Kidney and liver nourishing compound probiotic cistanche fermented drink and preparation method thereof

Abstract

This invention discloses a kidney-tonifying and liver-nourishing compound probiotic Cistanche fermented drink and its preparation method, comprising the following steps: mixing and sterilizing Cistanche deserticola, Poria cocos, malt, functional components, and natural selenium-strontium mineral water to obtain a fermentation substrate; activating *Lactobacillus casei*, *Lactobacillus acidophilus*, *Lactobacillus rhamnosus*, and *Lactobacillus plantarum* strains respectively, centrifuging and freeze-drying, and then mixing them to obtain a compound probiotic powder; culturing the compound probiotic powder in MRS medium to obtain a compound probiotic liquid; inoculating the compound probiotic liquid into the fermentation substrate for fermentation, sterilizing, and obtaining the kidney-tonifying and liver-nourishing compound probiotic Cistanche deserticola fermented drink. This invention, through the innovative integrated formulation technology of combining Cistanche deserticola, malt, and Poria cocos as medicinal and edible ingredients, multifunctional factor compounding, and compound probiotic fermentation, prepares a kidney-tonifying probiotic Cistanche deserticola fermented drink with high active ingredients, clear efficacy, and excellent flavor.

Description

Technical Field

[0001] This invention relates to the field of microbial fermentation technology, specifically to a compound probiotic Cistanche deserticola fermented drink for tonifying the kidneys and nourishing the liver, and its preparation method. Background Technology

[0002] Developing products with the function of "tonifying the liver and kidneys and replenishing essence and blood" using Cistanche deserticola as the core ingredient is currently a research hotspot targeting men's health and beauty needs. However, existing technologies still face a series of key bottlenecks in the development of such products, restricting their efficacy and market competitiveness:

[0003] (1) Low dissolution rate and low bioavailability of active ingredients: The core active ingredients of Cistanche deserticola (such as echinacoside and verbascoside) and Poria cocos β-(1-3)-glucan are both encapsulated in the dense plant cell wall. Traditional extraction processes have limited cell wall breaking efficiency, resulting in insufficient dissolution of target ingredients and low extraction rate. In addition, the molecular weight of the extract is large, and the direct absorption and utilization rate (bioavailability) by the human body is not high, resulting in waste of raw materials and insufficient actual efficacy.

[0004] (2) The contradiction between unsuitable raw material flavor and artificial flavoring is prominent: The special smell of Cistanche deserticola directly affects the palatability of the product. Adding a lot of sugar or flavoring to cover up the odor is not in line with the trend of clean labeling and deviates from the positioning of healthy products.

[0005] (3) Poor matching of existing fermentation technology “strain-substrate”: Currently, the probiotic fermentation technology applied to food and medicine homology raw materials mostly uses general strains or simple compounding, lacking the ability to target specific components (such as phenylethanol glycosides and polysaccharides) in the Cistanche deserticola system, resulting in limited synergistic effect of the fermentation process on the target active ingredients, and failing to achieve the design goal of “strain-substrate synergy and enhanced efficacy”.

[0006] (4) Lack of systematic optimization of process and unstable product quality: Key parameters such as fermentation temperature, time and inoculation amount rely on experience to set, and are not systematically and data-driven optimized according to the characteristics of compound raw materials and the metabolic law of microbial community. This can easily cause problems such as fluctuations in the content of active ingredients and unstable number of live bacteria between batches, making it difficult to ensure the quality uniformity and functional reliability of large-scale production.

[0007] (5) The formula lacks functional synergistic components and has insufficient comprehensive value. Traditional formulas only contain medicinal and edible raw materials without the addition of functional components, which cannot enhance the effects of tonifying the kidney and liver, anti-fatigue and anti-oxidation. Furthermore, there is no systematic optimization, and there is no synergistic mechanism between active ingredients, probiotics and functional components, resulting in poor product stability and efficacy consistency. Summary of the Invention

[0008] This invention addresses the shortcomings of existing technologies by providing a compound probiotic Cistanche fermented beverage for tonifying the kidneys and liver, and its preparation method. It optimizes specific ratios of fermentation substrate, compound probiotic fermentation system, and fermentation process to prepare a compound probiotic Cistanche fermented beverage for tonifying the kidneys and liver with good flavor and taste, high live bacteria count, stable functional quality, and enrichment of core functional components.

