Probiotic anti-alcohol capsule as well as preparation method and application thereof

Abstract

The invention discloses a probiotic hangover-alleviating capsule as well as a preparation method and application thereof, and belongs to the technical field of health-care functional foods. The preparation method comprises the following steps: (1) stirring and mixing a probiotic solution and a zinc salt solution, then adding an EGCG solution, and continuously stirring and mixing; (2) adding the solution obtained in the step (1) into a sodium alginate solution, stirring and mixing, then adding into a calcium lactate solution, vibrating and hardening, and filtering to obtain probiotic gel; and (3) freeze-drying the probiotic gel, and taking gelatin as a coating to prepare the probiotic hangover-alleviating capsule. The probiotic capsule designed by the invention has excellent targeting property, and is beneficial to successful colonization of probiotics in small intestines. The specially designed double-layer shell structure not only can slow down the absorption of the gastrointestinal tract to alcohol and protect the gastric mucosa, but also can effectively relieve intestinal injury and prevent dysbacteriosis.

Description

Technical Field

[0001] The invention belongs to the technical field of health functional foods, and particularly relates to a probiotic alcohol-resolving capsule and a preparation method and application thereof. Background Art

[0002] Drinking is a common social behavior worldwide. Whether in social occasions, celebrations or daily life, alcoholic beverages are widely consumed. However, excessive drinking or long-term alcoholism can cause serious harm to human health. Studies have shown that alcohol mainly enters the blood through the stomach (absorption of about 10% to 20%) and the small intestine (absorption of about 80% to 90%), and is then metabolized in the liver by alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH). In this process, the byproducts produced (such as acetaldehyde, free radicals and lactic acid) often cause a variety of physiological problems, such as headaches, nausea, vomiting, dizziness, and even more serious liver damage, oxidative stress and inflammatory responses. Therefore, excessive drinking can not only cause acute alcohol poisoning, but may also lead to chronic diseases such as fatty liver, cirrhosis, cardiovascular disease and nervous system damage.

[0003] Alcohol remedies are a type of medicine or health product that aims to relieve or eliminate the discomfort symptoms after drinking, and reduce the damage of alcohol to the body by promoting alcohol metabolism, protecting the liver and other mechanisms. Its main functions include: enhancing the activity of enzymes related to alcohol metabolism in the body, accelerating the decomposition of ethanol and acetaldehyde, thereby shortening the residence time of alcohol in the body; protecting the liver from damage caused by acetaldehyde accumulation through antioxidant and anti-inflammatory effects or promoting liver cell repair; at the same time, by replenishing nutrients consumed by drinking (such as vitamins and minerals) or regulating the water balance in the body, it can relieve discomfort symptoms such as headaches and nausea.

[0004] At present, most of the hangover remedies sold on the market are "health care drugs". Their health care effects rely on flavonoids, anthocyanins and phenolic compounds contained in Chinese medicinal materials, tea and fruit extracts. These compounds can promote gastrointestinal motility and reduce alcohol damage to the digestive tract mucosa and liver, thereby reducing the harm of drinking to the human body, but they cannot essentially eliminate alcohol or reverse the alcohol metabolism process in the human body.

[0005] Alcoholic probiotics are a type of probiotics that are highly tolerant to alcohol. They can survive and reproduce in a certain concentration of alcohol, while continuing to regulate intestinal flora, promote digestion, and enhance immune function. In addition, some functional strains rely on their own unique enzyme system to assist the host in breaking down alcohol in the body. However, excessive gastric acid secretion caused by drinking will increase the difficulty of successful colonization of alcoholic probiotics in the intestine to a certain extent. Therefore, how to improve the survival rate of probiotics in the gastric acid environment, prolong their residence time in the small intestine, and enhance their colonization rate in the small intestine is an urgent problem to be solved. Summary of the invention

[0006] In view of the above-mentioned deficiencies in the prior art, the present invention provides a probiotic hangover-relieving capsule and its preparation method and use. The present invention adopts a double-layer coating technology to transform the probiotics. The inner layer is composed of zinc-based metal-epigallocatechin-gallate (Zn-EGCG) which is sensitive to alkali but not sensitive to acid, and the outer layer is a hydrogel with a vesicular, loose and porous structure formed by sodium alginate (SA) and calcium lactate. It can ensure that the probiotics can be effectively protected when passing through the stomach and can be released after successfully reaching the small intestine, so as to give full play to their adhesion and colonization effects.

