A low-deuterium water-based anthocyanin composite stabilizing system and its application in multi-category food and beverage

Abstract

The application belongs to the technical field of health-care food, and particularly relates to a low-deuterium water-based anthocyanin composite stabilizing system and application of the system to multiple types of food and drink, wherein the low-deuterium water-based anthocyanin composite stabilizing system comprises a triple synergistic stabilizing hierarchy: a beta-cyclodextrin molecular level embedding, a liposome biomimetic wrapping layer, and a low-deuterium water carrier environment, and the three synergistically greatly improve the time retention rate of anthocyanin and the cell absorption rate, and the bioavailability is high. Based on the core system, the application further develops low-deuterium water anthocyanin beverages, synergistic antioxidant compositions, three-high health-care tea, stomach-nourishing rice paste, anthocyanin health-care fresh beer and health-care liquor and other products of different types, realizes cross-class application of the same stable anthocyanin product food, drink and liquor, and has the characteristics of high retention rate, good bioavailability, green environmental protection and industrialization feasibility.

Description

Technical Field

[0001] This invention belongs to the field of health and wellness food technology, specifically relating to a low-deuterium water-based anthocyanin complex stabilization system and its application in various types of food and beverages. Background Technology

[0002] Anthocyanins are water-soluble flavonoid pigments widely found in nature, derived from plants such as black goji berries, blueberries, mulberries, and purple sweet potatoes. They possess a variety of biological activities, including strong antioxidant, anti-inflammatory, vision-improving, and cardiovascular disease-preventing effects. Their DPPH free radical scavenging IC50 is only 8–15 μg / mL, and their antioxidant capacity is 20–50 times that of vitamin C.

[0003] Anthocyanins can effectively enhance the nutritional value and functional properties of food. However, their application in the food industry faces the following challenges: 1. Poor stability: Anthocyanins are highly sensitive to temperature, light, pH, and oxygen. Under normal temperature conditions, the retention rate after 30 days is typically less than 50%, severely limiting product shelf life. 2. Low bioavailability: After oral administration, anthocyanins undergo gastrointestinal metabolism and molecular degradation, resulting in an actual absorption rate of only 20%–30%, far lower than in vitro activity test results. 3. Limited carrier technology: Existing technologies mostly employ single methods such as β-cyclodextrin encapsulation or liposome encapsulation, offering limited protection. The retention rate at room temperature generally does not exceed 65%, and the synergistic potential between various carriers has not been fully explored. 4. Lack of unified products: Different categories of anthocyanin products are developed independently, leading to redundant R&D investment and a lack of core stable products that can be transferred across product categories. Summary of the Invention

[0004] The present invention aims to provide a low-deuterium water-based anthocyanin complex stabilization system and its application in multiple categories of food and beverages. This low-deuterium water-based anthocyanin complex stabilization system enables the cross-category application of anthocyanin products, achieves high retention rate, high bioavailability and long-term stability of anthocyanins, and reduces the development cost of anthocyanin products.

[0005] Deuterium-depleted water (DDW) refers to water with a deuterium content lower than that of ordinary natural water (approximately 150 ppm), typically ranging from 30 to 120 ppm. Studies have shown that DDW possesses the following characteristics: smaller molecular clusters, with approximately 3-5 water molecules clustered together (compared to 10-15 in ordinary water), resulting in stronger cell membrane penetration; it can increase the activity of intracellular antioxidant enzymes such as SOD and CAT by 15%-20%; and the CD bond energy (363 kJ / mol) formed by deuterium replacing hydrogen is higher than that of the CH bond (347 kJ / mol), resulting in a more stable molecular skeleton. Currently, there are no reports of using DDW as an anthocyanin carrier in conjunction with cyclodextrin encapsulation and liposome encapsulation to form a triple synergistic stabilizing system. This invention, based on the characteristics of anthocyanins and combined with the functions of DDW, develops a unified product that can significantly improve the stability and bioavailability of anthocyanins and can be applied across multiple product categories.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: A method for preparing a low-deuterium water-based anthocyanin complex stable system is provided, comprising the following steps: S1: Obtain an anthocyanin-containing extract; S2: The anthocyanin extract from step S1 was encapsulated with β-cyclodextrin in deuterated water to obtain an anthocyanin-β-cyclodextrin encapsulated complex. S3: Dissolve phospholipids in anhydrous ethanol, dry them with nitrogen to form a film, hydrate them with deuterium-free water, and homogenize them by ultrasound or high pressure to obtain a liposome suspension. S4: Disperse the encapsulated complex from step S2 in the liposome suspension from step S3, and obtain a low-deuterium water-based anthocyanin complex stable system after lyophilization or sterile filtration.

[0007] Preferably, in step S1, the method for obtaining the anthocyanin-containing extract is as follows: Using at least one of black goji berries, purple sweet potatoes, blueberries, and mulberries as raw materials, anthocyanins are extracted and purified by acidic ethanol to obtain freeze-dried powder with an anthocyanin content ≥80%; or, Using at least one of black goji berries, purple sweet potatoes, blueberries, and mulberries as raw materials, and using low-deuterium water with a deuterium content ≤80ppm instead of acidic ethanol as the extraction solvent, the anthocyanin raw materials are subjected to ultrasonic-assisted extraction to obtain anthocyanin extract, which is then used in step S2 to directly add β-cyclodextrin to the anthocyanin extract for in-situ encapsulation.

