Ginseng and rhizoma polygonati fermentation composition and nourishing application

Abstract

The invention relates to the field of medicinal and edible traditional Chinese medicine fermentation, in particular to a ginseng and rhizoma polygonati fermentation composition and nourishing application. The invention solves the problem of poor efficacy of a medicinal and edible traditional Chinese medicine formula. According to the medicinal and edible traditional Chinese medicine fermented composition provided by the invention, the efficacy of the composition is improved by about 2 times compared with that of the unfermented composition; the invention also provides a preparation method of the fermentation composition. The preparation method comprises an optimal extraction process, an optimal strain, an optimal strain combination and an optimal fermentation process. In addition, the invention also discloses an optimal traditional Chinese medicine formula with homology of medicine and food through formula screening research.

Description

Technical Field

[0001] The invention relates to the field of traditional Chinese medicine fermentation, and in particular to a ginseng and polygonatum fermentation composition and a tonic application thereof. Background Art

[0002] Insufficient Qi and blood can lead to problems and diseases such as low immunity, fatigue and chronic fatigue syndrome. The nourishing and regulating treatment of Chinese medicine with the same origin of medicine and food can have a significant effect. Chronic fatigue syndrome (CFS) can occur in people of any age and race, seriously affecting the work and life of patients. Its incidence rate is increasing year by year and has become a key disease of concern in the medical community. Studies have shown that the incidence of chronic fatigue syndrome in the United States is 0.1% to 0.2%, and the incidence rate in Japan is 1.5%. Experts predict that it will become one of the major problems affecting human health in the 21st century. Therefore, health intervention is necessary and effective prevention and treatment measures are urgently needed. However, there is still a lack of specific therapeutic drugs. Moreover, the current products for nourishing, enhancing immunity, resisting fatigue, and preventing and treating chronic fatigue syndrome need to be taken for a long time, with great side effects, resulting in "disease A is not cured, and disease B is added". Chinese medicine with the same origin of medicine and food is very suitable for nourishing, enhancing immunity, resisting fatigue, and preventing and treating chronic fatigue syndrome due to its high safety. However, the main problem of Chinese medicine with the same origin of medicine and food is that the formula is unscientific or the efficacy is low. Therefore, it is urgently needed to invent the best Chinese medicine formula for food and medicine with the functions of nourishing, enhancing immunity, resisting fatigue, preventing and treating chronic fatigue syndrome, and improving the efficacy. Summary of the invention

[0003] In view of this, the present invention provides a ginseng and polygonatum fermentation composition and a tonic application. The present invention solves the problem of poor efficacy of Chinese medicine prescriptions that are both medicinal and edible. On the one hand, the present invention improves the efficacy of Chinese medicines that are both medicinal and edible in chronic fatigue syndrome, tonic, enhancing immunity, anti-fatigue, etc. by fermentation technology, and improves it by about 2 times. On the other hand, through prescription screening research, the best Chinese medicine prescription that is both medicinal and edible is invented.

[0004] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0005] In a first aspect, the present invention provides a Chinese medicinal fermentation composition that is both medicinal and edible, comprising ginseng, angelica, jujube, ginger, polygonatum, kudzu root, tuckahoe, tangerine peel, cinnamon and liquorice;

[0006] The bacteria used in the fermentation include probiotics.

[0007] In some specific embodiments of the present invention, the Chinese medicinal fermentation composition comprising, by weight:

[0008]

[0009] Preferably, the edible and medicinal Chinese medicine fermentation composition comprises, by weight:

[0010] 50-130 parts of ginseng, 50-130 parts of angelica, 50-130 parts of jujube, 50-130 parts of ginger, 30-90 parts of polygonatum, 30-90 parts of kudzu root, 30-90 parts of tuckahoe, 30-90 parts of tangerine peel, 20-40 parts of cinnamon and 20-40 parts of liquorice; or

[0011] More preferably, the edible and medicinal Chinese medicine fermentation composition comprises, by weight:

[0012] 60-110 parts of ginseng, 60-110 parts of angelica, 60-110 parts of jujube, 60-110 parts of ginger, 40-80 parts of polygonatum, 40-80 parts of kudzu root, 40-80 parts of tuckahoe, 40-80 parts of tangerine peel, 20-30 parts of cinnamon and 20-30 parts of liquorice; or

[0013] More preferably, the edible and medicinal Chinese medicine fermentation composition comprises, by weight:

[0014] 60-90 parts of ginseng, 60-90 parts of angelica, 60-90 parts of jujube, 60-90 parts of ginger, 40-60 parts of polygonatum, 40-60 parts of kudzu root, 40-60 parts of poria, 40-60 parts of tangerine peel, 20-30 parts of cinnamon and 20-30 parts of liquorice.

[0015] In some specific embodiments of the present invention, the Chinese medicinal fermentation composition comprising, by weight:

[0016] 75 parts of ginseng, 75 parts of angelica, 75 parts of jujube, 75 parts of ginger, 50 parts of polygonatum, 50 parts of kudzu root, 50 parts of tuckahoe, 50 parts of tangerine peel, 25 parts of cinnamon and 25 parts of liquorice; or

[0017] 130 parts of ginseng, 50 parts of angelica, 130 parts of jujube, 50 parts of ginger, 90 parts of polygonatum, 40 parts of kudzu root, 40 parts of tuckahoe, 60 parts of tangerine peel, 30 parts of cinnamon and 20 parts of liquorice; or

[0018] 90 parts of ginseng, 90 parts of angelica, 60 parts of jujube, 60 parts of ginger, 40 parts of polygonatum, 60 parts of kudzu root, 60 parts of tuckahoe, 40 parts of tangerine peel, 20 parts of cinnamon and 30 parts of liquorice; or

[0019] 75 parts of ginseng, 75 parts of angelica, 75 parts of jujube, 30 parts of ginger, 50 parts of polygonatum, 50 parts of kudzu root, 50 parts of tuckahoe, 50 parts of tangerine peel, 25 parts of cinnamon and 25 parts of liquorice; or

[0020] 75 parts of the ginseng, 75 parts of the angelica, 75 parts of the jujube, 140 parts of the ginger, 50 parts of the polygonatum, 50 parts of the kudzu root, 50 parts of the tuckahoe, 50 parts of the tangerine peel, 25 parts of the cinnamon and 25 parts of the liquorice.

[0021] In some specific embodiments of the present invention, the ginseng in the edible and medicinal Chinese medicine fermentation composition includes ginseng extract and / or ginseng concentrate; or

[0022] More preferably, the ginseng extract or the ginseng concentrate is obtained by extracting ginseng and water in a ratio of 3 to 3.5:1 (W / W); the water includes drinking water; or

[0023] More preferably, the content of total ginsenosides in the ginseng extract or the ginseng concentrate is ≥3%.

[0024] In some specific embodiments of the present invention, the probiotics include one or more of the genera Lactobacillus, Lactobacillus, Lactobacillus and / or Lactobacillus mucilaginosus; or

[0025] More preferably, the genus Lactobacillus includes one or more of Lactobacillus casei, Lactobacillus paracasei and / or Lactobacillus rhamnosus; the genus Lactobacillus includes Lactobacillus plantarum; the genus Lactobacillus includes one or more of Lactobacillus delbrueckii subspecies bulgaricus and / or Lactobacillus helveticus; the genus Lactobacillus mucosus includes Lactobacillus fermentum; or

[0026] More preferably, the probiotics include the Lactobacillus genus and the Lactobacillus genus; or

[0027] More preferably, the probiotics include Lactobacillus plantarum and Lactobacillus rhamnosus; or

[0028] More preferably, the inoculation amount of the probiotics includes 2-10% (V / V); preferably, the inoculation amount of the probiotics includes 2%, 3%, 4%, 5% or 6% (V / V); more preferably, the inoculation amount of the probiotics includes 4% (V / V);

[0029] Or the inoculation ratio of the Lactobacillus genus and the Lactobacillus genus in the probiotics includes (1-3): (1-3); or

[0030] Preferably, the inoculation ratio of the plant lactobacillus and the rhamnosus lactobacillus in the probiotics is 1:1; the OD of the plant lactobacillus in the probiotics is600 Including 3.0 to 5.0, the OD of the rhamnosus Lactobacillus 600 3.5 to 5.5 inclusive; or

[0031] The pH of the probiotic inoculation comprises 6.5; or

[0032] The fermentation time of the probiotic fermentation includes 16 to 48 hours; or

[0033] Preferably, the fermentation time of the probiotic fermentation includes 20 hours, 24 hours, 26 hours, 28 hours or 32 hours; preferably, the fermentation time of the probiotic fermentation includes 24 to 32 hours.

[0034] In a second aspect, the present invention also provides a method for preparing the edible and medicinal Chinese medicine fermentation composition, wherein the raw materials are weighed according to the ratio, and the edible and medicinal Chinese medicine fermentation composition is obtained through extraction, concentration, fermentation, blending and filling;

[0035] The extraction includes but is not limited to 1 extraction or 2 extractions; or

[0036] Preferably, the extraction includes one extraction; preferably, the amount of water added in the one extraction includes 8 to 12 times the amount of the raw material; more preferably, the amount of water added in the one extraction includes 10 times the amount of the raw material; preferably, the time for one extraction is 1 to 3 hours; more preferably, the time for one extraction is 1.5 hours; or

[0037] Preferably, the extraction comprises 2 extractions;

[0038] The amount of water added in the two extractions includes 14 to 22 times of the raw material, the amount of water added in the first of the two extractions includes 8 to 12 times of the raw material, and the amount of water added in the second of the two extractions includes 6 to 10 times of the raw material;

[0039] Preferably, the amount of water added in the first extraction of the two extractions includes 10 or 12 times the amount of the raw material, and the amount of water added in the second extraction of the two extractions includes 8 times the amount of the raw material;

[0040] More preferably, the amount of water added in the first extraction of the two extractions includes 10 times the amount of the raw material;

[0041] The time for the two extractions includes 1 to 3 hours, the time for the first extraction of the two extractions includes 1 to 2 hours, and the time for the second extraction of the two extractions includes 1 to 2 hours;

[0042] Preferably, the time for the first extraction of the two extractions includes 1 hour, 1.5 hours or 2 hours, and the time for the second extraction of the two extractions includes 1 hour;

[0043] The concentration includes but is not limited to double-effect concentration; the multiple of concentration includes but is not limited to 3 times;

[0044] The fermentation temperature includes but is not limited to 37° C., the rotation speed includes but is not limited to 50 rpm, the tank pressure includes 0.04-0.06 MPa; and the pH at the end of the fermentation includes 3.3-3.8.

[0045] In a third aspect, the present invention also provides a Chinese medicinal fermentation composition that is both medicinal and edible, obtained by the preparation method.

[0046] In a fourth aspect, the present invention also provides a compound fermented product of a Chinese medicine composition that is both a medicine and food, comprising the Chinese medicine fermented composition that is both a medicine and food;

[0047] Preferably, the compound fermented product of the Chinese medicine composition with the same origin as food and medicine further comprises one or more of oligofructose, γ-aminobutyric acid and / or taurine; the oligofructose comprises oligofructose powder and / or oligofructose liquid; or

[0048] Preferably, the compound fermented product of the Chinese medicine composition with both medicinal and edible properties further comprises one or more of erythritol, xylitol and / or black wolfberry juice powder.

[0049] In some specific embodiments of the present invention, the compound fermented product of the Chinese medicine composition of food and medicine also includes, by weight:

[0050] 100-700 parts of the oligofructose, 5-20 parts of the γ-aminobutyric acid and / or 2-10 parts of the taurine;

[0051] Preferably, the compound fermented product of the Chinese medicine composition with the same origin as food and medicine further comprises, by weight:

[0052] 200-500 parts of the oligofructose, 5-10 parts of the γ-aminobutyric acid and / or 2-5 parts of the taurine;

[0053] More preferably, the compound fermented product of the Chinese medicine composition with the same origin as food and medicine further comprises, by weight:

[0054] 100-300 parts of the oligofructose, 5-10 parts of the γ-aminobutyric acid and / or 2-5 parts of the taurine;

[0055] More preferably, the compound fermented product of the Chinese medicine composition with the same origin as food and medicine further comprises, by weight:

[0056] 350 parts of the oligofructose, 10 parts of the γ-aminobutyric acid and / or 5 parts of the taurine; or

[0057] 250 parts of the oligofructose, 10 parts of the γ-aminobutyric acid and / or 5 parts of the taurine; or

[0058] 200 parts of the oligofructose, 5 parts of the gamma-aminobutyric acid and / or 2 parts of the taurine; or

[0059] 300 parts of the oligofructose, 20 parts of the gamma-aminobutyric acid and / or 5 parts of the taurine.