[0009] To address the aforementioned technical problems, this invention provides a method for preparing a compound probiotic Cistanche deserticola fermented beverage that tonifies the kidneys and nourishes the liver, comprising the following steps:

[0010] S1. Mix and sterilize Cistanche deserticola, Poria cocos, malt, functional ingredients and natural selenium-strontium mineral water to obtain fermentation substrate;

[0011] The functional components are xylooligosaccharides, d-ribose, disodium pyrroloquinoline quinone, allulose, monk fruit extract, xanthan gum, pectin, and sodium bicarbonate.

[0012] S2. Activate the strains of Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus rhamnosus, and Lactobacillus plantarum, respectively, centrifuge and freeze-dry them, and then mix them to obtain a compound probiotic powder; culture the compound probiotic powder in MRS medium to obtain a compound probiotic solution;

[0013] S3. The compound probiotic liquid is inoculated into the fermentation substrate for fermentation, and after sterilization, the compound probiotic Cistanche deserticola fermented drink for tonifying the kidney and nourishing the liver is obtained.

[0014] This invention utilizes a specific ratio of fermentation substrate, a compound probiotic fermentation system, and a fermentation process to prepare a high-performance compound probiotic Cistanche deserticola fermented beverage that nourishes the kidneys and liver. It efficiently converts and enriches the core functional components of Cistanche deserticola, such as echinacoside, verbascoside, and β-(1-3)-glucan from Poria cocos. Through compound probiotic fermentation, a natural acidity is added, significantly improving the flavor and palatability of the product. Systematic optimization of process parameters ensures the product has a high viable count and stable functional quality.

[0015] Furthermore, in S1, the weight parts of each component in the fermentation substrate are as follows: 5 parts of Cistanche deserticola, 4-6 parts of Poria cocos, 8-12 parts of malt, 2.5-3 parts of xylooligosaccharide, 1.5-2 parts of d-ribose, 0.002-0.01 parts of disodium pyrroloquinoline quinone, 10-15 parts of allulose, 0.5-2 parts of Siraitia grosvenorii extract, 0.05-0.1 parts of xanthan gum, 0.1-0.2 parts of pectin, 0.4-0.6 parts of sodium bicarbonate, and 140-160 parts of natural selenium-strontium mineral water.

[0016] Furthermore, in S1, the selenium content in the natural selenium-strontium mineral water is 0.01-0.05 mg / L, and the strontium content is 0.24-0.52 mg / L.

[0017] Furthermore, in S2, the *Lactobacillus casei* is CICC 6114, *Lactobacillus acidophilus* is CICC 6087, *Lactobacillus rhamnosus* is CICC 6133, and *Lactobacillus plantarum* is CICC 21790.

[0018] Furthermore, in S2, the activation conditions for the strain are: temperature 35-40℃, 12-24h.

[0019] Furthermore, in S2, the viable count of the compound probiotic solution is >1×10⁻⁶. -8 CFU / mL.

[0020] Furthermore, in S2, the mass ratio of Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus rhamnosus, and Lactobacillus plantarum in the compound probiotic powder is 1:(0.8-1.2):(0.8-1.2):(0.8-1.2).

[0021] Furthermore, in S3, the inoculation amount of the compound probiotic liquid is 1-3% of the fermentation substrate volume.

[0022] Furthermore, in S3, the fermentation conditions are: temperature 20-40℃, rotation speed 30-100r / min, and time 24-72h.

[0023] The second aspect of this invention provides a compound probiotic Cistanche deserticola fermented drink prepared by the preparation method described in the first aspect, which tonifies the kidney and nourishes the liver.

[0024] The third aspect of this invention provides the application of the compound probiotic Cistanche fermented beverage described in the second aspect in kidney-tonifying and liver-nourishing products.

