Micro-molecular cluster water and application thereof in plant polysaccharide and plant nanometer polysaccharide

Abstract

The application provides micro-molecular cluster water and application thereof in plant polysaccharide and plant nanometer polysaccharide, and belongs to the technical field of nanometer materials. 17 The micro-molecular cluster water is prepared by using water phase liquid as raw material, sequentially performing adsorption through multiple-stage physical fillers and then performing activation filtration O-NMR half-peak width <100 Hz. The application uses the micro-molecular cluster water as solvent, utilizes complex enzymes to perform enzymolysis on plant materials, thereby obtaining plant polysaccharide liquid; then performs adsorption through macroporous adsorption resin, then performs elution by using the micro-molecular cluster water, performs decoloration on the eluate, performs high-pressure homogenization on the decolorized liquid, and then performs membrane filtration to obtain plant nanometer polysaccharide liquid. The plant nanometer polysaccharide liquid can significantly improve the stability of small-molecule water clusters, significantly inhibit the growth of melanin, and has a significant whitening effect.

Description

Technical Field

[0001] This invention belongs to the field of nanomaterials technology, specifically relating to a micro-molecular cluster water and its application in plant polysaccharides and plant nanopolysaccharides. Background Technology

[0002] Small water molecule clusters, due to their smaller molecular cluster structure, exhibit superior solubility, permeability, and bioactivity compared to ordinary water. They demonstrate enormous application potential in numerous fields, such as healthcare, food processing, and cosmetics manufacturing. However, small water molecule clusters possess a higher energy state and are prone to recombination into larger molecular clusters under natural conditions. Their stability remains a key factor limiting their widespread application. Polysaccharides are a class of biomolecules widely found in nature, possessing excellent biocompatibility, water solubility, and biodegradability. In recent years, nanoscale polysaccharides have attracted increasing attention across various fields due to their unique physicochemical properties, such as high specific surface area and quantum size effects. Plant polysaccharides exhibit a variety of biological activities, including antioxidant, anti-inflammatory, and immunomodulatory effects. Polysaccharide molecules are rich in polar functional groups such as hydroxyl (-OH) and carboxyl (-COOH) groups. These groups form strong hydrogen bonds with water molecules (H2O). Large clusters (typically containing more than 10 water molecules) formed by hydrogen bonds are competitively bound by the polar groups of polysaccharides, disrupting the stability of the original large hydrogen bond network. Nanoparticles (1-100 nm) of polysaccharides form a dispersed system in water. When some small water molecule clusters tend to aggregate due to molecular thermal motion, the polar groups of the surrounding nanoparticles quickly bind with free water molecules, reforming new small clusters. This dynamic regulation ensures that the proportion and size of small water molecule clusters remain stable throughout the system and do not significantly increase over time. However, there are currently no reports on the preparation of plant polysaccharides at the nanoscale and their application in stabilizing small water molecule clusters. Summary of the Invention

[0003] This invention provides a micro-molecular cluster water and its application in plant polysaccharides and plant nano-polysaccharides, which has the characteristics of high efficiency and environmental protection. Plant nano-polysaccharides can maintain the stability of the small molecule water cluster state, solve the problem of easy aggregation of small molecule water clusters, and expand the application scope of small molecule water clusters in various fields.

[0004] This invention provides micro-molecular cluster water, wherein the micro-molecular cluster water has... 17 O-NMR full width at half maximum (FWHM) < 100 Hz.

[0005] The present invention also provides a method for preparing the above-mentioned micro-molecular cluster water, comprising the following steps: after preliminary removal of impurities from the aqueous phase liquid, the following operations are performed in sequence: activation adsorption, reduction of molecular clusters, release of negative ions, reduction of the aqueous phase liquid, further removal of impurities and molecular cluster separation, to obtain the micro-molecular cluster water.

[0006] In one specific embodiment of the present invention, the preliminary impurity removal includes adsorption using activated carbon; the activation adsorption includes adsorption using porous Yunnan sand ceramic; the reduction of molecular clusters includes adsorption using far-infrared mineralized balls; the release of negative ions includes adsorption using tourmaline ceramic balls; the generation of reducing properties in the aqueous phase liquid includes adsorption using negative potential ceramic balls; the secondary impurity removal includes adsorption using maifanite mineralized balls and ion exchange resin; and the molecular cluster fragmentation includes adsorption using neodymium iron boron.

[0007] In one specific embodiment of the present invention, the water flow rate is 1~3 BV / h during the adsorption process.

[0008] This invention also provides the application of the above-mentioned micro-molecular cluster water or the micro-molecular cluster water prepared by the above preparation method in the extraction of polysaccharides from plant materials.

[0009] The present invention also provides a method for extracting plant polysaccharides, comprising the following steps: crushing plant material, mixing it with the above-mentioned micro-molecular cluster water or the micro-molecular cluster water prepared by the above preparation method and a complex enzyme, and then performing enzymatic hydrolysis, wherein the hydrolysate contains plant polysaccharides; The complex enzyme contains cellulase, hemicellulase, pectinase, alkaline protease, and amylase.

