Anti-aging oral liquid and preparation method thereof

Abstract

The invention provides anti-aging oral liquid and a preparation method thereof, and belongs to the technical field of oral liquid. Ginseng, turmeric, astragalus membranaceus and ginger are subjected to bidirectional fermentation through a ganoderma lucidum seed solution, a product is subjected to alcohol precipitation to obtain traditional Chinese medicine polysaccharide, and active components are obtained through freeze drying; the preparation method comprises the following steps: extracting cod skin to obtain a collagen peptide-copper compound, mixing the collagen peptide-copper compound with egg yolk protein for enzymolysis to obtain an egg yolk protein-collagen peptide-copper composition, adding the egg yolk protein-collagen peptide-copper composition into water, embedding the egg yolk protein-collagen peptide-copper composition with active components to obtain an active component embedding solution, adding traditional Chinese medicine polysaccharide, resveratrol, taurine and lycopene, homogenizing and filtering to obtain the anti-aging oral liquid. The anti-aging oral liquid prepared by the invention is simple in preparation method and wide in raw material source, and the prepared oral liquid has excellent anti-oxidation, anti-aging and anti-cancer effects, is relatively low in cost, is easy to realize industrial production, and has a wide application prospect.

Description

Technical Field

[0001] This invention relates to the field of oral liquid technology, specifically to an anti-aging oral liquid and its preparation method. Background Technology

[0002] Human aging is partly caused by lipid peroxidation triggered by free radicals, producing metabolic byproducts such as lipid peroxides (LPO). These byproducts form brown substances that accumulate in various organs, ultimately leading to damage to cellular physiological functions and potentially causing decreased immunity and memory decline. These free radicals include oxygen free radicals, hydrogen free radicals, and hydrogen peroxide, which are widely present in body tissues. They attack unsaturated fatty acids in cell membranes and mitochondrial membranes, causing enhanced lipid peroxidation. The products of lipid peroxidation can then decompose into more free radicals, triggering a chain reaction that causes widespread damage to brain cells, muscle cells, and other organs. Under normal circumstances, free radicals in the human body are in a dynamic balance between continuous production and elimination. Excessive production or slow elimination of free radicals can cause various types of damage at the molecular, cellular, and organ levels by attacking macromolecules and various cells, accelerating the aging process and inducing various diseases. While the human body inevitably produces free radicals, it also naturally produces antioxidants to counteract the oxidative damage caused by free radicals to human cells. Studies have shown that the body's antioxidant system and immune system are both important systems for any health condition, and the relationship between an organism's antioxidant capacity and health is becoming increasingly important.

[0003] With the development of science and technology, various peroxides may be generated in production and daily life; radiation from mobile phones, televisions, computers, etc., can also produce free radicals. In addition, modern people face high pressure, fast-paced lifestyles, irregular eating habits, and sub-health conditions caused by insufficient sleep. Those who engage in strenuous labor or high-intensity exercise, work in oxygen-deficient environments, have low immunity, or have high blood lipids are also prone to oxygen deficiency. All of these factors can lead to an imbalance in the body's antioxidant system, thereby damaging the integrity and stability of human cells. This damage to the human body is medically termed "oxidative damage." How to enhance immunity and improve the body's antioxidant capacity has become a concern for the entire society.

[0004] Therefore, in food production, chemically synthesized antioxidants are often used to eliminate the effects of free radicals. While effective, these antioxidants carry inherent safety risks. Consequently, people tend to prefer natural, reliable, and safe antioxidant products. Existing technologies extensively utilize natural substances as antioxidant components in oral liquid formulations, such as oral compositions using black garlic and cordyceps. However, these generally suffer from drawbacks including complex preparation processes, less effective results, and high costs. Summary of the Invention

[0005] The purpose of this invention is to propose an anti-aging oral liquid and its preparation method. The preparation method is simple, the raw materials are widely available, the oral liquid has excellent antioxidant, anti-aging and anti-cancer effects, and the cost is low. It is easy to realize industrial production and has broad application prospects.

[0006] The technical solution of this invention is implemented as follows:

[0007] This invention provides a method for preparing an anti-aging oral liquid. Ginseng, turmeric, astragalus, and ginger are fermented in a bidirectional manner using Ganoderma lucidum seed liquid. The product is precipitated with alcohol to obtain a traditional Chinese medicine polysaccharide, which is then freeze-dried to obtain the active component. Cod skin is extracted to obtain a collagen peptide-copper complex, which is mixed with egg yolk protein and enzymatically hydrolyzed to obtain an egg yolk protein-collagen peptide-copper composition. This composition is added to water and encapsulated with the active component to obtain an active ingredient encapsulation solution. Traditional Chinese medicine polysaccharides, resveratrol, taurine, and lycopene are added, and the mixture is homogenized and filtered to obtain the anti-aging oral liquid.

[0008] As a further improvement to the present invention, the following steps are included:

[0009] S1. Preparation of collagen peptide-copper complex: Cod skin was scaled, washed, minced, added to water, enzymatically hydrolyzed with protease, copper salt was added, the reaction was stirred, filtered, the filtrate was dialyzed, and freeze-dried to obtain collagen peptide-copper complex.

[0010] S2. Two-way fermentation of traditional Chinese medicine: Ginseng, turmeric, astragalus and ginger are washed, crushed and mixed to obtain traditional Chinese medicine powder. Add water, glucose, peptone and sodium chloride, stir and mix evenly, sterilize, inoculate with Ganoderma lucidum seed liquid, ferment and culture, filter, and the filtrate is the fermentation product.

[0011] S3. Preparation of polysaccharides from traditional Chinese medicine: Ethanol is added to the fermentation product obtained in step S2 for precipitation, filtered, and the solid is dried to obtain polysaccharides from traditional Chinese medicine. The filtrate is freeze-dried to obtain the active component.

[0012] S4. Preparation of egg yolk protein-collagen peptide-copper composition: Add egg yolk protein to water, add the collagen peptide-copper complex obtained in step S1, adjust the pH of the solution to alkaline, add trypsin, heat to hydrolyze, adjust the pH of the solution to neutral, inactivate the enzyme, filter, freeze-dry the filtrate to obtain egg yolk protein-collagen peptide-copper composition.

