Deuterium-depleted water composition as well as preparation method and application thereof

Abstract

The invention provides a deuterium-depleted water composition as well as a preparation method and application thereof, and belongs to the technical field of medicines. Comprising the following steps: S1, preparing deuterium-depleted water; s2, extracting active components of the traditional Chinese medicines; s3, activating the strain; s4, assisted fermentation with deuterium-depleted water; and S5, preparing the deuterium-depleted water composition. The deuterium-depleted water composition prepared by the invention has better effects of increasing the activity value of NK (Natural Killer) cells and improving body immune disorder, so that the human immune function is enhanced, sediments are removed, molecular dissolution is promoted, blood vessels are dredged, and the blood circulation function of each system is improved, and therefore, many diseases such as hypertension, cerebral thrombosis and the like are not easy to generate or worsen; the generation of diseases or the morbidity after the generation of the diseases is reduced, the antioxidant and anti-inflammatory capacities are improved, and better anti-tumor activity is realized.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a deuterium-depleted water composition and a preparation method and application thereof. Background Art

[0002] Deuterium depleted water, or DDW for short, is water with a deuterium content lower than the natural level. Deuterium is a naturally occurring heavy isotope of hydrogen, mainly present in natural water in the form of semi-deuterium water (HDO). In the human body, one of the signals for cell division is a temporary increase in the deuterium level relative to the hydrogen level, i.e. an increase in the deuterium / hydrogen ratio (D / H), so deuterium is essential for cell reproduction.

[0003] The deuterium concentration in the human body ranges from 110-120ppm. This value seems insignificant, but its amount in the body is 6 times that of calcium (Ca), 10 times that of magnesium (Mg), and 3 times that of potassium (K). The amount of deuterium is crucial to the human body. Long-term drinking of heavy water with high deuterium content has adverse effects and harmful effects on the human body's metabolism and enzyme system. Additional stress is generated in the DNA helix structure, causing chaos in the DNA helix arrangement, and even resynthesis, resulting in gene mutations. my country opposes Japan's discharge of nuclear wastewater into the sea because the content of harmful elements such as deuterium in its wastewater is too high.

[0004] According to statistics from the World Health Organization, by 2030, the annual number of cancer deaths is expected to exceed 13 million. It can be seen that even if the cost of anti-cancer drugs and treatments has increased exponentially, the efficacy is still minimal. Faced with such a huge number of lives, the overall focus and direction of cancer treatment needs to be changed. Until the new cellular submolecular method based on the change in the ratio of deuterium to hydrogen, "deuterium-depleted water (DDW) therapy", was discovered, the place shrouded in clouds seemed to have ushered in a long-lost light. Moreover, from the perspective of cancer metabolism theory, drinking deuterium-depleted water can help reduce the deuterium concentration in each body cell and prevent the deuterium concentration from exceeding the normal threshold level, thereby compensating for the damage or loss of mitochondrial function in tumor cells, thereby reducing the frequency of uncontrolled cell division; from the perspective of cancer genetics theory, deuterium-depleted water can offset the process of stimulating cell division at the molecular level triggered by genetic differences.

[0005] The advantage of deuterium-depleted water over conventional treatment is that it can significantly reduce the occurrence of adverse reactions and improve tolerance, and can maximize the maintenance of the body's own immune system. Nevertheless, the effect of deuterium-depleted water is more on intervention when the disease relapses, and on the alleviation of the disease, delay of deterioration, and extension of life of cancer patients, rather than fundamentally curing such diseases. It can only be used as an effective auxiliary means of cancer prevention and treatment. In addition, research results have shown that changing the deuterium level can reduce the susceptibility to depression, improve long-term memory, and have radiation protection and anti-aging effects.

[0006] The traditional sources and production methods of deuterium-depleted water are:

[0007] 1. Glacier water is natural low-deuterium water, with a deuterium content of 135ppm;

[0008] 2. Deuterium-depleted water can be produced by water distillation. Since water and heavy water have different boiling points, deuterium-depleted water can be produced by water distillation.

[0009] Glaciers are located on a plateau far from human habitation, so the cost of mining and transportation is huge. In addition, the deuterium content is not much different from that of ordinary water, so there has been no large-scale mining and utilization so far. The water distillation method also consumes a lot of energy because the boiling point difference between water and heavy water is very small, so there is no large-scale production. It only produces low-deuterium water as a by-product in the production of oxygen-18, and the output is very limited.

[0010] Therefore, providing a method with low energy consumption, low production cost, and the ability to achieve industrial mass production of deuterium-depleted water will have broad application prospects. Summary of the invention

[0011] The purpose of the present invention is to propose a deuterium-depleted water composition and its preparation method and application, which has the advantages of improving the activity value of NK cells, improving the body's immune disorders, thereby enhancing the body's immune function, removing sediments, promoting molecular dissolution, opening up blood vessels, and improving the blood circulation function of various systems, thereby making it difficult for many diseases such as hypertension and cerebral thrombosis to occur or deteriorate, reducing the occurrence of diseases or the incidence of diseases after occurrence, improving antioxidant and anti-inflammatory capabilities, and having good anti-tumor activity.

[0012] The technical solution of the present invention is achieved in this way:

[0013] The present invention provides a method for preparing a deuterium-depleted water composition, comprising the following steps:

[0014] S1. Preparation of deuterium-depleted water: a hydrophobic catalyst is filled into an exchange column, preheated mineral water is introduced into the column from the top of the exchange column, and preheated high-purity hydrogen is introduced from the bottom of the exchange column. In the exchange column, a catalytic exchange reaction occurs between the high-purity hydrogen and water, and the product deuterium-depleted water is output from the bottom. The deuterium-depleted water is collected. After the production is completed, the exchange column is flushed and cleaned with high-purity nitrogen to regenerate the filler;

[0015] S2. Extraction of active components of traditional Chinese medicine: Mix coptis root, poria, ginseng and Panax notoginseng, crush them, add low-deuterium water and heat to boiling for extraction, repeat 2-3 times, filter, retain the solid, combine the filtrate, concentrate, and dry to obtain the active components of traditional Chinese medicine;

[0016] S3. Activation of bacteria: Selenium-enriched yeast and Lactobacillus plantarum were inoculated into Gao's medium, activated and cultured to obtain a culture medium with a bacterial content of 10 8 -10 9 cfu / mL of bacterial seed solution;

[0017] S4. Deuterium-depleted water-assisted fermentation: adding the solid in step S2 to deuterium-depleted water, sterilizing, inoculating activated selenium-enriched yeast and Lactobacillus plantarum seed liquid, fermenting, filtering, concentrating the filtrate, and freeze-drying to obtain a fermentation component;

[0018] S5. Preparation of deuterium-depleted water composition: Evenly mix the active components of traditional Chinese medicine, fermentation components and deuterium-depleted water to prepare the deuterium-depleted water composition.

