Liver-protecting beverage containing probiotics and preparation method of liver-protecting beverage

Abstract

The invention relates to the technical field of probiotic fermented beverages, in particular to a liver-protecting beverage containing probiotics and a preparation method thereof, and the liver-protecting beverage containing probiotics is prepared from 20-30 parts by weight of probiotic freeze-dried powder, 30-50 parts by weight of radix puerariae, 5-15 parts by weight of semen hoveniae, 5-15 parts by weight of natto, 10-15 parts by weight of yeast and 5000-10000 parts by weight of water. The preparation method comprises the following steps: S1, weighing the radix puerariae, the hovenia dulcis thunb and the natto for later use; s2, respectively washing, drying, crushing and mixing the radix puerariae and the semen hoveniae to obtain mixed powder, and extracting and concentrating the mixed powder into extract; s3, grinding natto into powder, adding the powder into the extract, uniformly mixing, drying, crushing, sieving, adding water and yeast for fermentation, and after fermentation is finished, filtering and sterilizing to obtain fermentation liquor; s4, adding probiotic freeze-dried powder into the fermentation liquid obtained in S3, and performing secondary fermentation; and S5, taking the clarified liquid for filling. According to the invention, the absorption of ethanol is reduced, the activity of acetaldehyde dehydrogenase (ALDH) and ethanol dehydrogenase (ADH) is improved, and the metabolism of ethanol in the liver is promoted, so that the purpose of protecting the liver is achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of probiotic fermented beverages, in particular to a probiotic-containing liver-protecting beverage and a preparation method thereof. Background Art

[0002] Alcohol is very common in people's daily lives. Moderate drinking can relieve tension and fatigue and speed up the body's metabolism. However, if you drink too much, most of the ethanol that enters the body will be metabolized through the liver, which will damage the structure of liver cells, thereby disrupting normal cell metabolism and producing cytotoxic metabolites. Alcohol poisoning may occur and even cause coma and shock. It can be seen that alcohol has had a significant impact on people's health.

[0003] After alcohol enters the body, the vast majority of it enters the liver through the bloodstream for metabolism. Two key enzymes, alcohol dehydrogenase (EC.1.1.1.1, abbreviated ADH) and acetaldehyde dehydrogenase (EC1.2.1.n, abbreviated ALDH), degrade it into acetic acid. Under aerobic conditions, acetic acid enters the tricarboxylic acid cycle or other pathways, where it is completely oxidized into water and carbon dioxide, releasing energy. ADH is a zinc-containing enzyme (Zn-enzyme) that depends on coenzyme I (NAD+). ALDH requires either coenzyme I (NAD+) or coenzyme II (NADP+) to function.

[0004] Under normal circumstances, the body's enzymes and their activity are maintained at a certain level to meet the body's needs. However, when drinking excessive amounts of alcohol, these enzymes and their activity are insufficient to break down the alcohol, causing liver damage. Therefore, we aim to provide a liver-protecting beverage containing probiotics that can, to a certain extent, increase the activity of these key enzymes and reduce the harmful effects of alcohol on the liver. Summary of the Invention

[0005] In view of the shortcomings of the prior art, the object of the present invention is to provide a liver-protecting beverage containing probiotics and a preparation method thereof.

[0006] The following explains the content and its effects: A probiotic-containing liver-protecting beverage comprises 20-30 parts by weight of probiotic freeze-dried powder, 30-50 parts by weight of kudzu root, 5-15 parts by weight of Hovenia dulcis fruit, 5-15 parts by weight of natto, 10-15 parts by weight of yeast, and 5,000-10,000 parts by weight of water.

[0007] The probiotic freeze-dried powder includes Lactobacillus casei freeze-dried powder, Lactobacillus acidophilus freeze-dried powder, Bifidobacterium longum freeze-dried powder, and Bifidobacterium infantis freeze-dried powder, and the ratio thereof is 1:(1~2):(1~2):(1~2).

