Kidney function health wine based on traditional Chinese medicine fermentation and preparation method thereof

Abstract

The invention relates to the technical field of health care products, and discloses kidney function health wine based on traditional Chinese medicine fermentation and a preparation method of the kidney function health wine. 6-10 parts of cistanche deserticola; 9-30 parts of radix astragali and 6-12 parts of fructus lycii; the fermentation strain comprises lactobacillus acidophilus, lactobacillus plantarum, bifidobacterium longum subsp. Longum, saccharomyces cerevisiae and bacillus subtilis, and the fermentation strain comprises the following strains: lactobacillus acidophilus JYLC-374, lactobacillus plantarum JYLC-482, bifidobacterium longum subsp. Longum JYBB-163, saccharomyces cerevisiae JYSC-710 and bacillus subtilis JYBS-312. By adopting the technical scheme that a traditional Chinese medicine compound formula is combined with specific probiotic fermentation, the technical effects of enhancing the bioavailability of active ingredients and promoting kidney function regulation are achieved, and the dissolution rate and the biotransformation rate of herbal medicine active substances are improved.

Description

Technical Field

[0001] The invention relates to the technical field of health care products, in particular to a kidney function health-preserving wine based on traditional Chinese medicine fermentation and a preparation method thereof. Background Art

[0002] The improvement and protection of renal function has been a topic of great concern in the medical field in recent years. As an important detoxification organ of the human body, the kidney undertakes the important functions of removing metabolic waste and regulating the internal environment. However, due to unhealthy lifestyles, eating habits, environmental pollution and other factors, more and more people are facing the problem of renal function decline, especially the prevalence of chronic kidney disease (CKD) has been rising year by year. Existing treatment methods are mostly single drug interventions, such as the use of vasodilators, diuretics and other drugs.

[0003] At present, many studies focus on the application of botanicals and natural ingredients to explore their potential in kidney protection. Botanicals have become the main direction of modern Chinese medicine for the treatment of kidney diseases because of their natural, safe and few side effects. For example, common plants such as astragalus, wolfberry, and Cistanche have been proven to have certain kidney protective effects and can play a health-care role by improving kidney blood circulation, regulating immune response, etc. The therapeutic effect of a single plant ingredient is usually limited, and the active ingredients of botanicals are easily affected by factors such as the environment and extraction process, resulting in unstable effects and difficulty in meeting the needs of clinical treatment.

[0004] As active ingredients that can regulate intestinal microecology, probiotics have a significant effect on improving intestinal health and enhancing the immune system. Intestinal health is closely related to kidney health. Intestinal microbial imbalance may lead to the occurrence and development of chronic kidney disease. Through the auxiliary role of probiotics, the intestinal environment can be improved and toxin accumulation can be reduced, thereby indirectly having a positive impact on kidney health. At present, the application of most probiotics is still limited to its direct impact on the gastrointestinal tract. Few studies have combined probiotics with traditional botanicals to exert multiple effects to enhance kidney function. Summary of the invention

[0005] In view of the deficiencies in the prior art, the present invention provides a kidney function health wine based on Chinese medicine fermentation and a preparation method thereof, which solves the problems of low utilization rate of ingredients and single function in traditional Chinese medicine wine.

[0006] To achieve the above purpose, the present invention is implemented by the following technical scheme: a kidney function health wine based on traditional Chinese medicine fermentation, comprising the following ingredients:

[0007] 3-9 portions of ginseng;

[0008] 6-10 portions of Cistanche deserticola;

[0009] Astragalus 9-30 parts;

[0010] 6-12 servings of wolfberry;

[0011] The fermentation bacteria are 1-5 g / L, and the fermentation bacteria include Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium longum subspecies longum, Saccharomyces cerevisiae and Bacillus subtilis.

[0012] Furthermore, ginseng (red ginseng) contains ginsenosides, which can enhance immunity and antioxidant capacity; the phenylethanol glycoside components of Cistanche deserticola have the effect of nourishing the kidney and replenishing essence; astragalus polysaccharides and wolfberry polysaccharides synergistically regulate kidney metabolism; betaine and flavonoids in wolfberry assist in anti-inflammatory and protect kidney cells, Lactobacillus acidophilus and Lactobacillus plantarum lower the pH value of the fermentation liquid by producing acid, inhibiting the growth of miscellaneous bacteria; Bifidobacterium longum releases short-chain fatty acids (SCFAs) to promote the dissolution of active ingredients in traditional Chinese medicine; Saccharomyces cerevisiae metabolizes to produce ethanol and esters to enhance the flavor of the wine; Bacillus subtilis secretes cellulase to decompose the cell walls of medicinal materials and release the effective ingredients in the cells.

[0013] Preferably, the fermentation bacteria include the following bacteria: Lactobacillus acidophilus JYLC-374, Lactobacillus plantarum JYLC-482, Bifidobacterium longum subspecies longum JYBB-163, Saccharomyces cerevisiae JYSC-710, and Bacillus subtilis JYBS-312.

[0014] Preferably, the mixing ratio of the fermentation bacteria is: Lactobacillus acidophilus 0.2-0.5 g / L, Lactobacillus plantarum 0.2-0.5 g / L, Bifidobacterium longum subsp. longum 0.2-0.5 g / L, Saccharomyces cerevisiae 0.2-0.5 g / L, and Bacillus subtilis 0.2-0.5 g / L.

[0015] Preferably, the fermentation bacteria are activated before use, the activation conditions being a temperature of 30 to 37° C. for 12 to 24 hours using sterile physiological saline.

