Probiotic fermented traditional Chinese medicine preparation as well as preparation method and application thereof

Abstract

The invention provides a probiotic fermented traditional Chinese medicine preparation as well as a preparation method and application thereof. The probiotic fermented traditional Chinese medicine preparation is prepared by taking a water extract of a traditional Chinese medicine composition as a fermentation substrate and fermenting by utilizing probiotics, the traditional Chinese medicine composition comprises coix seeds, poria cocos, Chinese waxgourd seeds, chrysanthemum, selfheal, black beans, mung beans and liquorice. The probiotics comprise lactobacillus casei, lactobacillus reuteri, bifidobacterium animalis subsp. Lactis and lactobacillus rhamnosus; the probiotics comprise lactobacillus casei, lactobacillus reuteri, bifidobacterium animalis subsp. Lactis and lactobacillus rhamnosus; the traditional Chinese medicine composition has the effects of clearing damp, promoting diuresis, tonifying spleen, stopping diarrhea, clearing away heat and toxic materials, clearing liver and purging fire through combination of the medicines, so that the effect of removing damp toxin accumulation type stains is achieved; the traditional Chinese medicine raw material extracting solution is fermented through specific probiotics, macromolecular substances are decomposed, the bioavailability of traditional Chinese medicine active ingredients is improved, the probiotic fermented traditional Chinese medicine preparation has a good treatment effect on damp toxin accumulation type spots, the defect that the spots are difficult to cure and persistent and stubborn damp toxin spots are overcome, and the recurrence rate is reduced.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of traditional Chinese medicine fermentation, and particularly relates to a probiotic fermented traditional Chinese medicine preparation and a preparation method and application thereof. BACKGROUND

[0002] There are mainly two sources of dampness in the human body: external dampness and internal dampness. If a person lives or works in a damp, dark, and poorly ventilated environment for a long time, external dampness will easily invade the human body; if a person often gets wet, wades in water, or does not dry the body and change dry clothes in time after swimming, external dampness will also invade the human body; and if a person directly blows cold wind, uses air conditioning, or takes a cold shower after intense exercise or a lot of sweating, the dampness will be trapped in the body. Traditional Chinese medicine believes that "the spleen governs the transformation of water and dampness", and the spleen is the "core organ" responsible for the digestion and absorption of food and water and the transformation of the food and water into fine substances for distribution to the whole body and the metabolism of excess water. If the spleen is weak and the transformation function is abnormal, the food and water taken in cannot be normally transformed and distributed, but will stagnate and accumulate, forming internal dampness. Therefore, "all dampness and swelling are related to the spleen". Excessive consumption of ice cream, iced beverages, and raw fruits and the like can damage the spleen yang and reduce the transformation ability of the spleen, leading to internal dampness; overeating of fatty and greasy food can increase the burden of the spleen and stomach and exceed the transformation ability of the spleen, causing dampness and turbidity; irregular diet and overeating can damage the function of the spleen and stomach and affect the transformation; lack of exercise can make the blood circulation sluggish and the function of the spleen and stomach weakened, and the dampness in the body difficult to be discharged outside the body through sweat and metabolism. Long-term thinking, excessive pressure, and mental tension can also affect the transformation function of the spleen and stomach. Dampness toxin is a pathological product produced by the imbalance of water and dampness metabolism in the body, and long-term stagnation can affect the blood circulation and cause problems such as pigment deposition, dampness toxin accumulation type of color spots, and dull skin.

[0003] The mechanism of dampness toxin causing spots involves multiple factors. Dampness has the characteristics of stagnation, easily blocks the meridians, and makes the blood unable to nourish the skin, so the metabolism of melanin is blocked. At the same time, dampness toxin often combines with heat to form dampness-heat, stimulates the activity of tyrosinase, and accelerates the generation of melanin. Clinically, the dampness toxin accumulation type of color spots is often dark yellow, mainly distributed around the nose and mouth, and shows dark spots like dust. It is accompanied by fatigue, poor appetite, abdominal distension after eating, dizziness and drowsiness, belching, loose and sticky stool, light color and small amount of menstruation, excessive leukorrhea, red tongue, yellow and greasy fur, and slippery and rapid pulse. Modern medicine believes that this is related to chronic inflammation and accumulation of free radicals, and the dampness toxin state may aggravate the oxidative stress reaction and induce abnormal skin pigmentation.

[0004] The wet toxin accumulation type of color spot is difficult to eradicate, has a significant negative impact on the physiological function and mental health of patients, and causes many life troubles. The incidence rate of the disease is continuously high and the recurrence rate is high. In addition, the pace of modern society is accelerating, environmental pollution is intensifying, and bad living habits are common, so the prevalence rate is showing an upward trend, and the number of patients is increasing. With the improvement of social and economic level and the improvement of life quality, consumers' demand for natural, safe and efficient depigmentation methods is increasing, so the depigmentation product of the wet toxin accumulation type of color spot has become one of the research and development directions of the medical field.