[0007] To achieve the above purpose, the technical solution adopted by the present invention to solve the technical problem is: The purpose of the present invention is to provide a method for preparing a probiotic alcohol-relieving capsule, comprising the following steps: (1) Stir the probiotic solution and the zinc salt solution, then add the EGCG solution and continue stirring and mixing; (2) adding the sodium alginate solution to the solution obtained in step (1), stirring and mixing, and then adding the calcium lactate solution, shaking and hardening, filtering, and obtaining a probiotic gel; (3) The probiotic gel is freeze-dried and coated with gelatin to prepare probiotic hangover-relieving capsules.

[0008] Furthermore, in step (1), the probiotics are dissolved in a freeze-dried protective agent solution to obtain a probiotic solution.

[0009] Furthermore, the probiotics are probiotics that have the effects of sobering up and protecting the liver.

[0010] Furthermore, the probiotics are Lactobacillus rhamnosus (CGMCC No. 27226, which has been disclosed in the invention patent with application number CN202311255212.X), Pediococcus acidilactici (CGMCC No. 20087, which has been disclosed in the invention patent with application number CN202410386836.3), Lactobacillus plantarum (CGMCC No. 25371, which has been disclosed in the invention patent with application number CN202410386836.3), and Pediococcus pentosaceus (CGMCC No. 27674, which has been disclosed in the invention patent with application number CN202410797299.1).

[0011] Furthermore, the lyophilization protectant solution is a sucrose solution with a concentration of 1.5-5%.

[0012] Furthermore, the concentration of the sucrose solution is 5%.

[0013] Furthermore, in step (1), the volume ratio of the probiotic solution to the zinc salt solution is 10-12:1.

[0014] Furthermore, the volume ratio of the probiotic solution to the zinc salt solution is 12:1.

[0015] Furthermore, the zinc salt solution is a zinc sulfate solution with a concentration of 1-1.5 mg / mL.

[0016] Furthermore, the concentration of the zinc salt solution is 1.25 mg / mL.

[0017] Furthermore, in step (1), the volume ratio of the EGCG solution to the zinc salt solution is 1-2:1.

[0018] Furthermore, the concentration of the EGCG solution was 5 mg / mL, and the volume ratio of the EGCG solution to the zinc salt solution was 2:1.

[0019] Furthermore, in step (2), the volume ratio of the solution obtained in step (1) to the sodium alginate solution is 0.5-1:1.

[0020] Furthermore, the volume ratio of the solution obtained in step (1) to the sodium alginate solution is 1:1.

[0021] Furthermore, the amount of calcium lactate solution used in step (2) is 3 to 4 times the volume of the mixed solution containing sodium alginate.

[0022] Another object of the present invention is to provide a probiotic alcohol-relieving capsule, which is prepared by the above method.

[0023] Another object of the present invention is to provide a probiotic hangover-relieving capsule, which comprises probiotics, an inner coating layer, an outer coating layer and a coating layer from the inside to the outside; The inner coating is composed of zinc-based metal-epigallocatechin-gallate, which is sensitive to base but insensitive to acid; The outer coating layer is composed of calcium alginate hydrogel formed by sodium alginate and calcium lactate; The coating layer is gelatin.

[0024] Another object of the present invention is to provide the use of the above-mentioned probiotic alcohol-relieving capsule in the preparation of a preparation having the effects of protecting the gastrointestinal tract, protecting the liver and relieving alcohol.

[0025] Beneficial effects of the present invention: The present invention has found through research that the hydroxyl groups (-OH) at positions 5 and 7 of the A ring of EGCG, the hydroxyl groups (-OH) at positions 3' and 4' of the B ring, the hydroxyl group (-OH) at position 3 of the C ring, and the hydroxyl groups (-OH) at positions 4 and 6 of the gallic acid group can interact with various materials, thereby enhancing the stability of the materials. Therefore, in the process of forming the metal-EGCG complex, probiotics can be introduced to form a probiotic coating film by complexation, thereby increasing the probability of the probiotics successfully passing through the stomach.