[0008] Preferably, in step S2, the molar ratio of anthocyanin to β-cyclodextrin in the obtained anthocyanin-β-cyclodextrin encapsulated complex is 1:2 to 1:3, and the encapsulation rate is ≥85%; in step S3, the liposomes in the liposome suspension have a particle size of 80 to 200 nm and a polydispersity index ≤0.3; and the deuterium content of the low-deuterium water in steps S2 and S3 is ≤50 ppm.

[0009] Preferably, in step S2, the embedding temperature is 45±2℃, the embedding time is 1.5~2.5h, and the embedding reaction is carried out by stirring at a speed of 180~220rpm. In step S3, the phospholipid is soybean lecithin or hydrogenated soybean lecithin, and the zeta potential of the liposome is -30 to -50 mV; the high-pressure homogenization conditions in step S3 are 95 to 105 MPa and 4 to 6 cycles.

[0010] In step S4, the embedding complex and liposome suspension are added in a mass ratio of 9:1 to 11:1 between the liposome suspension and the embedding complex.

[0011] The present invention also provides a low-deuterium water-based anthocyanin complex stabilization system prepared by the method described above for preparing a low-deuterium water-based anthocyanin complex stabilization system.

[0012] The present invention also provides a low-deuterium water-based anthocyanin beverage, comprising the aforementioned low-deuterium water-based anthocyanin complex stabilization system, using low-deuterium water with a deuterium content ≤50ppm as the matrix, the addition amount of the low-deuterium water-based anthocyanin complex stabilization system is 500~2000g / 1000L, the pH value is 3.8~4.2, nitrogen filling is used and the headspace oxygen is ≤1%.

[0013] The present invention also provides a low-deuterium water-anthocyanin-Polygonatum polysaccharide synergistic antioxidant composition, comprising the aforementioned low-deuterium water-based anthocyanin complex stabilizing system and Polygonatum polysaccharide extract, wherein the polysaccharide content in the Polygonatum polysaccharide extract is ≥50%, and the mass ratio of the low-deuterium water-based anthocyanin complex stabilizing system to the Polygonatum polysaccharide extract is 1:1 to 1:5.

[0014] The present invention also provides a low-deuterium water-based health tea with synergistic regulation of high blood pressure, high blood sugar, and high cholesterol, comprising the aforementioned low-deuterium water-based anthocyanin complex stabilizing system, as well as black goji berry freeze-dried powder, polygonatum ultrasonic extraction concentrate, mulberry enzymatic fermentation broth, hawthorn supercritical CO2 extract and cassia seed water-extracted alcohol precipitate. The components are compounded in a gradient manner under ≤4℃ and inert gas protection.

[0015] The present invention also provides a low-deuterium water-based stomach-nourishing rice paste, comprising the aforementioned low-deuterium water-based anthocyanin complex stabilizing system, and a complex of millet, black rice, monkey head mushroom, yam, polygonatum, black goji berries, mulberry, red dates, oats, and milk powder or yogurt, wherein the monkey head mushroom is pulverized by airflow to a D90≤10μm; the rice paste is a powder-based reconstituted type or a bottled ready-to-drink type.

[0016] The present invention also provides a low-deuterium water-based anthocyanin health-preserving fresh beer, comprising the aforementioned low-deuterium water-based anthocyanin complex stabilizing system, using low-deuterium water with a deuterium content ≤50ppm as brewing water to brew the beer, and adding the aforementioned low-deuterium water-based anthocyanin complex stabilizing system as a modifier to the beer after filtration, with an alcohol content of 4-6% vol, and using nitrogen filling or micro-freezing preservation bottling.

[0017] This invention also provides a low-deuterium water-based anthocyanin health-preserving baijiu, comprising the aforementioned low-deuterium water-based anthocyanin complex stabilization system. It uses a medium-low alcohol baijiu (20-38% vol) as the base spirit, and after being diluted and blended with low-deuterium water, it is processed through anthocyanin-Zn... 2+ The complex and the low-deuterium water-based anthocyanin complex stabilization system were modified and then bottled and stored.

[0018] Compared with the prior art, the beneficial effects of the present invention are: (1) The low-deuterium water-based anthocyanin composite stabilization system has triple synergistic stability and significant effect. Among them, the β-cyclodextrin encapsulation, liposome encapsulation and low-deuterium water environment work together to significantly improve the retention rate of anthocyanins over time. (2) The low-deuterium water-based anthocyanin composite stabilization system can be applied to different categories such as beverages, antioxidant compositions, health teas, rice paste, fresh beer, and liquor, covering the three major industries of food, beverages and alcohol, avoiding repeated R&D investment. (3) The synergistic antioxidant effect is prominent. Low-deuterium water can activate endogenous antioxidant enzymes, and anthocyanins can directly scavenge free radicals. The antioxidant pathway is synergistic, and the comprehensive antioxidant coefficient is improved. (4) Using low-deuterium water instead of acidic ethanol as the extraction solvent, the anthocyanin yield is improved compared with the traditional acid-ethanol method, and there is no organic solvent residue. After extraction, it can be directly encapsulated in situ, simplifying the process. The low-deuterium water carrier improves the cell absorption rate of anthocyanins and enhances bioavailability. Detailed Implementation

[0019] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0020] Example 1: A method for preparing a low-deuterium water-based anthocyanin complex stabilization system The method for preparing a low-deuterium water-based anthocyanin complex stabilization system includes the following steps: S1: Obtain an anthocyanin-containing extract.

[0021] Using black goji berries as raw material, anthocyanins were extracted by crushing and then extracted with acidic ethanol (0.1% HCl, pH 3.0, 60% ethanol) at 45℃ for 1 hour. The anthocyanins were then purified by adsorption with AB-8 macroporous resin and elution with 70% ethanol. The collected eluent was freeze-dried (-50℃, 0.1 mbar) to obtain anthocyanin freeze-dried powder with an anthocyanin content ≥80% (HPLC method, cyanidin-3-O-glucoside standard).