[0060] Preferably, the compound fermented product of the Chinese medicine composition with the same origin as food and medicine further comprises, by weight:

[0061] 100-400 parts of erythritol, 50-200 parts of xylitol and / or 50-200 parts of black wolfberry juice powder;

[0062] Preferably, the compound fermented product of the Chinese medicine composition with the same origin as food and medicine further comprises, by weight:

[0063] 150-300 parts of erythritol, 80-150 parts of xylitol and / or 80-150 parts of black wolfberry juice powder;

[0064] More preferably, the compound fermented product of the Chinese medicine composition with the same origin as food and medicine further comprises, by weight:

[0065] 200 parts of the erythritol, 100 parts of the xylitol and / or 100 parts of the black wolfberry juice powder; or

[0066] 100 parts of the erythritol, 150 parts of the xylitol and / or 150 parts of the black wolfberry juice powder; or

[0067] 300 parts of the erythritol, 80 parts of the xylitol and / or 80 parts of the black wolfberry juice powder.

[0068] Preferably, the compound fermented product of the Chinese medicine composition with the same origin as food includes medicine or food; more preferably, the food includes one or more of health food, food for special medical purposes, functional food or pet food.

[0069] In a fifth aspect, the present invention further provides the use of any of the following in the preparation of a tonic, anti-fatigue, immunity-enhancing and / or anti-oxidative product:

[0070] (I), the Chinese medicinal fermentation composition having both medicinal and edible properties;

[0071] (II), the Chinese medicinal fermentation composition having both medicinal and edible properties; or

[0072] (III) The compound fermented product of the Chinese medicine composition having both medicinal and edible properties.

[0073] In some specific embodiments of the present invention, the anti-fatigue comprises anti-chronic fatigue syndrome;

[0074] The anti-oxidation includes increasing the total antioxidant activity and / or increasing the cellular reduced glutathione content;

[0075] The improving of the total antioxidant activity comprises improving one or more of the following: the activity of superoxide dismutase in cells, the scavenging rate of DPPH free radicals and / or the scavenging rate of hydroxyl free radicals;

[0076] The immunity enhancement includes but is not limited to increasing the cell phagocytic index;

[0077] Preferably, the product includes medicine or food; more preferably, the food includes one or more of health food, food for special medical purposes, functional food or pet food.

[0078] In a sixth aspect, the present invention also provides a product, comprising any of the following items and acceptable excipients;

[0079] (I), the Chinese medicinal fermentation composition having both medicinal and edible properties;

[0080] (II), the Chinese medicinal fermentation composition having both medicinal and edible properties; or

[0081] (III) The compound fermented product of the Chinese medicine composition having both medicinal and edible properties.

[0082] In a seventh aspect, the present invention further provides a method for nourishing, resisting fatigue, enhancing immunity and / or resisting oxidation, the method comprising administering or using any of the following:

[0083] (I), the aforementioned Chinese medicinal fermentation composition having both medicinal and edible properties; and / or

[0084] (II), the Chinese medicinal fermentation composition having both medicinal and edible properties; and / or

[0085] (III), the compound fermented product of the Chinese medicine composition with the same function as food and medicine; and / or

[0086] (IV) the product described.

[0087] The present invention solves the problem of poor efficacy of Chinese medicine formulas that are both medicinal and edible. The present invention provides a Chinese medicine fermentation composition that is both medicinal and edible, and the efficacy of the composition is improved by about 2 times compared with that before fermentation; the present invention also provides a preparation method of the fermentation composition, including an optimal extraction process, an optimal strain, an optimal strain combination and an optimal fermentation process; in addition, the present invention also invents an optimal Chinese medicine formula that is both medicinal and edible through formula screening research.

[0088] The beneficial effects of the present invention include:

[0089] 1. Fermentation strains: Not all strains can be fermented. Lactobacillus delbrueckii subspecies bulgaricus and fermentation Lactobacillus mucilaginosus are not suitable for growth (see strain study in the embodiment); the present invention screens out a strain combination of Lactobacillus plantarum and Lactobacillus rhamnosus for fermentation of traditional Chinese medicine prescriptions;

[0090] 2. Traditional Chinese medicine fermentation formula: Compared with before fermentation, the antioxidant, immune-enhancing and anti-fatigue functions of the traditional Chinese medicine formula after fermentation are significantly improved (see Example);

[0091] 3. Traditional Chinese medicine fermentation formula: Not all bacterial strain combinations improve the effect. The coordination effect between the bacterial strain combination of Lactobacillus plantarum, Lactobacillus rhamnosus and Lactobacillus helveticus, and the bacterial strain combination of Lactobacillus plantarum and Lactobacillus helveticus is poor (see Examples).

[0092] 4. Traditional Chinese medicine fermentation formula + blending efficacy formula: better than Bazhenye, a drug for nourishing, replenishing qi and blood, and treating chronic fatigue syndrome, and the anti-fatigue health food that has been launched on the market; the human trials of nourishing, relieving fatigue, and 240 clinical trials of preventing and treating chronic fatigue syndrome have significant results;

[0093] 5. Traditional Chinese medicine fermentation formula + preparation of efficacy formula: The dosage of some main medicinal materials of the product of the present invention is increased by 2, 3, or 4 times, and fermentation is carried out according to the process of the present invention. The antioxidant function is not significantly improved in proportion, indicating that the formula of the present invention is the best formula, which saves the dosage of medicinal materials, finds the best ratio, and achieves unexpected results (see the examples). The existing formula is superior to the formula ratio outside the scope of the present invention. DETAILED DESCRIPTION

[0094] The present invention discloses a ginseng and polygonatum fermentation composition and a tonic application. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It should be particularly noted that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The method and application of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0095] Traditional Chinese medicines that are both edible and medicinal are very suitable for applications in tonic, chronic fatigue syndrome, enhancing immunity, anti-fatigue, anti-oxidation, etc. due to their high safety. However, the main problem of traditional Chinese medicines that are both edible and medicinal is their low efficacy. The present invention relates to a fermented traditional Chinese medicine composition that is both edible and medicinal, as well as a preparation method and tonic application of the fermented composition. The tonic application includes enhancing immunity, anti-fatigue, and preventing and treating chronic fatigue syndrome.

[0096] The Chinese medicine fermentation composition of the present invention mainly comprises ginseng, angelica, jujube, ginger, polygonatum, kudzu root, poria, tangerine peel, cinnamon and liquorice. Formula idea: ginseng, polygonatum and poria are the main drugs in the formula, which can greatly replenish the vital energy, replenish the kidney essence, strengthen the spleen and replenish qi, and strengthen the acquired foundation, so as to make the vital energy sufficient; the ministers are angelica and jujube, which can replenish blood and promote blood circulation, enhance the effects of the main drugs in nourishing, enhancing immunity, resisting fatigue and preventing and treating chronic fatigue syndrome; tangerine peel, cinnamon and ginger are used as adjuvants, tangerine peel can regulate qi and relieve stagnation, and assist the transportation and transformation function of the spleen, cinnamon can warm and dredge the meridians, guide fire back to the origin, ginger can relieve the exterior and dispel cold, and the three drugs are used together to make the main drugs and minister drugs nourishing but not greasy, so as to help the biochemical transformation of qi and blood; kudzu root and liquorice are the guiding drugs, kudzu root can promote the meridians and activate the collaterals, and encourage the spleen and stomach to rise, and liquorice can nourish the spleen and replenish qi and harmonize the drugs. The compound preparation is made by fermenting the Chinese medicinal materials with the same medicinal and edible properties, and the fermentation bacteria used include probiotics.

[0097] The composition comprises ginseng, angelica, jujube, ginger, polygonatum, kudzu root, tuckahoe, tangerine peel, cinnamon and liquorice. 30-140 parts by weight of ginseng, 30-140 parts by weight of angelica, 30-140 parts by weight of jujube, 30-140 parts by weight of ginger, 20-90 parts by weight of polygonatum, 20-90 parts by weight of kudzu root, 20-90 parts by weight of tuckahoe, 20-90 parts by weight of tangerine peel, 10-45 parts by weight of cinnamon and 10-45 parts by weight of liquorice.

[0098] The above composition includes 50-130 parts by weight of ginseng, 50-130 parts by weight of angelica, 50-130 parts by weight of jujube, 50-130 parts by weight of ginger, 30-90 parts by weight of polygonatum, 30-90 parts by weight of kudzu root, 30-90 parts by weight of poria, 30-90 parts by weight of tangerine peel, 20-40 parts by weight of cinnamon, and 20-40 parts by weight of liquorice.

[0099] The above composition includes 60-110 parts by weight of ginseng, 60-110 parts by weight of angelica, 60-110 parts by weight of jujube, 60-110 parts by weight of ginger, 40-80 parts by weight of polygonatum, 40-80 parts by weight of kudzu root, 40-80 parts by weight of poria, 40-80 parts by weight of tangerine peel, 20-30 parts by weight of cinnamon, and 20-30 parts by weight of licorice.

[0100] The above composition includes 60-90 parts by weight of ginseng, 60-90 parts by weight of angelica, 60-90 parts by weight of jujube, 60-90 parts by weight of ginger, 40-60 parts by weight of polygonatum, 40-60 parts by weight of kudzu root, 40-60 parts by weight of poria, 40-60 parts by weight of tangerine peel, 20-30 parts by weight of cinnamon, and 20-30 parts by weight of liquorice.

[0101] The composition comprises 75 parts by weight of ginseng, 75 parts by weight of angelica, 75 parts by weight of jujube, 75 parts by weight of ginger, 50 parts by weight of polygonatum, 50 parts by weight of kudzu root, 50 parts by weight of tuckahoe, 50 parts by weight of tangerine peel, 25 parts by weight of cinnamon and 25 parts by weight of liquorice.

[0102] The composition comprises 130 parts by weight of ginseng, 50 parts by weight of angelica, 130 parts by weight of jujube, 50 parts by weight of ginger, 90 parts by weight of polygonatum, 40 parts by weight of kudzu root, 40 parts by weight of poria, 60 parts by weight of tangerine peel, 30 parts by weight of cinnamon and 20 parts by weight of liquorice.

[0103] The composition comprises 90 parts by weight of ginseng, 90 parts by weight of angelica, 60 parts by weight of jujube, 60 parts by weight of ginger, 40 parts by weight of polygonatum, 60 parts by weight of kudzu root, 60 parts by weight of poria, 40 parts by weight of tangerine peel, 20 parts by weight of cinnamon and 30 parts by weight of liquorice.

[0104] The ginseng further includes ginseng extract (ginseng concentrate).

[0105] The ginseng extract (ginseng concentrate) is obtained by extracting ginseng and drinking water in a ratio of 3 to 3.5:1 (W / W), and the total ginsenosides are ≥3%.

[0106] The probiotics of the present invention include Lactobacillus, Lactobacillus, Lactobacillus and Lactobacillus mucilaginosus.

[0107] The Lactobacillus genus described in the present invention includes Lactobacillus casei, Lactobacillus paracasei and Lactobacillus rhamnosus.

[0108] The Lactobacillus genus of the present invention includes Lactobacillus plantarum.

[0109] The Lactobacillus genus described in the present invention includes Lactobacillus delbrueckii subspecies bulgaricus and Lactobacillus helveticus.

[0110] The genus Lactobacillus mucilaginosus of the present invention includes fermentative Lactobacillus mucilaginosus.

[0111] The inoculation amount of the probiotics of the present invention is 2-10%, more preferably 2%, 3%, 4%, 5% or 6%.

[0112] The pH value of the probiotics during inoculation of the present invention is 6.5.

[0113] The fermentation time of the probiotic fermentation of the present invention is 16 to 42 hours, and more preferably 20 hours, 24 hours, 26 hours, 28 hours and 32 hours.

[0114] The above-mentioned Chinese medicinal fermentation composition which is both edible and medicinal may also include one or more of oligofructose, γ-aminobutyric acid or taurine, and the oligofructose includes oligofructose powder or oligofructose liquid. The invention further comprises 100-700 parts by weight of oligofructose, 5-20 parts by weight of γ-aminobutyric acid or 2-10 parts by weight of taurine, further preferably 200-500 parts by weight of oligofructose, 5-10 parts by weight of γ-aminobutyric acid or 2-5 parts by weight of taurine, further preferably 100-300 parts by weight of oligofructose, 5-10 parts by weight of γ-aminobutyric acid or 2-5 parts by weight of taurine, further preferably 250 parts by weight of oligofructose, 10 parts by weight of γ-aminobutyric acid or 5 parts by weight of taurine, further preferably 350 parts by weight of oligofructose, 10 parts by weight of γ-aminobutyric acid or 5 parts by weight of taurine, further preferably 200 parts by weight of oligofructose, 5 parts by weight of γ-aminobutyric acid or 2 parts by weight of taurine, further preferably 300 parts by weight of oligofructose, 20 parts by weight of γ-aminobutyric acid or 5 parts by weight of taurine.

