Compound peptide beverage capable of dispelling effects of alcohol and protecting liver as well as preparation method and application of compound peptide beverage

Abstract

The invention provides a compound peptide beverage capable of dispelling effects of alcohol and protecting liver as well as a preparation method and application thereof. The compound peptide beverage is prepared from hovenia dulcis thunb oligopeptide, walnut peptide, antioxidant peptide, energy release auxiliary peptide, ethanol dehydrogenase activating peptide, silibinin-serine chelate, an electrolyte poising agent, alginic acid oligosaccharide and purified water. Wherein the antioxidant peptide is prepared from cordyceps militaris, sea-buckthorn and sunflower disc through mixed enzymolysis, the energy release auxiliary peptide is prepared from carnosine and arginine succinic acid peptide through microbial fermentation, and the ethanol dehydrogenase activating peptide is prepared from marine shellfish collagen through fermentation. The composite peptide beverage shows excellent functions of dispelling the effects of alcohol and protecting the liver, and can be used for developing medicinal products or functional foods for dispelling the effects of alcohol and protecting the liver. Through precise compatibility and an innovative process technology, the product constructs a unique solution for dispelling the effects of alcohol and protecting the liver, and has outstanding performance in the aspects of improving the alcohol metabolism efficiency and relieving liver injury.

Description

Technical Field

[0001] This invention relates to the field of peptide technology, specifically to a compound peptide beverage for relieving hangovers and protecting the liver, its preparation method, and its application. Background Technology

[0002] In modern life, alcohol consumption has become a common way to socialize and release stress, but the damage to the liver caused by excessive drinking cannot be ignored. Therefore, liver protection and hangover relief are essential. The core of liver protection and hangover relief lies in accelerating ethanol metabolism, reducing acetaldehyde accumulation, and repairing liver damage. Traditional hangover relief products often suffer from slow effects and significant side effects. Peptide drinks, due to their high absorption rate and low side effects, are a better choice. Although peptide drinks on the market have shown potential in the field of liver protection and hangover relief, most products still have shortcomings. Existing peptide drinks have relatively limited functions; most products only focus on accelerating ethanol metabolism while neglecting the repair of alcoholic liver damage, resulting in poor long-term protective effects. Summary of the Invention

[0003] In view of this, the present invention proposes a hangover-relieving and liver-protecting compound peptide beverage, its preparation method and application, to solve the above problems.

[0004] The technical solution of this invention is implemented as follows: A complex peptide beverage for hangover relief and liver protection comprises the following ingredients by weight: 10-18 parts of Hovenia dulcis oligopeptides, 5-10 parts of walnut peptides, 18-35 parts of antioxidant peptides, 8-15 parts of energy-releasing co-peptides, 6-8 parts of alcohol dehydrogenase activating peptides, 0.5-2.0 parts of silymarin-serine chelate, 0.3-0.8 parts of electrolyte balancer, 1-3 parts of alginate oligosaccharides, and 200-300 parts of purified water. The antioxidant peptides are prepared by enzymatic hydrolysis of a mixture of Cordyceps militaris, Hippophae rhamnoides, and sunflower disc in a mass ratio of (14-25):(8-10):(3-5). The energy-releasing co-peptides are prepared by microbial fermentation of carnosine and arginine succinate peptides in a volume ratio of 1-3:0.2. The alcohol dehydrogenase activating peptides are prepared by fermentation of marine shellfish collagen.

[0005] Furthermore, the Hovenia dulcis oligopeptides were prepared by the following method: Fresh, disease-free, and pollution-free Hovenia dulcis seeds were selected, dried, and then pulverized into fine powder. Pure water was added at a ratio of g / mL of 1:1-3 and stirred evenly. A compound enzyme was added and hydrolyzed at 38-45℃ and pH 7.0-9.0 for 2-4 hours. After hydrolysis, the enzyme was inactivated at 80-90℃ for 10-15 minutes. The mixture was then centrifuged, and the supernatant was subjected to a three-stage gradient ultrafiltration. First, a 50 kDa membrane was used for ultrafiltration. The resulting filtrate ① was then ultrafiltered using a 10 kDa membrane, and the resulting filtrate ② was ultrafiltered using a 1 kDa membrane to obtain Hovenia dulcis oligopeptides with a molecular weight <1000 Da. The amount of compound enzyme added is 1.5-3.0% of the mass of Hovenia dulcis powder. The compound enzyme is composed of Bacillus subtilis protease and alkaline protease in a mass ratio of 1:2-5. The enzyme activities of Bacillus subtilis protease and alkaline protease are ≥3000 IU / g.

[0006] Furthermore, walnut peptides are prepared by the following method: Fresh, unspoiled walnuts are selected, the shells are removed to obtain walnut kernels, which are then crushed and passed through a 60-100 mesh sieve to obtain walnut meal powder. First, a hexane-ethanol solution with a volume ratio of 1.5-2.0:1 is used to defatt the walnut meal powder at 45-55℃ with stirring for 25-35 minutes. Then, the defatted meal powder is filtered out, rinsed with purified water, and then soaked in a 0.1-0.3% sodium sulfite aqueous solution for 30-50 minutes. Next, water is added, and the pH is adjusted to 7.0-9.0 with a dilute alkaline solution. Finally, the mixture is heated to 50°C. Ultrasonic extraction was performed at 0-60℃, followed by centrifugation to obtain supernatant a. The filter residue was subjected to the same ultrasonic extraction and centrifugation process to obtain supernatant b. Supernatants a and b were combined and concentrated to obtain a concentrate. Alkaline protease was added to the concentrate for the first enzymatic hydrolysis, followed by neutral protease for the second enzymatic hydrolysis. After the enzymatic hydrolysis, the enzyme was inactivated at 80-90℃ for 10-15 min to obtain the hydrolysate. The hydrolysate was precipitated, and the supernatant was collected. The supernatant was centrifuged to obtain supernatant c. Supernatant c was subjected to two-stage gradient ultrafiltration. First, a 30 kDa membrane was used for ultrafiltration, and the resulting filtrate (③) was then ultrafiltered using a 3 kDa membrane to obtain walnut peptides. The enzyme addition for the first enzymatic hydrolysis was 1.5-2.0% of the concentrate volume, the hydrolysis temperature was 30-60℃, the pH was 8.0-9.0, and the hydrolysis time was 3-6 h. The amount of enzyme added for the second enzymatic hydrolysis is 0.5-1.0% of the volume of the concentrated liquid, the hydrolysis temperature is 28-38℃, the pH is 6.0-7.0, and the hydrolysis time is 2.5-3.5h.

[0007] Furthermore, the antioxidant peptides were prepared by the following method: Cordyceps militaris, Hippophae rhamnoides, and sunflower discs were mixed in proportion, washed, dried at 60-80℃ and pulverized into 50-80 mesh particles to obtain a mixed powder. Pure water was added to the mixed powder at a material-to-liquid ratio of 3-5:10 (g / mL) to obtain a mixed solution. The pH was adjusted to 7.0-9.0, and the solution was extracted at 90-100℃ for 50-70 minutes. After centrifugation, the supernatant was collected, and the pH was adjusted to 6.0-7.0. Bromelain, rhamnosidase, and aminopeptidase were added at a mass ratio of 1:(0.5-0.8):(0.1-0.3). Enzymatic hydrolysis was carried out at 40-60℃ for 2-4 hours. After hydrolysis, the enzymes were inactivated at 80-90℃ for 10-15 minutes to obtain the hydrolysate. The hydrolysate was centrifuged, and the supernatant was subjected to two-stage gradient ultrafiltration. First, a 10 kDa membrane was used for ultrafiltration, and the filtrate ④ was then subjected to two-stage gradient ultrafiltration. Ultrafiltration was performed using a kDa membrane, followed by conditioning with steam at 90-120℃ for 3-5 minutes to obtain antioxidant peptides.

