A fermented beverage and its preparation method

Abstract

This invention relates to a fermented beverage and its preparation method, belonging to the field of fermented beverage technology, with IPC classification number A23L2 / 38. The invention uses wolfberry, cassia seed, burdock root, mulberry bark, and codonopsis as raw materials. These are boiled in water, and then inoculated with specific strains of *Candida krusei* CICC 31807, *Saccharomyces cerevisiae* CICC 31105, and *Bifidobacterium animalis* CICC 6165 in a ratio of (2-5):1:(2.5-6) ​​for fermentation. After post-processing, a concentrated material is obtained. This concentrated material is then combined with *Phyllanthus emblica* extract, mulberry extract, sweetener, acidity regulator, excipients, and water to prepare the fermented beverage. The beverage prepared by this invention has good stability, maintaining good stability even after exposure to high or low temperatures; it has a rich aroma, no bitterness, and a smooth, pleasant taste, providing a refreshing and delicate experience, and also has a certain liver-protective effect.

Description

Technical Field

[0001] This invention relates to a fermented beverage and its preparation method, belonging to the field of fermented beverage technology, with IPC classification number A23L2 / 38. Background Technology

[0002] Fermented beverages are beverages with low alcohol content that are formulated through microbial fermentation. In fermented beverages, the biochemical fermentation process of microorganisms significantly increases the effective nutritional components of the fermented raw materials, enhancing the variety and abundance of active ingredients such as amino acids, peptides, free minerals, and organic matter, thus compensating for the nutritional deficiencies of beverages obtained through conventional juicing processes.

[0003] Meanwhile, during the fermentation process, microorganisms cause a series of complex biochemical changes in raw materials that are both food and medicine, generating many active substances that have good health benefits for the human body. For example, they can regulate the pH value in the gastrointestinal tract, regulate gastrointestinal function, help digestion and absorption, cleanse the intestines, and inhibit the growth of harmful bacteria. The generated amino acids, peptides, flavonoids, and other organic substances can maintain liver function and intestinal health, lower blood pressure, break down cholesterol, and prevent arteriosclerosis and cardiovascular diseases. They can also eliminate fatigue and muscle pain, increase appetite, promote digestion, protect the skin, promote calcium absorption, and have a detoxification effect.

[0004] In addition, fermented beverages undergo a series of beneficial changes in flavor, taste, and color under the action of microorganisms. This can reduce the fishy and bitter taste of raw materials, increase the color of beverages, make the taste of beverages smooth and delicate, and enhance the flavor.

[0005] With the changing pace of life and dietary habits, staying up late, drinking alcohol, and irregular eating habits all place a significant burden on the liver, leading to liver damage. While medication can effectively treat and prevent liver damage, it is inconvenient to use and has certain side effects. Therefore, providing a daily beverage that also offers nutrition and liver protection is a topic that requires further research. Summary of the Invention

[0006] To address the aforementioned technical problems, a fermented beverage and its preparation method are proposed. The technology belongs to the field of fermented beverage technology and is classified under IPC number A23L2 / 38.

[0007] This invention provides a method for preparing a fermented beverage, comprising the following steps:

[0008] Step 1: Wash and dry the wolfberries, cassia seeds, burdock root, mulberry bark, and codonopsis root, then crush them to obtain crushed materials;

[0009] Step 2: Add water to the crushed material and boil it, then cool it to obtain a mixture;

[0010] Step 3: Mix the mixture with sugar to obtain a treatment solution, inoculate with fermentation bacteria, ferment, sterilize, and obtain fermented product;

[0011] Step 4: Filter and concentrate the fermented material to obtain concentrated material;

[0012] Step 5: Add amla extract, mulberry extract, sweetener, acidity regulator, excipients and water to the concentrated material, stir well, sterilize, and age to obtain the beverage intermediate;

[0013] Step 6: Fill and package the beverage intermediate to obtain the fermented beverage.

[0014] Preferably, in step 1, the mass fractions of each raw material are: 0.1-1 parts wolfberry, 0.15-1.2 parts cassia seed, 0.13-1.5 parts burdock root, 0.1-0.8 parts mulberry bark, and 0.1-0.85 parts codonopsis.

