Five-flavor mannan compound preparation based on probiotic fermentation and preparation method of five-flavor mannan compound preparation

Abstract

The invention provides a preparation method of a five-flavor sweet dew fermented liquid medicine based on probiotics lactobacillus casei S8, and relates to the field of traditional Chinese medicine fermentation and external preparations. According to the method, the compatibility ratio (1: 1: 2: 2: 3) of five Tibetan medicinal materials including juniperus formosana leaves, rhododendron parvifolium, myricaria laxiflora, Tibetan ephedra and artemisia sieversiana is optimized, lactobacillus casei S8 is inoculated on the basis of decoction extraction for temperature-controlled and oxygen-controlled fermentation, the release efficiency of functional components such as total flavonoids, polyphenols and lactic acid is remarkably improved, the pH value of fermentation liquor is stabilized at 4.0-4.5, and the fermentation liquor is suitable for industrial production. Good skin adaptability and oxidation resistance are realized. Compared with traditional distiller's yeast fermentation, the process is free of alcohol by-products, stable in strain, controllable in process and suitable for development of various dosage forms such as bath, spray, effervescent tablets and wet compress liquid. The method has novelty, practicability and good industrialization prospects.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine fermentation and its applications, and particularly to a preparation method of a compound preparation of five-flavor manna with probiotics as fermentation strains, and in particular to a probiotic-fermented five-flavor manna preparation and its preparation process for improving the bioavailability of traditional Chinese medicine and expanding the application form. Background Art

[0002] Five-flavor manna is a classic prescription in traditional Tibetan medicine medicated bath, originating from ancient medical books such as "Four Medical Tantras", "Thousands of Sarira", and "Blue Beryl", and is widely used in the external treatment of diseases such as regulating qi and blood, dispelling wind and dampness, and relaxing tendons and activating collaterals. It is mainly composed of five Tibetan medicinal materials, namely Juniperus communis, Rhododendron anthopogonoides, Myricaria bracteata, Ephedra saxatilis, and Artemisia sieversiana, and is formulated in a mass ratio of 1:1:2:2:3, with multiple pharmacological effects such as anti-rheumatic, meridians-relaxing, heat-clearing and detoxifying, and fatigue-relieving.

[0003] The traditional way of using five-flavor manna is mainly medicated bath. In recent years, various fermentation preparation processes have been developed to improve the release and permeability of drug efficacy. Among them, the traditional method represented by Tibetan wine koji fermentation can improve the dissolution of active ingredients such as flavonoids and polyphenols in the medicinal liquid, but there are the following problems: First, the source of the microbial community is complex, the strains are uncontrollable, and the stability of the medicinal liquid components between batches is poor; second, by-products such as alcohol are generated during the fermentation process, which may cause irritation or usage restrictions to the skin or specific populations (such as children and pregnant women); third, there is a lack of systematic research on the correlation between modern microecology and drug efficacy, making it difficult to explain its mechanism of action.

[0004] Probiotic fermentation, as a modern improvement path for traditional Chinese medicine preparations, shows great potential in improving the utilization rate of active ingredients of traditional Chinese medicine, improving flavor and irritation due to its high safety, clear metabolites, and strong process controllability. During the fermentation process, probiotics can secrete substances such as organic acids, polysaccharides, B vitamins, and phenolic acids, which can undergo synergistic transformation reactions with traditional Chinese medicine components, helping to improve the solubility and bioavailability of drug efficacy substances.

[0005] Lactobacillus casei S8 is a dominant probiotic strain with a clear source and stable performance, having strong acid and bile salt tolerance, and being able to stably survive and reproduce in various herbal fermentation systems. During the fermentation process, this strain can efficiently produce lactic acid and low-molecular-weight polysaccharides, which help to regulate the pH of the medicinal liquid, improve skin friendliness, and can promote the release and structural transformation of flavonoids and phenolic compounds, showing good antioxidant, antibacterial, and anti-inflammatory potential, and has been widely used in the fields of food and traditional Chinese medicine fermentation.

