A Sensitive In vitro Spectrophotometric Hydrogen Peroxide Scavenging Assay using 1,10-Phenanthroline

Abstract

Introduction: Hydrogen peroxide (H₂O₂) is a biologically important, non-radical reactive oxygen species (ROS) that can influence several cellular processes. Ability of anti-oxidants to scavenge H₂O₂ can be measured by several methods but all the methods are suffering from several lacunae due to poor reproducibility, lack of specificity, inaccuracy, high cost etc. Objective: The present study aimed towards development of a rapid, low cost, reproducible, specific as well as sensitive and accurate method to detect H₂O₂ scavenging activity of anti-oxidants. Methods: We have used 1,10-phenanthroline and ferrous ammonium sulphate to detect H₂O₂ scavenging activity of anti-oxidants. Results: We revealed that our assay is able to detect the in vitro H₂O₂ scavenging activity of both phenolic and non-phenolic anti-oxidants. Furthermore, we found that this assay is highly specific, as scavengers of other types of ROS were unable to show any detectable effect on H₂O₂ through this assay. Finally, we tested commercially available non-steroidal anti-inflammatory drugs that are known to possess H₂O₂ scavenging activity and in such drugs also we were able to detect their H₂O₂ scavenging activity. Conclusion: In conclusion, the proposed spectrophotometric method was rapid, cost effective, reproducible and highly specific to detect in vitro H₂O₂ scavenging activity of diverse compounds.