[0015] Preferably, in step 1, the mass ratio of cassia seed to burdock root is 1:0.7-1.2, 1:0.75-1.2, 1:0.8-1.15, 1:0.95-1.1, etc.

[0016] Preferably, in step 1, the particles are crushed to a size of less than 1 cm.

[0017] Preferably, in step 2, the mass ratio of crushed material to water is 1:10-100, 1:10-90, 1:10-50, 1:12-40, etc.

[0018] Preferably, in step 2, the temperature is cooled to 20-40℃, 22-38℃, 22-35℃, 25-30℃, etc.

[0019] Preferably, in step 3, the mass of sugar is 1-4%, 1.2-4%, 1.5-3.8%, or 2-3.5% of the mixture.

[0020] Preferably, in step 3, the sugar includes at least one of glucose, sucrose, and fructose.

[0021] Preferably, in step 3, the fermentation bacteria include a mixture of Candida cruzie CICC 31807, Saccharomyces cerevisiae CICC 31105 and Bifidobacterium animalis CICC 6165.

[0022] Preferably, in step 3, the inoculum size of the fermentation bacteria in each mL of treatment solution is 1 × 10⁻⁶. 9 -9×10 9 CFU, 1.5×10 9 -9×10 9 CFU, 2×10 9 -8.5×10 9CFU, 3×10 9 -8×10 9 CFU, etc.

[0023] Preferably, in step 3, the ratio of the number of Candida crus-galli CICC 31807, Saccharomyces cerevisiae CICC 31105 and Bifidobacterium animalis CICC 6165 in the fermentation bacteria is (2-5):1:(2.5-6), (2.5-5):1:(3-6), (3-4.5):1:(3.2-5.5), (3.5-4.5):1:(4-5.5), etc.

[0024] Preferably, in step 3, the fermentation temperature is 30-45℃, 30-43℃, 30-40℃, 32-38℃, 35℃, etc.

[0025] Preferably, in step 3, the fermentation time is 7-10 days, 7.5-10 days, 7.5-9.5 days, or 8-9 days.

[0026] Preferably, in step 4, the concentration is performed using vacuum concentration.

[0027] Preferably, in step 4, the material is concentrated to a mass of 8-20%, 9-18%, 10-17%, or 11-15% of the fermentation material.

[0028] Preferably, in step 5, the raw materials are in the following proportions by weight: 0.1-5 parts Phyllanthus emblica extract, 0.1-5 parts Morus alba extract, 0.05-5 parts concentrated material, 0.001-2 parts sweetener, 0.001-1 part acidity regulator, 0-0.05 parts excipients, and 50-500 parts water. The excipients may be added or omitted depending on the functional requirements.

[0029] Preferably, in step 5, the amount of amla extract added can be a fixed value or any range such as 0.2 parts, 0.3 parts, 0.5 parts, 0.8 parts, 1 part, 2 parts, 3 parts, or 4 parts.

[0030] Preferably, in step 5, the amount of mulberry extract added can be a fixed value or any range such as 0.2 parts, 0.3 parts, 0.5 parts, 0.8 parts, 1 part, 2 parts, 3 parts, or 4 parts.

[0031] Preferably, in step 5, the amount of concentrated material added can be a fixed value or any range such as 0.08 parts, 0.1 parts, 0.2 parts, 0.3 parts, 0.5 parts, 0.8 parts, 1 part, 2 parts, 3 parts, or 4 parts.

[0032] Preferably, in step 5, the amount of sweetener added can be a fixed value or any range such as 0.002 parts, 0.005 parts, 0.01 parts, 0.05 parts, 0.1 parts, 0.5 parts, 1 part, or 1.5 parts.

[0033] Preferably, in step 5, the amount of acidity regulator added can be a fixed value or any range such as 0.002 parts, 0.005 parts, 0.01 parts, 0.05 parts, 0.1 parts, 0.2 parts, 0.5 parts, or 1 part.