[0006] Therefore, constructing a standardized, alcohol-free, and highly safe preparation process for fermenting Wumei Ganlu based on Lactobacillus casei S8 can not only make up for the defects of koji fermentation but also improve product quality and expand the scope of application, which is an important path to promote the modernization and large-scale production of Tibetan medicine. Summary of the Invention

[0007] The purpose of the present invention is to propose a fermentation process for processing and preparing Wumei Ganlu liquid medicine based on the probiotic Lactobacillus casei S8. On the basis of inheriting the classic compatibility and efficacy of the traditional Tibetan medicine bath formula "Wumei Ganlu", a modern probiotic fermentation technology process with strong controllability, high safety, and significantly improved biological potency is established to solve technical problems such as complex flora, uncertain alcohol by-products, and poor quality stability during the traditional koji fermentation process, and to meet diverse application requirements such as traditional Chinese medicine bath therapy, health care, and skin conditioning.

[0008] The present invention selects Juniperus communis, Rhododendron anthopogonoides, Myricaria bracteata, Ephedra saxatilis, and Artemisia sieversiana as the main medicinal materials, and formulates them in a ratio of 1:1:2:2:3. Combining the excellent fermentation and transformation ability and lactic acid metabolism advantage of Lactobacillus casei S8, a new Wumei Ganlu fermentation system that meets the requirements of modern production and pharmacodynamic research is constructed. On the basis of ensuring the traditional efficacy, significant release of pharmacodynamic substances, pH-friendly regulation, and diversified transformation of preparations are achieved.

[0009] To achieve the above purpose, the preparation process of a probiotic Lactobacillus casei S8 fermenting Wumei Ganlu liquid medicine of the present invention includes the following steps:

[0010] (1) Weighing of medicinal materials: Weigh Juniperus communis, Rhododendron anthopogonoides, Myricaria bracteata, Ephedra saxatilis, and Artemisia sieversiana in sequence according to the mass ratio of 1:1:2:2:3 for later use.

[0011] (2) Soaking pretreatment: Place the above-mentioned medicinal materials in purified water and soak them for 1 - 2 hours according to the mass ratio of water to medicinal materials of 3 - 5:1 to fully swell the medicinal materials, which is beneficial for subsequent component extraction.

[0012] (3) Preliminary decoction extraction: Heat and decoct the soaked medicinal materials, first bring to a boil over high heat and then decoct over low heat for 30 - 60 minutes to extract the main water-soluble components such as flavonoids and polyphenols; stop heating after the water level drops to the same level as the medicine surface, and cool to 30 - 37 °C for standby.

[0013] (4) Inoculation and fermentation: Add the active probiotic Lactobacillus casei S8 to the medicinal liquid cooled to the target temperature. The inoculation ratio is 2–5% of the mass of the fermentation broth, and the initial bacterial liquid concentration is 1×10 7 –10 8 CFU / mL. At the same time, adjust the pH to 5.5–6.0.

[0014] (5) The fermentation control conditions are as follows: · Control the fermentation temperature at 35–37°C; · Maintain the dissolved oxygen DO in the tank at 10–25%; · The optional ventilation ratio is 0.1–0.5 vvm; · Control the stirring speed at 60–120 r / min (or static fermentation); · Control the fermentation cycle within 48–72 hours.

[0015] (6) The criteria for judging the fermentation end point are as follows: The pH of the fermentation broth drops to 4.0–4.5, the lactic acid concentration reaches 1–2.5 g / L, the increase in the total flavonoid content is ≥20%, and the sensory color is uniform, without peculiar smell and precipitation.

[0016] (7) Post-fermentation treatment: · Centrifuge at 2000–8000 r / min for 5–20 min to remove the precipitate; · Conduct microfiltration with a filter membrane pore size of 0.3–0.5 μm; · Control the sterilization pressure at 250–320 Mpa (if using high-pressure sterilization); · Obtain the supernatant as the final fermented liquid product of Wuweiganlu.

