Preparation method of composition for dispelling effects of alcohol and protecting liver

Abstract

The invention discloses a preparation method of a composition for dispelling effects of alcohol and protecting liver, and belongs to the technical field of traditional Chinese medicines. The liver protection composition is prepared from the following raw materials: gynostemma pentaphyllum powder, honeysuckle flower powder, hawthorn fruit powder, green tea powder, lotus leaf powder, cassia seed powder and folium sennae powder. The preparation method of the liver protection composition comprises the following steps: enzymolysis, fermentation and freeze-drying. According to the invention, through an extraction method of firstly performing enzymolysis and then performing fermentation, various different types of active effective components are reserved as far as possible. Tests prove that the finally obtained composition has a statistical therapeutic effect on alcoholic liver injury.

Description

Technical Field

[0001] The present invention belongs to the technical field of health products, and particularly relates to a preparation method of an anti-alcoholism and liver-protecting composition. Background Art

[0002] The liver is an important metabolic and detoxifying organ of the human body. Under normal circumstances, it can efficiently process toxins, metabolic wastes, and the conversion and storage of nutrients, maintain the physiological functions of the human body, and play a variety of key roles, such as synthesizing proteins, regulating blood sugar, and storing vitamins and minerals. In addition, the liver is also involved in the metabolism of fats, sugars, and proteins, as well as hormone inactivation and drug metabolic transformation, which is crucial for maintaining the internal environment stability. However, when liver function is damaged, a series of serious health problems will occur. For example:

[0003] 1. Decreased detoxification ability: leading to the accumulation of toxins and harmful substances, and causing poisoning symptoms;

[0004] 2. Metabolic dysfunction: manifested as metabolic syndromes such as fatty liver, hyperglycemia, and hyperlipidemia;

[0005] 3. Reduced protein synthesis: causing edema, ascites, or bleeding tendency;

[0006] 4. Risk of disease progression: may develop into liver cirrhosis or liver cancer, seriously threatening life and health.

[0007] Among them, alcoholic liver disease (ALD) is one of the typical representatives of liver injury. Long-term excessive drinking will lead to the accumulation of ethanol and its metabolite acetaldehyde in hepatocytes, causing mitochondrial dysfunction, oxidative stress, and inflammatory reactions, and finally progressing to steatosis (alcoholic fatty liver), hepatitis, and even liver fibrosis. Currently, the clinical treatment of ALD mainly focuses on abstinence from alcohol and drugs (such as metadoxine), but existing drugs have limitations:

[0008] 1. Side effects of metadoxine: As a central nervous system inhibitor, excessive use may cause drowsiness or coma;

[0009] 2. Metabolic burden: In patients with liver insufficiency or when combined with other medications, drug toxicity accumulation may occur due to decreased detoxification ability.

[0010] Therefore, it is urgent to develop a therapy with low side effects and effective in improving ALD. Plant extracts have become a research hotspot due to their advantages such as greenness, safety, and high biocompatibility, but existing preparations still have deficiencies:

[0011] 1. Single-component preparations (such as silymarin and glycyrrhizin): low bioavailability and single action targets;

[0012] 2. Compound preparation: Although it has the potential for multi-target regulation, there are bottlenecks in the preparation process:

[0013] 3. Water extraction method: It has low extraction efficiency for cell wall-bound components (such as gypenosides), and heat-sensitive components (such as polyphenols) are easily damaged at high temperatures;

[0014] 4. Solid-state fermentation: The metabolic capacity of a single strain is limited, and it is difficult to achieve synergistic effects of multiple components.

[0015] In summary, the existing liver protection preparations still face double dilemmas when dealing with alcoholic liver injury: chemical drugs have toxic side effects and metabolic burdens, while natural plant compound preparations are difficult to achieve efficient release and synergistic effects of active ingredients due to process limitations. Therefore, developing a new preparation process based on combined enzymatic hydrolysis and co-fermentation of multiple strains to break through the binding of plant cell wall structures to active ingredients and avoid the destruction of heat-sensitive components by high temperatures has become the key to improving the liver protection effect of plant compound preparations. This process can not only improve the yield and bioavailability of core functional components such as flavonoids and anthraquinones, but also synergistically relieve alcoholic liver injury through a multi-target regulation mechanism (antioxidation, anti-inflammatory, lipid metabolism regulation), providing a new technical path for the development of safe and efficient liver protection products. Summary of the Invention

[0016] The present invention provides a preparation method and use of an anti-alcoholic and liver-protecting composition, and the prepared anti-alcoholic and liver-protecting composition can effectively improve the liver function level.

