A probiotic fermented liver-protecting and alcohol-dissipating flower tea composition, a preparation method and application thereof

Abstract

This invention discloses a probiotic-fermented herbal tea composition for liver protection and hangover relief, its preparation method, and its application. The composition comprises roselle, dandelion, kudzu, Panax notoginseng, and Dendrobium officinale flowers. The extract is obtained through ultrasonic extraction with ethanol, followed by fermentation with Lactobacillus plantarum, and then freeze-dried to obtain a freeze-dried powder. This herbal tea composition has liver-protecting and hangover-relieving effects and can be used to prepare drugs, health products, or foods for preventing and treating alcoholic liver damage and alleviating hangovers.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine compound preparation technology, specifically relating to a probiotic fermented herbal tea composition for liver protection and hangover relief, its preparation method and application. Background Technology

[0002] With the fast pace of modern society and the increased frequency of social engagements, drinking alcohol has become an unavoidable part of daily life for most people. However, liver damage caused by long-term or excessive drinking is becoming increasingly prominent, especially acute alcoholic liver injury. Modern medical research shows that alcoholic liver injury is a liver disease induced by long-term heavy drinking: ethanol and its metabolites have direct toxicity to the liver, while the liver's metabolic and detoxification capacity decreases, leading to pathological changes such as hepatocyte steatosis, inflammation, and fibrosis, which can develop into cirrhosis or even liver cancer in severe cases. Currently, commonly used hepatoprotective drugs such as silymarin have a certain protective effect on hepatocytes, but their effects on delaying intoxication and accelerating sobering up are not significant, and most of them are chemical monomeric preparations, raising concerns about side effects with long-term use.

[0003] From the perspective of Traditional Chinese Medicine (TCM), alcohol (anciently known as "alcohol poison") is pungent and hot in nature. Excessive consumption can lead to stagnation of liver qi, internal generation of damp-heat, and depletion of body fluids. Since the liver governs the free flow of qi, alcohol obstructs this flow, causing stagnation and symptoms such as headache and distension in the hypochondrium. Simultaneously, alcohol poison easily generates damp-heat, damaging the spleen and stomach, leading to nausea and vomiting, and depleting body fluids, resulting in dry mouth and dizziness. TCM has accumulated thousands of years of rich experience in treating liver and stomach diseases, adhering to the core principles of holistic conditioning and syndrome differentiation, demonstrating unique advantages. The clinical efficacy of traditional Chinese medicine formulas passed down through generations, classic prescriptions, and modern optimized formulas for liver and stomach-related diseases generally exceeds 90%. Furthermore, these formulas are mild in nature, focus on maintaining bodily balance, and have far fewer side effects than chemical preparations, providing excellent research and development ideas for the prevention and treatment of alcoholic liver and stomach damage. Based on ancient texts and modern research, Compound Fresh Dendrobium Granules is a mature traditional Chinese medicine formula that can be developed into a health food. Its ingredients are formulated with fresh Dendrobium, kudzu root, and Panax notoginseng in a ratio of 6:8:1. It has the core effects of nourishing yin and stomach, clearing heat and relieving alcohol, and promoting body fluid and quenching thirst. It can not only relieve the irritation of alcohol on the gastric mucosa, but also provide initial protection for the liver, laying the foundation for the subsequent development of natural liver-protecting and hangover-relieving products.

[0004] In recent years, the application of probiotic fermentation technology in traditional Chinese medicine compound prescriptions has received increasing attention. Studies have shown that probiotics such as *Lactobacillus plantarum* possess good safety and fermentation adaptability, and their metabolic activities can alter the release and transformation of substrate active ingredients. However, there are currently no research reports on applying *Lactobacillus plantarum* fermentation technology to specific ratio herbal tea compositions to synergistically enhance liver-protecting and hangover-relieving effects. Furthermore, low-temperature vacuum freeze-drying technology can lock in the active nutrients of raw materials to the greatest extent, effectively extending the product's shelf life. The finished product also boasts excellent solubility and convenient consumption, better meeting modern consumers' demands for natural, efficient, and portable health foods.

[0005] However, existing liver-protecting and hangover-relieving products generally have the following shortcomings: First, the retention rate of active ingredients is low, and the dissolution rate of effective ingredients in traditional tea bags or decoctions is limited; second, it is difficult to achieve both hangover relief and liver protection effects, with most products focusing only on one aspect; and third, there is a lack of mature technology to enhance the efficacy of flower-based medicinal materials through probiotic fermentation.

[0006] While ancient texts record the use of floral medicinal herbs such as hibiscus, kudzu, and Panax notoginseng for relieving hangovers, these are mostly used alone or in simple combinations. Systematic research on the synergistic relationships between these herbs is lacking, particularly in the technical solutions for combining specific herbal tea compositions with probiotic fermentation processes to achieve synergistic effects. Therefore, developing a product with scientifically formulated components, offering both hangover relief and liver protection, preserving active ingredients, and convenient for carrying and consumption has significant practical value. Summary of the Invention

[0007] The purpose of this invention is to overcome the shortcomings of the prior art and provide a flower tea composition and its preparation method that has sufficient retention of active ingredients, significant liver protection and hangover relief effects, and is natural and safe.