[0025] The beneficial effects of this invention are:

[0026] This invention utilizes an innovative integrated formula technology combining Cistanche deserticola, malt, and Poria cocos as food and medicine ingredients, multifunctional factor compounding, and probiotic fermentation to prepare a kidney-tonifying probiotic Cistanche deserticola fermented beverage with high active ingredients, clear efficacy, and excellent flavor. The probiotic fermentation process of this invention has a clear effect on the targeted transformation, core component enrichment, and deep biomodification of Cistanche deserticola, Poria cocos, and malt. The product also possesses highly active ingredients (echinacoside, verbascoside, Poria cocos β-(1-3)-glucan), abundant probiotic metabolites (short-chain fatty acids), and optimized product characteristics (low pH, high antioxidant). Its technological advantages and clear functional orientation of "tonifying the liver and kidneys, and nourishing essence and blood" demonstrate its excellent development potential and application value. Specifically:

[0027] This invention utilizes precise fermentation with compound probiotics to achieve efficient conversion and specific enrichment of core functional components in Cistanche deserticola, including echinacoside, verbascoside, and β-(1-3)-glucan from Poria cocos. Echinacoside concentrations reached 0.138 mg / mL, and verbascoside concentrations reached 0.087 mg / mL. The fermentation process significantly enhanced the target components while also increasing the total flavonoid and total phenolic content, achieving a targeted enhancement of the "tonifying liver and kidneys, nourishing essence and blood" effects and a systematic improvement in the overall level of active substances.

[0028] During the fermentation process of this invention, the probiotics exhibit active metabolism, resulting in a significant increase in the content of short-chain fatty acids: acetic acid reaches 15.13 μg / mL, propionic acid reaches 5.52 μg / mL, butyric acid reaches 4.54 μg / mL, and isobutyric acid reaches 5.27 μg / mL. These metabolites can effectively regulate the intestinal flora and acid-base environment, forming a synergistic effect with the nourishing effects of raw materials such as Cistanche deserticola, malt, and Poria cocos, providing modern microecological support for the traditional "liver and kidney tonification" function.

[0029] The probiotic metabolism of this invention significantly reduces the product's pH value from 5.39 to 3.39, creating a natural acidic environment that helps inhibit unwanted bacteria and enhances storage stability. Simultaneously, in vitro antioxidant capacity is enhanced, with increased DPPH free radical scavenging rate, hydroxyl free radical scavenging rate, and superoxide anion free radical scavenging rate. Furthermore, the flavor compounds produced during fermentation effectively improve the taste of the raw materials, increasing product acceptability.

[0030] This invention's fermentation process successfully transforms Cistanche deserticola raw material into a functional system rich in target active ingredients and probiotic metabolites. The extremely significant increase in short-chain fatty acids and the efficient enrichment of characteristic active ingredients jointly demonstrate that the compound probiotic formulation used has a highly specific biocatalytic ability and conversion efficiency for this raw material system.

[0031] This invention uses Cistanche deserticola as its core ingredient, combined with Poria cocos and malt to form a synergistic formula that tonifies the kidneys, strengthens the spleen and stomach, and promotes qi circulation and relieves bloating. It breaks through the limitations of traditional single-ingredient tonic formulas, achieving the effects of aiding digestion, strengthening the spleen, and tonifying without causing stagnation. It also incorporates multifunctional factors such as xylooligosaccharides, D-ribose, and disodium pyrroloquinoline quinone, as well as a healthy sugar system including D-allulose and monk fruit, constructing an integrated system that provides energy replenishment, strong antioxidant properties, mitochondrial protection, prebiotic proliferation, and optimized taste. Attached Figure Description

[0032] To more clearly illustrate the technical solution of the present invention, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0033] Figure 1This is a comparison chart of the active ingredients of the fermented beverage obtained under the optimal conditions of Example 2 and Comparative Example 1 of the present invention;

[0034] Figure 2 This is a comparison chart of the antioxidant capacity of the fermented beverage obtained under the optimal conditions of Example 2 and Comparative Example 1 of the present invention;

[0035] Figure 3 This is a comparison chart of the fatty acid content of fermented beverages obtained under the optimal conditions of Example 2 and Comparative Example 1 of the present invention. Detailed Implementation

[0036] The technical solution of the present invention will be clearly and completely described below with reference to specific embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0037] In this embodiment of the invention, *Lactobacillus casei* CICC 6114, *Lactobacillus acidophilus* CICC 6087, *Lactobacillus rhamnosus* CICC 6133, and *Lactobacillus plantarum* CICC 21790 were all purchased from the China Industrial Microbial Culture Collection Center (CICC).

[0038] MRS medium: 10g peptone, 10g beef extract, 5g yeast extract, 1g Tween-80, 2g dipotassium hydrogen phosphate, 5g sodium acetate, 2g triammonium citrate, 0.2g magnesium sulfate, 0.05g manganese sulfate, bring to a final volume of 1L.