[0017] To achieve the above object, the present invention adopts the following technical solutions:

[0018] In the first aspect, the present invention provides a preparation method of an anti-alcoholic and liver-protecting composition, comprising the following steps:

[0019] S1: Mix gynostemma powder, honeysuckle powder, hawthorn powder, green tea powder, lotus leaf powder, cassia seed powder, and senna leaf powder with sterile water to obtain a raw material liquid;

[0020] S2: Add cellulase to the raw material liquid and enzymatically hydrolyze for 3 - 5 h to obtain an enzymatically hydrolyzed liquid, wherein the pH of enzymatic hydrolysis is 5.0 - 5.5, and the temperature of enzymatic hydrolysis is 45 ± 5°C;

[0021] S3: Use sterile deionized water to prepare the enzymatically hydrolyzed liquid into a mixed liquid containing 11 - 13 wt%, inoculate the mixed liquid with a mixed strain liquid, and ferment for 18 - 22 h, wherein the fermentation temperature is 30 - 32°C, and the fermentation pH is 5 - 7;

[0022] S4: After the fermentation in S3 is completed, filter and sterilize the fermentation broth to obtain a fermentation filtrate, and concentrate and lyophilize the fermentation filtrate to obtain an anti-alcoholic and liver-protecting composition;

[0023] In step S1, the mass ratio of the gynostemma pentaphyllum powder, honeysuckle powder, hawthorn powder, green tea powder, lotus leaf powder, cassia seed powder, and senna leaf powder is 1:0.5 - 1:0.3 - 0.8:6 - 12:2 - 4:4 - 8:10 - 18.

[0024] In step S2, the addition amount of the cellulase is 0.3 - 0.5 wt% of the total mass of the gynostemma pentaphyllum powder, honeysuckle powder, hawthorn powder, green tea powder, lotus leaf powder, cassia seed powder, and senna leaf powder;

[0025] In step S3, the mixed bacterial strain solution is composed of a bran lactobacillus (preservation number: CGMCC No. 22055) bacterial strain solution and a salivary lactobacillus (preservation number: CCTCC M 2011127) bacterial strain solution;

[0026] The viable count in the bran lactobacillus bacterial strain solution is 1×10 10 CFU / g;

[0027] The viable count in the salivary lactobacillus bacterial strain solution is 1×10 10 CFU / g;

[0028] The mass ratio of the bran lactobacillus bacterial strain solution to the salivary lactobacillus bacterial strain solution in the mixed bacterial strain solution is 1:2 - 3;

[0029] The addition amount of the mixed bacterial strain is 5 - 8 wt% of the mixed solution.

[0030] In a second aspect, the present invention provides an anti - hangover and liver - protecting oral liquid, and the anti - hangover and liver - protecting oral liquid contains the following components in a mass ratio:

[0031] The anti - hangover and liver - protecting composition prepared by the preparation method described in the first aspect, honey, and drinking water, wherein the mass ratio of the anti - hangover and liver - protecting composition, honey, and drinking water is 1:0.5 - 0.7:1.3 - 1.8.