[0013] S5. Preparation of active ingredient encapsulation solution: Add the egg yolk protein-collagen peptide-copper composition obtained in step S4 to water, stir and mix, hydrate overnight, adjust the pH of the solution to alkaline, add the active component obtained in step S3, stir and mix, adjust the pH of the solution to neutral, centrifuge, remove solids, and obtain active ingredient encapsulation solution.

[0014] S6. Preparation of additives: Resveratrol, taurine and lycopene are mixed evenly to prepare additives;

[0015] S7. Preparation of anti-aging oral liquid: The traditional Chinese medicine polysaccharide obtained in step S3 and the additive obtained in step S6 are added to the active ingredient encapsulation solution obtained in step S5, homogenized, filtered, and the anti-aging oral liquid is obtained.

[0016] As a further improvement of the present invention, the protease in step S1 is selected from at least one of trypsin, pepsin, papain, neutral protease, bromelain, alkaline protease, and fig protease. Preferably, it is a mixture of papain and bromelain in a mass ratio of 7-10:3-5. The enzymatic hydrolysis temperature is 40-50°C and the time is 2-4 hours. The mass ratio of cod skin, protease, and copper salt is 100:2-4:1-2. The copper salt is selected from at least one of copper chloride, copper sulfate, and copper nitrate. The dialysis is performed using a dialysis bag with a pore size of 3-5 kDa and the dialysis time is 3-5 hours.

[0017] As a further improvement of the present invention, the mass ratio of ginseng, turmeric, astragalus, ginger, glucose, peptone, and sodium chloride in step S2 is 1-2:7-10:3-5:3-5:7-12:5-7:2-3, the inoculation amount of the Ganoderma lucidum seed liquid is 8-10%, the mass fraction of the Ganoderma lucidum seed liquid is 8-10%, and the fermentation culture conditions are 28-32℃, 100-150r / min, and culture for 5-7 days.

[0018] As a further improvement of the present invention, in step S3, ethanol is added until the ethanol content of the system is 70-80 wt%, and the precipitation time is 3-5 h.

[0019] As a further improvement of the present invention, the mass ratio of egg yolk protein, collagen peptide-copper complex and trypsin in step S4 is 10-12:7-10:0.3-0.5, the pH value of the solution is adjusted to be alkaline in the specific range of 7.5-8.5, the temperature of the heating hydrolysis is 35-45℃, and the time is 1-3h.

[0020] As a further improvement of the present invention, the mass ratio of the egg yolk protein-collagen peptide-copper composition and the active component in step S5 is 15-22:3-5, the overnight hydration temperature is 3-5°C, and the pH of the solution is adjusted to be alkaline within the range of 10-12.

[0021] As a further improvement of the present invention, the mass ratio of resveratrol, taurine and lycopene in step S6 is 3-5:1-2:2-4; the mass ratio of traditional Chinese medicine polysaccharide, additive and active ingredient encapsulation solution in step S7 is 5-7:1-3:100; the homogenization speed is 10000-12000 r / min and the time is 5-10 min.

[0022] As a further improvement to the present invention, the specific steps include:

[0023] S1. Preparation of collagen peptide-copper complex: 100 parts by weight of cod skin were descaled, washed, minced, added to 500 parts by weight of water, 2-4 parts by weight of protease were added, and enzymatic hydrolysis was carried out at 40-50℃ for 2-4 hours. 1-2 parts by weight of copper salt were added, and the mixture was stirred at 35-45℃ for 0.5-1 hours. The mixture was filtered, and the filtrate was dialyzed through a dialysis bag with a pore size of 3-5 kDa for 3-5 hours. The mixture was then freeze-dried to obtain collagen peptide-copper complex.

[0024] The protease is a mixture of papain and bromelain in a mass ratio of 7-10:3-5;

[0025] S2. Two-way fermentation of traditional Chinese medicine: Wash and crush 1-2 parts by weight of ginseng, 7-10 parts by weight of turmeric, 3-5 parts by weight of astragalus, and 3-5 parts by weight of ginger, mix them to obtain traditional Chinese medicine powder, add 500 parts by weight of water, add 7-12 parts by weight of glucose, 5-7 parts by weight of peptone, and 2-3 parts by weight of sodium chloride, stir and mix evenly, sterilize, inoculate with Ganoderma lucidum seed liquid, the inoculation amount is 8-10%, the mass fraction of the Ganoderma lucidum seed liquid is 8-10%, 28-32℃, 100-150r / min, ferment and culture for 5-7 days, filter, the filtrate is the fermentation product;

[0026] S3. Preparation of polysaccharides from traditional Chinese medicine: Add ethanol to the fermentation product obtained in step S2 until the ethanol content of the system is 70-80 wt%, precipitate for 3-5 hours, filter, dry the solid to obtain polysaccharides from traditional Chinese medicine, freeze-dry the filtrate to obtain the active component;

[0027] S4. Preparation of egg yolk protein-collagen peptide-copper composition: 10-12 parts by weight of egg yolk protein were added to 200 parts by weight of water, 7-10 parts by weight of the collagen peptide-copper complex obtained in step S1 were added, the pH of the solution was adjusted to 7.5-8.5, 0.3-0.5 parts by weight of trypsin were added, the mixture was heated to 35-45℃, stirred and hydrolyzed for 1-3 hours, the pH of the solution was adjusted to neutral, the enzyme was inactivated, the mixture was filtered, and the filtrate was freeze-dried to obtain the egg yolk protein-collagen peptide-copper composition;

[0028] S5. Preparation of active ingredient encapsulation solution: Add 15-22 parts by weight of the egg yolk protein-collagen peptide-copper composition obtained in step S4 to 200 parts by weight of water, stir and mix, hydrate overnight at 3-5℃, adjust the pH of the solution to 10-12, add 3-5 parts by weight of the active component obtained in step S3, stir and mix, adjust the pH of the solution to 7, centrifuge, remove solids, and obtain active ingredient encapsulation solution;

[0029] S6. Preparation of additive: Mix 3-5 parts by weight of resveratrol, 1-2 parts by weight of taurine and 2-4 parts by weight of lycopene evenly to prepare the additive.

