CN113087108A

CN113087108A - Nano hydrogen free radical water for targeted eliminating cytotoxic active oxygen and preparation thereof

Info

Publication number: CN113087108A
Application number: CN201911334184.4A
Authority: CN
Inventors: 福田浩平; 解冰
Original assignee: Individual
Current assignee: Individual
Priority date: 2019-12-23
Filing date: 2019-12-23
Publication date: 2021-07-09

Abstract

The invention discloses nano hydrogen free radical water for removing cytotoxic active oxygen in a targeting manner and a preparation method thereof, wherein the nano hydrogen free radical water comprises hydrogen bubbles below 200nm and hydrogen free radicals wrapped by the hydrogen bubbles; the preparation comprises the following steps: discharging hydrogen into water through a gas supply device and an air explosion plate, and performing ionization treatment to obtain hydrogen radicals wrapped by hydrogen bubbles below 200 nm; and guiding out the liquid, filling and sealing to obtain the nano hydrogen free radical water. And the ratio of the amount of hydrogen discharged by each micropore of the gas explosion plate during ionization treatment to the vibration frequency during ionization is controlled to be less than or equal to 300. The method not only realizes the production of hydrogen bubbles below 200 nanometers, but also successfully realizes the wrapping of hydrogen free radicals by the nano hydrogen bubbles, and obtains a new minimum strongest antioxidant of antitoxic active oxygen free radicals.

Description

Nano hydrogen free radical water for targeted eliminating cytotoxic active oxygen and preparation thereof

Technical Field

The invention belongs to the field of preparation of nano hydrogen free radical water, and particularly relates to a preparation method of nano hydrogen free radical water for removing cytotoxic active oxygen in a targeted manner.

Background

Cancer has become the most major threat to human life health. According to the statistics of the international cancer research center of the world health organization, more than 1000 new cancer cases are newly added every day in the world, and the number of new cancer patients reaches 1500 ten thousand every year in the world in 2020, thus showing a runaway trend.

The latest national cancer statistical data released by the national cancer center in 2019 and 1 month show that the number of new malignant tumor cases in China in 2015 is about 392.9 ten thousands, and the new malignant tumor cases are increased by about 4% in recent decades, wherein the lethality of lung cancer (17.8%), gastric cancer (10.4%) and liver cancer (8.8%) is the greatest. Cancer not only afflicts the body of a patient, but also brings heavy economic burden to families, and further brings immeasurable loss to the country and the society.

Medical studies have demonstrated that more than 85% of cancers are a by-product of human metabolism (cellular variation) rather than genetic. The mechanism of free radicals is the basis of all cancer mechanisms. The healthy metabolism process of human body is in a state of free radical with a proper level, namely, a certain amount of free radical can ensure to remove garbage in vivo, maintain the oxidation resistance of the organism, promote the stress reaction of cells to generate the immunity of the organism and the like. However, due to the over-development of modern industrial society, the living environment of human beings is flooded with countless free radicals, and the human beings are exposed to the surrounding and attacking of the free radicals all the time. Excessive diet and poor lifestyle can cause disruption of normal cell metabolic processes, stimulate abnormal activity in some cells, and produce more Reactive Oxygen Species (ROS) than normal cells. Thus, the potential of free radicals from the inside and outside of the body to attack the human body is accumulated, and the excessive runaway of the free radicals in the human body is caused.

The research shows that: the behavior of Reactive Oxygen Species (ROS) is associated with the promotion of cell proliferation, DNA synthesis, survival, cell migration, invasion, tumor metastasis and angiogenesis. The principle of the method is that oxygen is a respiratory state element necessary for normal metabolism of cells, the oxygen respiratory state changes, the cell metabolism is in a stress reaction state, and mechanism changes can be caused after the cell metabolism is long-term, and the method is mainly divided into two parts: on one hand, free radicals attack cell membranes of normal cells or damage cell nucleuses, and compete with genes for electrons, so that the normal respiratory state of the cells is changed, the oxygen regulatory gene sequence mechanism in the cell nucleuses is damaged, and the gene mutation of the cells is caused; on the other hand, the genetically altered cells can attack other normal cells through toxic active oxygen (such as hydroxyl radical) carried by the cells, and similar to infectious diseases, cell variation is induced in a large area, and the purpose of rapid proliferation is achieved. Moreover, the attack route initiated by the cells carrying toxic active oxygen is uncertain, causing various damages to the organism and generating various difficult and complicated diseases.