[0010] The present invention also provides a plant polysaccharide solution obtained by the above extraction method.

[0011] The present invention also provides a method for preparing plant nano polysaccharide solution, comprising the following steps: (1) after filtering the above plant polysaccharide solution, adsorb it using macroporous adsorption resin, and then elute it using the above micro molecular cluster water or the micro molecular cluster water prepared by the above preparation method. When the polysaccharide content of the collected eluent is 1.0 mg / mL to 12 mg / mL, the elution is stopped and a polysaccharide eluent is obtained. (2) Decolorize the polysaccharide solution obtained in step (1), homogenize the decolorized solution under high pressure, and then filter it with a membrane of less than 0.2 μm to obtain plant nano polysaccharide solution.

[0012] The present invention also provides a plant nanopolysaccharide liquid prepared using the above preparation method.

[0013] This invention also provides the application of the above-mentioned plant nanopolysaccharide liquid in medical and health care, food processing and / or cosmetic preparation.

[0014] Beneficial effects: This invention provides a micro-molecular cluster water, which is prepared by using an aqueous liquid as raw material, sequentially passing it through a multi-stage physical packing material for adsorption, followed by activation filtration. 17 Micro-molecular clusters of water with a half-peak width of O-NMR <100Hz.

[0015] This invention uses the aforementioned micro-molecular cluster water as a solvent and employs a composite enzyme to enzymatically hydrolyze plant materials, thereby obtaining a plant polysaccharide solution. This invention also uses the plant polysaccharide solution as a raw material, adsorbing it through a macroporous adsorption resin, then eluting it with the aforementioned micro-molecular cluster water, decolorizing the eluent, homogenizing the decolorized solution under high pressure, and finally filtering it through a membrane to obtain a plant nano-polysaccharide solution.

[0016] The plant-based polysaccharide nanoparticles prepared in this invention consist of sugar molecules linked by α-linkage bonds at positions 2, 3, and 6. These molecules interact with water molecules through weak hydrogen bonds and synergistic effects of water bridges, forming an ordered structure by excluding water molecules. These interactions create binding forces with water molecules, fixing water clusters around the polysaccharide molecules, without excessively restricting the movement of water molecules, thus maintaining small water clusters. The water bridging effect prevents the disintegration of individual water clusters and avoids the fusion of multiple water clusters into large water clusters.

[0017] In one embodiment of the present invention, a rose nanopolysaccharide solution prepared using rose as an example has been shown to significantly improve the stability of small molecule water clusters, significantly inhibit melanin growth, and have a significant whitening effect. Attached Figure Description

[0018] Figure 1 The condition of melanin in the head of a zebrafish; Figure 2 The image shows the intensity of melanin signal in zebrafish compared to the normal group. P <0.001、 P <0.01、 P <0.05; Figure 3 This is the initial half-width peak plot in Example 1; Figure 4 This is the initial half-width peak plot in Comparative Example 1; Figure 5 This is the initial half-width peak plot in Comparative Example 2; Figure 6 This is a half-width peak plot for 24 months (24M) in Example 1; Figure 7 This is a half-width broad peak plot for 24 months (24M) in Comparative Example 1; Figure 8 This is a half-width broad peak plot for 24 months (24M) in Comparative Example 2. Detailed Implementation

[0019] This invention provides micro-molecular cluster water, wherein the micro-molecular cluster water has... 17 O-NMR full width at half maximum (FWHM) < 100 Hz.

[0020] The present invention also provides a method for preparing the above-mentioned micro-molecular cluster water, comprising the following steps: after preliminary removal of impurities from the aqueous phase liquid, the following operations are performed in sequence: activation adsorption, reduction of molecular clusters, release of negative ions, reduction of the aqueous phase liquid, further removal of impurities and molecular cluster separation, to obtain the micro-molecular cluster water.

[0021] This invention uses an aqueous liquid as a raw material, which can be tap water, plant water, or purified water. The micro-molecular cluster water is obtained by treating the aqueous liquid. The treatment includes activation filtration using a packed column, where the column is filled with physical packing material, and the volume of the packed column is twice the volume of the aqueous liquid raw material being treated. In this embodiment of the invention, the aqueous liquid used is the juice obtained after breaking down the cell walls of the corresponding plant material.

[0022] The preliminary impurity removal of this invention includes adsorption using activated carbon; the activation adsorption includes adsorption using porous Yunnan sand ceramic; the reduction of molecular clusters includes adsorption using far-infrared mineralized balls; the release of negative ions includes adsorption using tourmaline ceramic balls; the reduction of water by using negative potential ceramic balls; the secondary impurity removal includes adsorption using maifanite mineralized balls and ion exchange resin; and the molecular cluster fragmentation includes adsorption using neodymium iron boron.

[0023] During the adsorption process of this invention, the water flow rate is 1~3 BV / h. In this invention, activated carbon is first used for preliminary impurity removal. The activated carbon has high porosity and ultra-large specific surface area, which can effectively adsorb organic impurities, pigments and odor molecules in the water, ensuring that the subsequent preparation process is not affected by impurities, thereby guaranteeing the final generation of micro-molecular cluster water.