[0017] The Lactobacillus casei S8 can enhance its fermentation efficiency through pre-adaptation culture. The specific method is as follows: Mix the primary decoction of Wuweiganlu and highland barley powder in a mass ratio of 5∶1 to form a composite medium, inoculate the bacterial liquid and culture until the OD value ≥2, and then use it for fermentation inoculation, which helps to improve the survival ability and efficacy transformation efficiency of the strain in the herbal fermentation matrix.

[0018] The final medicinal liquid can be prepared into the following different dosage forms according to needs: · Liquid preparation: Used for bath, foot bath, and medicinal liquid spray; · Solid preparation: Such as effervescent tablets, freeze-dried powders, and tablet granules; · External products: Such as wet compress liquids, cold compress gels, and beauty mask carriers, etc.

[0019] The probiotic-fermented Wuweiganlu medicinal liquid of the present invention can significantly improve the following efficacy-related indicators: · The total flavonoid extraction rate can be increased to more than 40–50%; · Generate new metabolites such as lactic acid and soluble polysaccharides; · Lower the pH to a skin-friendly range (4.0–4.5); · Through preliminary biochemical tests, the antioxidant capacity is increased by about 1.5–2 times.

[0020] Compared with the traditional koji fermentation method, the present invention has the following remarkable advantages: 1. Using standardized probiotic strains (Lactobacillus casei S8), with strong controllability and stable fermentation process; 2. No alcohol by-products are generated, safer to use, suitable for children, pregnant women, and people with sensitive skin; 3. Can improve the content of key components of Wuweiganlu such as flavonoids, polyphenols, and lactic acid, and improve the penetration and absorption efficiency; 4. The pH is naturally regulated during the fermentation process, reducing the dependence on externally added acid and base agents, and having better skin adaptability; 5. Can achieve large-scale, continuous, and modular production, with good batch-to-batch consistency and excellent cost control; 6. The product has various forms, meeting the growing market demand for traditional Chinese medicine external use / tonic products.

[0021] Therefore, the present invention has established a Wuweiganlu probiotic fermentation system that conforms to the development trend of modern traditional Chinese medicine preparations, integrating the advantages of classic Tibetan medicine bath prescriptions and modern microecological fermentation, providing a new solution for the modern production, efficacy improvement, and multi-formulation development of traditional Chinese medicine compounds, and having broad clinical transformation value and market application prospects.

[0022] The highland barley flour has multiple functional roles in the fermentation system: one is as a fermentable carbon source for the probiotic Lactobacillus casei S8, which can promote its rapid reproduction and lactic acid production; the other is that it contains rich dietary fiber and soluble polysaccharides, which helps to improve the consistency and stability of the fermentation broth and form a culture medium suitable for the growth of microecology; thirdly, the highland barley itself contains a certain amount of phenolic acids, polyphenols, and B vitamins, which act together with traditional Chinese medicine components and may synergistically enhance the antioxidant and immunomodulatory activities of the preparation. Therefore, the highland barley flour is not only a substrate for providing nutrition, but also plays an important role in regulating the texture, enhancing the efficacy, and improving the flavor in the fermentation system. Detailed implementation methods

[0023] Example 1

[0024] The preparation and fermentation process of the Wuweiganlu medicinal liquid fermented by Lactobacillus casei S8 according to the present invention is as follows:

[0025] (1) Weigh the medicinal materials: Weigh Juniperus sabina L., Rhododendron parvifolium Adams, Myricaria bracteata Royle, Ephedra saxatilis Royle ex Florin, and Artemisia sieversiana Ehrhart ex Willd. according to the mass ratio of 1∶1∶2∶2∶3 respectively, and mix them thoroughly for later use.

[0026] (2) Immersion extraction: Put the medicinal materials into purified water, soak for 1 hour according to the mass ratio of water to medicinal materials of 5:1; then heat to boiling, decoct with slow fire for 60 minutes, extract the medicinal liquid, and cool to 35 °C for standby.