[0039] Example 1

[0040] This embodiment relates to a method for preparing a compound probiotic Cistanche deserticola fermented beverage that tonifies the kidneys and nourishes the liver, comprising the following steps:

[0041] (1) Preparation of medicinal and edible substrate: Weigh 5g of Cistanche deserticola, 5g of Poria cocos, 10g of malt, 2.8g of xylooligosaccharide, 1.8g of d-ribose, 5mg of disodium pyrroloquinoline quinone, 1g of Siraitia grosvenorii, 0.069g of xanthan gum, 0.1104g of pectin, 12g of D-allulose, and 0.5g of sodium bicarbonate and mix them evenly. Add 150mL of natural selenium-strontium mineral water (selenium content is 0.01-0.05mg / L, strontium content is 0.24-0.52mg / L) and stir evenly to obtain a mixed substrate. Transfer the mixed substrate to a sterilizer and sterilize it with high pressure steam at 121℃ for 15 minutes. After cooling, the fermentation substrate is obtained.

[0042] (2) Lactobacillus casei CICC 6114, Lactobacillus acidophilus CICC 6087, Lactobacillus rhamnosus CICC 6133, and Lactobacillus plantarum CICC 21790 were inoculated into sterilized 250 mL Erlenmeyer flasks containing 50 mL of MRS medium, and incubated at 37 °C for 24 h. The resulting bacterial solutions were centrifuged at 5000 r / min for 20 min, and the precipitate was dried using a freeze dryer to prepare a viable count of 1.0 × 10⁻⁶. 9 Each strain of probiotic powder was prepared at CFU / g. The probiotic powders of *Lactobacillus casei* CICC 6114, *Lactobacillus acidophilus* CICC 6087, *Lactobacillus rhamnosus* CICC 6133, and *Lactobacillus plantarum* CICC 21790 were mixed in a 1:1:1:1 ratio to obtain a compound probiotic powder.

[0043] (3) Take the compound probiotic powder and inoculate it into a sterilized 250mL Erlenmeyer flask containing 50mL MRS medium. Then, seal the Erlenmeyer flask and place it in a shaker at 37℃ for static incubation until the logarithmic growth phase. Centrifuge at 4℃ and 4000rpm for 10 minutes, discard the supernatant, wash with sterile water, and prepare a bacterial suspension (bacterial concentration >1×10⁻⁶). -8 (CFU / mL).

[0044] (4) Add the bacterial suspension to the fermentation substrate at an inoculation rate of 1% (volume ratio), and ferment at a constant temperature of 30°C for 72 hours with stirring at a speed of 50 rpm to obtain the fermentation broth.

[0045] (5) Post-processing: After fermentation, shake the fermentation liquid to mix well, take 5mL for dilution and spread for later use. After fermentation, transfer to a sterile bottle for pasteurization, sterilize at 80℃ for 30min, and store at 4℃ for later use.

[0046] Example 2: Optimization of Fermentation Conditions

[0047] This embodiment optimizes the fermentation process by referring to the steps in Embodiment 1:

[0048] (1) Orthogonal experimental design: The process conditions affecting the fermentation capacity of the compound strain were selected as factors for the orthogonal experiment. Simultaneously, the levels were determined, and an appropriate orthogonal table was selected for the header design. The factors required in the experiment were arranged in each column of the orthogonal table (Table 1). Based on the design principles of the orthogonal table and the determined factors and levels, the orthogonal table was designed using SPSS. L9(3) 3 An orthogonal array was used, with each factor having 3 levels. The core evaluation index was "viable cell count (CFU / mL) after fermentation". The results were repeated 3 times and the average value was taken. The specific design and data are as follows:

[0049] Table 1

[0050]

[0051] (2) Orthogonal results analysis: The orthogonal results and range analysis are shown in Tables 2 and 3. The key factors affecting the viable cell count are ranked as follows: inoculum size (B) > fermentation time (A) > stirring speed (C) > fermentation temperature (D). Among them, the inoculum size is the primary control factor. The optimal fermentation process combination is A4B3C4D1, that is, fermentation time 72h, inoculum size 3.0%, stirring speed 100rpm, and fermentation temperature 37℃; the viable cell count is lowest when dissolved oxygen is insufficient at 0rpm, and highest when dissolved oxygen is moderate at 100rpm.