[0022] S2: The anthocyanin extract from step S1 is encapsulated with β-cyclodextrin in deuterated water to obtain an anthocyanin-β-cyclodextrin encapsulated complex.

[0023] In step S2, β-CD (β-cyclodextrin) is first dissolved in deuterated water (5% w / v, dissolved at 60℃, deuterium content of deuterated water ≤50ppm), and then the anthocyanin lyophilized powder prepared in step 1 (anthocyanin:β-CD=1:2.5 molar ratio) is added, and the mixture is stirred and embedded (45℃ / 2h / 200rpm). The embedded complex is then freeze-dried to obtain the embedded complex (-50℃, 0.1mbar, 24h), with an embedding rate ≥85% (determined by UV-Vis method).

[0024] S3: The phospholipids were dissolved in anhydrous ethanol, dried with nitrogen to form a film, hydrated with deuterium-free water, and then sonicated to obtain a liposome suspension.

[0025] In step S3, the phospholipid is soybean lecithin, which is dissolved in anhydrous ethanol (50 mg / ml), then dried with nitrogen to form a film (45°C, rotary evaporator), and hydrated with deuterium-free water (45°C, 1 h, 200 rpm), with a deuterium content of ≤50 ppm; the liposome suspension is then sonicated with a probe (100 W, 20 min, ice bath), filtered through a 0.45 μm filter membrane, and the liposome suspension has a particle size of 80–200 nm and a PDI ≤0.3.

[0026] S4: Disperse the encapsulated complex from step S2 in the liposome suspension from step S3, and obtain a low-deuterium water-based anthocyanin complex stable system after lyophilization or sterile filtration.

[0027] In step S4, with a mass ratio of liposome suspension to encapsulation complex of 10:1, 2% glycerol (lyophilization protectant) is added when the anthocyanin-CD encapsulation complex is dispersed in the liposome solution, followed by vacuum lyophilization (-50℃, 0.1mbar, 24h) or aseptic filtration followed by nitrogen filling (headspace oxygen ≤1%).

[0028] Example 2: A method for preparing a low-deuterium water-based anthocyanin complex stabilization system The method for preparing a low-deuterium water-based anthocyanin complex stabilization system includes the following steps: S1: Obtain an anthocyanin-containing extract.

[0029] Take black goji berries, blueberries, and mulberries, wash them, freeze-dry them (-50℃, 24h), and then pulverize them through a 40-mesh sieve.

[0030] Ultrasonic extraction with low-deuterium water: Add low-deuterium water with a deuterium content ≤80ppm at a material-to-liquid ratio of 1:15 (w / v), and ultrasonically extract for 30 min at 40kHz and 200W, with an extraction temperature of 45-50℃ (temperature controlled by ice bath); centrifuge the extract at 8000rpm for 15 min and collect the supernatant.

[0031] S2: The anthocyanin extract from step S1 is encapsulated with β-cyclodextrin in deuterated water to obtain an anthocyanin-β-cyclodextrin encapsulated complex.

[0032] In situ encapsulation of β-cyclodextrin: β-cyclodextrin was directly added to the supernatant obtained in step S1 (the amount of supernatant added was determined by measuring the anthocyanin content in the supernatant, with anthocyanin:β-cyclodextrin = 1:2 molar ratio), and the mixture was stirred at 46°C for 2.5 h (200 rpm) for encapsulation. The mixture was then freeze-dried (-50°C, 0.1 mbar, 24 h) to obtain the anthocyanin-β-cyclodextrin encapsulated complex, with an encapsulation rate ≥85% (determined by UV-Vis method).

[0033] S3: The phospholipids were dissolved in anhydrous ethanol, dried under nitrogen to form a film, hydrated with deuterium-free water, and homogenized under high pressure to obtain a liposome suspension.

[0034] In step S3, the phospholipid is hydrogenated soybean lecithin, which is dissolved in anhydrous ethanol (50 mg / ml), then dried with nitrogen to form a film (45°C, rotary evaporator), and hydrated with deuterium-free water (45°C, 1 h, 200 rpm), with the deuterium content of the deuterium-free water ≤50 ppm; it is then homogenized under high pressure (100 MPa, 5 cycles) and filtered through a 0.45 μm filter membrane. The liposome suspension has a particle size of 80–200 nm and a PDI ≤0.3.

[0035] S4: Disperse the encapsulated complex from step S2 in the liposome suspension from step S3, and obtain a low-deuterium water-based anthocyanin complex stable system after lyophilization or sterile filtration.

[0036] In step S4, with a mass ratio of 9:1 between the liposome suspension and the encapsulation complex, 2% glycerol (lyophilization protectant) is added when the anthocyanin-CD encapsulation complex is dispersed in the liposome solution. Then, the mixture is lyophilized under vacuum (-50℃, 0.1mbar, 24h) or sterile filtered and filled with nitrogen (headspace oxygen ≤1%).

[0037] Example 3: A method for preparing a low-deuterium water-based anthocyanin complex stabilization system The method for preparing a low-deuterium water-based anthocyanin complex stabilization system includes the following steps: S1: Obtain an anthocyanin-containing extract.