[0016] Furthermore, the activation treatment of the fermentation bacteria restarts the metabolic activity of dormant bacteria in a temperature environment of 30 to 37°C, promotes the recovery of cell membrane fluidity and key enzyme systems, and guides the bacterial community to complete the transition from damage repair to exponential proliferation within 12 to 24 hours; the osmotic pressure protection and ion balance of sterile saline (0.85% to 0.9% NaCl) not only avoids osmotic stress but also induces the bacteria to consume energy storage substances to stimulate metabolic potential, ultimately achieving directional activation of highly active and highly stress-resistant fermentation bacteria.

[0017] The preparation method of kidney function health wine comprises the following steps:

[0018] First, select the herbs ginseng, cistanche, astragalus, and wolfberry, and process them separately. Ginseng is processed into red ginseng, cistanche and astragalus are cut into small pieces, and wolfberry is cleaned and set aside;

[0019] Add ginseng, cistanche, astragalus and wolfberry into distilled water according to the formula ratio and soak for 12 to 24 hours at a temperature of 20 to 30°C;

[0020] The soaked herbal liquid is boiled for extraction at a temperature of 80-95°C for 2-3 hours to ensure that the active ingredients in the herbs can be fully released, and then the boiled liquid is filtered to remove impurities;

[0021] After the filtered herbal liquid is cooled to 30-40°C, the fermentation bacteria are added, and the amount of the fermentation bacteria is 1-5g / L;

[0022] The mixed liquid is put into a fermentation container and fermented under temperature control conditions. The fermentation temperature is controlled at 25-35°C and the fermentation time is 5-10 days.

[0023] Furthermore, during the activation phase of the strains, metabolic activity is restored through temperature and time control to ensure rapid proliferation of the bacterial flora in the initial stage of fermentation; segmented temperature control (25-30°C) allows different strains of bacteria to function in the optimal metabolic window period (such as yeast extending the fermentation cycle at low temperatures to improve the efficiency of ester production).

[0024] Preferably, the ginseng is processed into red ginseng, and fresh ginseng is washed, steamed and dried, the steaming temperature is 100-120°C, the duration is 2-4 hours, the drying temperature is 50-60°C, and the ginseng is dried to a moisture content of ≤10%.

[0025] Preferably, the cleaning step uses an ultrasonic cleaning machine for 10 to 20 minutes to remove impurities on the ginseng surface, the steaming step uses a steam sterilizer to ensure that the ginseng is mature, and the drying step uses a circulating hot air drying oven.

[0026] Preferably, the fermentation temperature is controlled at 25-30° C. and the fermentation time is 5-8 days to ensure the best fermentation effect. The pH value and alcohol concentration of the fermentation liquid are checked every 2 days.

[0027] Preferably, the wine needs to be treated by activated carbon adsorption and membrane filtration, and the pore size of the filter membrane is 0.2 to 0.45 μm to remove impurities.

[0028] Furthermore, activated carbon adsorption removes phenolic impurities, and membrane filtration intercepts microbial residues to ensure the clarity of the wine; the dynamic balance of esterification reaction and redox reaction during aging (such as the complexation of ginsenosides and ethanol) can enhance the stability of the active ingredients.

[0029] Preferably, the filtered wine needs to be aged for 15 to 30 days at a temperature of 10 to 25° C., and the wine is stored in a light-proof, sealed container.

[0030] The present invention provides a kidney function health wine based on Chinese medicine fermentation and a preparation method thereof. It has the following beneficial effects:

[0031] 1. The present invention adopts a technical solution of combining a traditional Chinese medicine compound formula with fermentation of specific probiotics to achieve the technical effect of enhancing the bioavailability of active ingredients and promoting renal function regulation. Compared with the solution of soaking or extracting a single herb in the prior art, the present invention improves the dissolution rate and bioconversion rate of herbal active substances through fermentation with Lactobacillus acidophilus, Lactobacillus plantarum, etc., and solves the problem of low utilization rate and single function of ingredients in traditional Chinese medicine wine.

[0032] 2. The present invention uses sterile physiological saline to pretreat the strains, achieving the technical effect of improving the survival rate of the strains and optimizing the fermentation process. Compared with the problem of insufficient activity caused by poor adaptability of the bacteria in the prior art of directly adding strains for fermentation, the present invention improves the initial activity of the bacteria through the activation step, making the fermentation start more efficient and solving the problems of low competitiveness of the bacteria and delayed fermentation.

[0033] 3. The present invention adopts a dual optimization strategy of low-temperature decoction extraction and slow cooling followed by inoculation and fermentation of the strains, achieving the technical effect of fully releasing the herbal medicinal ingredients and protecting the activity of the strains. Compared with the high-temperature direct extraction or rapid cooling methods in the prior art, the present invention precisely controls the temperature so that the effective ingredients are not destroyed during the high-temperature extraction process, while avoiding damage to the strains caused by sudden temperature changes, thereby solving the problems of reduced efficacy and low survival rate of the strains.