[0008] Furthermore, the energy-releasing co-peptide was prepared by the following method: Carnosine and arginine succinate peptide were mixed in a volume ratio, and microorganisms were added and fermented at 25-37℃ for 24-72 h. The supernatant was then collected by centrifugation to obtain the energy-releasing co-peptide. The microorganisms consisted of Bacillus and Lactobacillus in a mass ratio of (3.5-8.2):(1.2-3.0), with a bacterial count ≥2.0 × 10⁻⁶. 8 CFU / g; the amino acid succinate peptide was prepared by the following method: L-arginine and succinic anhydride were mixed at a volume ratio of 1:1.0-1.5 and dissolved in phosphate buffer at pH 8.0-9.0. The mixture was stirred at 25-30℃ for 6-8 h. After the reaction, the pH was adjusted to 4.0-5.0 to precipitate the product. The precipitate was collected by centrifugation and washed with anhydrous ethanol at 4-8℃ to obtain the arginine succinate monomer. The obtained arginine succinate monomer was dissolved in Tris-HCl buffer at pH 7.0-7.5 to prepare a 90-110 mM solution. Glutaminase and 0.1 M sodium chloride solution were added to obtain the reaction solution. The mixture was stirred at 35-40℃ for 6-8 h. The polymerization process was monitored by SDS-PAGE. When the molecular weight reached 1-2 kDa, the temperature was raised to 80-90℃ and maintained for 10-15 min to terminate the reaction. The reaction solution was subjected to 2 Ultrafiltration was performed using a kDa membrane to obtain arginine succinate peptide; the amount of glutamine transaminase added was 2.0-3.0% of the mass of the reaction solution, and the enzyme activity of the glutamine transaminase was 200-400 IU / g.

[0009] Furthermore, the alcohol dehydrogenase activating peptide was prepared by the following method: Fresh marine shellfish were shelled, viscerated, and cleaned. The shellfish were then enzymatically hydrolyzed with papain at pH 6.5-7.5 and a temperature of 50-60℃ for 5-7 hours to obtain the hydrolysate. The hydrolysate was then inactivated with enzymes at 85-90℃ for 10-15 minutes. Subsequently, *Lactobacillus plantarum* was inoculated and fermented at pH 6.0-6.5, a temperature of 35-40℃, and a humidity of 75-85% for 60-84 hours, with aeration and stirring every 12 hours. After fermentation, the mixture was centrifuged, and the supernatant was ultrafiltered at 3 kDa. The filtrate was then separated using a Sephadex G-25 gel chromatography column with 0.05 M phosphate buffer as the mobile phase. Fractions with a retention time of 18-22 minutes were collected and further purified by reversed-phase high-performance liquid chromatography to obtain alcohol dehydrogenase activating peptides with molecular weights concentrated in the range of 500-1500 Da. The amount of papain added is 3-5% of the mass of the marine shellfish, with an enzyme activity ≥4000 IU / g; the amount of Lactobacillus plantarum added is 4-7% of the mass of the hydrolysate, with a Lactobacillus plantarum viable count ≥5.0×10⁻⁶. 8 CFU / g.

[0010] Furthermore, the silybin-serine chelate was prepared by the following method: silybin and L-serine were dissolved in 50 mM Tris-HCl buffer at a mass ratio of 1:1.1-1.5 at pH 7.5-8.5, and the mixture was stirred at 58-62℃ in the dark for 6-8 h. The reaction solution was then filtered through a 0.2-0.3 μm filter and subsequently subjected to anion exchange chromatography using a DEAE Sepharose FF column. The fraction with a retention time of 15-18 min was collected, and the collected fraction was placed in a 75-85% ethanol aqueous solution at 3-5℃ and allowed to stand for 10-15 h. The solid component was filtered and dried at 40-50℃ for 6-8 h, and then pulverized through a 100-150 mesh sieve to obtain the silybin-serine chelate.

[0011] Furthermore, the electrolyte balancer is any one or a combination of several of sodium chloride, potassium chloride, calcium lactate, sodium citrate, magnesium malate, and zinc gluconate.

[0012] Furthermore, a hangover-relieving and liver-protecting complex peptide beverage is prepared by the following method, including the following steps: S1. Mix silymarin-serine chelate, electrolyte balancer, and alginate oligosaccharide according to the weight parts, add purified water preheated to 40-50℃, place in a stirrer and stir at 400-800rpm for 15-25min to obtain solution ①.

[0013] S2. Add the Hovenia dulcis oligopeptide, walnut peptide, antioxidant peptide and energy release co-peptide to solution ① in sequence, stir at 60-100 rpm for 10-20 min, then slowly inject the alcohol dehydrogenase activating peptide, and adjust the pH to 6.8-7.2 with 8-12% citric acid solution or 4-6% sodium bicarbonate solution to obtain solution ②.

[0014] S3. Place solution ② into a homogenizer and homogenize it 2-3 times at 20-30MPa. Then perform UHT sterilization at 120-130℃ for 5-10 seconds. After that, fill the bottle to obtain the hangover relief and liver protection compound peptide beverage.

[0015] Further applications of compound peptide drinks include hangover relief and liver protection health products or drugs.

[0016] Compared with the prior art, the present invention has the following beneficial effects: This invention utilizes antioxidant peptides and alginate oligosaccharides to scavenge free radicals and toxins, reducing toxin entry into the liver and constructing a preventative system. It also employs silymarin-serine chelates to target and repair membrane structures, stabilizing organelle function and constructing a repair system. Furthermore, it provides amino acids and energy through walnut peptides and energy-providing peptides, promoting liver cell regeneration and constructing a regenerative system. Finally, it utilizes energy-releasing peptides to promote ATP synthesis, electrolyte balancers to maintain osmotic pressure, and alginate oligosaccharides to regulate intestinal absorption, collectively alleviating post-drinking fatigue and dehydration. This invention achieves a comprehensive effect of rapid hangover relief, liver protection, and damage prevention through a five-dimensional synergistic mechanism of "accelerated metabolism, liver cell protection, antioxidant effects, maintaining balance, and intestinal regulation." The scientifically formulated proportions of each component together construct a highly efficient and safe hangover relief and liver protection system. Detailed Implementation

[0017] To better understand the technical content of this invention, specific embodiments are provided below to further illustrate the invention.

[0018] Unless otherwise specified, the experimental methods used in the embodiments of this invention are all conventional methods.

[0019] Unless otherwise specified, all materials and reagents used in the embodiments of this invention are commercially available. Example 1

[0020] A complex peptide beverage for hangover relief and liver protection comprises the following ingredients by weight: 10 parts Hovenia dulcis oligopeptides, 5 parts walnut peptides, 18 parts antioxidant peptides, 8 parts energy-releasing co-peptides, 6 parts alcohol dehydrogenase activating peptides, 0.5 parts silymarin-serine chelate, 0.3 parts electrolyte balancer, 1 part alginate oligosaccharide, and 200 parts purified water. The antioxidant peptides are obtained by enzymatic hydrolysis of a mixture of Cordyceps militaris, Hippophae rhamnoides, and sunflower discs in a mass ratio of 14:8:3; the energy-releasing co-peptides are obtained by microbial fermentation of carnosine and arginine succinate peptides in a volume ratio of 1:0.2; the alcohol dehydrogenase activating peptides are obtained by fermentation of marine shellfish collagen; and the electrolyte balancer is sodium chloride.

[0021] The oligopeptides from Hovenia dulcis were prepared by the following method: Fresh, disease-free, and pollution-free Hovenia dulcis seeds were selected, dried, and then pulverized into fine powder. The powder was added to purified water at a ratio of g / mL of 1:1 and stirred until homogeneous. A compound enzyme was added and the mixture was hydrolyzed at 38℃ and pH 7.0 for 4 hours. After hydrolysis, the enzyme was inactivated at 80℃ for 15 minutes. The mixture was then centrifuged, and the supernatant was subjected to a three-stage gradient ultrafiltration. First, a 50 kDa membrane was used for ultrafiltration. The resulting filtrate ① was then ultrafiltered using a 10 kDa membrane, and the resulting filtrate ② was ultrafiltered using a 1 kDa membrane to obtain Hovenia dulcis oligopeptides with a molecular weight <1000 Da. The amount of compound enzyme added was 1.5% of the mass of the Hovenia dulcis powder. The compound enzyme consisted of Bacillus subtilis protease and alkaline protease in a mass ratio of 1:2, with an enzyme activity of 3000 IU / g.