[0030] S7. Preparation of anti-aging oral liquid: Add 5-7 parts by weight of the Chinese herbal polysaccharide obtained in step S3 and 1-3 parts by weight of the additive obtained in step S6 to 100 parts by weight of the active ingredient encapsulation solution obtained in step S5, homogenize at 10000-12000 r / min for 5-10 min, filter, and obtain the anti-aging oral liquid.

[0031] This invention further protects an anti-aging oral liquid prepared by the above-described preparation method.

[0032] The present invention has the following beneficial effects:

[0033] This invention employs bidirectional fermentation to process traditional Chinese medicine raw materials, achieving effects such as detoxification, preservation of efficacy, enhanced medicinal properties, and increased extraction rate. The enzyme system produced after fermentation of Ganoderma lucidum seed liquid can degrade the cell walls of the medicinal materials, promoting the release of effective components. Simultaneously, the tissues and components of the fungus are altered by enzymes in the Ganoderma lucidum, generating new components. Furthermore, the medicinal material matrix provides the nutrients needed by the Ganoderma lucidum seeds, promoting the growth of the fungus and resulting in abundant Ganoderma lucidum polysaccharides and active components such as Ganoderma lucidum saponins. During the fermentation process, ginseng, under the action of enzymes, converts ordinary saponins into highly active saponins such as Rb1, Rg3, and F1. Ginsenoside F1 inhibits cell senescence by inhibiting p38MAPK-dependent NF-κB activity and can also inhibit the migration of glioblastoma. Ginsenoside Rb1 can reduce the phosphorylation level of mTOR protein in the brain, exhibiting anti-aging activity. Ginsenoside Rg3 can increase autophagy and NAD+. +This protects against oxidative stress-induced cellular senescence. However, many ginsenosides are poorly soluble in water. Curcumin, a large amount dissolved during turmeric fermentation, possesses various pharmacological effects such as antioxidant and immunomodulatory properties. However, due to its poor water solubility, curcumin is poorly absorbed and metabolized rapidly, resulting in low bioavailability in vivo. 6-Gingerol, a large amount dissolved from ginger, is a polyphenol complex with antioxidant, anti-inflammatory, and cardiovascular protective effects such as lowering blood lipids and blood pressure, thus exhibiting good anti-aging and anti-tumor effects. However, it also suffers from poor water solubility and consequently, poor bioavailability. Therefore, this invention embeds protein-based nanobundles around these active components. The protein-based nanoparticles are non-toxic, biodegradable, and have good biocompatibility. The resulting egg yolk protein-collagen peptide-copper composition self-assembles with the active components through non-covalent interactions to form nanoparticles, improving their water solubility and bioavailability, thereby significantly enhancing efficacy, reducing raw material usage, and lowering preparation costs.

[0034] Meanwhile, the collagen peptide-copper complex and egg yolk peptide prepared by this invention also have excellent antioxidant effects and exhibit synergistic effects. In particular, the collagen peptide-copper complex significantly improves the body's absorption and utilization efficiency of copper ions. Copper is an essential trace element in the human body and can participate in the preparation of copper-containing metallothioneins, superoxide dismutase, and other strong antioxidant enzymes, protecting cells from oxidative damage and thus greatly enhancing the anti-aging effect.

[0035] Traditional Chinese medicine polysaccharides include astragalus polysaccharide, ginseng polysaccharide, turmeric polysaccharide, and ginger polysaccharide. Astragalus polysaccharide possesses various pharmacological effects, such as increasing telomerase activity, antioxidant, anti-inflammatory, immunomodulatory, and anti-tumor effects, thus delaying aging by protecting cells and scavenging free radicals. Ginseng polysaccharide exhibits multiple pharmacological activities, including enhancing immunity, delaying aging, anti-fatigue, and anti-tumor effects, delaying aging by scavenging free radicals, improving immunity, and resisting lipid peroxidation. Turmeric polysaccharide can inhibit the production and release of inflammatory cytokines, reduce inflammatory responses, scavenge free radicals, reduce oxidative stress, protect cells from damage and aging, promote the production and activity of immune cells, and enhance the function of the immune system. Ginger polysaccharide can also enhance anti-aging effects through anti-inflammatory and antioxidant properties.

[0036] Among the additives, resveratrol, taurine, and lycopene are also natural products with good antioxidant effects. Resveratrol can regulate SIRT1 by activating the calorie restriction effect, thus delaying aging. Taurine has certain antioxidant properties, neutralizing free radicals, reducing oxidative stress, maintaining cell health, and delaying the aging process. Lycopene can most effectively scavenge free radicals in the body, maintain normal cell metabolism, and prevent aging. Therefore, the three also have a synergistic effect.

[0037] The anti-aging oral liquid prepared by this invention has a simple preparation method, the raw materials are widely available, the oral liquid has excellent antioxidant, anti-aging and anti-cancer effects, and the cost is low. It is easy to realize industrial production and has broad application prospects. Detailed Implementation

[0038] The technical solutions in the embodiments of the present invention will be clearly and completely described below. 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 skilled in the art without creative effort are within the scope of protection of the present invention.

[0039] Papain, 100,000 U / g, and bromelain, 100,000 U / g, were purchased from Nanning Dongheng Huadao Biotechnology Co., Ltd.

[0040] Trypsin, 250,000 U / g, purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0041] Egg yolks and egg whites were purchased from Guangzhou Baiyunshan Hanfang Modern Pharmaceutical Co., Ltd., with a protein content of 90-91%.

[0042] The Ganoderma lucidum seeds were purchased from Shandong Guanxian Kangrui Ganoderma lucidum Technology Co., Ltd., with a survival rate of >95% and a germination rate of >96%.

[0043] Example 1

[0044] This embodiment provides a method for preparing an anti-aging oral liquid, specifically including the following steps:

[0045] S1. Preparation of collagen peptide-copper complex: 100 parts by weight of cod skin were descaled, washed, minced, added to 500 parts by weight of water, 2 parts by weight of protease were added, and enzymatic hydrolysis was carried out at 40℃ for 2 hours. 1 part by weight of copper chloride was added, and the mixture was stirred at 35℃ for 0.5 hours. The mixture was filtered, and the filtrate was dialyzed through a dialysis bag with a pore size of 3 kDa for 3 hours. The mixture was then freeze-dried to obtain collagen peptide-copper complex.