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] A probiotic fermented flower tea composition for liver protection and hangover relief, wherein the composition is obtained by fermenting a flower tea extract with probiotics, the flower tea extract being composed of the following flower teas in parts by weight: 10-20 parts roselle, 8-12 parts dandelion, 8-12 parts kudzu, 2-4 parts Panax notoginseng, and 7-11 parts Dendrobium officinale; the probiotic is Lactobacillus plantarum.

[0010] Furthermore, the liver-protecting and hangover-relieving probiotic fermented flower tea composition is prepared by fermenting flower tea extract with probiotics; the flower tea extract is composed of the following flower teas in parts by weight: 15 parts roselle, 10 parts dandelion, 10 parts kudzu, 3 parts Panax notoginseng, and 9 parts Dendrobium officinale; the probiotic is Lactobacillus plantarum.

[0011] Furthermore, in the probiotic fermented flower tea composition, the effective live bacteria count of the probiotics is 1.0 × 10⁻⁶. 7 cfu / mL~1.0×10 9 cfu / mL, preferably 1.0 × 10⁻⁶ 9 cfu / mL.

[0012] Furthermore, a freeze-dried powder is prepared by freeze-drying the aforementioned probiotic fermented herbal tea composition.

[0013] Furthermore, the freeze-dried powder is prepared by the following steps:

[0014] Step (1) Preparation of flower tea extract: Mix the raw materials of flower tea evenly according to the ratio, add 60% ethanol at a solid-liquid ratio of 1:40 (g / mL), and extract under biological conditions of 70℃, ultrasonic frequency of 40kHZ and power of 100W for 50min to obtain the extract; concentrate the extract under reduced pressure until there is no alcohol taste to obtain flower tea medicinal liquid.

[0015] Step (2) Preparation of probiotic culture: Lactobacillus plantarum was inoculated onto MRS solid medium and cultured under anaerobic conditions at 37℃ for screening and purification; single colonies were picked and inoculated into MRS liquid medium and cultured under anaerobic conditions at 37℃ to obtain Lactobacillus plantarum culture; the effective viable count of the culture was adjusted to 1.0 × 10⁻⁶. 9 cfu / mL;

[0016] Step (3) Preparation of probiotic fermented flower tea liquid: The probiotic liquid obtained in step (2) is inoculated into the flower tea liquid at 5% of the volume of the flower tea liquid obtained in step (1), and fermented under anaerobic conditions at 37℃ for 72 hours to obtain probiotic fermented flower tea liquid.

[0017] Step (4) Preparation of freeze-dried powder: The probiotic fermented flower tea liquid obtained in step (3) is centrifuged at 10000 r / min for 20 min, filtered through a 0.22 μm microporous membrane to remove impurities, and the filtrate is collected; the filtrate is pre-frozen at -45℃ for 4 h, and then freeze-dried at a vacuum of 20 Pa and a sublimation temperature of -15℃ until the moisture content does not exceed 5%, then ground into powder and passed through a 100-mesh sieve to obtain freeze-dried powder.

[0018] Furthermore, the use of the probiotic fermented flower tea composition or the freeze-dried powder of claim 4 in the preparation of products with liver-protecting and / or hangover-relieving effects.

[0019] Furthermore, in the aforementioned applications, the product is a pharmaceutical, health product, or food.

[0020] Furthermore, in the aforementioned application, the dosage of the probiotic fermented flower tea composition or lyophilized powder in the mouse model of the product is 0.76 g / kg based on the lyophilized powder, which is equivalent to 3.8 g / kg of raw medicinal material.

[0021] Furthermore, in the aforementioned application, the liver-protective effects include reducing serum alanine aminotransferase (ALT) and / or aspartate aminotransferase (AST) levels in individuals with alcoholic liver injury, and / or increasing liver superoxide dismutase (SOD) activity and / or glutathione (GSH) content, and / or reducing liver malondialdehyde (MDA) content; the hangover relief effects include prolonging the latency period of intoxication and / or increasing the success rate of sobering up and / or shortening the sobering up time.

[0022] Alcohol is a pungent, sweet, and extremely hot substance, characterized by its potent and slippery nature, and its toxicity. From the perspective of traditional Chinese medicine, alcohol poisoning is essentially a pathological process in which the pungent and hot toxic pathogens of alcohol invade the body, spreading along the meridians and damaging the internal organs. Initially, alcohol poisoning directly attacks the spleen and stomach, its pungent and hot nature scorching the stomach and causing disharmony in the stomach's descending function and impaired digestion. Its damp-heat nature also obstructs the spleen, causing it to lose its ability to transform and transport nutrients, and preventing the clear yang from ascending. The spleen is the pivot of qi's ascending and descending function and the core of water and dampness transportation. When damaged, water and dampness stagnate, combining with the heat and toxicity of alcohol to form a syndrome of damp-heat trapping the spleen, manifesting as initial symptoms after drinking such as nausea and vomiting, abdominal distension, loss of appetite, bitter and sticky taste in the mouth, and heaviness in the limbs. If damp-heat in the spleen and stomach persists for a long time without being resolved, the pathogenic factors will spread from the viscera to the internal organs, entering the liver and gallbladder. The liver and gallbladder are responsible for regulating the flow of qi and assisting the spleen and stomach in their digestive functions. When damp-heat accumulates in the liver and gallbladder, it causes the liver to lose its ability to regulate qi, leading to stagnation of qi and the transformation of stagnation into fire, resulting in symptoms of damp-heat in the liver and gallbladder. Symptoms include distending pain in the hypochondrium, irritability, red eyes, dry mouth, and dark yellow urine. The liver is already damaged, and alcoholic liver damage will subsequently appear. Therefore, the treatment of alcoholic liver and stomach damage should follow the basic principles of clearing away alcohol toxins, protecting the spleen and stomach, soothing the liver and gallbladder, nourishing yin and generating fluids, and addressing both the symptoms and the root cause.