[0034] Preferably, in step 5, the amount of excipients added can be a fixed value or any range such as 0.00002 parts, 0.00005 parts, 0.00008 parts, 0.0001 parts, 0.0005 parts, 0.001 parts, 0.01 parts, or 0.05 parts.

[0035] Preferably, in step 5, the excipients include at least one of L-carnitine, tea polyphenols, preservatives, flavorings, and colorings.

[0036] Preferably, in step 5, the sweetener includes at least one of mogroside, xylitol, sorbitol, maltitol, sucrose, and glucose.

[0037] Preferably, in step 5, the acidity regulator includes at least one of citric acid, tartaric acid, malic acid, and lactic acid.

[0038] Preferably, in step 5, the amount of water added can be a fixed value or any range such as 60 parts, 80 parts, 90 parts, 100 parts, 200 parts, 300 parts, 400 parts, or 500 parts.

[0039] Preferably, in step 5, the preparation method of the amla extract includes: washing and drying amla, crushing it, adding an ethanol aqueous solution, heating and stirring, cooling, filtering, and concentrating to obtain the amla extract.

[0040] Preferably, the concentration of the ethanol-water solution is 30-50 wt%. The mass ratio of the ethanol-water solution to Phyllanthus emblica is 10-80:1. The heating and stirring temperature is 40-60℃, and the time is 2-6 hours. Concentration is carried out under reduced pressure to remove ethanol and some water.

[0041] Preferably, in step 5, the method for preparing the mulberry extract includes: washing and drying the mulberries, crushing them, adding water, heating and stirring, cooling, filtering, and concentrating to obtain the mulberry extract.

[0042] Preferably, the mass of water added is 10-50 times that of the mulberries. The heating and stirring temperature is 50-65℃, and the time is 2-7 hours. Concentration is carried out by vacuum concentration and removal of some water.

[0043] Preferably, in step 5, the aging process is carried out at room temperature for 0.5-2 hours.

[0044] The present invention also provides a fermented beverage prepared according to the preparation method described herein.

[0045] The beneficial effects of this invention are:

[0046] This invention uses wolfberry, cassia seed, burdock root, mulberry bark, and codonopsis as raw materials. These are boiled in water, then inoculated with specific strains of *Candida krusei* CICC 31807, *Saccharomyces cerevisiae* CICC 31105, and *Bifidobacterium animalis* CICC 6165 in a ratio of (2-5):1:(2.5-6) ​​for fermentation. After post-processing, a concentrated material is obtained. This concentrated material is then combined with *Phyllanthus emblica* extract, mulberry extract, sweetener, acidity regulator, excipients, and water to prepare a fermented beverage. The beverage prepared by this invention exhibits good stability, maintaining good stability even after exposure to high or low temperatures. It has a rich aroma, no bitterness, a smooth swallowing experience, a pleasant, refreshing, and delicate taste, and provides a superior overall experience.

[0047] This invention involves freeze-drying the concentrate and then conducting liver protection experiments. The tests show that the concentrated intermediate product prepared by this invention can effectively reduce the levels of aspartate aminotransferase (AST), malondialdehyde (MDA), and alanine aminotransferase (ALT), thus providing fermented beverages with a better liver protection effect.

[0048] This invention uses a specific fermentation strain combined with Candida cruzie CICC 31807, Saccharomyces cerevisiae CICC 31105 and Bifidobacterium animalis CICC 6165 for fermentation, which can effectively enhance the liver-protective effect of the intermediate product (the dried product of the concentrated material).

[0049] This invention uses Saccharomyces cerevisiae CICC 31105 as a base, and combines it with Candida cruzie CICC 31807 and Bifidobacterium animalis CICC 6165. The combination of Candida cruzie CICC 31807 and Bifidobacterium animalis CICC 6165 produces a synergistic effect in reducing the levels of aspartate aminotransferase, malondialdehyde, and alanine aminotransferase in the liver, thus giving the beverage a better liver-protecting effect.