[0026] In the presence of calcium ions, alginate can form a gel with a three-dimensional network structure through ion cross-linking reaction. It has good biocompatibility and mechanical strength, can be degraded and absorbed by the human body, and has no toxic side effects (its degradation products, such as mannose and gulose, are harmless to the human body). Therefore, in the preparation process of calcium alginate hydrogel, by introducing metal-EGCG-coated probiotics, the probiotics can be further wrapped by cross-linking to form a second layer of coating. Ensure that the probiotics can be effectively protected when passing through the stomach and released after successfully reaching the small intestine, so as to give full play to their adhesion and colonization effects.

[0027] When probiotics are encapsulated with a metal-EGCG shell and calcium alginate hydrogel, the survival rate of probiotics in a gastric acid environment can be significantly improved, and their residence time in the intestine can be prolonged, thereby increasing the probability of probiotics colonizing in the small intestine. At the same time, the inner layer of metal-EGCG not only helps to stabilize the intestinal flora, but also has the effect of reducing intestinal inflammation. In addition, the outermost layer of hydrogel composed of sodium alginate has a loose and porous structure, which can absorb alcohol entering the gastrointestinal tract like a "buffer cotton", enhance gastrointestinal motility, and promote the excretion of alcohol.

[0028] The probiotic capsule designed by the present invention has excellent targeting, which is conducive to the successful colonization of probiotics in the small intestine. The specially designed double-layer shell structure can not only slow down the absorption of alcohol by the gastrointestinal tract and protect the gastric mucosa, but also effectively reduce intestinal damage and prevent dysbacteriosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 The blood alcohol concentration test results; Figure 2 The results of the survival rate test of bacteria in the acidic environment simulating gastric fluid. DETAILED DESCRIPTION

[0030] The specific implementation modes of the present invention are described below so that those skilled in the art can understand the present invention. However, it should be clear that the present invention is not limited to the scope of the specific implementation modes. For those of ordinary skill in the art, as long as various changes are within the spirit and scope of the present invention as defined and determined by the attached claims, these changes are obvious, and all inventions and creations utilizing the concept of the present invention are protected.

[0031] Example 1 A probiotic alcohol-relieving capsule, the preparation method of which is as follows: (1) Dissolve Lactobacillus rhamnosus (CGMCC No. 27226) in 5% sucrose solution to prepare mixed solution A; (2) Mix the mixture A and ZnSO4 (1.25 mg / mL) solutions were mixed at a volume ratio of 12:1 and vortexed for 10 s to prepare mixed solution B; (3) Mixture B and EGCG (5 mg / mL) solution were mixed at a volume ratio of 13:2 and vortexed for 10 s to prepare mixture C; (4) Add mixed solution C to 1% sodium alginate solution at a volume ratio of 1:1 and vortex for 10 seconds to prepare mixed solution D; (5) Add the mixed solution D dropwise to 4 times the volume of 1% calcium lactate solution, shake and harden for 1 hour, then wash and filter with water to prepare a probiotic gel; (6) The probiotic gel is freeze-dried and filled into a gelatin shell to prepare a probiotic hangover-relieving capsule.

[0032] Example 2 A probiotic alcohol-relieving capsule, the preparation method of which is as follows: (1) Dissolve Pediococcus acidilactici (CGMCC No. 20087) in 5% sucrose solution to prepare mixed solution A; (2) Mix the mixture A and ZnSO 4 (1.5 mg / mL) solutions were mixed at a volume ratio of 10:1 and vortexed for 10 s to prepare mixed solution B; (3) Mixed solution B and EGCG (5 mg / mL) solution were mixed at a volume ratio of 11:2 and vortexed for 10 s to prepare mixed solution C; (4) Add mixed solution C to 1% sodium alginate solution at a volume ratio of 0.5:1 and vortex for 10 seconds to prepare mixed solution D; (5) Add the mixed solution D dropwise to 4 times the volume of 1% calcium lactate solution, shake and harden for 1 hour, then wash and filter with water to prepare a probiotic gel; (6) The probiotic gel is freeze-dried and filled into a gelatin shell to prepare a probiotic hangover-relieving capsule.