[0005] Current clinical treatment mainly relies on oral administration of traditional Chinese medicine or external treatment (such as mask, fumigation, bath, acupuncture), but these methods generally have low significant effect rate, insignificant effect, high recurrence rate, poor convenience, and improper operation may damage the skin or waste the essence. Especially noteworthy is that the existing treatment fails to fully combine the scientific principles of "internal regulation of human body function balance and promotion of metabolism" and "utilization of probiotics to reduce internal wet and turbid". Due to the lack of a systematic and perfect depigmentation efficacy research method and product efficiency evaluation system, the depigmentation drug market is mixed. If consumers choose improperly, they not only cannot depigment, but also may cause side effects. Therefore, a new product with the function of removing the wet toxin accumulation type of color spot becomes the preferred choice.

[0006] The current production method of traditional Chinese medicine oral liquid mainly includes the steps of extraction, filtration, preparation, filling, sterilization and packaging of traditional Chinese medicine decoction pieces. The extraction of traditional Chinese medicine decoction pieces generally adopts water extraction and alcohol precipitation method (water decoction and then ethanol precipitation of impurities), ultrasonic extraction, microwave extraction and other methods to extract effective components from traditional Chinese medicine decoction pieces. The water extraction and alcohol precipitation method will remove high molecular substances indiscriminately, resulting in the precipitation and filtration of water-soluble effective components such as polysaccharides and flavonoids, and the cost is relatively high; the ultrasonic extraction of traditional Chinese medicine effective components has the disadvantages of difficult power matching, easy degradation of heat-sensitive components, insufficient compatibility of compound, unstable extraction efficiency and serious noise pollution; the microwave extraction method has high scale cost, uneven extraction efficiency, limited solvent selection, insufficient product stability and maintenance difficulty. SUMMARY

[0007] In order to overcome the problems existing in the prior art, one of the purposes of the present application is to provide a probiotic fermented traditional Chinese medicine preparation. The second purpose of the present application is to provide a preparation method of the probiotic fermented traditional Chinese medicine preparation. The third purpose of the present application is to provide the application of the probiotic fermented traditional Chinese medicine preparation.

[0008] In order to achieve the above purposes, the technical scheme adopted by the present application is: The first aspect of the present application provides a probiotic fermented traditional Chinese medicine preparation, which is prepared by using a water extract of a traditional Chinese medicine composition as a fermentation substrate and utilizing probiotics for fermentation; The traditional Chinese medicine composition comprises coix seed, tuckahoe, wax gourd seed, chrysanthemum, spirea japonica, black bean, green bean and licorice; The probiotics comprise lactobacillus casei, lactobacillus reuteri, animal bifidobacterium lactis and lactobacillus rhamnosus.

[0009] The coix seed is dried heavy red rose petal.

[0010] The coix seed in the traditional Chinese medicine prescription of the present application can benefit water and remove dampness, and can invigorate the spleen and stop diarrhea; the tuckahoe can benefit water and remove dampness, and can invigorate the spleen and calm the heart; the two drugs are combined, and have the effects of benefiting water and removing dampness, invigorating the spleen and stopping diarrhea, and calming the heart, and are the monarch drug; the wax gourd seed can benefit water and remove swelling, and can clear heat and remove fire; the chrysanthemum can calm the liver and improve eyesight, and can clear heat and resolve toxicity; the spirea japonica can clear the liver and remove fire, and can disperse stagnation; the two are combined, and are the ministerial drug; the black bean can benefit water and resolve toxicity; the green bean can clear heat and resolve summerheat, and can resolve toxicity; and the licorice can moderate the properties of other drugs, and is the ministerial drug. The combination of the drugs can achieve the effects of benefiting water and removing dampness, invigorating the spleen and stopping diarrhea, clearing heat and resolving toxicity, and clearing the liver and removing fire, so as to remove dampness and remove stagnation type of color spots. The above components are essential components, all raw materials are qualified after detection, and meet the requirements of the current edition of Chinese Pharmacopoeia. These components have the effects of invigorating the spleen and drying dampness, benefiting water and removing dampness, clearing heat and resolving toxicity, and clearing the liver and removing fire in the theory of traditional Chinese medicine. Through the way of diet therapy, the active ingredients of the medicine-food homology traditional Chinese medicine act on the human body, and the dampness and stagnation type of color spots are removed through internal regulation.

[0011] The traditional Chinese medicine prescription of the present application has the characteristics of synergistic effect and reduced toxicity, can reduce drug toxicity and optimize molecular structure; can precisely regulate intestinal microecology, inhibit the reproduction of pathogenic bacteria, and repair intestinal mucosal barrier; can quickly take effect and maintain long-acting, the probiotic metabolites have long-acting effect, and have the characteristics of long-lasting release of small molecules of traditional Chinese medicine.

[0012] The formula of the present application ferments the traditional Chinese medicine raw materials through specific probiotics (such as lactobacillus casei, lactobacillus reuteri, animal bifidobacterium lactis, lactobacillus rhamnosus, etc.), decomposes macromolecular substances, improves the bioavailability of active ingredients of traditional Chinese medicine, generates new active ingredients, enhances the original efficacy or produces new physiological regulation function, regulates intestinal microecology, restores the core function of spleen transportation, regulates immunity and metabolism, and blocks the pathological cycle of dampness evil; the effect is mild, and will not cause allergic reaction or toxic side effect.