[0050] This invention uses Saccharomyces cerevisiae CICC 31105 as a base, and combines Candida albicans CICC 31807 and Bifidobacterium animalis CICC 6165 in a ratio of (2-5):(2.5-6) ​​(that is, the ratio of the three is (2-5):1:(2.5-6)). The resulting intermediate product has a better liver-protecting effect, which makes the beverage have a better liver-protecting effect.

[0051] In the preparation process of this invention, the combination of cassia seed and burdock root creates a synergistic effect, giving the intermediate product a better effect in reducing the levels of aspartate aminotransferase, malondialdehyde, and alanine aminotransferase in the liver, thus making the beverage more liver-protective. Furthermore, a mass ratio of cassia seed to burdock root of 1:0.7-1.2 yields an even more effective intermediate product, giving the beverage a better overall effect. Detailed Implementation

[0052] The present invention will be described below with reference to specific embodiments. These specific embodiments are for illustrative purposes only and are not intended to limit the present invention.

[0053] The preparation method of amla extract includes: washing and drying amla, crushing it to a particle size of less than 1 cm, adding 22.4 times its mass of 45 wt% ethanol aqueous solution, heating at 50°C and stirring for 5 hours, cooling to 35°C, filtering with a 0.5 μm pore size filter membrane to obtain filtrate, and then concentrating under reduced pressure to 10.5% of the filtrate mass to obtain amla extract.

[0054] The preparation method of mulberry extract includes: washing and drying mulberries, crushing them to a particle size of less than 1 cm, adding 24.5 times their weight of water, heating at 60°C and stirring for 5.8 hours, cooling to 40°C, filtering through a 0.5 μm pore size filter membrane to obtain filtrate, and then concentrating under reduced pressure to 10.9% of the filtrate mass to obtain mulberry extract.

[0055] Preparation of fermented beverages:

[0056] 1. Beverage 1

[0057] Step 1: According to the following mass proportions: 0.5 parts wolfberry, 1 part cassia seed, 1.1 parts burdock root, 0.6 parts mulberry bark, and 0.4 parts codonopsis; wash and dry the wolfberry, cassia seed, burdock root, mulberry bark, and codonopsis, then crush them to a particle size of less than 1 cm to obtain crushed material; the mass ratio of cassia seed to burdock root is 1:1.1.

[0058] Step 2: Add 26 times the weight of water to the crushed material and boil it, then cool it to room temperature to obtain a mixture;

[0059] Step 3: Mix the mixture with 3.45% glucose by mass to obtain the treatment solution, inoculate with fermentation bacteria, and anaerobic ferment at 40°C for 8.5 days. Sterilize to obtain the fermentation product.

[0060] The fermentation strain consisted of *Candida krusei* CICC 31807, *Saccharomyces cerevisiae* CICC 31105, and *Bifidobacterium animalis* CICC 6165 in a ratio of 3.5:1:4.5. The inoculum size of the fermentation strain was 4.40 × 10⁻⁶ cells / mL of treatment solution. 9 CFU;

[0061] Step 4: Filter the fermentation material through a 0.5μm pore size filter membrane and concentrate it under reduced pressure to 11.25% of the fermentation material mass to obtain concentrated material;

[0062] Step 5: Add 1.2 parts of amla extract, 1.5 parts of mulberry extract, 1.2 parts of xylitol (sweetener), 0.1 parts of citric acid (acidity regulator), 0.006 parts of excipients (0.001 parts of tea polyphenols and 0.005 parts of potassium sorbate) and water (to a total of 100 parts) to 2 parts of concentrated material, stir well, sterilize, and let stand at room temperature for 1.2 hours to obtain the beverage intermediate;

[0063] Step 6: Fill and package the beverage intermediate to obtain the fermented beverage.

[0064] 2. Beverages

[0065] Step 1: According to the following mass proportions: 0.4 parts wolfberry, 1.2 parts cassia seed, 0.9 parts burdock root, 0.8 parts mulberry bark, and 0.3 parts codonopsis; wash and dry the wolfberry, cassia seed, burdock root, mulberry bark, and codonopsis, then crush them to a particle size of less than 1 cm to obtain crushed material; the mass ratio of cassia seed to burdock root is 1:0.75;

[0066] Step 2: Add 20 times the weight of water to the crushed material and boil it, then cool it to room temperature to obtain a mixture;

[0067] Step 3: Mix the mixture with 3.72% glucose by mass to obtain the treatment solution, inoculate with fermentation bacteria, and anaerobic ferment at 38°C for 9 days. Sterilize to obtain the fermentation product.