[0038] Purple sweet potatoes were crushed and anthocyanins were extracted using acidic ethanol (0.1% HCl, pH 3.0, 60% ethanol) at 45℃ for 1 hour. The extracted anthocyanins were then purified by adsorption onto AB-8 macroporous resin and elution with 70% ethanol. The collected eluent was freeze-dried (-50℃, 0.1 mbar) to obtain anthocyanin freeze-dried powder with an anthocyanin content ≥80% (HPLC method, cyanidin-3-O-glucoside standard).

[0039] S2: The anthocyanin extract from step S1 is encapsulated with β-cyclodextrin in deuterated water to obtain an anthocyanin-β-cyclodextrin encapsulated complex.

[0040] In step S2, β-CD (β-cyclodextrin) is first dissolved in deuterated water (5% w / v, dissolved at 60℃, deuterium content of deuterated water ≤50ppm), and then the anthocyanin lyophilized powder prepared in step 1 (anthocyanin:β-CD = 1:3 molar ratio) is added, and the mixture is stirred and embedded (47℃ / 2.5h / 220rpm), and then freeze-dried to obtain the embedded complex (-50℃, 0.1mbar, 24h), with an embedding rate ≥85% (determined by UV-Vis method).

[0041] S3: The phospholipids are dissolved in anhydrous ethanol and dried with nitrogen to form a film. The film is then hydrated with deuterium-free water and homogenized by ultrasound or high pressure to obtain a liposome suspension.

[0042] In step S3, the phospholipid is soybean lecithin, which is dissolved in anhydrous ethanol (50 mg / ml), then dried with nitrogen to form a film (45°C, rotary evaporator), and hydrated with deuterium-free water (45°C, 1 h, 200 rpm), with the deuterium content of the deuterium-free water ≤ 50 ppm; after high-pressure homogenization (95 MPa, 6 cycles), and filtration through a 0.45 μm filter membrane, the liposome suspension has a liposome particle size of 80–200 nm and a PDI ≤ 0.3.

[0043] S4: Disperse the encapsulated complex from step S2 in the liposome suspension from step S3, and obtain a low-deuterium water-based anthocyanin complex stable system after lyophilization or sterile filtration.

[0044] In step S4, with a mass ratio of liposome suspension to encapsulation complex of 11:1, 2% glycerol (lyophilization protectant) is added when the anthocyanin-CD encapsulation complex is dispersed in the liposome solution, followed by vacuum lyophilization (-50℃, 0.1mbar, 24h) or aseptic filtration followed by nitrogen filling (headspace oxygen ≤1%).

[0045] In each embodiment of the method for preparing the low-deuterium water-based anthocyanin complex stabilization system described above, the following normative documents must be complied with: GB 2760 National Food Safety Standard for the Use of Food Additives; GB 2762 National Food Safety Standard for Limits of Contaminants in Food; GB 4789.2 National Food Safety Standard for Microbiological Examination of Food - Determination of Total Colony Count; GB 4789.3 National Food Safety Standard for Microbiological Examination of Food - Coliform Count; GB 4789.4 National Food Safety Standard for Microbiological Examination of Food - Salmonella Examination; GB 4789.10 National Food Safety Standard for Microbiological Examination of Food - Staphylococcus aureus Examination; GB 4789.15 National Food Safety Standard for Microbiological Examination of Food - Mold and Yeast Count; GB 5009.3 National Food Safety Standard for Determination of Moisture in Food; GB 5009.12 National Food Safety Standard for Determination of Lead in Food; GB 5749 Hygienic Standard for Drinking Water; GB 7718 National Food Safety Standard for General Rules for Labeling of Prepackaged Foods; GB 14881 National Food Safety Standards: General Hygiene Specifications for Food Production; GB 28050 National Food Safety Standards: General Rules for Nutrition Labelling of Prepackaged Foods; NY / T 2640 Determination of Anthocyanins in Plant-Based Foods by High Performance Liquid Chromatography; JJF 1070 Rules for Metrological Inspection of Net Content of Quantitatively Packaged Commodities.

[0046] Example 4: A low-deuterium water-based anthocyanin beverage Formula (per 1000L): 500-2000g of the low-deuterium water-based anthocyanin complex stabilizing system from Example 1, 200-500g of Vitamin C, an appropriate amount of citric acid-sodium citrate buffer system (to adjust pH 3.8-4.2), 20-50kg of erythritol, 50-200g of steviol glycosides, 100-500g of edible flavoring, and low-deuterium water (deuterium content ≤50ppm) to make up to 1000L. Optional additions include 100-500g of Polygonatum sibiricum polysaccharide extract or 5-15kg of fructooligosaccharides.

[0047] Preparation method: (1) Preparation of low-deuterium water: Natural water source is heated to 60-80℃ and evaporated under a vacuum of -0.08MPa using vacuum distillation. The condensate is collected and the deuterium content is tested to be ≤50ppm. Alternatively, food-grade low-deuterium water can be purchased directly.

[0048] (2) Compound flavoring: Dissolve the low-deuterium water-based anthocyanin complex stabilization system in low-deuterium water, and add vitamin C, citric acid-sodium citrate buffer, erythritol, steviol glycosides and flavoring in sequence, and stir evenly.

[0049] (3) Filling and sterilization: After being sterile filtered through 0.22μm, nitrogen filling (headspace oxygen ≤1%) is performed and the container is sealed to prevent light exposure.

[0050] The beverage retains 76.5% of anthocyanins at room temperature for 6 months (less than 50% in the control group without encapsulation in ordinary water), and 71.2% at 40℃ / 75%RH for 30 days (less than 35% in the control group without encapsulation in ordinary water). It has a shelf life of 12 months at room temperature.