[0008] A method for preparing a liver-protecting beverage containing probiotics comprises the following steps: S1. Weigh Pueraria root, Hovenia dulcis fruit, and Natto and set aside. S2, washing, drying, and crushing Pueraria root and Hovenia dulcis fruit respectively, mixing to obtain a mixed powder, and extracting and concentrating to obtain an extract; S3, grinding the natto into powder and adding the extract, mixing, drying, crushing and sieving, adding water and yeast to ferment, and after fermentation, filtering and sterilizing to obtain a fermentation liquid; S4, adding probiotic freeze-dried powder to the fermentation liquid obtained in S3 to perform secondary fermentation; S5, taking the clarified liquid for filling.

[0009] In said S2, when extracting and concentrating to obtain an extract, add ethanol aqueous solution, reflux extract at 85°C to 95°C for 4 to 6 hours, filter out the extract, re-extract the residue, and combine the filtrates; the filtrates are vacuum concentrated at 85°C to obtain an extract.

[0010] The concentration of the ethanol aqueous solution is 50% to 70%, and the ratio of the weight of the ethanol aqueous solution to the weight of the mixed powder is (6-8):1.

[0011] In S2 and S3, the drying temperature is 50-55°C.

[0012] In S3, water and yeast are added for fermentation at a temperature of 28-36° C. for 5-14 days.

[0013] The temperature of the secondary fermentation in S4 is 28-30° C. and the time is 5-7 days.

[0014] The invention discloses a method for drinking a liver-protecting beverage containing probiotics, which comprises directly adding the liver-protecting beverage containing probiotics into wine and drinking the beverage together with the wine.

[0015] In summary, the present invention includes at least one of the following beneficial technical effects: 1. Probiotic fermentation of Pueraria root, Hovenia dulcis fruit, and Natto can significantly increase the content of active ingredients such as puerarin, daidzein, flavonoids, and polyphenols, thereby promoting the body's breakdown and absorption of alcohol and enhancing the beverage's liver-protective properties. Related research shows that puerarin and daidzein can reduce ethanol absorption, increase the activity of acetaldehyde dehydrogenase (ALDH) and alcohol dehydrogenase (ADH), and promote ethanol metabolism in the body, thereby achieving liver protection. Flavonoids and polyphenols have strong free radical scavenging abilities, which can reduce liver damage.

[0016] 2. The probiotic fermentation process produces lactic acid and other flavor components, giving the beverage a unique fermented food aroma. This can improve the bitterness of traditional Chinese medicine, enhance the taste of the liver-protecting beverage, and nourish the spleen, stomach, liver, and kidneys. This liver-protecting beverage can also help adjust the taste of baijiu (white liquor), making it more palatable. Therefore, it can be added to baijiu and then consumed together.

[0017] 3. Liver-protecting beverages containing probiotics are fermented using a combination of bacterial strains. Different microorganisms have different enzyme systems. Using yeast and probiotic fermentation can diversify and complement the enzyme systems in the system, improving the biotransformation efficiency of complex traditional Chinese medicines. Secondary fermentation can also fully decompose, dissolve, and utilize the active ingredients and nutrients in plant tissues, improving the comprehensive utilization and biotransformation rate of traditional Chinese medicines, thereby enhancing the liver-protecting ability of liver-protecting beverages. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a typical graph for evaluating the auxiliary protective efficacy of the liver protection drink in Example 1 against alcoholic liver injury; Figure 2 is the average brightness of zebrafish liver after treatment with the liver protection drink in Example 1; Figure 3 Comparison of the liver area of zebrafish treated with the liver protection drink in Example 1 with that of the model control group; Figure 4 This is a comparison of the delayed absorption area of zebrafish yolk sac after treatment with the liver-protecting drink in Example 1 and the model control group. DETAILED DESCRIPTION