[0032] The present invention includes seven plant raw materials, namely, gynostemma pentaphyllum powder, honeysuckle flower powder, hawthorn powder, green tea powder, lotus leaf powder, cassia seed powder, and senna leaf powder. The above raw materials contain antioxidant substances such as flavonoids, phenols, tea polyphenols, and anthraquinones. These substances can jointly scavenge free radicals in the body, reduce the damage of oxidative stress to the liver, form an antioxidant synergistic effect, and effectively protect the integrity and functionality of hepatocytes. In addition, honeysuckle flower powder, green tea powder, cassia seed powder, and senna leaf powder all have anti-inflammatory properties. When used together, they can inhibit the inflammatory reaction in the liver, reduce the inflammatory damage of hepatocytes, and effectively relieve liver inflammation. Hawthorn powder, lotus leaf powder, and green tea powder can regulate blood lipid metabolism, reduce the levels of cholesterol and triglycerides in the blood, and reduce the accumulation of fat in the liver. At the same time, green tea powder can promote liver metabolism and enhance the liver's ability to metabolize and decompose various substances. The combined use of hawthorn powder, green tea powder, and lotus leaf powder helps to maintain the normal metabolic function of the liver and reduce the burden on the liver. Gynostemma pentaphyllum powder has a detoxifying effect and can help the liver decompose and remove toxins in the body. Cassia seed powder has a liver-protecting effect and can enhance the liver's detoxifying ability to protect hepatocytes from toxin damage. The two cooperate with each other and synergistically enhance the effect. Cassia seed powder and senna leaf can promote intestinal peristalsis, accelerate the excretion of toxins and waste in the body, reduce the absorption and reflux of toxins in the intestine, and indirectly protect the liver.

[0033] The Lactobacillus farraginis (preservation number: CGMCC No. 22055) and Lactobacillus salivarius (CCTCC M 2011127) used in the present invention are both fermentation strains that have been proven to be usable in foods. The present invention finds that by mixing and fermenting the two strains, more anthraquinones and flavonoids can be obtained than using a single strain under the same fermentation environment and strain dosage.

[0034] In addition to obtaining some beneficial secondary metabolites, the fermentation of the present invention can also dissolve the cell wall to release the substances inside the plant, which helps to improve the extraction efficiency.

[0035] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0036] (1) By using the enzymatic hydrolysis and fermentation extraction method, the present invention can retain as many different types of active and effective components as possible.

[0037] (2) Through fermentation, the present invention can increase the release of natural active substances, which can further enhance its liver-protecting effect.

[0038] (3) The solvents used throughout the extraction process of the present invention and the necessary substances used in the process are all non-toxic or low-toxic substances, and there is no residue after volatilization, ensuring the safety to the human body.

[0039] (4) The various active components in the composition finally obtained by the present invention synergistically enhance the effect. It has been proven by experiments that it has a statistically significant effect on liver protection. Brief Description of the Drawings

[0040] Figure 1 : Evaluation diagrams of the auxiliary protection effects of Compositions 1-3 and Compositions ①-⑨ against alcoholic liver injury.

[0041] Figure 2 : Effects of the mixed brewing solution of Gynostemma pentaphyllum powder, Lonicera japonica powder, Hawthorn powder, Green tea powder, Lotus leaf powder, Cassia seed powder, and Senna leaf powder on the physiological and biochemical indexes of alcohol-induced liver injury in mice. (A) Serum AST level, (B) Serum ALT level, (C) Liver index; all data are expressed as mean ± SEM (n = 6). a-d indicate significant differences in mice under different indexes, P < 0.05. Detailed Embodiments

[0042] To better understand the present invention, the present invention will be further described below in conjunction with specific embodiments. The terms used in the embodiments are for describing specific specific implementation manners and do not constitute a limitation on the protection scope of the present invention.