[0024] This invention activates water by passing it through a packed column filled with porous Yunnan sandstone after adsorption by activated carbon. Yunnan sandstone is an iron-containing clayey siltstone composed of minerals such as hydromica, kaolinite, quartz, mica flakes, and iron. The main internal components of Yunnan sandstone are silicon dioxide and iron oxide, while the surface oxide layer has a nanoporous structure. After firing in a kiln, this oxide layer combines with the internal structure to activate water. The sintered surface forms a porous oxide layer with strong adsorption capacity, and the multi-layered structure enhances the adsorption capacity.

[0025] This invention utilizes porous Yunnan sand ceramic adsorption followed by a filling column with far-infrared mineralized spheres (also known as far-infrared negative ion spheres) as filler. This process reduces the molecular clusters of activated water and releases a large number of negative ions. The far-infrared mineralized spheres are functional water treatment filter media made by adding natural mineral raw materials such as maifanite and seabed sand to room-temperature infrared materials and firing them using ceramic technology. They release far-infrared rays of 4-14μm, breaking the hydrogen bonds between water molecules, converting large water molecule clusters into smaller ones, and simultaneously generating a pyroelectric effect, permanently releasing a large number of negative ions.

[0026] This invention involves adsorption by far-infrared mineralizing spheres, followed by passage through a packed column filled with tourmaline ceramic spheres (also known as tourmaline balls), thereby causing small water molecule clusters to release negative ions and hydroxyl radicals again. Tourmaline ceramic spheres are water treatment materials made by molding and sintering tourmaline as the core material with clay and other base materials. They possess permanent electrical polarity, forming an electric field on their surface that ionizes water and oxygen molecules, generating negative ions and hydroxyl radicals.

[0027] This invention involves adsorbing water using tourmaline ceramic balls, followed by passing the water through a packed column filled with negatively charged ceramic balls (also known as ORP reducing balls or antioxidant balls), thereby reducing the water's redox potential. The negatively charged ceramic balls are made from ultrafine mineral powders such as tourmaline, negative ion powder, and germanium powder through sintering, and can lower the redox potential of water. They release far-infrared energy, breaking the hydrogen bonds in water molecules and simultaneously generating a negative potential (-100~-500mV), thus giving the water reducing properties, increasing hydroxide ions in the water, and adjusting the pH to slightly alkaline.

[0028] This invention utilizes negatively charged ceramic balls for adsorption, followed by passage through a packed column filled with maifanite mineralized balls. This process effectively adsorbs heavy metals and organic matter from water while simultaneously regulating the water's pH level. The maifanite mineralized balls are a mineralized filter media made from a mixture of over 60% maifanite powder and clay, sintered at high temperatures. The maifanite balls exhibit a porous structure, a large specific surface area, and extremely strong adsorption capacity, enabling them to adsorb heavy metals and organic matter from water while also regulating the water's pH level.

[0029] This invention, after adsorption by maifanite mineralized spheres, further purifies and removes impurities from water of different sources and mineral ions that may have been introduced during the reaction process using a packed column filled with ion exchange resin. This ensures that the final product is standardized micro-molecular cluster water. Ion exchange resin is a high-molecular polymer particle with ion exchange function. Through the exchangeable ions on its surface, it undergoes a displacement reaction with ions in the water, specifically adsorbing heavy metals and removing Cl-. - Hypochlorite (ClO) - (and some organic impurities, salts, etc.)

[0030] This invention involves adsorption via ion exchange resin followed by passage through a packed column filled with neodymium iron boron (NdFeB). NdFeB is currently the most magnetically efficient permanent magnet material. By using NdFeB as the core magnetic source, ordinary water, in a specific magnetic field, flows perpendicularly through the magnetic field lines at a certain velocity. This alters the dipole moment orientation of water molecules, forming oriented chain or ring structures. Large water molecule clusters are broken down into smaller clusters. This process does not change the chemical composition of the water, but only its physical structure and properties.

[0031] This invention also provides the application of the above-mentioned micro-molecular cluster water or the micro-molecular cluster water prepared by the above preparation method in the extraction of polysaccharides from plant materials.

[0032] This invention does not specifically limit the type of plant material, which can be flowering plant material, rhizome plant material, stem and leaf plant material, or fruit material. Among them, flowering plant material can be rose, rhizome plant material can be Panax notoginseng, stem and leaf plant material can be Dendrobium, and fruit material can be coffee pulp.

[0033] The present invention also provides a method for extracting plant polysaccharides, comprising the following steps: crushing plant material, mixing it with the above-mentioned micro-molecular cluster water or the micro-molecular cluster water prepared by the above preparation method and a complex enzyme, and then performing enzymatic hydrolysis, wherein the hydrolysate contains plant polysaccharides; The complex enzyme contains cellulase, hemicellulase, pectinase, alkaline protease, and amylase.