[0052] Table 2

[0053]

[0054] Table 3

[0055]

[0056] As can be seen from Tables 2 and 3, the conditions of this invention are met when the volume ratio of the inoculum is 1-3%, the temperature is 20-40℃, the rotation speed is 30-100r / min, and the fermentation time is 24-72h.

[0057] (3) Fermentation verification

[0058] Three batches of samples were prepared repeatedly according to the optimal process (A4B3C4D1), and the viable count, active ingredients, and stability indicators were tested to verify the process repeatability. The viable count was 7.5 × 10⁻⁶. 9 CFU / mL, 7.3×10 9 CFU / mL, 7.6×10 9 The CFU / mL data is stable, proving that the optimized parameters are reliable.

[0059] Comparative Example 1

[0060] This comparative example is similar to the optimal fermentation conditions (A4B3C4D1) in Example 2, except that the step of preparing the compound probiotic suspension is omitted, and the compound probiotic suspension in the fermentation process is replaced with the same volume of sterilized water.

[0061] Test case

[0062] The antioxidant capacity of the product obtained under the optimal conditions (A4B3C4D1) in Example 2 and the product prepared in Comparative Example 1 were tested, and the contents of polysaccharides and total phenolic flavonoids were determined. The specific methods are as follows:

[0063] (1) Superoxide anion free radical scavenging ability

[0064] Take 4.5 mL of 0.05 mol / L pH 8.2 Tris-HCl buffer solution and preheat it in a 25°C water bath for 20 min. Add 1 mL of fermentation broth and 0.4 mL of 25 mmol / L pyrogallol solution, mix well, and react in a 25°C water bath for 5 min. Terminate the reaction by adding 1.0 mL of 8 mol / L HCl. Using Tris-HCl buffer solution as a reference, measure the absorbance at 299 nm and calculate the scavenging rate. The blank control group is represented by 1 mL of sample solvent instead of the sample. The formula for calculating the scavenging rate is: Superoxide anion radical scavenging rate (%) = [(A0-(A1-A2) / A0]×100%, where A0 is the absorbance of the blank control solution; A1 is the absorbance of the sample test tube; and A2 is the absorbance of the sample background tube.

[0065] (2) DPPH free radical scavenging ability

[0066] Add 40 μL of fermentation broth to 4 mL of 0.1 mmol / L DPPH-methanol solution, then add 450 μL of 50 mmol / L Tris-HCl buffer (7.4), and incubate at 25 °C for 30 min. Use deionized water as a reference solution and measure the absorbance at 517 nm. DPPH free radical scavenging rate (%) = [(A0-(A1-A2) / A0] × 100%, where A0 is the absorbance of the blank control solution; A1 is the absorbance of the sample test tube; and A2 is the absorbance of the sample background tube.

[0067] (3) Hydroxyl radical scavenging ability

[0068] Add 135 μL of fermentation broth to 1.4 mL of 6 mmol / L H₂O₂, then add 0.6 mL of 20 mmol / L sodium salicylate and 2 mL of 1.5 mmol / L ferrous sulfate. Incubate at 37 °C for 1 h. Use deionized water as the reference solution. Measure the absorbance at 562 nm. Hydroxyl radical scavenging capacity (%) = [(A0 - (A1 - A2) / A0] × 100%, where A0 is the absorbance of the blank control solution; A1 is the absorbance of the sample test tube; and A2 is the absorbance of the sample background tube.

[0069] (4) Polysaccharide determination (phenol-sulfuric acid method):

[0070] Sample preparation: Add 5 times the volume of anhydrous ethanol to the fermentation broth and let it stand overnight at 4°C. Filter the mixture and dry the precipitate at 60°C to constant weight to obtain crude polysaccharide.

[0071] Construction of the standard curve: Prepare 5% phenol solution and 100 mg / L glucose standard solution. Accurately pipette 0, 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, and 1.0 mL of glucose standard solution into test tubes, and bring the volume to 2.0 mL with pure water. Add 1.0 mL of phenol solution and 5 mL of concentrated sulfuric acid solution. React for 10 min, then mix well and react at 30℃ for 20 min. Zero the instrument with a blank and measure the absorbance at 490 nm. Plot the glucose concentration on the x-axis and the absorbance on the y-axis to construct the standard curve.

[0072] Sample determination: Take 0.01g of crude polysaccharide and dilute to 50mL. The determination method is the same as that of the standard curve.

