Interference by hemolysis with various methods for total calcium and its correction by trichloroacetic acid precipitation.

We evaluated the influence of hemolysis on total serum calcium as determined with the Du Pont aca, SMA 12/60, Ektachem 400, Corning 940 EGTA titrator, and the Beckman Astra 8, comparing results with those obtained by atomic absorption spectroscopy. We find that hemoglobin does not influence calcium measurements with the SMA 12/60, Ektachem 400, or Beckman Astra 8. The presence of hemoglobin exceeding 2 to 3 g/L caused falsely high results with the aca and falsely low results with the Corning 940 titrator. Similar interferences may be observed with other titrating or colorimetric procedures that involve direct reaction of the sample with o-cresolphthalein complexone or calcein. Upon removal of the hemoglobin by precipitation with trichloroacetic acid, the values obtained with the aca and the Corning 940 EGTA titrator were similar to those measured by atomic absorption. With nonhemolyzed serum samples the acid treatment had little or no effect on the aca procedure but resulted in a positive bias of approximately 10% with the EGTA procedure. Thus we recommend this trichloroacetic acid procedure for measuring calcium in hemolyzed samples with the aca and, with certain reservations, with the EGTA titrator.