While free radicals and derivatives thereof in the human body often act in the form of hydroxyl radicals (e.g., phenols), at the same time, the excess hydroxyl radical OH is also a major "weapon" for the altered cells to attack normal cells, and is therefore referred to as "toxic reactive oxygen species" in modern medicine. Therefore, modern medicine has been working on the therapeutic approach of achieving inhibition of tumor cell proliferation by killing cancer-causing cells over large areas (chemotherapy), and by the interaction of antioxidants with reactive oxygen species.

At present, the antioxidant products in the medical world and the market are basically inclined to synthesize chemical preparations to attack the suspected targets in vivo in a large area, even though the products are 'attacked by poison'. This has the consequence that chemically synthesized preparations do not ensure a reaction with toxic reactive oxygen radicals, on the one hand, do not eliminate toxic activity on tumor cells or cancer cell bodies (ineffective reactions), and on the other hand, inappropriate chemical reactions can lead to unknown side effects (toxic side effects). This is common in the clinic.

It is well known that hydrogen is the most basic antioxidant substance for all life on earth, and it is probably the most superior antioxidant for the various life forms so far, because the hydrogen radical H and OH produce the most appropriate chemical reaction: h +. OH ═ H₂And O. However, hydrogen radicals (. H) are very active mediators which are difficult to exist alone in nature, and if there are two, they automatically combine into hydrogen H immediately₂And release energy. Therefore, it is difficult to find H alone in nature.

Over the last decade, much development has been explored around the function of hydrogen. Different types of research and development test products are currently on the market, among which products produced by the electrolytic water process, which is the first generation technology represented by china, and products produced by the gas permeation membrane process, which is the second generation technology represented by part of the japanese enterprises.

However, with the electrolytic method, i.e., the method of providing positive and negative electrodes in water to decompose water into hydrogen and oxygen, thereby generating hydrogen water (bubbles), the disadvantages are obvious:

1. most of common bubbles discharged into water can not be dissolved in water, can quickly rise and burst on the surface of the liquid to be emitted into the atmosphere; even if dissolved for a long time, the hydrogen concentration at normal temperature and pressure cannot reach a supersaturated state of 1.58ppm or more.

2. There is biological safety: (1) if water with more ions is electrolyzed, harmful substances such as ozone, chloroform and the like can be generated at the same time; (2) the metal layer of the heavy metal component such as platinum on the surface of the electrolytic film may fall off, causing additional physical damage.

The gas permeation membrane method solves the disadvantages of the first generation technology (electrolysis method) by injecting gas into a liquid solvent under a pressure state exceeding a certain pressure by means of a gas permeation membrane having a specific gas permeation performance condition, but the method is limited by the material of the hollow fiber membrane, so that only hydrogen bubbles above 500nm can be produced, the solubility is limited to a certain degree, and the stability of the bubbles needs to be further improved. Meanwhile, the product has the problem of biological safety of unknown substances precipitated in water. These are all to be solved by the new technical invention.

The invention mainly aims to solve the defects of the first generation technology and the second generation technology, target elimination is carried out aiming at the basis of a cancer mechanism, namely excessive toxic free radicals, and a 'brake mechanism' for inhibiting the proliferation and the diffusion of cancer cells is created, thereby creating a prerequisite for the next treatment and cell repair.

Disclosure of Invention

In order to overcome the problems in the prior art, the invention realizes important technical change by adopting a physical hydrogen gas-liquid mixing method, perfectly solves all the defects of an electrolytic method, and improves the production limit of a gas permeation membrane method to a brand new level.

The method of the invention not only realizes the production of hydrogen bubbles below 200nm (optimally 100nm, the smallest diameter which can be detected so far), but also successfully produces the nanometer hydrogen radical (H) and the nanometer hydrogen (H) which are the strongest antioxidants with new anti-toxicity active oxygen radicals₂) Bubble and successfully realize the encapsulation of hydrogen radicals by nano hydrogen bubbles.

One of the purposes of the invention is to provide a preparation method of nano hydrogen free radical water, which comprises the following steps:

step 1, discharging hydrogen into water through a gas supply device and an air explosion plate, and performing ionization treatment in the water to generate hydrogen radicals wrapped by hydrogen bubbles below 200 nm;

and 2, guiding out the liquid, filling and sealing to obtain the nano hydrogen free radical water.

In a preferred embodiment, the hydrogen concentration in step 1 is above 99%, preferably above 99.8%. The medical hydrogen can be purchased in the market and can also be purchased by equipment.

In a preferred embodiment, in step 1, the aeration plate is disposed in water and connected to the gas supply device.