[0034] This invention extracts plant polysaccharides using a compound enzyme hydrolysis method. Before extraction, the plant material is pulverized. The pulverized plant material is then mixed with the aforementioned micro-molecular cluster water and compound enzyme for enzymatic hydrolysis. The amount (v) of the micro-molecular cluster water is 2 to 10 times the amount (w) of the plant material (w) (w / v, unit: g / mL, kg / L), such as 2, 3, 4, 5, 6, 7, 8, 9, or 10 times. Then, the compound enzyme is added, with the amount (w) of the compound enzyme being 0.5 to 5% of the amount (w) of the plant material (w), such as 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, or 5%.

[0035] The composite enzyme of this invention includes cellulase, hemicellulase, pectinase, alkaline protease, and amylase. Pectinase breaks down the middle cell layer, cellulose, and hemicellulase disrupt the cell wall skeleton. Alkaline protease degrades glycoproteins and structural proteins to release polysaccharide binding, and amylase removes starch impurities and reduces system viscosity. The five enzymes work synergistically to deconstruct plant cell walls and intracellular matrix, efficiently disrupting the dense structure of plant tissues, promoting the full dissolution of bound and free polysaccharides, and reducing impurity interference, achieving the dual effect of increased extraction yield and improved polysaccharide purity. The mass ratio of cellulase, hemicellulase, pectinase, alkaline protease, and amylase in this invention is 2-5:1-3:1-3:1-3:2-5. All enzymes in the composite enzyme of this invention can be obtained commercially. In one embodiment, the cellulase activity is 10... 5 U / g, hemicellulase activity 2×10 5 U / g, pectinase activity 10 5 U / g, alkaline protease activity 2×10 5 U / g, α-amylase activity 10 5 U / g.

[0036] The enzymatic hydrolysis temperature described in this invention is 20~50℃, such as 20℃, 25℃, 30℃, 35℃, 40℃, 45℃ or 50℃, and the enzymatic hydrolysis time is 2~4h, such as 2h, 2.5h, 3h, 3.5h or 4h.

[0037] The present invention also provides a plant polysaccharide solution obtained by the above extraction method.

[0038] Using the extraction method described in this invention, the polysaccharides extracted from the plant polysaccharide solution are all sugar molecules linked by α-linkage bonds at positions 2, 3, and 6. These polysaccharides interact with water molecules through weak hydrogen bonds and synergistic effects of water bridges, forming an ordered structure by excluding water molecules. These interactions both bind to water molecules, fixing water clusters around the polysaccharide molecules, and do not excessively restrict the movement of water molecules, thus maintaining small water clusters. The water bridging effect prevents the disintegration of individual water clusters and avoids the fusion of multiple water clusters into large water clusters.

[0039] The present invention also provides a method for preparing plant nano polysaccharide solution, comprising the following steps: (1) after filtering the above plant polysaccharide solution, adsorb it using macroporous adsorption resin, and then elute it using the above micro molecular cluster water or the micro molecular cluster water prepared by the above preparation method. When the polysaccharide content of the collected eluent is 1.0 mg / mL to 12 mg / mL, the elution is stopped and a polysaccharide eluent is obtained. (2) Decolorize the polysaccharide solution obtained in step (1), homogenize the decolorized solution under high pressure, and then filter it with a membrane of less than 0.2 μm to obtain plant nano polysaccharide solution.

[0040] After extracting the plant polysaccharide solution, the present invention performs simple filtration through a sieve, such as filtering through a 100-mesh sieve in the example, to obtain the filtered extract. The filtered extract is then passed through a macroporous adsorption resin column for adsorption. The macroporous adsorption resin can be one or more of AB-8, XAD-8, and HP20.

[0041] After the adsorption process, the present invention uses the aforementioned micro-molecular cluster water for elution at a flow rate of 1-3 BV / h. The eluent is collected from the point of clarity until the polysaccharide content in the collected eluent is 1.0 mg / mL to 12 mg / mL. Elution is then stopped to obtain the polysaccharide eluent.

[0042] This invention decolorizes the polysaccharide eluent by activated carbon adsorption. The amount (w) of activated carbon used during adsorption is 1-10% of the amount (w) of the plant material, such as 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%. After mixing the activated carbon with the polysaccharide eluent and stirring evenly for 30-50 minutes, the mixture is passed through a tilting filter to remove the activated carbon, and the decolorized liquid is collected.

[0043] This invention involves subjecting the decolorizing solution to high-pressure homogenization. The homogenization pressure is 80-150 MPa, such as 80 MPa, 90 MPa, 100 MPa, 110 MPa, 120 MPa, 130 MPa, 140 MPa, or 150 MPa; the homogenization temperature is 10-40℃, such as 10℃, 15℃, 20℃, 25℃, 30℃, 35℃, or 40℃. After 2-5 cycles of high-pressure homogenization, the solution is filtered through a precision membrane with a diameter of less than 0.2 μm to obtain a plant nano-polysaccharide solution.

[0044] The present invention also provides a plant nanopolysaccharide liquid prepared using the above preparation method.

