These preliminary investigations showed that the reduced hydrogen silicate mineral was an antioxidant (reducing agent) using several different methods. It was shown to directly reduce NAD+ to NADH effectively. NAD+ and its reduced form, NADH, are the primary cofactors participating in numerous biochemical reactions for metabolism and energy production in the cell including the mitochondria's electron transport chain and the coupled production of ATP (adenosine triphosphate). In the case of NAD-linked dehydrogenase enzymes such as glyceraldehyde-3-phos-phate dehydrogenase, reduced hydrogen (H:-) is generated which can directly enter the electron transport chain (11). Two reduced hydrogen ions are generated from a complete cycle of glycolysis with both these donating a total of four electrons that enter the mitochondria's electron transport chain (Figure 5) (11).

The reduced silicate mineral was observed to be a strong reducing agent by the direct reduction of cytochrome c. Cytochrome c is a crucial iron containing electron carrier protein in the respiratory electron transport chain of human cells. Cytochrome c is coupled to the production of ATP in the mitochondria where most ATP is created. ATP will participate in numerous critical reactions throughout the cell.

Energy in the form of ATP production by the three linked processes of glycolysis, the citric acid cycle (Kreb's cycle) and mitochondria respiration rely on the transfer of electrons from hydrogen and reduced hydrogen in these pathways by the important intermediates NAD, NADH, FAD, FADH, and FADH2. Numerous other metabolic processes in the cell depend on ATP and these hydrogen containing cofactors for fatty acid utilization, protein synthesis, DNA synthesis and repair, gene regulation, and deamination of amino acids for the excretion of ammonia in the form of urine, to name a few (11).

An antioxidant (reducing agent) is a compound that has an electron available to give to another molecule that needs one in order to stabilize its reactivity. Electrons orbiting around atoms prefer being paired or in even numbers (2, 4, 6, 8). Certain molecules, like the hydroxyl ion or superoxide ion, if not soon reduced (gain an electron) will have a strong enough affinity to attract electrons randomly from other molecules that will suffer structural and functional damage as this occurs. As the bio-molecules of cells are damaged in this way they begin to "age" or are not as normal as healthy cells.

The reduced silicate mineral supplement was also shown to scavenge the superoxide radical and the hydroxyl free radical. Hydroxyl radicals are among the most dangerous of oxygen free radicals that occur in biological systems and are the same as those formed by exposure to ionizing radiation. Both of these free radicals cause extensive oxidative damage to biological macromolecules such as DNA, membrane polyunsaturated fatty acid chains, and enzymes (12,13). Free radical damage (oxidative stress) is now considered to play a causative role in certain diseases such as Alzheimer's, rheumatoid arthritis, adult respiratory disease, cancer, heart and blood vessel diseases and aging (12,13).

Recent studies have shown that hydrogen in its reduced form scavenges active oxygen species and protects DNA from damage by oxygen free radicals (13). Reduced hydrogen is considered to be an ideal reactive species to counter active oxygen free radicals. Since water is the medium in which all organisms are born, composes the matrix of life and supports all molecular interactions, it is now understood that this is also part of its biological role (13,15). Various functions of water chemistry, its geological chemistry, even its genetically determined transport across the cell membrane and biochemical functions are now being investigated more extensively.

Hydrogen in its ground state holds one electron. It can also be found in nature to hold a second electron, known as the hydride (H: ^–) ion or reduced hydrogen. Hydrogen can be reduced (H:^–) such as when water is treated with electrolysis, or when other catalytic procedures are used (13-15). The hydrogen produced in the process is called active hydrogen because of its activated reducing potential as an antioxidant and it has a redox potential of –350 mV (13,15). Reduced water, the dissolved supplement, and Hunza water have reduction/oxidation potential values of –350 mV or lower indicating Iow dissolved oxygen and high dissolved hydrogen in solution which indicate the exclusive properties of reduced hydrogen and electron availability unique to these systems. Blended fresh fruit and vegetable juices also show negative redox values (16).

Other antioxidants (i.e. vitamin E, vitamin C, etc.) do not tend to display such negative electron availability, as measured by standard redox measurements, because of the various structures of the molecules, their individual chemical characteristics and functional proximity in reactions which tend to determine their antioxidant role. Ascorbic acid, vitamin C (+80 mV) has a relative redox potential much higher than NADH (–320 mV) or the reduced silicate supplement (–650 mV), therefore, an equal amount of ascorbic acid will not reduce the same amount of NAD+ at the same rate as Microhydrin (a stronger reducing agent) will.