[0068] The fermentation strain consisted of *Candida krusei* CICC 31807, *Saccharomyces cerevisiae* CICC 31105, and *Bifidobacterium animalis* CICC 6165 in a ratio of 2:1:6. The inoculum size of the fermentation strain was 4.52 × 10⁻⁶ per mL of treatment solution. 9 CFU;

[0069] Step 4: Filter the fermentation material through a 0.5μm pore size filter membrane and concentrate it under reduced pressure to 11.39% of the fermentation material mass to obtain concentrated material;

[0070] Step 5: Add 1.5 parts of amla extract, 1.6 parts of mulberry extract, 1.18 parts of xylitol (sweetener), 0.12 parts of citric acid (acidity regulator), 0.006 parts of excipients (0.0012 parts of tea polyphenols and 0.0055 parts of potassium sorbate) and water (to a total of 100 parts) to 1.8 parts of concentrated material. Stir well, sterilize, and let stand at room temperature for 1.5 hours to obtain the beverage intermediate.

[0071] Step 6: Fill and package the beverage intermediate to obtain the fermented beverage.

[0072] 3. Beverages

[0073] The difference from beverage 1 is as follows: Step 1: According to the mass parts: 0.5 parts wolfberry, 2.1 parts burdock root, 0.6 parts mulberry bark and 0.4 parts codonopsis; wash and dry wolfberry, cassia seed, burdock root, mulberry bark and codonopsis, and then crush them to a particle size of less than 1cm to obtain crushed material; everything else is the same.

[0074] 4. Beverages

[0075] The difference from beverage 1 is as follows: Step 1: According to the mass parts: 0.5 parts wolfberry, 2.1 parts cassia seed, 0.6 parts mulberry bark and 0.4 parts codonopsis; wash and dry wolfberry, cassia seed, burdock root, mulberry bark and codonopsis, and then crush them to a particle size of less than 1cm to obtain crushed material; everything else is the same.

[0076] 5. Beverages

[0077] The difference from beverage 1 is as follows: Step 1: According to the mass parts: 0.5 parts wolfberry, 0.3 parts cassia seed, 1.8 parts burdock root, 0.6 parts mulberry bark and 0.4 parts codonopsis; wash and dry wolfberry, cassia seed, burdock root, mulberry bark and codonopsis, and then crush them to a particle size of less than 1cm to obtain crushed material; the mass ratio of cassia seed to burdock root is 1:6; everything else is the same.

[0078] 6. Beverages

[0079] The difference from beverage 1 is as follows: Step 1: According to the mass parts: 0.5 parts wolfberry, 1.7 parts cassia seed, 0.4 parts burdock root, 0.6 parts mulberry bark and 0.4 parts codonopsis; wash and dry wolfberry, cassia seed, burdock root, mulberry bark and codonopsis, and then crush them to a particle size of less than 1cm to obtain crushed material; the mass ratio of cassia seed to burdock root is 1:0.23; everything else is the same.

[0080] 7. Beverages

[0081] Step 1: According to the following mass proportions: 0.5 parts wolfberry, 1 part cassia seed, 1.1 parts burdock root, 0.6 parts mulberry bark, and 0.4 parts codonopsis; wash and dry the wolfberry, cassia seed, burdock root, mulberry bark, and codonopsis, then crush them to a particle size of less than 1 cm to obtain crushed material; the mass ratio of cassia seed to burdock root is 1:1.1.

