Influence of Anticoagulants on Determination of H2O2 Levels in Blood: Comparison of Citrate and EDTA

Abstract

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.

Autophagy in Diabetes Pathophysiology: Oxidative Damage Screening as Potential for Therapeutic Management by Clinical Laboratory Methods

“…Diagnostic laboratory markers include the traditional risk evaluation markers such as cholesterol and glucose profiles. Emerging biomarkers include C-reactive protein, D-dimer, and homocysteine (Tracy, 2003 ; Ridker et al, 2004 ), as well as oxidative stress indices that include GSH and MDA (Tsimikas, 2006 ; Gyawali et al, 2015 ; Nwose et al, 2018 ; Lubrano et al, 2019 ). Uric acid and albumin levels have been considered (Bwititi et al, 2012 ).…”

Novel Platinum-Porphyrin as Sensing Compound for Efficient Fluorescent and Electrochemical Detection of H2O2

“…Reduced oxygen levels are developed in cardiac and brain ischemia [4]. Moreover, H 2 O 2 present in blood can interact with Fe 2+ ions contributing to oxidative stress [5]. system, on a Perkin Elmer LS55 luminescence spectrometer (Perkin Elmer, Inc./UK Model/LS 55, Waltham, MA, USA).…”