[0019] As a further improvement of the present invention, the temperature of the preheated deionized water in step S1 is 40-60°C, the temperature of the preheated high-purity hydrogen is 40-60°C, the purity is >99.99%, the gas-liquid molar ratio of the deionized water and hydrogen is 0.4-0.7:1, the deuterium content of the low-deuterium water is ≤125ppm, and the half-peak width is <70Hz.

[0020] As a further improvement of the present invention, the preparation method of the hydrophobic catalyst is as follows:

[0021] T1. Preparation of Pt / Pd@graphene: dissolve palladium chloride in hydrochloric acid to obtain H2PdCl4 solution, add chloroplatinic acid, stir and mix evenly, add graphene oxide, stir and react, dry, wash, pass hydrogen to heat and reduce, then pass hydrazine hydrate vapor to reduce, and obtain Pt / Pd@graphene;

[0022] T2. Preparation of hydrophobic catalyst: dissolving fluorine-containing silane in an organic solvent to obtain an oil phase; adding a porogen, an emulsifier and Pt / Pd@graphene into water to obtain a suspension; dropping the suspension into the oil phase, emulsifying, adjusting the pH value of the solution, stirring the reaction, centrifuging, washing, spray drying, and obtaining a hydrophobic catalyst.

[0023] As a further improvement of the present invention, the concentration of the hydrochloric acid in step T1 is 0.3-0.5 mol / L, the mass ratio of the palladium chloride, chloroplatinic acid, and graphene oxide is 3-5:7-12:30-50, the stirring reaction time is 12-15 h, the temperature of the hydrogen heating reduction reaction is 300-350 ° C, the time is 2-4 h, and the time of the hydrazine hydrate steam reduction is 3-5 h.

[0024] As a further improvement of the present invention, the fluorine-containing silane in step T2 is selected from 1H, 1H, 2H, 2H-perfluorodecyltriethoxysilane, 1H, 1H, 2H, 2H-perfluorodecyltrimethoxysilane, dodecafluoroheptylpropyltrimethoxysilane, dodecafluoroheptylpropylmethyldimethoxysilane, 3,3,3-trifluoropropylmethyldimethoxysilane, 3,3,3-trifluoropropyltrimethoxysilane, 1H, 1H, 2H, 2H-perfluorooctyltriethoxysilane or 1H, 1H, 2H, 2H-perfluorooctyltrimethoxysilane, the mass ratio of the fluorinated silane, the porogen, the emulsifier and Pt / Pd@graphene is 12-15:0.5-1:0.2-0.4:6-8, the porogen is hexadecyltrimethylammonium chloride, the emulsifier is selected from at least one of Tween-20, Tween-40, Tween-60, Tween-80 and Tween-85, the pH value of the solution is adjusted to 9-10, and the stirring reaction time is 5-7h.

[0025] As a further improvement of the present invention, in step S2, the mass ratio of coptis root, poria, ginseng, notoginseng and deuterium-depleted water is 3-7:2-4:1-2:3-5:100, and the time for heating to boiling extraction is 3-5h.

[0026] As a further improvement of the present invention, the conditions for the activation culture of the selenium-enriched yeast in step S3 are 45-50°C, 100-200r / min, and activation culture for 18-24h, and the conditions for the activation culture of the plant lactobacillus are 35-40°C, 100-200r / min, and activation culture for 18-24h under anaerobic conditions.

[0027] As a further improvement of the present invention, the mass ratio of the solid and the deuterium-depleted water in step S4 is 10-15:100, the inoculation amounts of the activated selenium-enriched yeast and plant lactobacillus species seed liquid are 2-4v / v% and 1-2v / v%, respectively, the fermentation and culture conditions are under micro-anoxic conditions, 38-42°C, 100-200r / min, and the fermentation and culture is 36-48h, the micro-anoxic conditions are oxygen content 5-10v / v%, carbon dioxide content 3-7v / v%, and the balance is nitrogen; the mass ratio of the active components of the traditional Chinese medicine, the fermentation components and the deuterium-depleted water in step S5 is 2-3:1-2:10.

[0028] The present invention further protects a deuterium-deficient water composition prepared by the above preparation method.

[0029] The present invention further protects the use of the above-mentioned deuterium-deficient water composition in the preparation of anti-tumor, anti-inflammatory and antioxidant drugs.

[0030] The present invention has the following beneficial effects:

[0031] The invention prepares a hydrophobic catalyst. Pt and Pd are loaded on the surface of graphene oxide by an impregnation method, and Pt / Pd@graphene is obtained by reduction. Then, a water suspension prepared by dissolving fluorine-containing silane in an oil phase, and a porogen, an emulsifier and Pt / Pd@graphene is dripped into the oil phase for emulsification to form an oil-in-water emulsion. The fluorine-containing group extends into the oil phase of an outer layer, and the inner layer wraps the Pt / Pd@graphene, thereby obtaining a hydrophobic catalyst containing fluorine on the surface. The Pt and Pd bimetallic catalyst is used, the catalytic efficiency is greatly improved, and a synergistic effect is achieved. Compared with a common carrier, the graphene oxide carrier has a large specific surface area, and the loading amount of the catalytic metal is greatly improved, thereby improving the catalytic activity. Due to its hydrophobic structure, the catalyst can block the contact between liquid water and active metals in low-temperature environments, thereby preventing gaseous reactants such as hydrogen (whose solubility and diffusion rate are relatively low in liquid) and vapor-phase water from contacting the active sites of the catalyst, thus avoiding the phenomenon of catalyst "poisoning".