[0027] (3) Inoculation and fermentation: Add the active dry Lactobacillus casei S8 bacterial liquid to the medicinal liquid cooled to 35 °C, and the inoculation ratio is 2% of the volume of the medicinal liquid, with an initial bacterial liquid concentration of 1×10 8 CFU / mL, and adjust the initial pH to 5.5.

[0028] (4) Fermentation process: In a sealed constant-temperature fermentation container, carry out static fermentation at 35 °C for 48 hours, during which maintain the dissolved oxygen DO at 15%, and there is no stirring and aeration operation.

[0029] (5) Judgment of fermentation end point: When fermentation ends, the lactic acid concentration is 1.2 g / L, the pH drops to 4.2, and by HPLC determination, the total flavonoid extraction rate is increased to 42.3%.

[0030] (6) Post-treatment: Centrifuge the fermentation broth at 2000 r / min for 5 min, carry out microfiltration treatment with a filter membrane pore size of 0.3 μm, with a filtration pressure of 0.4 Mpa and a sterilization pressure of 250 Mpa, and take the supernatant as the Wuweiganlu probiotic fermentation medicinal liquid.

[0031] Example 2

[0032] On the basis of Example 1, this example adjusted the fermentation temperature, stirring rate and the addition ratio of highland barley powder. The specific process is as follows:

[0033] (1) Weigh the medicinal materials: Weigh Juniperus Sabina L., Rhododendron parvifolium Adams, Myricaria bracteata Royle, Ephedra saxatilis Royle ex Florin, Artemisia sieversiana Ehrhart ex Willd. according to the mass ratio of 1:1:2:2:3 respectively, mix well and set aside.

[0034] (2) Immersion extraction: Put the medicinal materials into purified water, soak for 1.5 hours according to the mass ratio of water to medicinal materials of 4:1; then heat to boiling, decoct with slow fire for 50 minutes, extract the medicinal liquid, and cool to 36 °C for standby.

[0035] (3) Prepare the culture medium: Mix the Wuweiganlu decoction and highland barley powder according to the mass ratio of 5:1 to form a composite culture medium, and inoculate the Lactobacillus casei S8 bacterial liquid for pre-culture for 12 hours.

[0036] (4) Inoculation and fermentation: Inoculate the pre-cultured bacterial liquid at 3% of the volume of the medicinal liquid, with an initial bacterial liquid concentration of 1×10 8 CFU / mL, and adjust the initial pH to 5.8.

[0037] (5) Fermentation process: Carry out low-speed stirring fermentation in a fermenter, control the temperature at 36 °C, the stirring rate at 60 r / min, the aeration rate at 0.2 vvm, the fermentation time at 60 hours, and maintain DO at 20%.

[0038] (6) Determination of fermentation end point: At the end of fermentation, the lactic acid concentration is 1.5 g / L, the pH drops to 4.1, and the total flavonoid content is 45.7%.

[0039] (7) Post-treatment: The fermentation broth is centrifuged at 4000 r / min for 10 min, microfiltered through a 0.3 μm filter membrane, the filtration pressure is 0.5 Mpa, and the sterilization pressure is 280 Mpa to obtain the fermented liquid of Wuweiganlu probiotics.

[0040] Example 3

[0041] In this example, the stirring intensity and fermentation time were optimized, and the inoculation ratio and sterilization pressure were adjusted. The specific process is as follows:

[0042] (1) Weigh Juniperus sabina L., Rhododendron parvifolium Adams, Myricaria bracteata Royle, Ephedra saxatilis Royle ex Florin, Artemisia sieversiana Ehrhart ex Willd. according to the mass ratio of 1:1:2:2:3, mix them evenly and put them into water, and soak for 2 hours according to 4:1.

[0043] (2) Decoction extraction: Decoct with slow fire for 30 minutes and then cool to 37 °C.

[0044] (3) Inoculation: After pre-culturing Lactobacillus casei S8, inoculate it, the inoculation ratio is 5%, the initial bacterial concentration is 1×10 8 CFU / mL, and the initial pH is 5.0.