[0051] The requirements for raw materials and auxiliary materials in the preparation of this low-deuterium water-based anthocyanin beverage are as follows: Vitamin C must comply with GB 2760; production water must comply with GB 5749 and be treated by RO reverse osmosis before use, with a conductivity ≤10μS / cm.

[0052] Sensory requirements for low-deuterium water-based anthocyanin beverages: uniform color, no layering, no sediment; taste and aroma with the fragrance of black goji berries, pleasantly sweet and sour, no off-flavors or bitterness; and a uniform liquid texture, free of visible impurities and suspended matter.

[0053] Physicochemical indicators of low-deuterium water-based anthocyanin beverage: soluble solids: ≤5°Brix; pH value: 3.8-4.0; anthocyanin content: ≥20mg / 100ml; lead (as Pb): ≤0.3mg / kg; arsenic (as As): ≤0.1mg / kg; The microbiological indicators are as follows: total bacterial count ≤100 CFU / ml; coliform bacteria not detected; Salmonella not detected; Staphylococcus aureus not detected; molds and yeasts ≤10 CFU / ml.

[0054] Food additives: The scope of use and maximum usage of all food additives shall comply with the provisions of GB 2760, and they shall not be used beyond the scope or limit.

[0055] Hygiene requirements for production and processing: The production process complies with GB 14881. The mixing and filling processes are carried out in a cleanroom with a cleanliness level of not less than 100,000. All equipment, pipes and containers that come into contact with the products meet the requirements of food-grade materials and are cleaned and disinfected by CIP before use. Anthocyanin products are handled in the dark throughout the process, and brown storage tanks and aluminum foil-wrapped pipes are used to prevent light from causing anthocyanin degradation.

[0056] Test methods: Soluble solids were determined according to the method specified in GB / T 12143; pH value was determined according to the method specified in GB / T 12456; anthocyanin content was determined according to the high performance liquid chromatography method specified in NY / T 2640, with sample pretreatment using 0.1% hydrochloric acid methanol solution for extraction in the dark, using a C18 reversed-phase column, and a gradient elution of 0.1% formic acid water and acetonitrile, with a detection wavelength of 520 nm; microbiological indicators were determined according to the methods specified in the GB 4789 series of standards; heavy metal indicators were determined according to the methods specified in GB 5009.12.

[0057] Inspection Rules: Batching Rules: Products from the same shift, production line, variety, and specification constitute a batch; Sampling Rules: Three bottles of finished product are randomly selected from each batch: one bottle for sensory and physicochemical testing, one bottle for microbiological testing, and one bottle is retained as a sample. Judgment Rules: If the sensory, physicochemical, and microbiological indicators all meet the requirements of this standard, the batch of products is deemed qualified; if any indicator fails to meet the standard, the sampling is doubled for re-inspection. If the re-inspection still fails to meet the standard, the batch of products is deemed unqualified.

[0058] Marking, packaging, transportation, and storage rules: The labeling must comply with GB 7718 and GB 28050. Labels must include warnings such as "no added sugar," "rich in anthocyanins," "recommended daily intake of 1 bottle (300ml)," "use with caution if you have a weak spleen and stomach," and "this product cannot replace medication." Labels must not claim medical effects such as "improves eyesight," "strengthens the brain," or "strengthens the spleen." Packaging must be in dark-colored PET bottles or aluminum cans with a well-sealed cap. Outer packaging must comply with GB / T191. Transportation must be clean, dry, and odorless, avoiding direct sunlight, rain, and high temperatures. It must not be transported with toxic, harmful, or odorous items. Store in a cool, dry, and well-ventilated place, avoiding direct sunlight. Refrigerated storage (2-8℃) has a shelf life of 6 months, while room temperature storage (≤25℃) has a shelf life of 3 months.

[0059] Supplementary Implementation Notes: 1. When coordinating with the contract manufacturer, they must provide the above procedures and third-party testing reports for raw materials, confirming that the manufacturer has experience in producing anthocyanin products and possesses aseptic cold filling / UHT instantaneous sterilization capabilities. 2. During small-batch trial production, first prepare a 500ml sample according to the formula and conduct a 7-day stability test (room temperature / refrigeration), testing anthocyanin retention rate, pH changes, and microbial indicators. Large-scale production can only proceed after confirming no precipitation, no discoloration, and stable taste. 3. If formula adjustments are necessary, the proportion of the low-deuterium water-based anthocyanin complex stabilizing system can be fine-tuned by adding 20%-25% to the original ratio. After each adjustment, stability testing and sensory blind testing must be conducted again to ensure a balance between efficacy and taste. 4. All production and inspection records must be completely preserved for at least 6 months after the product's expiration date for quality traceability and compliance review.

[0060] Example 5: A low-deuterium water-anthocyanin-Polygonatum polysaccharide synergistic antioxidant composition Formula (mass percentage): 5% of the low-deuterium water-based anthocyanin complex stabilizing system from Example 1, 5% of Polygonatum sibiricum polysaccharide extract (polysaccharide ≥ 50%), the mass ratio of the low-deuterium water-based anthocyanin complex stabilizing system to the Polygonatum sibiricum polysaccharide extract is 1:1, vitamin C 0.02%, zinc glycine 0.001%, and low-deuterium water to make up to 100%.

[0061] Preparation method: Dissolve each component in deuterium-free water according to the above formula, stir evenly, filter aseptically and fill into powder, or freeze-dry to make powder, or fill into capsules.

[0062] Mechanism of action: As a health food, this composition has three antioxidant pathways: low-deuterium water activates endogenous antioxidant enzymes such as SOD and CAT in cells; anthocyanins directly scavenge free radicals; and Polygonatum polysaccharides synergistically enhance the activity of SOD and GSH-Px enzymes.