[0019] The present invention is described in further detail below. Example 1

[0020] The preparation method of the liver-protecting beverage containing probiotics comprises the following steps: S1. Weigh 40 parts of Puerariae Radix, 10 parts of Hovenia Dulcis Fruit, and 10 parts of Natto and set aside. S2, Pueraria root and Hovenia dulcis fruit are washed, dried at 55°C, and crushed to obtain a mixed powder, to which a 60% (volume ratio) ethanol aqueous solution is added, with the weight ratio of the ethanol aqueous solution to the mixed powder being 7:1. The mixture is refluxed at 90°C for 5 hours, the extract is filtered, the residue is re-extracted, and the filtrates are combined; the filtrates are vacuum concentrated at 85°C to obtain an extract; S3, grinding the natto powder and adding the extract, mixing, drying at 55°C, crushing and sieving, adding 8000 parts of water and 12 parts of yeast (high-activity dry yeast purchased from Angel Yeast Co., Ltd.) and fermenting at a temperature of 30°C for 10 days. After fermentation, filtering and sterilizing to obtain a fermentation liquid; S4, adding 25 parts of probiotic freeze-dried powder (the probiotic freeze-dried powder includes Lactobacillus casei freeze-dried powder, Lactobacillus acidophilus freeze-dried powder, Bifidobacterium longum freeze-dried powder, and Bifidobacterium infantis freeze-dried powder, and the ratio is 1:1:1:1, and the freeze-dried powder is self-developed by the applicant company) to the fermentation liquid obtained in S3, and performing secondary fermentation. The secondary fermentation temperature is 28°C and the time is 7 days; S5, taking the clarified liquid for filling. Example 2-3

[0021] The difference between Example 2-3 and Example 1 is that the weight parts of the probiotic freeze-dried powder added are different. The weight parts added in Example 2 is 20, and the weight parts added in Example 3 is 30. Examples 4-5

[0022] The difference between Example 4-5 and Example 1 is that the weight parts of Pueraria root added are different. The weight parts added in Example 4 is 30, and the weight parts added in Example 5 is 50. Examples 6-7

[0023] The difference between Example 6-7 and Example 1 is that the weight parts of Hovenia dulcis added are different, that is, 5 weight parts are added in Example 6, and 15 weight parts are added in Example 7. Examples 8-9

[0024] The difference between Example 8-9 and Example 1 is that the weight parts of natto added are different. In Example 8, 5 weight parts are added, while in Example 9, 15 weight parts are added. Examples 10-11

[0025] The difference between Example 10-11 and Example 1 is that the weight parts of yeast added are different. In Example 10, the weight parts added are 10, and in Example 11, the weight parts added are 15. Examples 12-13

[0026] The difference between Example 12-13 and Example 1 is that the weight parts of water added are different. The weight parts added in Example 12 is 5000, and the weight parts added in Example 13 is 10000. Example 14

[0027] The difference between Example 14 and Example 1 is that the ratio of each bacteria in the probiotic freeze-dried powder is different. In Example 14, the ratio of Lactobacillus casei freeze-dried powder, Lactobacillus acidophilus freeze-dried powder, Bifidobacterium longum freeze-dried powder, and Bifidobacterium infantis freeze-dried powder is 1:2:2:2. Examples 15-16

[0028] The difference between Example 15-16 and Example 1 is that the ethanol aqueous solution used in S2 is different. The ethanol aqueous solution in Example 15 is 50% (volume ratio), and the ethanol aqueous solution in Example 16 is 70% (volume ratio). Examples 17-18

[0029] The difference between Examples 17-18 and Example 1 is that when S2 is extracted and concentrated into an extract, the extraction temperature and time after adding the ethanol aqueous solution are different. Example 17 is refluxed at 85°C for 6 hours, and Example 18 is refluxed at 95°C for 4 hours. Example 19

[0030] The difference between Example 19 and Example 1 is that the drying temperatures in S2 and S3 are different, and the drying temperature in Example 19 is 50°C. Examples 20-21