[0045] (4) Fermentation conditions: The temperature is 37 °C, the stirring speed is 120 r / min, the aeration ratio is 0.3 vvm, ferment for 72 hours, and maintain the DO at 25%.

[0046] (5) Fermentation end point: The lactic acid concentration is 2.1 g / L, the pH drops to 4.0, and the total flavonoid extraction rate is 49.3%.

[0047] (6) Post-treatment: Centrifuge at 6000 r / min for 10 min, filter through a 0.5 μm filter membrane, and the sterilization pressure is 300 Mpa to obtain the fermented product.

[0048] Example 4

[0049] In this example, high-temperature fermentation conditions and medium aeration were used to verify the stability of the strain and the release of flavonoids.

[0050] (1) Weigh the Wuweiganlu medicinal materials, and the ratio of Juniperus sabina L., Rhododendron parvifolium Adams, Myricaria bracteata Royle, Ephedra saxatilis Royle ex Florin, Artemisia sieversiana Ehrhart ex Willd. is 1:1:2:2:3, soak for 1 hour, and the water-to-material ratio is 5:1.

[0051] (2) Decoct: Decoct with slow fire for 60 minutes and cool to 40 °C.

[0052] (3) Inoculation: The access ratio of the live bacteria solution of Lactobacillus casei S8 is 4%, and the initial pH value is 5.5.

[0053] (4) Fermentation process: The fermentation temperature is 40°C, stirring speed is 80 r / min, aeration rate is 0.2 vvm, and the fermentation cycle is 60 hours.

[0054] (5) Endpoint detection: The lactic acid production is 1.7 g / L, pH drops to 4.3, and the total flavonoid content is 47.0%.

[0055] (6) Post-treatment: Centrifuge at 8000 r / min for 15 min, filter through a 0.3 μm filter membrane, and the sterilization pressure is 320 Mpa.

[0056] Example 5

[0057] This example uses the lowest proportion of highland barley flour and neutral fermentation temperature to control the viscosity and fermentation flavor. The process is as follows:

[0058] (1) The proportion of medicinal materials is Juniperus sabina L., Rhododendron parvifolium Adams, Myricaria bracteata Royle, Ephedra saxatilis Royle ex Florin, Artemisia sieversiana Ehrh. = 1:1:2:2:3, soaking time is 2 hours, and the water-to-material ratio is 3:1.

[0059] (2) Decoction extraction: Simmer over low heat for 45 minutes and cool to 33°C.

[0060] (3) The inoculation ratio is 3%, and the initial pH is 5.7.

[0061] (4) Fermentation conditions: Temperature is 33°C, static fermentation for 48 hours, without aeration and stirring, and DO is controlled at 10%.

[0062] (5) Fermentation endpoint: Lactic acid concentration is 1.0 g / L, pH drops to 4.4, and total flavonoid is 41.2%.

[0063] (6) Post-treatment: Centrifuge at 2000 r / min for 10 min, microfiltrate through a 0.3 μm filter, and the sterilization pressure is 260 Mpa to obtain the final medicinal liquid.

[0064] The preparation process of the fermented Wuweiganlu medicinal liquid by Lactobacillus casei S8 proposed by the present invention combines classical Tibetan medicine compatibility and modern probiotic fermentation technology, and has the following remarkable technical advantages and practical values:

[0065] 1. Strong process stability: Using Lactobacillus casei S8 with clear source and stable properties as the fermentation strain, avoiding problems such as complex flora and uncontrollable fermentation results in traditional koji, the preparation process can be implemented in a standardized and large-scale manner, improving the product quality consistency and traceability.

[0066] 2. High ingredient release efficiency: Examples 1-5 all show that for the fermentation broth prepared by the process of the present invention, the total flavonoid extraction rate can reach 41.2-49.3%, which is significantly improved compared with the traditional water decoction method, indicating that this fermentation technology can promote the dissolution and transformation of medicinal ingredients such as flavonoids and polyphenols in Wuweiganlu.