[0045] Using the preparation method described in this invention, rose nanopolysaccharide solution, coffee pericarp nanopolysaccharide solution, Panax notoginseng nanopolysaccharide solution and Dendrobium nanopolysaccharide solution can be prepared; and the various plant nanopolysaccharide solutions prepared have good stability.

[0046] This invention also provides the application of the above-mentioned plant nanopolysaccharide liquid in medical and health care, food processing and / or cosmetic preparation.

[0047] One embodiment of the present invention is illustrated using rose nanopolysaccharide liquid as an example, which has a significant whitening effect.

[0048] The exemplary numbers given in this specification are for illustrative purposes only, and may also be any numbers between any two numbers in the form of integers, decimals, or fractions.

[0049] To further illustrate the present invention, the following detailed description, in conjunction with embodiments, of a micro-molecular cluster water provided by the present invention and its application in plant polysaccharides and plant nano-polysaccharides, should not be construed as limiting the scope of protection of the present invention.

[0050] Example 1: Preparation of Rose Nanoparticle Polysaccharide Solution (1) Preparation of micro-molecular cluster water: Take 50 kg of fresh rose petals, break them down to release the rose juice, then press and filter through a 100-mesh fine sieve to obtain rose juice. Then, pass the rose juice through a packed column containing physical fillers such as activated carbon, porous Yunnan sand ceramic, far-infrared mineralized balls, tourmaline ceramic balls, negative potential ceramic balls, maifanite mineralized balls, ion exchange resin, and neodymium iron boron at a flow rate of 2 BV / h for activation filtration to prepare... 17 Micro-molecular clusters of water with a half-peak width of O-NMR <100Hz are available for use.

[0051] (2) Polysaccharide extraction: Take another 2 kg of fresh rose petals and crush them. Add 10 L (5 times the amount, w / v) of the micro-molecular cluster water obtained in step (1), add 50 g (2.5% of the rose petal mass, w / w) of complex enzyme, and enzymatically hydrolyze at 35℃ for 3 h. The complex enzyme is compounded in a mass ratio of cellulase: hemicellulase: pectinase: alkaline protease: amylase of 3:1:1:1:2. Among them, the cellulase activity is 10 5 U / g, hemicellulase activity 2×10 5 U / g, pectinase activity 10 5 U / g, alkaline protease activity 2×10 5 U / g, α-amylase activity 10 5 U / g.

[0052] (3) Polysaccharide separation: After extraction, the extract was released and filtered through a 100-mesh sieve to obtain the filtered extract. The extract was then passed through three macroporous adsorption resin columns, namely AB-8, XAD-8, and HP20, for adsorption. The column volume of each resin was 2L. After adsorption, impurities were first eluted with water containing micro-molecular clusters obtained in step (1) at a flow rate of 2 BV / h. The eluent was collected from the point of clarity until the polysaccharide content in the collected eluent was 1.0 mg / mL to 12 mg / mL. Elution was then stopped, and the eluents were combined to obtain the polysaccharide eluent.

[0053] (4) Decolorization: Take out the eluent obtained in step (3), add 100g of activated carbon (5% of the rose's mass, w / w), stir evenly for 40min, then pass it through a tilting filter to remove the activated carbon and collect the decolorized liquid.

[0054] (5) The decolorizing solution is subjected to high-pressure homogenization at a pressure of 100 MPa and a homogenization temperature of 20°C. After three cycles, it is filtered through a precision membrane of less than 0.2 μm to obtain rose nanopolysaccharide solution.

[0055] Example 2: Preparation of coffee fruit peel nanopolysaccharide solution (1) Preparation of micro-molecular cluster water: Take 50 kg of fresh coffee pulp, break it down to release the juice, then press it and filter it through a 100-mesh sieve to obtain coffee juice. Then, pass the coffee juice through a packed column containing physical fillers such as activated carbon, porous Yunnan sand ceramic, far-infrared mineralized balls, tourmaline ceramic balls, negative potential ceramic balls, maifanite mineralized balls, ion exchange resin, and neodymium iron boron at a flow rate of 2 BV / h for activation filtration to prepare... 17 Micro-molecular clusters of water with a half-peak width of O-NMR <100Hz are available for use.

[0056] (2) Polysaccharide extraction: Take another 2 kg of fresh coffee pulp and crush it. Add 16 L (8 times the amount, w / v) of the micro-molecular cluster water obtained in step (1), add 10 g (0.5% of the coffee pulp mass, w / w) of complex enzyme, and enzymatically hydrolyze at 50℃ for 4 h. The complex enzyme is compounded in the mass ratio of cellulase: hemicellulase: pectinase: alkaline protease: amylase 2:1:3:2:3. Among them, the cellulase activity is 10 5 U / g, hemicellulase activity 2×10 5 U / g, pectinase activity 10 5 U / g, alkaline protease activity 2×10 5 U / g, α-amylase activity 10 5 U / g.