[0031] The difference between Examples 20-21 and Example 1 is that the fermentation temperature and time when S3 is added to the yeast for fermentation are different. The fermentation temperature of Example 20 is 28°C and the time is 14 days; the fermentation temperature of Example 21 is 36°C and the time is 5 days. Example 22

[0032] The difference between Example 22 and Example 1 is that the temperature and time of the secondary fermentation in S4 are different. The fermentation temperature of Example 22 is 30° C. and the time is 5 days. Examples 23-24

[0033] The difference between Examples 23-24 and Example 1 is that the ratio of the weight of the ethanol aqueous solution to the weight of the mixed powder in S2 is different, that is, 6:1 in Example 23 and 8:1 in Example 24. Comparative Example 1

[0034] The difference between Comparative Example 1 and Example 1 is that in Comparative Example 1, no probiotic freeze-dried powder was added and no second fermentation was performed. Comparative Example 2

[0035] The difference between Comparative Example 2 and Example 1 is that in Comparative Example 2, no yeast was added and the first fermentation was not performed. Comparative Example 3

[0036] The difference between Comparative Example 3 and Example 1 is that in Comparative Example 3, Pueraria root is not added. Comparative Example 4

[0037] The difference between Comparative Example 4 and Example 1 is that in Comparative Example 4, Hovenia dulcis fruit was not added. Comparative Example 5

[0038] The difference between Comparative Example 5 and Example 1 is that natto is not added in Comparative Example 5. Application Example 1

[0039] 20 mL of the probiotic-containing liver-protecting beverage obtained in Example 1 was added to 20 mL of white wine, and the mixture was consumed together. Application Example 2

[0040] After drinking the liquor, 20 mL of the liver-protecting beverage containing probiotics obtained in Example 1 was also consumed. Experimental results analysis and discussion

[0041] The liver protection drink prepared in Example 1 was tested for its protective effect against alcoholic liver damage.

[0042] Sample: The live bacteria liver-protecting beverage prepared in Example 1.

[0043] The positive control was: Metadoxine Capsules (hereinafter referred to as Metadoxine), white powder, batch number 230801, Zhejiang Zhenyuan Pharmaceutical Co., Ltd., the solvent was DMSO.

[0044] Experimental animals: Zebrafish were kept in aquaculture water at 28°C (water quality: 200 mg of instant sea salt was added to each liter of reverse osmosis water, conductivity was 450-550 uS / cm; pH was 6.5-8.5; hardness was 50-100 mg LCaCO3).

[0045] Instruments, consumables, and reagents: dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); precision electronic balance (CP214, OHAUS, USA); ultrasonic cleaning machine (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China); 6-well plate (Zhejiang Beilanbo Biotechnology Co., Ltd., China).

[0046] Dimethyl sulfoxide (DMSO, batch number 20220614, Sinopharm Chemical Reagent Co., Ltd., China); methylcellulose (batch number G2106167, Shanghai Aladdin Biochemical Technology Co., Ltd., China); anhydrous ethanol (batch number F2209028, Shanghai Aladdin Biochemical Technology Co., Ltd., China).

[0047] Test method: Maximum Test Concentration (MTC) Assay: Wild-type AB zebrafish, 3 days post-fertilization (dpf), were randomly selected and plated in 6-well plates, with 30 zebrafish treated in each well (experimental group). Samples were administered in water, along with normal and model control groups. Each well contained 3 mL of sample solution. Except for the normal control group, all other experimental groups were administered with anhydrous ethanol in water to establish an alcoholic liver injury model. After treatment at 28°C for 2 days, the MTC of the sample in the model zebrafish was measured.