[0046] The protease is a mixture of papain and bromelain in a mass ratio of 7:3;

[0047] S2. Two-way fermentation of traditional Chinese medicine: 1 part by weight of ginseng, 7 parts by weight of turmeric, 3 parts by weight of astragalus and 3 parts by weight of ginger are washed, crushed and mixed to obtain traditional Chinese medicine powder. 500 parts by weight of water, 7 parts by weight of glucose, 5 parts by weight of peptone and 2 parts by weight of sodium chloride are added and stirred evenly. The mixture is sterilized and inoculated with Ganoderma lucidum seed liquid at an inoculation amount of 8% and a mass fraction of 10%. Fermentation is carried out at 28℃ and 100r / min for 5 days. The mixture is then filtered, and the filtrate is the fermentation product.

[0048] S3. Preparation of polysaccharides from traditional Chinese medicine: Add ethanol to the fermentation product obtained in step S2 until the ethanol content of the system is 70 wt%, precipitate for 3 h, filter, dry the solid to obtain polysaccharides from traditional Chinese medicine, freeze-dry the filtrate to obtain the active component;

[0049] S4. Preparation of egg yolk protein-collagen peptide-copper composition: 10 parts by weight of egg yolk protein were added to 200 parts by weight of water, 7 parts by weight of the collagen peptide-copper complex obtained in step S1 were added, the pH of the solution was adjusted to 7.5, 0.3 parts by weight of trypsin was added, the mixture was heated to 35°C, stirred and hydrolyzed for 1 hour, the pH of the solution was adjusted to neutral, the enzyme was inactivated, the mixture was filtered, and the filtrate was freeze-dried to obtain the egg yolk protein-collagen peptide-copper composition;

[0050] S5. Preparation of active ingredient encapsulation solution: 15 parts by weight of the egg yolk protein-collagen peptide-copper composition obtained in step S4 were added to 200 parts by weight of water, stirred and mixed for 1 hour, hydrated overnight at 3°C, the pH of the solution was adjusted to 10, 3 parts by weight of the active component obtained in step S3 were added, stirred and mixed for 30 minutes, the pH of the solution was adjusted to 7, centrifuged, and the solid was removed to obtain the active ingredient encapsulation solution.

[0051] S6. Preparation of additive: Mix 3 parts by weight of resveratrol, 1 part by weight of taurine and 2 parts by weight of lycopene for 10 minutes to obtain the additive.

[0052] S7. Preparation of anti-aging oral liquid: Add 5 parts by weight of the Chinese herbal polysaccharide obtained in step S3 and 1 part by weight of the additive obtained in step S6 to 100 parts by weight of the active ingredient encapsulation solution obtained in step S5, homogenize at 10000 r / min for 5 min, filter, and obtain anti-aging oral liquid.

[0053] Example 2

[0054] This embodiment provides a method for preparing an anti-aging oral liquid, specifically including the following steps:

[0055] S1. Preparation of collagen peptide-copper complex: 100 parts by weight of cod skin were descaled, washed, minced, added to 500 parts by weight of water, 4 parts by weight of protease were added, and enzymatic hydrolysis was carried out at 50°C for 4 hours. 2 parts by weight of copper sulfate were added, and the mixture was stirred at 45°C for 1 hour. The mixture was filtered, and the filtrate was dialyzed through a dialysis bag with a pore size of 5 kDa for 5 hours. The mixture was then freeze-dried to obtain collagen peptide-copper complex.

[0056] The protease is a mixture of papain and bromelain in a mass ratio of 10:5;

[0057] S2. Two-way fermentation of traditional Chinese medicine: 2 parts by weight of ginseng, 10 parts by weight of turmeric, 5 parts by weight of astragalus and 5 parts by weight of ginger were washed, crushed and mixed to obtain traditional Chinese medicine powder. 500 parts by weight of water, 12 parts by weight of glucose, 7 parts by weight of peptone and 3 parts by weight of sodium chloride were added and stirred evenly. The mixture was sterilized and inoculated with Ganoderma lucidum seed liquid at an inoculation amount of 10% and a mass fraction of 8% of the Ganoderma lucidum seed liquid. Fermentation was carried out at 32℃ and 150r / min for 7 days. The mixture was then filtered and the filtrate was the fermentation product.

[0058] S3. Preparation of polysaccharides from traditional Chinese medicine: Add ethanol to the fermentation product obtained in step S2 until the ethanol content of the system is 80 wt%, precipitate for 5 h, filter, dry the solid to obtain polysaccharides from traditional Chinese medicine, freeze-dry the filtrate to obtain the active component;

[0059] S4. Preparation of egg yolk protein-collagen peptide-copper composition: 12 parts by weight of egg yolk protein were added to 200 parts by weight of water, 10 parts by weight of the collagen peptide-copper complex obtained in step S1 were added, the pH of the solution was adjusted to 8.5, 0.5 parts by weight of trypsin was added, the mixture was heated to 45°C, stirred and hydrolyzed for 3 hours, the pH of the solution was adjusted to neutral, the enzyme was inactivated, the mixture was filtered, and the filtrate was freeze-dried to obtain the egg yolk protein-collagen peptide-copper composition;

[0060] S5. Preparation of active ingredient encapsulation solution: 22 parts by weight of the egg yolk protein-collagen peptide-copper composition obtained in step S4 were added to 200 parts by weight of water, stirred and mixed for 1 hour, hydrated overnight at 5°C, the pH of the solution was adjusted to 12, 5 parts by weight of the active component obtained in step S3 were added, stirred and mixed for 30 minutes, the pH of the solution was adjusted to 7, centrifuged, and the solid was removed to obtain the active ingredient encapsulation solution.

[0061] S6. Preparation of additive: Mix 5 parts by weight of resveratrol, 2 parts by weight of taurine and 4 parts by weight of lycopene for 10 minutes to obtain the additive.

[0062] S7. Preparation of anti-aging oral liquid: Add 7 parts by weight of the Chinese herbal polysaccharide obtained in step S3 and 3 parts by weight of the additive obtained in step S6 to 100 parts by weight of the active ingredient encapsulation solution obtained in step S5, homogenize at 12000 r / min for 10 min, filter, and obtain anti-aging oral liquid.