[0034] 4. The present invention adopts activated carbon adsorption combined with membrane filtration and aging treatment to achieve the technical effect of removing impurities, improving taste and product stability. Compared with the simple filtration or short-term storage solution in the prior art, the present invention ensures the purity of the wine through double purification technology and optimizes the flavor through low-temperature aging, solving the problem of rough taste and easy degradation of ingredients in traditional Chinese medicinal wine. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 The present invention is a flow chart of the method. DETAILED DESCRIPTION

[0036] The following will be combined with the drawings in the specification of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0037] Please refer to the attached Figure 1 :

[0038] Example 1

[0039] Raw material ratio

[0040] 6 portions of ginseng (red ginseng)

[0041] 8 servings of Cistanche deserticola

[0042] 15 servings of Astragalus

[0043] 9 servings of wolfberry

[0044] 60 parts of distilled water

[0045] Fermentation bacteria (Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium longum subsp. longum, Saccharomyces cerevisiae, Bacillus subtilis mixed, 1:1:1:1:1) 2g / L

[0046] Preparation steps

[0047] Herbal pretreatment

[0048] Wash the ginseng, steam sterilize it at 120℃ for 3 hours, and dry it with circulating hot air at 50℃ until the moisture content is less than 10%.

[0049] Slice Cistanche and Astragalus, wash wolfberry, remove impurities by ultrasonic for 15 minutes, drain and set aside.

[0050] Active ingredient extraction

[0051] Soak all medicinal materials in distilled water according to the formula for 20 hours at 28°C.

[0052] Slowly boil and extract at 80℃ for 2.5 hours, filter to remove the residue and retain the clear liquid.

[0053] Bacteria activation

[0054] The fermentation bacteria (2 g / L) were placed in 0.88% NaCl sterile saline and activated at 30°C for 18 hours.

[0055] Fermentation

[0056] The herbal extract is cooled to 35°C, activated bacteria are added, and placed in a fermentation tank.

[0057] Fermentation was carried out at a constant temperature of 30°C for 6 days, and the pH value was monitored every 2 days and stabilized at 4.2-4.5.

[0058] Purification and aging

[0059] Filter (0.2μm filter membrane) and remove impurities by activated carbon adsorption.

[0060] Aged at 15℃ for 20 days, sealed and protected from light.

[0061] Example 2

[0062] Raw material ratio

[0063] 4 portions of ginseng (red ginseng)

[0064] 10 servings of Cistanche deserticola

[0065] 20 servings of Astragalus

[0066] 6 servings of wolfberry

[0067] 58 parts of distilled water

[0068] Fermentation bacteria (Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium longum subsp. longum, Saccharomyces cerevisiae, and Bacillus subtilis mixed, 1:1:1:1:1) 3g / L.

[0069] Preparation steps

[0070] Chinese medicine treatment

[0071] The red ginseng was steamed at 110°C for 2.5 hours and air-dried at 50°C, with a final moisture content of 8%.

[0072] Slice the Astragalus and Cistanche, and wash the wolfberries with cold water.

[0073] extract

[0074] Soak at 22℃ for 12 hours.

[0075] Heat and boil at 90℃ for 2 hours, filter and keep the liquid.

[0076] Pre-activation of bacteria

[0077] Use 0.9% NaCl solution and activate at 36°C for 24 hours.

[0078] Fermentation

[0079] After cooling to 35℃, add the bacteria and ferment at 28℃ for 8 days.

[0080] The pH was checked every 48 hours and ultimately maintained at 4.0-4.3.

[0081] Post-processing

[0082] Activated carbon adsorption method removes impurities.

[0083] Aging at low temperature (12℃) for 30 days and stored in sealed container.

[0084] Example 3

[0085] Raw material ratio

[0086] 9 portions of ginseng (red ginseng)

[0087] 6 servings of Cistanche deserticola

[0088] 25 servings of Astragalus

[0089] 12 servings of wolfberry

[0090] 48 parts distilled water

[0091] Fermentation bacteria (Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium longum subsp. longum, Saccharomyces cerevisiae, and Bacillus subtilis mixed, 1:1:1:1:1) 5g / L.

[0092] Preparation steps

[0093] Chinese medicine treatment

[0094] Steam red ginseng at 120℃ for 2 hours and dry at 55℃ until the moisture content is ≤9%.

[0095] Cut the astragalus into sections and wash the wolfberries at low temperature (18°C).

[0096] Soaking and extraction

[0097] Soak in distilled water at 25°C for 24 hours.

[0098] Heat and boil at 85℃ for 3 hours and filter.

[0099] Bacteria activation

[0100] Activated with 0.85% NaCl saline and cultured at 30°C for 12 hours.

[0101] Fermentation

[0102] The extract was cooled to 32°C and inoculated with fermentation bacteria.

[0103] After 7 days of fermentation at 26°C, the pH dropped to 4.1-4.4.

[0104] Purification and storage

[0105] Filter through a 0.22 μm filter membrane to remove the precipitate.

[0106] Aging at 18℃ for 15 days, storage at low temperature.

[0107] Example 4

[0108] Raw material ratio

[0109] 5 portions of ginseng (red ginseng)

[0110] 9 servings of Cistanche deserticola

[0111] 30 servings of Astragalus

[0112] 6 servings of wolfberry

[0113] 50 parts distilled water

[0114] Fermentation bacteria (Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium longum subsp. longum, Saccharomyces cerevisiae, and Bacillus subtilis mixed, 1:1:1:1:1) 4g / L.

[0115] Preparation steps

[0116] Herbal treatment

[0117] Steam red ginseng at 105℃ for 3 hours, soak wolfberry in cold water for 10 minutes, and slice astragalus and Cistanche.

[0118] extract

[0119] Soak in 30℃ water for 16 hours and boil at 85℃ for 2 hours.

[0120] Bacteria culture

[0121] 0.9% NaCl solution, activated at 37°C for 24 hours.

[0122] Fermentation

[0123] Ferment at 28°C for 5 days, test pH every day, and finally control it at 4.0-4.2.