[0013] Specifically, the formula of the present application enhances the bioavailability and efficacy of traditional Chinese medicine with specific probiotics. Metabolic products of probiotics, such as organic acids (lactic acid, acetic acid), vitamins (B group, K), short-chain fatty acids, etc., are produced during fermentation. These substances have the effects of regulating intestinal flora, antioxidant, anti-inflammatory, enhancing immunity, etc., and synergize with active ingredients of traditional Chinese medicine, forming a "1+1>2" effect, and more comprehensively targeting multiple links of chloasma formation. Specific strains can convert large molecules and poorly absorbed ingredients in the formula into smaller molecules, higher activity or better water-soluble substances, significantly improving their biological activity and enhancing the efficacy of chloasma removal, antioxidant, anti-inflammatory, endocrine regulation, etc. The fermentation process can effectively mask or transform the bitter, astringent and unpleasant odor of traditional Chinese medicinal materials, producing a more acceptable sweet and sour flavor (derived from organic acids), significantly improving the patient's compliance. The metabolic products such as organic acids produced during fermentation can reduce the pH value of the product, forming an environment that is not conducive to the growth of miscellaneous bacteria, while the specific strains themselves occupy the ecological niche, helping to inhibit the contamination of miscellaneous bacteria, thereby improving the biological stability of the product.

[0014] Preferably, the traditional Chinese medicine composition comprises 10-30 parts of yiyiren, 10-30 parts of fuling, 5-20 parts of dongguazi, 5-20 parts of juhua, 5-15 parts of xiaakucao, 5-15 parts of heidou, 5-15 parts of lvdou, and 5-15 parts of gancao by mass fraction.

[0015] More preferably, the mass ratio of each traditional Chinese medicine raw material of the traditional Chinese medicine composition meets at least one of the following conditions: 1) The mass ratio of yiyiren to fuling is 1:(0.8-1.2); 2) The mass ratio of yiyiren to dongguazi is 1:(0.5-0.75); 3) The mass ratio of yiyiren to juhua is 1:(0.5-0.75); 4) The mass ratio of yiyiren to xiaakucao is 1:(0.4-0.6); 5) The mass ratio of yiyiren to heidou is 1:(0.4-0.6); 6) The mass ratio of yiyiren to lvdou is 1:(0.4-0.6); 7) The mass ratio of yiyiren to gancao is 1:(0.4-0.6).

[0016] Preferably, the probiotics comprise 0.5-1.5 parts of lactobacillus casei, 0.5-1.5 parts of lactobacillus reuteri, 0.5-1.5 parts of bifidobacterium animalis lactis, and 0.5-1.5 parts of lactobacillus rhamnosus by mass fraction.

[0017] More preferably, the probiotics comprise 0.8-1.2 parts of lactobacillus casei.

[0018] More preferably, the probiotic bacteria comprise 0.8-1.2 parts of Lactobacillus rhamnosus.

[0019] More preferably, the probiotic bacteria comprise 0.8-1.2 parts of Bifidobacterium animalis ssp. lactis.

[0020] More preferably, the probiotic bacteria comprise 0.8-1.2 parts of Lactobacillus rhamnosus.

[0021] Preferably, the mass ratio of the probiotic bacteria to the water extract of the traditional Chinese medicine composition is (1-2):100.

[0022] More preferably, the mass ratio of the probiotic bacteria to the water extract of the traditional Chinese medicine composition is (1.3-1.7):100.

[0023] Preferably, the mass ratio of the probiotic bacteria to the traditional Chinese medicine composition is (0.0001-0.1):100.

[0024] More preferably, the mass ratio of the probiotic bacteria to the traditional Chinese medicine composition is (0.0001-0.001):100.

[0025] Further preferably, the mass ratio of the probiotic bacteria to the traditional Chinese medicine composition is (0.0001-0.01):100.

[0026] Preferably, the viable bacterial count of Lactobacillus casei, Lactobacillus rhamnosus, Bifidobacterium animalis ssp. lactis and Lactobacillus rhamnosus is not less than 1×10 10 CFU / mL.

[0027] More preferably, the viable bacterial count of Lactobacillus casei, Lactobacillus rhamnosus, Bifidobacterium animalis ssp. lactis and Lactobacillus rhamnosus is not less than 1×10 11 CFU / mL.

[0028] Further preferably, the concentration is 1.0~1.5×10 11 cfu / mL.

[0029] The second aspect of the present application provides a preparation method of the probiotic bacteria fermented traditional Chinese medicine preparation of the first aspect, comprising the following steps: Mixing Coicis Semen, Poria cocos, Benincasa cerifera, Chrysanthemum, Prunella vulgaris, Black beans, Green beans and Licorice, and performing water extraction to obtain a water extract; placing the water extract and probiotic bacteria in a fermentation tank for fermentation to prepare the probiotic bacteria fermented traditional Chinese medicine preparation Preferably, the temperature of the water extraction is 90~100℃.

[0030] Preferably, the other process parameters of the water extraction are as follows: the solid-liquid weight ratio is 1:(6-10); the extraction times are 1-2 times; and the single extraction time is 1-2 h.