[0115] The above-mentioned Chinese medicinal fermentation composition with both medicinal and edible properties also includes one or more of erythritol, xylitol, and black wolfberry juice powder, further including 100-400 weight parts of erythritol, 50-200 weight parts of xylitol, and 50-200 weight parts of black wolfberry juice powder, further including 150-300 weight parts of erythritol, 80-150 weight parts of xylitol, and 80-150 weight parts of black wolfberry juice powder, further including 200 weight parts of erythritol, 100 weight parts of xylitol, and 100 weight parts of black wolfberry juice powder, further including 100 weight parts of erythritol, 150 weight parts of xylitol, and 150 weight parts of black wolfberry juice powder, further including 300 weight parts of erythritol, 80 weight parts of xylitol, and 80 weight parts of black wolfberry juice powder.

[0116] The preparation process of the edible and medicinal Chinese medicine fermentation composition of the present invention comprises extraction, concentration, fermentation, preparation and filling.

[0117] The extraction process preferably includes two extractions; the two extractions preferably include 14 to 22 times of water, more preferably 8 to 12 times of water for the first extraction and 6 to 10 times of water for the second extraction, more preferably 10 or 12 times of water for the first extraction and 8 times of water for the second extraction.

[0118] The extraction process, the extraction process, two extractions, preferably the extraction time is 1 to 3 hours, more preferably the first extraction is 1 to 2 hours, the second extraction is 1 to 2 hours, more preferably the first extraction is 1, 1.5 hours or 2 hours, the second extraction is 1 hour.

[0119] The product of the present invention is preferably prepared according to the weight ratio of the present invention and processed according to the preparation process of the present invention into 50 mL per bottle, and 1 to 2 bottles per day are recommended.

[0120] The raw materials and reagents used in the ginseng and polygonatum fermentation composition and the nourishing application provided by the invention can all be purchased from the market.

[0121] The present invention will be further described below in conjunction with embodiments:

[0122] Example 1 Extraction process study

[0123] 1. Formulation

[0124] The formula of the Chinese medicine composition is shown in Table 1.

[0125] Table 1 Chinese medicine composition formula

[0126]

[0127]

[0128] 2. Extraction process research

[0129] (1) The extraction solvent is drinking water

[0130] (2) Detection indicators: total flavonoids (detection method: 2020 Method 1 for the determination of total flavonoids in health foods) and total crude polysaccharides (detection method: 2015- Determination of crude polysaccharides in exported plant-based foods - phenol-sulfuric acid method).

[0131] 2.1 Screening of solid-liquid ratio for one extraction

[0132] 2.1.1、Material-liquid ratio group setting

[0133] The solid-liquid ratios (W / W or W / V) are 1:8, 1:9, 1:10, 1:11, and 1:12, respectively.

[0134] 2.1.2 Extraction method

[0135] Soak at 45℃ for 1h, boil and extract for 2h, then pass through a 100-mesh sieve to obtain the extract.

[0136] 2.1.3 Results

[0137] (1) The results of crude polysaccharides and total flavonoids are shown in Table 2.

[0138] Table 2 Detection results of crude polysaccharides and total flavonoids

[0139] Serial number Material-liquid ratio Total crude polysaccharides, mg Total flavonoids, mg 1 1:8 1089.6 44.8 2 1:9 1056.6 54.9 3 1:10 1215 63 4 1:11 1083.5 51.7 5 1:12 1084.8 68.4

[0140] (2) Results: Based on the comprehensive analysis of the contents of crude polysaccharides and total flavonoids, the optimal solid-liquid ratio for a single extraction is 1:10.

[0141] 2.2. Extraction time screening for one extraction

[0142] 2.2.1. Extraction time group settings

[0143] The extraction times are 1h, 1.5h, 2.0h, 2.5h and 3.0h respectively.

[0144] 2.2.2 Extraction method

[0145] Add water at a solid-liquid ratio of 1:10, raise the temperature to 45°C and soak for 1 hour, boil and extract for 1 hour, 1.5 hours, 2.0 hours, 2.5 hours, and 3.0 hours respectively, and then pass through a 100-mesh sieve to obtain the extract.

[0146] 2.2.3 Results

[0147] (1) The results of crude polysaccharides and total flavonoids are shown in Table 3.

[0148] Table 3 Detection results of crude polysaccharides and total flavonoids

[0149] Serial number Extraction time, h Total crude polysaccharides, mg Total flavonoids, mg 1 1 1440 53 2 1.5 1512 57 3 2.0 1404 49 4 2.5 1260 50 5 3.0 1368 48

[0150] (2) Results: Based on the comprehensive analysis of the contents of crude polysaccharides and total flavonoids, the optimal extraction time is 1.5 h.

[0151] 2.3 Screening of solid-liquid ratio for secondary extraction

[0152] 2.3.1. Material-liquid ratio group setting

[0153] The solid-liquid ratios (W / W or W / V) are 1:6, 1:7, 1:8, 1:9, and 1:10, respectively.

[0154] 2.3.2 Extraction method

[0155] Add water at a solid-liquid ratio of 1:10, raise the temperature to 45°C and soak for 1 hour, boil and extract for 1.5 hours, and then pass through a 100-mesh sieve to obtain a primary extract; then boil and extract for 1 hour at a solid-liquid ratio of 1:6, 1:7, 1:8, 1:9, 1:10, pass through a 100-mesh sieve to obtain a secondary extract, and combine the two extracts.

[0156] 2.3.3 Results

[0157] (1) The results of crude polysaccharides and total flavonoids detection are shown in Table 4.

[0158] Table 4 Detection results of crude polysaccharides and total flavonoids

[0159] Serial number Material-liquid ratio Total crude polysaccharides, mg Total flavonoids, mg 1 1:6 1382.4 70.4 2 1:7 1285.2 83.3 3 1:8 3045.6 81 4 1:9 1846.8 81.7 5 1:10 1008 72

[0160] (2) Results: Based on the comprehensive analysis of the final crude polysaccharide content and total flavonoids content of the two extractions, it was found that the optimal solid-liquid ratio for the secondary extraction was 1:8.

[0161] 2.4. Screening of extraction time for secondary extraction

[0162] 2.4.1. Extraction time group settings

[0163] The extraction times are 0.5h, 1.0h, 1.5h, 2.0h and 2.5h respectively.

[0164] 2.4.2 Extraction method

[0165] Add water at a solid-liquid ratio of 1:10, raise the temperature to 45°C and soak for 1 hour, boil and extract for 1.5 hours, then pass through a 100-mesh sieve to obtain a primary extract; then extract for 1 hour, 1.5 hours, 2.0 hours, 2.5 hours, and 3.0 hours at a solid-liquid ratio of 1:8, then pass through a 100-mesh sieve to obtain an extract, and combine the two filtrates.

[0166] 2.4.3 Results

[0167] The results of crude polysaccharides and total flavonoids detection are shown in Table 5.

[0168] Table 5 Detection results of crude polysaccharides and total flavonoids

[0169] Serial number Extraction time, h Total crude polysaccharides, mg Total flavonoids, mg 1 0.5 2008.8 72 2 1.0 2332.8 79.2 3 1.5 2008.8 82.8 4 2.0 1944 81 5 2.5 1814.4 88.2

[0170] (2) Results: Based on the comprehensive analysis of the final crude polysaccharide content and total flavonoids content of the two extractions, it was found that the optimal time for the secondary extraction was 1 h.

[0171] In summary, the optimal extraction process is: the solid-liquid ratio of the first extraction is 1:10, soaking at 45℃ for 1h, and boiling extraction time is 1.5h; the solid-liquid ratio of the second extraction is 1:8, and boiling extraction time is 1h.

[0172] Example 2 Strain Research

[0173] 1. Fermentation strain screening

[0174] 1.1. Single strain screening

[0175] Seven bacterial strains from the laboratory were initially selected: OD of each strain at the time of inoculation 600 As follows, Lactobacillus plantarum OD 600 3.0-5.0, Lactobacillus rhamnosus OD 600 3.5-5.5, Lactobacillus casei OD 600 3.5-5.0, Lactobacillus fermentans OD 600 3.5-5.0, Lactobacillus paracasei OD600 3.5-5.0, Lactobacillus delbrueckii subsp. bulgaricus OD 600 2.5-3.5 and Lactobacillus helveticus OD 600 3.5-5.0.

[0176] 1.2 Fermentation medium and conditions

[0177] 1.2.1 Fermentation medium: Chinese medicine extract with ginseng extract

[0178] 1.2.2 Fermentation conditions: single strain inoculation amount 3% (V / V), initial pH 6.5, 37°C, static fermentation for 120h, sampling and testing every 24h, pH and OD 600 .

[0179] 1.3 Results

[0180] 1.3.1. The test results of pH of fermentation broth are shown in Table 6.

[0181] Table 6 pH value of fermentation liquid of each strain at different fermentation time periods

[0182]

[0183] The lower the pH value in the fermentation process, the stronger the carbon source used by the strain, the better the decomposition ability of the effective components of Chinese medicine, and the more conducive to fermentation conversion. According to Table 6, it can be concluded that the fermentation time is 120h, and the pH value of the fermentation liquid of Lactobacillus delbrueckii subspecies bulgaricus and fermentation Lactobacillus mucilaginosus is above 4.0, which does not meet the fermentation requirements. The remaining 5 strains grow better, and from 48h, the pH value is below 4.0, which meets the fermentation requirements.

[0184] 1.3.2. Fermentation broth biological activity OD 600 The results of the value determination are shown in Table 7.

[0185] Table 7 Bioactivity OD of each strain at different fermentation stages 600 value

[0186]

[0187] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data, and the experimental data were expressed as mean + SD.

[0188] According to Table 7, OD in each time period of the fermentation process 600 The results showed that the fermentation broth of Lactobacillus delbrueckii subsp. bulgaricus had an OD value of 600The value is very low, indicating that the strain has basically no growth during the entire fermentation process and cannot meet the fermentation requirements. The remaining 6 strains were fermented for 48 hours, among which Lactobacillus plantarum, Lactobacillus rhamnosus and Lactobacillus helveticus grew better. 600 The values ​​all reached above 9.5, and reached above 10.2 after 120 hours of fermentation, indicating that the strain can make full use of the nutrients in the culture medium for growth and reproduction, and further indicating that the strain can effectively decompose or transform the traditional Chinese medicine components in the fermentation medium.

[0189] 1.4 Conclusion

[0190] In summary, according to the pH and OD of the fermentation process 600 The initial screening revealed that the fermentation strains were Lactobacillus plantarum, Lactobacillus rhamnosus and Lactobacillus helveticus.

[0191] 2. Strain combination research

[0192] Combined fermentation was carried out based on the strains obtained in the initial screening (Lactobacillus plantarum, Lactobacillus rhamnosus and Lactobacillus helveticus).

[0193] 2.1. Strain Combination

[0194] Three bacterial strains were used for combined fermentation experiments: No. 1: Lactobacillus plantarum and Lactobacillus rhamnosus; No. 2: Lactobacillus plantarum and Lactobacillus helveticus; No. 3: Lactobacillus rhamnosus and Lactobacillus helveticus; No. 4: Lactobacillus plantarum, Lactobacillus rhamnosus and Lactobacillus helveticus.

[0195] 2.2 Fermentation conditions

[0196] 2.2.1 Fermentation medium: Chinese medicine extract with ginseng extract

[0197] 2.2.2 Fermentation conditions: The experiment was carried out with a single strain 2% inoculation (V / V), the fermentation starting pH was 6.5, and the fermentation was allowed to stand at 37°C for 120 h. The pH and OD of the fermentation liquid were tested every 24 h. 600 and total flavonoids content to select the optimal combination of strains.

[0198] 2.3 Results

[0199] 2.3.1. The test results of pH of fermentation broth are shown in Table 8.

[0200] Table 8 pH value of fermentation broth of each strain combination at different fermentation time periods

[0201]

[0202] According to Table 8, during the fermentation process, the pH of the fermentation broth of the four strain combinations was below 4.0, with no significant difference.

[0203] 2.3.2 Fermentation broth biological activity OD 600 The results of the value determination are shown in Table 9.

[0204] Table 9 Bioactivity OD of each strain combination at different fermentation time periods 600 value

[0205]

[0206] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data, and the experimental data were expressed as mean + SD.

[0207] According to the results in Table 9, it can be concluded that the biological activity OD of the fermentation 24h4 strain combination of Lactobacillus plantarum, Lactobacillus rhamnosus and Lactobacillus helveticus 600 The value was low, indicating that the synergy between the three strains was poor in the early stage of fermentation; the OD 600 The values ​​were all very high, indicating good growth and good synergy between strains.

[0208] 2.3.3 The results of determination of total flavonoids content in fermentation broth are shown in Table 10.

[0209] Table 10 Total flavonoid content of fermentation broth of each bacterial strain combination at different fermentation time periods (unit: mg / mL)

[0210]

[0211] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD, and all experimental group data were analyzed by one-way ANOVA. P < 0.05 was statistically significant. The total flavonoids of other fermentation strain combinations were compared with combination 1, ***P < 0.001.