[0032] The catalytic exchange process mainly includes two parts: gas-liquid phase conversion and gas-phase catalytic exchange. Taking H2-HDO isotope exchange as an example, the exchange process is as follows:

[0033]

[0034] The process can be divided into two steps:

[0035]

[0036] Among them, g means gas phase, l represents liquid phase, and v represents vapor phase.

[0037] Equation 2 represents the transition process between gas and liquid, while Equation 3 represents the gas phase catalytic conversion reaction.

[0038] The method of the present invention is used to simply and conveniently prepare low-deuterium water, and is easy to realize industrial application, and the catalyst regeneration is simple and convenient for repeated use. After continuous detection, the deuterium content is low, the deuterium content is ≤122PPM (ordinary water>150ppm), the half-peak width is <70Hz (ordinary water 100-150Hz), and the water contains strontium, selenium, lithium, potassium, metasilicate rare mineral elements and weak alkalinity that are beneficial to human health. The water does not contain heavy metals and other harmful bacteria that are harmful to the human body. The mineral water comes from a zero-pollution water source in the original ecological environment, has a moderate mineralization, and does not cause a burden on kidney metabolism when drunk for a long time.

[0039] The deuterium-depleted water prepared by the present invention has no free charge and is a very stable water with a small molecular cluster structure, and has strong penetration, dissolving power, emulsifying power, metabolic power and activation power. It can activate the function of human cells, enhance human immunity, improve body tolerance, promote blood lipid dissolution, and provide a good basic condition for defeating cancer. In addition, the growth mechanism of cancer cells is closely related to deuterium atoms. By reducing the concentration of deuterium in the body, the growth of cancer cells in the body can be inhibited and the apoptosis of cancer cells can be induced.

[0040] Coptis chinensis is rich in berberine, Poria cocos is rich in tuckahoe polysaccharide, tuckahoe saponin, etc., ginsenosides in ginseng, ginseng polysaccharide and notoginseng total saponin in Panax notoginseng have excellent anti-inflammatory and antioxidant effects, and have a good inhibitory effect on tumor cells, thus playing a good anti-tumor role. Because the surface tension of deuterium-depleted water is different from that of distilled water, deuterium-depleted water has better penetration and solubility than ordinary water, which helps to dissolve and extract active components in plant cells, thereby greatly improving the extraction rate and efficacy of active components of traditional Chinese medicine.

[0041] In addition, deuterium-depleted water can increase the activity value of probiotics, significantly promote enzyme reactions, improve basal metabolism, help activate the body's probiotic cells, increase cell age, prolong survival time, maintain cell growth rate, and improve cell DNA synthesis ability, promote cell proliferation, thereby greatly improving the vitality of probiotics and improving its fermentation and extraction ability of the substrate Chinese medicine solid residue, thereby obtaining more active components and improving the anti-tumor, antioxidant and anti-inflammatory effects of the components.

[0042] Cancer cells need deuterium at the beginning of the division cycle. If there is not enough deuterium, the metabolism of cancer cells will be disordered and they will not be able to continue to divide. When the deuterium content is lower than 110ppm, cancer cells will die. Low-deuterium water can inhibit the heavy hydrogen environment required for cancer cell growth, increase blood oxygen content and oxygen absorption, activate the vitality of human tissue cells, and improve cell immunity. Therefore, it can achieve the purpose of reducing tumor index values, inhibiting the proliferation of deteriorated cells and cancer cells, and preventing cancer and health care.

[0043] The deuterium-depleted water composition prepared by the present invention has the advantages of improving the activity value of NK cells, improving the body's immune disorders, thereby enhancing the body's immune function, removing deposits, promoting molecular dissolution, unblocking blood vessels, and improving the blood circulation function of various systems, thereby making it difficult for many diseases such as hypertension and cerebral thrombosis to occur or deteriorate, reducing the occurrence of diseases or the incidence of diseases after occurrence, improving antioxidant and anti-inflammatory capabilities, and having good anti-tumor activity. DETAILED DESCRIPTION

[0044] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0045] Graphene oxide, with a thickness of <5 nm, an oxygen content of 30-35%, and a sheet diameter of 18-20 μm, was purchased from Jiangsu Xianfeng Nanomaterial Technology Co., Ltd.

[0046] The mineral water is Wulongquan mineral water from Changbai Mountain in Baishan City, Jilin Province.

[0047] Selenium-enriched yeast, 10 billion cfu / g, was purchased from Angel Yeast Co., Ltd.; Lactobacillus plantarum, 20 billion cfu / g, was purchased from Shandong Junle Biotechnology Co., Ltd.

[0048] Preparation Example 1 Preparation of hydrophobic catalyst

[0049] Here’s how:

[0050] T1. Preparation of Pt / Pd@graphene: 0.3 g palladium chloride was dissolved in 100 mL 0.3 mol / L hydrochloric acid to prepare H2PdCl4 solution, 0.7 g chloroplatinic acid was added, stirred and mixed for 10 min, 3 g graphene oxide was added, stirred and reacted for 12 h, dried, washed, heated to 300 ° C for reduction for 2 h, and then reduced by hydrazine hydrate vapor at 150 ° C for 3 h to obtain Pt / Pd@graphene;

[0051] T2. Preparation of a hydrophobic catalyst: 12 g of 3,3,3-trifluoropropylmethyldimethoxysilane was dissolved in 500 mL of ethyl acetate to obtain an oil phase; 0.5 g of hexadecyltrimethylammonium chloride, 0.2 g of Tween-20 and 6 g of Pt / Pd@graphene were added to water to obtain a suspension; the suspension was added dropwise to the oil phase, emulsified at 7000 r / min for 15 min, the pH value of the solution was adjusted to 9, the reaction was stirred for 5 h, centrifuged, washed, and spray-dried to obtain a hydrophobic catalyst.

