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Updated: May 20, 2026
Methods for Quantitative Detection of Antibody-induced Complement Activation on Red Blood Cells
Published on: January 29, 2014
An analysis of the bias in red blood cell hemolysis measurement using several analytical approaches
Jason P Acker1, Ioana M Croteau, Qi-Long Yi
1Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB T6G 2R8, Canada. jason.acker@blood.ca
Background:
Percentage hemolysis in red cell concentrates (RCC) for transfusion is an indicator of RBC damage. As several factors need to be measured to determine hemolysis, and multiple assays are available for each, the choice of analytical methodology could critically influence results.
Methods:
Hemolysis was measured in 48 RCCs every 7 days during storage including expiry (42 days), with supernatant hemoglobin measured using the reference Drabkin's cyanmethemoglobin method and the Harboe spectrophotometric method, total hemoglobin measured using Drabkin's method and 3 automated analyzers (ADVIA 120, CELL-DYN 1700, Coulter AcT), and hematocrit measured using traditional centrifugation and automated analyzers.
Results:
The choice of method affects hemolysis measurement. Biases ranging from -0.01- -0.03% were observed depending on the combination of methods used. Hematocrit measurement appeared to be a major determinant of bias, and the greatest bias was seen with the ADVIA 120 automated analyzer. Although results did not differ by a level thought to be of clinical significance, the choice of method will impact quality control pass/fail rates.
Conclusion:
Although guidelines exist in many jurisdictions regarding acceptable hemolysis levels in RCCs, these are silent regarding the methods to be used. The presence of bias highlights the need for standardization of methodology.