In a further preferred embodiment, the hydrogen is discharged by means of a gas supply onto a gassing plate placed in the water.

In a preferred embodiment, the aeration plate is made of a nano-active carbon fiber material.

In a more preferred embodiment, the carbon nanofibers have micropores with a pore diameter of 1.5 μm or less, preferably 0.01 to 1.5 μm, and more preferably 0.1 to 1.0. mu.m.

The aeration plate is prepared from a nano activated carbon fiber material disclosed in the prior art, the nano activated carbon fiber is distributed with micropores with irregular paths, so that a plurality of air supply passages extending in different directions are formed in the carbon fiber, the surface of the carbon fiber is uniformly distributed with micropores in a flaky shape, the number of the micropores is not particularly limited (the number of 200-800 ten thousand pores is preferably 300-500 ten thousand), the aeration plate is determined according to equipment and process, the more the aeration plate is, the better the introduction amount of the gas introduced into water is theoretically, and the higher the introduction amount of the gas is.

In a preferred embodiment, in step 1, the ionization treatment is carried out by means of vibrational radiation.

Wherein, the nano-scale hydrogen bubbles below 200nm can be successfully obtained by the dual functions of high-frequency high-energy vibration radiation and an air explosion plate, meanwhile, part of the hydrogen bubbles are shaken (pulverized) into hydrogen free radicals under the high-frequency high-energy condition to realize ionization, and the hydrogen free radicals are successfully wrapped by the nano-scale hydrogen bubbles to form H₃(Hydrogen radical. H and Hydrogen gas H)₂In combination) ensures that the nano-hydrogen radicals are stably dissolved in water.

In a further preferred embodiment, the conditions of the vibrational radiation are: the vibration frequency is above 30000Hz, and the amplitude is below 2 mm.

In a further preferred embodiment, the conditions of the vibrational radiation are: the vibration frequency is 50000Hz or above, and the amplitude is 1mm or below.

In a preferred embodiment, a vibrator is arranged below the gas explosion plate for vibration radiation.

In a further preferred embodiment, the vibrator vibrates in a range of an angle of-15 degrees to 15 degrees with respect to an exhaust surface of the explosion plate, and the angle formed by the vibrator and the exhaust surface is preferably 0 degrees, that is: the vibrator applies vibration in the direction perpendicular to the gas discharge direction to realize ionization of the hydrogen bubbles.

In a preferred embodiment, the gas discharge amount [ μm ] of each micro-hole on the gas blast plate is controlled³Minute/min]) /(vibration frequency [ Hz ]])≦300。

In a further preferred embodiment, the gas discharge amount [ μm ] of each micro-hole on the gas blast plate is controlled³Minute/min]) /(vibration frequency [ Hz ]]) 200 or less, more preferably 100 or less.

In step 1, the discharge rate of hydrogen is satisfied (discharge amount of gas [ mu m ] per minute hole on the gas explosion plate)³Minute/min]) /(frequency of vibration of vibrator [ Hz [)]) At ≦ 300, the amount of gas discharged per cell is controlled by a control valve. The reason for setting this condition is that the inventors have found through a lot of experiments that the smaller the set value, the easier the nanobubbles are generated during the ionization in step 1, and the smaller the diameter of the nanobubbles (desirably 200nm or less, more desirably 100nm or less), the easier the nanobubbles are dissolved in water, and the better the stability.

In the invention, the nano hydrogen free radical water obtained in the step 2 needs to be placed in a shade place, and high temperature, illumination and violent shaking are avoided.

The second object of the present invention is to provide nano hydrogen radical water obtained by the production method of the first object of the present invention, in which hydrogen radicals coated with hydrogen bubbles are dispersed.

The functional components (hydrogen bubbles and hydrogen radicals wrapped by the hydrogen bubbles) in the nano hydrogen radical water are only one part per billion of common water molecules due to the fact that the diameter of the functional components is smaller than 200nm, particularly the hydrogen radicals are ultra-small in size and only 10-15 ten-thousandths of the average size of human cells, and the nano hydrogen radical water has extremely high permeability. After being drunk by a human body, the medicine can be quickly diffused into blood, living tissues and cell tissues, and particularly can easily pass through blood brain barriers and other parts which cannot be reached by common medicines or nutrients or enter any cell through blocked blood vessels, so that the full coverage of target search is realized.