[0052] Preparation Example 2 Preparation of hydrophobic catalyst

[0053] Here’s how:

[0054] T1. Preparation of Pt / Pd@graphene: 0.5 g palladium chloride was dissolved in 100 mL 0.5 mol / L hydrochloric acid to prepare H2PdCl4 solution, 1.2 g chloroplatinic acid was added, stirred and mixed for 10 min, 5 g graphene oxide was added, stirred and reacted for 15 h, dried, washed, heated to 350 ° C for 4 h, and then reduced by hydrazine hydrate vapor at 150 ° C for 4 h to obtain Pt / Pd@graphene;

[0055] T2. Preparation of hydrophobic catalyst: 15 g of dodecafluoroheptylpropylmethyldimethoxysilane was dissolved in 500 mL of ethyl acetate to obtain an oil phase; 1 g of hexadecyltrimethylammonium chloride, 0.4 g of Tween-40 and 8 g of Pt / Pd@graphene were added to water to obtain a suspension; the suspension was added dropwise to the oil phase, emulsified at 7000 r / min for 15 min, the pH value of the solution was adjusted to 10, the reaction was stirred for 7 h, centrifuged, washed, and spray-dried to obtain a hydrophobic catalyst.

[0056] Preparation Example 3 Preparation of hydrophobic catalyst

[0057] Here’s how:

[0058] T1. Preparation of Pt / Pd@graphene: 0.4 g palladium chloride was dissolved in 100 mL 0.4 mol / L hydrochloric acid to prepare H2PdCl4 solution, 1 g chloroplatinic acid was added, stirred and mixed for 10 min, 4 g graphene oxide was added, stirred and reacted for 13 h, dried, washed, heated to 320 ° C for 3 h, and then reduced by hydrazine hydrate vapor at 150 ° C for 5 h to obtain Pt / Pd@graphene;

[0059] T2. Preparation of hydrophobic catalyst: 13 g of dodecafluoroheptylpropyltrimethoxysilane was dissolved in 500 mL of ethyl acetate to obtain an oil phase; 0.7 g of hexadecyltrimethylammonium chloride, 0.3 g of Tween-85 and 7 g of Pt / Pd@graphene were added to water to obtain a suspension; the suspension was added dropwise to the oil phase, emulsified at 7000 r / min for 15 min, the pH value of the solution was adjusted to 9.5, the reaction was stirred for 6 h, centrifuged, washed, and spray-dried to obtain a hydrophobic catalyst.

[0060] Comparative Preparation Example 1

[0061] Compared with Preparation Example 3, the difference is that H2PdCl4 is not added in Step T1.

[0062] The details are as follows:

[0063] T1. Preparation of Pt@graphene: 1.4 g of chloroplatinic acid was dissolved in 100 mL of water, 4 g of graphene oxide was added, stirred and reacted for 13 h, dried, washed, introduced with hydrogen and heated to 320 °C for reduction for 3 h, and then reduced with hydrazine hydrate vapor at 150 °C for 5 h to obtain Pt@graphene;

[0064] T2. Preparation of hydrophobic catalyst: 13 g of dodecafluoroheptylpropyltrimethoxysilane was dissolved in 500 mL of ethyl acetate to obtain an oil phase; 0.7 g of hexadecyltrimethylammonium chloride, 0.3 g of Tween-85 and 7 g of Pt@graphene were added to water to obtain a suspension; the suspension was added dropwise to the oil phase, emulsified at 7000 r / min for 15 min, the pH value of the solution was adjusted to 9.5, the reaction was stirred for 6 h, centrifuged, washed, and spray-dried to obtain a hydrophobic catalyst.

[0065] Comparative Preparation Example 2

[0066] Compared with Preparation Example 3, the difference is that chloroplatinic acid is not added in step T1.

[0067] The details are as follows:

[0068] T1. Preparation of Pd@graphene: 1.4 g palladium chloride was dissolved in 100 mL 0.4 mol / L hydrochloric acid to obtain H2PdCl4 solution, stirred and mixed for 10 min, 4 g graphene oxide was added, stirred and reacted for 13 h, dried, washed, heated to 320 ° C for 3 h, and then reduced by hydrazine hydrate vapor at 150 ° C for 5 h to obtain Pd@graphene;

[0069] T2. Preparation of hydrophobic catalyst: 13 g of dodecafluoroheptylpropyltrimethoxysilane was dissolved in 500 mL of ethyl acetate to obtain an oil phase; 0.7 g of hexadecyltrimethylammonium chloride, 0.3 g of Tween-85 and 7 g of Pd@graphene were added to water to obtain a suspension; the suspension was added dropwise to the oil phase, emulsified at 7000 r / min for 15 min, the pH value of the solution was adjusted to 9.5, the reaction was stirred for 6 h, centrifuged, washed, and spray-dried to obtain a hydrophobic catalyst.

[0070] Comparative Preparation Example 3

[0071] Compared with Preparation Example 3, the difference is that the fluorine-containing silane in step T2 is replaced by tetraethyl orthosilicate.

[0072] The details are as follows:

[0073] T1. Preparation of Pt / Pd@graphene: 0.4 g palladium chloride was dissolved in 100 mL 0.4 mol / L hydrochloric acid to prepare H2PdCl4 solution, 1 g chloroplatinic acid was added, stirred and mixed for 10 min, 4 g graphene oxide was added, stirred and reacted for 13 h, dried, washed, heated to 320 ° C for 3 h, and then reduced by hydrazine hydrate vapor at 150 ° C for 5 h to obtain Pt / Pd@graphene;

[0074] T2. Preparation of catalyst: 13 g of tetraethyl orthosilicate was dissolved in 500 mL of ethyl acetate to obtain an oil phase; 0.7 g of hexadecyltrimethylammonium chloride, 0.3 g of Tween-85 and 7 g of Pt / Pd@graphene were added into water to obtain a suspension; the suspension was added dropwise into the oil phase, emulsified at 7000 r / min for 15 min, the pH value of the solution was adjusted to 9.5, the reaction was stirred for 6 h, centrifuged, washed, and spray-dried to obtain the catalyst.

[0075] Comparative Preparation Example 4

[0076] Compared with Preparation Example 3, the difference is that the porogen hexadecyltrimethylammonium chloride is not added in step T2.