[0023] Based on the above understanding of the pathogenesis, this invention selects five floral materials—roselle, dandelion, kudzu, Panax notoginseng, and Dendrobium officinale—to formulate a prescription that closely follows the pathogenesis of alcohol poisoning. Each material performs its function and works synergistically to achieve the effect of protecting the liver and relieving alcohol intoxication. The formula uses roselle and dandelion as the principal herbs: Roselle is cold in nature and sour in taste, entering the liver meridian. It is light and soft in texture. Following the principle that "sourness can astringe and sweetness can nourish yin", it can clear heat and cool blood, generate fluids and relieve irritability. It can clear the liver and stomach heat caused by alcohol poisoning and protect the liver by cooling blood and reducing the direct damage of alcohol poisoning to the liver. Dandelion is cold in nature and bitter in taste, entering the liver and stomach meridians. It is good at clearing heat and detoxifying, clearing damp heat. It works synergistically with roselle to clear the body of alcohol poisoning and metabolic turbidity. The combination of sour and bitter herbs is in line with the physiological characteristics of the liver that is "yin in substance and yang in function". It can clear the liver fire caused by alcohol poisoning and nourish the yin fluid of the liver meridian, clearing without damaging the liver yin. Kudzu flower, used as an assistant herb, is cool in nature and slightly sweet in taste. It enters the spleen and stomach meridians and is effective in relieving alcohol poisoning, invigorating the spleen, clearing heat and detoxifying, and nourishing qi and yin. It can resolve the evil qi of alcohol, reduce the damage of alcohol to the spleen and stomach's digestive function, invigorate the spleen and stomach, and improve discomforts such as nausea, abdominal distension, and poor appetite after drinking. At the same time, its cool nature helps the principal herb to clear accumulated heat in the liver and stomach, and prevent the transmission of damp-heat pathogens from the spleen and stomach to the liver and gallbladder. Panax notoginseng flower, used as an adjuvant herb, is cool in nature and slightly bitter in taste. It enters the liver and heart meridians and has the effects of promoting blood circulation, removing blood stasis, and soothing the liver and regulating qi. Prolonged alcohol poisoning can easily lead to qi stagnation and meridian obstruction, which can cause discomfort in the hypochondrium, chest tightness, and irritability. This herb can soothe the liver, unblock the meridians, remove blood stasis, and relieve various symptoms of qi stagnation and blood stasis after drinking. Dendrobium officinale flower is used as the guiding herb. It is slightly cold in nature and sweet in taste. It enters the stomach and kidney meridians and is good at nourishing yin and generating fluids, benefiting the stomach and protecting the liver, and harmonizing other herbs. Alcohol is pungent and hot and can easily deplete the yin fluids of the liver and stomach. This herb can nourish the yin fluids of the liver and stomach and replenish the yin deficiency caused by alcohol poisoning. At the same time, it can harmonize the properties and flavors of the whole formula and buffer the cold and cooling properties of other herbs, so that the whole formula is clear without being cold, draining without being harmful, and nourishing without being stagnant.

[0024] In terms of process, this invention introduces a fermentation process using *Lactobacillus plantarum*, resulting in the following synergistic effects: the enzymes produced by probiotic metabolism can degrade macromolecules in the flowers, promoting the conversion of flavonoids into aglycones with higher bioavailability and improving the absorption rate of active ingredients; the short-chain fatty acids and other metabolites produced during fermentation have anti-inflammatory and intestinal barrier protective functions, complementing the antioxidant effects of the flower tea components; simultaneously, probiotics can regulate the intestinal microecology, providing internal environmental support for the positive regulation of the gut-liver axis. Combined with freeze-drying technology, the active ingredients of the fermentation products are preserved to the greatest extent, extending the product's shelf life. Attached Figure Description

[0025] Figure 1 Effects of herbal teas for liver protection and hangover relief on the latency period of acute alcohol intoxication in mice;

[0026] Figure 2 The effect of herbal teas for liver protection and hangover relief on the sobering time of acutely intoxicated mice;

[0027] Figure 3Effects of herbal teas for liver protection and hangover relief on serum ALT levels in acutely intoxicated mice;

[0028] Figure 4 Effects of herbal teas for liver protection and hangover relief on serum AST levels in acutely intoxicated mice;

[0029] Figure 5 Effects of herbal teas for liver protection and hangover relief on SOD levels in the livers of acutely intoxicated mice;

[0030] Figure 6 Effects of herbal teas for liver protection and hangover relief on liver MDA levels in acutely intoxicated mice;

[0031] Figure 7 The effect of herbal teas for liver protection and hangover relief on GSH levels in the livers of acutely intoxicated mice;

[0032] Figure 8 The effects of herbal teas for liver protection and hangover relief on the histopathology of mouse liver tissue. Detailed Implementation

[0033] The specific embodiments of the present invention will be described in detail below with reference to examples.