Wherein, after entering human body, the nano hydrogen bubble reaches the same air pressure environment as human body in a short time, the hydrogen bubble starts to release hydrogen free radical with extremely strong activity, and the hydrogen free radical H is reduced into hydrogen free radical, the hydrogen free radical H can quickly and automatically search cells (tumor or cancer variant) of hydroxyl free radical (OH) in blood tissue, and the cells react to produce water (H +. OH) according to the activity degree thereof₂O), targeted and harmless elimination of toxic active oxygen (hydroxyl radical. OH) in vivo, timely elimination of toxic activity on cell body, inhibition of tumor cell proliferation, and blocking of tumor cell carried with the toxic active oxygenThe spreading and metastasis of toxic and active cells, and the 'braking' of cancer.

Experiments prove that the high activity of the hydrogen free radical and the hydrogen gas not only can consume toxic and harmful active oxygen free radicals accumulated in the organism and reduce oxidative stress, but also can stimulate the absorption and metabolism of various nutrient substances, promote the energy conversion, improve metabolic disorders such as diabetes, hyperlipidemia, visceral fat obesity and the like and improve the absorption and utilization of nutrient substances needed by the body, and found cases prove that the hydrogen free radical water can prevent more than 38 diseases such as arteriosclerosis, cognitive decline, lung injury, neuropathic pain and the like caused by the stress in the body. The medical application can be further expanded in the future.

The other purpose of the invention is to provide a device for preparing nano hydrogen free radical water, which is preferably used for carrying out one of the purposes of the invention, wherein the device comprises a base container, an explosion plate, an ionization device and a gas supply device, wherein water is filled in the base container, the explosion plate and the ionization device are both arranged in the water, and the explosion plate is connected with the gas supply device.

In a preferred embodiment, the aeration plate is made of a nano activated carbon fiber material, and micropores are formed in the nano activated carbon fiber material, wherein the pore diameter of the micropores is less than 1.5 μm.

In a further preferred embodiment, the pore size of the micropores is from 0.01 μm to 1.5. mu.m, preferably from 0.1 μm to 1.0. mu.m.

In a preferred embodiment, the gas explosion plate is connected to the gas supply device, so that hydrogen bubbles are generated in water after hydrogen passes through the gas explosion plate.

In a preferred embodiment, the ionization device is disposed below the squib plate, and preferably, the ionization device includes a vibrator.

In a further preferred embodiment, the vibrator is vibrated at an angle ranging from-15 degrees to 15 degrees with respect to the exhaust surface of the explosion plate, and the angle formed by the vibrator and the exhaust surface is preferably 0 degrees, that is: the vibrator applies vibration in the direction perpendicular to the gas discharge direction to realize ionization of the hydrogen bubbles.

The ionizing device used in the embodiment of the present invention is a high-frequency high-voltage vibration radiation device (model: UFB-N4B) provided by guang jiu corporation of japan, but the ionizing device specifically used for carrying out the method of the present invention is not limited to the above-mentioned high-frequency high-voltage vibration radiation device, and may be any high-frequency high-voltage vibration radiation device disclosed in the prior art as long as the above-mentioned vibration conditions (vibration frequency of 30000Hz or more and amplitude of 2mm or less, preferably, vibration radiation conditions of 50000Hz or more and 1mm or less) can be achieved.

Compared with the prior art, the invention has the following beneficial effects:

(1) high biological safety. The benefits of hydrogen are being explored. However, the mechanism of action of hydrogen gas inhaled by the human body is unclear, and the patient is at risk of other side effects, thus limiting its medical use. Compared with other technologies, the functional component of the hydrogen radical water is hydrogen radicals wrapped by hydrogen bubbles, the hydrogen radicals are reduced and released in a human body, the activity of the hydrogen radicals is utilized to perform Brownian movement, and after the hydrogen radicals are combined with active oxygen radicals (hydroxyl radicals) which are not necessary for the human body, the metabolism function of the human body is improved, and finally, the metabolites are generated water (& H & lt + & gt & gtOH & gtH & lt + & gt & ltH & gt)₂O), 100% without toxic and side effect. It can be used for neonate, pregnant woman, and patients with various diseases. This was also confirmed by animal toxicity testing.

(2) High concentration. The hydrogen radical is a very active medium, and even if H is prepared instantly, H can be combined quickly into H if not wrapped and isolated in time₂Therefore, H is not present at normal temperature and pressure. And hydrogen (H)₂) The second generation technology gas permeation membrane method is disclosed that the highest content of producible hydrogen bubbles is 8 million fine bubbles of 500 nanometers per 1 milliliter. In the hydrogen radical water, 1.5 hundred million hydrogen bubbles below 200 nanometers are contained in 1 milliliter of water, and the product has the highest solubility at present.