[0212] According to Table 10, at 24h and 48h of fermentation, the total flavonoid content of strain combination No. 1 was the highest and significantly different from other combinations (P < 0.001). The total flavonoid content of strain combination No. 2 during fermentation was the lowest, indicating that the synergy between Lactobacillus plantarum and Lactobacillus helveticus was poor.

[0213] Comprehensive fermentation broth pH, ​​biological activity OD 600 The values ​​and total flavonoids contents were determined, and the best fermentation strain combination was determined to be combination No. 1: Lactobacillus plantarum and Lactobacillus rhamnosus.

[0214] Example 3 Fermentation Study

[0215] 1. Study on the ratio of combined inoculation of strains

[0216] 1.1. Group settings

[0217] Inoculation ratio, fermentation conditions and detection indicators are shown in Table 11.

[0218] Table 11 Inoculation ratio, fermentation conditions and detection indicators

[0219]

[0220] 1.2 Results

[0221] 1.2.1. pH values ​​at different fermentation times, see Table 12.

[0222] Table 12 pH value at different fermentation time

[0223]

[0224] According to Table 12, the pH of the fermentation broth at different fermentation time periods was below 4.0, and there was basically no difference between the groups.

[0225] 1.2.2. Bioactivity OD at different fermentation times 600 For values, see Table 13.

[0226] Table 13 Bioactivity OD at different fermentation times 600 value

[0227]

[0228] Note: Graphpad Prism 9.5.1 software was used for statistical analysis of data, and the experimental data were expressed as mean + SD.

[0229] According to Table 13, the OD of the fermentation broth at different fermentation time periods 600 The values ​​were all very high, indicating that the strains at each inoculation ratio grew well.

[0230] 1.2.3 The total flavonoid content of the fermentation broth at different fermentation time periods is shown in Table 14.

[0231] Table 14 Total flavonoid content of fermentation broth at different fermentation time periods (unit: mg / mL)

[0232]

[0233] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD, and the experimental group data were analyzed by one-way ANOVA. P < 0.05 was statistically significant. The total flavonoids in the fermentation broth of the fermentation groups with other inoculation ratios were compared with the inoculation ratio of 1:1, ***P < 0.001.

[0234] According to Table 14:

[0235] (1) At 24h and 48h of fermentation, the total flavonoids content of the fermentation broth with an inoculation ratio of 1:1 was the highest and was significantly different from that of other combinations (P < 0.001).

[0236] (2) After 72 h of fermentation, the total flavonoid content of the fermentation liquid with an inoculation ratio of 1:1 decreased, while the total flavonoid content of the fermentation liquid with an inoculation ratio of 3:2 was superior to that of the other combinations.

[0237] Comprehensive fermentation broth pH, ​​biological activity OD 600 The optimal inoculation ratio of Lactobacillus plantarum and Lactobacillus rhamnosus was determined to be 1:1.

[0238] 2. Total inoculation volume study

[0239] 2.1. Group settings

[0240] The total inoculation amount, fermentation conditions and detection indicators are shown in Table 15.

[0241] Table 15 Total inoculation amount, fermentation conditions and detection indicators

[0242]

[0243]

[0244] 2.2 Results

[0245] 2.2.1. The pH values ​​at different fermentation times are shown in Table 16.

[0246] Table 16 pH value at different fermentation time

[0247]

[0248] According to Table 16, the pH of the fermentation broth at different fermentation time periods was below 4.0, and there was basically no difference between the groups.

[0249] 2.2.2. Bioactivity OD at different fermentation times 600 Values ​​are shown in Table 17.

[0250] Table 17 Bioactivity OD at different fermentation times 600 value

[0251]

[0252] Note: Graphpad Prism 9.5.1 software was used for statistical analysis of data, and the experimental data were expressed as mean + SD.

[0253] According to Table 17, the OD of the fermentation broth at different fermentation time periods 600 The values ​​were all very high, indicating that the strains at each inoculation ratio grew well.

[0254] 2.2.3. The total flavonoid content of the fermentation liquid at different fermentation time periods is shown in Table 18.

[0255] Table 18 Total flavonoid content of fermentation broth at different fermentation time periods (unit: mg / mL)

[0256]

[0257] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD, and all experimental group data were analyzed by one-way ANOVA. P < 0.05 was statistically significant. Compared with the total flavonoids in the fermentation broth of the fermentation group with the 4% inoculation amount, ***P < 0.001.

[0258] According to Table 18:

[0259] (1) After 24 hours of fermentation, the total flavonoids content in the fermentation broth with an inoculum size of 4% was the highest and significantly different from those with 2%, 3% and 5% inoculation (P < 0.001); there was no statistical difference in the total flavonoids content in the fermentation broth with an inoculum size of 6% (P > 0.05).

[0260] (2) After 48 h of fermentation, the total flavonoids content of the fermentation broth with an inoculum size of 4% was the highest and was significantly different from that of other combinations (P < 0.001).

[0261] (3) After 72h of fermentation, the total flavonoid content of the fermentation broth with 4% inoculation was higher than that of the fermentation broth with 2%, 5% and 6% inoculation, and the difference was extremely significant (P < 0.001). The total flavonoid content of the fermentation broth with 4% inoculation was lower than that of the fermentation broth with 3% inoculation, and the difference was extremely significant (P < 0.001).

[0262] Comprehensive fermentation broth pH, ​​biological activity OD 600 The values ​​and total flavonoids contents were determined and the total inoculum amount of Lactobacillus plantarum and Lactobacillus rhamnosus was 4%.

[0263] 3. Fermentation time study

[0264] 3.1. Group settings

[0265] Fermentation time, fermentation conditions and detection indicators are shown in Table 19.

[0266] Table 19 Fermentation time, fermentation conditions and detection indicators

[0267]

[0268] 3.2 Results

[0269] The results of total flavonoids determination are shown in Table 20.

[0270] Table 20 Total flavonoid content of fermentation broth at different fermentation time periods (mg / mL)

[0271]

[0272] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD, and the experimental group data were analyzed by one-way ANOVA. P < 0.05 was statistically significant. Compared with fermentation 32h, *P < 0.05 and ***P < 0.001 were found in other fermentation time periods.

[0273] According to Table 20:

[0274] (1) The total flavonoids content of the fermentation broth was the highest after fermentation for 24h, 32h and 48h.

[0275] (2) There was no statistical difference in the total flavonoids content of the fermentation broth after 24h and 48h compared with that of the fermentation broth after 32h (P>0.05). There was a statistical difference in the total flavonoids content of the fermentation broth after other fermentation times compared with that of the fermentation broth after 32h (P<0.05), and there was a very significant difference in the fermentation broth at other time periods (P<0.001).

[0276] In summary, in order to control production costs, it is better to control the fermentation time within 24 to 32 hours.

[0277] Example 4 Preparation method of extract, fermentation liquid and Chinese medicine composition compound fermented beverage

[0278] 1. Preparation of extract

[0279] Step 1: A Chinese medicine composition with medicinal and edible properties, wherein the weight components of Chinese medicinal materials are 75 parts of angelica, 75 parts of jujube, 75 parts of ginger, 50 parts of polygonatum, 50 parts of kudzu root, 50 parts of poria, 50 parts of dried orange peel, 25 parts of cinnamon, and 25 parts of licorice, which are mixed and packed in a Chinese medicine extraction filter bag, and the amount of each bag is about two-thirds of the bag. Add 12 times the volume ratio of drinking water to the Chinese medicine mixture for rinsing, rinse for 10 minutes, drain the rinsing water, add 12 times the volume ratio of drinking water for rinsing for 3 minutes, and drain the rinsing water. After rinsing, add 10 times the volume ratio of drinking water and soak at 45°C for 1 hour, then boil and extract for 1.5 hours, and filter through a double pipeline filter with 100 mesh; add 8 times the volume ratio of drinking water to the filter residue and boil and extract for 1 hour, filter through a double pipeline filter with 100 mesh, and combine the two filtrates.

[0280] Step 2: Stop concentrating the double-effect concentrate until the volume ratio reaches 3 times, add 25 parts of ginseng extract to the concentrate, and stir evenly to obtain the extract.

[0281] 2. Preparation of seed solution

[0282] (1) Primary seed liquid culture

[0283] 1) Prepare primary seed culture medium (MRS medium), 600 mL / bottle, 3 bottles, pH not controlled, sterilize at 115°C for 30 min.

[0284] 2) 1% (V / V) of Lactobacillus plantarum and 1% (V / V) of Lactobacillus rhamnosus were inoculated with Lactobacillus plantarum. 600 3.0~3.5, OD when inoculated with Lactobacillus rhamnosus 600 3.5~5.5, after thawing, inoculate into MRS liquid culture medium and culture in a 37℃ constant temperature incubator for 16~18h.

[0285] 3) Testing: Take a sample of the first-level seed liquid and test it under a microscope to ensure that there is no contamination by foreign bacteria. Test the OD of the seed liquid. 600 (Lactobacillus plantarum OD 600 3.0~5.0; Lactobacillus rhamnosus OD 600 3.5~5.5).

[0286] (2) Secondary seed liquid culture

[0287] 1) Sterilization of seed tanks: The seed tanks were first sterilized at 121°C for 30 min, and then the secondary culture medium (MRS culture medium) was prepared, 20 L / tank, 2 tanks, pH was not controlled, and sterilized at 115°C for 30 min.

[0288] 2) Seed tank inoculation: The cultured primary seed solution (inoculum amount: Lactobacillus plantarum 3% (V / V), Lactobacillus rhamnosus 3% (V / V)) was inoculated into two seed tanks respectively, and the fermentation parameters (temperature 37° C.; stirring speed 50 rpm; tank pressure 0.04-0.06 MPa) were maintained, and the pH was not controlled, and the culture was carried out for 10-14 h.

[0289] 3) Testing: Take samples of the secondary seed liquid for microscopic testing to ensure that there is no contamination by foreign bacteria and test the seed liquid OD 600 (Lactobacillus plantarum OD 600 3.0~5.0; Lactobacillus rhamnosus OD 600 3.5~5.5).

[0290] 3. Fermentation culture

[0291] 3.1. Sterilization of fermentation medium: The fermentation tank was emptied and sterilized at 121°C for 30 min in advance, and then the Chinese medicine extract concentrate was sterilized at 115°C for 0.5 h in a (300L) fermentation tank and cooled to 37°C.

[0292] 3.2. Fermentation inoculation: 3 groups of fermentation inoculations with different formulas

[0293] 3.2.1. Formula of the present invention: The cultured secondary seed solution (total inoculation amount is 4%: 2% of Lactobacillus plantarum and 2% of Lactobacillus rhamnosus, V / V) is inoculated into a fermentation tank.

[0294] 3.2.2. Comparative formula 1 (see Example 10 for the formula): The cultured secondary seed solution (total inoculation amount is 4%: Lactobacillus plantarum 2%, Lactobacillus rhamnosus 2%, V / V) is inoculated into a fermentation tank.

[0295] 3.2.3. Comparative formula 2 (see Example 10 for the formula): The cultured secondary seed solution (total inoculation amount is 4%: Lactobacillus plantarum 2%, Lactobacillus rhamnosus 2%, V / V) is inoculated into a fermentation tank.

[0296] 3.3. Adjust the fermentation conditions: the temperature is 37°C, the rotation speed is 50 rpm, the initial pH is 6.5, and the tank pressure is 0.04-0.06 MPa. Cultivate for 24-32 hours. Check the pH of the fermentation liquid every 8 hours. Fermentation is completed when the pH of the fermentation liquid is in the range of 3.3-3.8.

[0297] 3.4. After the fermentation is completed, sterilize at 72-75℃ for 0.5h, homogenize under high pressure for 0.5h, and filter through 10μm or use disc centrifugation to obtain the fermentation liquid.

[0298] 4. Preparation of compound fermented beverages of Chinese medicine composition

[0299] (1) Preparation of compound fermented beverages of Chinese medicine compositions

[0300] The preparation process is as follows: 3000 weight parts of the prepared fermentation broth (the fermentation broth is the fermentation broth obtained in the above step 3.4), adding seasoning materials 350 weight parts of oligofructose, 200 weight parts of erythritol, 100 weight parts of black wolfberry juice powder, 100 weight parts of xylitol, 10 weight parts of γ-aminobutyric acid, and 5 weight parts of taurine, and preparing the product through sugaring, filling, and sterilization, 50 weight parts / bag.

[0301] (2) Preparation of a Chinese medicine compound fermented beverage (without γ-aminobutyric acid)

[0302] The preparation process is as follows: 3000 weight parts of the prepared fermentation liquid (the fermentation liquid is the fermentation liquid obtained in the above step 3.4), adding seasoning materials 350 weight parts of oligofructose, 200 weight parts of erythritol, 100 weight parts of black wolfberry juice powder, 100 weight parts of xylitol, and 5 weight parts of taurine, and preparing it through sugaring, filling, and sterilization, 50 weight parts / bag.