[0067] 3. Superior safety and applicability: No alcohol is generated during the fermentation process, avoiding potential irritation problems for the skin or special populations. It is suitable for children, pregnant women, people with sensitive skin, etc., expanding the scope of use of the medicinal liquid and its commercial application scenarios.

[0068] 4. Good microecological regulation: Lactobacillus casei S8 produces appropriate amounts of lactic acid and low-molecular polysaccharides during fermentation, and the pH of the fermentation broth can naturally drop to a skin-friendly range of 4.0-4.5, enhancing skin adaptability and comfort during use.

[0069] 5. Synergistic effect of auxiliary ingredients: As an auxiliary culture substrate, highland barley powder not only provides a carbon source to promote the reproduction of probiotics and the generation of metabolites, but also contains various soluble dietary fibers, vitamins and trace phenols, which can synergistically play antioxidant and conditioning roles with traditional Chinese medicines.

[0070] 6. Support for multi-form applications: The obtained fermentation broth has good stability and can be further developed into liquid preparations (bathing solutions, sprays), solid preparations (effervescent tablets, freeze-dried powders), external products (wet compresses, mask substrates, etc.) to meet the needs of the modernization of traditional Chinese medicines and cross-scenario transformation.

[0071] 7. Outstanding efficacy performance: Experimental data show that the lactic acid concentration in the fermentation broth prepared in the examples is stable at 1.0-2.1 g / L, the total flavonoid content is significantly increased, and it has good antioxidant potential. The preliminary DPPH scavenging rate test shows that its free radical scavenging ability is about 1.5-2 times higher than that of the traditional decoction, laying a foundation for extended functions such as fatigue relief and anti-inflammatory and soothing effects.

[0072] In summary, the present invention improves the traditional Tibetan medicine formula "Wuweiganlu" through a scientific fermentation path, achieving significant improvements in terms of safety, ingredient release, formulation adaptability and function expansion, and having good industrialization prospects and development value for traditional Chinese medicine bath products. Functional application performance

[0073] Based on the Wuweiganlu probiotic fermentation medicinal liquid prepared by the process of the present invention, combined with the existing experience in herbal medicine bath applications, through preliminary user trials and sensory evaluation feedback, it shows the following application characteristics:

[0074] 1. Good skin adaptability: During the bathing process, there are no obvious skin irritations, itching or burning sensations, and the pH value is stable at 4.1-4.4, showing mild and skin-friendly characteristics, suitable for people with sensitive skin;

[0075] 2. Significant fatigue relief: Most testers reported reduced body soreness and improved mental state after 5-7 consecutive days of bathing, with symptoms such as falling asleep faster and waking up more energetic in the morning.

[0076] 3. The liquid has a uniform color, aroma and shape: after lactic acid fermentation, the liquid appears light yellow to brownish yellow, exudes a natural herbal fermentation aroma, has no irritating odor, is visually pleasing, and has high user acceptance;

[0077] 4. Good experience in using effervescent tablets / sprays: After conversion to effervescent tablets, liquid sprays and other product forms, the product still maintains good solubility and functional stability. Users report that it is easy to use, absorbs quickly, and has a soft odor.

[0078] The above-mentioned product effects show that the fermented medicinal liquid prepared by the present invention not only reflects the advantage of enrichment of effective ingredients in terms of chemical composition, but also has good experience, safety and functional performance in actual use, which is in line with the product development trends in the directions of modern cosmeceuticals, health care and family health management.

[0079] In summary, the Wuwei Ganlu probiotic fermentation preparation described in the present invention uses Lactobacillus casei S8 as the functional strain. By optimizing the ratio of medicinal materials, controlling fermentation parameters and introducing composite culture medium regulation means, it not only overcomes the problems of unstable bacterial flora and uncontrollable alcohol by-products in the traditional Tibetan wine koji fermentation process, but also improves the release rate and conversion efficiency of functional ingredients such as total flavonoids, polyphenols, and lactic acid, and significantly enhances the bioavailability and skin adaptability of the final medicinal solution.

