A Silicate Mineral Supplement Microhydrin
®, Traps Reduced Hydrogen Providing In Vitro Biological Antioxidant PropertiesPatrick Flanagan MD. (MA), Flantech Corp. and Kimberly Purdy Lloyd MS.,Royal BodyCare, Inc.
Reprinted for educational purposes. Proceedings of the National Hydrogen Association, 10th Annual U.S. Hydrogen Meeting, Technology Advances Vol. 10, April 1999
Hunza Water
The Ultar Glacier of Hunza, West Pakistan has received interest in this century by both geologists and medical professionals as to the unique association towards unusual health and longevity of the people who consume the glacier stream water (4-7). Research has revealed that drinking the glacially pulverized rock flour of the Hunza river, eating a favorable diet, and community participation have been critical factors in achieving renowned longevity, a low rate of heart disease, and exceptionally good to excellent health in this community (4-7).
In
analyzing the correlation to health and the use of
glacial waters, it is apparent that not only do glacial
waters have an abundance of important trace minerals but
also the amorphous silicate derivatives themselves have
unique characteristics in their ability to structure
water and transport minerals and electrolytes (1,8,9).
Although the structural studies are complex as to the
kinetics of the silanol (SiOH) groups formed in different
silica (SiO
Silicates
Silicates comprise a mineral family and are among the most abundant components of the earth’s surface. Natural amorphous and crystallized silicates line rivers, glacier streams and marine environments. Weathering, pressure, ionic conditions and
temperatures through the ages produce a variety of soluble silicate derivatives. Numerous water sources including glacial rock suspensions have been analyzed for their silicate structures and mineral compositions (1,2). In particular, various mineral water sources and glacier regions have been linked to the health of the inhabitants who have depended on them for their drinking water (3,4). Some of the relationships between geochemical environment, health and disease are well documented such as iodine deficiency leading to diseases of the thyroid (i.e. goiter, hypothyroidism, cretinism, and increased risk of thyroid cancer) (3). Numerous minerals are now realized to per-form vital functions as cofactors and provide numerous structural and functional roles with-in cells and tissues. Recent evidence sug-gests that geochemical environments of water sources have a profound influence on the level of health in humans and animals (3-5).
Silicates tend to structurally arrange water molecules up to three layers (wetting). In the presence of molecular water, the silanol groups of small silicates ionize, producing mobile pro-tons that associate/dissociate with the surface to impart an electrical conductivity to the surface that attracts minerals and ions as shown in Figure 1 (1). These layers have been further described as the omega (o-innermost water layer), beta (- second water layer) and delta (d-outermost water layer) (Figure 1). The resulting surface-solution interface that exists at wetted mineral surfaces is called the electrical double layer or zeta potential. It is this characteristic that tends to transport small ions, minerals and electrolytes (i.e. hydrogen, iron, magnesium, calcium, and sodium, etc.) (1,8,9). These SiOH groups and the resulting water arrangements tend to cage or sieve minerals. They also hold hydrogen atoms within these structures (1,9,10). When hydrogen is further reduced a biological antioxidant property is maintained in the silicate mineral particle.
Methods
All capsules used in antioxidant and reduction analyses contained 250 mg of the naturally occurring food grade silica, potassium carbonate, magnesium sulfate and fatty acids developed by a proprietary process in a base of rice powder and were made available by Royal BodyCare, Inc. of Irving, Texas as its product Microhydrin.
Reduction Assays With Microhydrin
When the reduced hydrogen silicate mineral (Microhydrin) was dissolved or suspended in phosphate-buffered saline, pH 7.4, it directly reduced both cytochrome c and NAD + indicating its strong reducing capacities (antioxidant). When assayed in the standard assay for superoxide dismutase activity based upon the reduction of cytochrome c by xanthine oxidase/xanthine, it reduced cytochrome c. These overall reactions, the reduction of cytochrome c (Cyt c) and the reduction of NAD + by the reduced hydrogen silicate mineral (Mic) are shown below. NAD + reduction will be presented in detail later.
Mic(H- ) + Cyt c(Fe+3) Mic + Cyt c(Fe+2) + H
2Mic(H- ) + NAD+ 2Mic + NADH + H
Superoxide Scavenging Assays With Microhydrin
The
superoxide-mediated reduction of cytochrome c showed that
the reduced silicate mineral inhibited the
superoxide-mediated reduction of cytochrome c, indicating
that the reduced silicate is reducing, or scavenging, the
superoxide radical also:
Mic (H- ) + O.2- H + Mic + H2O2
In this assay Microhydrin was found to inhibit and prevent the reduction of cytochrome c by superoxide:
O.2- + Cyt c(Fe+3) O2 + Cyt c(Fe+2)
An alternative assay to further elucidate the activity of the reduced silicate mineral, observed the oxidation of epinephrine to adrenochrome by superoxide (unpublished data).