[0303] Example 5 Study on Antioxidant Effect

[0304] 1. Determination of DPPH free radical scavenging rate

[0305] 1.1 Experimental Purpose

[0306] Investigate the DPPH radical scavenging rate of the extract and fermentation broth obtained in Example 4

[0307] 1.2 Research Content

[0308] 1.2.1. Preparation of experimental reagents

[0309] 0.1mmol / L DPPH solution: Accurately weigh 0.004g of DPPH into a beaker, add a small amount of anhydrous ethanol to fully dissolve it, place it in a 100mL volumetric flask and dilute to the mark with anhydrous ethanol to prepare a 0.1mmol / L DPPH solution. The solution should be used immediately after preparation and stored away from light.

[0310] 1.2.2 Experimental steps

[0311] 1.2.2.1. Extraction solution determination

[0312] Sample group A1: Take 2 mL of the extract and 2 mL of the DPPH solution in a test tube and shake them thoroughly to mix them evenly;

[0313] Control group A2: Take 2 mL of the extract and 2 mL of anhydrous ethanol in a test tube and shake them thoroughly to mix them evenly;

[0314] Blank group A3: Take 2 mL of ultrapure water and 2 mL of DPPH solution in a test tube and shake them thoroughly to mix them evenly;

[0315] A1, A2 and A3 were reacted at room temperature in the dark for 30 min, and anhydrous ethanol was used as a blank control. The absorbance was measured at 517 nm, and three parallel experiments were performed.

[0316] The calculation formula is as follows:

[0317]

[0318] 1.2.2.2. Fermentation broth determination

[0319] The determination method is the same as that of the extract solution in 1.2.2.1.

[0320] 1.3 Test results

[0321] The test results are shown in Table 21.

[0322] Table 21 Comparative test of DPPH free radical scavenging rate of extract and fermentation broth

[0323]

[0324] Note: Graphpad Prism 9.5.1 software was used for statistical analysis of the data. The experimental data were expressed as mean + SD. The t-test was used between the extract group and the fermentation liquid group, with the test level of α = 0.05, p < 0.05 indicated a statistical difference, **p < 0.01 indicated a significant difference.

[0325] The results in Table 21 show that there is a significant difference in the DPPH free radical scavenging rate between the fermentation liquid group and the extract liquid group (p < 0.01), and the antioxidant effect is better after fermentation.

[0326] 2. Determination of hydroxyl radical scavenging rate

[0327] 2.1 Experimental Purpose

[0328] The hydroxyl radical scavenging rates of the extract and fermentation broth prepared in Example 4 were investigated.

[0329] 2.2 Research Content

[0330] 2.2.1. Preparation of experimental reagents

[0331] (1) 9 mmol / L ferrous sulfate solution: Accurately weigh 0.25 g of ferrous sulfate into a beaker, dissolve it fully with a small amount of purified water, and place it in a 100 mL volumetric flask to the mark.

[0332] (2) 9 mmol / L salicylic acid-ethanol solution: Accurately weigh 0.1243 g of salicylic acid in a beaker, dissolve it fully in a small amount of anhydrous ethanol, and place it in a 100 mL volumetric flask and dilute to the mark.

[0333] (3) 8.8mmol / L H2O2 solution: take 1mL of 30% H2O2 and add purified water to make up to 1L.

[0334] 2.2.2 Experimental steps

[0335] 2.2.2.1. Extraction solution determination

[0336] Sample group A1: Take 1 mL of the extract in a test tube, add 1 mL of 9 mmol / L ferrous sulfate solution and 1 mL of 9 mmol / L salicylic acid-ethanol solution in sequence, and shake well to mix them evenly; then add 1 mL of 8.8 mmol / L H2O2 solution to start the reaction, and shake well to mix them evenly.

[0337] Control group A2: Take 1 mL of the extract in a test tube, add 1 mL of 9 mmol / L ferrous sulfate solution and 1 mL of 9 mmol / L salicylic acid-ethanol solution in sequence, and shake well to mix them evenly; then add 1 mL of ultrapure water and shake well to mix them evenly.

[0338] Blank group A3: Take 1 mL of ultrapure water in a test tube, add 1 mL of 9 mmol / L ferrous sulfate solution and 1 mL of 9 mmol / L salicylic acid-ethanol solution in sequence, and shake well to mix them evenly; then add 1 mL of 8.8 mmol / L H2O2 solution to start the reaction, and shake well to mix them evenly.

[0339] A1, A2 and A3 were placed in a constant temperature water bath at 37°C for reaction for 30 min. Ultrapure water was used as a blank control. The absorbance was measured at 510 nm. Three parallel experiments were performed.

[0340] The calculation formula is as follows:

[0341]

[0342] 2.2.2.2. Fermentation broth determination

[0343] The determination method is the same as that of the extract in 2.2.2.1

[0344] 2.3 Experimental Results

[0345] The experimental results are shown in Table 22.

[0346] Table 22 Comparative test of hydroxyl radical scavenging rate of extract and fermentation broth

[0347]

[0348] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of the data. The experimental data were expressed as mean + SD. The t-test was used between the extract group and the fermentation liquid group, with the test level of α = 0.05, p < 0.05 indicated statistical difference, and ***p < 0.001 indicated extremely significant difference.

[0349] The results in Table 22 show that there is a significant difference in the hydroxyl radical scavenging rate between the fermentation liquid group and the extract liquid group (p < 0.01), and the antioxidant effect is better after fermentation.

[0350] 3. Total antioxidant assay

[0351] 3.1 Experimental Purpose

[0352] 3.2.1. Preparation of experimental reagents

[0353] Ammonium molybdate solution: accurately weigh 0.99 g of ammonium molybdate tetrahydrate, 2.13 g of sodium phosphate, and 6.67 mL of concentrated sulfuric acid, add deionized water to a 200 mL volumetric flask to make the concentrations 4 mmol / L, 28 mmol / L, and 0.6 mol / L, respectively, mix well to make an ammonium molybdate solution for use.

[0354] 3.2.2 Experimental steps

[0355] 3.2.2.1. Extraction solution determination

[0356] Take 1mL of the extract in a test tube, add 3.0mL of the prepared ammonium molybdate solution, mix well, seal and place in a constant temperature water bath at 95℃ for 90min, take it out and quickly cool it to room temperature, use distilled water as a blank, and measure the absorbance at 695nm. The larger the absorbance value, the stronger the antioxidant capacity.

[0357] Blank control group: Replace the sample solution with an equal volume of deionized water as a blank control, and the method is the same as above.

[0358] The calculation formula is as follows:

[0359] Total antioxidant capacity OD = sample OD value ~ blank control OD value

[0360] 3.2.2.2 Fermentation broth determination

[0361] The determination method is the same as that of the extract in 3.2.2.1

[0362] 3.3 Results

[0363] The experimental results are shown in Table 23.

[0364] Table 23 Comparative test of total antioxidant activity of extract and fermentation broth

[0365]

[0366] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of the data. The experimental data were expressed as mean + SD. The t-test was used between the extract group and the fermentation liquid group, with the test level of α = 0.05, p < 0.05 indicated statistical difference, and ***p < 0.001 indicated extremely significant difference.

[0367] The results in Table 23 show that the total antioxidant activity between the fermentation liquid group and the extract liquid group has a very significant difference (p < 0.001), and the antioxidant effect is better after fermentation.

[0368] Based on the results of the DPPH radical scavenging rate, hydroxyl radical scavenging rate and total antioxidant activity of the above-mentioned extract and fermentation liquid, it is shown that the weight components of the traditional Chinese medicine provided by the present invention and the fermentation treatment of the extract using mixed bacteria can improve the antioxidant activity of the product.

[0369] Example 6 Cell Efficacy Study

[0370] Test drugs: the extract and fermentation broth prepared in Example 4.

[0371] 1. Cell safety study

[0372] 1.1. Cell safety studies

[0373] 1.1.1. Raw264.7 cell safety experimental methods and procedures

[0374] (1) Plating: Inoculate cells: Prepare a single cell suspension of Raw264.7 cells (purchased from the Cell Bank of the Chinese Academy of Sciences in November 2017) using 1640 complete culture medium and adjust the cell density to 10 cells per well. 5 Cells were inoculated into 96-well plates, with a volume of 100 μL per well, and 100 μL of culture medium was added. They were placed in an incubator and cultured overnight. Three parallel wells were set up for each group.

[0375] (2) Sample addition treatment: Place the cells in a CO2 incubator (containing 5% carbon dioxide gas by volume, the same below) at 37°C for 24 hours, then discard the original culture medium. Add 200 μL of the sample group solution and continue culturing for 24 hours. The blank group is cells cultured in normal cell culture medium.

[0376] (3) Adding MTT: 24 hours after replacing the culture medium, observe the cell morphology under a microscope. Add 20 μL of MTT solution (5 mg / mL, i.e., 0.5% MTT) to each well and continue culturing for 4 hours. Aspirate the supernatant, add 150 μL of DMSO, and oscillate on an oscillator until the crystals are completely dissolved. Measure the absorbance on a microplate reader with a main absorption wavelength of 570 nm.

[0377] 1.2. Raw264.7 cell safety results, see Table 24.

[0378] Table 24 Cell safety test results

[0379]

[0380] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD. The t-test was used between the extract group and the fermentation liquid group. The test level was α = 0.05, and ***p < 0.001 indicated a very significant difference. Cell death rate > 80% was considered non-toxic.

[0381] The results in Table 24 show that:

[0382] (1) The cell safety of both the extract and fermentation broth groups was >80%, indicating good safety;

[0383] (2) The cell safety of the fermentation liquid group was better than that of the extract group (p < 0.001), and the fermented product had a better nourishing effect on cells.

[0384] 2. Cell phagocytosis index study

[0385] 2.1. Experimental methods and steps of phagocytic index of mouse macrophages (Raw264.7 cells)

[0386] (1) The cultured cells were resuspended in DMEM high-glucose medium (containing 10% FBS + 1% double antibody) and inoculated into a 96-well plate. The cell density in the 96-well plate was 3×10 4 The cells were cultured in a carbon dioxide incubator for 12 h.

[0387] (2) The supernatant was discarded, and H2O2 solution was added for 24 h. Then, 800 μmol / L hydrogen peroxide was added to the model group, the extraction liquid group, and the fermentation liquid group for 24 h. The supernatant was discarded, and 200 μL DMEM high-glucose complete medium was added to the blank group and the model group. The samples of the extraction group and the fermentation group were diluted 25 times with DMEM high-glucose complete medium and 200 μL was added to a 96-well plate and incubated for 24 h.

[0388] (3) Add 20 μL of neutral red staining solution and incubate in a cell culture incubator for 4 h. Remove the cell culture medium containing neutral red staining solution and wash with PBS solution 1 to 2 times. Add 200 μL of neutral red detection lysis solution and lyse on a shaker at room temperature for 10 min. Measure the absorbance at A540 nm.

[0389] The calculation formula is as follows:

[0390]

[0391] 2.2 Results

[0392] The results of the cell phagocytic index experiment are shown in Table 25.

[0393] Table 25 Cell phagocytosis index experimental results

[0394] Blank group Model Group Extract group Fermentation broth group 1.000 0.799±0.006### 0.853±0.009 0.937±0.014***

[0395] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD, and the experimental group data were analyzed by one-way ANOVA. P < 0.05 was statistically significant, ###P < 0.001 compared with the blank group, and ***P < 0.001 compared with the fermentation liquid group and the extract group.

[0396] The results in Table 25 show

[0397] (1) There was a significant difference between the model group and the blank group (p < 0.001);

[0398] (2) Compared with the model group, both the extract group and the fermentation liquid group could increase the phagocytic index of cells; there was a very significant difference between the fermentation liquid group and the extract (p < 0.001), indicating that the fermentation liquid group had a higher ability to improve immunity than the extract group.

[0399] 3. Cellular superoxide dismutase (SOD) activity detection experiment

[0400] 3.1. Raw264.7 Cell SOD Experiment Methods and Steps

[0401] (1) RAW264.7 cells in the logarithmic growth phase were inoculated into 3 cm culture dishes at a cell density of 1 × 10 7 / mL, the inoculation volume was 1mL per well, and the culture was carried out for 12h.

[0402] (2) Except for the blank control group, 3 mL of 800 μmol / L H2O2 solution was added to the model group, extract group, and fermentation liquid group, and the culture dish was transferred to a cell culture incubator at 37°C and 5% CO2 for 24 h, and the supernatant was discarded. Then 3 mL of DMEM high-glucose medium was added to the blank control group and the model group, 3 mL of 25-fold diluted extract was added to the extract group, and 3 mL of 25-fold diluted fermentation liquid was added to the fermentation liquid group, and the culture dish was transferred to a cell culture incubator at 37°C and 5% CO2 for 24 h.