[0022] The walnut peptide is prepared by the following method: Fresh, unspoiled walnuts are selected, the shells are removed to obtain walnut kernels, the kernels are crushed and passed through a 60-mesh sieve to obtain walnut meal powder. First, a hexane-ethanol solution with a volume ratio of 1.5:1 is used to defatt the walnut meal powder at 45°C with stirring for 35 minutes. Then, the defatted meal powder is filtered out, rinsed with purified water, and then soaked in a 0.1% sodium sulfite aqueous solution for 50 minutes. Water is then added, and the pH is adjusted to 7.0 with a dilute alkaline solution. Finally, the mixture is subjected to a process at 50°C. Ultrasonic extraction and centrifugation were performed to obtain supernatant a. The filter residue was subjected to the same ultrasonic extraction and centrifugation process to obtain supernatant b. Supernatants a and b were combined and concentrated to obtain a concentrate. Alkaline protease was added to the concentrate for the first enzymatic hydrolysis, followed by neutral protease for the second enzymatic hydrolysis. After the enzymatic hydrolysis, the enzyme was inactivated at 80℃ for 15 min to obtain the hydrolysate. The hydrolysate was precipitated, and the supernatant was collected. The supernatant was centrifuged to obtain supernatant c. Supernatant c was subjected to a two-stage gradient ultrafiltration. First, a 30 kDa membrane was used for ultrafiltration, and the resulting filtrate (③) was then ultrafiltered using a 3 kDa membrane to obtain walnut peptides. The enzyme dosage for the first enzymatic hydrolysis was 1.5% of the concentrate volume, the hydrolysis temperature was 30℃, the pH was 8.0, and the hydrolysis time was 6 h. The enzyme dosage for the second enzymatic hydrolysis was 0.5% of the concentrate volume, the hydrolysis temperature was 28℃, the pH was 6.0, and the hydrolysis time was 3.5 h.

[0023] The antioxidant peptides were prepared by the following method: Cordyceps militaris, Hippophae rhamnoides, and sunflower discs were mixed and washed in proportion, dried at 60℃ and pulverized into 50-mesh particles to obtain a mixed powder. Pure water was added to the mixed powder at a material-to-liquid ratio of 3:10 g / mL to obtain a mixed solution. The pH was adjusted to 7.0, and the solution was extracted at 90℃ for 70 min. After centrifugation, the supernatant was collected, and the pH was adjusted to 6.0. Bromelain, rhamnosidase, and aminopeptidase were added at a mass ratio of 1:0.5:0.1. The solution was stirred and hydrolyzed at 40℃ for 4 h. After hydrolysis, the enzymes were inactivated at 80℃ for 15 min to obtain the hydrolysate. The hydrolysate was centrifuged, and the supernatant was subjected to two-stage gradient ultrafiltration. First, a 10 kDa membrane was used for ultrafiltration, and the filtrate ④ was then ultrafiltered using a 2 kDa membrane. Subsequently, steam was introduced and the solution was conditioned at 90℃ for 5 min to obtain the antioxidant peptides. The total amount of bromelain, rhamnosidase, and aminopeptidase added was 1.0% of the mass of the mixed solution, and the enzyme activities of bromelain, rhamnosidase, and aminopeptidase were 3000 IU / g.

[0024] The energy-releasing co-peptide was prepared by the following method: Carnosine and arginine succinate peptide were mixed in a volume ratio, and microorganisms were added and fermented at 25°C for 72 hours. The supernatant was then collected by centrifugation to obtain the energy-releasing co-peptide. The microorganisms consisted of Bacillus and Lactobacillus in a mass ratio of 3.5:1.2, with a bacterial count of 2.0 × 10⁻⁶.8 CFU / g; the amino acid succinate peptide was prepared by the following method: L-arginine and succinic anhydride were mixed at a volume ratio of 1:1.0 and dissolved in phosphate buffer at pH 8.0. The mixture was stirred at 25°C for 8 hours. After the reaction, the pH was adjusted to 4.0 to precipitate the product. The precipitate was collected by centrifugation and washed with anhydrous ethanol at 4°C to obtain the arginine succinate monomer. The obtained arginine succinate monomer was dissolved in Tris-HCl buffer at pH 7.0-7.5 to prepare a 90 mM solution. Glutaminase and 0.1 M sodium chloride solution were added to obtain the reaction solution. The mixture was stirred at 35°C for 8 hours. The polymerization process was monitored by SDS-PAGE. When the molecular weight reached 1-2 kDa, the temperature was raised to 80°C and maintained for 15 minutes to terminate the reaction. The reaction solution was subjected to 2... Ultrafiltration was performed using a kDa membrane to obtain arginine succinate peptide; the amount of glutamine transaminase added was 2.00% of the mass of the reaction solution, and the enzyme activity of the glutamine transaminase was 400 IU / g.

[0025] The alcohol dehydrogenase activating peptide was prepared by the following method: Fresh marine shellfish were shelled, viscerated, and cleaned. Papain was used for enzymatic hydrolysis at pH 6.5 and 50℃ for 7 hours to obtain the hydrolysate. The hydrolysate was then inactivated at 85℃ for 15 minutes, followed by inoculation with *Lactobacillus plantarum* at pH 6.0, 35℃, and 75% humidity for 84 hours, with aeration and stirring every 12 hours. After fermentation, the mixture was centrifuged, and the supernatant was ultrafiltered at 3 kDa. The filtrate was then separated using a Sephadex G-25 gel chromatography column with 0.05 M phosphate buffer as the mobile phase. Fractions with retention times of 18-22 minutes were collected and further purified by reversed-phase high-performance liquid chromatography to obtain alcohol dehydrogenase activating peptides with molecular weights concentrated in the range of 500-1500 Da. The amount of papain added was 3% of the mass of the marine shellfish, with an enzyme activity of 4000 IU / g. The amount of Lactobacillus plantarum added was 4% of the mass of the hydrolysate, with a bacterial count of 5.0 × 10⁻⁶. 8 CFU / g.

[0026] The silybin-serine chelate was prepared by the following method: silybin and L-serine were dissolved in 50 mM Tris-HCl buffer at pH 7.5 at a mass ratio of 1:1.15. The mixture was stirred at 58°C in the dark for 8 h. The reaction solution was then filtered through a 0.2 μm filter and subsequently subjected to anion exchange chromatography on a DEAE Sepharose FF column. The fraction with a retention time of 15-18 min was collected. The collected fraction was placed in a 75% ethanol aqueous solution at 3°C ​​and allowed to stand for 15 h. The solid component was filtered and dried at 40°C for 8 h. The solid component was then pulverized and passed through a 100-mesh sieve to obtain the silybin-serine chelate. Example 2

[0027] A complex peptide beverage for hangover relief and liver protection comprises the following ingredients by weight: 14 parts Hovenia dulcis oligopeptides, 7.5 parts walnut peptides, 26.5 parts antioxidant peptides, 11.5 parts energy-releasing co-peptides, 7 parts alcohol dehydrogenase activating peptides, 1.2 parts silymarin-serine chelate, 0.5 parts electrolyte balancer, 2 parts alginate oligosaccharides, and 250 parts purified water. The antioxidant peptides are obtained by enzymatic hydrolysis of a mixture of Cordyceps militaris, Hippophae rhamnoides, and sunflower discs in a mass ratio of 20:9:4. The energy-releasing co-peptides are obtained by microbial fermentation of carnosine and arginine succinate peptides in a volume ratio of 2:0.2. The alcohol dehydrogenase activating peptides are obtained by fermentation of marine shellfish collagen. The electrolyte balancer consists of sodium chloride, sodium citrate, and zinc gluconate in a 1:1:1 ratio.