[0124] Post-processing

[0125] After filtration and activated carbon adsorption treatment, it was aged at 12℃ for 25 days.

[0126] Example 5

[0127] Raw material ratio

[0128] 3 portions of ginseng (red ginseng)

[0129] 7 portions of Cistanche deserticola

[0130] 12 servings of Astragalus

[0131] 10 servings of wolfberry

[0132] 63 parts of distilled water

[0133] Fermentation bacteria (Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium longum subsp. longum, Saccharomyces cerevisiae, and Bacillus subtilis mixed, 1:1:1:1:1) 2g / L.

[0134] Preparation steps

[0135] Pre-treatment

[0136] Steam red ginseng at 110℃ for 2 hours, slice astragalus, and wash wolfberry at low temperature for 15 minutes.

[0137] extract

[0138] Soak in 25℃ water for 18 hours, boil at 90℃ for 2.5 hours, and filter.

[0139] Bacteria activation

[0140] 0.85% NaCl saline, activated at 30℃ for 16 hours.

[0141] Fermentation

[0142] Fermentation was carried out at 27°C for 6 days and the pH was stabilized at 4.2-4.5.

[0143] Purification and storage

[0144] Filtered with 0.22μm membrane and aged at 16℃ for 20 days.

[0145] Comparative Example 1 (corresponding to Example 1, without using fermentation bacteria)

[0146] Herbal pretreatment

[0147] In the same manner as in Example 1, ginseng, Cistanche deserticola, Astragalus, and Lycium barbarum were selected.

[0148] Ginseng was steam sterilized at 120℃ for 3 hours and dried with circulating hot air at 50℃ until the moisture content was less than 10%.

[0149] Slice Cistanche and Astragalus, wash wolfberry, use ultrasonic cleaning machine to remove impurities for 15 minutes, drain and set aside.

[0150] Active ingredient extraction

[0151] Put the medicinal materials into distilled water according to the formula ratio, and soak them at 28℃ for 20 hours to allow the ingredients to fully dissolve.

[0152] Decoction was performed at 80°C for 2.5 hours with appropriate stirring during the process to increase the release rate of the active ingredients.

[0153] The residue was removed by filtration to obtain a clear solution, which was then cooled to 35°C.

[0154] No fermentation bacteria, natural fermentation

[0155] The filtered medicinal solution is directly poured into the fermentation tank without adding any exogenous fermentation bacteria, and is sealed and left to stand.

[0156] It is placed at 30℃ for 6 days and fermented by relying on the natural flora in the environment without any external regulation.

[0157] Purification and aging

[0158] The solution was filtered through a 0.2 μm filter membrane to remove most of the precipitate and suspended matter.

[0159] Without activated carbon adsorption treatment, it was directly aged at 15℃ for 20 days and stored away from light.

[0160] Comparative Example 2 (corresponding to Example 2, without bacterial activation treatment)

[0161] Chinese medicine pretreatment

[0162] Select red ginseng that meets the standards, steam it at 110℃ for 2.5 hours, and dry it at 50℃ until the moisture content drops to 8%.

[0163] Slice Cistanche and Astragalus, and wash wolfberry with cold water and set aside.

[0164] Active ingredient extraction

[0165] Let it soak at 22℃ for 12 hours to allow the herbal ingredients to slowly dissolve.

[0166] Heat and boil at 90℃ for 2 hours and stir evenly to improve the efficiency of ingredient release.

[0167] Filter and remove the residue, retain the liquid and cool to 35°C.

[0168] Unactivated bacteria are added directly

[0169] 3 g / L of mixed fermentation bacteria (Lactobacillus acidophilus, Lactobacillus plantarum, etc.) were directly added to the cooled extract without activation by 0.9% NaCl solution and without temperature control.

[0170] After cooling to 35°C, it was fermented directly and placed at 28°C for 8 days without the activation stage and adaptation culture.

[0171] Post-processing

[0172] The precipitate was removed by filtration using a 0.2 μm filter membrane.

[0173] After activated carbon adsorption treatment, it is aged at 12°C for 30 days and stored in a sealed container.

[0174] Comparative Example 3 (corresponding to Example 3, with different extraction temperature and time)

[0175] Chinese medicine pretreatment

[0176] Select qualified red ginseng, steam it at 120℃ for 2 hours, and dry it at 55℃ until the moisture content is ≤9%.

[0177] Cut the astragalus into sections and wash the wolfberries at low temperature (18°C) to ensure there are no impurities.

[0178] Low temperature extraction (extended time)

[0179] Soak at 30℃ for 24 hours to allow the herbal ingredients to precipitate naturally.

[0180] Low-temperature boiling at 60°C for 6 hours was used instead of extraction at 85°C for 3 hours to avoid high temperature destruction of some volatile components, but it was not verified whether it affected the extraction efficiency.

[0181] Filter and remove the residue to obtain the extract.

[0182] Bacteria activation

[0183] Activated with 0.85% NaCl saline and cultured at 30°C for 12 hours.

[0184] Fermentation

[0185] After cooling to 32℃, add 5g / L fermentation bacteria.

[0186] Fermentation was carried out at 26°C for 7 days, during which the pH value was monitored every 48 hours, and the temperature was not adjusted.

[0187] Purification and storage

[0188] Filter with a 0.22 μm filter membrane to remove the precipitate.

[0189] Aging at 18℃ for 15 days, storage at low temperature.

[0190] Comparative Example 4 (corresponding to Example 4, without activated carbon adsorption purification)

[0191] Herbal treatment

[0192] The same method as in Example 4 was used to process red ginseng, Cistanche deserticola, Astragalus, and Lycium barbarum, including the steps of steaming, cutting, and washing.