[0077] The details are as follows:

[0078] T1. Preparation of Pt / Pd@graphene: 0.4 g palladium chloride was dissolved in 100 mL 0.4 mol / L hydrochloric acid to obtain H2PdCl4 solution, 1 g chloroplatinic acid was added, stirred and mixed for 10 min, 4 g graphene oxide was added, stirred and reacted for 13 h, dried, washed, and a mixed gas of hydrogen and hydrazine hydrate vapor was introduced, wherein the volume ratio of hydrogen and hydrazine hydrate vapor was 8:3, heated to 320°C, and reduced for 3 h to obtain Pt / Pd@graphene;

[0079] T2. Preparation of hydrophobic catalyst: 13 g of dodecafluoroheptylpropyltrimethoxysilane was dissolved in 500 mL of ethyl acetate to obtain an oil phase; 1 g of Tween-85 and 7 g of Pt / Pd@graphene were added to water to obtain a suspension; the suspension was added dropwise to the oil phase, emulsified at 7000 r / min for 15 min, the pH value of the solution was adjusted to 9.5, the reaction was stirred for 6 h, centrifuged, washed, and spray-dried to obtain a hydrophobic catalyst.

[0080] Test Example 1

[0081] N2 low temperature physical adsorption-desorption test: The specific surface area (m2) of the catalysts prepared in Preparation Examples 1-3 or Comparative Preparation Examples 1-4 was tested using a fully automatic physical / chemical adsorption analyzer. 2 / g), and tested its water contact angle (°). The results are shown in Table 1.

[0082] Table 1

[0083] Group <![CDATA[Specific surface area (m 2 / g)]]> Water contact angle (°) Preparation Example 1 325 122 Preparation Example 2 322 124 Preparation Example 3 329 125 Comparative Preparation Example 1 310 120 Comparative Preparation Example 2 304 118 Comparative Preparation Example 3 320 83 Comparative Preparation Example 4 282 109

[0084] It can be seen from the above table that the hydrophobic catalysts prepared in Preparation Examples 1-3 of the present invention have a relatively high specific surface area.

[0085] Example 1

[0086] This embodiment provides a method for preparing a deuterium-depleted water composition, comprising the following steps:

[0087] S1. Preparation of deuterium-depleted water: The hydrophobic catalyst prepared in Preparation Example 1 is filled into an exchange column, mineral water preheated to 40° C. is introduced into the column from the top of the exchange column, and high-purity hydrogen (purity> 99.99%) preheated to 40° C. is introduced from the bottom of the exchange column. In the exchange column, a catalytic exchange reaction occurs between high-purity hydrogen and water, and the gas-liquid molar ratio of the deionized water and hydrogen is 0.4:1. The product deuterium-depleted water is output from the bottom, and the deuterium-depleted water is collected. The deuterium content of the deuterium-depleted water is 110 ppm, and the half-peak width is 65 Hz. After the production is completed, the exchange column is rinsed and cleaned with high-purity nitrogen to regenerate the filler;

[0088] S2. Extraction of active components of traditional Chinese medicine: 3g of Coptis chinensis, 2g of Poria cocos, 1g of ginseng, and 3g of Panax notoginseng were mixed, crushed, and 100g of low-deuterium water was added and heated to boiling for 3h, and the mixture was repeated twice, filtered, the solid was retained, the filtrate was combined, concentrated, and dried to obtain active components of traditional Chinese medicine;

[0089] S3. Activation of bacteria: Selenium-enriched yeast and Lactobacillus plantarum were inoculated into Gao's medium, activated and cultured to obtain a culture medium with a bacterial content of 10 8 -10 9 cfu / mL of bacterial seed solution;

[0090] The conditions for the activation culture of the selenium-enriched yeast are 45° C., 100 r / min, and activation culture for 20 hours;

[0091] The activation culture conditions of the plant lactobacillus pair are anaerobic conditions, 35° C., 100 r / min, and activation culture for 20 hours;

[0092] S4. Fermentation with deuterium-depleted water: 10 g of the solid in step S2 was added to 100 g of deuterium-depleted water, sterilized, inoculated with activated selenium-enriched yeast and Lactobacillus plantarum seed liquid, the inoculation amounts were 2 v / v% and 1 v / v%, respectively, under slightly hypoxic conditions, 38 ° C, 100 r / min, fermentation culture for 36 h, filtered, the filtrate was concentrated, freeze-dried, and fermentation components were obtained;

[0093] The micro-anoxia condition is an oxygen content of 5 v / v%, a carbon dioxide content of 3 v / v%, and the balance is nitrogen;

[0094] S5. Preparation of deuterium-depleted water composition: 2 g of active components of traditional Chinese medicine, 1 g of fermentation components and 10 g of deuterium-depleted water were mixed for 10 min, filtered, and a deuterium-depleted water composition was obtained.

[0095] Example 2

[0096] This embodiment provides a method for preparing a deuterium-depleted water composition, comprising the following steps:

[0097] S1. Preparation of deuterium-depleted water: The hydrophobic catalyst prepared in Preparation Example 2 is filled into an exchange column, mineral water preheated to 60° C. is introduced into the column from the top of the exchange column, and high-purity hydrogen (purity> 99.99%) preheated to 60° C. is introduced from the bottom of the exchange column. In the exchange column, a catalytic exchange reaction occurs between high-purity hydrogen and water, and the gas-liquid molar ratio of the deionized water and hydrogen is 0.7:1. The product deuterium-depleted water is output from the bottom, and the deuterium-depleted water is collected. The deuterium content of the deuterium-depleted water is 109 ppm, and the half-peak width is 62 Hz. After the production is completed, the exchange column is rinsed and cleaned with high-purity nitrogen to regenerate the filler;

[0098] S2. Extraction of active components of traditional Chinese medicine: 7g of Coptis chinensis, 4g of Poria cocos, 2g of ginseng, and 5g of Panax notoginseng were mixed, crushed, and 100g of low-deuterium water was added and heated to boiling for 5h, and the mixture was repeated 3 times, filtered, the solid was retained, the filtrate was combined, concentrated, and dried to obtain active components of traditional Chinese medicine;

[0099] S3. Activation of bacteria: Selenium-enriched yeast and Lactobacillus plantarum were inoculated into Gao's medium, activated and cultured to obtain a culture medium with a bacterial content of 10 8 -10 9 cfu / mL of bacterial seed solution;

