The calcium content of human erythrocytes.

1. The calcium content of human erythrocytes, after removal of the buffy coat and washing free from plasma with isotonic sodium chloride, has been determined by atomic absorption spectrophotometry. The mean value found for normal subjects was 0.634 mug/ml. of packed erythrocytes (0.0158 mug-atom/ml.). The corresponding values for magnesium and zinc were 79.7 and 20.1 mug/ml., respectively.2. The calcium is considered to be mostly and perhaps exclusively located in the erythrocyte membrane, since, after osmotic haemolysis, the same amount was found in the ghost cells as was present in the erythrocytes from which they were prepared. By contrast, magnesium and zinc, which are essentially intracellular, were lost to the extent of about 96 and 92%, respectively.3. About 90% of the calcium was removed from erythrocytes by washing with isotonic sodium chloride containing 5 mM ethylenediaminetetraacetate (EDTA), or other complexing agents of high stability constant for calcium. A small fraction of the magnesium but none of the zinc was removed by this treatment.4. Other complexing agents of lower stability constant removed somewhat less calcium from the erythrocytes. Citrate was totally ineffective.5. The buffy coat had a high calcium content, but this could not be removed by washing with EDTA.6. Calcium was also determined in trichloroacetic acid extracts of ghost cells after ashing and treatment with bis-(o-hydroxyphenylimino)-ethane and measuring the red complex spectrophotometrically. The values obtained confirmed the atomic absorption measurements.