[0193] extract

[0194] Soak in 30℃ water for 16 hours to dissolve the herbal ingredients.

[0195] Decoction at 85℃ for 2 hours to fully release the active ingredients, and filter out the residue.

[0196] Bacteria culture

[0197] 0.9% NaCl solution, activated at 37℃ for 24 hours to restore the strain to the best state.

[0198] Fermentation

[0199] Ferment at 28°C for 5 days, test pH every day, and finally control it at 4.0-4.2.

[0200] Post-treatment (without activated carbon adsorption)

[0201] Direct filtration with a 0.22 μm membrane was performed without the use of activated carbon adsorption, and potential precipitates and impurities were not removed.

[0202] Aged at 12℃ for 25 days, stored in sealed container away from light.

[0203] Comparative Example 5 (corresponding to Example 5, without aging treatment)

[0204] Pre-treatment

[0205] Select high-quality red ginseng, steam it at 110℃ for 2 hours, slice astragalus, and wash wolfberry at low temperature for 15 minutes.

[0206] Carry out herbal pretreatment according to specifications to ensure the stable quality of herbal medicine.

[0207] Active ingredient extraction

[0208] Soak at 25℃ for 18 hours to allow the herbal ingredients to be evenly released.

[0209] Decoction at 90℃ for 2.5 hours, filter to remove the residue and ensure the liquid is clear.

[0210] Bacteria activation

[0211] The cells were activated with 0.85% NaCl saline and cultured at 30°C for 16 hours to improve the survival rate of the cells.

[0212] Fermentation

[0213] Fermentation was carried out at 27°C for 6 days, and the pH value was strictly monitored, and finally stabilized at 4.2-4.5.

[0214] Post-processing (unaged)

[0215] Use 0.22μm filter membrane to remove impurities and suspended matter.

[0216] The aging step is omitted and the wine is directly bottled and sealed. There is no low-temperature storage process, and no flavor enhancement or ingredient stability optimization is performed.

[0217] Experiment 1: Verify the effect of fermentation strains on product activity

[0218] Experimental procedures

[0219] Sample preparation

[0220] Samples were prepared according to the processes of Example 1 and Comparative Example 1 to ensure that the raw materials and operations were consistent, with the only variable being whether fermentation bacteria were added.

[0221] For Example 1, Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium longum subsp. longum, Saccharomyces cerevisiae, and Bacillus subtilis were added according to the formula ratio, 2 g / L, and inoculated after a total of 18 hours of activation.

[0222] For Comparative Example 1, no exogenous bacteria were added, and only microorganisms in the natural environment were relied upon for self-fermentation.

[0223] Fermentation conditions

[0224] Fermentation temperature: 30℃, fermentation time: 6 days.

[0225] Samples were taken every 2 days to measure pH value and count bacteria.

[0226] The final product was taken to determine the content of active ingredients and metabolites, including flavonoids, polysaccharides, saponins, etc. (HPLC), organic acid content (HPLC-MS), flavor substances (electronic nose), and bacterial flora structure (16S rRNA sequencing).

[0227] Testing and data recording

[0228] The total flavonoid content (mg / mL) was determined by ultraviolet-visible spectrophotometry (UV-Vis).

[0229] The total polysaccharide content (mg / mL) was determined by the phenol-sulfuric acid method.

[0230] The saponin content was analyzed by high performance liquid chromatography (HPLC).

[0231] GC-MS was used to detect flavor substances and record the changes in the peak area of ​​volatile components. 16S rRNA sequencing was used to analyze the bacterial composition and the proportion of dominant bacterial species.

[0232] Three parallel groups were set up for each experiment, and the mean was taken for statistical analysis.

[0233] Experimental data

[0234] Table 1. Effects of fermentation strains on active ingredients and metabolites

[0235]

[0236]

[0237] Experimental Summary

[0238] The addition of fermentation strains significantly increased the accumulation of metabolites, especially the concentrations of lactic acid and acetic acid in the fermentation system. This change promoted the decrease in pH value and formed a microenvironment more conducive to enzymatic reactions. Due to the lack of the leading role of exogenous flora, the acidity of the final product of Comparative Example 1 was maintained at a high level, and the lactic acid production was significantly low. This shows that in the early stage of fermentation, exogenous bacteria can quickly occupy the ecological niche and enter the stage of vigorous metabolism, while the adaptability of natural flora is weak, and the metabolic efficiency is far less than that of the screened and optimized composite bacterial system.

[0239] From the changes in active ingredients such as polysaccharides, flavonoids, and saponins, the role of the fermented bacteria is not limited to metabolic acid production, but also promotes the degradation of plant cell walls to a certain extent, releasing more active ingredients. In particular, the polysaccharide content in Example 1 is much higher than that in Comparative Example 1, which may be related to the decomposition of cellulose and hemicellulose by the fermented bacteria. In contrast, in the natural fermentation environment, the enzyme activity of the flora is limited and the degree of cell wall degradation is low, so the release efficiency of the active ingredients is not as good as that of the system with added fermented bacteria.

[0240] The comparative analysis of flavor substances revealed the sensory differences under different fermentation modes. The peak area of ​​the main flavor substances in Example 1 is relatively high, which means that the fermentation bacteria can effectively mediate the synthesis of volatile compounds. The moderate increase in ethanol content also reflects the fermentation contribution of brewer's yeast. In Comparative Example 1, the accumulation of flavor substances is relatively slow, and the richness of the overall aroma is reduced. This phenomenon can be attributed to the limited metabolic capacity of natural flora and the lack of strong biocatalysis, which results in the final product being not full in flavor level and presenting a relatively single flavor characteristic.