(3) High stability. Due to poor stability of hydrogen, prolonged storage time, and extrusionOr in the shaking process, the phenomenon of hydrogen overflow exists. After the hydrogen gas (including the hydrogen absorption mode publicized in the market at present) overflows from the hydrogen gas, most of the hydrogen gas (hydrogen H) is in a gaseous state after being drunk (absorbed) into the human body₂) The form (oral and anal) escapes and the application function is lost.

The method successfully realizes the long-term stability of the nano hydrogen bubbles, and further ensures the stable existence of the hydrogen free radicals in the valid period (the half-life period of the hydrogen is 3 months).

(4) High permeability. The functional components (hydrogen bubbles and hydrogen radicals wrapped by the hydrogen bubbles) in the nano hydrogen radical water are only one part per billion of common water molecules due to the fact that the diameter of the functional components is smaller than 200nm, particularly the hydrogen radicals are ultra-small in size and only 10-15 ten-thousandths of the average size of human cells, and the nano hydrogen radical water has extremely high permeability. After being drunk by a human body, the health-care wine can quickly spread into blood, living tissues and cell tissues, and particularly can easily pass through blood brain barriers and other parts which cannot be reached by common medicines or nutrients or enter any cell through a blocked blood vessel, so that full coverage for target search is realized;

(5) high effectiveness. The method of the invention not only realizes the production of hydrogen bubbles below 200nm (optimally 100nm, the smallest diameter which can be detected so far), but also successfully produces the nanometer hydrogen radical (H) and the nanometer hydrogen (H) which are the strongest antioxidants with new anti-toxicity active oxygen radicals₂) Soaking, and successfully realizing the wrapping of hydrogen free radicals by nano hydrogen bubbles to obtain a new minimum strongest antioxidant of antitoxic active oxygen free radicals; in application, the health-care food can prevent more than 38 diseases such as arteriosclerosis, cognitive decline, lung injury, neuropathic pain and the like caused by internal stress, can stimulate the absorption and metabolism of various nutrient substances, promotes energy conversion, improves metabolic disorders such as diabetes, hyperlipidemia, visceral fat obesity and the like, and improves the absorption and utilization of nutrient substances required by the body. The medical application can be further expanded in the future.

However, it should be noted that: due to the neutralization effect of the nano hydrogen free radical bubbles, the functions of the nano hydrogen free radical bubbles are not 'treatment', but 'toxic active oxygen elimination', 'tumor cell proliferation inhibition', 'cancer cell movement depending on activity' and the like, and the functions of prevention, health care, recovery and the like are not 'treatment', and the nano hydrogen free radical bubbles can not replace the treatment of patients. It is particularly noted that the time interval is taken into consideration when administering the phenolic formulation (containing hydroxyl groups) in order to reduce its therapeutic effect.

Drawings

FIG. 1 shows a front cross-sectional view of an apparatus according to example 1 of the present invention;

FIG. 2 shows a side cross-sectional view of the device according to example 1 of the present invention;

FIG. 3 shows the imaging results of the hydrogen radical micro-camera instrument obtained in Experimental example 2;

FIG. 4 shows the Brownian motion results of hydrogen bubbles in water obtained in Experimental example 3;

fig. 5 shows the comparison results of the surface tension (permeability) of the normal tap water in experimental example 4, the nano hydrogen radical water obtained in comparative example 1, comparative example 2 and example 1;

fig. 6 shows soil permeability results of nano hydrogen radical water obtained in example 1 and comparative example 2 of experimental example 4;

FIG. 7 is a graph showing the results of the measurement of the active oxygen scavenging ability of cells using the nano hydrogen-based water of comparative example 1 and example 1 in Experimental example 6;

FIG. 8 is a SEM result graph showing degradation of cell morphology by nano-hydrogen radical water obtained in example 1 of Experimental example 7;

FIG. 9 shows the restoration of cell growth after the removal of active oxygen in Experimental example 8;

FIG. 10-1 shows the results of a computed tomography scan of the intact lung of a healthy rabbit in Experimental example 10;

fig. 10-2 shows the computed tomography results of pleural viscera after a blank group of lung metastases on day 14 in experimental example 10;

FIG. 10-3 shows the computed tomography results of lung metastasis in the anticancer treatment group at day 32 in Experimental example 10;

fig. 10-4 show the computed tomography results of lung metastases for the example 1 (nano-hydro radical water set) group on day 90.