[0082] Step 2: Add 26 times the weight of water to the crushed material and boil it, then cool it to room temperature to obtain a mixture;

[0083] Step 3: Filter the mixture through a 0.5 μm pore size filter membrane and concentrate it under reduced pressure to 11.25% of the mixture mass to obtain concentrated material;

[0084] Step 4: Add 1.2 parts of amla extract, 1.5 parts of mulberry extract, 1.2 parts of xylitol (sweetener), 0.1 parts of citric acid (acidity regulator), 0.006 parts of excipients (0.001 parts of tea polyphenols and 0.005 parts of potassium sorbate) and water (to a total of 100 parts) to 2 parts of concentrated material, stir well, sterilize, and let stand at room temperature for 1.2 hours to obtain the beverage intermediate;

[0085] Step 5: Fill and package the beverage intermediate to obtain the fermented beverage.

[0086] 8. Beverages

[0087] The difference from beverage 1 is as follows: Step 3: The mixture is stirred with 3.45% glucose by mass to obtain a treatment solution. Fermentation bacteria are inoculated, and the mixture is anaerobic fermented at 40℃ for 8.5 days. The solution is then sterilized to obtain the fermented product. The fermentation bacteria consist of *Saccharomyces cerevisiae* CICC 31105 and *Bifidobacterium animalis* CICC 6165 in a ratio of 1:8. The inoculation amount of fermentation bacteria in each mL of treatment solution is 4.40 × 10⁻⁶. 9 cfu; everything else is the same.

[0088] 9. Beverages

[0089] The difference from beverage 1 is as follows: Step 3: The mixture is stirred with 3.45% glucose by mass to obtain a treatment solution. Fermentation bacteria are inoculated, and the mixture is anaerobic fermented at 40℃ for 8.5 days. The solution is then sterilized to obtain the fermented product. The fermentation bacteria consist of *Candida cruzii* CICC 31807 and *Saccharomyces cerevisiae* CICC 31105 in a ratio of 8:1. The inoculation amount of the fermentation bacteria in each mL of treatment solution is 4.40 × 10⁻⁶. 9 cfu; everything else is the same.

[0090] 10. Beverage bacterial count (per 10%)

[0091] The difference from beverage 1 is as follows: Step 3: The mixture is stirred with 3.45% glucose by mass to obtain a treatment solution. Fermentation bacteria are inoculated, and the mixture is anaerobic fermented at 40℃ for 8.5 days. The solution is then sterilized to obtain the fermented product. The fermentation bacteria consist of *Candida krusei* CICC 31807, *Saccharomyces cerevisiae* CICC 31105, and *Bifidobacterium animalis* CICC 6165 in a ratio of 1:1:7. The inoculation amount of the fermentation bacteria in each mL of treatment solution is 4.40 × 10⁻⁶. 9 cfu; everything else is the same.

[0092] 11. Beverage bacterial count

[0093] The difference from beverage 1 is as follows: Step 3: The mixture is stirred with 3.45% glucose by mass to obtain a treatment solution. Fermentation bacteria are inoculated, and the mixture is anaerobic fermented at 40℃ for 8.5 days. The solution is then sterilized to obtain the fermented product. The fermentation bacteria consist of *Candida krusei* CICC 31807, *Saccharomyces cerevisiae* CICC 31105, and *Bifidobacterium animalis* CICC6165 in a ratio of 7.2:1:0.8. The inoculation amount of the fermentation bacteria in each mL of treatment solution is 4.40 × 10⁻⁶. 9 cfu; everything else is the same.

[0094] II. Performance Testing

[0095] 1. Stability

[0096] The prepared beverage was subjected to high-temperature and low-temperature stability tests. High-temperature stability test: The prepared beverage was sealed and stored at 55℃ for 7 days, then naturally cooled to room temperature, and its appearance was observed (whether there was any precipitation or flocculent matter). Low-temperature stability test: The prepared beverage was sealed and stored at -10℃ for 7 days, then naturally warmed to room temperature, and its appearance was observed (whether there was any precipitation or flocculent matter). Specific test results are shown in Table 1.

[0097] Table 1: Stability Tests

[0098]

[0099] As can be seen from the test results in Table 1, the beverage prepared by this invention has good stability and maintains good stability even after being subjected to high temperature of 55℃ or low temperature of -10℃.