[0031] More preferably, the weight ratio of the liquid to the feed is 1:(7-9).

[0032] Preferably, the process further includes the following step: after water extraction, the water extract is centrifuged and purified to obtain a centrifuged liquid.

[0033] The centrifugal purification process can remove drug residue and larger impurity particles.

[0034] Preferably, the pressure inside the fermenter during the fermentation process is less than 0.15 MPa.

[0035] More preferably, the pressure inside the fermenter is 0.05-0.07 MPa.

[0036] Preferably, the fermentation temperature is 36-38°C.

[0037] Preferably, the fermentation time is 16-30 hours.

[0038] More preferably, the fermentation time is 20-24 hours.

[0039] The third aspect of the present invention provides the use of the probiotic fermented traditional Chinese medicine preparation described in the first aspect in the preparation of a medicine for treating damp-heat accumulation or its related symptoms.

[0040] The accumulation of dampness and toxins refers to a serious pathological condition in which dampness accumulates, congeals, and transforms into toxins in the body, lurking deep within a certain part of the body, leading to local or systemic symptoms such as redness, swelling, heat, pain, oozing of foul odors, swelling and lumps, and a stubborn and difficult-to-cure condition.

[0041] Preferably, the symptoms caused by damp-heat accumulation are damp-heat accumulation type pigmentation.

[0042] The beneficial effects of this invention are: This invention provides a probiotic-fermented traditional Chinese medicine preparation. The herbal composition used in this preparation works synergistically to promote diuresis and eliminate dampness, strengthen the spleen and stop diarrhea, clear heat and detoxify, and clear liver fire, thereby removing pigmentation caused by damp-heat accumulation. Through fermentation of the herbal extracts by specific probiotics, macromolecular substances are decomposed, enhancing the bioavailability of the active ingredients and resulting in the following effects: generating new active ingredients, enhancing existing efficacy or producing new physiological regulatory functions; regulating the intestinal microecology and restoring the core function of spleen function; regulating immunity and metabolism, and blocking the pathological cycle of dampness; and having a mild effect, without causing allergic reactions or toxic side effects. This probiotic-fermented traditional Chinese medicine preparation has a good therapeutic effect on pigmentation caused by damp-heat accumulation, overcoming the difficulty in curing persistent and stubborn damp-heat-induced pigmentation and reducing the recurrence rate. Attached Figure Description

[0043] Figure 1The preparation process of the probiotic fermented traditional Chinese medicine preparation. DETAILED DESCRIPTION

[0044] The content of the present application is further illustrated in detail by specific examples. In the following examples, the raw materials, unless otherwise specified, can be obtained from conventional commercial channels or prepared and isolated by simple synthesis; the processes, unless otherwise specified, are conventional processes in the art.

[0045] Example 1 The present example provides an oral liquid for removing dampness-toxicity stagnation type of color spots, the probiotic fermented traditional Chinese medicine preparation is prepared by using probiotics to ferment the water extract of a traditional Chinese medicine composition as a fermentation substrate; the traditional Chinese medicine composition is obtained by mixing extraction of 10 parts of yiyi ren, 10 parts of fuling, 6.7 parts of dongguazi, 6.7 parts of chrysanthemum, 5 parts of xiaocucao, 5 parts of heidou, 5 parts of lüdou, and 5 parts of gancao; the added mixed lactobacillus has a mass of 0.05% of the total mass of the above raw materials. The mixed lactobacillus includes tyrocoliformis, royi viscous lactobacillus, animal bifidobacterium lactis subspecies, and rhamnose lactobacillus in a mass ratio of 0.5:0.5:0.5:0.5 (strain source: Guangzhou Microorganism Institute Group Co., Ltd., same below); the concentration of tyrocoliformis, royi viscous lactobacillus, animal bifidobacterium lactis subspecies, and rhamnose lactobacillus is 1×10 11 cfu / ml. The steps of the production method are as follows: Figure 1 ① Extraction: cut 10 parts of yiyi ren, 10 parts of fuling, 6.7 parts of dongguazi, 6.7 parts of chrysanthemum, 5 parts of xiaocucao, 5 parts of heidou, 5 parts of lüdou, and 5 parts of gancao into small pieces, mix them evenly, and put them into a reflux extraction tank; soak them in 8 times the weight of pure water for 0.5 h, then perform reflux extraction; control the heating temperature at 95℃, and extract for 1 hour each time, extract for 2 times, and combine the reflux extraction liquid for standby.

[0046] ② Centrifugal purification: slowly introduce the extraction liquid obtained in ① into a three-legged centrifuge, centrifugal purify it at a specified speed (generally not more than 1800 revolutions per minute), remove the dregs and larger impurity particles through the shaft and centrifugal bag, and transfer the centrifugal liquid to the fermentation tank.

[0047] ​③ Fermentation: Under sterile conditions, inoculate the seed tank with probiotic bacteria at an inoculation ratio of 1.5 / 100 into the fermentation tank, adjust the tank pressure (0.15 MPa or less) and temperature (36-38°C), and ferment for 20-24 hours to ensure normal growth of the bacteria. Continuously monitor the temperature and pressure (0.05-0.07 MPa) in the tank, and adjust according to the needs of the bacteria to ensure optimal fermentation process. Regularly sample to detect the growth of bacteria and the concentration of fermented liquid mixed with Lactobacillus, and release the tank when the bacterial growth and product quality meet the standards.