[0063] Example 6: A low-deuterium water-based health tea with synergistic regulation of high blood pressure, high cholesterol, and high blood sugar. Formula (approximately per 500mL of finished product): 450mL of low-deuterium water (≤50ppm), 5g of the low-deuterium water-based anthocyanin complex stabilizing system from Example 2, 1.2g of freeze-dried black goji berry powder (anthocyanins ≥25%), 10mL of concentrated ultrasonic extract of Polygonatum sibiricum (polysaccharides ≥50%, cycloartenol ≥92%), 15mL of enzymatic hydrolysis fermentation broth of mulberry (proanthocyanins ≥15%), 0.8g of supercritical CO2 extract of hawthorn (flavonoids ≥30%), and 5mL of water-extracted alcohol precipitate of cassia seed (anthraquinones ≤0.3%).

[0064] Preparation method: (1) Stepwise targeted extraction: Polygonatum was extracted and concentrated under the conditions of 50kHz ultrasonic assistance, 60℃ and material-liquid ratio of 1:15; Mulberry was fermented by double enzymatic hydrolysis of pectinase and cellulase (pH4.2, 42℃); Hawthorn was extracted by supercritical CO2 at 35MPa and 45℃; Cassia seed was extracted with water at 80℃, phenol was removed by AB-8 macroporous resin and alcohol precipitation; All components were tested and qualified before use.

[0065] (2) Low temperature compounding: Under the protection of argon or nitrogen, the entire operation temperature is ≤4℃. The extracts are added to the low deuterium water in the following order: Polygonatum extract → Mulberry extract → Hawthorn extract → Cassia seed extract → Black goji berry powder → Low deuterium water-based anthocyanin complex stabilization system. Each step is stirred for 5 minutes (200 rpm).

[0066] Low-deuterium water-based health tea that synergistically regulates blood sugar and blood pressure can be used to regulate these conditions.

[0067] Example 7: A low-deuterium water-based stomach-nourishing rice paste Formula (by weight): 40 parts millet, 20 parts black rice, 15 parts monkey head mushroom, 15 parts yam, 10 parts polygonatum, 5 parts black goji berries, 5 parts mulberry, 10 parts red dates, 10 parts oats, 15 parts milk powder, and 10 parts of the low-deuterium water-based anthocyanin complex stabilizing system from Example 2.

[0068] Preparation of powder-based reconstituted formulations: (1) Ultra-fine cell wall breaking: Hericium erinaceus dried slices and yam were freeze-dried (-50℃, 24h) and then air-jet pulverized to D90≤10μm (the bitterness of yam was reduced by more than 60%); millet and black rice were pulverized and passed through a 200-mesh sieve.

[0069] (2) Mixing and packaging: Each ingredient is mixed in a V-type mixer (20 rpm, 15 min), and the low deuterium water-based anthocyanin complex stabilization system is added last and mixed for no more than 5 min. The mixture is then packaged in nitrogen-filled aluminum foil vacuum bags (30 g / bag).

[0070] Preparation of bottled ready-to-drink type: (1) Ultra-fine cell wall breaking: Hericium erinaceus dried slices and yam were freeze-dried (-50℃, 24h) and then air-jet pulverized to D90≤10μm (the bitterness of yam was reduced by more than 60%); millet and black rice were pulverized and passed through a 200-mesh sieve.

[0071] (2) Mixing: Each ingredient is mixed in a V-type mixer (20 rpm, 15 min). The low-deuterium water-based anthocyanin composite stabilization system is added last and mixed for no more than 5 min to obtain powder.

[0072] (3) Mixing and homogenizing: Mix powder and low-deuterium water at a ratio of 1:10 (by mass), add β-cyclodextrin (0.5% w / v), and homogenize under high shear (20 MPa, 3 times, material temperature ≤45℃).

[0073] (4) UHT sterilization: instant sterilization at 135℃ / 4s, followed by immediate cooling to below 25℃.

[0074] (5) Aseptic filling: nitrogen filling (headspace oxygen ≤2%), NFC four-dimensional freshness monitoring label on bottle body, and product digital identity written before leaving the factory.

[0075] This low-deuterium water-based stomach-nourishing rice cereal has a protective effect on the gastric mucosa.

[0076] Example 8: A low-deuterium water-based anthocyanin-rich fresh beer Raw materials: Low-deuterium water with a deuterium content ≤50ppm was used for saccharification and all process water; barley malt, hops, and yeast were conventional beer raw materials; anthocyanins were derived from the low-deuterium water-based anthocyanin complex stabilization system in Example 3. The finished product had an alcohol content of 4–6% vol and a pH of 4.0–4.5.

[0077] Preparation methods: (1) Preparation of low-deuterium water for brewing: vacuum distillation or purchase food-grade low-deuterium water (≤50ppm). (2) Saccharification of wort: after crushing barley malt, saccharify at 65℃ (material-to-water ratio 1:4), filter, boil and add hops, and cool to obtain wort. (3) Primary fermentation and post-fermentation: inoculate the wort with yeast, primary ferment at 18-22℃ for 5-7 days, and post-fermentation at 0-4℃ for 7-14 days. (4) Post-modification and addition of anthocyanins: after filtering, disperse the low-deuterium water-based anthocyanin complex stabilization system in the beer to avoid contact with SO2 produced during fermentation and bleaching, and stir gently at 200rpm for 15min. (5) Nitrogen-filled / micro-frozen preservation and bottling: nitrogen-filled bottling (headspace oxygen ≤1%), micro-frozen preservation (0±1℃ cold chain) can be selected, and sealed in the dark.