[0100] 2. Sensory experience test

[0101] Fifteen volunteers (aged 30-55, 7 males and 8 females) were recruited to rate and evaluate the prepared beverages 1-2 and 711. Evaluation indicators included aroma, taste, and texture. Aroma was defined as a fresh, herbal fragrance; the stronger the aroma, the higher the score, with a total score of 10. Taste was defined as the absence of bitterness and smoothness of swallowing; the higher the smoothness of swallowing, the higher the score, with a total score of 10. Texture was defined as refreshing and delicate; the higher the refreshingness and the more delicate the texture, the higher the score, with a total score of 10. The highest and lowest scores were removed, and the average of all indicators was taken, rounded to two decimal places. The results are shown in Table 2.

[0102] Table 2: Sensory Experience Test

[0103]

[0104] As can be seen from the test results in Table 2, the beverage prepared by this invention has a rich aroma, good taste, and good condition.

[0105] 3. Liver protection effect test

[0106] Healthy male SPF-grade C57BL / 6 mice weighing 20±2g were used as experimental subjects. The concentrated material prepared in beverage 1-11 was freeze-dried at -42℃ to a water content of 2.41% as the drug sample for testing.

[0107] The specific experimental procedure was as follows: Mice were acclimatized for 4 days and then randomly divided into 13 groups (beverage groups 1-11, blank group, and model group) according to their body weight, with 12 mice in each group; except for the blank group, the mice in the other groups were gavaged with 0.055 mL / 10 g / day of 52° liquor, while the blank group was gavaged with an equal volume of physiological saline for 2 consecutive weeks; starting from the first day of liquor gavage, the mice in beverage groups 1-11 were gavaged with 0.12 mL / 10 g / day of the corresponding drug sample's physiological saline solution (the drug sample dosage was 0.8 g / kg / day, diluted with physiological saline to obtain the solution), while the blank group and model group were gavaged with an equal volume of physiological saline every day; during the experiment, the mice in each group had free access to food and water.

[0108] Mice were sacrificed 24 hours after the last administration. Liver homogenate was collected, and the levels of aspartate aminotransferase (AST), malondialdehyde (MDA), and alanine aminotransferase (ALT) were measured. The average values ​​were taken. The results are shown in Table 3.

[0109] Table 3: Content Test

[0110]

[0111] The test values ​​in Table 3 were converted to obtain the reduction values ​​Δ of each parameter compared to the model group. The results are shown in Table 4.

[0112] Table 4: Decrease values ​​Δ for each parameter

[0113]

[0114] Based on the test results in Table 3-4, it can be seen that after alcohol gavage treatment, the levels of aspartate aminotransferase (AST), malondialdehyde (MDA), and alanine aminotransferase (ALT) in the liver of mice were significantly increased. This invention uses wolfberry, cassia seed, burdock root, mulberry bark, and codonopsis as raw materials, boils them in water, and then inoculates them with specific strains of Candida albicans CICC31807, Saccharomyces cerevisiae CICC 31105, and Bifidobacterium animalis CICC 6165 in a ratio of (2-5):1:(2.5-6) ​​for fermentation. The resulting intermediate product (the dried product of the concentrated material) after post-processing effectively reduces the levels of AST, MDA, and ALT, giving the fermented beverage a better liver-protective effect.

[0115] Based on the test results of beverages 1 and 7, it is known that the present invention uses a specific fermentation strain combined with Candida cruzie CICC31807, Saccharomyces cerevisiae CICC 31105 and Bifidobacterium animalis CICC 6165 for fermentation, which can effectively enhance the liver-protective effect of the intermediate product (the dried product of the concentrated material).

[0116] Based on beverages 1 and 8-9, it can be seen that, while maintaining the total amount of fermentation bacteria and the amounts of *Candida cruzi* CICC 31807 and *Bifidobacterium animalis* CICC 6165 unchanged, the levels of aspartate aminotransferase (AST), malondialdehyde (MDA), and alanine aminotransferase (ALT) in the livers of mice obtained by omitting either *Candida cruzi* CICC 31807 or *Bifidobacterium animalis* CICC 6165 during fermentation in beverage 8-9 were higher than those in beverage 1. This means that by using *Saccharomyces cerevisiae* CICC 31105, the combination of *Candida cruzi* CICC 31807 and *Bifidobacterium animalis* CICC 6165 results in a synergistic effect, reducing the levels of AST, MDA, and ALT in the liver, thus providing the beverage with better liver-protective effects.