[0073] (5) Total phenols

[0074] Construction of the standard curve: Total phenols were tested using the Folin-Ciocalteu method. First, a pyrogallol standard solution was prepared. 5 mg of pyrogallol was weighed and diluted to 25 mL with methanol, yielding a concentration of 0.2 mg / mL. Accurately pipette 0, 0.01 mL, 0.02 mL, 0.05 mL, 0.1 mL, 0.12 mL, and 0.15 mL of pyrogallol solution into test tubes. The volume was brought to 0.5 mL with ethanol, and then 0.5 mL of Folin-Ciocalteu solution was added. The mixture was allowed to stand for 3 minutes, followed by 1 mL of 15% sodium carbonate solution. The mixture was then allowed to stand for 30 minutes. After standing, the mixture was centrifuged at 3500 rpm for 3 minutes. Using a reagent blank as a control, the absorbance was measured at 760 nm. A standard curve was plotted with absorbance (A) on the ordinate and pyrogallol concentration on the ordinate.

[0075] (6) Total flavonoids

[0076] Construction of the standard curve: The sodium nitrite-aluminum nitrate colorimetric method was used. First, a rutin standard solution was prepared by weighing 10.4 mg of rutin and diluting it to 25 mL with 80% ethanol. Accurately pipette 0.025 mL, 0.06 mL, 0.12 mL, 0.24 mL, and 0.48 mL of rutin solution, add ethanol to make up to 0.5 mL of reagent, mix well, add 0.15 mL of 5% sodium nitrite, and let stand for 6 min. After standing, add 0.15 mL of 10% aluminum nitrate and let stand for 6 min. Add 2 mL of 4% sodium hydroxide solution and dilute to 5 mL with water, shake to mix, and let stand for 3 min. Measure the absorbance at 508 nm. Plot the absorbance (A) on the ordinate and the rutin concentration on the abscissa to construct the standard curve.

[0077] (7) Echinacoside

[0078] Echinacoside was detected by high performance liquid chromatography using a C18 column at 35℃, a mobile phase flow rate of 1.0 ml / min, an injection volume of 5 μL, and a detection wavelength of UV-330 nm. Gradient elution was performed in a solvent-acetonitrile system containing 0.05% phosphoric acid at 5%-100% acetonitrile from 0 to 10.0 min, and at 10.0-13.0 min, 100% acetonitrile was obtained.

[0079] (8) Verbascoside

[0080] Verbascoside was detected by high performance liquid chromatography using a C18 column at 35℃, a mobile phase flow rate of 1.0 ml / min, an injection volume of 5 μL, and a detection wavelength of UV-330 nm. The mobile phase gradient elution conditions were 10%→40% acetonitrile in an aqueous solution containing 0.05% phosphoric acid, with a run time of 30 min.

[0081] The optimal fermentation conditions (A4B3C4D1) of Example 2 and the test results of Comparative Example 1 are shown in Table 4. Figures 1-3 As shown.

[0082] Table 4

[0083]

[0084] From Table 4 and Figures 1-3 As shown, fermentation of medicinal herbs with mixed probiotics can improve various indicators:

[0085] The content of core active ingredients has been significantly increased: echinacoside reached 0.138 mg / mL (an increase of 34%), and verbascoside reached 0.087 mg / mL (an increase of 34%). While significantly enhancing the target components, the fermentation process also increased the total flavonoid content by 96% and the total phenolic content by 86%, achieving a targeted enhancement of the "liver and kidney tonifying, essence and blood nourishing" effects and a systematic improvement in the overall level of active substances.

[0086] During fermentation, probiotics exhibited active metabolism, resulting in a significant increase in the content of short-chain fatty acids: acetic acid reached 15.13 μg / mL (an increase of 41%), propionic acid reached 5.52 μg / mL (an increase of 159%), butyric acid reached 4.54 μg / mL (an increase of 344%), and isobutyric acid reached 5.27 μg / mL (an increase of 75%). These metabolites can effectively regulate the intestinal flora and pH environment, synergizing with the nourishing effects of ingredients such as Cistanche deserticola, malt, and Poria cocos, providing modern microecological support for the traditional "liver and kidney tonification" function.