[0080] The preparation results and preliminary application feedback from Examples 1-5 demonstrate that the probiotic-fermented Wuwei Ganlu medicinal liquid obtained in this invention exhibits excellent ingredient stability, adjustability, and industrial suitability. It demonstrates promising functional potential in liquid formulations (baths, sprays), solid formulations (effervescent tablets, lyophilized powders), and topical skin carriers (dressings, gels), demonstrating the feasibility of transitioning from traditional medicinal baths to multi-modal modern product development.

[0081] The probiotic fermentation technology system and supporting preparation process constructed by the present invention are a modern bioprocess redevelopment of the medicinal form of the traditional Tibetan medicine "Five Flavors Ganlu". It is novel, creative and has significant practical value. It meets the basic technical conditions for invention patent applications and can be widely used in many industries such as Chinese medicine fermentation preparations, external care products, health care and conditioning, and has good market promotion prospects and intellectual property protection value.

Claims

1. A preparation method of a five - flavor manna probiotic fermentation liquid, characterized in that, The preparation method comprises the following steps: (1) Medicinal material compatibility: Weigh Juniperus communis, Rhododendron anthopogonoides, Myricaria bracteata, Ephedra saxatilis, and Artemisia sieversiana according to the mass ratio of 1:1:2:2:

3. (2) Soaking and decocting: Add the medicinal materials into purified water and soak for 1 - 2 hours according to the water-to-material ratio of 3 - 5:1, then decoct for 30 - 60 minutes and cool to 30 - 37°C. (3) Probiotic inoculation: Active Lactobacillus casei S8 was inoculated into the cooled liquid medicine, and the inoculation amount was 2–5% of the volume of the liquid medicine, with an initial bacterial concentration of 1×10 7 –10 8 CFU / mL, and the initial pH was adjusted to 5.5–6.0; (4) Controlling fermentation conditions: Ferment at 35 - 37°C for 48 - 72 hours, with an aeration ratio of 0.1 - 0.5 vvm, a stirring rate of 60 - 120 r / min or static, and maintain the DO at 10 - 25%. (5) Fermentation termination criteria: When the pH drops to 4.0 - 4.5, the lactic acid concentration is 1.0 - 2.5 g / L, and the total flavonoid extraction rate increases by ≥20%. (6) Post-treatment steps: Centrifuge the fermentation broth at 2000 - 8000 r / min for 5 - 20 min, filter it through a 0.3 - 0.5 μm filter membrane, control the sterilization pressure at 250 - 320 Mpa, and take the supernatant as the finished product.

2. The preparation method according to claim 1, wherein, Before the step (3), it further includes mixing the first decoction of Wuweiganlu with highland barley flour according to the mass ratio of 5:1 and performing strain pre-adaptation culture for 12 hours to improve the activity and stability of Lactobacillus casei S8 in the medicinal liquid.

3. The preparation method according to claim 1, wherein The obtained fermentation broth can be used to prepare various dosage forms, including: (1) Liquid preparations, such as bath liquids, spray liquids, and foot bath liquids; (2) Solid preparations, such as effervescent tablets, freeze-dried powders, and tablet granules; (3) External products, such as cold compress gels, wet compress liquids, and carrier liquids for beauty masks.

4. The medicinal liquid prepared by the fermentation method according to claim 1 or 2, characterized in that, The total flavonoid extraction rate is 40 - 50%, the lactic acid concentration is 1.0 - 2.1 g / L, and the pH range is 4.0 - 4.5, having good skin adaptability and antioxidant properties.

5. A traditional Chinese medicine external composition prepared from the liquid medicine according to any one of claims 1-4, characterized in that, The composition contains the fermentation broth of Wuweiganlu as the main functional ingredient, supplemented with stabilizers, thickeners, or excipients for preparing external care products or health care preparations.