes), we model the pure plasma delta(56)/Fe-54 to be on average 0.4 parts per thousand higher than that in erythrocytes. Hence, the plasma iron isotope signature lies between that of the liver and that of erythrocytes. This difference can be explained by redox processes involved during cycling of iron between transferrin and ferritin.