[0028] The oligopeptides from Hovenia dulcis were prepared by the following method: Fresh, disease-free, and pollution-free Hovenia dulcis seeds were selected, dried, and then pulverized into fine powder. The powder was added to purified water at a ratio of g / mL of 1:2 and stirred until homogeneous. A compound enzyme was added and hydrolyzed at 41.5℃ and pH 8.0 for 3 hours. After hydrolysis, the enzyme was inactivated at 85℃ for 12.5 minutes. The mixture was then centrifuged, and the supernatant was subjected to a three-stage gradient ultrafiltration. First, a 50 kDa membrane was used for ultrafiltration. The resulting filtrate ① was then ultrafiltered using a 10 kDa membrane, and the resulting filtrate ② was ultrafiltered using a 1 kDa membrane to obtain Hovenia dulcis oligopeptides with a molecular weight <1000 Da. The amount of compound enzyme added is 1.3% of the mass of the Hovenia dulcis powder. The compound enzyme is composed of Bacillus subtilis protease and alkaline protease in a mass ratio of 1:3.5. The enzyme activity of Bacillus subtilis protease and alkaline protease is 4000 IU / g.

[0029] The walnut peptide is prepared by the following method: Fresh, unspoiled walnuts are selected, the shells are removed to obtain walnut kernels, the kernels are crushed and passed through an 80-mesh sieve to obtain walnut meal powder. First, a hexane-ethanol solution with a volume ratio of 1.8:1 is used to defatt the walnut meal powder at 50°C with stirring for 30 minutes. Then, the defatted meal powder is filtered out, rinsed with purified water, and then soaked in a 0.2% sodium sulfite aqueous solution for 40 minutes. Water is then added, and the pH is adjusted to 8.0 with a dilute alkaline solution. Finally, the mixture is subjected to ultrafiltration at 55°C. Ultrasonic stirring extraction and centrifugation were performed to obtain supernatant a. The filter residue was subjected to the same ultrasonic stirring extraction and centrifugation process to obtain supernatant b. Supernatants a and b were combined and concentrated to obtain a concentrated solution. Alkaline protease was added to the concentrated solution for the first enzymatic hydrolysis, followed by neutral protease for the second enzymatic hydrolysis. After the enzymatic hydrolysis, the enzyme was inactivated at 85℃ for 12.5 min to obtain the hydrolysate. The hydrolysate was precipitated, and the supernatant was collected. The supernatant was centrifuged to obtain supernatant c. Supernatant c was subjected to two-stage gradient ultrafiltration. First, a 30 kDa membrane was used for ultrafiltration, and the filtrate ③ was then ultrafiltered using a 3 kDa membrane to obtain walnut peptides. The enzyme dosage for the first enzymatic hydrolysis was 1.5-2.0% of the concentrated solution volume, the hydrolysis temperature was 45℃, the pH was 8.5, and the hydrolysis time was 4.5 h. The enzyme dosage for the second enzymatic hydrolysis was 0.8% of the concentrated solution volume, the hydrolysis temperature was 30℃, the pH was 6.5, and the hydrolysis time was 3.0 h.

[0030] The antioxidant peptides were prepared by the following method: Cordyceps militaris, Hippophae rhamnoides, and sunflower discs were mixed and washed in proportion, dried at 70℃ and pulverized into 65-mesh particles to obtain a mixed powder. Pure water was added to the mixed powder at a ratio of g / mL of 4:10 to obtain a mixed solution. The pH was adjusted to 8.0, and the solution was extracted at 95℃ for 60 min. After centrifugation, the supernatant was collected, and the pH was adjusted to 6.5. Bromelain, rhamnosidase, and aminopeptidase were added at a mass ratio of 1:0.7:0.2. The solution was stirred and hydrolyzed at 50℃ for 3 h. After hydrolysis, the enzymes were inactivated at 85℃ for 12.5 min to obtain the hydrolysate. The hydrolysate was centrifuged, and the supernatant was subjected to two-stage gradient ultrafiltration. First, a 10 kDa membrane was used for ultrafiltration, and the filtrate ④ was then ultrafiltered using a 2 kDa membrane. Subsequently, steam was introduced and the solution was conditioned at 105℃ for 4 min to obtain the antioxidant peptides. The total amount of bromelain, rhamnosidase, and aminopeptidase added was 1.5% of the mass of the mixed solution, and the enzyme activities of bromelain, rhamnosidase, and aminopeptidase were 4000 IU / g.

[0031] The energy-releasing co-peptide was prepared by the following method: Carnosine and arginine succinate peptide were mixed in a volume ratio, and microorganisms were added and fermented at 31°C for 48 hours. The supernatant was then collected by centrifugation to obtain the energy-releasing co-peptide. The microorganisms consisted of Bacillus and Lactobacillus in a mass ratio of 5.9:2.1, with a cell viability of 5.0 × 10⁻⁶. 8 CFU / g; the amino acid succinate peptide was prepared by the following method: L-arginine and succinic anhydride were mixed at a volume ratio of 1:1.2 and dissolved in phosphate buffer at pH 8.5. The mixture was stirred at 27.5℃ for 7 h. After the reaction, the pH was adjusted to 4.5 to precipitate the product. The precipitate was collected by centrifugation and washed with anhydrous ethanol at 6℃ to obtain the arginine succinate monomer. The obtained arginine succinate monomer was dissolved in Tris-HCl buffer at pH 7.2 to prepare a 100 mM solution. Glutaminase and 0.1 M sodium chloride solution were added to obtain the reaction solution. The mixture was stirred at 37.5℃ for 7 h. The polymerization process was monitored by SDS-PAGE. When the molecular weight reached 1-2 kDa, the temperature was raised to 85℃ and maintained for 12.5 min to terminate the reaction. The reaction solution was subjected to 2 Ultrafiltration was performed using a kDa membrane to obtain arginine succinate peptide; the amount of glutamine transaminase added was 2.5% of the mass of the reaction solution, and the activity of the glutamine transaminase was 300 IU / g.

[0032] The alcohol dehydrogenase activating peptide was prepared by the following method: Fresh marine shellfish were shelled, viscerated, and cleaned. The shellfish were then enzymatically hydrolyzed with papain at pH 7.0 and 55℃ for 6 hours to obtain the hydrolysate. The hydrolysate was then inactivated at 87.5℃ for 12.5 minutes. Lactobacillus plantarum was then inoculated and fermented at pH 6.3, 37.5℃, and 80% humidity for 72 hours, with aeration and stirring every 12 hours. After fermentation, the mixture was centrifuged, and the supernatant was ultrafiltered at 3 kDa. The filtrate was then separated using a Sephadex G-25 gel chromatography column with 0.05 M phosphate buffer as the mobile phase. Fractions with retention times of 18-22 minutes were collected and further purified by reversed-phase high-performance liquid chromatography to obtain alcohol dehydrogenase activating peptides with molecular weights concentrated in the range of 500-1500 Da. The amount of papain added was 4% of the mass of the marine shellfish, with an enzyme activity of 4500 IU / g. The amount of *Lactobacillus plantarum* added was 5.5% of the mass of the hydrolysate, with a *Lactobacillus plantarum* viability count of 8.0 × 10⁻⁶. 8 CFU / g.