[0048] ④ Preparation: Weigh the amount of fermented liquid obtained in step ③ (140 L) and the amount of auxiliary materials such as purified water (60 L) and sweetener steviol glycoside (100 g) according to the formula requirements, and perform a review. Add the weighed fermented liquid, purified water and other auxiliary materials into the mixing tank, and mix and stir thoroughly.

[0049] ⑤ Automatic filling: Send the prepared liquid from step ④ to the automatic filling machine for automatic filling. During the automatic filling process, the flow rate and pressure of each step need to be accurately controlled to ensure that the content of each bag is uniform and accurate. After filling the liquid into the aluminum foil bag, the machine will melt and seal the bag opening to ensure that each bag opening is tightly closed to prevent the product from being contaminated or oxidized.

[0050] ⑥ Pasteurization: Place the semi-finished product from step ⑤ in a water bath at 72-82°C for 10-20 minutes for pasteurization to kill any disease-causing, deteriorating or unwanted fermentation microorganisms. After pasteurization, cool the semi-finished product with cold water to 4-8°C in time to inhibit bacterial growth and extend the shelf life.

[0051] ⑦ Packaging: According to the brand image, market demand and the characteristics of the wet toxin accumulation type color spot oral liquid, select appropriate packaging materials and patterns to make cartons and cartons for packaging to enhance the readability and identifiability of the packaging. Thus, the finished product of the wet toxin accumulation type color spot oral liquid can be obtained.

[0052] Example 2 The present embodiment provides a wet toxin accumulation type color spot oral liquid, which is prepared by fermenting a traditional Chinese medicine composition using probiotics. The traditional Chinese medicine composition is extracted from 30 parts of yiyi, 30 parts of fuling, 20 parts of dongguazi, 20 parts of chrysanthemum, 15 parts of xiaokucao, 15 parts of heidou, 15 parts of lüdou, and 15 parts of gancao. The mass of the added mixed lactobacillus is 0.3% of the total mass of the above raw materials. The mixed lactobacillus includes Lactobacillus casei, Lactobacillus reuteri, Bifidobacterium animalis lactis, and Lactobacillus rhamnosus in a mass ratio of 1.5:1.5:1.5:1.5. The concentration of Lactobacillus casei, Lactobacillus reuteri, Bifidobacterium animalis lactis, and Lactobacillus rhamnosus is 1.5 x 1011 cfu / ml.

[0053] The production method is referred to Example 1.

[0054] Example 3 The present example provides an oral liquid for removing damp-toxic and stagnated color spots. The probiotic fermented traditional Chinese medicine preparation is prepared by using probiotics to ferment a water extract of a traditional Chinese medicine composition as a fermentation substrate. The traditional Chinese medicine composition is prepared by mixing and extracting 20 parts of coicis, 20 parts of poria, 13.4 parts of winter melon seeds, 13.4 parts of chrysanthemum, 10 parts of prunella vulgaris, 10 parts of black beans, 10 parts of green beans, and 10 parts of licorice. The mass of the added mixed lactobacillus is 0.15% of the total mass of the above raw materials. The mixed lactobacillus includes lactobacillus casei, lactobacillus reuteri, animal bifidobacterium lactis subspecies, and lactobacillus rhamnosus in a mass ratio of 1:1:1:1. The concentrations of lactobacillus casei, lactobacillus reuteri, animal bifidobacterium lactis subspecies, and lactobacillus rhamnosus are 1.2×10 11 cfu / ml.

[0055] Comparative Example 1 The present comparative example provides an oral liquid for removing damp-toxic and stagnated color spots. The probiotic fermented traditional Chinese medicine preparation is prepared by using probiotics to ferment a water extract of a traditional Chinese medicine composition as a fermentation substrate. The traditional Chinese medicine composition and the water extraction process and centrifugal purification are the same as those in Example 3.

[0056] The centrifugal liquid of Comparative Example 1 and the fermentation liquid in step ③ of Example 3 are respectively taken, and the contents of glycerol trioleate, chlorogenic acid, chrysanthemum glycoside, 3,5-O-dicaffeoyl quinine, rosmarinic acid, glycyrrhizin, and glycyrrhizic acid are determined according to the method in Chinese Pharmacopoeia (2020 edition, part I). The test is repeated for three times, and the results show that only glycerol trioleate, chlorogenic acid, 3,5-O-dicaffeoyl quinine, glycyrrhizin, and glycyrrhizic acid are detected in the centrifugal liquid, while glycerol trioleate, chlorogenic acid, chrysanthemum glycoside, 3,5-O-dicaffeoyl quinine, rosmarinic acid, glycyrrhizin, and glycyrrhizic acid are detected in the fermentation liquid. It is indicated that the centrifugal liquid produces different effective components chrysanthemum glycoside and rosmarinic acid compared with the fermentation liquid. The specific results are shown in Table 1.