[0063] Example 3

[0064] This embodiment provides a method for preparing an anti-aging oral liquid, specifically including the following steps:

[0065] S1. Preparation of collagen peptide-copper complex: 100 parts by weight of cod skin were descaled, washed, minced, added to 500 parts by weight of water, 3 parts by weight of protease were added, and enzymatic hydrolysis was carried out at 45°C for 3 hours. 1.5 parts by weight of copper nitrate were added, and the mixture was stirred at 40°C for 1 hour. The mixture was filtered, and the filtrate was dialyzed through a dialysis bag with a pore size of 4 kDa for 4 hours. The mixture was then freeze-dried to obtain collagen peptide-copper complex.

[0066] The protease is a mixture of papain and bromelain in a mass ratio of 8:4;

[0067] S2. Two-way fermentation of traditional Chinese medicine: 1.5 parts by weight of ginseng, 8.5 parts by weight of turmeric, 4 parts by weight of astragalus, and 4 parts by weight of ginger were washed, crushed, and mixed to obtain traditional Chinese medicine powder. The powder was added to 500 parts by weight of water, along with 10 parts by weight of glucose, 6 parts by weight of peptone, and 2.5 parts by weight of sodium chloride. The mixture was stirred and mixed evenly, sterilized, and inoculated with Ganoderma lucidum seed liquid at an inoculation amount of 9%. The mass fraction of the Ganoderma lucidum seed liquid was 9%. The mixture was fermented at 30℃ and 120 r / min for 6 days. The mixture was then filtered, and the filtrate was the fermentation product.

[0068] S3. Preparation of polysaccharides from traditional Chinese medicine: Add ethanol to the fermentation product obtained in step S2 until the ethanol content of the system is 75 wt%, precipitate for 4 h, filter, dry the solid to obtain polysaccharides from traditional Chinese medicine, freeze-dry the filtrate to obtain the active component;

[0069] S4. Preparation of egg yolk protein-collagen peptide-copper composition: 11 parts by weight of egg yolk protein were added to 200 parts by weight of water, 8 parts by weight of the collagen peptide-copper complex obtained in step S1 were added, the pH of the solution was adjusted to 8, 0.4 parts by weight of trypsin was added, the mixture was heated to 40°C, stirred and hydrolyzed for 2 hours, the pH of the solution was adjusted to neutral, the enzyme was inactivated, the mixture was filtered, and the filtrate was freeze-dried to obtain the egg yolk protein-collagen peptide-copper composition;

[0070] S5. Preparation of active ingredient encapsulation solution: Add 20 parts by weight of the egg yolk protein-collagen peptide-copper composition obtained in step S4 to 200 parts by weight of water, stir and mix for 1 hour, hydrate overnight at 4°C, adjust the pH of the solution to 11, add 4 parts by weight of the active component obtained in step S3, stir and mix for 30 minutes, adjust the pH of the solution to 7, centrifuge, remove the solid, and obtain the active ingredient encapsulation solution.

[0071] S6. Preparation of additive: Mix 4 parts by weight of resveratrol, 1.5 parts by weight of taurine and 3 parts by weight of lycopene for 10 minutes to obtain the additive.

[0072] S7. Preparation of anti-aging oral liquid: Add 6 parts by weight of the Chinese herbal polysaccharide obtained in step S3 and 2 parts by weight of the additive obtained in step S6 to 100 parts by weight of the active ingredient encapsulation solution obtained in step S5, homogenize at 11000 r / min for 7 min, filter, and obtain anti-aging oral liquid.

[0073] Example 4

[0074] The difference from Example 3 is that the protease is papain.

[0075] Example 5

[0076] The difference from Example 3 is that the protease used is bromelain.

[0077] Comparative Example 1

[0078] The difference from Example 3 is that no protease was added in step S1.

[0079] Specifically as follows:

[0080] S1. Preparation of collagen-copper complex: 100 parts by weight of cod skin were descaled, washed, minced, added to 500 parts by weight of water, and 1.5 parts by weight of copper nitrate were added. The mixture was stirred at 40°C for 1 hour, filtered, and the filtrate was dialyzed through a dialysis bag with a pore size of 4 kDa for 4 hours. The mixture was then freeze-dried to obtain collagen-copper complex.

[0081] Comparative Example 2

[0082] The difference from Example 3 is that copper nitrate was not added in step S1.

[0083] Specifically as follows:

[0084] S1. Preparation of collagen peptides: 100 parts by weight of cod skin were descaled, washed, minced, added to water, and 3 parts by weight of protease were added. The mixture was enzymatically hydrolyzed at 45°C for 3 hours, filtered, and the filtrate was dialyzed through a dialysis bag with a pore size of 4 kDa for 4 hours. The mixture was then freeze-dried to obtain collagen peptides.

[0085] Comparative Example 3

[0086] The difference from Example 3 is that no egg yolk protein was added in step S4.

[0087] Specifically as follows:

[0088] S4. Preparation of collagen peptide-copper composition: 19 parts by weight of the collagen peptide-copper complex obtained in step S1 were added to 200 parts by weight of water, the pH of the solution was adjusted to 8, 0.4 parts by weight of trypsin was added, the mixture was heated to 40°C, stirred and hydrolyzed for 2 hours, the pH of the solution was adjusted to neutral, the enzyme was inactivated, the mixture was filtered, and the filtrate was freeze-dried to obtain the collagen peptide-copper composition.

[0089] Comparative Example 4

[0090] The difference from Example 3 is that collagen peptide-copper complex was not added in step S4.

[0091] Specifically as follows:

[0092] S4. Preparation of egg yolk protein peptide composition: 19 parts by weight of egg yolk protein were added to 200 parts by weight of water, the pH of the solution was adjusted to 8, 0.4 parts by weight of trypsin was added, the mixture was heated to 40°C, stirred and hydrolyzed for 2 hours, the pH of the solution was adjusted to neutral, the enzyme was inactivated, the mixture was filtered, and the filtrate was freeze-dried to obtain the egg yolk protein peptide composition.

[0093] Comparative Example 5

[0094] The difference from Example 3 is that step S4 was not performed.