[0078] This fresh beer retains ≥70% of anthocyanins after 60 days of refrigeration (4-8℃) (compared to ≤40% for ordinary anthocyanin beer and severe fading), resulting in a purplish-red color without browning; the low deuterium content reduces fusel alcohol content, significantly reducing the throat feel.

[0079] Example 9: A low-deuterium water-based anthocyanin health-preserving baijiu Base liquor: Select light-aroma or strong-aroma medium-low alcohol content baijiu (20-38% vol).

[0080] Preparation method: (1) Take the low-deuterium water-based anthocyanin composite stabilizing system from Example 3 and use it with -Zn 2+ Complexation: Using deuterium-rich water as a medium, a zinc gluconate and a deuterium-rich water-based anthocyanin complex stabilizing system were added to the system. The amounts of the deuterium-rich water-based anthocyanin complex stabilizing system and zinc gluconate added were based on the anthocyanin:Zn ratio in the system. 2+ =1∶0.5 (molar ratio), which allows the zinc gluconate and low-deuterium water-based anthocyanin complex to form a stable Zn complex system. 2+ - Anthocyanin complexes are obtained to form an anthocyanin complex system, which enhances the color stability and antioxidant properties of the wine. (2) Blending: The base wine is diluted with deuterium water to the target alcohol content, and the above anthocyanin complex system is added and stirred gently. (3) Aging and bottling: Aging is carried out at room temperature in the dark for 7 to 15 days, and then bottled after being sterile filtered through 0.22 μm.

[0081] The prepared liquor retains ≥75% anthocyanins after 6 months (ordinary anthocyanin liquor ≤45%, which is prone to precipitation and discoloration), and the liquor is purplish-red and clear without turbidity or sediment; the low deuterium water dilution reduces fusel alcohols, and it does not cause headaches after drinking.

[0082] Comparative Experiment 1: Stability Comparison of Different Protection Systems To verify the triple synergistic effect of the low-deuterium water-based anthocyanin composite stabilization system of this invention, the following control group was set up. An accelerated test was conducted at 60℃ for 30 days, and the anthocyanin retention rate was measured. The results are shown in Table 1: Table 1 Comparison of anthocyanin retention rates in different protection systems (accelerated test at 60℃ / 30 days) Control group A Unprotected — — — 12.3 Control group B Only β-cyclodextrin encapsulation √ — — 62.3 Control group C Liposome encapsulation only — √ — 58.7 Control group D β-Cyclodextrin + Liposomes (ordinary water) √ √ — 71.8 experimental group β-Cyclodextrin + Liposomes + Low-Deuterium Water (This Invention) √ √ √ 85.1 As shown in Table 1, the retention rates of anthocyanins were as follows: Control group A (unprotected): 12.3% when anthocyanins were directly dissolved in ordinary water; Control group B (β-cyclodextrin encapsulation only): 62.3%; Control group C (liposome encapsulation only): 58.7%; Control group D (β-cyclodextrin + liposomes, ordinary water): 71.8%; and Experimental group (β-cyclodextrin + liposomes + low-deuterium water, the low-deuterium water-based anthocyanin composite stabilization system prepared in this embodiment of the invention): 85.1%.

[0083] The results show that the retention rate of the triple protection system of this invention is significantly higher than that of any single protection or double protection combination, and the synergistic effect multiple is 85.1% / 62.3% = 1.37 times, proving that there is a significant synergistic effect between deuterium water and β-cyclodextrin and liposomes.

[0084] Comparative Experiment 2: Long-term stability test The low-deuterium water-based anthocyanin composite stabilization system prepared in the embodiments of the present invention was investigated under different conditions. The results are shown in Table 2. As can be seen from Table 2, the retention rate was 86.7% at 40℃ for 90 days; 92.3% at 25℃ for 180 days; 85.1% at 60℃ for 30 days; and 79.8% after 5 cycles in environments ranging from pH 3 to pH 7. All indicators are significantly better than the existing technology.

[0085] Table 2. Long-term stability test results of the composite stabilization system of the present invention 1 40℃ accelerated test 90 days 86.7 2 25℃ room temperature test 180 days 92.3 3 60℃ accelerated test 30 days 85.1 4 pH 3 → pH 7 cycle (5 times) — 79.8 Note: All of the above indicators are significantly better than the existing technology level (the existing technology has a retention rate of 35% to 50% at 40℃ / 90 days and a retention rate of less than 60% at 25℃ / 180 days).

[0086] Comparative Example 3: Bioavailability Test The transmembrane uptake rate of anthocyanins was evaluated using the Caco-2 cell model. The results are shown in Table 3. As can be seen from Table 3, the cell uptake rate of the ordinary water control group was 70%, while the uptake rate of the low-deuterium water-based triple stabilization system of this invention was 82%, an increase of 12 percentage points. This indicates that the low-deuterium water carrier and liposome encapsulation synergistically promoted the cell uptake of anthocyanins.

[0087] Table 3 Comparison of anthocyanin transmembrane uptake rates in Caco-2 cell models control group Ordinary water (without triple protection) 70 — experimental group Low-deuterium water-based triple stabilization system (this invention) 82 +12 percentage points It should be noted that, in this document, terms such as “comprising,” “including,” or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0088] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a low-deuterium water-based anthocyanin complex stable system, characterized in that, Includes the following steps: S1: Obtain an anthocyanin-containing extract; S2: The anthocyanin extract from step S1 was encapsulated with β-cyclodextrin in deuterated water to obtain an anthocyanin-β-cyclodextrin encapsulated complex. S3: Dissolve phospholipids in anhydrous ethanol, dry them with nitrogen to form a film, hydrate them with deuterium-free water, and homogenize them by ultrasound or high pressure to obtain a liposome suspension. S4: Disperse the encapsulated complex from step S2 in the liposome suspension from step S3, and obtain a low-deuterium water-based anthocyanin complex stable system after lyophilization or sterile filtration.