[0034] Roselle, dandelion, kudzu, Panax notoginseng, and Dendrobium officinale flowers were purchased from the Bozhou Traditional Chinese Medicine Market and meet the relevant standards of the 2025 edition of the Pharmacopoeia of the People's Republic of China. Lactobacillus plantarum, accession number CGMCC15013 (published in CN109423467A).

[0035] Example 1:

[0036] Formula ratio: 150g roselle, 100g dandelion, 100g kudzu, 30g Panax notoginseng, 90g Dendrobium officinale, and Lactobacillus plantarum inoculated at 5% of the tea liquid volume, with a bacterial concentration of 1.0 × 10⁻⁶. 9 cfu / mL.

[0037] The preparation method of the embodiment is as follows:

[0038] Step (1) Preparation of flower tea extract: Mix the raw materials of flower tea evenly according to the ratio, add 60% ethanol at a solid-liquid ratio of 1:40 (g / mL), and extract under biological conditions of 70℃, ultrasonic frequency of 40kHZ and power of 100W for 50min to obtain the extract; concentrate the extract under reduced pressure until there is no alcohol taste to obtain flower tea medicinal liquid.

[0039] Step (2) Preparation of probiotic culture: Lactobacillus plantarum was inoculated onto MRS solid medium and cultured under anaerobic conditions at 37℃ for screening and purification; single colonies were picked and inoculated into MRS liquid medium and cultured under anaerobic conditions at 37℃ to obtain Lactobacillus plantarum culture; the effective viable count of the culture was adjusted to 1.0 × 10⁻⁶. 9 cfu / mL;

[0040] Step (3) Preparation of probiotic fermented flower tea liquid: The probiotic liquid obtained in step (2) is inoculated into the flower tea liquid at 5% of the volume of the flower tea liquid obtained in step (1), and fermented under anaerobic conditions at 37℃ for 72 hours to obtain probiotic fermented flower tea liquid.

[0041] Step (4) Preparation of freeze-dried powder: The probiotic fermented flower tea liquid obtained in step (3) is centrifuged at 10000 r / min for 20 min, filtered through a 0.22 μm microporous membrane to remove impurities, and the filtrate is collected; the filtrate is pre-frozen at -45℃ for 4 h, and then freeze-dried at a vacuum of 20 Pa and a sublimation temperature of -15℃ until the moisture content does not exceed 5%, and then ground into powder and passed through a 100-mesh sieve to obtain approximately 94 g of freeze-dried powder.

[0042] Example 2:

[0043] Formula ratio: 200g roselle, 80g dandelion, 120g kudzu, 20g Panax notoginseng, 70g Dendrobium officinale, and Lactobacillus plantarum inoculated at 5% of the herbal tea liquid volume, with a bacterial concentration of 1.0 × 10⁻⁶. 9 cfu / mL.

[0044] The preparation method of the embodiment is as follows:

[0045] Step (1) Preparation of flower tea extract: Mix the raw materials of flower tea evenly according to the ratio, add 60% ethanol at a solid-liquid ratio of 1:40 (g / mL), and extract under biological conditions of 70℃, ultrasonic frequency of 40kHZ and power of 100W for 50min to obtain the extract; concentrate the extract under reduced pressure until there is no alcohol taste to obtain flower tea medicinal liquid.

[0046] Step (2) Preparation of probiotic culture: Lactobacillus plantarum was inoculated onto MRS solid medium and cultured under anaerobic conditions at 37℃ for screening and purification; single colonies were picked and inoculated into MRS liquid medium and cultured under anaerobic conditions at 37℃ to obtain Lactobacillus plantarum culture; the effective viable count of the culture was adjusted to 1.0 × 10⁻⁶. 9 cfu / mL;

[0047] Step (3) Preparation of probiotic fermented flower tea liquid: The probiotic liquid obtained in step (2) is inoculated into the flower tea liquid at 5% of the volume of the flower tea liquid obtained in step (1), and fermented under anaerobic conditions at 37℃ for 72 hours to obtain probiotic fermented flower tea liquid.

[0048] Step (4) Preparation of freeze-dried powder: The probiotic fermented flower tea liquid obtained in step (3) is centrifuged at 10000 r / min for 20 min, filtered through a 0.22 μm microporous membrane to remove impurities, and the filtrate is collected; the filtrate is pre-frozen at -45℃ for 4 h, and then freeze-dried at a vacuum of 20 Pa and a sublimation temperature of -15℃ until the moisture content does not exceed 5%, and then ground into powder and passed through a 100-mesh sieve to obtain about 98 g of freeze-dried powder.

[0049] Example 3:

[0050] Formula ratio: 100g roselle, 120g dandelion, 80g kudzu, 40g Panax notoginseng, 70g Dendrobium officinale, and Lactobacillus plantarum inoculated at 5% of the herbal tea liquid volume, with a bacterial concentration of 1.0 × 10⁻⁶. 9 cfu / mL.