[0033] Example 3 A probiotic alcohol-relieving capsule, the preparation method of which is as follows: (1) Dissolve Lactobacillus plantarum (CGMCC No. 25371) in 5% sucrose solution to prepare mixed solution A; (2) Mix the mixture A and ZnSO 4 (1.3 mg / mL) solutions were mixed at a volume ratio of 11:1 and vortexed for 10 s to prepare mixed solution B; (3) Mixed solution B and EGCG (5 mg / mL) solution were mixed at a volume ratio of 12:2 and vortexed for 10 s to prepare mixed solution C; (4) Add the mixed solution C to the 1% sodium alginate solution at a volume ratio of 0.8:1 and vortex for 10 s to prepare a mixed solution D; (5) Drop the mixed solution D into 4 times the volume of 1% calcium lactate solution, shake and harden for 1 hour, then wash with distilled water, filter, and prepare the probiotic gel; (6) Freeze-dry the probiotic gel and load it into a gelatin shell to prepare the probiotic anti-alcohol capsule.

[0034] Test Example I. Verification experiment on C57BL / 6 mice, the specific process is as follows: 1. Adaptive feeding of mice Mice were adaptively fed for 7 days before the experiment, with the temperature controlled at 22 - 25 °C, humidity 40% - 60%, and free access to water and food.

[0035] 2. Alcohol gavage: (1) Mice were fasted for 12 hours and had free access to water.

[0036] (2) 2 h before the start of the experiment, different drugs were administered by gavage according to the grouping.

[0037] Mice in experimental group 1 were administered the probiotic anti-alcohol capsule prepared in Example 1 of the present invention and given a small amount of normal saline; Mice in experimental group 2 were administered the Alcoburner anti-alcohol drug and given a small amount of normal saline; Mice in control group 1 were administered the same dose of dietary fiber and given a small amount of normal saline; Mice in control group 2 were only given a small amount of normal saline.

[0038] (3) According to the grouping, different doses of alcohol were administered by gavage to the mice.

[0039] Control group: Given the same volume of normal saline; Low-dose group: 5% (v / v) alcohol dose; High-dose group: 50% (v / v) alcohol dose; The gavage volume was 0.1 mL / 10 g body weight. After gavage, the mice were put back into the cage for observation 3. Determination of blood alcohol concentration At 0, 0.5, 1, 2, 4, 6, and 8 hours after gavage, the mice were anesthetized and blood was taken to detect the blood alcohol concentration of each treatment. The results are shown in Figure 1 ,, Figure 1 In figure A of, the detection results of the low-dose group are shown, and in figure B are the detection results of the high-dose group.

[0040] As Figure 1 shown, the anti-alcohol capsule prepared by the present invention can rapidly reduce the alcohol content in the blood and has excellent anti-alcohol effects.

[0041] 2. Bacteria plate verification experiment, the specific process is as follows 1. Acquisition of strains Under sterile conditions, the culture solution of Lactobacillus rhamnosus was centrifuged at 4000 g. After centrifugation, the supernatant was carefully poured off, the bacterial precipitate was collected, and it was resuspended in an appropriate amount of culture medium, mixed and set aside.

[0042] 2. Simulated gastric juice treatment The experiment is divided into 6 groups, and the specific settings are as follows: Blank group: Use 9 mL of culture medium with pH = 6.5-7, add 1 mL of resuspended bacterial solution, and then incubate at 30°C for 1 hour.

[0043] CK group (control group): Use 9 mL of culture medium with pH = 1.5~2, add 1 mL of resuspended bacterial solution, and then incubate at 30℃ for 1 hour.

[0044] EGCG group: Use 9 mL of culture medium with pH = 1.5~2, add 1 mL of EGCG@ resuspended bacterial solution, and then incubate at 30℃ for 1 hour.

[0045] EGCG-Zn group (EGCG-Zn cross-linking group): 9 mL of culture medium with pH = 1.5~2 was used, followed by adding 1 mL of EGCG-Zn@ resuspended bacterial solution and incubating at 30°C for 1 hour.