Microhydrin + O.2- + epinephrine H2O2 +adrenochrome
Superoxide free radical Hydrogen peroxide
Data is shown in Table 1 for superoxide dismutase-like (SOD) activity based on the oxidation of epinephrine by superoxide. In this assay the reduced hydrogen mineral silicate (Microhydrin) scavenged the superoxide ion again. Quantitatively, half-maximal inhibition was achieved at a Microhydrin concentration of 90 g/ml (Table 1).
When
concentrations of Microhydrin were at 60 g/ml or greater,
epinephrine oxidation was inhibited showing that the
silicate mineral reduced the superoxide free radical. The
assay conditions utilized superoxide dismutase at a
Microhydrin to reduce NAD+ to NADH
was also examined. The redox potential for this reaction
is 0.32 V ( 320 mV), requiring a fairly strong reducing
agent. Microhydrin reduced NAD+ to
NADH. The reduction of NAD+ was
verified by scanning the absorption spectrum from 200 nm
to 500 nm. In all subsequent studies NAD + reduction was
monitored as the difference between two wavelengths (340
nm to 500 nm).
The 500 nm wavelength served as a reference (independent of NAD + reduction, but control-ling for turbidity changes due to the light scat-tering effect of the silicate) (Figure 3). NAD + could be titrated by sequential addition of aliquots of Microhydrin in a linear fashion.
Microhydrin was suspended for 30 min. at 10mg/ml. Microhydrin was added incrementally to 60M NAD+ (Figure 3).
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 FeSO 4 which was
dissolved in distilled water. Fifty microliters of sample
solution, 0.18 M DMPO (spin trap,
5,5-dimethyl-pyrroline-N-oxide) (50 ml), 2mM H2O2
(50 ml) and 0.2mM FeSO4 (50
ml) 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 and 0.05 EPC-K1 mmol-equivalent/ mg and another
sample at 0.15 and 0.03 EPC-K1 mmol-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 cofac-tors participating in numerous bio-chemical 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 (Krebs 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 FADH 2 . 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).
Antioxidant Function of Microhydrin
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 dam-age 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 Alzheimers, rheumatoid arthritis, adult respiratory disease, cancer, heart and blood vessel diseases and aging
Hydrogen as an Antioxidant (electron donor)
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 sup-ports all molecular interactions, it is now understood that this is also part of its biologi-cal 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 low 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. Ascorbic acid however is required to act as an electron donor in specific enzymatic reactions that only will recognize it in order to function, as is the case with other antioxidant vitamins. Although many vitamins have shown antioxidant function in and beyond their role as enzymatic cofactors, antioxidants are now realized to act towards random free radicals generated by metabolism or detrimental intermediates and are pre-venting otherwise damaging free radical reactions
Since the reduced silicate can provide electrons for numerous reactions as an antioxidant it is now being further investigated as to these important antioxidant functions. Since biological molecules can be prevented from oxidation (loss of electrons) when reduced hydrogen is present to provide electrons the reduced silicate mineral is providing another function in addition to mineral and electrolyte transport. This antioxidant function may be an important factor in the silicate mineral system found in glacier waters renowned for longevity and enhanced health in communities that consume water with these properties (4-7).
Biological Function of Silica
Silicon (element name) has been discovered to be an important trace element necessary for cartilage and bone tissue formation (18). Silicates are essentially non-toxic when taken orally as evidenced by its use for over forty years as an over-the-counter ant-acid (magnesium trisilicate) for stomach indigestion. Silica compounds have also been used as pharmaceutical preparations (methylsilanetriol salicylate) for circulatory ischemias and osteoporosis (18). Silica compounds have unique buffering capacities shown often in their use in experimental biological systems (i.e. stabilizing biomolecules in column chromatography). A recent study found silica to provide a protective effect against high levels of aluminum in drinking water associated with cognitive impairment in the elderly especially when the pH was high (17). Solutions containing colloidal silicates have been used to successfully preserve canine hearts and kidneys for transplantation (19,21)
When the silicate mineral supplement was taken orally in an initial clinical study using human subjects it began showing an effect on urine pH. Most values showed changes to a more normal alkaline pH, considered beneficial towards health. A continuing state of blood and urine acidosis can indicate compromising health conditions. In this same study electrolyte and mineral transport were observed to optimize as seen by statistically significant (p >0.05) results in resistivity (1/conductivity) values in blood and urine in eight subjects after taking 4 capsules of the supplement per day for 18 days (20). Some nutritional investigators speculate that a silica mineral deficiency is involved in the causation of several human disorders including atherosclerosis, osteoarthritis, and hypertension, as well as, the aging process (18). Those speculations demonstrate the critical need for studying the importance of silica nutrition and function, especially in aging humans (18). Due to the unique chemical properties of silicate minerals, their antioxidant properties, long-term use, safety and recent research the scientific and nutritional communities continue to investigate the possibilities of these minerals as to their varied nutritional benefits.