[0043] According to previous studies, the water extract prepared by hot water brewing and filtering using Gynostemma pentaphyllum powder, Lonicera japonica powder, Hawthorn powder, Green tea powder, Lotus leaf powder, Cassia seed powder, and Senna leaf powder as raw materials has a significant therapeutic effect on alcoholic liver injury. For details, see Figure 2 . The experimental period lasted for 57 days in total. Among them, the first week (days 1-7) was the animal adaptation period, and no treatment was applied to the mice in each group. The second week (days 8-14) was the pretreatment stage: The mice in the normal group (Control) and the model group (EtOH) were gavaged with distilled water once a day; The mice in the low-dose group (EtOH+L), the middle-dose group (EtOH+M), and the high-dose (EtOH+H) group were gavaged with the corresponding dose of the above-mentioned tea water extract once a day (low dose 0.65 g / kg, middle dose 1.30 g / kg, high dose 2.60 g / kg). The third to eighth weeks (days 15-57) were the alcoholic liver injury model induction and drug intervention stage, in which: The mice in the normal group were gavaged with distilled water twice a day, and the interval between the two gavaging treatments was 3 h; The mice in the model group were first gavaged with distilled water once a day, and 3 h later, they were continuously gavaged with 30% (V / V) ethanol solution for 14 days, then continuously gavaged with 40% (V / V) ethanol solution for 14 days, and finally continuously gavaged with 50% (V / V) ethanol solution for 14 days, once a day, and the dose each time was 0.1 mL / 10 g body weight; The mice in the low-dose group, the middle-dose group, and the high-dose group were first gavaged with the corresponding dose of the tea water extract once a day, and 3 h later, they were continuously gavaged with 30% (V / V) ethanol solution for 14 days, 40% (V / V) ethanol solution for 14 days, and 50% (V / V) ethanol solution for 14 days, once a day, and the dose each time was 0.1 mL / 10 g body weight.

[0044] Figure 2 The results of A showed that compared with the normal group, the serum AST level of mice in the model group was significantly increased, indicating that the alcoholic liver injury model was successfully established. After intragastric administration of tea water at low dose (0.65 g / kg), medium dose (1.30 g / kg) and high dose (2.60 g / kg), the serum AST levels of mice in each dose group were significantly lower than those in the model group, and there was no statistical difference among the low, medium and high dose treatments (P>0.05). Figure 2 The results of B showed that the serum ALT level of mice in the model group was significantly increased (20.17% higher than that in the normal group). There was no significant difference in the serum ALT level between the low dose group and the model group, while the ALT levels of mice in the medium dose group and the high dose group were significantly decreased to the level equivalent to that in the normal group. Therefore, based on the detection results of AST and ALT, it can be known that the alleviating effect of intragastric administration of medium dose and high dose tea water on alcohol-induced liver injury is significantly better than that of the low dose group. Figure 2 The results of C further showed that the liver index of mice in the model group was significantly increased (18.84% higher than that in the normal group). Among the dose groups given tea water intervention, only the liver index of the high dose group was significantly decreased compared with that in the model group (decreased by 10.02%), and recovered to the level without statistical difference from that in the normal group. This shows that the alleviating effect of intragastric administration of high dose tea water on alcohol-induced liver injury is significantly better than that of the medium and low dose groups, and the intervention effect is the best. Based on the above results, the present invention further explores taking gynostemma pentaphyllum powder, honeysuckle powder, hawthorn powder, green tea powder, lotus leaf powder, cassia seed powder, senna leaf powder as raw materials and further extracting through a fermentation process in order to further enhance its liver protection effect.

[0045] The raw materials and reagents described in the present invention are all commercially available.

[0046] The water described in the present invention is drinking water unless otherwise specified.

[0047] Example 1: Preparation of an anti-alcohol and liver-protecting composition

[0048] An anti-alcohol and liver-protecting composition, and its preparation method is as follows:

[0049] Composition 1:

[0050] Step 1: Mixing: Weigh each raw material according to the mass ratio of 1:0.7:0.5:9:3:6:14 for gynostemma pentaphyllum powder, honeysuckle powder, hawthorn powder, green tea powder, lotus leaf powder, cassia seed powder, senna leaf powder, mix them to obtain a raw material powder, and mix the raw material powder with sterile water at a mass ratio of 1:12 to obtain a raw material liquid.

[0051] Step 2: Enzymatic hydrolysis: Add cellulase to the raw material liquid, and perform enzymatic hydrolysis at 45 °C for 4 h to obtain an enzymatic hydrolysate. The addition amount of cellulase is 0.4 wt% of the total mass of gynostemma powder, honeysuckle powder, hawthorn powder, green tea powder, lotus leaf powder, cassia seed powder, and senna leaf powder.