[0057] (3) Polysaccharide separation: After extraction, the extract was released and filtered through a 100-mesh sieve to obtain the filtered extract. The extract was then passed through three macroporous adsorption resin columns, namely AB-8, XAD-8, and HP20, for adsorption. The column volume of each resin was 2L. After adsorption, impurities were first eluted with water containing micro-molecular clusters obtained in step (1) at a flow rate of 1 BV / h. The eluent was collected from the point of clarity until the polysaccharide content in the collected eluent was 1.0 mg / mL to 12 mg / mL. Elution was then stopped, and the eluents were combined to obtain the polysaccharide eluent.

[0058] (4) Decolorization: Take out the eluent obtained in step (3), add 200g of activated carbon (10% of the coffee pulp mass, w / w), stir evenly for 50min, then pass it through a tilting filter to remove the activated carbon and collect the decolorized liquid.

[0059] (5) The decolorizing liquid is subjected to high pressure homogenization treatment at a pressure of 150 MPa and a homogenization temperature of 10°C. After two cycles, it is filtered with a precision membrane of less than 0.2 μm to obtain coffee pulp nano polysaccharide liquid.

[0060] Example 3: Preparation of Panax notoginseng nanopolysaccharide solution (1) Preparation of micro-molecular cluster water: Take 50 kg of fresh Panax notoginseng flower and leaf, break down the cell wall and crush it, then press it, and filter it through a 100-mesh fine sieve to obtain Panax notoginseng flower and leaf juice. Then, pass the Panax notoginseng flower and leaf juice through a packed column containing physical fillers such as activated carbon, porous Yunnan sand ceramic, far-infrared mineralized balls, tourmaline ceramic balls, negative potential ceramic balls, maifanite mineralized balls, ion exchange resin, and neodymium iron boron at a flow rate of 1 BV / h for activation filtration to prepare 17 Micro-molecular clusters of water with a half-peak width of O-NMR <100Hz are available for use.

[0061] (2) Polysaccharide extraction: Take 2 kg of fresh Panax notoginseng roots and crush them. Add 4 L (2 times the amount, w / v) of the micro-molecular cluster water obtained in step (1), add 100 g (5% of the mass of Panax notoginseng roots, w / w) of complex enzyme, and enzymatically hydrolyze at 20℃ for 4 h. The complex enzyme is compounded in the mass ratio of cellulase: hemicellulase: pectinase: alkaline protease: amylase 5:3:1:3:5. Among them, the cellulase activity is 10 5 U / g, hemicellulase activity 2×10 5 U / g, pectinase activity 10 5 U / g, alkaline protease activity 2×10 5 U / g, α-amylase activity 10 5 U / g.

[0062] (3) Polysaccharide separation: After extraction, the extract was released and filtered through a 100-mesh sieve to obtain the filtered extract. The extract was then passed through three macroporous adsorption resin columns, namely AB-8, XAD-8, and HP20, for adsorption. The column volume of each resin was 2L. After adsorption, impurities were first eluted with water containing micro-molecular clusters obtained in step (1) at a flow rate of 2 BV / h. The eluent was collected from the point of clarity until the polysaccharide content in the collected eluent was 1.0 mg / mL to 12 mg / mL. Elution was then stopped, and the eluents were combined to obtain the polysaccharide eluent.

[0063] (4) Decolorization: Take out the eluent obtained in step (3), add 20g of activated carbon (1% of the mass of Panax notoginseng root, w / w), stir evenly for 40min, then pass it through a tilting filter to remove the activated carbon and collect the decolorized liquid.

[0064] (5) The decolorizing solution is subjected to high pressure homogenization treatment at a pressure of 80 MPa and a homogenization temperature of 40 °C. After 5 cycles, it is filtered with a precision membrane of less than 0.2 μm to obtain Panax notoginseng nano polysaccharide solution.

[0065] Example 4: Preparation of Dendrobium nanopolysaccharide solution (1) Preparation of micro-molecular cluster water: Take 50 kg of fresh Dendrobium stems and leaves, break down the cell walls and crush them to release the Dendrobium stem and leaf juice, then press it and filter it through a 100-mesh fine sieve to obtain Dendrobium stem and leaf juice. Then, pass the Dendrobium stem and leaf juice through a packed column containing physical fillers such as activated carbon, porous Yunnan sand ceramic, far-infrared mineralized balls, tourmaline ceramic balls, negative potential ceramic balls, maifanite mineralized balls, ion exchange resin, and neodymium iron boron at a flow rate of 3 BV / h for activation filtration to prepare the following water: 17 Micro-molecular clusters of water with a half-peak width of O-NMR <100Hz are available for use.

[0066] (2) Polysaccharide extraction: Take 2 kg of fresh Dendrobium stems and crush them. Add 20 L (10 times the amount, w / v) of the micro-molecular cluster water obtained in step (1), add 20 g (1% of the Dendrobium stem mass, w / w) of complex enzyme, and enzymatically hydrolyze at 40℃ for 2 h. The complex enzyme is compounded in a mass ratio of cellulase: hemicellulase: pectinase: alkaline protease: amylase of 4:2:1:3:2. Among them, the cellulase activity is 10 5 U / g, hemicellulase activity 2×10 5 U / g, pectinase activity 10 5 U / g, alkaline protease activity 2×10 5 U / g, α-amylase activity 10 5 U / g.