[0403] (3) After the intervention, wash with PBS 1-2 times, centrifuge at 5000rpm for 3min, collect the precipitate, dissolve it with 50μL SOD extract, and transfer it to a culture dish. Then add 150μL SOD extract to each dish, scrape all the cells in each well with a cell scraper, collect them in a centrifuge tube, and ultrasonically disrupt them (power 20% or 200w, ultrasonic 3S, interval 10S, repeat 30 times). Centrifuge at 12000rpm for 10min, and take the supernatant for later use. Refer to the superoxide dismutase (SOD) activity detection kit (Solebo Biochemical Kit Division, product number BC0175) for detection.

[0404] 3.2 Results

[0405] The results of the cell SOD activity experiment are shown in Table 26.

[0406] Table 26 Cell SOD activity detection experimental results (U / mg port)

[0407] Blank group Model Group Extract group Fermentation broth group 4.909±0.011 0.197±0.014### 2.359±0.015 4.394±0.012***

[0408] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD, and the experimental group data were analyzed by one-way ANOVA. P < 0.05 was statistically significant, ###P < 0.001 compared with the blank group, and ***P < 0.001 compared with the fermentation liquid group and the extract group.

[0409] The results in Table 25 show that:

[0410] (1) There was a significant difference between the model group and the blank group (p < 0.001);

[0411] (2) Compared with the model group, both the extract group and the fermentation liquid group had stronger SOD activity; there was a very significant difference between the fermentation liquid group and the extract (p < 0.001), indicating that the activity of the fermentation liquid was better than that of the extract group.

[0412] Example 7 Study on the Anti-fatigue Effect of Extract, Fermentation Broth and Fermented Beverage on Zebrafish

[0413] 1. Study on the behavioral analysis of zebrafish fatigue model using extract and fermentation liquid

[0414] 1.1. Drugs under investigation

[0415] The extract and fermentation broth prepared in Example 4.

[0416] 1.2 Experimental Animals

[0417] Wild-type AB strain zebrafish, aged 5 df (5 days after fertilization), were provided by Qingdao Kangmaichen Biotechnology Co., Ltd.

[0418] 1.3 Main Reagents

[0419] Anhydrous sodium sulfite: anhydrous sodium sulfite + standard dilution water = 100 mg / mL, 240 μL → 3 mL, i.e. final concentration 8 mg / mL, prepare and use immediately.

[0420] 1.4 Experimental methods and steps

[0421] (1) 24h treatment

[0422] Blank group: Model zebrafish were fed with fish water (standard water); model group was fed with 8 mg / mL anhydrous sodium sulfite; extract group was fed with 8 mg / mL anhydrous sodium sulfite + 25 μL extract; fermentation group was fed with 8 mg / mL anhydrous sodium sulfite + 25 μL fermentation. Wrapped in aluminum foil and placed in a 28°C incubator for 24 hours.

[0423] (2) Modeling

[0424] First, deduct 240μL of fish water, then add 240μL of 100mg / mL anhydrous sodium sulfite. Observe 30 minutes after adding anhydrous sodium sulfite (the lid cannot be opened). When the zebrafish in the model control group lean against the wall and cannot swim to the middle, it proves that the model is successfully built. The time for successful modeling is not fixed, about 45min to 1.5h, and needs to be determined according to each situation.

[0425] (3) Movement distance observation

[0426] The fish were immediately transferred to a 96-well plate, 200 μL / tail / well, and the total movement distance of the zebrafish within 1 h was detected using a zebrafish behavior analyzer at 28°C in the dark.

[0427] 1.5 Experimental Results

[0428] The behavioral analysis results of the zebrafish fatigue model are shown in Table 27.

[0429] Table 27 Behavioral analysis results of zebrafish fatigue model in the extract group and fermentation liquid group (mm)

[0430] Blank group Model Group Extract group Fermentation broth group 5000.00±150.9 2140±138.2### 5612±180.2 7915±198.3***

[0431] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD, and the experimental group data were analyzed by one-way ANOVA. P < 0.05 was statistically significant, ###P < 0.001 compared with the model group and the blank group, and ***P < 0.001 compared with the extract group and the fermentation liquid group.

[0432] From Table 27, we can see that:

[0433] (1) There was a significant difference between the model group and the blank group (p < 0.001);

[0434] (2) The extract group increased the movement distance of zebrafish in the model group from 2140 mm to 5612 mm in the same period of time, an increase of 162.24%;

[0435] (3) In the same period of time, the fermentation liquid group increased the movement distance of zebrafish in the model group from 2140 mm to 7915 mm, an increase of 269.86%;

[0436] (4) There was a significant difference in the movement distance of zebrafish in the fermentation liquid group and the extract liquid group, ***P<0.001.

[0437] In summary, the anti-fatigue effect of the fermentation liquid group was better than that of the extract group.

[0438] 2. Research on the anti-fatigue effect of fermented drinks

[0439] 2.1. Drugs under investigation

[0440] The Chinese medicine composition compound fermented beverage prepared in Example 4 (2 bags / day), the Chinese medicine composition compound fermented beverage (without γ-aminobutyric acid, 2 bags / day), Bazhen Liquid (LSPG Guangdong Wannianqing Pharmaceutical Co., Ltd., batch number: 210710; 2 bottles / day), and marketed health food (Rijiaman Beverage (Shanghai) Co., Ltd., batch number: 2022011211; 1 bottle / day).

[0441] 2.2 Experimental Methods

[0442] The experimental methods and steps are the same as those in Section 1.4 of this embodiment.

[0443] 2.3 Results

[0444] The experimental results of zebrafish fatigue model behavior analysis are shown in Table 28.

[0445] Table 28 Zebrafish fatigue model behavior analysis experimental results (mm)

[0446]

[0447] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD, and the experimental group data were analyzed by one-way ANOVA. P < 0.05 was statistically significant, the model group was compared with the blank group ###P < 0.001, and the Chinese medicine composition compound fermented beverage (without γ-aminobutyric acid), Bazhen liquid and marketed health foods were compared with the Chinese medicine composition compound fermented beverage ***P < 0.001.

[0448] From Table 28, we can see that:

[0449] (1) There was a significant difference between the model group and the blank group (p < 0.001);

[0450] (2) There was a very significant difference between the Chinese medicine composition compound fermented beverage and the Chinese medicine composition compound fermented beverage (without γ-aminobutyric acid), Bazhen Liquid and marketed health foods, P < 0.001.

[0451] Based on the above results, it can be concluded that the Chinese medicine composition compound fermented beverage provided by the present invention has good anti-fatigue effect.

[0452] Example 8 Study on the anti-fatigue efficacy of extracts, fermented broths and fermented beverages in zebrafish

[0453] 1. The test drugs and experimental animals are the same as those in Example 7.

[0454] 2. Experimental methods and steps

[0455] (1) Fish selection: Select 5 df (5 days after fertilization) juvenile fish and add them to a 6-well plate, 80 per well. Aspirate the fish culture water and add prepared standard water (standard water preparation: accurately weigh 294.0 mg anhydrous calcium chloride, 123.3 mg magnesium sulfate heptahydrate, 63.0 mg sodium bicarbonate, and 5.5 mg potassium chloride, fully dissolve them in part of deionized water, and finally adjust the volume to 1 L), 9 mL per well.

[0456] (2) Dosing: Blank group: Model zebrafish were fed with fish water (standard water); model group was fed with 8 mg / mL anhydrous sodium sulfite; extract group was fed with 8 mg / mL anhydrous sodium sulfite + 25 μL extract; fermentation group was fed with 8 mg / mL anhydrous sodium sulfite + 25 μL fermentation; Chinese medicine compound fermented beverage group was fed with 8 mg / mL anhydrous sodium sulfite + 25 μL Chinese medicine compound fermented beverage; Chinese medicine compound fermented beverage (without γ-aminobutyric acid) group was fed with 8 mg / mL anhydrous sodium sulfite + 25 μL Chinese medicine compound fermented beverage (without γ-aminobutyric acid); Bazhen liquid group was fed with 8 mg / mL anhydrous sodium sulfite + 10 μL Bazhen liquid; marketed health food group was fed with 8 mg / mL anhydrous sodium sulfite + 10 μL marketed health food. Wrap with aluminum foil and place in a 28°C incubator for 24 h.

[0457] (3) Modeling: Anhydrous sodium sulfite was added to the model group and the experimental group to a final concentration of 8 mg / mL. Observation began 30 minutes after the addition of anhydrous sodium sulfite (the lid could not be opened). When the zebrafish in the model control group leaned against the wall and could not swim to the middle, the model was successfully built. The time for successful modeling was not fixed, ranging from about 45 minutes to 1.5 hours, and needed to be determined based on each situation.

[0458] (4) Pre-test treatment: The experimental fish were rinsed with pure water for 2 to 3 times. The fish of each experimental group were transferred to a 2 mL centrifuge tube, and the water was removed as much as possible. The extract was added and the tubes were ground at low temperature on a grinder and then centrifuged at 4°C for 10 min. The tubes were placed on ice for testing.

[0459] (5) Detection: Detection was performed according to the instructions of the lactic acid content detection kit (Solebo Biochemical Kit Division, product number BC2235).

[0460] 3. Experimental results

[0461] The lactic acid determination results are shown in Table 29.

[0462] Table 29 Lactic acid determination results (μmol / mgport)

[0463]

[0464]

[0465] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD, and the experimental group data were analyzed by one-way ANOVA. P < 0.05 was statistically significant. The comparison between the model group and the blank group was ###P < 0.001. The comparison between the model group, the extract group and the fermentation liquid group was **P < 0.01, ***P < 0.001. The comparison between the model group, the Chinese medicine compound fermented beverage (without γ-aminobutyric acid), Bazhen liquid and the marketed health food and the Chinese medicine compound fermented beverage was ΔΔΔP < 0.001.

[0466] From Table 29, we can see that:

[0467] (1) There was a significant difference between the model group and the blank group (p<0.001).

[0468] (2) There was a very significant difference between the model group and the fermentation liquid group (p < 0.001); there was a significant difference between the extract group and the fermentation liquid group (p < 0.01);

[0469] (3) There was no statistical difference between the Chinese medicine composition compound fermented beverage (without γ-aminobutyric acid) group and the Bazhenye group and the Chinese medicine composition compound fermented beverage group (p>0.05); there was a very significant difference between the model group and the marketed health food group and the Chinese medicine composition compound fermented beverage group (p<0.001).

[0470] Based on the above results, we can conclude

[0471] (1) The fermentation broth is more effective than the extract in reducing lactic acid content.

[0472] (2) The efficacy of the Chinese herbal medicine compound fermented beverage in reducing lactic acid content is no different from that of the Chinese herbal medicine compound fermented beverage (without γ-aminobutyric acid) and Bazhen Liquid, but is better than the health foods already on the market.

[0473] Example 9 Study on the anti-fatigue efficacy of extracts, fermented broths and fermented beverages in zebrafish

[0474] 1. The test drugs and experimental animals are the same as those in Example 7.

[0475] 2. Experimental methods and steps

[0476] The experimental method and steps are the same as those in Section 2 of Example 8. The detection is performed according to the instructions of the ATP content detection kit (Solar Biochemical Kit Division, product number BC0305).

[0477] 3. Experimental results

[0478] The ATP determination results are shown in Table 30.

[0479] Table 30 ATP determination results (μmol / g)

[0480]

[0481] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD, and the experimental group data were analyzed by one-way ANOVA. P < 0.05 was statistically significant. The comparison between the model group and the blank group was ###P < 0.001, the comparison between the model group, the extract group and the fermentation liquid group was ***P < 0.001, and the comparison between the model group, the Chinese medicine compound fermented drink (without γ-aminobutyric acid) and Bazhen Liquid and the Chinese medicine compound fermented drink was ΔΔΔP < 0.001.

[0482] From Table 30, we can see that:

[0483] (1) There was a significant difference between the model group and the blank group (p<0.001).

[0484] (2) There was a very significant difference between the model group and the fermentation liquid group (p < 0.001); there was a significant difference between the extract group and the fermentation liquid group (p < 0.001);

[0485] (3) The model group, the Chinese medicine compound fermented beverage (without γ-aminobutyric acid) group, and the Bazhenye group showed extremely significant differences compared with the Chinese medicine compound fermented beverage (p < 0.001).

[0486] Based on the above results, we can conclude

[0487] (1) The fermentation broth is more effective than the extract in increasing ATP content.

[0488] (2) The effect of the Chinese herbal medicine compound fermented beverage on increasing ATP content is better than that of Bazhenye, but lower than that of the Chinese herbal medicine compound fermented beverage (without γ-aminobutyric acid).

[0489] Example 10 Comparative experiment on the antioxidant efficacy of different formulations in vitro

[0490] 1. Formulation and preparation method

[0491] The formula of the present invention includes 75 parts of angelica, 75 parts of jujube, 75 parts of ginger, 50 parts of polygonatum, 50 parts of kudzu root, 50 parts of tuckahoe, 50 parts of tangerine peel, 25 parts of cinnamon bark and 25 parts of liquorice;

[0492] Extract preparation process (same as Example 4): Weigh the above medicinal materials for extraction, concentration, and add 25 parts of ginseng extract to the concentrate.