[0046] SA group (sodium alginate group): 9 mL of culture medium with pH = 1.5~2 was used, and 1 mL of SA@ resuspended bacterial solution was added, followed by incubation at 30°C for 1 hour.

[0047] SA@EGCG-Zn group (the probiotic alcohol-detoxifying capsule product prepared in Example 1): 9 mL of culture medium with pH = 1.5~2 was used, and 1 mL of SA@EGCG-Zn@ resuspended bacterial solution was added, followed by incubation at 30°C for 1 hour.

[0048] 3. Bacteria survival rate test Under sterile conditions, after 30 minutes and 60 minutes of treatment, 1 mL of each treatment group solution was added to 99 mL of culture medium to neutralize the acidic environment of simulated gastric juice and terminate the gastric juice stress treatment. Subsequently, the treated bacterial solution was continuously diluted 10 times, 100 times, and 1000 times.

[0049] Take 1 mL of each diluted bacterial solution and evenly spread it on the surface of the solid culture medium. Place the culture dish in an incubator at 30°C for 24 hours, then observe and record the growth of bacteria in each treatment group to evaluate the survival rate of bacteria under different conditions. The results are shown in Figure 2 The calculation formula is: survival rate = experimental group / blank group × 100%.

[0050] according to Figure 2 The test results show that in the acidic environment of simulated gastric juice, the survival rate of the bacteria in the probiotic capsules prepared by the present invention is significantly better than that of other control groups and experimental groups, indicating that the technical solution of the present invention can ensure that the probiotics can be effectively protected when passing through the stomach, so that they can be released smoothly after reaching the small intestine.

[0051] Finally, it should be noted that the above specific implementation methods are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to examples, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the spirit and scope of the technical solution of the present invention, which should be included in the scope of the claims of the present invention.

Claims

1. A method for preparing a probiotic alcohol-relieving capsule, characterized in that: The following steps are involved: (1) Stir and mix the probiotic solution and the zinc salt solution, then add the EGCG solution and continue stirring and mixing; (2) adding the solution obtained in step (1) to the sodium alginate solution, stirring and mixing, and then adding to the calcium lactate solution, shaking and hardening, filtering, and obtaining a probiotic gel; (3) The probiotic gel is freeze-dried and coated with gelatin to prepare probiotic hangover-relieving capsules.

2. The preparation method according to claim 1, characterized in that: In step (1), the probiotics are dissolved in a freeze-dried protective agent solution to obtain a probiotic solution.

3. The preparation method according to claim 2, characterized in that: The protective agent solution is a sucrose solution with a concentration of 1.5-5%.

4. The preparation method according to claim 1, characterized in that: In step (1), the volume ratio of the probiotic solution to the zinc salt solution is 10-12:

1.

5. The preparation method according to claim 1 or 4, characterized in that: The zinc salt solution is a zinc sulfate solution with a concentration of 1-1.5 mg / mL.

6. The preparation method according to claim 1, characterized in that: In step (1), the volume ratio of the EGCG solution to the zinc salt solution is 1-2:

1.

7. The preparation method according to claim 1, characterized in that: In step (2), the volume ratio of the solution obtained in step (1) to the sodium alginate solution is 0.5-1:1; and the amount of calcium lactate solution used is 3-4 times the volume of the mixed solution containing sodium alginate.

8. A probiotic alcohol-relief capsule, characterized in that: The method is prepared by any one of claims 1 to 7.

9. A probiotic alcohol-relieving capsule, characterized in that: From the inside to the outside, it includes probiotics, inner coating layer, outer coating layer and coating layer; The inner coating layer is composed of zinc-based metal-epigallocatechin-gallate which is sensitive to alkali but insensitive to acid; The outer coating layer is composed of calcium alginate hydrogel formed by sodium alginate and calcium lactate; The coating layer is gelatin.

10. Use of the probiotic alcohol-detoxifying capsule according to claim 8 or 9 in the preparation of a preparation having gastrointestinal protection, liver protection and alcohol-detoxifying effects.

Images