[0051] The preparation method of the embodiment is as follows:

[0052] Step (1) Preparation of flower tea extract: Mix the raw materials of flower tea evenly according to the ratio, add 60% ethanol at a solid-liquid ratio of 1:40 (g / mL), and extract under biological conditions of 70℃, ultrasonic frequency of 40kHZ and power of 100W for 50min to obtain the extract; concentrate the extract under reduced pressure until there is no alcohol taste to obtain flower tea medicinal liquid.

[0053] Step (2) Preparation of probiotic culture: Lactobacillus plantarum was inoculated onto MRS solid medium and cultured under anaerobic conditions at 37℃ for screening and purification; single colonies were picked and inoculated into MRS liquid medium and cultured under anaerobic conditions at 37℃ to obtain Lactobacillus plantarum culture; the effective viable count of the culture was adjusted to 1.0 × 10⁻⁶. 9 cfu / mL

[0054] Step (3) Preparation of probiotic fermented flower tea liquid: The probiotic liquid obtained in step (2) is inoculated into the flower tea liquid at 5% of the volume of the flower tea liquid obtained in step (1), and fermented under anaerobic conditions at 37℃ for 72 hours to obtain probiotic fermented flower tea liquid.

[0055] Step (4) Preparation of freeze-dried powder: The probiotic fermented flower tea liquid obtained in step (3) is centrifuged at 10000 r / min for 20 min, filtered through a 0.22 μm microporous membrane to remove impurities, and the filtrate is collected; the filtrate is pre-frozen at -45℃ for 4 h, and then freeze-dried at a vacuum of 20 Pa and a sublimation temperature of -15℃ until the moisture content does not exceed 5%, then ground into powder and passed through a 100-mesh sieve to obtain approximately 82 g of freeze-dried powder.

[0056] Comparative Example 1:

[0057] Formula ratio: 150g roselle, 100g dandelion, 100g kudzu, 30g Panax notoginseng, 90g Dendrobium officinale.

[0058] The preparation method of the embodiment is as follows:

[0059] Step (1) Preparation of flower tea extract: Mix the raw materials of flower tea evenly according to the ratio, add 60% ethanol at a solid-liquid ratio of 1:40 (g / mL), and extract under biological conditions of 70℃, ultrasonic frequency of 40kHZ and power of 100W for 50min to obtain the extract; concentrate the extract under reduced pressure until there is no alcohol taste to obtain flower tea medicinal liquid.

[0060] Step (2) Preparation of freeze-dried powder: The herbal tea liquid obtained in step (1) is centrifuged at 10000 r / min for 20 min, filtered through a 0.22 μm microporous membrane to remove impurities, and the filtrate is collected; the filtrate is pre-frozen at -45℃ for 4 h, and then freeze-dried at a vacuum of 20 Pa and a sublimation temperature of -15℃ until the moisture content does not exceed 5%, and then ground into powder and passed through a 100-mesh sieve to obtain approximately 94 g of freeze-dried powder.

[0061] Example 4: Study on the hepatoprotective and hangover-relieving effects of the herbal tea of ​​the present invention on mice with acute alcohol intoxication.

[0062] Experimental animals: 70 male C57BL / 6 mice, weighing 26-30g, provided by the Animal Center of Nanjing University of Chinese Medicine.

[0063] Dosage setting basis: The recommended daily human dosage for this product is one sachet per day, each sachet containing 5g of lyophilized powder. Based on an average adult weight of 60kg, the daily human dosage is 0.083g of lyophilized powder / kg body weight. According to the human-mouse body surface area conversion factor of 9.1 in *Pharmacological Experimental Methodology*, the equivalent dose for mice is calculated to be 0.76g of lyophilized powder / kg body weight. All example groups and comparative groups used the equivalent mouse dose of 0.76g of lyophilized powder / kg body weight as the dosage. The yield of lyophilized powder in all examples and comparative groups was approximately 20%, therefore the equivalent dose is equivalent to 3.8g / kg of raw drug.

[0064] Test drug: The lyophilized powder prepared by the methods of Examples 1, 2, 3 and Comparative Example 1 of this invention was reconstituted with physiological saline to the required concentration before use. The dosage for each example group and the comparative example group was 0.76 g of lyophilized powder / kg body weight, which was the equivalent dose for mice.

[0065] Positive control drug: Silymarin capsules (National Drug Approval Number H20040299), prepared into a solution of the required concentration with physiological saline before use, with a dosage of 100 mg / kg.

[0066] Drug preparation: Take appropriate amounts of the lyophilized powder prepared by the methods of Examples 1, 2, 3 and Comparative Example 1 of this invention, dissolve and dilute with physiological saline to a concentration of 76 mg lyophilized powder / ml, and prepare immediately before use. Take an appropriate amount of the contents of silymarin capsules, dissolve and dilute with physiological saline to a concentration of 10 mg / mL (positive control group), and prepare immediately before use.