[0493] Fermentation broth preparation process (same as Example 4): The extract is fermented by composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or centrifuged to obtain a fermentation broth.

[0494] Comparative formula 1: 150 parts of angelica, 150 parts of jujube, 150 parts of ginger, 100 parts of polygonatum, 100 parts of kudzu root, 100 parts of tuckahoe, 100 parts of tangerine peel, 50 parts of cinnamon, and 50 parts of liquorice;

[0495] Extract preparation process (same as Example 4): Weigh the above medicinal materials for extraction, concentration, and add 37 parts of ginseng extract to the concentrate.

[0496] Fermentation broth preparation process (same as Example 4): The extract is fermented by composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or centrifuged to obtain a fermentation broth.

[0497] Comparative formula 2: 150 parts of angelica, 150 parts of jujube, 150 parts of ginger, 150 parts of polygonatum, 200 parts of kudzu root, 150 parts of tuckahoe, 150 parts of tangerine peel, 50 parts of cinnamon, and 50 parts of liquorice;

[0498] Extract preparation process (same as Example 4): Weigh the above medicinal materials for extraction, concentration, and add 37 parts of ginseng extract to the concentrate.

[0499] Fermentation broth preparation process (same as Example 4): The extract is fermented by composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or centrifuged to obtain a fermentation broth.

[0500] 2. Determination of DPPH free radical scavenging rate

[0501] 2.1 Methods and steps

[0502] The determination method is the same as the DPPH free radical scavenging rate determination method in Example 5.

[0503] 2.2 Results

[0504] 2.2.1. Comparative study of extracts with different formulas

[0505] The results of DPPH free radical scavenging rate determination are shown in Table 31.

[0506] Table 31 Comparative test of DPPH free radical scavenging rate of fermentation broth with different formulas

[0507]

[0508] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of the data. The experimental data were expressed as mean + SD. The t-test was used between the formula of the present invention and the comparative formula 1, with the test level α = 0.05, and ***p < 0.001 indicating a significant difference. The t-test was used between the formula of the present invention and the comparative formula 2, with the test level α = 0.05, and ###p < 0.001 indicating a significant difference.

[0509] As can be seen from Table 31, the DPPH radical scavenging rate of the extract of the formula of the present invention is lower than that of comparative formulas 1 and 2, and there is a very significant difference, indicating that increasing the amount of medicinal materials in the formula can significantly improve the antioxidant capacity of the extract.

[0510] 2.2.2 Comparative study of fermentation broths with different formulations

[0511] The results of DPPH free radical scavenging rate determination are shown in Table 32.

[0512] Table 32 Comparative test of DPPH free radical scavenging rate of fermentation broth with different formulas

[0513]

[0514] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD. A t-test was used between the present invention formula and comparative formula 1. *p < 0.05 indicated that there was a statistical difference between the groups. A t-test was used between the present invention formula and comparative formula 2. #p < 0.05 indicated that there was a statistical difference between the groups. The test level α = 0.05.

[0515] As shown in Table 32, the DPPH radical scavenging rate of the fermentation broth of the formula of the present invention is higher than that of comparative formulas 1 and 2, and is statistically different from the fermentation broth of comparative formula 1, p < 0.05; and is not statistically different from the fermentation broth of comparative formula 2, p > 0.05.

[0516] Based on the above results, it can be concluded that the amount of medicinal materials used in comparative formula 1 and comparative formula 2 is more than twice that of the formula of the present invention, and the DPPH free radical scavenging rate of the extracts is significantly higher than that of the formula of the present invention. However, after being treated by the same fermentation process, the DPPH free radical scavenging rate of the formula of the present invention is the highest and has a statistical difference with comparative formula 1, p < 0.05. Therefore, it is shown that the weight components of the Chinese medicinal materials provided by the present invention and the fermentation treatment of the extracts thereof using mixed bacteria can improve the antioxidant activity of the product, significantly save the amount of medicinal materials used, and reduce production costs.

[0517] 3. Comparative study on the anti-fatigue effect of zebrafish

[0518] 3.1 Methods and steps

[0519] The methods and steps are the same as those in Section 1.4 of Example 7.

[0520] 3.2 Results

[0521] The results of zebrafish movement distance detection are shown in Table 33.

[0522] Table 33 Comparative study results of zebrafish anti-fatigue efficacy (mm)

[0523] Blank group Model Group Formula of the present invention Comparison formula 1 3506±127.8 1023±123.7### 1958±134.1 1534±126.2***

[0524] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. The experimental data were expressed as mean + SD, and the experimental group data were analyzed by one-way ANOVA. P < 0.05 was statistically significant, the model group was compared with the blank group ###P < 0.001, and the comparison between the comparative formula 1 and the formula of the present invention was ***P < 0.001.

[0525] From Table 33, we can see that:

[0526] (1) There was a significant difference between the model group and the blank group (p < 0.001);

[0527] (2) There was a very significant difference in the movement distance of zebrafish of the present invention formula and comparative formula 1, ***P<0.001.

[0528] In summary, the weight components of the traditional Chinese medicinal materials preferably obtained in the present invention and the fermentation treatment of the extract thereof using mixed bacteria can significantly improve the anti-fatigue effect of the product.

[0529] Example 11 Comparative Experiment 1 on Cell Efficacy of Different Formulas

[0530] 1.1. The experimental cells, consumables, reagents, instruments, methods and steps are the same as those in Example 6.

[0531] 1.2. Formulation and preparation method

[0532] The formula of experimental group 1: 75 parts of angelica, 75 parts of jujube, 75 parts of ginger, 50 parts of polygonatum, 50 parts of kudzu root, 50 parts of poria, 50 parts of dried tangerine peel, 25 parts of cinnamon bark, and 25 parts of liquorice;

[0533] Extract preparation process (same as Example 4, the extract preparation process of the following formulas is the same as this formula): weigh the above medicinal materials for extraction, concentration, and add 25 parts of ginseng extract to the concentrate.

[0534] Fermentation broth preparation process (same as Example 4, the following fermentation broth preparation processes are the same as this formula): the extract is fermented with composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or disc centrifuged to obtain a fermentation broth.

[0535] The formula of experimental group 2: 75 parts of angelica, 75 parts of jujube, 30 parts of ginger, 50 parts of polygonatum, 50 parts of kudzu root, 50 parts of poria, 50 parts of dried tangerine peel, 25 parts of cinnamon, and 25 parts of liquorice;

[0536] Extract preparation process: weigh the above medicinal materials, extract, concentrate, and add 25 parts of ginseng extract to the concentrate.

[0537] Fermentation broth preparation process: the extract is fermented by composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or disc centrifuged to obtain the fermentation broth.

[0538] The formula of experimental group 3: 75 parts of angelica, 75 parts of jujube, 140 parts of ginger, 50 parts of polygonatum, 50 parts of kudzu root, 50 parts of poria, 50 parts of dried tangerine peel, 25 parts of cinnamon bark, and 25 parts of liquorice;

[0539] Extract preparation process: weigh the above medicinal materials, extract, concentrate, and add 25 parts of ginseng extract to the concentrate.

[0540] Fermentation broth preparation process: the extract is fermented by composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or disc centrifuged to obtain the fermentation broth.

[0541] The formula of control group 1: 75 parts of angelica, 75 parts of jujube, 5 parts of ginger, 50 parts of polygonatum, 50 parts of kudzu root, 50 parts of tuckahoe, 50 parts of tangerine peel, 25 parts of cinnamon, and 25 parts of liquorice;

[0542] Extract preparation process: weigh the above medicinal materials, extract, concentrate, and add 25 parts of ginseng extract to the concentrate.

[0543] Fermentation broth preparation process: the extract is fermented by composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or disc centrifuged to obtain the fermentation broth.

[0544] The formula of control group 2: 75 parts of angelica, 75 parts of jujube, 9 parts of ginger, 50 parts of polygonatum, 50 parts of kudzu root, 50 parts of tuckahoe, 50 parts of tangerine peel, 25 parts of cinnamon bark, and 25 parts of liquorice;

[0545] Extract preparation process: weigh the above medicinal materials, extract, concentrate, and add 25 parts of ginseng extract to the concentrate.

[0546] Fermentation broth preparation process: the extract is fermented by composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or disc centrifuged to obtain the fermentation broth.

[0547] The formula of control group 3: 75 parts of angelica, 75 parts of jujube, 9 parts of ginger, 50 parts of kudzu root, 50 parts of poria, 25 parts of cinnamon bark, and 25 parts of liquorice;

[0548] Extract preparation process: weigh the above medicinal materials, extract, concentrate, and add 25 parts of ginseng extract to the concentrate.

[0549] Fermentation broth preparation process: the extract is fermented by composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or disc centrifuged to obtain the fermentation broth.

[0550] The formula of control group 4: 75 parts of angelica, 75 parts of jujube, 5 parts of ginger, 50 parts of kudzu root, 50 parts of poria, 25 parts of cinnamon bark, and 25 parts of liquorice;

[0551] Extract preparation process: weigh the above medicinal materials, extract, concentrate, and add 25 parts of ginseng extract to the concentrate.

[0552] Fermentation broth preparation process: the extract is fermented by composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or disc centrifuged to obtain the fermentation broth.

[0553] The formula of comparison group 5: 75 parts of angelica, 75 parts of jujube, 75 parts of ginger, 50 parts of polygonatum, 50 parts of kudzu root, 50 parts of poria, 25 parts of cinnamon bark, and 25 parts of liquorice;

[0554] Extract preparation process: weigh the above medicinal materials, extract, concentrate, and add 25 parts of ginseng extract to the concentrate.

[0555] Fermentation broth preparation process: the extract is fermented by composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or disc centrifuged to obtain the fermentation broth.

[0556] The formula of comparison group 6: 75 parts of angelica, 75 parts of jujube, 75 parts of ginger, 50 parts of dried tangerine peel, 50 parts of kudzu root, 50 parts of poria, 25 parts of cinnamon bark, and 25 parts of liquorice;

[0557] Extract preparation process: weigh the above medicinal materials, extract, concentrate, and add 25 parts of ginseng extract to the concentrate.

[0558] Fermentation broth preparation process: the extract is fermented by composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or disc centrifuged to obtain the fermentation broth.

[0559] The formula of comparison group 7: 75 parts of angelica, 75 parts of jujube, 150 parts of ginger, 50 parts of polygonatum, 50 parts of kudzu root, 50 parts of tuckahoe, 50 parts of tangerine peel, 25 parts of cinnamon bark, and 25 parts of liquorice;

[0560] Extract preparation process: weigh the above medicinal materials, extract, concentrate, and add 25 parts of ginseng extract to the concentrate.

[0561] Fermentation broth preparation process: the extract is fermented by composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or disc centrifuged to obtain the fermentation broth.

[0562] The formula of control group 8: 75 parts of angelica, 75 parts of jujube, 75 parts of ginger, 50 parts of kudzu root, 50 parts of poria, 25 parts of cinnamon bark, and 25 parts of liquorice;

[0563] Extract preparation process: weigh the above medicinal materials, extract, concentrate, and add 25 parts of ginseng extract to the concentrate.

[0564] Fermentation broth preparation process: the extract is fermented by composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or disc centrifuged to obtain the fermentation broth.

[0565] Comparative group 9 formula: 50 parts of polygonatum, 50 parts of tangerine peel

[0566] Extract preparation process: weigh the above medicinal materials for extraction and concentration.

[0567] Fermentation broth preparation process: the extract is fermented by composite strains of Lactobacillus plantarum and Lactobacillus rhamnosus, homogenized, filtered or disc centrifuged to obtain the fermentation broth.

[0568] Positive medicine: Bazhenye (batch number: 210710).

[0569] 1.3 Experimental Results

[0570] The results of cell phagocytic index detection are shown in Table 34.

[0571] Table 34 Cell phagocytosis index test results of different formulations

[0572]

[0573] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD, and all experimental group data were analyzed by one-way ANOVA. P < 0.05 was statistically significant. The model group was compared with the blank group ###P < 0.001. The model group, experimental group 2, experimental group 3, the comparison group formula and the positive drug group were compared with the experimental group 1 (the present invention formula), ***P < 0.001; the comparison of the comparison group formula and the positive drug group with the experimental group 2 was ΔΔP < 0.01, ΔΔΔP < 0.001; the comparison of the comparison group formula and the positive drug group with the experimental group 3 was □□P < 0.01, □□□P < 0.001.

[0574] From Table 34, we can see that:

[0575] (1) There was a significant difference between the model group and the blank group (p < 0.001);

[0576] (2) The cell phagocytic index of the experimental group and the control group was better than that of the model group, among which the cell phagocytic index of the experimental group 1 was the highest;

[0577] (3) Compared with experimental group 1, the model group, experimental group 2, experimental group 3, control group formula and positive drug group showed extremely significant differences (p < 0.001), indicating that the ability of experimental group 1 to improve immunity was superior to that of experimental group 2, experimental group 3, control group and positive drug group.