The reference numerals in FIGS. 1-2 illustrate: 1-a hydrogen delivery pipeline; 2-a control valve; 3-a base container; 4-blasting gas plate; 5-an ionization device.

Detailed Description

While the present invention will be described in detail with reference to the following examples, it should be understood that the following examples are illustrative of the present invention and are not to be construed as limiting the scope of the present invention.

In an embodiment, the ionization apparatus and the gas stick used are provided by garland corporation of japan. Specifically, in the embodiment, the gas explosion plate is made of a nano activated carbon fiber material, and the nano activated carbon fiber is distributed with micropores with irregular paths, so that a plurality of gas supply passages extending to different directions are formed inside the carbon fiber, and the micropores are uniformly distributed on the surface in a sheet shape.

[ example 1 ]

Introducing 4L purified water into purified water tank at 20 deg.C, starting operation, regulating air flow control valve, passing through air explosion plate with 300 ten thousand micropores with average pore diameter of 1 μm, and making the air explosion plate have a diameter of 4000mm³Hydrogen gas with a concentration of 99.7% or more by volume is discharged at a rate of/min (discharge amount of hydrogen gas discharged from the micro-holes of the gas explosion plate is μm)³Under the condition that the frequency/minute)/(the vibration frequency Hz of the vibrator) is 33, vibration radiation with the frequency of 30000Hz and the amplitude of 1mm is continuously applied to an ionization device for 2 minutes to obtain the nano hydrogen radical water.

Wherein the discharge amount (mum) of hydrogen discharged from the micropores of the gas explosion plate³Min) is controlled by a gauge panel controlling a valve (reference 2 in fig. 1 and 2). The ionization device comprises a vibrator, and the angle formed by the vibrator and the exhaust surface of the gas explosion plate is 0 degree.

The obtained nano hydrogen free radical water has obvious Tyndall phenomenon.

The tyndall phenomenon: because the particle radius of the hydrogen bubbles is about 100nm and is smaller than the wavelength of visible light (400 nm-700 nm), when the visible light penetrates through the hydrogen free radical water, the visible light can generate obvious scattering effect, namely the Tyndall phenomenon is generated.

[ example 2 ]

Introducing 4L purified water into purified water tank at 20 deg.C, starting operation, regulating air flow control valve, passing through gas explosion plate with 300 ten thousand micropores with average pore diameter of 1.5 μm, and making the gas explosion plate have a diameter of 4000mm³Hydrogen gas with a concentration of 99.7% or more by volume is discharged at a rate of/min (discharge amount of hydrogen gas discharged from the micro-holes of the gas explosion plate is μm)³Under the condition that the frequency/minute)/(the vibration frequency Hz of the vibrator) is 100, vibration radiation with the frequency of 50000Hz and the amplitude of 2mm is continuously applied to an ionization device for 2 minutes to obtain the nano hydrogen radical water.

Among them, the discharge amount of hydrogen gas (μm)³Min) is controlled by controlling the valve gauge panel. The ionization device comprises a vibrator, and the vibrator applies vibration in an angle range of-15 degrees to 15 degrees with the exhaust surface of the gas explosion plate.

The obtained nano hydrogen free radical water has obvious Tyndall phenomenon.

Comparative example 1

The procedure of example 1 was repeated except that: the hydrogen is directly injected into the water storage device from the hydrogen pipeline without adopting an explosion plate, and the vibrator is directly started to carry out ionization treatment.

Comparative example 2

Hydrogen gas produced by a gas permeable membrane according to the patent document (japanese patent No. 5746411) was produced by market as hydrogen bubble plain water of 500nm, product number: cool and refreshing drink water (manufactured by Ivy Japan K.K., production date: 2019-10-10).

Comparative example 3

The procedure of example 1 was repeated except that: the gas explosion plate used contained 200 ten thousand parallel arranged hole shapes with an average pore diameter of 5 μm.

Comparative example 4

The procedure of example 1 was repeated except that: control (blast plate micropore discharge hydrogen discharge amount mum)³The/min)/(oscillation frequency Hz of the oscillator) is 500.

Comparative example 5

The procedure of example 1 was repeated except that: the ionizing device was continuously subjected to vibrating radiation having a frequency of 10000Hz and an amplitude of 5mm for 2 minutes.