[0067] (3) Polysaccharide separation: After extraction, the extract was released and filtered through a 100-mesh sieve to obtain the filtered extract. The extract was then passed through three macroporous adsorption resin columns, namely AB-8, XAD-8, and HP20, for adsorption. The column volume of each resin was 2L. After adsorption, impurities were first eluted with water containing micro-molecular clusters obtained in step (1) at a flow rate of 3 BV / h. The eluent was collected from the point of clarity until the polysaccharide content in the collected eluent was 1.0 mg / mL to 12 mg / mL. Elution was then stopped, and the eluents were combined to obtain the polysaccharide eluent.

[0068] (4) Decolorization: Take out the eluent obtained in step (3), add 50g of activated carbon (2.5% of the mass of Dendrobium stem strips, w / w), stir evenly for 30min, then pass it through a tilting filter to remove the activated carbon and collect the decolorized liquid.

[0069] (5) The decolorizing solution is subjected to high pressure homogenization treatment at a pressure of 120 MPa and a homogenization temperature of 30°C. After two cycles, it is filtered with a precision membrane of less than 0.2 μm to obtain Dendrobium nanopolysaccharide solution.

[0070] Comparative Example 1: Preparation of plant-derived micro-molecular cluster water 50 kg of fresh rose petals were crushed to release the rose juice, which was then pressed and filtered through a 100-mesh sieve to obtain rose juice. The rose juice was then passed sequentially at a flow rate of 2 BV / h through a packed column containing activated carbon, porous Yunnan sand ceramic, far-infrared mineralized balls, tourmaline ceramic balls, negative potential ceramic balls, maifanite mineralized balls, ion exchange resin, and NdFeB resin for activation filtration, thus preparing... 17 Micro-molecular clusters of water with a half-peak width of O-NMR <100Hz are available for use.

[0071] Comparative Example 2: Preparation of ordinary micro-molecular cluster water Take 50 kg of purified water and pass it sequentially through a packed column containing physical fillers such as activated carbon, porous Yunnan sand ceramic, far-infrared mineralized balls, tourmaline ceramic balls, negative potential ceramic balls, maifanite mineralized balls, ion exchange resin, and neodymium iron boron at a flow rate of 2 BV / h for activation filtration, and prepare the following product: 17 Micro-molecular clusters of water with a half-peak width of O-NMR <100Hz are available for use.

[0072] Experimental Example 1: Water Stability Test of Micromolecular Clusters Samples from Examples 1, 2, 3, 4, Comparative Example 1, and Comparative Example 2 were placed in an environment with a temperature of 25°C and a relative humidity of 50%. The half-width of water molecule clusters was measured using NMR technology every 6 months (6M) until 24 months. The method for measuring the half-width of water molecule clusters was based on the standard "JY / T 0578-2020 General Rules for Testing Methods of Superconducting Pulse Fourier Transform Nuclear Magnetic Resonance Spectroscopy".

[0073] Table 1 Stability Test Results

[0074] The results are shown in Table 1. During the 24-month stability test, the half-width of the micro-cluster water in Example 1 (containing rose nanopolysaccharides) decreased from 52.48 Hz. Figure 3 The frequency gradually increased to 89.50 Hz. Figure 6 The increase was only 70.6%, while the comparative ratio 1 ( Figure 4 and Figure 7 ) and Comparative Example 2 ( Figure 5 and Figure 8 The Hz and Hz values ​​increased to 106.67 Hz and 116.53 Hz respectively, with increases exceeding 100%. This indicates that rose nanopolysaccharides significantly improved the stability of small water molecule clusters.

[0075] Experimental Example 2: Application in Cosmetics 1. Principles and methods: The primary mechanism by which skincare products achieve whitening effects is by inhibiting the activity of tyrosinase, thereby reducing melanin production. The melanin regulation mechanism in zebrafish skin is highly conserved with that in humans, and the rate of melanin production is rapid, making it clearly observable under a microscope. Image processing technology can be used to quantify the degree to which skincare products reduce melanin, thus enabling the evaluation of whitening efficacy.

[0076] 2. System and sample size: (1) Zebrafish strain: Wild-type AB strain zebrafish.

[0077] (2) Age of zebrafish: 6 hours after fertilization (hpf).

[0078] (3) Sample size per group: 15 tails.

[0079] (4) Adult fish rearing and breeding methods: Refer to DB43 / T 1022-2015 Technical Specifications for Indoor Culture of Zebrafish.

[0080] 3. Reagents, consumables, and instruments: (1) Reagents and consumables: water, deionized water or water of equivalent purity; arbutin; sodium bicarbonate; potassium chloride; calcium chloride dihydrate; magnesium sulfate heptahydrate; 6-well microplate; (2) Instruments: stereomicroscope; electronic balance; constant temperature incubator; and other conventional laboratory instruments and equipment, such as pipettes, volumetric flasks, beakers, aluminum foil, pipettes, etc.