[0241] Experiment 2: Verify the effect of bacterial activation on fermentation effect

[0242] Experimental procedures

[0243] Sample preparation

[0244] Example 2 (bacteria activated with 0.9% NaCl solution for 24 hours before inoculation) and Comparative Example 2 (direct inoculation of unactivated bacteria) were used, with the only variable being whether bacteria activation was performed.

[0245] The same formula was used in both groups to ensure consistent fermentation substrate.

[0246] Fermentation conditions

[0247] Fermentation was carried out at a constant temperature of 28°C for 8 days, and samples were taken every 48 hours to detect pH value, bacterial growth, and metabolite content.

[0248] The initial bacterial inoculation volume was 3 g / L to ensure consistent bacterial counts.

[0249] Testing items and methods

[0250] Bacterial growth curve (OD600 measured by spectrophotometer).

[0251] Number of viable bacteria (CFU count by dilution plating method).

[0252] Lactic acid, acetic acid and other organic acid content (HPLC detection).

[0253] pH value changes (measured by acidometer).

[0254] Alcohol content of the final product (GC-MS detection).

[0255] Experimental setup

[0256] Three parallel experiments were performed, each group was repeated 3 times, and the mean was taken for data analysis.

[0257] Experimental data

[0258] Table 2. Effect of strain activation on the fermentation process

[0259]

[0260]

[0261] Experimental Summary

[0262] The activation of strains has a decisive influence on the start-up speed and final yield of fermentation. From the initial growth data, the bacterial proliferation of Example 2 is more rapid, and the OD600 value on the second day of fermentation is significantly higher than that of Comparative Example 2. The physiological adaptation period of the unactivated strains is longer, and the proliferation rate is significantly delayed, which indicates that the unactivated strains need to readjust the cell membrane osmotic pressure and metabolic state when adapting to the matrix environment, resulting in slow initial growth. On the contrary, the activated bacterial flora has completed physiological adaptation in advance, and quickly enters the logarithmic growth phase after inoculation, occupying a metabolic advantage.

[0263] The accumulation trend of organic acids further illustrates the improvement of metabolic efficiency by strain activation. In Example 2, the content of lactic acid and acetic acid is about twice that of Comparative Example 2, and the pH value decreases faster. This shows that the activated bacterial flora can quickly metabolize substrates, produce lactic acid and acetic acid, and inhibit the formation of unnecessary by-products. Due to the weak metabolism of the unactivated strains in the initial stage, the acid accumulation rate is slow, and the pH value does not decrease significantly. This low-acid environment may weaken the antibacterial ability of the fermentation broth, increase the competitive pressure of miscellaneous bacteria, and further affect the quality and flavor of the final product.

[0264] The change in alcohol content also reflects the difference in fermentation efficiency. The final ethanol content of Example 2 reached 0.55%, which is more than double that of Comparative Example 2. This may be related to the degree of activation of Saccharomyces cerevisiae. After entering the fermentation system, the bacterial flora adapted to physiological saline can quickly restore the activity of the ethanol fermentation pathway. When the unactivated strains adapt to the new environment, part of the energy is used for cell repair, resulting in obstruction of alcohol metabolism and a reduction in final yield. This phenomenon once again shows that strain activation not only affects the proliferation rate, but also determines the distribution of metabolic flux to a certain extent, thereby affecting the quality of the final product.

[0265] Experiment 3: Verify the effect of extraction temperature and time on active ingredients

[0266] Experimental procedures

[0267] Sample preparation

[0268] Using Example 3 (extraction at 85°C for 3 hours) and Comparative Example 3 (extraction at 60°C for 6 hours), the only variables were the extraction temperature and time.

[0269] The same extraction material and extraction solvent (70% ethanol) were used to ensure consistency of other conditions.

[0270] After pretreatment, extraction was performed at different temperatures and times.

[0271] Extraction conditions

[0272] For Example 3, the extraction temperature was set to 85°C and the extraction time was 3 hours.

[0273] For Comparative Example 3, the extraction temperature was set to 60° C. and the extraction time was 6 hours.

[0274] Using an ultrasonic oscillation extractor, 200 mL of solvent was taken for each sample, the ultrasonic frequency was 40 kHz, and the power was 200 W.

[0275] Testing items and methods

[0276] Active ingredient content (flavonoids, polysaccharides, saponins): analyzed by high performance liquid chromatography (HPLC).

[0277] Antioxidant capacity (DPPH free radical scavenging rate assay).

[0278] Volatile flavor compounds: gas chromatography-mass spectrometry (GC-MS) was used.

[0279] Total acidity: measured by an acidometer.

[0280] Sediment content: Determined by filtration and weight method.

[0281] Experimental setup

[0282] Three parallel experiments were set up in each group to ensure that each experiment was repeated three times, and the mean was calculated and statistical analysis was performed.

[0283] Experimental data

[0284] Table 3. Effects of extraction temperature and time on active ingredients and metabolites

[0285]

[0286]

[0287] Experimental Summary

[0288] The extraction temperature and time have a significant effect on the dissolution efficiency of the active ingredients in the plant. From the data, the extraction effect of Example 3 (85°C, 3 hours) is significantly better than that of Comparative Example 3 (60°C, 6 hours). The higher extraction temperature accelerates the interaction between the solvent and the sample, causing more flavonoids, polysaccharides and other components to be dissolved and enter the extract. This phenomenon may be related to the effect of temperature on cell walls and cell membranes. High temperature destroys the structure of plant tissues, thereby releasing more active compounds. On the contrary, although the lower extraction temperature is milder, the extraction rate is slower, and the content of active ingredients in the final product is significantly reduced.