[0033] The silybin-serine chelate was prepared by the following method: silybin and L-serine were dissolved in 50 mM Tris-HCl buffer at pH 8.0 at a mass ratio of 1:1.3. The mixture was stirred at 60°C in the dark for 7 h. The reaction solution was then filtered through a 0.2 μm filter and subsequently subjected to anion exchange chromatography on a DEAE Sepharose FF column. The fraction with a retention time of 15-18 min was collected. The collected fraction was placed in an 80% ethanol aqueous solution at 4°C and allowed to stand for 12.5 h. The solid component was filtered and dried at 45°C for 7 h. The solid component was then pulverized and passed through a 120-mesh sieve to obtain the silybin-serine chelate. Example 3

[0034] A complex peptide beverage for hangover relief and liver protection comprises the following ingredients by weight: 18 parts Hovenia dulcis oligopeptides, 10 parts walnut peptides, 35 parts antioxidant peptides, 15 parts energy-releasing co-peptides, 8 parts alcohol dehydrogenase activating peptides, 2.0 parts silymarin-serine chelate, 0.8 parts electrolyte balancer, 3 parts alginate oligosaccharides, and 300 parts purified water. The antioxidant peptides are obtained by enzymatic hydrolysis of a mixture of Cordyceps militaris, Hippophae rhamnoides, and sunflower discs in a mass ratio of 25:10:5. The energy-releasing co-peptides are obtained by microbial fermentation of carnosine and arginine succinate peptides in a volume ratio of 3:0.2. The alcohol dehydrogenase activating peptides are obtained by fermentation of marine shellfish collagen. The electrolyte balancer is composed of potassium chloride, calcium lactate, and magnesium malate in a 1:1:1 ratio.

[0035] The oligopeptides from Hovenia dulcis were prepared by the following method: Fresh, disease-free, and pollution-free Hovenia dulcis seeds were selected, dried, and then pulverized into fine powder. The powder was added to purified water at a ratio of g / mL of 1:3 and stirred until homogeneous. A compound enzyme was added and the mixture was hydrolyzed at 45℃ and pH 9.0 for 2 hours. After hydrolysis, the enzyme was inactivated at 90℃ for 10 minutes. The mixture was then centrifuged, and the supernatant was subjected to a three-stage gradient ultrafiltration. First, a 50 kDa membrane was used for ultrafiltration. The resulting filtrate ① was then ultrafiltered using a 10 kDa membrane, and the resulting filtrate ② was ultrafiltered using a 1 kDa membrane to obtain Hovenia dulcis oligopeptides with a molecular weight <1000 Da. The amount of compound enzyme added was 3.0% of the mass of the Hovenia dulcis powder. The compound enzyme consisted of Bacillus subtilis protease and alkaline protease in a mass ratio of 1:5, with an enzyme activity of 5000 IU / g.

[0036] The walnut peptide is prepared by the following method: Fresh, unspoiled walnuts are selected, the shells are removed to obtain walnut kernels, the kernels are crushed and passed through a 100-mesh sieve to obtain walnut meal powder. First, a hexane-ethanol solution with a volume ratio of 2.0:1 is used to defatt the walnut meal powder at 55°C for 25 minutes with stirring. Then, the defatted meal powder is filtered out, rinsed with purified water, and then soaked in a 0.3% sodium sulfite aqueous solution for 30 minutes. Water is then added, and the pH is adjusted to 9.0 with a dilute alkaline solution. Finally, the mixture is subjected to a process at 60°C. Ultrasonic extraction and centrifugation were performed to obtain supernatant a. The filter residue was subjected to the same ultrasonic extraction and centrifugation process to obtain supernatant b. Supernatants a and b were combined and concentrated to obtain a concentrate. Alkaline protease was added to the concentrate for the first enzymatic hydrolysis, followed by neutral protease for the second enzymatic hydrolysis. After the enzymatic hydrolysis, the enzyme was inactivated at 90℃ for 10 min to obtain the hydrolysate. The hydrolysate was precipitated, and the supernatant was collected. The supernatant was centrifuged to obtain supernatant c. Supernatant c was subjected to a two-stage gradient ultrafiltration. First, a 30 kDa membrane was used for ultrafiltration, and the resulting filtrate (③) was then ultrafiltered using a 3 kDa membrane to obtain walnut peptides. The enzyme dosage for the first enzymatic hydrolysis was 2.0% of the concentrate volume, the hydrolysis temperature was 60℃, the pH was 9.0, and the hydrolysis time was 3 h. The enzyme dosage for the second enzymatic hydrolysis was 1.0% of the concentrate volume, the hydrolysis temperature was 38℃, the pH was 7.0, and the hydrolysis time was 2.5 h.

[0037] The antioxidant peptides were prepared by the following method: Cordyceps militaris, Hippophae rhamnoides, and sunflower discs were mixed and washed in proportion, dried at 80℃ and pulverized into 80-mesh particles to obtain a mixed powder. Pure water was added to the mixed powder at a material-to-liquid ratio of 5:10 g / mL to obtain a mixed solution. The pH was adjusted to 9.0, and the solution was extracted at 100℃ for 50 min. After centrifugation, the supernatant was collected, and the pH was adjusted to 7.0. Bromelain, rhamnosidase, and aminopeptidase were added at a mass ratio of 1:0.8:0.3. The solution was stirred at 60℃ for 2 h for enzymatic hydrolysis. After the enzymatic hydrolysis was completed, the enzymes were inactivated at 90℃ for 10 min to obtain the enzymatic hydrolysate. The enzymatic hydrolysate was centrifuged, and the supernatant was subjected to two-stage gradient ultrafiltration. First, a 10 kDa membrane was used for ultrafiltration, and the filtrate ④ was then ultrafiltered using a 2 kDa membrane. Subsequently, steam was introduced and the solution was conditioned at 120℃ for 3 min to obtain the antioxidant peptides. The total amount of bromelain, rhamnosidase, and aminopeptidase added was 2.0% of the mass of the mixed solution, and the enzyme activity of bromelain, rhamnosidase, and aminopeptidase was 5000 IU / g.

[0038] The energy-releasing co-peptide was prepared by the following method: Carnosine and arginine succinate peptide were mixed in a volume ratio, and microorganisms were added and fermented at 37°C for 24 hours. The supernatant was then collected by centrifugation to obtain the energy-releasing co-peptide. The microorganisms consisted of Bacillus and Lactobacillus in a mass ratio of 8.2:3.0, with a bacterial count of 9.0 × 10⁻⁶.8 CFU / g; the arginine succinate peptide was prepared by the following method: L-arginine and succinic anhydride were mixed at a volume ratio of 1:1.5 and dissolved in phosphate buffer at pH 9.0. The mixture was stirred at 30°C for 6 h. After the reaction, the pH was adjusted to 5.0 to precipitate the product. The precipitate was collected by centrifugation and washed with anhydrous ethanol at 8°C to obtain arginine succinate monomer. The obtained arginine succinate monomer was dissolved in Tris-HCl buffer at pH 7.5 to prepare a 110 mM solution. Glutamine transaminase and 0.1 M sodium chloride solution were added to obtain the reaction solution. The reaction was stirred at 40°C for 6 h. The polymerization process was monitored by SDS-PAGE. When the molecular weight reached 1-2 kDa, the temperature was raised to 90°C and maintained for 10 min to terminate the reaction. The reaction solution was ultrafiltered through a 2 kDa membrane to obtain arginine succinate peptide. The amount of glutamine transaminase added was 3.0% of the mass of the reaction solution, and the glutamine transaminase activity was 400. IU / g.

[0039] The alcohol dehydrogenase activating peptide was prepared by the following method: Fresh marine shellfish were shelled, viscerated, and cleaned. Papain was used for enzymatic hydrolysis at pH 7.5 and 60℃ for 5 hours to obtain the hydrolysate. The hydrolysate was then inactivated at 90℃ for 10 minutes. Lactobacillus plantarum was then inoculated and fermented at pH 6.5, 40℃, and 85% humidity for 60 hours, with aeration and stirring every 12 hours. After fermentation, the mixture was centrifuged, and the supernatant was ultrafiltered at 3 kDa. The filtrate was then separated using a Sephadex G-25 gel chromatography column with 0.05 M phosphate buffer as the mobile phase. Fractions with retention times of 18-22 minutes were collected and further purified by reversed-phase high-performance liquid chromatography to obtain alcohol dehydrogenase activating peptides with molecular weights concentrated in the range of 500-1500 Da. The amount of papain added was 5% of the mass of the marine shellfish, with an enzyme activity of 5000 IU / g. The amount of *Lactobacillus plantarum* added was 7% of the mass of the hydrolysate, with a *Lactobacillus plantarum* viability count of 1.0 × 10⁻⁶. 9 CFU / g.