[0057] Table 1 Comparison of components in the centrifugal liquid of Comparative Example 1 and the fermentation liquid in step ③ of Example 3

[0058] Comparative Example 2 The present comparative example provides an oral liquid for removing damp-toxic and stagnated color spots. The probiotic fermented traditional Chinese medicine preparation is prepared by using probiotics to ferment a water extract of a traditional Chinese medicine composition as a fermentation substrate. The traditional Chinese medicine composition is the same as that in Example 3. The mass of the added mixed lactobacillus is 0.15% of the total mass of the above raw materials. The mixed lactobacillus includes lactobacillus casei and lactobacillus reuteri in a mass ratio of 1:1. The concentrations of lactobacillus casei and lactobacillus reuteri are 1.2×1011 cfu / ml.

[0059] The production method is referred to Example 3.

[0060] Comparative Example 3 This comparative example provides an oral liquid for removing damp-toxic and stagnated type of color spots, wherein the probiotic fermented traditional Chinese medicine preparation is prepared by using probiotics to ferment a water extract of a traditional Chinese medicine composition as a fermentation substrate; the traditional Chinese medicine composition is the same as that in Example 3; the added mixed lactobacillus has a mass of 0.15% of the total mass of the above raw materials. The mixed lactobacillus includes Lactobacillus casei, Lactobacillus reuteri and Bifidobacterium animalis ssp. lactis in a mass ratio of 1:1:1; the concentration of Lactobacillus casei, Lactobacillus reuteri and Bifidobacterium animalis ssp. lactis is 1.2 x 10 11 cfu / ml.

[0061] The production method is referred to Example 3.

[0062] Comparative Example 4 This comparative example provides an oral liquid for removing damp-toxic and stagnated type of color spots, wherein the probiotic fermented traditional Chinese medicine preparation is prepared by using probiotics to ferment a water extract of a traditional Chinese medicine composition as a fermentation substrate; the traditional Chinese medicine composition is the same as that in Example 3; the added mixed lactobacillus has a mass of 0.15% of the total mass of the above raw materials. The mixed lactobacillus includes Lactobacillus casei, Lactobacillus reuteri and Bifidobacterium animalis ssp. lactis in a mass ratio of 1:1:1; the concentration of Lactobacillus casei, Lactobacillus reuteri and Bifidobacterium animalis ssp. lactis is 1.2 x 10 11 cfu / ml.

[0063] The production method is referred to Example 3.

[0064] The inoculum of Example 3 and Comparative Examples 2-4 above is anaerobically cultured at 37±1°C (pH 6.5) for 24h, and samples are taken at 0h, 6h, 12h and 24h, respectively. The survival rate of each strain in the combination is evaluated by counting the viable bacteria (CFU / mL); the pH value of the fermentation broth, the yield of lactic acid / short-chain fatty acids (SCFAs) are detected; the degradation or conversion products of the target active ingredients (such as glycosides, flavonoids) are analyzed. The function is evaluated by taking the antioxidant capacity of the fermentation broth, the antibacterial activity, the bioavailability after in vitro simulated digestion as indexes. The differences between groups are compared (ANOVA), and the synergistic effect is verified by interaction analysis.

[0065] The experimental results show that the four-bacteria group shows obvious synergistic growth effect, L. rhamnosus promotes the survival of other strains, showing higher total viable count or longer stationary phase; the four-bacteria group has obvious synergistic effect of metabolic products, produces more abundant lactic acid / SCFAs, significantly reduces pH, can enhance the dissolution or stability of traditional Chinese medicine components, the conversion rate of specific components (such as glycyrrhizin) is higher, and more active metabolites (such as glycyrrhizin aglycone) are generated. The antioxidant / inhibitory activity of the four-bacteria fermentation broth is significantly better than that of other groups, indicating that the strains complement metabolic pathways and significantly improve bioavailability.

[0066] Comparative Example 5 The present comparative example provides an oral liquid for removing dampness-toxicity accumulation type color spots, the probiotic fermented traditional Chinese medicine preparation is prepared by using probiotics to ferment the water extract of a traditional Chinese medicine composition as a fermentation substrate; the traditional Chinese medicine composition is the same as that in Example 3; the added mixed lactobacillus has a mass of 0.15% of the total mass of the above raw materials. The mixed lactic acid bacteria are the same as those in Example 3.

[0067] The production method refers to Example 3, except that the inoculation ratio in step ③ is 1 / 100.

[0068] Comparative Example 6 The present comparative example provides an oral liquid for removing dampness-toxicity accumulation type color spots, the probiotic fermented traditional Chinese medicine preparation is prepared by using probiotics to ferment the water extract of a traditional Chinese medicine composition as a fermentation substrate; the traditional Chinese medicine composition is the same as that in Example 3; the added mixed lactobacillus has a mass of 0.15% of the total mass of the above raw materials. The mixed lactic acid bacteria are the same as those in Example 3.

[0069] The production method refers to Example 3, except that the inoculation ratio in step ③ is 2 / 100.