[0289] In terms of antioxidant capacity, the antioxidant activity of Example 3 is significantly higher than that of Comparative Example 3. High temperature accelerates the dissolution of phenolic substances, especially flavonoids, which have a strong ability to scavenge free radicals. Therefore, a higher extraction temperature not only increases the concentration of active ingredients, but also increases the antioxidant capacity of the extract. Lower temperatures may lead to incomplete dissolution of certain antioxidant components, so that their activity cannot be fully demonstrated.

[0290] In the analysis of volatile components, the peak area of ​​the aroma substance in Example 3 is larger, which may be related to the accelerated release rate of volatile compounds under high temperature conditions. In contrast, many volatile substances may be well retained during low-temperature extraction, resulting in a lower peak area. Therefore, high-temperature extraction not only helps to increase the dissolution rate of active ingredients, but also promotes the release of flavor substances. This shows that in the process of flavor optimization and component extraction, high-temperature extraction can obtain more target products in a shorter time.

[0291] Experiment 4: Verify the effect of activated carbon adsorption purification on product purity and flavor

[0292] Experimental procedures

[0293] Sample preparation

[0294] Example 4 (using activated carbon adsorption treatment) and Comparative Example 4 (not using activated carbon adsorption treatment) were used, and the only variable was whether activated carbon adsorption was performed.

[0295] The same fermentation product was treated under the same conditions, activated carbon powder was added respectively, and the adsorption time was 24 hours (Example 4). Comparative Example 4 did not add any adsorbent.

[0296] The dosage of activated carbon was 5 g / L, and the adsorption treatment was carried out using the immersion method.

[0297] Adsorption treatment

[0298] After 24 hours, the two groups of samples were filtered to remove the activated carbon to obtain the purified fermentation broth.

[0299] The activated carbon and its adsorbed impurities are separated using conventional filtration devices (paper filters).

[0300] Testing items and methods

[0301] Wine turbidity (determined by turbidity meter).

[0302] Precipitation formation (store at constant temperature for 30 days to observe the precipitation).

[0303] Content of bitter substances (such as phenols) (UV-Vis spectroscopy analysis).

[0304] Flavor analysis (GC-MS detection of volatile components).

[0305] Experimental setup

[0306] Three parallel experiments were set up in each group, repeated three times, and the mean was calculated for data analysis.

[0307] Experimental data

[0308]

[0309] Experimental Summary

[0310] The use of activated carbon adsorption purification significantly improved the quality of the fermented product, especially in terms of wine clarity and sediment removal. In Example 4, the wine turbidity was significantly lower than that in Comparative Example 4, indicating that activated carbon had a significant effect on removing suspended matter and impurities in the wine. The reduction in sediment further demonstrated the adsorption capacity of activated carbon on impurities, especially the removal of phenolic compounds that may affect the clarity of the wine. In contrast, the untreated Comparative Example 4 still had a high turbidity and the formation of sediment was also more obvious, reflecting the poor stability of the treated wine.

[0311] The analysis of flavor substances showed that the peak area of ​​the main flavor substances in Example 4 was larger, indicating that the activated carbon not only removed unnecessary impurities, but also promoted the release of flavor substances to some extent. In contrast, in Comparative Example 4, which did not use activated carbon, the release of flavor substances was relatively delayed and the flavor complexity was insufficient. The significant reduction in bitterness (Example 4 scored significantly lower than Comparative Example 4) also shows that the activated carbon effectively removed phenolic substances that may cause bitterness, which helps to improve the taste of the wine and the balance of flavor.

[0312] In general, activated carbon adsorption not only improves the clarity of the wine, but also removes unnecessary impurities and bitter substances. This process may be related to the physical adsorption and chemical adsorption mechanism of activated carbon. Activated carbon adsorbs most of the impurities such as phenols and fatty acids through its huge specific surface area and pore structure, while providing a purer basis for the flavor substances in the wine.

[0313] Experiment 5: Verify the effect of aging on the flavor and stability of wine components

[0314] Experimental procedures

[0315] Sample preparation

[0316] Using Example 5 (aging for 20 days) and Comparative Example 5 (no aging), the only variable was whether aging was performed.

[0317] The samples in Example 5 were placed in a constant temperature room at 20° C. and aged for 20 days. Comparative Example 5 was not aged, and the samples were tested immediately after the experiment.

[0318] Aging process

[0319] During the aging period, check the changes in the wine regularly and record the temperature and humidity data.

[0320] During the aging process, no external ingredients or regulators are added, ensuring the process occurs naturally.

[0321] Testing items and methods

[0322] Flavor substance analysis: GC-MS is used to detect changes in volatile components, mainly analyzing changes in aroma substances.

[0323] Wine acidity and alcohol content: Acidity is measured by titration and alcohol content is analyzed using GC.

[0324] Microbiological stability: The microbial content in the wine is analyzed through total bacteria count and yeast tests.

[0325] Total antioxidant capacity: The antioxidant activity was tested using the DPPH free radical scavenging method.

[0326] Experimental setup

[0327] Three parallel experiments were set up for each group to ensure that each experiment was repeated three times, and the mean was taken for statistical analysis, and the data were summarized for further comparison.

