Background: H 2 O 2 is supposed to be part of the laboratory test panel for assessment of oxidative stress. This molecule contributes to oxidative stress by reacting with Fe 2+ ions to produce hydroxyl radicals. Anticoagulants, including citrate and EDTA chelate Fe 2+ , possibly have implications in oxidative stress studies especially when H 2 O 2 is measured. Objective: This brief commentary is on the measurability of H 2 O 2 in citrated and EDTA blood samples. Method: Laboratory evaluation was performed using blood samples of 30 sheep collected in EDTA and citrate tubes. Samples were centrifuged, and plasma separated. H 2 O 2 was determined using the Biotech H 2 O 2 560 assay protocol on plasma. The ratio of EDTA to iron was also estimated to help interpret levels of H 2 O 2 in EDTA blood. Results: There was measurable H 2 O 2 reaction in citrate samples, but not in the EDTA blood. Based on brand sterile hematology tubes containing 1.8 mg EDTA per mL blood; approximately 4:1 ratio of EDTA to iron is estimated.Presuming this calculated ratio is a reflection of the true iron levels in the sheep's blood, the test results support the suggestion that ratio of EDTA to iron is greater than 1:1. Conclusion:Evidence is hereby presented to advance that EDTA anticoagulant is unsuitable for laboratory testing of H 2 O 2 in plasma. Further studies may be needed to ascertain the effects of different anticoagulants on H 2 O 2 in whole blood and haemolysate to see how these can be compared to plasma. This is relevant to establish a standard protocol for integration of H 2 O 2 test for assessment of full oxidative stress panel in clinical practice.
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