[0067] Animal grouping and administration: After one week of acclimatization, mice were randomly divided into seven groups: a blank control group, a model control group, a positive drug group, a comparative example group 1, an example group 1, an example group 2, and an example group 3, with 10 mice in each group. Administered via gavage once daily for 30 consecutive days, with a volume of 0.1 mL / kg body weight. The blank control group and the model control group received an equal volume of physiological saline; the positive drug group received silymarin solution 100 mg / kg; and the comparative example group, example group 1, example group 2, and example group 3 received 0.76 g / kg of the lyophilized powder prepared in the corresponding example. After the last gavage, except for the blank control group, all other groups were fasted for 16 hours but allowed free access to water, and then administered a single gavage of 12 mL / kg body weight of 50% (v / v) ethanol aqueous solution. Samples were collected 6 hours later. The blank control group was fasted but not administered ethanol via gavage, and samples were collected directly.

[0068] Biological sample collection: Six hours after gavage with 50% ethanol, blood was collected from the eyes of mice in each group and placed in centrifuge tubes. The mice were then euthanized by cervical dislocation. Blood samples were centrifuged at 12000 rpm for 15 minutes at 4°C to separate serum, which was then aliquoted and stored at -20°C for later analysis. Liver tissue was immediately dissected, washed with pre-cooled physiological saline to remove surface blood, blotted dry with filter paper, weighed, and a portion of the liver tissue was fixed in 4% paraformaldehyde fixative for histopathological examination; the remaining liver tissue was cryopreserved at -80°C for biochemical assays.

[0069] Evaluation of Alcohol-Relieving Efficacy - Behavioral Experiment of Intoxication: After oral administration of 50% ethanol to mice in each group, timing was immediately started and the righting reflex test was performed. Judgment Criteria: The mice were turned over and placed in a supine position, and it was observed whether they could spontaneously return to a prone position within 30 seconds. If the mouse remained in a supine position for more than 30 seconds without being able to turn over spontaneously, it was considered to have entered an intoxicated state (loss of righting reflex); if the mouse could spontaneously turn over and return to a prone position within 30 seconds, it was considered to have sobered up (recovery of righting reflex). Recorded indicators were calculated according to the following formula:

[0070] Intoxication time (min) = Time when righting reflex disappears - Time of last gavage

[0071] Sobering time (min) = Righting reflex recovery time - Righting reflex disappearance time

[0072] Success rate of sobering up (%) = (Number of animals successfully sobered up during the observation period / Total number of animals in the group) × 100%

[0073] Evaluation of liver-protective efficacy – Biochemical index testing

[0074] Strictly follow the instructions for each kit and test the following indicators: liver injury markers—serum ALT (alanine aminotransferase) and AST (aspartate aminotransferase) activities; antioxidant indicators—liver SOD (superoxide dismutase) activity, liver MDA (malondialdehyde) content, and liver GSH (reduced glutathione) content.

[0075] Statistical analysis: GraphPad Prism 9.5 software was used for statistical analysis of the experimental data. Quantitative data are expressed as mean ± standard deviation. The results are indicated by the following: Independent samples t-test was used for comparisons between two groups, and Welch-corrected t-test was used when variances were unequal. Fisher's exact test was used to compare the success rate of hangover detoxification. A p-value < 0.05 was considered statistically significant, and a p-value < 0.01 was considered statistically significant.

[0076] The experimental results are shown in Table 1-3 or Figure 1-8 .

[0077] Table 1. Effects of liver-protecting and hangover-relieving herbal teas on the latency period and sobering time of acutely intoxicated mice.

[0078] Model control group 8.0±2.4 171.6±116.4 50 Positive drug group 7.6±2.2 153.9±65.4 70 Comparative Example 1 <![CDATA[10.7±1.9 ** ]]> 128.5±48.0 80 Example 1 <![CDATA[21.7±3.5 **\ΔΔ ]]> <![CDATA[68.5±18.0 **\ΔΔ ]]> 100 Example 2 <![CDATA[15.5±2.5 **\Δ ]]> <![CDATA[95.0±35.0 * ]]> 80 Example 3 <![CDATA[12.3±2.2 ** ]]> 125.0±42.0 70

[0079] Note: Compared with the model control group, *P<0.05, **P<0.01; compared with Comparative Example 1, Δ P < 0.05 ΔΔ P < 0.01. Sobering time was only included in the statistics for individuals who successfully sobered up during the observation period.

[0080] As shown in Table 1, the latency period for alcohol intoxication in the model control group mice was 8.0 ± 2.4 min, and the success rate of sobering up was 50.0%. The positive control drug silymarin did not significantly improve the latency period for alcohol intoxication (7.6 ± 2.2 min, P > 0.05), and the success rate of sobering up was 70.0%, consistent with the pharmacological characteristics of silymarin, which primarily acts on liver protection. The probiotic fermented flower tea freeze-dried powder prepared in Example 1 of this invention significantly prolonged the latency period for alcohol intoxication to 21.7 ± 3.5 min (approximately 171% longer than the model group, P < 0.01), shortened the sobering up time to 68.5 ± 18.0 min (approximately 60% shorter than the model group), and increased the success rate of sobering up to 100.0%. Regarding the sobering up time, due to the small number of successfully sobered individuals in the model group (n = 5) and the large individual differences (SD = 116.4), no statistical significance was achieved between the treatment groups and the model group, but the above data all showed a clear trend towards alcohol relief. Compared with the unfermented Comparative Example 1, the intoxication latency period in Example 1 was prolonged by approximately 103% (P < 0.01), indicating that the fermentation process significantly enhances the hangover relief effect. Among the three example groups, Example 1 showed the best hangover relief effect.