[0052] Step 3: Fermentation: Use sterile deionized water to prepare a mixed solution containing 12 wt% of the enzymatic hydrolysate. After sterilizing the mixed solution, inoculate a mixed strain solution accounting for 6 wt% of the mixed solution, and ferment at 31 °C for 20 h. The initial pH of fermentation is 6. After fermentation, a fermented turbid liquid is obtained. The mixed strain solution is composed of Lactobacillus farraginis CGMCC No. 22055 strain solution and Lactobacillus salivarius CCTCC M 2011127 strain solution mixed in a mass ratio of 1:2.5.

[0053] Step 4: Ultrasonic lysis: Perform ultrasonic lysis on the fermented turbid liquid, then filter and irradiate for sterilization to obtain a fermented filtrate, where the ultrasonic frequency is 30 kHz and the ultrasonic time is 45 min.

[0054] Step 5: Freeze-drying: Concentrate the fermented filtrate to 30% of the original volume, and obtain a liver-protecting composition through low-temperature freeze-drying technology.

[0055] The viable count of the Lactobacillus farraginis CGMCC No. 22055 strain solution is: 1×10 10 CFU / g;

[0056] The viable count of the Lactobacillus salivarius CCTCC M 2011127 strain solution is: 1×10 10 CFU / g.

[0057] For Composition 2-3, the steps are the same as those of Composition 1, and the specific technical parameters are shown in Table 1 below. The technical parameters not involved in Table 1 are the same as those of Composition 1.

[0058] Table 1 Technical Parameters

[0059]

[0060] Composition ①: Different from Composition 1, Saccharomyces cerevisiae is used to replace Lactobacillus farraginis CGMCC No. 22055, and the other conditions and parameters are the same as those of Composition 1. The Saccharomyces cerevisiae is purchased from Angel Yeast Co., Ltd.

[0061] Composition ②: Different from Composition 1, commercial Lactobacillus salivarius is used to replace Lactobacillus salivarius CCTCC M 2011127, and the other conditions and parameters are the same as those of Composition 1. The commercial Lactobacillus salivarius is purchased from Guangzhou Huijian Biotechnology Co., Ltd.

[0062] Composition ③: Different from Composition 1, the mass ratio of Lactobacillus farraginis to Lactobacillus salivarius is 2.5:1, and the remaining conditions and parameters are the same as those of Composition 1.

[0063] Composition ④: Different from Composition 1, the mass ratio of Lactobacillus farraginis to Lactobacillus salivarius is 1:1, and the remaining conditions and parameters are the same as those of Composition 1.

[0064] Composition ⑤: Different from Composition 1, the raw material used lacks green tea powder, and the lacking mass parts are supplemented with gynostemma pentaphyllum powder, honeysuckle powder, hawthorn powder, lotus leaf powder, cassia seed powder, and senna leaf powder with a mass ratio of 1:0.7:0.5:3:6:14, and the remaining conditions and parameters are the same as those of Composition 1.

[0065] Composition ⑥: Different from Composition 1, the raw material used lacks cassia seed powder, and the lacking mass parts are supplemented with gynostemma pentaphyllum powder, honeysuckle powder, hawthorn powder, green tea powder, lotus leaf powder, and senna leaf powder with a mass ratio of 1:0.7:0.5:9:3:14, and the remaining conditions and parameters are the same as those of Composition 1.

[0066] Composition ⑦: Different from Composition 1, the raw material used lacks senna leaf powder, and the lacking mass parts are supplemented with gynostemma pentaphyllum powder, honeysuckle powder, hawthorn powder, green tea powder, lotus leaf powder, and cassia seed powder with a mass ratio of 1:0.7:0.5:9:3:6, and the remaining conditions and parameters are the same as those of Composition 1.

[0067] Composition ⑧: Different from Composition 1, the mass ratio of gynostemma pentaphyllum powder, honeysuckle powder, hawthorn powder, green tea powder, lotus leaf powder, cassia seed powder, and senna leaf powder in the raw material used is 0.5:1:0.7:14:3:9:6, and the remaining conditions and parameters are the same as those of Composition 1.