[0095] Specifically as follows:

[0096] S1. Preparation of collagen peptide-copper complex: 100 parts by weight of cod skin were descaled, washed, minced, added to 500 parts by weight of water, 3 parts by weight of protease were added, and enzymatic hydrolysis was carried out at 45°C for 3 hours. 1.5 parts by weight of copper nitrate were added, and the mixture was stirred at 40°C for 1 hour. The mixture was filtered, and the filtrate was dialyzed through a dialysis bag with a pore size of 4 kDa for 4 hours. The mixture was then freeze-dried to obtain collagen peptide-copper complex.

[0097] The protease is a mixture of papain and bromelain in a mass ratio of 8:4;

[0098] S2. Two-way fermentation of traditional Chinese medicine: 1.5 parts by weight of ginseng, 8.5 parts by weight of turmeric, 4 parts by weight of astragalus, and 4 parts by weight of ginger were washed, crushed, and mixed to obtain traditional Chinese medicine powder. The powder was added to 500 parts by weight of water, along with 10 parts by weight of glucose, 6 parts by weight of peptone, and 2.5 parts by weight of sodium chloride. The mixture was stirred and mixed evenly, sterilized, and inoculated with Ganoderma lucidum seed liquid at an inoculation amount of 9%. The mass fraction of the Ganoderma lucidum seed liquid was 9%. The mixture was fermented at 30℃ and 120 r / min for 6 days. The mixture was then filtered, and the filtrate was the fermentation product.

[0099] S3. Preparation of polysaccharides from traditional Chinese medicine: Add ethanol to the fermentation product obtained in step S2 until the ethanol content of the system is 75 wt%, precipitate for 4 h, filter, dry the solid to obtain polysaccharides from traditional Chinese medicine, freeze-dry the filtrate to obtain the active component;

[0100] S4. Preparation of active ingredient solution: Add 4 parts by weight of the active component obtained in step S3 to 200 parts by weight of water to obtain an active ingredient solution;

[0101] S5. Preparation of additive: Mix 4 parts by weight of resveratrol, 1.5 parts by weight of taurine and 3 parts by weight of lycopene for 10 minutes to obtain the additive.

[0102] S6. Preparation of anti-aging oral liquid: Add 6 parts by weight of the Chinese herbal polysaccharide obtained in step S3 and 2 parts by weight of the additive obtained in step S6 to 100 parts by weight of the active ingredient solution obtained in step S4, homogenize at 11000 r / min for 7 min, filter, and obtain anti-aging oral liquid.

[0103] Comparative Example 6

[0104] The difference from Example 3 is that no active ingredient was added in step S5.

[0105] Specifically as follows:

[0106] S5. Preparation of protein peptide solution: 20 parts by weight of the egg yolk protein-collagen peptide-copper composition obtained in step S4 were added to 200 parts by weight of water, stirred and mixed for 1 hour, and hydrated overnight at 4°C to obtain protein peptide solution.

[0107] Comparative Example 7

[0108] The difference from Example 3 is that no traditional Chinese medicine polysaccharides were added in step S7.

[0109] Specifically as follows:

[0110] S7. Preparation of anti-aging oral liquid: Add 2 parts by weight of the additive obtained in step S6 to 100 parts by weight of the active ingredient encapsulation solution obtained in step S5, homogenize at 11000 r / min for 7 min, filter, and obtain anti-aging oral liquid.

[0111] Comparative Example 8

[0112] The difference from Example 3 is that no additives were added in step S7.

[0113] Specifically as follows:

[0114] S7. Preparation of anti-aging oral liquid: Add 6 parts by weight of the Chinese herbal polysaccharide obtained in step S3 to 100 parts by weight of the active ingredient encapsulation solution obtained in step S5, homogenize at 11000 r / min for 7 min, filter, and obtain anti-aging oral liquid.

[0115] Test Example 1 Antioxidant Effect

[0116] The anti-aging oral liquids prepared in Examples 1-5 and Comparative Examples 1-8 were diluted 100 times to prepare sample solutions.

[0117] 1. DPPH free radical scavenging rate determination

[0118] Each sample solution, anhydrous ethanol, and 0.2 mmol / L DPPH solution were prepared according to Table 1. The mixture was reacted at 25°C in the dark for 30 min, centrifuged, and the absorbance was measured at 517 nm. Each sample was measured in triplicate, and the average value was taken. The concentration of the vitamin C solution was 10 mg / L.

[0119] Table 1

[0120] reagents Test sample (mL) Anhydrous ethanol (mL) DPPH solution (mL) Sample group 1 0 1 Vitamin C group 1 0 1 negative control group 0 1 1 Blank control group 1 1 0

[0121] DPPH free radical scavenging rate (%) = (A2 + A3 - A1) / A2 × 100%

[0122] In the formula, A2: absorbance of the negative control group; A3: absorbance of the blank control group; A1: absorbance of the sample group / positive control group.

[0123] 2. Determination of ABTS free radical scavenging rate

[0124] Prepare 7.4 mmol / L ABTS stock solution and 2.6 mmol / L K₂S₂O₈ stock solution. Take 2 mL of each and mix them at room temperature, protected from light, and store for 15 h. Before the experiment, dilute with PBS buffer solution (pH 4.0) to obtain an absorbance of 0.7 ± 0.05 at 734 nm, thus obtaining the ABTS working solution. Prepare the sample solution, 50 wt% ethanol solution, and ABTS working solution according to Table 2. Incubate at room temperature, protected from light, for 8 min, and measure the absorbance at 734 nm. Perform triplicate measurements for each sample and take the average value. The concentration of vitamin C solution is 10 mg / L.

[0125] Table 2

[0126] reagents Test sample (mL) 50wt% ethanol (mL) ABTS solution (mL) Sample group 0.5 0 0.5 Vitamin C group 0.5 0 0.5 negative control group 0 0.5 0.5 Blank control group 0.5 0.5 0

[0127] ABTS free radical scavenging rate (%) = [1-(A i -A0) / A j ]×100%

[0128] In the formula, A i : Absorbance of sample group / positive control group; A j A0: Absorbance of the blank control group; A0: Absorbance of the negative control group.

[0129] The results are shown in Table 3.

[0130] Table 3

[0131]

[0132]

[0133] As can be seen from the table above, the anti-aging oral liquids prepared in Examples 1-3 of this invention have good antioxidant effects.