[0040] The silybin-serine chelate was prepared by the following method: silybin and L-serine were dissolved in 50 mM Tris-HCl buffer at pH 8.5 at a mass ratio of 1:1.5. The mixture was stirred at 62°C in the dark for 6 h. The reaction solution was then filtered through a 0.3 μm filter and subsequently subjected to anion exchange chromatography on a DEAE Sepharose FF column. The fraction with a retention time of 15-18 min was collected. The collected fraction was placed in an 85% ethanol aqueous solution at 5°C and allowed to stand for 10 h. The solid component was filtered and dried at 50°C for 6 h. The solid component was then pulverized and passed through a 150-mesh sieve to obtain the silybin-serine chelate.

[0041] The hangover-relieving and liver-protecting compound peptide beverage described in Examples 1-3 is prepared by the following method, including the following steps: S1. Mix silymarin-serine chelate, electrolyte balancer, and alginate oligosaccharide according to the weight parts, add purified water preheated to 45℃, place in a stirrer and stir at 600rpm for 20min to obtain solution ①.

[0042] S2. Add Hovenia dulcis oligopeptide, walnut peptide, antioxidant peptide and energy release co-peptide to solution ① in sequence, stir at 80 rpm for 15 min, then slowly inject alcohol dehydrogenase activating peptide, and adjust the pH to 7.0 with 10% citric acid solution to obtain solution ②.

[0043] S3. Place solution ② into a homogenizer and homogenize it 3 times at 25MPa. Then sterilize it with UHT at 125℃ for 8 seconds. Then fill it to obtain the hangover relief and liver protection compound peptide drink. Example 4

[0044] Compared with Example 3, the difference in this embodiment is that the hangover-relieving and liver-protecting compound peptide beverage is prepared by the following method, including the following steps: S1. Mix silymarin-serine chelate, electrolyte balancer, and alginate oligosaccharide according to the weight parts, add purified water preheated to 40℃, place in a stirrer and stir at 800rpm for 25min to obtain solution ①.

[0045] S2. Add Hovenia dulcis oligopeptide, walnut peptide, antioxidant peptide and energy release co-peptide to solution ① in sequence, stir at 60 rpm for 20 min, then slowly inject alcohol dehydrogenase activating peptide, and adjust the pH to 6.8 with 8% citric acid solution to obtain solution ②.

[0046] S3. Place solution ② into a homogenizer and homogenize it three times at 20MPa. Then perform UHT sterilization at 120℃ for 10s. Then fill the bottle to obtain the hangover relief and liver protection compound peptide beverage. Example 5

[0047] Compared with Example 3, the difference in this embodiment is that the hangover-relieving and liver-protecting compound peptide beverage is prepared by the following method, including the following steps: S1. Mix silymarin-serine chelate, electrolyte balancer, and alginate oligosaccharide according to the weight parts, add purified water preheated to 50℃, place in a stirrer and stir at 800rpm for 1min to obtain solution ①.

[0048] S2. Add Hovenia dulcis oligopeptide, walnut peptide, antioxidant peptide and energy release co-peptide to solution ① in sequence, stir at 100 rpm for 10 min, then slowly inject alcohol dehydrogenase activating peptide, and adjust the pH to 7.2 with 6% sodium bicarbonate solution to obtain solution ②.

[0049] S3. Place solution ② into a homogenizer and homogenize twice at 30MPa, then sterilize with UHT at 130℃ for 5s, and then fill to obtain a hangover relief and liver protection compound peptide drink. Comparative Example 1

[0050] The difference between this comparative example and Example 3 is that the raw materials do not contain Hovenia dulcis oligopeptides. Comparative Example 2

[0051] The difference between this comparative example and Example 3 is that the raw materials do not contain energy-releasing co-peptides. Comparative Example 3

[0052] The difference between this comparative example and Example 3 is that the raw materials do not contain antioxidant peptides. Comparative Example 4

[0053] The difference between this comparative example and Example 3 is that the raw materials do not contain alcohol dehydrogenase activating peptide. Comparative Example 5

[0054] The difference between this comparative example and Example 3 is that the raw materials do not contain silymarin-serine chelate. Comparative Example 6

[0055] Compared with Example 3, the difference in this comparative example is that the hangover relief and liver protection compound peptide beverage includes the following ingredients by weight: 9 parts of Hovenia dulcis oligopeptide, 12 parts of walnut peptide, 18-35 parts of antioxidant peptide, 16 parts of energy release co-peptide, 4 parts of alcohol dehydrogenase activating peptide, 0.4 parts of silymarin-serine chelate, 1.0 part of electrolyte balancer, 5 parts of alginate oligosaccharide, and 310 parts of purified water.

[0056] Efficacy testing 1. Peptide content testing

[0057] Referring to GB 5009.5-2016 "National Food Safety Standard - Determination of Protein in Food", the Kjeldahl method was used in conjunction with HPLC correction factors to detect the peptide content of Hovenia dulcis oligopeptides, walnut peptides, antioxidant peptides, energy-releasing co-peptides, and alcohol dehydrogenase-activating peptides in the compound peptide beverages prepared in Examples 1-5. The values ​​were measured three times and averaged. The results are shown in Table 1.

[0058] Table 1

[0059] As can be seen from Table 1, the peptide content of the compound peptide beverages prepared in Examples 1-5, including jujube oligopeptide, walnut peptide, antioxidant peptide, energy release co-peptide, and alcohol dehydrogenase activating peptide, is all >90%.

[0060] 2. Test of hangover relief effect SPF-grade healthy male mice, weighing 20-25g, were selected and acclimatized for 7 days under conditions of 22℃, 50% humidity, and a 12h light-dark cycle, with free access to food and water. The mice were then randomly divided into an experimental group, a control group, and a blank control group, with 20 mice in each group. The experimental and control groups were administered 53% ABV liquor by gavage at 15mL / kg body weight. The experimental group was then administered the compound peptide drinks prepared in Examples 1-5 and Comparative Examples 1-6 by gavage at 15mL / kg body weight, while the control group received an equal volume of physiological saline. The blank control group received no gavage treatment. Blood ethanol and acetaldehyde concentrations were measured at 0.5h and 2h, respectively. At the target time point, 0.2-0.3 mL of whole blood was collected via the orbital venous plexus into heparin sodium anticoagulant tubes and stored on ice. 50 μL of blood sample was added to 200 μL of physiological saline and 5 μL of 2-propanol internal standard, respectively. After vortex centrifugation, the supernatant was collected and analyzed using headspace gas chromatography. The instrument's passing retention times were 3.2 min for ethanol and 2.8 min for acetaldehyde. The results are recorded as average values ​​in Table 2.

[0061] Table 2

[0062] As can be seen from Table 2, the ethanol and acetaldehyde contents of the experimental group were lower than those of the control group at 0.5h and 1h. In particular, Examples 1-5 of the present invention showed better results, indicating that the compound peptide beverage prepared by the present invention has a real and effective hangover relief effect.