[0070] The fermentation broth in step ③ of the above Example 3 and Comparative Examples 5-6 is cultured anaerobically at 37±1°C for 24 hours, and sampled every 6 hours. The viable count, pH value, lactic acid content, polysaccharide consumption rate and content changes of target active ingredients (such as 3,5-O-dicaffeoyl quinic acid and glycyrrhizic acid) of the fermentation broth are detected. The results show that the peak viable count of the 1.5 / 100 group reaches the highest viable count (8.5×10 8 CFU / mL) at 36 hours, which is significantly higher than that of the 1.0 / 100 group (5.2×10 8 CFU / mL) and the 2.0 / 100 group (7.1×10 8CFU / mL), indicating that the 1.5 / 100 ratio is more conducive to the synergistic proliferation of the flora. The number of viable bacteria in the 2.0 / 100 group decreased later due to nutrient competition or metabolic product inhibition; lactic acid production: the lactic acid content in the 1.5 / 100 group (4.8 mg / mL) was higher than that in the other groups (1.0 / 100: 3.1 mg / mL; 2.0 / 100: 4.0 mg / mL), showing high glycolytic capacity; pH change: the pH of the 1.5 / 100 group stabilized at 4.2, which is consistent with the fermentation characteristics of probiotics; 3,5-O-dicaffeoylquinic acid bioconversion rate: the 1.5 / 100 group increased by 12.5% (control group 7.2%); fermentation broth stability: the 1.5 / 100 group had a 85% retention rate of viable bacteria after storage test (4°C, 14 days), which was better than the other groups (70%~78%).

[0071] The experimental results show that: 1.5 / 100 bacteria powder addition mass ratio balances the growth of the flora and substrate utilization, avoiding excessive proportion leading to resource competition; metabolic products (such as lactic acid) effectively promote the transformation of traditional Chinese medicine ingredients, while inhibiting miscellaneous bacteria. The 1.5 / 100 bacteria powder addition mass ratio is the optimal condition, with the highest viable cell density and metabolic activity; significantly improves the bioavailability of active ingredients; the stability of the fermentation broth is good.

[0072] Efficacy test experiment Examples 1-3 provide efficacy test experiments for the drink for removing liver stagnation and qi stagnation type color spots. The human test was conducted according to the cosmetic and color spot removal function test method of the “Health Food Function Test and Evaluation Technical Guidelines (2023 Edition)” and “Human Open Use Test Color Spot Removal and Whitening Efficacy Evaluation Method”. The specific experimental steps are as follows: 1. Inclusion criteria for subjects Volunteer subjects who meet the following conditions after examination: (1) There are light brown to dark brown, well-defined patches on the face, usually symmetrical distribution, no inflammatory manifestations and scales; (2) The ITA of the pigmented skin and the surrounding adjacent skin o Difference is greater than 10; (3) No obvious subjective symptoms; (4) No obvious endocrine diseases, and exclude pigmentation caused by other diseases.

[0073] 2. Exclusion criteria for subjects (1) Age below 20 years old and above 60 years old, pregnant and lactating women, and people with allergic constitution; (2) Combined with severe diseases of cardiovascular, cerebrovascular, liver, kidney and hematopoietic system, and endocrine diseases, and patients with mental illness; (3) Alcoholics or smokers; (4) Taking products related to the function of the test within a short period of time, which affects the judgment of the results; (5) The test sample is not taken as required, and the efficacy or safety judgment is affected by incomplete data.

[0074] 3. Test design and grouping requirements A normal dose group and a positive control group are designed, and an ineffective placebo (pure water) is used as a negative control. According to the color and area of facial spots of the subjects, the subjects are randomly divided into a normal dose group, a positive control group and a negative control group. The main factors affecting the results such as outdoor activities, gender, age, etc. are considered as much as possible, and the balance test is performed to ensure the comparability between groups.

[0075] The test substance is a medicinal and edible product for removing liver stagnation and qi stagnation type spots, which is prepared according to the formula and process of the present application. 120 female subjects (40 subjects on average in each group, 2 subjects in the negative control group) meeting the above conditions are selected according to the voluntary principle. The normal dose group takes 1 bag of the fermented medicinal drink each time, 2 times a day, with a dosage of 60 mL / day, for 30 consecutive days; the positive control group takes 1 bag of powder prepared according to the invention (patent number: CN120022332A), which is brewed in 50 mL of warm water and then drunk, 2 times a day, for 30 consecutive days; the negative control group takes the same amount of ineffective placebo as the normal dose group. The subjects stop using the spot-removing products, drugs and health foods for beauty and spot removal during the test period.

[0076] 4. Detection of facial spots 4.1 Change in area size The length and width of the same facial spot before and after the test are measured with a ruler, and the area (cm 2 ) is calculated.

[0077] 4.2 Change in color depth According to the brown (Y+M+Bk, i.e. yellow+magenta+black) color card in the Practical Standard Color Card (first edition) as the judgment standard for the color depth of facial spots: I degree (15, 20, 5), score ≤1.0; II degree (30, 40, 10), 1.0<score ≤2.0; III degree (40, 60, 15), score >2.0. The values in the brackets represent the color percentage concentration values of yellow, magenta and black respectively.

[0078] SPSS (Social Science Statistics Software Package) is used to statistically analyze the changes in the area and color of facial spots of the subjects. The test data are shown in Tables 2-7.