[0100] The conditions for the activation culture of the selenium-enriched yeast are 50° C., 200 r / min, and the activation culture is 20 h;

[0101] The activation culture conditions of the plant lactobacillus pair are anaerobic conditions, 40° C., 200 r / min, and activation culture for 20 hours;

[0102] S4. Fermentation with deuterium-depleted water: 15 g of the solid in step S2 was added to 100 g of deuterium-depleted water, sterilized, and inoculated with activated selenium-enriched yeast and Lactobacillus plantarum seed liquid, with inoculation amounts of 4 v / v% and 2 v / v%, respectively, under slightly hypoxic conditions, at 42 ° C, 200 r / min, fermentation culture for 48 h, filtered, the filtrate was concentrated, and freeze-dried to obtain a fermentation component;

[0103] The micro-anoxic condition is an oxygen content of 10 v / v%, a carbon dioxide content of 7 v / v%, and the balance is nitrogen;

[0104] S5. Preparation of deuterium-depleted water composition: 3 g of active components of traditional Chinese medicine, 2 g of fermentation components and 10 g of deuterium-depleted water were mixed for 10 min, filtered, and a deuterium-depleted water composition was prepared.

[0105] Example 3

[0106] This embodiment provides a method for preparing a deuterium-depleted water composition, comprising the following steps:

[0107] S1. Preparation of deuterium-depleted water: The hydrophobic catalyst prepared in Preparation Example 3 is filled into an exchange column, mineral water preheated to 50° C. is introduced into the column from the top of the exchange column, and high-purity hydrogen (purity> 99.99%) preheated to 50° C. is introduced from the bottom of the exchange column. In the exchange column, a catalytic exchange reaction occurs between high-purity hydrogen and water, and the gas-liquid molar ratio of the deionized water and hydrogen is 0.55:1. The product deuterium-depleted water is output from the bottom, and the deuterium-depleted water is collected. The deuterium content of the deuterium-depleted water is 102 ppm, and the half-peak width is 60 Hz. After the production is completed, the exchange column is rinsed and cleaned with high-purity nitrogen to regenerate the filler;

[0108] S2. Extraction of active components of traditional Chinese medicine: 5g of Coptis chinensis, 3g of Poria cocos, 1.5g of ginseng, and 4g of Panax notoginseng were mixed, crushed, and 100g of low-deuterium water was added and heated to boiling for 4h, and the mixture was repeated 3 times, filtered, the solid was retained, the filtrate was combined, concentrated, and dried to obtain active components of traditional Chinese medicine;

[0109] S3. Activation of bacteria: Selenium-enriched yeast and Lactobacillus plantarum were inoculated into Gao's medium, activated and cultured to obtain a culture medium with a bacterial content of 10 8 -10 9 cfu / mL of bacterial seed solution;

[0110] The conditions for the activation culture of the selenium-enriched yeast are 47° C., 150 r / min, and activation culture for 20 hours;

[0111] The activation culture conditions of the plant lactobacillus pair are anaerobic conditions, 37° C., 150 r / min, and activation culture for 20 hours;

[0112] S4. Deuterium-depleted water-assisted fermentation: 12 g of the solid in step S2 was added to 100 g of deuterium-depleted water, sterilized, inoculated with activated selenium-enriched yeast and Lactobacillus plantarum seed liquid, the inoculation amounts were 3 v / v% and 1.5 v / v%, respectively, under slightly hypoxic conditions, 40 ° C, 150 r / min, fermentation culture for 42 h, filtered, the filtrate was concentrated, freeze-dried, and the fermentation component was obtained;

[0113] The micro-anoxia condition is an oxygen content of 7 v / v%, a carbon dioxide content of 5 v / v%, and the balance is nitrogen;

[0114] S5. Preparation of deuterium-depleted water composition: 2.5 g of active components of traditional Chinese medicine, 1.2 g of fermentation components and 10 g of deuterium-depleted water were mixed for 10 min, filtered, and a deuterium-depleted water composition was obtained.

[0115] Comparative Examples 1-4

[0116] Compared with Example 3, the difference is that the hydrophobic catalysts are prepared from Comparative Preparation Examples 1-4 respectively.

[0117] Comparative Example 5

[0118] Compared with Example 3, the difference is that the deuterium-depleted water in step S2 is replaced by ordinary distilled water.

[0119] The details are as follows:

[0120] S2. Extraction of active components of traditional Chinese medicine: Mix 5g of Coptis chinensis, 3g of Poria cocos, 1.5g of ginseng, and 4g of Panax notoginseng, grind them, add 100g of ordinary distilled water and heat to boiling for 4h, repeat 3 times, filter, retain the solid, combine the filtrate, concentrate, and dry to obtain the active components of traditional Chinese medicine.

[0121] Comparative Example 6

[0122] Compared with Example 3, the difference is that the deuterium-depleted water in step S4 is replaced by ordinary distilled water.

[0123] The details are as follows:

[0124] S4. Deuterium-depleted water-assisted fermentation: 12 g of the solid in step S2 was added to 100 g of ordinary distilled water, sterilized, inoculated with activated selenium-enriched yeast and Lactobacillus plantarum seed liquid, the inoculation amounts were 3 v / v% and 1.5 v / v%, respectively, under slightly hypoxic conditions, 40 ° C, 150 r / min, fermentation culture for 42 h, filtered, the filtrate was concentrated, freeze-dried, and the fermentation component was obtained;

[0125] The micro-anoxic condition is an oxygen content of 7 v / v%, a carbon dioxide content of 5 v / v%, and the balance is nitrogen.

[0126] Comparative Example 7

[0127] Compared with Example 3, the difference is that no selenium-enriched yeast seed liquid is inoculated in step S4.

[0128] The details are as follows:

[0129] S4. Fermentation with deuterium-depleted water: 12 g of the solid in step S2 was added to 100 g of deuterium-depleted water, sterilized, inoculated with activated Lactobacillus plantarum seed solution, the inoculation amount was 4.5 v / v%, under slightly anoxic conditions, 40 ° C, 150 r / min, fermented for 42 h, filtered, the filtrate was concentrated, and freeze-dried to obtain a fermentation component;

[0130] The micro-anoxic condition is an oxygen content of 7 v / v%, a carbon dioxide content of 5 v / v%, and the balance is nitrogen.