[0087] The metabolism of probiotics significantly reduced the product's pH value from 5.39 to 3.39, creating a natural acidic environment that helps inhibit unwanted bacteria and enhances storage stability. Simultaneously, the in vitro antioxidant capacity was also enhanced, with DPPH free radical scavenging rate increasing by 40.7%, hydroxyl free radical scavenging rate by 26.3%, and superoxide anion free radical scavenging rate by 7.3%.

[0088] The present invention has been described in detail above with reference to specific embodiments and exemplary examples; however, these descriptions should not be construed as limiting the present invention. Those skilled in the art will understand that various equivalent substitutions, modifications, or improvements can be made to the technical solutions and embodiments of the present invention without departing from the spirit and scope of the invention, and all such modifications and improvements fall within the scope of the present invention. The scope of protection of the present invention is defined by the appended claims.

Claims

1. A preparation method of a kidney-tonifying and liver-nourishing compound probiotic Cistanche Fermented Drink, characterized in that, Includes the following steps: S1. Mix and sterilize Cistanche deserticola, Poria cocos, malt, functional ingredients and natural selenium-strontium mineral water to obtain a fermentation substrate; The functional components are xylooligosaccharides, d-ribose, disodium pyrroloquinoline quinone, allulose, monk fruit extract, xanthan gum, pectin, and sodium bicarbonate. S2. Activate the strains of Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus rhamnosus, and Lactobacillus plantarum, respectively, centrifuge and freeze-dry them, and then mix them to obtain a compound probiotic powder; culture the compound probiotic powder in MRS medium to obtain a compound probiotic solution; S3. The compound probiotic liquid is inoculated into the fermentation substrate for fermentation, and after sterilization, the compound probiotic Cistanche deserticola fermented drink for tonifying the kidney and nourishing the liver is obtained.

2. The preparation method of the kidney-tonifying and liver-nourishing compound probiotic cistanche fermented drink according to claim 1, characterized in that, In S1, the weight parts of each component in the fermentation substrate are as follows: 5 parts of Cistanche deserticola, 4-6 parts of Poria cocos, 8-12 parts of malt, 2.5-3 parts of xylooligosaccharide, 1.5-2 parts of d-ribose, 0.002-0.01 parts of disodium pyrroloquinoline quinone, 10-15 parts of allulose, 0.5-2 parts of Siraitia grosvenorii extract, 0.05-0.1 parts of xanthan gum, 0.1-0.2 parts of pectin, 0.4-0.6 parts of sodium bicarbonate, and 140-160 parts of natural selenium-strontium mineral water.

3. The preparation method of the compound probiotic Cistanche fermented beverage for tonifying the kidney and nourishing the liver as described in claim 1, characterized in that, In S1, the natural selenium-strontium mineral water contains 0.01-0.05 mg / L of selenium and 0.24-0.52 mg / L of strontium.

4. The preparation method of the compound probiotic Cistanche deserticola fermented beverage for tonifying the kidney and nourishing the liver as described in claim 1, characterized in that, In S2, the activation conditions for the strain are: temperature 35-40℃, 12-24h.

5. The preparation method of the compound probiotic Cistanche fermented beverage for tonifying the kidney and nourishing the liver as described in claim 1, characterized in that, In S2, the viable cell count of the complex probiotic bacteria liquid is >1x10 -8 CFU / mL.

6. The preparation method of the compound probiotic Cistanche fermented beverage for tonifying the kidney and nourishing the liver as described in claim 1, characterized in that, In S2, the mass ratio of Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus rhamnosus, and Lactobacillus plantarum in the compound probiotic powder is 1:(0.8-1.2):(0.8-1.2):(0.8-1.2).

7. The preparation method of the compound probiotic Cistanche fermented beverage for tonifying the kidney and nourishing the liver as described in claim 1, characterized in that, In S3, the inoculation amount of the compound probiotic liquid is 1-3% of the fermentation substrate volume.

8. The preparation method of the compound probiotic Cistanche fermented beverage for tonifying the kidney and nourishing the liver as described in claim 1, characterized in that, In S3, the fermentation conditions are: temperature 20-40℃, rotation speed 30-100r / min, and time 24-72h.

9. A compound probiotic Cistanche deserticola fermented drink for tonifying the kidney and nourishing the liver, prepared by the preparation method according to any one of claims 1-8.

10. The application of the compound probiotic Cistanche fermented beverage according to claim 9 in a kidney-tonifying and liver-nourishing product.

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