[0134] Test Example 2

[0135] Male Kunming (KM) mice weighing 18-22g were selected and, after 7 days of acclimatization, randomly divided into 16 groups: a control group, a positive control group, a model group, Examples 1-5, and Comparative Examples 1-8. Except for the control group, which received subcutaneous injections of physiological saline (0.5 mL / kg), all other groups received subcutaneous injections of 500 mg / kg of 0.9% sodium chloride D-galactose solution, along with gavage intervention. The positive control group received 17 mg / kg of vitamin C via gavage, while Examples 1-5 and Comparative Examples 1-8 received 2 mL / kg / day of the corresponding prepared anti-aging oral solution via gavage. The control and model groups received an equal volume of physiological saline. Treatment was administered once daily for 14 days. After the administration period, each group of mice underwent a water maze test and a platform jumping test.

[0136] 1. Water Maze Experiment

[0137] Before starting, mice were trained to locate the platform, with two successful attempts to climb it considered sufficient. On the third day, a random entry point was chosen, and the mice were placed in the pool. The time taken to find the platform and the escape latency were recorded. If the platform was not found within 90 seconds, the latency was recorded as 90 seconds.

[0138] 2. Diving platform experiment

[0139] On the first day, mice were placed in a box to acclimatize to the environment, then placed on a platform. The time it took for the mouse to jump off the platform for the first time was recorded as the latency period. On the second day, mice were placed on the platform. When they jumped off, electrical stimulation was immediately applied. After 3-4 seconds, the mice were returned to their cages. The formal experiment began on the third day. The latency period and the number of platform jumps within 5 minutes (i.e., the number of errors) were recorded. If a mouse did not jump off the platform within 5 minutes, the latency period was recorded as 300 seconds.

[0140] The results are shown in Table 4.

[0141] Table 4

[0142]

[0143] Note: * indicates P < 0.05 compared to the control group; # indicates P < 0.05 compared to the model group.

[0144] As shown in the table above, the anti-aging oral liquids prepared in Examples 1-3 of this invention can significantly improve the memory of D-galactose-induced aging model mice.

[0145] 3. Antioxidant index testing

[0146] After the experiment, mice were fasted for 12 hours, blood was collected from their eyes, allowed to stand for 2 hours, centrifuged, and the supernatant was collected and stored at 4°C for later use. The levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in mouse serum were determined according to the kit instructions. The results are shown in Table 5.

[0147] Table 5

[0148] Group MDA content (nmol / mL) SOD content (U / mL) Blank group 12.92±1.45 624.28±10.25 Model group 35.62±4.18* 481.19±6.79* positive group 49.25±3.25# 282.56±5.10# Example 1 16.77±1.24# 592.61±11.18# Example 2 16.82±1.45# 590.28±12.35# Example 3 16.05±1.19# 594.07±9.28# Example 4 17.21±1.82 580.26±8.26 Example 5 17.19±1.48 582.19±9.47 Comparative Example 1 18.24±2.09 571.29±10.11 Comparative Example 2 18.04±1.48 577.21±9.19 Comparative Example 3 19.21±1.26 561.18±10.38 Comparative Example 4 21.11±1.34 552.26±9.27 Comparative Example 5 23.46±1.69 543.38±8.26 Comparative Example 6 29.58±1.46 500.19±7.58 Comparative Example 7 25.04±1.72 530.59±6.82 Comparative Example 8 28.49±1.64 517.28±4.96

[0149] Note: * indicates P < 0.05 compared to the control group; # indicates P < 0.05 compared to the model group.

[0150] As shown in the table above, the anti-aging oral liquids prepared in Examples 1-3 of this invention can significantly increase the content of superoxide dismutase and decrease the content of malondialdehyde in mice.

[0151] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing an anti-aging oral liquid, characterized in that, Ginseng, turmeric, astragalus, and ginger were fermented in a two-way manner using Ganoderma lucidum seed liquid. The product was precipitated with alcohol to obtain traditional Chinese medicine polysaccharides, and then freeze-dried to obtain the active components. Cod skin was extracted to obtain a collagen peptide-copper complex, which was mixed with egg yolk protein and enzymatically hydrolyzed to obtain an egg yolk protein-collagen peptide-copper composition. This composition was added to water and encapsulated with the active components to obtain an active ingredient encapsulation solution. Traditional Chinese medicine polysaccharides, resveratrol, taurine, and lycopene were added, homogenized, and filtered to obtain an anti-aging oral liquid.

2. The preparation method according to claim 1, characterized in that, Includes the following steps: S1. Preparation of collagen peptide-copper complex: Cod skin was scaled, washed, minced, added to water, enzymatically hydrolyzed with protease, copper salt was added, the reaction was stirred, filtered, the filtrate was dialyzed, and freeze-dried to obtain collagen peptide-copper complex. S2. Two-way fermentation of traditional Chinese medicine: Ginseng, turmeric, astragalus and ginger are washed, crushed and mixed to obtain traditional Chinese medicine powder. Add water, glucose, peptone and sodium chloride, stir and mix evenly, sterilize, inoculate with Ganoderma lucidum seed liquid, ferment and culture, filter, and the filtrate is the fermentation product. S3. Preparation of polysaccharides from traditional Chinese medicine: Ethanol is added to the fermentation product obtained in step S2 for precipitation, filtered, and the solid is dried to obtain polysaccharides from traditional Chinese medicine. The filtrate is freeze-dried to obtain the active component. S4. Preparation of egg yolk protein-collagen peptide-copper composition: Add egg yolk protein to water, add the collagen peptide-copper complex obtained in step S1, adjust the pH of the solution to alkaline, add trypsin, heat to hydrolyze, adjust the pH of the solution to neutral, inactivate the enzyme, filter, freeze-dry the filtrate to obtain egg yolk protein-collagen peptide-copper composition. S5. Preparation of active ingredient encapsulation solution: Add the egg yolk protein-collagen peptide-copper composition obtained in step S4 to water, stir and mix, hydrate overnight, adjust the pH of the solution to alkaline, add the active component obtained in step S3, stir and mix, adjust the pH of the solution to neutral, centrifuge, remove solids, and obtain active ingredient encapsulation solution. S6. Preparation of additives: Resveratrol, taurine and lycopene are mixed evenly to prepare additives; S7. Preparation of anti-aging oral liquid: The traditional Chinese medicine polysaccharide obtained in step S3 and the additive obtained in step S6 are added to the active ingredient encapsulation solution obtained in step S5, homogenized, filtered, and the anti-aging oral liquid is obtained.