[ Experimental example 1 ]

The products obtained in example 1 and comparative examples 1 to 5 were compared using a Malvern (Malvern) nanoparticle tracker in the united kingdom, and the results are shown in table 1 below:

table 1:

case(s)

Tyndall phenomenon

Average diameter of hydrogen bubble

Hydrogen bubble content

Half life

Osmotic power

Example 1

Is obvious

100nm

1.5 hundred million/mL

3 months old

High strength

Comparative example 1

Is free of

Not detected

Is free of

0

Weak (weak)

Comparative example 2

In general

500nm

4 million/mL

1 month

Is stronger

Comparative example 3

Is obvious

200nm

8 million/mL

3 months old

High strength

Comparative example 4

Is more obvious

230nm

7.8 million/mL

3 months old

High strength

Comparative example 5

Is more obvious

210nm

7.5 million/mL

3 months old

High strength

[ Experimental example 2 ] laser-induced fluorescence imaging method for detecting ionized free radicals

For the product obtained in example 1, the charging of visible ions (hydrogen radicals) was clearly imaged by a miniature camera instrument by charge measurement before and during bubble wrapping (i.e. tracking the bubble wrapping process), except for the display value measured by an ammeter, and the result is shown in fig. 3.

[ Experimental example 3 ]

The product obtained in example 1 was examined by a marvens nanoparticle tracker, and it was clearly found that the nano-hydrogen bubbles make irregular brownian motion in water, and the results are shown in fig. 4, in which (a) in fig. 4 shows brownian motion of each hydrogen bubble and (b) in fig. 4 shows the motion trajectory of the hydrogen bubble.

[ Experimental example 4 ] measurement of osmotic Capacity

(1) Surface tension is a key index determining the permeability of water quality, and the molecular diameter of water quality is a determining factor determining surface tension (permeability). Fig. 5 is a comparison of surface tension (permeability) of the nano hydrogen radical water obtained from the common tap water, comparative example 1, comparative example 2 and example 1.

(2) The soil permeability of the nano hydrogen radical water obtained in example 1 and comparative example 2 was measured, and the results are shown in fig. 6 and table 2.

Table 2:

[ Experimental example 5 ] toxicity test

The product obtained in example 1 was subjected to 2-week repeated drug toxicity test using rats and glycyrrhiza monkeys based on ICH international guidelines, and showed no abnormal reaction, weight gain, and total survival, and no significant acute toxicity was observed compared with the control group, confirming a good safety effect. The results of the specific toxicity tests are shown in table 3.

Table 3: results of acute toxicity test

And (4) t test: p >0.05 compared to the blank control. Note: "-" is normal

[ Experimental example 6 ] measurement of cell active oxygen targeting scavenging ability

The cell active oxygen scavenging capacity of the nano hydrogen radical water obtained in example 1 on human skin culture medium was detected by redox indicator CDCF fluorescence (see fig. 7).

(1) Fluorescence showing the presence of a large amount of active oxygen can be seen in the nucleus of human skin cells treated without the product obtained in example 1 (without using nano hydrogen radical water) [ fig. 7 (a) ]; in human skin cells treated with nanobubble hydrogen medium, fluorescence indicating reactive oxygen species near the nucleus and cytoplasm are scavenged [ fig. 7 (b) ];

(2) comparative example 1 was directly subjected to ionization treatment, and found that the hydrogen radical retention therein was significantly reduced, and fluorescence showing the presence of a large amount of active oxygen was observed in the nuclei of human skin cells treated therewith.

[ Experimental example 7 ] comparison of the ability to kill Ehrlich ascites tumor cells

Observing the anti-tumor effect of the nano hydrogen free radical water obtained in the example 1 on Ehrlich Ascites Tumor (EAT) cells by using a crystal violet staining method; the nano hydrogen radical water obtained in example 1 was observed by a Scanning Electron Microscope (SEM) to deteriorate the cell morphology to evaluate the cancer suppressing effect, and the result is shown in fig. 8.

FIG. 8 shows the morphological changes of Ehrlich ascites tumor cells treated with bubble water, in which the nano-hydrogen radical water obtained in example 1 was used in the experimental group and the nano-hydrogen radical water was not used in the control group. In fig. 8, it can be seen that after the nano hydrogen radical bubble water treatment of the invention in example 1, the cilia on the surface of the tumor cells are reduced, the cells are smooth, and even the cells are cracked.

[ Experimental example 8 ] measurement of cell restoration ability after removal of active oxygen

After scavenging of reactive oxygen species from the cells in the human skin medium of Experimental example 6, the cells were then observed and measured for restoration of growth, as shown in FIG. 9, in which (a) is the cells after elimination of the reactive oxygen species and (b) is the cells measured for restoration of growth after one week.