[0131] Comparative Example 8

[0132] Compared with Example 3, the difference is that the Lactobacillus plantarum seed liquid is not inoculated in step S4.

[0133] The details are as follows:

[0134] S4. Deuterium-depleted water-assisted fermentation: 12 g of the solid in step S2 was added to 100 g of deuterium-depleted water, sterilized, and inoculated with activated selenium-enriched yeast seed liquid at an inoculation rate of 4.5 v / v%, under slightly anoxic conditions, at 40° C., 150 r / min, and fermented for 42 h, filtered, the filtrate was concentrated, and freeze-dried to obtain a fermentation component;

[0135] The micro-anoxic condition is an oxygen content of 7 v / v%, a carbon dioxide content of 5 v / v%, and the balance is nitrogen.

[0136] Comparative Example 9

[0137] Compared with Example 3, the difference is that no active components of traditional Chinese medicine are added in step S5.

[0138] The details are as follows:

[0139] S5. Preparation of deuterium-depleted water composition: 3.7 g of fermentation components and 10 g of deuterium-depleted water were mixed for 10 min, and filtered to obtain a deuterium-depleted water composition.

[0140] Comparative Example 10

[0141] Compared with Example 3, the difference is that no fermentation component is added in step S5.

[0142] The details are as follows:

[0143] S5. Preparation of deuterium-depleted water composition: 3.7 g of active ingredients of traditional Chinese medicine and 10 g of deuterium-depleted water were mixed for 10 min, filtered, and a deuterium-depleted water composition was prepared.

[0144] Test Example 1 Effect on Cell Cycle

[0145] Take the deuterium-depleted water compositions prepared in Examples 1-3 and Comparative Examples 1-10 respectively, and the control group is ordinary distilled water. Take HepG2 (human liver cancer cells) and culture them in DMEM high-glucose culture medium containing 10% fetal bovine serum and 1% double antibody, and grow. The environment is 37°C, 5% CO2 in an incubator, digest the cells with trypsin, collect the cells after trypsin digestion, centrifuge, discard the supernatant, add pre-cooled PBS, resuspend the cells, and centrifuge. Add PBS to fully resuspend, add pre-cooled anhydrous ethanol dropwise to a final concentration of 75%, and fix at 4°C for 24h. Centrifuge the fixed cell suspension, discard the supernatant, add 1mL pre-cooled PBS, resuspend the cells, centrifuge again, and precipitate the cells. The configuration of the staining working solution is mixed according to the ratio of staining buffer: propidium iodide PI staining solution (20X): RNaseA (50X) = 100:5:2, add 500μL of staining working solution to each tube of sample, and warm it at 37°C in the dark for 30min. After staining, the samples were stored at 4°C away from light, tested, and analyzed. The results are shown in Table 2.

[0146] Table 2

[0147]

[0148]

[0149] The whole process that cells go through from the completion of one division to the end of the next division is called the cell cycle. The period from mitosis to DNA replication is called the presynthesis phase (G1 phase), which is to prepare materials and energy for DNA replication in the next stage S phase. The S phase is the DNA synthesis phase, and the enzymes required for DNA replication are synthesized during this period. The G2 phase is the DNA synthesis late phase, during which DNA synthesis stops, and RNA and protein are synthesized in large quantities. As shown in the above table, compared with the control group, the low-deuterium water composition obtained in Example 1-3 has a significant blocking effect on the HepG2 cell cycle, with an increase in the G1 phase and a decrease in the S phase, which blocks the transition of cells from the G1 phase to the S phase, and has a good effect of inhibiting cancer cell proliferation.

[0150] Test Example 2 Effect on the Release of Inflammatory Factor NO

[0151] RAW264.7 cells were cultured in DMEM medium containing 10% fetal bovine serum and distilled water. When the cells reached the logarithmic growth phase, they were passaged or used in experiments. The cells were divided into a control group, a model group, Example 1-3 groups, and Comparative Example 1-10 groups. 4The cells were inoculated at a density of 100 cells / well in a 96-well culture plate. After the cells adhered to the wall, the original culture medium was discarded. The control group and the model group were added with 100 μL of culture medium prepared with distilled water, and the drug group was added with a culture medium prepared with a deuterium-depleted water composition (content of 10 wt%) for 2 hours. The model group and the drug group were induced with lipopolysaccharide solution (0.05 μg / mL) for 24 hours, 50 μL of the culture medium was drawn, 50 uL of Griess Reagent I and 50 uL of Griess Reagent Il were added, and the absorbance was measured at 540 nm. The results are shown in Table 3.

[0152] NO relative content (%) = absorbance of the drug administration group / absorbance of the control group × 100%

[0153] Table 3

[0154]

[0155]

[0156] It can be seen from the above table that the deuterium-depleted water compositions prepared in Examples 1-3 have an obvious effect of inhibiting the release of NO.

[0157] Test Example 4 Antioxidant Activity Test

[0158] Take 2 mL of the deuterium-depleted water composition prepared in Examples 1-3 or Comparative Examples 1-10, respectively, and put it into a stoppered graduated colorimetric tube, and add 0.6 mol / L sulfuric acid solution, 2.8 mmol / L Na3PO4 solution, 4 mmol / L (NH4)6Mo7O 24 2 mL of each solution was added with distilled water to make up to 10 mL, shaken and heated in a 95°C water bath for 150 min, taken out and cooled to room temperature, and the system without antioxidant was used as a blank reference, and the absorbance at a wavelength of 695 nm was measured, and a 1.0 mg / mL butylated hydroxytoluene ethanol solution was used as a positive control. The results are shown in Table 4.

[0159] Table 4

[0160]

[0161]

[0162] Under acidic conditions, the antioxidant components can convert Mo 6+ Reduction to Mo 5+ , phosphate ions react with Mo 5+A green phosphomolybdic acid complex is formed. The phosphomolybdic acid complex has strong absorption at 695nm. The antioxidant activity can be evaluated by comparing the absorbance of different deuterium-depleted water compositions after reacting with phosphomolybdic acid. The greater the absorbance, the stronger the reducing ability and the stronger the antioxidant property. As can be seen from the above table, the deuterium-depleted water compositions prepared in Examples 1-3 of the present invention have good antioxidant effects, which are better than the antioxidant butylated hydroxytoluene.