[0117] Based on beverages 1 and 10-11, it can be seen that, while maintaining the total amount of fermentation bacteria and the amounts of *Candida cruzi* CICC31807 and *Bifidobacterium animalis* CICC 6165 unchanged, changing the amounts of *Candida cruzi* CICC 31807 and *Bifidobacterium animalis* CICC 6165 resulted in higher levels of aspartate aminotransferase (AST), malondialdehyde (MDA), and alanine aminotransferase (ALT) in the livers of mice fermented with beverage 10-11 compared to beverage 1. This means that, based on *Saccharomyces cerevisiae* CICC31105, using a ratio of *Candida cruzi* CICC 31807 and *Bifidobacterium animalis* CICC 6165 of (2-5):(2.5-6) ​​(i.e., a ratio of (2-5):1):(2.5-6)) yields a more effective liver-protective intermediate product, thus enhancing the beverage's liver-protective properties.

[0118] According to the test results of beverages 1, 3-6, the combination of cassia seed and burdock root in the preparation process of this invention produces a synergistic effect, giving the intermediate product a better effect in reducing the levels of aspartate aminotransferase, malondialdehyde, and alanine aminotransferase in the liver, thus making the beverage have a better liver-protective effect. Furthermore, an intermediate product with a mass ratio of cassia seed to burdock root of 1:0.7-1.2 is even more effective, giving the beverage a better overall effect.

[0119] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A method for preparing a fermented beverage, characterized in that, Includes the following steps: Step 1: According to the following weight parts: 0.5 parts wolfberry, 1 part cassia seed, 1.1 parts burdock root, 0.6 parts mulberry bark and 0.4 parts codonopsis; wash and dry the wolfberry, cassia seed, burdock root, mulberry bark and codonopsis, and then crush them to a particle size of less than 1cm to obtain crushed material; Step 2: Add 26 times the weight of water to the crushed material and boil it, then cool it to room temperature to obtain a mixture; Step 3: Mix the mixture with 3.45% glucose by mass to obtain the treatment solution, inoculate with fermentation bacteria, and anaerobic ferment at 40°C for 8.5 days. Sterilize to obtain the fermentation product. The fermentation strain consisted of *Candida krusei* CICC 31807, *Saccharomyces cerevisiae* CICC 31105, and *Bifidobacterium animalis* CICC 6165 in a ratio of 3.5:1:4.

5. The inoculum size of the fermentation strain was 4.40 × 10⁻⁶ cells / mL of treatment solution. 9 CFU; Step 4: Filter the fermentation material through a 0.5μm pore size filter membrane and concentrate it under reduced pressure to 11.25% of the fermentation material mass to obtain concentrated material; Step 5: Add 1.2 parts of Phyllanthus emblica extract, 1.5 parts of mulberry extract, 1.2 parts of xylitol (sweetener), 0.1 parts of citric acid (acidity regulator), 0.006 parts of excipients, and water to 2 parts of concentrated material to a total of 100 parts. Stir well, sterilize, and let stand at room temperature for 1.2 hours to obtain the beverage intermediate. The excipients are 0.001 parts of tea polyphenols and 0.005 parts of potassium sorbate. Step 6: Fill and package the beverage intermediate to obtain the fermented beverage.

2. The preparation method according to claim 1, characterized in that, The preparation method of the amla extract includes: washing and drying amla, crushing it, adding an ethanol aqueous solution, heating and stirring, cooling, filtering, and concentrating to obtain the amla extract.

3. The preparation method according to claim 1, characterized in that, The method for preparing the mulberry extract includes: washing and drying mulberries, crushing them, adding water, heating and stirring, cooling, filtering, and concentrating to obtain the mulberry extract.

4. A fermented beverage prepared by the preparation method according to any one of claims 1-3.

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