[ Experimental example 9 ] Slow Release test for adverse stress on dialysis

The nano-hydrogen bubble water prepared in example 1 was provided to 250 dialysis patients (divided into two groups) participating in a 12-month clinical trial, in which 125 patients each took the product of example 1 before or during dialysis, and the other 125 patients did not. The two groups were exchanged in the next month. The results show that the nano hydrogen bubble water prepared in the example 1 can obviously relieve the side effects of fatigue, pruritus, hypertension and the like after dialysis. The results are shown in Table 4, in which M1 to M12 represent the first to twelfth months, respectively.

Table 4:

[ Experimental example 10 ] antitumor Effect against Rabbit squamous cell carcinoma (prevention of metastasis of cancer cells)

Healthy adult white rabbits were cultured by local (ear) injection of Head and Neck Squamous Cell Carcinoma (HNSCC), a cancer that frequently metastasizes in humans and animals and shows a high mortality rate, and were subjected to comparative treatments in three groups (14 per group) of a blank control group (no treatment), anti-cancer drug treatment, intraperitoneal injection (instead of oral administration) of experimental example 1, and the like, respectively, and the anti-tumor (cancer cell metastasis-preventing) effect was observed.

The lung cancer cell metastasis of the control group was scanned using Computed Tomography (CT), confirming that no lung metastasis occurred in the patient after the nano hydrogen radical aqueous solution of example 1 was used. As shown in particular in fig. 10-1 to 10-4. In which fig. 10-1 shows the intact lung of a healthy rabbit, fig. 10-2 shows the pleural viscera after lung metastasis in the blank group on day 14, fig. 10-3 shows lung metastasis in the anticancer-treated group on day 32, and fig. 10-4 shows lung metastasis in the example 1 group on day 90 (lung metastasis did not occur).

Claims

1. A method for preparing nano hydrogen free radical water comprises the following steps:

2. The production method according to claim 1, wherein, in step 1,

the gas explosion plate is arranged in water and connected with the gas supply device, and preferably, hydrogen is discharged to the gas explosion plate arranged in the water through the gas supply device.

3. The method according to claim 2, wherein the gas explosion plate is made of a nano activated carbon fiber material, and the nano activated carbon fiber material has micropores with a pore diameter of 1.5 μm or less, preferably 0.01 to 1.5 μm, and more preferably 0.1 to 1.0 μm.

4. The production method according to claim 1, wherein in step 1, the ionization treatment is performed by means of vibrational radiation; preferably, a vibrator is arranged below the gas explosion plate for vibration radiation.

5. The manufacturing method according to claim 4, wherein in step 1, the vibrator vibrates within an angle range of-15 degrees to 15 degrees with respect to the exhaust surface of the gas explosion plate during vibration irradiation, and preferably, the angle formed by the vibrator and the exhaust surface of the gas explosion plate is 0 degree.

6. The production method according to claim 1, wherein in step 1, the conditions of the vibrational radiation are: the vibration frequency is above 30000Hz, and the amplitude is below 2 mm; preferably, the conditions of the vibrational radiation are: the vibration frequency is 50000Hz or above, and the amplitude is 1mm or below.

7. The production method according to any one of claims 1 to 6,

control (gas discharge amount [ mu m ] of each micro-hole on gas explosion plate³Minute/min]) /(vibration frequency [ Hz ]]) ≦ 300, preferably 200 or less, more preferably 100 or less.

8. Nano hydrogen radical water obtained by the preparation method according to any one of claims 1 to 7.

9. A device for preparing nano hydrogen radical water, preferably used for carrying out the preparation method according to any one of claims 1 to 7, wherein the device comprises a base container, an explosion plate, an ionization device and a gas supply device, wherein water is filled in the base container, the explosion plate and the ionization device are both arranged in the water, the explosion plate is connected with the gas supply device, and the ionization device is arranged below the explosion plate.

10. The apparatus of claim 9,

the ionization device comprises a vibrator, and the vibrator applies vibration in an angle range of-15 degrees to 15 degrees with the exhaust surface of the gas explosion plate when vibration radiation is carried out, and preferably the angle formed by the vibrator and the exhaust surface of the gas explosion plate is 0 degree; and/or

The explosion plate is prepared from a nano activated carbon fiber material, micropores are arranged on the nano activated carbon fiber, and the pore diameter of each micropore is less than 1.5 mu m, preferably 0.01 to 1.5 mu m, and more preferably 0.1 to 1.0 mu m.