Hemolysis interference in measuring fish plasma biochemical indicators.

The present study aimed to determine hemolysis interference in measurement of plasma biochemical parameters in fish specimens. For this purpose, blood samples were harvested from 24 Huso huso juveniles. After centrifugation, each plasma sample was divided into seven portions to make seven levels of hemolysis. Hemolysis was induced by addition of different percentages of the corresponding whole blood [0 (non-hemolyzed control group), 1, 3, 5, 10, 15, and 20% of whole blood]. Albumin, total protein, calcium, phosphorus, glucose, sodium, potassium, chloride, alkaline phosphatase (ALP), aspartate transaminase (AST), and alanine transaminase (ALT) were measured in different samples. Results showed that plasma albumin, ALP, potassium, and AST significantly increased (more than 200% at the highest hemolysis level) in the hemolyzed samples. Also, plasma total protein and phosphorus showed significant elevation in the hemolyzed samples (more than 70% at the highest hemolysis level). Plasma glucose, calcium, chloride, and ALT showed narrow but significant increase in hemolyzed samples (11.8-35.2% at the highest hemolysis level). Plasma sodium showed no significant changes in the hemolyzed samples. In conclusion, the present results show that hemolysis markedly affects plasma parameters levels, which interferes with plasma results interpretation. Therefore, analysis of hemolyzed samples should be avoided or the results should be interpreted with caution. This study encourages further investigations to develop methods for omitting the effects of hemolysis by sample blanking and/or presenting correction coefficients for measurement of plasma parameters in samples with different levels of hemolysis.