Optimized atomic absorption spectrophotometry of calcium in erythrocytes

We sought to establish optimum conditions for measuring calcium in erythrocytes by atomic absorption spectrophotometry. The conditions we selected are as follows. Wash one volume of fresh heparin-treated packed cells once with 30 volumes of isotonic buffered saline (pH 7.4) at a temperature somewhat exceeding 25 degrees C. Dilute the washed packed cells 10-fold with 12 mmol/L hydrochloric acid, and analyze the supernate for calcium. Measure the hematocrit of the washed packed cells, then analyze an aliquot of them for calcium, using a computer-readout type of flame or a non-flame atomic absorption spectrophotometer equipped with a pyrocoated graphite tube. The temperature program is 1000 degrees C for ashing [corrected] and 1800 degrees C for the atomizing cycle. Intraday and day-to-day reproducibility of the assay was 6.55% and 8.19%, respectively, at the mean concentration of calcium in the erythrocytes of healthy adults, which is 4.30 mumol/L.