BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 234, 269?274 (1997)
ARTICLE NO. RC976622
Electrolyzed-Reduced Water Scavenges Active Oxygen Species and Protects DNA from Oxidative Damage
Sanetaka Shirahata,1 Shigeru Kabayama, Mariko Nakano, Takumi Miura,
Kenichi Kusumoto, Miho Gotoh, Hidemitsu Hayashi,* Kazumichi Otsubo,**
Shinkatsu Morisawa,** and Yoshinori Katakura
Institute of Cellular Regulation Technology, Graduate School of Genetic Resources Technology, Kyushu University, 6-10-1
Hakozaki, Higashi-ku, Fukuoka 812-81, Japan; *Water Institute, Nisshin Building 9F, 2-5-10 Shinjuku, Tokyo 160,
Japan; and **Nihon Trim Co. Ltd., Meiji Seimei Jusou Building 6F, 1-2-13 Shinkitano, Yodogawa-ku, Osaka 532, Japan
Received March 21, 1997
Active oxygen species or free radicals are considered to cause extensive oxidative damage to biological maccataromolecules, which brings about a variety of diseases as well as aging. The ideal scavenger for active oxygen should be ‘active hydrogen’. ‘Active hydrogen’ can be produced in reduced water near the cathode during electrolysis of water. Reduced water exhibits high pH, low dissolved oxygen (DO), extremely high dissolved molecular hydrogen (DH), and extremely negative redox potential (RP) values. Strongly electrolyzed–reduced water, as well as ascorbic acid, (/)-catechin and tannic acid, completely scavenged O•02 produced by the hypoxanthine-xanthine oxidase (HX-XOD) system in sodium phosphate buffer (pH 7.0). The superoxide dismutase (SOD)-like activity of reduced water is stable at 47C for over a month and was not lost even after neutralization, repeated freezing and melting, deflation with sonication, vigorous mixing, boiling, repeated filtration, or closed autoclaving, but was lost by opened autoclaving or by closed autoclaving in the presence of tungsten trioxide which efficiently adsorbs active atomic hydrogen. Water bubbled with hydrogen gas exhibited low DO, extremely high DH and extremely low RP values, as does reduced water, but it has no SOD-like activity. These results suggest that the SOD-like activity of reducedwater is not due to the dissolved molecular hydrogen but due to the dissolved atomic hydrogen (active hydrogen). Although SOD accumulated H2O2 when added to the HX-XOD system, reducedwater decreased the amount ofH2O2 produced by XOD. Reduced water, as well as catalase and ascorbic acid, could directly scavenge H2O2 . Reduced water suppresses single-strand breakage of DNA by active oxygen species produced by the Cu(II)-catalyzed oxidation of ascorbic acid in a dose-dependent manner, suggesting that reduced water can scavenge not only O•02 and H2O2 , but also 1O2 and •OH. q 1997 Academic Press
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