[0081] Table 2. Effects of liver-protecting and hangover-relieving herbal teas on serum ALT and AST levels in acutely intoxicated mice.

[0082] Blank control group 23.0±3.1 162.1±35.2 Model control group <![CDATA[135.6±15.2 ## ]]> <![CDATA[268.9±31.8 ## ]]> Positive drug group <![CDATA[86.9±18.5 ** ]]> <![CDATA[207.3±30.1 ** ]]> Comparative Example 1 <![CDATA[113.8±10.5 ** ]]> <![CDATA[238.5±18.5 ** ]]> Example 1 <![CDATA[51.3±5.5 **\ΔΔ ]]> <![CDATA[152.7±14.5 **\ΔΔ ]]> Example 2 <![CDATA[67.4±8.5 **\ΔΔ ]]> <![CDATA[180.9±12.8 **\ΔΔ ]]> Example 3 <![CDATA[75.3±6.8 **\ΔΔ ]]> <![CDATA[198.5±12.9 **\ΔΔ ]]>

[0083] Note: Compared with the blank control group, # P < 0.05 ## P < 0.01; compared with the model control group, *P < 0.05, **P < 0.01; compared with Comparative Example 1, Δ P < 0.05 ΔΔ P < 0.01. The same applies to the following table.

[0084] As shown in Table 2, the product of this invention can significantly reduce serum ALT and AST levels. In Example 1 group, ALT decreased to 51.3±5.5 U / L (approximately 62% lower than the model group, P<0.01), and AST decreased to 152.7±14.5 U / L (approximately 47% lower than the model group, P<0.01); in Example 2 group, ALT decreased to 67.4±8.5 U / L (P<0.01), and AST decreased to 180.9±12.8 U / L (P<0.01); in Example 3 group, ALT decreased to 75.3±6.8 U / L (P<0.01), and AST decreased to 198.5±12.9 U / L (P<0.01). Compared with the unfermented Comparative Example 1, Example 1 showed a reduction of approximately 55% in ALT (P<0.01) and approximately 36% in AST (P<0.01), indicating that the fermentation process significantly enhanced the hepatoprotective effect.

[0085] Table 3. Effects of liver-protecting and hangover-relieving herbal teas on liver SOD, MDA, and GSH levels in acutely intoxicated mice.

[0086] Blank control group 82.2±32.9 8.3±0.8 740.3±91.2 Model control group <![CDATA[29.9±10.2 ## ]]> <![CDATA[19.6±3.6 # #]]> <![CDATA[545.6±99.8 ## ]]> Positive drug group <![CDATA[67.9±26.8 ** ]]> <![CDATA[7.7±1.3 ** ]]> <![CDATA[815.4±71.9 ** ]]> Comparative Example 1 <![CDATA[51.2±11.2 ** ]]> <![CDATA[12.8±1.4 ** ]]> <![CDATA[682.9+86.5 ** ]]> Example 1 <![CDATA[87.8±9.8 **\ΔΔ ]]> <![CDATA[6.0±1.1 **\ΔΔ ]]> <![CDATA[956.6±98.9 **\ΔΔ ]]> Example 2 <![CDATA[77.5±10.8 **\Δ ]]> <![CDATA[8.0±1.0 **\Δ ]]> <![CDATA[882.7±92.6 **\Δ ]]> Example 3 <![CDATA[71.6±11.2 **\Δ ]]> <![CDATA[9.2±0.9 **\Δ ]]> <![CDATA[853.1±95.8 **\Δ ]]>

[0087] As shown in Table 3, the product of this invention can significantly restore liver SOD activity, reduce MDA content, and increase GSH content. In Example 1 group, SOD recovered to 87.8±9.8 U / mg prot (an increase of approximately 194% compared to the model group, P<0.01), MDA decreased to 6.0±1.1 nmol / mg (a decrease of approximately 69% compared to the model group, P<0.01), and GSH increased to 956.6±98.8 μg / g liver tissue (an increase of approximately 75% compared to the model group, P<0.01). All indicators recovered to levels close to the blank control group, comparable to or slightly better than the positive control drug silymarin. Compared with the unfermented Comparative Example 1, Example 1 showed an increase of approximately 71% in SOD (P<0.01), a decrease of approximately 53% in MDA (P<0.01), and an increase of approximately 40% in GSH (P<0.01), further confirming that the fermentation process can effectively enhance the antioxidant capacity of the herbal tea composition.

[0088] Depend on Figure 8 The results of pathological observation of liver tissue showed that the model group had typical pathological changes such as inflammatory infiltration, fatty degeneration, and cell necrosis in the liver tissue. The sample of Example 1 could significantly improve the abnormal pathological morphology of liver tissue induced by ethanol and reduce the degree of fatty degeneration, inflammatory cell infiltration and necrosis of hepatocytes.

[0089] Experimental Conclusion: Based on the above experimental results, the probiotic fermented liver-protecting and hangover-relieving freeze-dried herbal tea powder of this invention showed significant liver-protecting and hangover-relieving effects in a mouse model of acute alcoholic liver injury.