[0063] 3. Liver protection effect test SPF-grade healthy male mice, weighing 20-25g, were selected and acclimatized for 7 days under conditions of 22℃, 50% humidity, and a 12h light-dark cycle, with free access to food and water. The mice were then randomly divided into experimental, control, and blank control groups, with 20 mice in each group. The experimental and control groups were administered 8 mL / kg of 30% ethanol aqueous solution by gavage. The experimental groups were then administered 10 mL / kg of the compound peptide drinks prepared in Examples 1-5 and Comparative Examples 1-6 by gavage, while the control group received an equal volume of physiological saline. The blank control group received no gavage treatment. The experimental and control groups were allowed free access to food and water and were administered the drinks by gavage for 14 consecutive days, while the blank control group was allowed free access to food and water for 14 days. On the last day after gavage, the mice were fasted for 24 hours. Blood was collected from the mice's eyes, and serum was separated. The activities of ALT and AST in the serum of each group of mice were measured using an alanine aminotransferase (ALT) assay kit and an aspartate aminotransferase (AST) assay kit, according to their respective instructions. After collecting blood from the eyeballs of mice in each group, they were dissected, and the livers were removed. 200 mg of liver tissue was added to ice-cold physiological saline for homogenization to prepare a 10% liver tissue homogenate. After vortex centrifugation, the supernatant was collected. The values ​​of glutathione (GSH), malondialdehyde (MDA), and superoxide dismutase (SOD) activity in the liver homogenate of each group of mice were measured according to their respective instructions using a reduced glutathione (GSH) assay kit, a malondialdehyde (MDA) assay kit, and a superoxide dismutase (SOD) activity assay kit. The test results are recorded as average values ​​in Table 3.

[0064] Table 3

[0065] As can be seen from Table 3, Examples 1-5 of the present invention have excellent liver-protecting effects compared with the control group, which shows that the compound peptide beverage prepared by the present invention has a real and effective liver-protecting effect.

[0066] This invention provides an alcohol dehydrogenase activating peptide that directly activates alcohol dehydrogenase, accelerating the conversion of ethanol to acetaldehyde. Hovenia dulcis oligopeptides inhibit intestinal absorption of ethanol, reducing its entry into the bloodstream, and upregulate the expression of alcohol dehydrogenase and acetaldehyde dehydrogenase genes, accelerating ethanol metabolism to acetic acid. The alcohol dehydrogenase activating peptide and Hovenia dulcis oligopeptides work synergistically to form a "dual-channel" ethanol metabolism acceleration effect. Hovenia dulcis oligopeptides also protect hepatocyte membranes through antioxidant effects, reducing alcohol-induced lipid peroxidation. Energy-releasing co-peptides provide nicotinamide adenine dinucleotide, increasing adenosine triphosphate (ATP) levels in the body, thereby improving the energy efficiency of alcohol breakdown. Walnut peptides contain various amino acids; arginine participates in the urea cycle, promoting ammonia metabolism, and glutamate can be converted into glutathione, enhancing the liver's antioxidant capacity. Furthermore, the polyunsaturated fatty acids in walnut peptides can help repair hepatocyte damage. Antioxidant peptides can scavenge reactive oxygen species (ROS) produced by alcohol metabolism, reducing lipid peroxidation and DNA damage. It can also inhibit the release of inflammatory factors and alleviate alcoholic liver damage. Silymarin-serine chelates, through the chelation of silymarin and serine, improve stability and absorption. They can also stabilize hepatocyte membranes and inhibit alcohol-induced apoptosis. Furthermore, they can promote hepatin synthesis and accelerate hepatocyte regeneration. In addition, serine provides metabolic substrates to aid liver detoxification. Alginate oligosaccharides adsorb acetaldehyde in the intestine, reducing enterohepatic toxicity and thus alleviating the burden on the liver. Electrolyte balancers replenish electrolytes lost due to alcohol metabolism, maintain neuromuscular function, and prevent dehydration and post-drinking muscle cramps.

[0067] This invention utilizes antioxidant peptides and alginate oligosaccharides to scavenge free radicals and toxins, reducing toxin entry into the liver and constructing a preventative system. It also employs silymarin-serine chelates to target and repair membrane structures, stabilizing organelle function and constructing a repair system. Furthermore, it provides amino acids and energy through walnut peptides and energy-providing peptides, promoting liver cell regeneration and constructing a regenerative system. Finally, it utilizes energy-releasing peptides to promote ATP synthesis, electrolyte balancers to maintain osmotic pressure, and alginate oligosaccharides to regulate intestinal absorption, collectively alleviating post-drinking fatigue and dehydration. This invention achieves a comprehensive effect of rapid hangover relief, liver protection, and damage prevention through a five-dimensional synergistic mechanism of "accelerated metabolism, liver cell protection, antioxidant effects, maintaining balance, and intestinal regulation." The scientifically formulated proportions of each component together construct a highly efficient and safe hangover relief and liver protection system.

[0068] 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 complex peptide beverage for relieving hangovers and protecting the liver, characterized in that, The raw materials include the following parts by weight: 10-18 parts of Hovenia dulcis oligopeptides, 5-10 parts of walnut peptides, 18-35 parts of antioxidant peptides, 8-15 parts of energy-releasing co-peptides, 6-8 parts of alcohol dehydrogenase activating peptides, 0.5-2.0 parts of silymarin-serine chelate, 0.3-0.8 parts of electrolyte balancer, 1-3 parts of alginate oligosaccharides, and 200-300 parts of purified water; the antioxidant peptides are obtained by enzymatic hydrolysis of Cordyceps militaris, Hippophae rhamnoides, and sunflower disc in a mass ratio of (14-25):(8-10):(3-5); the energy-releasing co-peptides are obtained by enzymatic hydrolysis of Cordyceps militaris, Hippophae rhamnoides, and sunflower disc in a volume ratio of 1-3: 0.2% carnosine and arginine succinate peptide were prepared by microbial fermentation; the alcohol dehydrogenase-activating peptide was prepared by fermentation of marine shellfish collagen.

2. The hangover-relieving and liver-protecting compound peptide beverage as described in claim 1, characterized in that, The oligopeptides from *Hovenia dulcis* seeds are prepared by the following method: Fresh, disease-free, and pollution-free *Hovenia dulcis* seeds are selected, dried, and then pulverized into fine powder. Pure water is added at a material-to-liquid ratio of 1:1-3 (g / mL) and stirred evenly. A compound enzyme is added and hydrolyzed at 38-45℃ and pH 7.0-9.0 for 2-4 hours. After hydrolysis, the enzyme is inactivated at 80-90℃ for 10-15 minutes. The mixture is then centrifuged, and the supernatant is subjected to a three-stage gradient ultrafiltration. First, a 50 kDa membrane is used for ultrafiltration. The resulting filtrate ① is then ultrafiltered using a 10 kDa membrane. The resulting filtrate ② is then ultrafiltered using a 10 kDa membrane. Ultrafiltration was performed using a kDa membrane to obtain Hovenia dulcis oligopeptides with a molecular weight <1000 Da; the amount of the complex enzyme added was 1.5-3.0% of the mass of the Hovenia dulcis powder, and the complex enzyme consisted of Bacillus subtilis protease and alkaline protease in a mass ratio of 1:2-5, with the enzyme activity of Bacillus subtilis protease and alkaline protease ≥3000 IU / g.

3. The hangover-relieving and liver-protecting compound peptide beverage as described in claim 1, characterized in that, The walnut peptide is prepared by the following method: Fresh, unspoiled walnuts are selected, the shells are removed to obtain walnut kernels, the kernels are crushed and passed through a 60-100 mesh sieve to obtain walnut meal powder. First, a hexane-ethanol solution with a volume ratio of 1.5-2.0:1 is used to defatt the walnut meal powder at 45-55℃ with stirring for 25-35 minutes. Then, the defatted meal powder is filtered out, rinsed with purified water, and then soaked in a 0.1-0.3% sodium sulfite aqueous solution for 30-50 minutes. Next, water is added, and the pH is adjusted to 7.0-9.0 with a dilute alkaline solution. Finally, the solution is soaked at 50- Ultrasonic extraction was performed at 60℃, followed by centrifugation to obtain supernatant a. The filter residue was subjected to the same ultrasonic extraction and centrifugation process to obtain supernatant b. Supernatants a and b were combined and concentrated to obtain a concentrate. Alkaline protease was added to the concentrate for the first enzymatic hydrolysis, followed by neutral protease for the second enzymatic hydrolysis. After the enzymatic hydrolysis, the enzymes were inactivated at 80-90℃ for 10-15 minutes to obtain the hydrolysate. The hydrolysate was precipitated, and the supernatant was collected. The supernatant was centrifuged to obtain supernatant c. Supernatant c was then subjected to a two-stage gradient ultrafiltration, first using 30... The solution was ultrafiltered using a 3 kDa membrane, and the resulting filtrate ③ was then ultrafiltered again using a 3 kDa membrane to obtain walnut peptides. The enzyme addition amount for the first enzymatic hydrolysis was 1.5-2.0% of the concentrated liquid volume, the hydrolysis temperature was 30-60℃, the pH was 8.0-9.0, and the hydrolysis time was 3-6 h. The enzyme addition amount for the second enzymatic hydrolysis was 0.5-1.0% of the concentrated liquid volume, the hydrolysis temperature was 28-38℃, the pH was 6.0-7.0, and the hydrolysis time was 2.5-3.5 h.