References
1. Dove PM, Rimstidt JD. Silica-water interactions. In Heany PJ, Prewitt CT, Gibbs GV, (eds): Silica, Physical Behavior, Geochemistry, and Materials Application. Reviews in Minerology 29:259-301,1994.
2. Keller WD, Reesman AL. Glacial milks and their laboratory- simulated counterparts. Geol Soc Am Bull 74:61- 76,1963.
3. Hopps HC. Geochemical environment related to health and disease. Geological Society of America Special Paper 155:1-9,1975.
4. Keller WD, Feder GL. Chemical analysis of water used in Hunza, Pakistan. In Hemphill D.D. (ed): "Trace Substances in Environmental Health-XIII, Proceedings." University of Missouri-Columbia: 130-137, 1997.
5. Keller WD. Drinking Water: A geochemical factor in human health. Geological Society of American Bulletin 89:334-336, 1978.
6. Leaf A. Getting Old. Scientific American 229:44-52, 1973.
7. Murray MJ, Murray AB. Diet and Cardiovascular Disease in Centenarians of Hunza. Arteriosclerosis 4:546a, 1984.
8. Keller WD, Balgord WD, Reesman. Dissolved products of artificially pulverized silicate minerals and rocks: part I. Journal of Sedimentary Petrology 33:191-204,1963.
9. Keller WD. Argillation and direct bauxitization in terms of concentrations of hydrogen and metal cations at sur- face of hydrolyzing aluminum silicates. Bulletin of the American Association of Petroleum Geologists 42:233- 245,1958.
10. Sasamori R, Okaue Y, Isobe T, Matsuda Y. Stabilization of atomic hydrogen in both solution and crystal at room temperature. Science 265: 1691-1693, 1994.
11. Leninger AL, Nelson DL, Cox MM, (eds.): Principles of Bioenergetics. In "Principles of Biochemistry" 2nd ed. New York, Worth Publishers,1993.
12. Pryor WA, Shipley Godber S. Oxidative stress status: an introduction. Free Radical Biology and Medicine 0:173,1991.
13. Shirahata S, Kabayama S, Nakano M, Miura T, Kusumoto K, Gotoh M, Hayashi H, Otsubo K, Morisawa S, Katakura Y. Electrolyzed-reduced water scavenges active oxygen
species and protects DNA from oxidative damage. Biochemical and Biophysical Research Communications 234:269-274,1997.
14. Degani Y, Willner I. Photoinduced hydrogen evolution by a zwitterionic diquat electron acceptor. The functions of SiO 2 colloid in controlling the electron-transfer process. J. Am. Chem. Soc. 105: 6228-6233,1983.
15. Hayashi H. Water the chemistry of life, part IV. Explore 6, 1995: 28-31.
16. Howard C (ed). Microhydrin testing in various liquids. In "Microhydrin Technical Information" 3rd ed. Dallas: Arlington Publications 1-6,1998.
17. Jaqmin Godda H, Commenges D, Letenneur L, Dartigues JF. Silica and aluminum in drinking water and cognitive impairment in the elderly. Epidemiology 7:281- 285,1996.
18. Neilson FH. Ultratrace Minerals. In Shils ME, Young VR (eds): "Modern Nutrition in Health and Disease" 7th ed. Philadelphia: Lea and Febiger: 286-288, 1988.
19. Toledo-Pereyra LH, Sharp HL, Condie RM, Chee M, Lillehei RC, Najarian JS. Preservation of canine hearts after warm ischemia (zero to thirty minutes) and one to two days of hypothermic storage. A comparative analysis of crystalloid and colloid solutions with different osmolarity and ion composition. J Thorac Cardiovasc Surg 74:594-603,1977.
20. Smith L Jr., Purdy Lloyd K, Phelps K. Biological terrain assessment results of 14 subjects before and after testing with a supplement containing silicon bonded to reduced hydrogen ions. Journal of the American College of Nutrition 17, 522:1998.
21. Toledo-Pereyra LH, Condie RM, Simmons RL, Najarian JS. Complete protection of severely damaged kidneys by a silica gel plasma perfusate. Surg Forum 25,294-5:1974
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