[0068] Composition ⑨: Different from Composition 1, step 3 is not performed, and the enzyme hydrolyzate is ultrasonically lysed and freeze-dried, and the remaining conditions and parameters are the same as those of Composition 1.

[0069] Example 2: Determine the total anthraquinone and total flavonoid contents in Compositions 1-3, Compositions ①-④, and ⑨

[0070] The test method for the total anthraquinone content refers to "Determination of Total Anthraquinones in Health Foods" (Ye Bisha et al., Chinese Journal of Health Inspection, DOI: 10.3969 / j.issn.1004-8685.2007.05.027).

[0071] The determination method for the total flavonoid content is carried out according to the method described in "DB 34 / T 2743-2016 Determination of Total Flavonoid Content in Sophora japonica and Its Products - Spectrophotometry".

[0072] Table 2 Determination results of total anthraquinone and total flavonoid contents

[0073]

[0074] The data in Table 2 show that for the composition prepared by the specific process of the present invention, the contents of total flavonoids and total anthraquinones have both increased.

[0075] Example 3: Animal toxicological safety test

[0076] 1) Materials and methods

[0077] 1. Test substances: Samples of Compositions 1 to 3; Prepared into a 1 g / mL mixture using drinking water.

[0078] 2. Acute toxicity experiment: Sixty Kunming mice with a body weight of 20 ± 2 g, half male and half female, were selected; Environment: Temperature 23 ± 1°C, humidity 54 ± 2%.

[0079] The concentrated solutions of Samples of Compositions 1 - 3 were orally administered to mice by gavage once, with a gavage volume of 0.2 mL / 10 g·b.w., corresponding to a dose of 20.0 g / kg·b.w. The mice were fasted for 16 hours before gavage and continuously observed for two weeks after gavage, and the toxic manifestations and death conditions were recorded, as shown in Table 3.

[0080] Table 3 Acute toxicity test of mice

[0081] Group Gender Route Dose (g / kg·b.w.) Number of deaths (animals) MTD (g / kg·b.w.) Composition 1 Male Oral 20.0 0 >20.0 Composition 1 Female Oral 20.0 0 >20.0 Composition 2 Male Oral 20.0 0 >20.0 Composition 2 Female Oral 20.0 0 >20.0 Composition 3 Male Oral 20.0 0 >20.0 Composition 3 Female Oral 20.0 0 >20.0

[0082] According to the results in Table 3, no mice died in this experiment, indicating that the composition provided in this test has high biological safety.

[0083] Example 4: Animal efficacy test

[0084] Preparation of test sample solutions:

[0085] Test group: Compositions 1 - 3, Compositions ① - ⑨, with deionized water as the solvent;

[0086] Normal control group: Deionized water.

[0087] Positive control group: Metadoxine capsules (hereinafter referred to as metadoxine), white powder, batch number 230801, Zhejiang Zhenyuan Pharmaceutical Co., Ltd., with deionized water as the solvent.

[0088] Experimental animals:

[0089] Zebrafish were all raised in fish culture water at 28°C (Water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, conductivity was 500 μS / cm; pH was 7.0; hardness was 80 mg / L CaCO3).

[0090] Instruments, consumables and reagents:

[0091] Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); Precision electronic balance (CP214, OHAUS, USA); Ultrasonic cleaner (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China); 6-well plate (Zhejiang Beilanbo Biotechnology Co., Ltd., China).

[0092] Absolute ethanol (batch number F2209028, Shanghai Aladdin Biochemical Technology Co., Ltd., China).

[0093] Detection method:

[0094] Wild-type AB strain zebrafish at 3 days post-fertilization (3dpf) were randomly selected and placed in a 6-well plate, with 30 zebrafish in each well. The samples were administered in water (concentrations are shown in Table 4). At the same time, a normal control group and a model control group were set up, and the volume of each well was 3 mL. Except for the normal control group, the remaining experimental groups were administered absolute ethanol in water, so that the final concentration of ethanol in the environment was 2 v / v%, to establish an alcoholic liver injury model. After 2 days of treatment, the MTC of the samples on the model zebrafish was measured. During the experiment, the temperature of the environmental water body was 28 ± 1°C.