[0163] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a deuterium-depleted water composition, characterized in that: The following steps are involved: S1. Preparation of deuterium-depleted water: a hydrophobic catalyst is filled into an exchange column, preheated mineral water is introduced into the column from the top of the exchange column, and preheated high-purity hydrogen is introduced from the bottom of the exchange column. In the exchange column, a catalytic exchange reaction occurs between the high-purity hydrogen and water, and the product deuterium-depleted water is output from the bottom. The deuterium-depleted water is collected. After the production is completed, the exchange column is flushed and cleaned with high-purity nitrogen to regenerate the filler; S2. Extraction of active components of traditional Chinese medicine: Mix coptis root, poria, ginseng and Panax notoginseng, crush them, add low-deuterium water and heat to boiling for extraction, repeat 2-3 times, filter, retain the solid, combine the filtrate, concentrate, and dry to obtain the active components of traditional Chinese medicine; S3. Activation of bacteria: Selenium-enriched yeast and Lactobacillus plantarum were inoculated into Gao's medium, activated and cultured to obtain a culture medium with a bacterial content of 10 8 -10 9 cfu / mL of bacterial seed solution; S4. Deuterium-depleted water-assisted fermentation: adding the solid in step S2 to deuterium-depleted water, sterilizing, inoculating activated selenium-enriched yeast and Lactobacillus plantarum seed liquid, fermenting, filtering, concentrating the filtrate, and freeze-drying to obtain a fermentation component; S5. Preparation of deuterium-depleted water composition: Evenly mix the active components of traditional Chinese medicine, fermentation components and deuterium-depleted water to prepare the deuterium-depleted water composition.

2. The preparation method according to claim 1, characterized in that: The temperature of the preheated deionized water in step S1 is 40-60° C., the temperature of the preheated high-purity hydrogen is 40-60° C., the purity is >99.99%, the gas-liquid molar ratio of the deionized water and hydrogen is 0.4-0.7:1, the deuterium content of the low-deuterium water is ≤125ppm, and the half-peak width is <70Hz.

3. The preparation method according to claim 2, characterized in that: The preparation method of the hydrophobic catalyst is as follows: T1. Preparation of Pt / Pd@graphene: dissolve palladium chloride in hydrochloric acid to obtain H2PdCl4 solution, add chloroplatinic acid, stir and mix evenly, add graphene oxide, stir and react, dry, wash, pass hydrogen to heat and reduce, then pass hydrazine hydrate vapor to reduce, and obtain Pt / Pd@graphene; T2. Preparation of hydrophobic catalyst: dissolving fluorine-containing silane in an organic solvent to obtain an oil phase; adding a porogen, an emulsifier and Pt / Pd@graphene into water to obtain a suspension; dropping the suspension into the oil phase, emulsifying, adjusting the pH value of the solution, stirring the reaction, centrifuging, washing, spray drying, and obtaining a hydrophobic catalyst.

4. The preparation method according to claim 3, characterized in that: The concentration of the hydrochloric acid in step T1 is 0.3-0.5 mol / L, the mass ratio of the palladium chloride, chloroplatinic acid and graphene oxide is 3-5:7-12:30-50, the stirring reaction time is 12-15h, the temperature of the hydrogen heating reduction reaction is 300-350°C, the time is 2-4h, and the time of the hydrazine hydrate steam reduction is 3-5h.

5. The preparation method according to claim 3, characterized in that: The fluorine-containing silane in step T2 is selected from 1H, 1H, 2H, 2H-perfluorodecyltriethoxysilane, 1H, 1H, 2H, 2H-perfluorodecyltrimethoxysilane, dodecafluoroheptylpropyltrimethoxysilane, dodecafluoroheptylpropylmethyldimethoxysilane, 3,3,3-trifluoropropylmethyldimethoxysilane, 3,3,3-trifluoropropyltrimethoxysilane, 1H, 1H, 2H, 2H-perfluorooctyltriethoxysilane or 1H, 1H, 2H, 2H- At least one of perfluorooctyltrimethoxysilane, the mass ratio of the fluorinated silane, porogen, emulsifier and Pt / Pd@graphene is 12-15:0.5-1:0.2-0.4:6-8, the porogen is hexadecyltrimethylammonium chloride, the emulsifier is selected from at least one of Tween-20, Tween-40, Tween-60, Tween-80 and Tween-85, the pH value of the adjusted solution is 9-10, and the stirring reaction time is 5-7h.

6. The preparation method according to claim 1, characterized in that: In step S2, the mass ratio of coptis root, poria, ginseng, notoginseng and deuterium-depleted water is 3-7:2-4:1-2:3-5:100, and the time for heating to boiling extraction is 3-5h.

7. The preparation method according to claim 1, characterized in that: The conditions for the activation culture of the selenium-enriched yeast in step S3 are 45-50° C., 100-200 r / min, and activation culture for 18-24 h. The conditions for the activation culture of the plant lactobacillus are 35-40° C., 100-200 r / min, and activation culture for 18-24 h under anaerobic conditions.

8. The preparation method according to claim 1, characterized in that: The mass ratio of the solid to the deuterium-depleted water in step S4 is 10-15:100, the inoculation amounts of the activated selenium-enriched yeast and plant lactobacillus species seed liquid are 2-4v / v% and 1-2v / v%, respectively, the fermentation and culture conditions are under micro-anoxic conditions, 38-42°C, 100-200r / min, and the fermentation and culture is 36-48h, the micro-anoxic conditions are oxygen content 5-10v / v%, carbon dioxide content 3-7v / v%, and the balance is nitrogen; the mass ratio of the active components of the traditional Chinese medicine, the fermentation components and the deuterium-depleted water in step S5 is 2-3:1-2:

10.

9. A deuterium-depleted water composition obtained by the preparation method according to any one of claims 1 to 8.

10. Use of the deuterium-depleted water composition according to claim 9 in the preparation of anti-tumor, anti-inflammatory and antioxidant drugs.

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