[0048] Evaluation of the adjuvant protective efficacy against alcoholic liver injury: 3-day-old wild-type AB zebrafish were randomly selected and plated in 6-well plates, with 30 zebrafish treated in each well (experimental group). The samples were administered in water, along with the positive control metadoxine at a concentration of 149 μg / mL. A volume of 3 mL was also set up in each well for the normal and model control groups. All experimental groups, except the normal control group, were administered with anhydrous ethanol in water to establish an alcoholic liver injury model. After two days of treatment at 28°C, 10 zebrafish were randomly selected from each experimental group and photographed under a dissecting microscope. Data were analyzed and collected using NIS-Elements D3.20 advanced image processing software. Liver area, mean liver brightness, and yolk sac resorption delay area were analyzed. The adjuvant protective efficacy of the samples against alcoholic liver injury was evaluated using these indicators. Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software. P < 0.05 indicated statistical significance.

[0049] Test results: MTC: Under the experimental conditions, the MTC of the liver protection drink in Example 1 for auxiliary protection against alcoholic liver injury is 1.25 mL / mI.

[0050] Evaluation of auxiliary protective efficacy against alcoholic liver injury: Under the experimental conditions, the liver protection beverage of Example 1 has auxiliary protective efficacy against alcoholic liver injury, which is specifically manifested in reducing liver area, increasing the average brightness of the liver, and reducing the delayed area of yolk sac absorption. Figures 1 to 4 .

[0051] Comparing Examples 1-3 and Comparative Example 1, no probiotics were added to Comparative Example 1, and the final liver protection effect was significantly inferior to that of Example 1, and the flavor of the beverage was also far inferior to that of Example 1. This is mainly because the probiotic fermentation of Pueraria lobata, Hovenia dulcis, and Natto can significantly increase the content of effective ingredients such as puerarin, daidzein, and polyphenols, increase the activity of acetaldehyde dehydrogenase (ALDH) and alcohol dehydrogenase (ADH), and promote the metabolism of ethanol in the body, thereby achieving the purpose of protecting the liver, thereby promoting the decomposition and absorption of alcohol by the human body and enhancing the liver protection ability of the beverage. Flavonoids and polyphenols have a strong ability to scavenge free radicals, which can reduce liver damage. In addition, different microorganisms have different enzyme systems. Using yeast fermentation and probiotic fermentation can diversify and complement the enzyme systems of the system, and the biotransformation efficiency of traditional Chinese medicine with complex ingredients is higher. After secondary fermentation, the effective ingredients and nutrients in the plant tissue can be fully decomposed, dissolved, and utilized, thereby improving the comprehensive utilization rate and biotransformation rate of traditional Chinese medicine, thereby improving the liver protection ability of the liver protection beverage.

[0052] Sensory evaluation form

[0053] Comparing Example 1, Examples 4-7, and Comparative Examples 3-4, in Comparative Example 3, Pueraria root was not added, and the final liver protection effect and beverage flavor were far inferior to those in Example 1. In Comparative Example 4, Hovenia dulcis fruit was not added, which also had a relatively large impact on the liver protection effect and flavor.

[0054] Comparing Example 1, Examples 8-9, and Comparative Example 5, no natto was added in Comparative Example 5, which also affected the liver protection effect and the flavor of the drink.

[0055] Comparing Example 1, Examples 10-11, and Comparative Example 2, Comparative Example 2 did not use yeast for the first fermentation, which affected the activities of acetaldehyde dehydrogenase (ALDH) and alcohol dehydrogenase (ADH), and had a relatively obvious impact on the liver protection effect and beverage flavor.

[0056] Example 1 is applied to Application Example 1 and Application Example 2 respectively. Both methods can achieve good liver protection effects. What is more special is that in Application Example 1, adding the beverage to the white wine and drinking the two together will have a better flavor.

