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, Tokko 160, Japan; and **Nihon Trim Co. Ltd.,
Meiji Seimei Jusou Building 6F, 1-2-13
Shinkitatno, Yodogawa-ku, Osaka 532, Japan
Received March 21. 1997
Active oxygen species or free radicals are considered to cause extensive oxidative damage to biological macromolecules, 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·2 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 4º C 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 reduced water 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, reduced water decreased the amount Of H202 produced by XOD. Reduced water, as well as catalase and ascorbic acid, could directly scavenge H202. 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·2 and H2O2, but also ·O, and ·OH. – 1997
Strong electrolyzed-reduced water completely scavenged 02 produced by the hypoxanthine-xanthine oxidase (HX-XOD) system at pH 7.0 Although SOD accumulated h202 when added to the HX-XOD system, reduced water decreased the amount of h202 produced by XOD. Reduced water suppressed single-strand breakage of DNA by active oxygen species produced by the Cu(II)-catalyzed oxidation of ascorbic acid. The expression of Interleukin-6 gene was enhanced by reduced water as well as ascorbic acid, (+)-catechin and tannic acid when added to the culture of human osteosarcoma cell line, MG-63 cells, suggesting that reduced water acts as a reductant to cells.
RESULTS AND DISCUSSION
Characteristics of electrolyzed-reduced water. The principle of electrolysis was founded by Michael Faraday (1791-1867). In this process, reduction occurs at the cathode (+) and oxidation at the anode (-). Dissociation of H2O produces H- and OH- ions. At the cathode, H+ ions gain electrons to change into active atomic hydrogen (H-) Active atomic hydrogen exhibits high reducing potential. It is then changed to hydrogen molecules (H2) which are chemically inert at room temperature. At the anode, OH- ions lose electrons to form OH, which results in the production of O2, and H2O. Cathodic alkaline water (reduced water) is abundant in DH, whereas anodic acidic water (oxidized water) is abundant in DO. The relationships of RP with pH, DO, and DH in reduced water were shown in FIG. 1. Marked changes in these values occur in water after electrolysis. It should be noted that the DH value is higher in reduced water than in the original water by two orders of magnitude.