[0095] Evaluation of the efficacy of relieving hangover and protecting the liver:

[0096] Wild-type AB strain zebrafish at 3dpf were randomly selected and placed in a 6-well plate, with 30 zebrafish in each well. The samples were administered in water (concentrations are shown in Table 5). The concentration of the positive control group, metadoxine, was 149 μg / mL. At the same time, a normal control group and a model control group were set up. The normal control group and the model control group were not given the samples, and the volume of each well was 3 mL. Except for the normal control group, the remaining experimental groups were administered absolute ethanol in water, so that the ethanol concentration in the breeding environment reached 2 v / v%, to establish an alcoholic liver injury model. After 2 days of treatment in an environmental water body at 28 ± 1°C, 10 zebrafish were randomly selected from each experimental group and photographed under a dissecting microscope. The NIS-Elements D 3.20 advanced image processing software was used to analyze and collect data, and the liver area, average liver brightness, and delayed yolk sac absorption area of the zebrafish were analyzed. The auxiliary protective effect of the samples against alcoholic liver injury was evaluated based on the statistical analysis results of the above indicators. The statistical processing results were expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0097] Results:

[0098] MTC (Minimum toxic concentration):

[0099] Under the experimental conditions, the MTC of Composition 1, Composition 2 and Composition 3 > 2000 μg / mL. See Table 4 for details.

[0100] Table 4 MTC Detection

[0101]

[0102] Evaluation of the effect of relieving hangover and protecting the liver:

[0103] Under the experimental conditions, Composition 1, Composition 2 and Composition 3 have the effect of relieving hangover and protecting the liver. See Table 5 for details.

[0104] Table 5 Evaluation of liver protection effect

[0105]

[0106]

[0107] Note: Compared with the model control group, *p < 0.05, **p < 0.01, ***p < 0.001

[0108] According to the results in Table 5, compared with the model control group, Compositions 1-3 all have a significant liver protection effect.

[0109] By comparing Compositions ①-⑨ with Composition 1, it can be seen that at a dose of 1000 μg / mL, Composition 1 has the best effect.

[0110] By comparing Composition 1 with Compositions ①-②, it can be seen that there is a significant synergistic effect between Lactobacillus farraginis CGMCC NO.26508 and Lactobacillus salivarius CCTCC M 2011127 selected in the present invention, which can improve the liver protection effect of the composition.

[0111] By comparing Composition 1 with Compositions ③-④, it can be seen that the mass ratio of Lactobacillus farraginis to Lactobacillus salivarius selected in the present invention has a significant synergistic effect within the range defined in the present invention, which can improve the liver protection effect of the composition.

[0112] By comparing Composition 1 with Compositions ⑤-⑧, it can be seen that the fermentation raw materials of the present invention have a certain synergistic effect, and within the range of the raw material types and their mass fraction ratios defined in the present invention, the obtained composition has the best liver protection effect.

[0113] By comparing Composition 1 with Composition ⑨, it can be seen that the selected bacterial agent and fermentation method in the present invention have a certain promoting effect on the manifestation of the liver protection effect of the composition of the present invention.

[0114] Application example: Components and preparation method of hangover and liver protection oral liquid:

[0115] Hangover and liver protection oral liquid 1:

[0116] Add 15 g of water to 10 g of the hangover-relieving and liver-protecting composition (Composition 1), homogenize for 4 min, then add 6 g of honey and homogenize for 6 min to obtain Hangover-Relieving and Liver-Protecting Oral Liquid 1.

[0117] Hangover-Relieving and Liver-Protecting Oral Liquid 2:

[0118] Add 13 g of water to 10 g of the hangover-relieving and liver-protecting composition (Composition 1), homogenize for 3 min, then add 5 g of honey and homogenize for 8 min to obtain Hangover-Relieving and Liver-Protecting Oral Liquid 2.

[0119] Hangover-Relieving and Liver-Protecting Oral Liquid 3:

[0120] Add 18 g of water to 10 g of the hangover-relieving and liver-protecting composition (Composition 1), homogenize for 5 min, then add 7 g of honey and homogenize for 5 min to obtain Hangover-Relieving and Liver-Protecting Oral Liquid 3.