[0328] Experimental data

[0329] Table 6. Effects of aging on wine flavor, acidity, alcohol content and antioxidant capacity

[0330]

[0331] Experimental Summary

[0332] Through aging treatment, the flavor substances and antioxidant activity of the wine are significantly improved. The peak area of ​​flavor substances in Example 5 is much higher than that in Comparative Example 5, especially in the accumulation of volatile aroma components. After 20 days of aging, the aromatic substances in the wine are released and stabilized, which may be related to the formation of ester substances and the redox reaction of polyphenols. The increase of these flavor substances directly enhances the complexity and layering of the wine. In contrast, the flavor substances in Comparative Example 5 are relatively single, and the peak area is significantly lower than that in Example 5.

[0333] Acidity and alcohol content also showed differences. The esterification reaction during the aging process may have reduced the acidity to a certain extent, reflecting the optimization effect of aging on the balance of the wine. The acidity of the wine in Example 5 was relatively low (1.92mL NaOH), which may be due to the slow conversion of acidic components over time, reducing the overall sourness. At the same time, the alcohol content remained basically stable, indicating that aging had no significant effect on the stability of alcohol.

[0334] Microbiological analysis also showed that after 20 days of aging, the number of microorganisms in Example 5 was relatively low, indicating that the aging process helped to stabilize and inhibit the natural microorganisms in the wine. Comparative Example 5 showed a higher number of microorganisms, which may be due to the lack of microbial inhibition during the aging process, resulting in the continued growth of certain bacteria or yeasts. Overall, aging not only improves the flavor and antioxidant capacity, but also effectively improves the sensory quality of the wine and reduces possible microbial risks.

[0335] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A kidney function health wine based on Chinese medicine fermentation, characterized in that: Includes the following ingredients: 3-9 portions of ginseng; 6-10 portions of Cistanche deserticola; Astragalus 9-30 parts; 6-12 servings of wolfberry; The fermentation bacteria are 1-5 g / L, and the fermentation bacteria include Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium longum subspecies longum, Saccharomyces cerevisiae and Bacillus subtilis.

2. The kidney function health wine based on traditional Chinese medicine fermentation according to claim 1, characterized in that: The fermentation strains include the following strains: Lactobacillus acidophilus JYLC-374, Lactobacillus plantarum JYLC-482, Bifidobacterium longum subspecies longum JYBB-163, Saccharomyces cerevisiae JYSC-710, and Bacillus subtilis JYBS-312.

3. The kidney function health wine based on Chinese medicine fermentation according to claim 1, characterized in that: The mixing ratio of the fermentation bacteria is: 0.2-0.5 g / L of Lactobacillus acidophilus, 0.2-0.5 g / L of Lactobacillus plantarum, 0.2-0.5 g / L of Bifidobacterium longum subsp. longum, 0.2-0.5 g / L of Saccharomyces cerevisiae, and 0.2-0.5 g / L of Bacillus subtilis.

4. The kidney function health wine based on Chinese medicine fermentation according to claim 1, characterized in that: The fermentation bacteria are activated before use, the activation conditions being a temperature of 30 to 37° C. for 12 to 24 hours, and the activation is performed using sterile physiological saline.

5. A method for preparing a kidney function health wine, which is applied to the kidney function health wine based on Chinese medicine fermentation according to claims 1-4, characterized in that: The following steps are involved: First, select the herbs ginseng, cistanche, astragalus, and wolfberry, and process them separately. Ginseng is processed into red ginseng, cistanche and astragalus are cut into small pieces, and wolfberry is cleaned and set aside; Add ginseng, cistanche, astragalus and wolfberry into distilled water according to the formula ratio and soak for 12 to 24 hours at a temperature of 20 to 30°C; The soaked herbal liquid is boiled for extraction at a temperature of 80-95°C for 2-3 hours to ensure that the active ingredients in the herbs can be fully released, and then the boiled liquid is filtered to remove impurities; After the filtered herbal liquid is cooled to 30-40°C, the fermentation bacteria are added, and the amount of the fermentation bacteria is 1-5g / L; The mixed liquid is put into a fermentation container and fermented under temperature control conditions. The fermentation temperature is controlled at 25-35°C and the fermentation time is 5-10 days.

6. The method for preparing the kidney function health wine according to claim 5, characterized in that: The ginseng is processed into red ginseng. Fresh ginseng is washed, steamed and dried. The steaming temperature is 100-120° C. for 2-4 hours. The drying temperature is 50-60° C. and the ginseng is dried until the moisture content is ≤10%.

7. The method for preparing the kidney function health wine according to claim 5, characterized in that: The cleaning step uses an ultrasonic cleaning machine for 10 to 20 minutes to remove impurities on the surface of the ginseng. The steaming step uses a steam sterilizer to ensure that the ginseng is mature. The drying step uses a circulating hot air drying oven.

8. The method for preparing the kidney function health wine according to claim 5, characterized in that: The fermentation temperature is controlled at 25-30°C and the fermentation time is 5-8 days to ensure the best fermentation effect. The pH value and alcohol concentration of the fermentation liquid are checked every 2 days.

9. The method for preparing the kidney function health wine according to claim 5, characterized in that: The wine needs to be treated by activated carbon adsorption and membrane filtration, and the pore size of the filter membrane is 0.2-0.45 μm to remove impurities.

10. The method for preparing the kidney function health wine according to claim 5, characterized in that: The filtered wine needs to be aged for 15 to 30 days at a temperature of 10 to 25°C, and the wine should be stored in a light-proof, sealed container.

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