4. The hangover-relieving and liver-protecting compound peptide beverage as described in claim 1, characterized in that, The antioxidant peptides are prepared by the following method: Cordyceps militaris, Hippophae rhamnoides, and sunflower discs are mixed in proportion, washed, dried at 60-80℃ and pulverized into 50-80 mesh particles to obtain a mixed powder. Pure water is added to the mixed powder at a material-to-liquid ratio of 3-5:10 (g / mL) to obtain a mixed solution. The pH is adjusted to 7.0-9.0, and the solution is extracted at 90-100℃ for 50-70 minutes. After centrifugation, the supernatant is collected, and the pH is adjusted to 6.0-7.

0. Bromelain, rhamnosidase, and aminopeptidase are added at a mass ratio of 1:(0.5-0.8):(0.1-0.3). Enzymatic hydrolysis is carried out at 40-60℃ for 2-4 hours. After hydrolysis, the enzymes are inactivated at 80-90℃ for 10-15 minutes to obtain the hydrolysate. The hydrolysate is centrifuged, and the supernatant is subjected to two-stage gradient ultrafiltration. First, a 10 kDa membrane is used for ultrafiltration. The filtrate ④ is then subjected to two-stage gradient ultrafiltration. Ultrafiltration was performed using a kDa membrane, followed by conditioning with steam at 90-120℃ for 3-5 minutes to obtain antioxidant peptides.

5. The hangover-relieving and liver-protecting compound peptide beverage as described in claim 1, characterized in that, The energy-releasing co-peptide was prepared by the following method: carnosine and arginine succinate peptide were mixed in a volume ratio, added to microorganisms, and fermented at 25-37℃ for 24-72 hours. The supernatant was then collected by centrifugation to obtain the energy-releasing co-peptide. The microorganisms consist of Bacillus and Lactobacillus in a mass ratio of (3.5-8.2):(1.2-3.0), with a viable count ≥2.0×10⁻⁶. 8 CFU / g; The amino acid succinate peptide was prepared by the following method: L-arginine and succinic anhydride were mixed at a volume ratio of 1:1.0-1.5 and dissolved in phosphate buffer at pH 8.0-9.

0. The mixture was stirred at 25-30℃ for 6-8 h. After the reaction, the pH was adjusted to 4.0-5.0 to precipitate the product. The precipitate was collected by centrifugation and washed with anhydrous ethanol at 4-8℃ to obtain arginine succinate monomer. The obtained arginine succinate monomer was dissolved in Tris-HCl buffer at pH 7.0-7.5 to prepare a 90-110 mM solution. Glutaminase and 0.1 M sodium chloride solution were added to obtain the reaction solution. The mixture was stirred at 35-40℃ for 6-8 h. The polymerization process was monitored by SDS-PAGE. When the molecular weight reached 1-2 kDa, the temperature was raised to 80-90℃ and maintained for 10-15 min to terminate the reaction. The reaction solution was subjected to 2 Ultrafiltration was performed using a kDa membrane to obtain amino acid succinate peptides; the amount of glutamine transaminase added was 2.0-3.0% of the mass of the reaction solution, and the enzyme activity of the glutamine transaminase was 200-400 IU / g.

6. The hangover-relieving and liver-protecting compound peptide beverage as described in claim 1, characterized in that, The alcohol dehydrogenase-activating peptide was prepared by the following method: Fresh marine shellfish were shelled, viscerated, and cleaned. Papain was used for enzymatic hydrolysis at pH 6.5-7.5 and 50-60℃ for 5-7 hours to obtain the hydrolysate. The hydrolysate was then inactivated at 85-90℃ for 10-15 minutes. Lactobacillus plantarum was then inoculated and fermented at pH 6.0-6.5, 35-40℃, and 75-85% humidity for 60-84 hours, with aeration and stirring every 12 hours. After fermentation, the mixture was centrifuged, and the supernatant was ultrafiltered at 3 kDa. The filtrate was then separated using a Sephadex G-25 gel chromatography column with 0.05 M phosphate buffer as the mobile phase. Fractions with a retention time of 18-22 minutes were collected and further purified by reversed-phase high-performance liquid chromatography to obtain molecules with molecular weights concentrated in the range of 500-1500. Da's alcohol dehydrogenase-activating peptide; the amount of papain added is 3-5% of the mass of marine shellfish, the papain enzyme activity is ≥4000 IU / g, the amount of Lactobacillus plantarum added is 4-7% of the mass of the enzymatic hydrolysate, and the viable count of Lactobacillus plantarum is ≥5.0 × 10⁻⁶. 8 CFU / g.

7. The hangover-relieving and liver-protecting compound peptide beverage as described in claim 1, characterized in that, The silybin-serine chelate was prepared by the following method: silybin and L-serine were dissolved in 50 mM Tris-HCl buffer at a mass ratio of 1:1.1-1.5 at pH 7.5-8.5, and the mixture was stirred at 58-62°C in the dark for 6-8 h. The reaction solution was then filtered through a 0.2-0.3 μm filter and subsequently subjected to anion exchange chromatography using a DEAE Sepharose FF column. The fraction with a retention time of 15-18 min was collected. The collected fraction was placed in a 75-85% ethanol aqueous solution at 3-5°C and allowed to stand for 10-15 h. The solid component was filtered and dried at 40-50°C for 6-8 h. The solid component was then pulverized and passed through a 100-150 mesh sieve to obtain the silybin-serine chelate.

8. The hangover-relieving and liver-protecting compound peptide beverage as described in claim 1, characterized in that, The electrolyte balancer is any one or a combination of several of sodium chloride, potassium chloride, calcium lactate, sodium citrate, magnesium malate, and zinc gluconate.

9. A method for preparing a hangover-relieving and liver-protecting compound peptide beverage as described in any one of claims 1-8, characterized in that, Includes the following steps: S1. Mix silymarin-serine chelate, electrolyte balancer, and alginate oligosaccharide according to the weight parts, add purified water preheated to 40-50℃, place in a stirrer and stir at 400-800 rpm for 15-25 min to obtain solution ①. S2. Add the Hovenia dulcis oligopeptide, walnut peptide, antioxidant peptide and energy release co-peptide to solution ① in sequence, stir at 60-100 rpm for 10-20 min, then slowly inject the alcohol dehydrogenase activating peptide, and adjust the pH to 6.8-7.2 with 8-12% citric acid solution or 4-6% sodium bicarbonate solution to obtain solution ②. S3. Place solution ② into a homogenizer and homogenize it 2-3 times at 20-30MPa. Then perform UHT sterilization at 120-130℃ for 5-10 seconds. After that, fill the bottle to obtain the hangover relief and liver protection compound peptide beverage.

10. A hangover-relieving and liver-protecting compound peptide beverage as described in any one of claims 1-8, characterized in that, The compound peptide beverage is used as a hangover relief and liver protection health product or medicine.

Images