[0081] 4. Experimental steps: (1) Select 6hpf wild-type AB strain zebrafish in a 6-well plate.

[0082] (2) No treatment was given to the normal group, and 0.01% (m / m) arbutin was given to the positive group. The test sample group was treated with 1% (m / m) polysaccharide solution prepared in Examples 1 to 4, 1% (m / m) micro molecular rose cluster water prepared in Comparative Example 1, and 1% (m / m) micro molecular cluster water prepared in Comparative Example 2. The samples were incubated at 28°C for 45 h.

[0083] (3) Ten zebrafish were randomly selected from each group, and their heads were photographed under a dissecting microscope and the images were saved.

[0084] (4) The intensity of melanin signal in the head of zebrafish was analyzed using Image J (S, the parameter was calibrated to OD value during Image J software analysis); the statistical results were expressed as mean ± SE; Dunnett's T-test was used for statistical analysis, and the melanin signal intensity of each experimental group was compared with the normal control group as the standard. p<0.05 was considered to be statistically significant.

[0085] The formula for calculating the inhibitory effect of the test sample on melanin formation is as follows: 5. Judgment basis: (1) Compared with the normal control group, the melanin in the heads of zebrafish in the test group was significantly reduced, proving that the sample has a whitening effect.

[0086] (2) Has efficacy ( P <0.05 is considered statistically significant.

[0087] The effects of each group of samples on melanin production are as follows: Figure 1 As shown, compared with the normal control group, the melanin in the heads of zebrafish was reduced in 1% of Example 1, 1% of Comparative Example 1, 1% of Example 2, 1% of Example 3, and 1% of Example 4, revealing that Example 1, Example 2, Example 3, Example 4, and Comparative Example 1 have whitening effects.

[0088] Table 2. Inhibitory effect of test samples on melanin formation

[0089] Statistical analysis was performed on the melanin formation in each group, such as... Figure 2 As shown in Table 2, compared with the normal group, arbutin and those from Examples 1, 2, 3, 4, and Comparative Example 1 all significantly inhibited melanin formation. P <0.001), indicating that the samples of Example 1, Example 2, Example 3, Example 4 and Comparative Example 1 all have whitening effects.

[0090] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A type of micro-molecular cluster water, characterized in that, The micro-molecular clusters of water 17 O-NMR full width at half maximum (FWHM) < 100 Hz.

2. The method for preparing micro-molecular cluster water according to claim 1, characterized in that, The process includes the following steps: after preliminary impurity removal from the aqueous phase liquid, the following operations are performed in sequence: activation adsorption, reduction of molecular clusters, release of negative ions, reduction of the aqueous phase liquid to produce reducing properties, further impurity removal and molecular cluster separation, to obtain the micro molecular cluster water.

3. The preparation method according to claim 2, characterized in that, The initial impurity removal includes adsorption using activated carbon; the activation adsorption includes adsorption using porous Yunnan sand ceramic; the reduction of molecular clusters includes adsorption using far-infrared mineralized balls; the release of negative ions includes adsorption using tourmaline ceramic balls; the reduction of the aqueous phase liquid includes adsorption using negative potential ceramic balls; the secondary impurity removal includes adsorption using maifanite mineralized balls and ion exchange resin; and the molecular cluster fragmentation includes adsorption using neodymium iron boron.

4. The preparation method according to claim 3, characterized in that, During the adsorption process, the flow rate of the aqueous phase liquid is 1~3 BV / h.

5. The application of the micro-molecular cluster water of claim 1 or the micro-molecular cluster water prepared by the preparation methods of claims 2 to 4 in the extraction of polysaccharides from plant materials.

6. A method for extracting plant polysaccharides, characterized in that, Includes the following steps: After the plant material is crushed, it is mixed with the micro-molecular cluster water described in claim 1 or the micro-molecular cluster water prepared by the preparation method described in claims 2 to 4 and the complex enzyme, and then enzymatically hydrolyzed. The enzymatic hydrolysate contains plant polysaccharides. The complex enzyme contains cellulase, hemicellulase, pectinase, alkaline protease, and amylase.

7. The plant polysaccharide solution obtained by the extraction method described in claim 6.

8. A method for preparing a plant nanopolysaccharide solution, characterized in that, The process includes the following steps: (1) After filtering the plant polysaccharide solution described in claim 7, adsorb it using a macroporous adsorption resin, and then elute it using the micro-molecular cluster water described in claim 1 or the micro-molecular cluster water prepared by the preparation methods described in claims 2 to 4. When the polysaccharide content of the collected eluent is 1.0 mg / mL to 12 mg / mL, stop elution and obtain the polysaccharide eluent. (2) Decolorize the polysaccharide solution obtained in step (1), homogenize the decolorized solution under high pressure, and then filter it with a membrane of less than 0.2 μm to obtain plant nano polysaccharide solution.

9. The plant nanopolysaccharide liquid prepared by the preparation method according to claim 8.

10. The application of the plant nanopolysaccharide liquid of claim 9 in medical and health care, food processing and / or cosmetic preparation.

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