2. The method for preparing a low-deuterium water-based anthocyanin composite stable system according to claim 1, characterized in that, In step S1, the method for obtaining the anthocyanin-containing extract is as follows: Using at least one of black goji berries, purple sweet potatoes, blueberries, and mulberries as raw materials, anthocyanins are extracted and purified by acidic ethanol to obtain freeze-dried powder with an anthocyanin content ≥80%. or, Using at least one of black goji berries, purple sweet potatoes, blueberries, and mulberries as raw materials, and using low-deuterium water with a deuterium content ≤80ppm instead of acidic ethanol as the extraction solvent, the anthocyanin raw materials are subjected to ultrasonic-assisted extraction to obtain anthocyanin extract, which is then used in step S2 to directly add β-cyclodextrin to the anthocyanin extract for in-situ encapsulation.

3. The method for preparing a low-deuterium water-based anthocyanin composite stable system according to claim 1, characterized in that: In step S2, the molar ratio of anthocyanins to β-cyclodextrin in the obtained anthocyanin-β-cyclodextrin encapsulation complex is 1:2 to 1:3, and the encapsulation rate is ≥85%; in step S3, the liposomes in the liposome suspension have a particle size of 80 to 200 nm and a polydispersity index ≤0.3; the deuterium content of the low-deuterium water in steps S2 and S3 is ≤50 ppm.

4. The method for preparing a low-deuterium water-based anthocyanin complex stabilization system according to claim 1, characterized in that: In step S2, the embedding temperature is 45±2℃, the embedding time is 1.5~2.5h, and the embedding reaction is carried out by stirring at a speed of 180~220rpm. In step S3, the phospholipid is soybean lecithin or hydrogenated soybean lecithin, and the zeta potential of the liposome is -30 to -50 mV; the high-pressure homogenization conditions in step S3 are 95 to 105 MPa and 4 to 6 cycles. In step S4, the embedding complex and liposome suspension are added in a mass ratio of 9:1 to 11:1 between the liposome suspension and the embedding complex.

5. The low-deuterium water-based anthocyanin complex stabilizing system prepared by the method for preparing a low-deuterium water-based anthocyanin complex stabilizing system according to any one of claims 1-4.

6. A low-deuterium water-based anthocyanin beverage, characterized in that: The system includes the low-deuterium water-based anthocyanin composite stabilization system as described in claim 5, using low-deuterium water with a deuterium content ≤50ppm as the matrix, the addition amount of the low-deuterium water-based anthocyanin composite stabilization system is 500~2000g / 1000L, the pH value is 3.8~4.2, nitrogen filling is used and the headspace oxygen is ≤1%.

7. A low-deuterium water-anthocyanin-Polygonatum polysaccharide synergistic antioxidant composition, characterized in that: The invention comprises the low-deuterium water-based anthocyanin complex stabilizing system as described in claim 5 and Polygonatum sibiricum polysaccharide extract, wherein the polysaccharide content in the Polygonatum sibiricum polysaccharide extract is ≥50%, and the mass ratio of the low-deuterium water-based anthocyanin complex stabilizing system to the Polygonatum sibiricum polysaccharide extract is 1:1 to 1:

5.

8. A low-deuterium water-based health-preserving tea with synergistic regulation of high blood pressure, high cholesterol, and high blood sugar, characterized in that... The product comprises the low-deuterium water-based anthocyanin complex stabilizing system as described in claim 5, as well as black goji berry freeze-dried powder, polygonatum ultrasonic extraction concentrate, mulberry enzymatic fermentation broth, hawthorn supercritical CO2 extract and cassia seed water-extracted alcohol precipitate, and each component is compounded in a gradient manner under ≤4℃ and inert gas protection.

9. A low-deuterium water-based stomach-nourishing rice paste, characterized in that, The product comprises the low-deuterium water-based anthocyanin complex stabilizing system as described in claim 5, and a complex of millet, black rice, Hericium erinaceus, yam, Polygonatum sibiricum, black goji berries, mulberries, red dates, oats, and milk powder or yogurt, wherein the Hericium erinaceus is pulverized by airflow to a D90≤10μm; the rice paste is a powder-based reconstituted type or a bottled ready-to-drink type.

10. A low-deuterium water-based anthocyanin-rich fresh beer, characterized in that: The beer is brewed using low-deuterium water-based anthocyanin complex stabilizing system as described in claim 5, with a deuterium content of ≤50ppm as the brewing water. After filtration, the low-deuterium water-based anthocyanin complex stabilizing system is added to the beer as a modifier. The alcohol content is 4-6% vol, and the beer is bottled using nitrogen filling or micro-freezing preservation.

11. A low-deuterium water-based anthocyanin health-preserving baijiu, characterized in that: The system comprises the low-deuterium water-based anthocyanin complex stabilizing system as described in claim 5, using 20-38% vol medium-low alcohol baijiu as the base liquor, which is diluted and blended with low-deuterium water, and then processed through anthocyanin-Zn... 2+ The complex and the low-deuterium water-based anthocyanin complex stabilization system were modified and then bottled and stored.