Scavenging of Hydroxyl Free Radical with Microhydrin

When the reduced hydrogen silicate mineral supplement (Microhydrin) was analyzed for antioxidant activity by Electron Spin Resonance (ESR) techniques, the reduced silicate mineral supplement showed activity toward the scavenging of hydroxyl free radical (OH) at a concentration of 0.625 mg/ml (Figure 4). Hydroxyl radicals were generated by the Fenton reaction. All reagents were dissolved in 0.1M potassium phosphate buffer, pH 7.4, with the exception of FeSO4 which was dissolved in distilled water. Fifty microliters of sample solution, 0.18 M DMPO (spin trap, 5,5-dimethyl-pyrroline-N-oxide) (50 µl), 2mM H2O2 (50 µl) and 0.2mM FeSO4 (50 µl) were mixed for 10 seconds and quickly transferred into a quartz flat cell (200 ml capacity, JEOL, Tokyo, Japan). Exactly 30 seconds after the addition of FeSO4, ESR spectra of the DMPO-OH spin adducts were recorded (unpublished data) (figure 4).

Two samples of the supplement were evaluated one showing hydroxyl free radical scavenging at 0.19 ± 0.05 EPC-K1 µmol-equivalent/mg and another sample at 0.15 ± 0.03 EPC-K1 µmol-equivalent/mg (unpublished data).

Conclusion
Reduced Hydrogen In The Energy Cycle

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-phosphate 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).