3. The preparation method according to claim 2, characterized in that, The protease mentioned in step S1 is selected from at least one of trypsin, pepsin, papain, neutral protease, bromelain, alkaline protease, and fig protease. Preferably, it is a mixture of papain and bromelain in a mass ratio of 7-10:3-5. The enzymatic hydrolysis temperature is 40-50℃ and the time is 2-4 hours. The mass ratio of cod skin, protease, and copper salt is 100:2-4:1-2. The copper salt is selected from at least one of copper chloride, copper sulfate, and copper nitrate. The dialysis is performed using a dialysis bag with a pore size of 3-5 kDa and the dialysis time is 3-5 hours.

4. The preparation method according to claim 2, characterized in that, In step S2, the mass ratio of ginseng, turmeric, astragalus, ginger, glucose, peptone, and sodium chloride is 1-2:7-10:3-5:3-5:7-12:5-7:2-3. The inoculation amount of the Ganoderma lucidum seed liquid is 8-10%, the mass fraction of the Ganoderma lucidum seed liquid is 8-10%, and the fermentation culture conditions are 28-32℃, 100-150r / min, and culture for 5-7 days.

5. The preparation method according to claim 2, characterized in that, In step S3, ethanol is added until the ethanol content in the system is 70-80 wt%, and the precipitation time is 3-5 h.

6. The preparation method according to claim 2, characterized in that, In step S4, the mass ratio of egg yolk protein, collagen peptide-copper complex, and trypsin is 10-12:7-10:0.3-0.5, the pH of the solution is adjusted to be alkaline within the range of 7.5-8.5, and the temperature for hydrolysis is 35-45℃ for 1-3 hours.

7. The preparation method according to claim 2, characterized in that, In step S5, the mass ratio of the egg yolk protein-collagen peptide-copper composition and the active component is 15-22:3-5, the overnight hydration temperature is 3-5°C, and the pH of the solution is adjusted to be alkaline within the range of 10-12.

8. The preparation method according to claim 2, characterized in that, In step S6, the mass ratio of resveratrol, taurine, and lycopene is 3-5:1-2:2-4; in step S7, the mass ratio of traditional Chinese medicine polysaccharides, additives, and active ingredient encapsulation solution is 5-7:1-3:100; the homogenization speed is 10000-12000 r / min, and the time is 5-10 min.

9. The preparation method according to claim 2, characterized in that, Specifically, the following steps are included: S1. Preparation of collagen peptide-copper complex: 100 parts by weight of cod skin were descaled, washed, minced, added to 500 parts by weight of water, 2-4 parts by weight of protease were added, and enzymatic hydrolysis was carried out at 40-50℃ for 2-4 hours. 1-2 parts by weight of copper salt were added, and the mixture was stirred at 35-45℃ for 0.5-1 hours. The mixture was filtered, and the filtrate was dialyzed through a dialysis bag with a pore size of 3-5 kDa for 3-5 hours. The mixture was then freeze-dried to obtain collagen peptide-copper complex. The protease is a mixture of papain and bromelain in a mass ratio of 7-10:3-5; S2. Two-way fermentation of traditional Chinese medicine: Wash and crush 1-2 parts by weight of ginseng, 7-10 parts by weight of turmeric, 3-5 parts by weight of astragalus, and 3-5 parts by weight of ginger, mix them to obtain traditional Chinese medicine powder, add 500 parts by weight of water, add 7-12 parts by weight of glucose, 5-7 parts by weight of peptone, and 2-3 parts by weight of sodium chloride, stir and mix evenly, sterilize, inoculate with Ganoderma lucidum seed liquid, the inoculation amount is 8-10%, the mass fraction of the Ganoderma lucidum seed liquid is 8-10%, 28-32℃, 100-150r / min, ferment and culture for 5-7 days, filter, the filtrate is the fermentation product; S3. Preparation of polysaccharides from traditional Chinese medicine: Add ethanol to the fermentation product obtained in step S2 until the ethanol content of the system is 70-80 wt%, precipitate for 3-5 hours, filter, dry the solid to obtain polysaccharides from traditional Chinese medicine, freeze-dry the filtrate to obtain the active component; S4. Preparation of egg yolk protein-collagen peptide-copper composition: 10-12 parts by weight of egg yolk protein were added to 200 parts by weight of water, 7-10 parts by weight of the collagen peptide-copper complex obtained in step S1 were added, the pH of the solution was adjusted to 7.5-8.5, 0.3-0.5 parts by weight of trypsin were added, the mixture was heated to 35-45℃, stirred and hydrolyzed for 1-3 hours, the pH of the solution was adjusted to neutral, the enzyme was inactivated, the mixture was filtered, and the filtrate was freeze-dried to obtain the egg yolk protein-collagen peptide-copper composition; S5. Preparation of active ingredient encapsulation solution: Add 15-22 parts by weight of the egg yolk protein-collagen peptide-copper composition obtained in step S4 to 200 parts by weight of water, stir and mix, hydrate overnight at 3-5℃, adjust the pH of the solution to 10-12, add 3-5 parts by weight of the active component obtained in step S3, stir and mix, adjust the pH of the solution to 7, centrifuge, remove solids, and obtain active ingredient encapsulation solution; S6. Preparation of additive: Mix 3-5 parts by weight of resveratrol, 1-2 parts by weight of taurine and 2-4 parts by weight of lycopene evenly to prepare the additive. S7. Preparation of anti-aging oral liquid: Add 5-7 parts by weight of the Chinese herbal polysaccharide obtained in step S3 and 1-3 parts by weight of the additive obtained in step S6 to 100 parts by weight of the active ingredient encapsulation solution obtained in step S5, homogenize at 10000-12000 r / min for 5-10 min, filter, and obtain the anti-aging oral liquid.

10. An anti-aging oral liquid prepared by any one of claims 1-9.

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