[0079] Table 2: Test results of spot area in the normal dose group

[0080] Table 3: Spot area detection results of the positive control group

[0081] Table 4: Spot area detection results of the negative control group

[0082] Table 5: Spot color detection results of the normal dose group

[0083] Table 6: Spot color detection results of the positive control group

[0084] Table 7: Spot color detection results of the negative control group

[0085] Note: Y represents significant or effective, and N represents ineffective.

[0086] 5.1 Analysis of spot-removing efficacy of the oral liquid for removing damp-toxic and stagnated spots 5.1.2 The results of changes in the area of facial spots are shown in Table 8: Table 8: Changes in the area of facial spots of the test group and the control group

[0087] Note: Intra-group comparison p<0.01, containing effective and significant number of cases.

[0088] 5.1.3 The changes in the color depth of facial spots are shown in Table 9: Table 9: Changes in the color depth of facial spots of the test group and the control group

[0089] Note: Intra-group comparison p<0.01, containing effective and significant number of cases.

[0090] 5.1.4 The results of efficacy determination before and after the test (cases, %) are shown in Table 10: Table 10: Efficacy determination results of the test group and the control group before and after the test

[0091] 5.1.5 Test conclusion The average area of facial spots of 40 subjects with facial spots was reduced by 45.15 cm 2(P<0.01), the color becomes light, the color card score average reduces 0.70 (P<0.01), wherein effective example is 33, total efficiency is 82.50%, and the effective rate of the positive control group is only 21.50%. It shows that the medicine-food homologous freckle-removing oral liquid has a significant effect of removing dampness-toxicity accumulation type color spots.

[0092] The above embodiments are the preferred embodiments of the present application, but the embodiments of the present application are not limited to the above embodiments, and any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application should be equivalent replacement methods, and all are included in the protection scope of the present application.

Claims

1. A probiotic fermented traditional Chinese medicine preparation, characterized in that, The probiotic fermented traditional Chinese medicine preparation is prepared by using probiotics to ferment a water extract of a traditional Chinese medicine composition as a fermentation substrate; The traditional Chinese medicine composition comprises coix seed, poria cocos, winter melon seed, chrysanthemum, prunella vulgaris, black bean, green bean, and licorice; The probiotics comprise Lactobacillus casei, Lactobacillus reuteri, Bifidobacterium animalis ssp. lactis, and Lactobacillus rhamnosus.

2. The probiotic fermented traditional Chinese medicine preparation according to claim 1, characterized in that, The traditional Chinese medicine composition comprises 10-30 parts of coix seed, 10-30 parts of poria cocos, 5-20 parts of winter melon seed, 5-20 parts of chrysanthemum, 5-15 parts of prunella vulgaris, 5-15 parts of black bean, 5-15 parts of green bean, and 5-15 parts of licorice by mass; The probiotics comprise 0.5-1.5 parts of Lactobacillus casei, 0.5-1.5 parts of Lactobacillus reuteri, 0.5-1.5 parts of Bifidobacterium animalis ssp. lactis, and 0.5-1.5 parts of Lactobacillus rhamnosus by mass.

3. The probiotic fermented traditional Chinese medicine preparation according to claim 1, characterized in that, The mass ratio of the probiotics to the water extract of the traditional Chinese medicine composition is (1-2):100; The mass ratio of the probiotics to the traditional Chinese medicine composition is (0.01-0.3):

100.

4. The probiotic fermented traditional Chinese medicine preparation according to claim 1, characterized in that, The viable cell count of the Lactobacillus casei, Lactobacillus rhamnosus, Bifidobacterium animalis ssp. lactis and Lactobacillus rhamnosus is not less than 1 x 10 10 CFU / mL.

5. The method for preparing the probiotic fermented traditional Chinese medicine preparation according to any one of claims 1-4, characterized in that, The method comprises the following steps: The coix seed, poria cocos, winter melon seed, chrysanthemum, prunella vulgaris, black bean, green bean, and licorice are mixed and water-extracted to obtain a water extract; the water extract and the probiotics are placed in a fermentation tank to be fermented to prepare the probiotic fermented traditional Chinese medicine preparation.

6. The method of claim 5, wherein the preparation of the probiotic fermented traditional Chinese medicine preparation is characterized by, The water extraction is performed at a temperature of 90-100℃.

7. The method of claim 6, wherein the preparation of the probiotic fermented traditional Chinese medicine preparation is characterized by, The other process parameters of the water extraction are as follows: the solid-liquid weight ratio is 1:(6-10); the extraction times are 1-2 times; and the single extraction time is 1-2 h.

8. The method of claim 5, wherein the preparation of the probiotic fermented traditional Chinese medicine preparation is characterized by, The method further comprises the following step: after the water extraction, the water extract is centrifuged to obtain a centrifugate.

9. The method of claim 5, wherein the preparation of the probiotic fermented traditional Chinese medicine preparation is characterized by, The fermentation is performed at a temperature of 36-38℃; The fermentation is performed for a time of 16-30 h.

10. Use of the probiotic fermented traditional Chinese medicine preparation according to any one of claims 1-4 in the preparation of a medicament for treating dampness-heat accumulation or a symptom caused thereby.

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