[0121] The above detailed description is a specific description of one of the feasible embodiments of the present invention. This embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or change that does not depart from the present invention should be included within the scope of the technical solution of the present invention.

Claims

1. A preparation method of an anti-hangover and liver-protecting composition, characterized in that, The method comprises the following steps: S1: Mix gynostemma pentaphyllum powder, honeysuckle flower powder, hawthorn powder, green tea powder, lotus leaf powder, cassia seed powder, and senna leaf powder with sterile water to obtain a raw material liquid; S2: Add cellulase to the raw material liquid and enzymatically hydrolyze for 3 - 5 h to obtain an enzymatically hydrolyzed liquid, wherein the pH of enzymatic hydrolysis is 5.0 - 5.5, and the temperature of enzymatic hydrolysis is 45 ± 5°C; S3: Use sterile deionized water to prepare the enzymatically hydrolyzed liquid into a mixed liquid containing 11 - 13 wt%, inoculate the mixed liquid with a mixed strain liquid, and ferment for 18 - 22 h, wherein the fermentation temperature is 30 - 32°C, and the initial pH of fermentation is 5 - 7; S4: After the fermentation in S3 ends, filter and sterilize the fermentation broth to obtain a fermented filtrate, and concentrate and freeze-dry the fermented filtrate to obtain an anti-alcoholic and liver-protecting composition; Among them, in step S1, the mass ratio of the gynostemma pentaphyllum powder, honeysuckle flower powder, hawthorn powder, green tea powder, lotus leaf powder, cassia seed powder, and senna leaf powder is 1:0.5 - 1:0.3 - 0.8:6 - 12:2 - 4:4 - 8:10 - 18; In step S2, the addition amount of the cellulase is 0.3 - 0.5 wt% of the total mass of the gynostemma pentaphyllum powder, honeysuckle flower powder, hawthorn powder, green tea powder, lotus leaf powder, cassia seed powder, and senna leaf powder; The mixed bacterial strain solution described in step S3 is composed of a strain solution of Lactobacillus farinorum with a preservation number of CGMCC No. 22055 and a strain solution of Lactobacillus salivarius with a preservation number of CCTCC M 2011127; the mass ratio of the Lactobacillus farinorum strain solution to the Lactobacillus salivarius strain solution is 1:2-3; the addition amount of the mixed bacterial solution is 5-8 wt% of the mixed solution; the viable count of the Lactobacillus farinorum strain solution is 1×10 8 -1×10 10 CFU / g; the viable count of the Lactobacillus salivarius strain solution is 1×10 8 -1×10 10 CFU / g.

2. The preparation method of the hangover and liver-protecting composition according to claim 1, characterized in that, In step S1, the mass ratio of the total mass of the gynostemma pentaphyllum powder, honeysuckle flower powder, hawthorn powder, green tea powder, lotus leaf powder, cassia seed powder, and senna leaf powder to the mass of sterile water is 1:11 - 13.

3. The preparation method of the hangover and liver-protecting composition according to claim 1 or 3, characterized in that, The mass ratio of the Lactobacillus farraginis to the Lactobacillus salivarius is 1:2.

5.

4. A liver-protecting product, characterized in that, The liver-protecting product contains the anti-alcoholic and liver-protecting composition prepared by the preparation method according to any one of claims 1 - 3, and the dosage form of the liver-protecting product is selected from any one of ointment, tablet, capsule, granule, or oral liquid.

5. A preparation method of an oral liquid for relieving hangover and protecting the liver, characterized in that, The method comprises the following steps: Add drinking water to the anti-alcoholic and liver-protecting composition according to claim 1, homogenize for 3 - 5 min, and then add honey and homogenize for 5 - 8 min to obtain an anti-alcoholic and liver-protecting oral liquid.

6. The preparation method of the hangover and liver-protecting oral liquid according to claim 5, wherein, The mass ratio of the anti-alcoholic and liver-protecting composition, honey, and drinking water is 1:0.5 - 0.7:1.3 - 1.8.

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