[0057] In particular, the solution of Example 1 reduces the absorption of ethanol, increases the activity of acetaldehyde dehydrogenase (ALDH) and alcohol dehydrogenase (ADH), and promotes the metabolism of ethanol in the body, thereby achieving the purpose of protecting the liver. Flavonoids and polyphenols have a strong ability to scavenge free radicals, which can reduce liver damage. During the probiotic fermentation process, the substances produced by the first fermentation and the substances produced by the second fermentation can produce flavor components such as lactic acid to give the beverage a special fermented food aroma, improve the bitter taste of Chinese medicinal materials, increase the taste of liver-protecting beverages, and at the same time nourish the spleen, stomach, liver and kidneys. The liver-protecting beverage can help adjust the taste of white wine and make it more palatable, so the beverage can be added to the white wine and mixed, and then drunk together. The liver-protecting beverage containing probiotics is fermented by a composite strain. Different microorganisms have different enzyme systems. Using yeast fermentation and probiotic fermentation can diversify and complement the system's enzyme system, and have higher efficiency in the biotransformation of traditional Chinese medicine with complex ingredients. After secondary fermentation, the effective ingredients and nutrients in plant tissues can be fully decomposed, dissolved and utilized, thereby improving the comprehensive utilization rate and biotransformation rate of traditional Chinese medicine, thereby improving the liver protection ability of liver protection beverages.

[0058] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make non-creative modifications to the present embodiment as needed, but as long as they are within the scope of the present application, they are protected by the patent law.

Claims

1. A liver-protecting beverage containing probiotics, characterized in that: The invention comprises 20-30 parts by weight of probiotic freeze-dried powder, 30-50 parts by weight of kudzu root, 5-15 parts by weight of hovenia dulcis fruit, 5-15 parts by weight of natto, 10-15 parts by weight of yeast and 5000-10000 parts by weight of water.

2. The liver-protecting beverage containing probiotics according to claim 1, characterized in that: The probiotic freeze-dried powder includes Lactobacillus casei freeze-dried powder, Lactobacillus acidophilus freeze-dried powder, Bifidobacterium longum freeze-dried powder, and Bifidobacterium infantis freeze-dried powder, and the ratio thereof is 1:(1~2):(1~2):(1~2).

3. A method for preparing the liver-protecting beverage containing probiotics according to claim 1, characterized in that: The following steps are involved: S1. Weigh Pueraria root, Hovenia dulcis fruit, and Natto and set aside. S2, washing, drying, and crushing Pueraria root and Hovenia dulcis fruit respectively, mixing to obtain a mixed powder, and extracting and concentrating to obtain an extract; S3, grinding the natto into powder and adding the extract, mixing, drying, crushing and sieving, adding water and yeast to ferment, and after fermentation, filtering and sterilizing to obtain a fermentation liquid; S4, adding probiotic freeze-dried powder to the fermentation liquid obtained in S3 to perform secondary fermentation; S5, taking the clarified liquid for filling.

4. The preparation method according to claim 3, characterized in that In said S2, when extracting and concentrating to obtain an extract, add ethanol aqueous solution, reflux extract at 85°C to 95°C for 4 to 6 hours, filter out the extract, re-extract the residue, and combine the filtrates; the filtrates are vacuum concentrated at 85°C to obtain an extract.

5. The preparation method according to claim 4, characterized in that The concentration of the ethanol aqueous solution is 50% to 70%, and the weight ratio of the ethanol aqueous solution to the mixed powder is (6-8):

1.

6. The preparation method according to claim 3, characterized in that In S2 and S3, the drying temperature is 50-55°C.

7. The preparation method according to claim 3, characterized in that In S3, water and yeast are added for fermentation at a temperature of 28-36° C. for 5-14 days.

8. The preparation method according to claim 3, characterized in that The temperature of the secondary fermentation in S4 is 28-30° C. and the time is 5-7 days.

9. A method for drinking a liver-protecting beverage containing probiotics, characterized in that: The probiotic-containing liver-protecting beverage according to claim 1 or 2 or the probiotic-containing liver-protecting beverage prepared by the preparation method according to any one of claims 3 to 8 is directly added to the wine and consumed together with the wine.

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