Comparison of two methods for measuring free haemoglobin in the supernatant of erythrocyte transfusion products
Authors:
L. Řehořová 1; P. Matoušek 2; R. Procházková 1,3
Published in:
Transfuze Hematol. dnes,31, 2025, No. 1, p. 36-40.
Category:
Original Papers
doi:
https://doi.org/10.48095/cctahd2025prolekare.cz5
Overview
Introduction: During the storage of erythrocyte transfusion products, the level of free haemoglobin, a marker of ongoing haemolysis, increases. The European Council’s recommendations (Guide to the preparation, use, and quality assurance of blood components, 21st Edition) and Czech legislation (Decree No. 143/2008 Coll.) set the maximum level of free haemoglobin at 0.8% of erythrocyte mass by the end of the transfusion product’s shelf life. Measuring the percentage of haemolysis is also recommended for erythrocyte transfusion products that undergo further processing, such as washing. The aim of this study was to compare two methods for measuring free haemoglobin in the supernatant of erythrocyte transfusion products: direct spectrophotometry and the Harboe method for determining free haemoglobin content in transfusion products. Materials and Methods: The study included 20 units of erythrocyte transfusion products. At the end of the shelf life, supernatant samples were collected, and the free haemoglobin level was measured using direct spectrophotometry by determining the absorbance of the sample at 540 nm (A) and measuring absorbance at wavelengths of 415 nm, 450 nm, and 380 nm according to Harboe (B). The percentage of haemolysis of the erythrocyte mass was then calculated based on the volume and haematocrit of the erythrocyte transfusion product. Results: The level of free haemoglobin at the end of the storage period did not exceed 0.8% of erythrocyte mass for any unit – on average, 0.31% (A) and 0.29% (B). No statistically or clinically significant differences were found between the two methods. Conclusion: The method of measuring absorbance at wavelengths of 415 nm, 450 nm, and 380 nm according to Harboe corresponds to the commonly used method at 540 nm and is suitable for measuring free haemoglobin content in transfusion products.
Keywords:
Spectrophotometry – haemolysis – erythrocytes
Sources
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