[0090] Regarding liver protection, the product of this invention can significantly reduce serum ALT and AST levels, restore liver SOD activity, reduce MDA content, and increase GSH levels. It exerts its liver-protective effect through multiple pathways, including protecting hepatocyte membrane integrity and restoring the antioxidant defense system. In Example 1, all indicators returned to near-normal levels, showing no significant difference from the positive control group.

[0091] Regarding hangover relief, the product of this invention can significantly prolong the incubation period of intoxication, shorten the sobering time, and improve the success rate of sobering up, with the first group of examples showing the best hangover relief effect.

[0092] In terms of process innovation, probiotic fermentation is a key technological innovation for enhancing product efficacy. Under the same formula, all indicators of the fermented group were significantly better than those of the unfermented group, confirming that fermentation can effectively enhance the hangover-relieving and liver-protecting activities of the herbal tea formula. Regarding the formulation ratio, Example 1 has the most appropriate and optimal formula due to the synergistic effect of clearing heat and cooling blood, relieving hangover and invigorating the spleen, and nourishing yin and protecting the liver.

Claims

1. A probiotic fermented flower tea composition for liver protection and hangover relief, characterized in that, The composition is obtained by fermenting a flower tea extract with probiotics. The flower tea extract consists of the following flower teas in parts by weight: 10-20 parts roselle, 8-12 parts dandelion, 8-12 parts kudzu, 2-4 parts Panax notoginseng, and 7-11 parts Dendrobium officinale. The probiotic is Lactobacillus plantarum.

2. The probiotic fermented flower tea composition for liver protection and hangover relief according to claim 1, characterized in that, The composition is obtained by fermenting a flower tea extract with probiotics. The flower tea extract consists of the following flower teas in parts by weight: 15 parts roselle, 10 parts dandelion, 10 parts kudzu, 3 parts Panax notoginseng, and 9 parts Dendrobium officinale. The probiotic is Lactobacillus plantarum, accession number CGMCC15013.

3. The probiotic fermented flower tea composition for liver protection and hangover relief according to claim 1, characterized in that, The effective live bacteria count of the probiotics is 1.0 × 10⁻⁶. 7 cfu / mL~1.0×10 9 cfu / mL, preferably 1.0 × 10⁻⁶ 9 cfu / mL.

4. A freeze-dried powder, characterized in that, The freeze-dried powder is prepared by freeze-drying the probiotic fermented flower tea composition according to any one of claims 1 to 3.

5. The freeze-dried powder according to claim 4, characterized in that, The preparation method includes the following steps: Step (1) Preparation of flower tea extract: Mix the raw materials of flower tea evenly according to the ratio, add 60% ethanol at a solid-liquid ratio of 1:40 (g / mL), and extract under biological conditions of 70℃, ultrasonic frequency of 40kHZ and power of 100W for 50min to obtain the extract; concentrate the extract under reduced pressure until there is no alcohol taste to obtain flower tea medicinal liquid. Step (2) Preparation of probiotic culture: Lactobacillus plantarum was inoculated onto MRS solid medium and cultured under anaerobic conditions at 37℃ for screening and purification; single colonies were picked and inoculated into MRS liquid medium and cultured under anaerobic conditions at 37℃ to obtain Lactobacillus plantarum culture; the effective viable count of the culture was adjusted to 1.0 × 10⁻⁶. 9 cfu / mL; Step (3) Preparation of probiotic fermented flower tea liquid: The probiotic liquid obtained in step (2) is inoculated into the flower tea liquid at 5% of the volume of the flower tea liquid obtained in step (1), and fermented under anaerobic conditions at 37℃ for 72 hours to obtain probiotic fermented flower tea liquid. Step (4) Preparation of freeze-dried powder: The probiotic fermented flower tea liquid obtained in step (3) is centrifuged at 10000 r / min for 20 min, filtered through a 0.22 μm microporous membrane to remove impurities, and the filtrate is collected; the filtrate is pre-frozen at -45℃ for 4 h, and then freeze-dried at a vacuum of 20 Pa and a sublimation temperature of -15℃ until the moisture content does not exceed 5%, then ground into powder and passed through a 100-mesh sieve to obtain freeze-dried powder.

6. The use of the probiotic fermented herbal tea composition according to any one of claims 1 to 3 or the freeze-dried powder according to claim 4 in the preparation of products with liver-protecting and / or hangover-relieving effects.

7. The application according to claim 6, characterized in that, The product in question is a pharmaceutical, health product, or food.

8. The application according to claim 7, characterized in that, The dosage of the probiotic fermented flower tea composition or freeze-dried powder in the mouse model of the product is 0.76 g / kg based on the freeze-dried powder, which is equivalent to 3.8 g / kg of raw drug.

9. The application according to claim 6, characterized in that, The liver-protective effects include reducing serum alanine aminotransferase (ALT) and / or aspartate aminotransferase (AST) levels in individuals with alcoholic liver injury, and / or increasing liver superoxide dismutase (SOD) activity and / or glutathione (GSH) content, and / or reducing liver malondialdehyde (MDA) content. The hangover-relieving effects include prolonging the latency period of intoxication and / or increasing the success rate of sobering up and / or shortening the sobering up time.

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