Electrochemically reduced water exerts superior reactive oxygen species scavenging activity in HT1080 cells than the equivalent level of hydrogen-dissolved water

PLoS One. 2017 Feb 9;12(2):e0171192. doi: 10.1371/journal.pone.0171192. eCollection 2017.


Electrochemically reduced water (ERW) is produced near a cathode during electrolysis and exhibits an alkaline pH, contains richly dissolved hydrogen, and contains a small amount of platinum nanoparticles. ERW has reactive oxygen species (ROS)-scavenging activity and recent studies demonstrated that hydrogen-dissolved water exhibits ROS-scavenging activity. Thus, the antioxidative capacity of ERW is postulated to be dependent on the presence of hydrogen levels; however, there is no report verifying the role of dissolved hydrogen in ERW. In this report, we clarify whether the responsive factor for antioxidative activity in ERW is dissolved hydrogen. The intracellular ROS scavenging activity of ERW and hydrogen-dissolved water was tested by both fluorescent stain method and immuno spin trapping assay. We confirm that ERW possessed electrolysis intensity-dependent intracellular ROS-scavenging activity, and ERW exerts significantly superior ROS-scavenging activity in HT1080 cells than the equivalent level of hydrogen-dissolved water. ERW retained its ROS-scavenging activity after removal of dissolved hydrogen, but lost its activity when autoclaved. An oxygen radical absorbance capacity assay, the 2,2-diphenyl-1-picrylhydrazyl assay and chemiluminescence assay could not detect radical-scavenging activity in both ERW and hydrogen-dissolved water. These results indicate that ERW contains electrolysis-dependent hydrogen and an additional antioxidative factor predicted to be platinum nanoparticles.

MeSH terms

  • Cells, Cultured
  • Electrolysis*
  • Free Radical Scavengers / chemistry
  • Free Radical Scavengers / pharmacology*
  • Humans
  • Hydrogen / analysis
  • Hydrogen / chemistry*
  • Hydrogen / pharmacology
  • Hydrogen Peroxide / pharmacology
  • Oxidation-Reduction / drug effects
  • Oxidative Stress* / drug effects
  • Reactive Oxygen Species / metabolism*
  • Water / chemistry*
  • Water / physiology


  • Free Radical Scavengers
  • Reactive Oxygen Species
  • Water
  • Hydrogen
  • Hydrogen Peroxide

Grants and funding

This work was supported in part by Nihon-Trim Co., Ltd and Kyushu university's finance (Trust Accounts No. JAKF650803). Shigeru Kabayama is employed by Nihon-Trim Co. Ltd. Nihon Trim Co. Ltd provided support in the form of salary for author SK, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific role of this author is articulated in the ‘author contributions’ section.