[0578] (4) There were no statistical differences between control groups 6, 7 and 8 and experimental group 2 (p>0.05); there were significant differences between control group 2 and the positive drug group and experimental group 2 (p<0.01); there were extremely significant differences between control group 1, control group 3, control group 4, control group 5 and control group 9 and experimental group 2 (p<0.001).

[0579] (5) There was no statistical difference between the control group 2, control group 6, control group 7, control group 8 and the positive drug group and the experimental group 3 (p>0.05); there was a significant difference between the control group 4 and control group 9 and the experimental group 3 (p<0.01); there was a very significant difference between the control group 1, control group 3 and control group 5 and the experimental group 3 (p<0.001);

[0580] (6) The two formulas of control group 8 and control group 9 are the disassembled formulas of experimental group 1; the cell phagocytosis index of experimental group 1 is increased by 0.267 compared with the model group; the cell phagocytosis index of control group 8 and control group 9 is increased by 0.172 and 0.054 respectively compared with the model group; the calculation results based on Jin's formula show that q = 1.23, q is greater than 1.15, which is a synergistic effect. This shows that the combination of the two formulas of control group 8 and control group 9 has a synergistic enhancement effect, which is better than using them alone.

[0581] Based on the above results, it is shown that the formula of experimental group 1 (the formula of the present invention) obtained by the present invention is the best.

[0582] Example 12 Comparative Experiment 2 on Cell Efficacy of Different Formulas

[0583] 1. Cell SOD activity detection experiment

[0584] 1.1. The experimental cells, consumables, reagents, instruments, methods and steps are the same as those in Example 6.

[0585] 1.2. Different formulations and preparation methods are the same as in Example 11.

[0586] 1.3 Experimental Results

[0587] The results of cell SOD activity detection are shown in Table 35.

[0588] Table 35 Cell SOD activity test results of different formulas (U / mgport)

[0589]

[0590] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD, and all experimental group data were analyzed by one-way ANOVA. P < 0.05 was statistically significant. The model group was compared with the blank group ###P < 0.001. The model group, experimental group 2, experimental group 3, the formula of the comparison group and the positive drug group were compared with the experimental group 1 (the formula of the present invention), ***P < 0.001; the comparison of the formula of the comparison group and the positive drug group with the experimental group 2 was ΔΔP < 0.01; the comparison of the formula of the comparison group and the positive drug group with the experimental group 3 was □P < 0.05, □□P < 0.01, □□□P < 0.001.

[0591] From Table 35, we can see that:

[0592] (1) There was a significant difference between the model group and the blank group (p < 0.001);

[0593] (2) The cellular SOD activities of the experimental and control groups were better than those of the model group, with the SOD in the experimental group 1 being the highest;

[0594] (3) The model group, experimental group 2, experimental group 3, control group and positive drug group showed extremely significant differences compared with experimental group 1 (p < 0.001), indicating that experimental group 1 had the best efficacy.

[0595] (4) There were significant differences between control group 8 and the positive drug group and experimental group 2 (p < 0.01), indicating that their anti-fatigue effects were better than those of experimental group 2. There were significant differences between control group 3 and experimental group 2 (p < 0.01); there was no statistical difference between control group 6 and experimental group 2 (p > 0.05); there was a significant difference between control group 9 and experimental group 2 (p < 0.01); indicating that the efficacy of experimental group 2 was no different from control group 6 but was better than control group 3 and control group 9.

[0596] (5) There were significant differences between the control groups 3 and 8 and the experimental group 3 (p < 0.01); there was no statistical difference between the control group 6 and the experimental group 3 (p > 0.05); there was a very significant difference between the control group 9 and the experimental group 3 (p < 0.001), and there was a very significant difference between the positive drug group and the experimental group 3 (p < 0.001), indicating that the efficacy of the experimental group 3 was better than that of the control groups 3 and 9; the efficacy of the control group 8 and the positive drug group was better than that of the experimental group 3. There was no difference in efficacy between the control group 6 and the experimental group 3.

[0597] (6) The two formulas of control group 8 and control group 9 are the disassembled formulas of experimental group 1; the cell SOD activity of experimental group 1 is increased by 4.018U / mgport compared with the model group; the cell phagocytic index of control group 8 and control group 9 is increased by 2.611U / mgport and 0.591U / mgport respectively compared with the model group; the calculation results based on King's formula show that q=2.42, q is greater than 1.15, which is a synergistic effect. This shows that the combination of the two formulas of control group 8 and control group 9 has a synergistic enhancement effect, which is better than using them alone.

[0598] Based on the above results, it is shown that the formula of experimental group 1 (the formula of the present invention) obtained by the present invention is the best.

[0599] 2. Cellular reduced glutathione (GSH) detection experiment

[0600] 2.1. Different formulations and preparation methods are the same as in Example 11.

[0601] 2.2. Raw264.7 Cell GSH Experimental Methods and Steps

[0602] (1) Raw264.7 cell culture

[0603] RAW264.7 cells were inoculated in DMEM medium (containing 10% fetal bovine serum, 100 U / mL penicillin, 100 U / mL streptomycin) and cultured in a 37°C, 5% CO2 incubator until the cells adhered to the wall and grew to 70% to 80% (i.e., subculture, generally 24 hours).

[0604] (2) Establishment of H2O2-induced oxidative stress model in RAW264.7 cells

[0605] RAW264.7 cells in the logarithmic growth phase were taken and diluted to 1×10 using DMEM complete culture medium containing 10% fetal bovine serum. 6 cells / mL; 2 mL of cell suspension was added to each 10 cm culture dish and cultured in a cell culture incubator at 37°C and 5% CO2 for 24 h.

[0606] Discard the supernatant, rinse twice with PBS, add 8 mL of 1000 μM H2O2 solution diluted with DMEM medium to the plate wells, and culture for 24 hours. Blank control group: add cells to culture for 24 hours, discard the supernatant and add 8 mL of serum-free DMEM medium. After the Raw 264.7 cell oxidative stress model is established, discard the original culture medium, dilute the samples into groups, and add them to the culture dish for 24 hours.

[0607] (3) Perform the test according to the steps of the reduced glutathione (GSH) content detection kit (Solebo Biochemical Kit Division, product number BC1175).

[0608] 2.3 Experimental Results

[0609] The results of cellular GSH detection are shown in Table 36.

[0610] Table 36 Cellular GSH detection results of different formulas (ug / mgprot)

[0611]

[0612] Note: GraphpadPrism 9.5.1 software was used for statistical analysis of data. All experimental data were expressed as mean + SD, and all experimental group data were analyzed by one-way ANOVA. P < 0.05 was statistically significant. The model group was compared with the blank group ###P < 0.001. The model group, experimental group 2, experimental group 3, the comparison group formula and the positive drug group were compared with the experimental group 1 (the present invention formula), ***P < 0.001; the comparison of the comparison group formula and the positive drug group with the experimental group 2 was ΔP < 0.05, ΔΔΔP < 0.001; the comparison of the comparison group formula and the positive drug group with the experimental group 3 was □□P < 0.01, □□□P < 0.001.

[0613] From Table 36, we can see that:

[0614] (1) There was a significant difference between the model group and the blank group (p < 0.001);

[0615] (2) The cellular GSH of both the experimental and control groups was higher than that of the model group, among which the GSH of experimental group 1 was the highest;

[0616] (3) Compared with experimental group 1, the model group, experimental group 2, experimental group 3, control group and positive drug group showed extremely significant differences (p < 0.001), indicating that the antioxidant effect of experimental group 1 was better than that of experimental group 2, experimental group 3, control groups 1 to 9 and positive drug group.

[0617] (4) The GSH test results of the positive drug group were better than those of the experimental group 2, and there was a very significant difference compared with the experimental group 2 (p < 0.001), indicating that its anti-fatigue effect was better than that of the experimental group 2. There was no statistical difference between the control group 8 and the experimental group 2 (p > 0.05). There was a statistical difference between the control group 1 and the experimental group 2 (p < 0.05); the control groups 2, 3, 5, 6 and 9 had a very significant difference compared with the experimental group 2 (p < 0.001), indicating that the antioxidant effect of the experimental group 2 was the best.

[0618] (5) The GSH test results of the positive drug group were better than those of the experimental group 3, and there was a very significant difference compared with the experimental group 3 (p < 0.001), indicating that its anti-fatigue effect was better than that of the experimental group 3; there was no statistical difference between the control group 8 and the experimental group 3 (p > 0.05); the control groups 1, 2, 3, 5, 6 and 9 were very significantly different from the experimental group 3 (p < 0.001), indicating that the antioxidant effect of the experimental group 3 was the best.

[0619] Based on the above results, it is shown that the formula of experimental group 1 (the formula of the present invention) obtained by the present invention is the best.

[0620] Example 13: Human body test verification of ginseng and polygonatum fermented plant beverage

[0621] 1.1 Test Service Personnel

[0622] A total of 9 people with insufficient Qi and blood were selected to volunteer for the human trial.

[0623] 1.2 Trial Recipe

[0624] 1.2.1 Formula of compound fermented beverage of traditional Chinese medicine composition

[0625] 75 parts of Chinese Angelica, 75 parts of Chinese dates, 75 parts of ginger, 50 parts of polygonatum, 50 parts of kudzu root, 50 parts of tuckahoe, 50 parts of dried orange peel, 25 parts of cinnamon, 25 parts of liquorice, 25 parts of ginseng extract, 350 parts of oligofructose, 100 parts of xylitol, 200 parts of erythritol, 100 parts of black wolfberry juice powder, 5 parts of taurine, 10 parts of gamma-aminobutyric acid

[0626] 1.2.2 Preparation method of compound fermented beverage of Chinese medicine composition

[0627] The preparation method of the Chinese medicine composition fermented beverage is the same as that of Example 4

[0628] 1.3 Trial method and results, see Table 37.

[0629] Table 37 Human test methods and results

[0630]

[0631]

[0632] 1.3 Conclusion

[0633] According to the above trial results, it can be concluded that after people with insufficient qi and blood try the product of the present invention, they feel lighter and in a better mental state; after people who are prone to fatigue for a long time take this product, their mental state improves and fatigue is significantly relieved. This shows that this product can improve the mental state of people with chronic fatigue and significantly relieve fatigue by nourishing qi and blood.

[0634] Example 14 Clinical Trial

[0635] A randomized double-blind placebo-controlled clinical trial of 240 cases for the prevention and treatment of chronic fatigue syndrome was conducted. The results of the clinical trial showed that the patients taking the product of the present invention had significant improvements in various symptoms of chronic fatigue such as qi and blood deficiency and fatigue.

[0636] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A Chinese medicinal fermentation composition that is both medicinal and edible, characterized in that: Including ginseng, angelica, jujube, ginger, polygonatum, kudzu root, poria, tangerine peel, cinnamon and licorice; The bacteria used in the fermentation include probiotics.

2. The Chinese medicinal fermentation composition as claimed in claim 1, characterized in that: The edible and medicinal Chinese medicine fermentation composition comprises, by weight: Preferably, the Chinese medicinal fermentation composition for food and medicine comprises, by weight: 50-130 parts of ginseng, 50-130 parts of angelica, 50-130 parts of jujube, 50-130 parts of ginger, 30-90 parts of polygonatum, 30-90 parts of kudzu root, 30-90 parts of tuckahoe, 30-90 parts of tangerine peel, 20-40 parts of cinnamon and 20-40 parts of liquorice; or More preferably, the edible and medicinal Chinese medicine fermentation composition comprises, by weight: 60-110 parts of ginseng, 60-110 parts of angelica, 60-110 parts of jujube, 60-110 parts of ginger, 40-80 parts of polygonatum, 40-80 parts of kudzu root, 40-80 parts of tuckahoe, 40-80 parts of tangerine peel, 20-30 parts of cinnamon and 20-30 parts of liquorice; or More preferably, the edible and medicinal Chinese medicine fermentation composition comprises, by weight: 60-90 parts of ginseng, 60-90 parts of angelica, 60-90 parts of jujube, 60-90 parts of ginger, 40-60 parts of polygonatum, 40-60 parts of kudzu root, 40-60 parts of poria, 40-60 parts of tangerine peel, 20-30 parts of cinnamon and 20-30 parts of liquorice.

3. The Chinese medicinal fermentation composition as claimed in claim 1 or 2, characterized in that: The edible and medicinal Chinese medicine fermentation composition comprises, by weight: 75 parts of ginseng, 75 parts of angelica, 75 parts of jujube, 75 parts of ginger, 50 parts of polygonatum, 50 parts of kudzu root, 50 parts of tuckahoe, 50 parts of tangerine peel, 25 parts of cinnamon and 25 parts of liquorice; or 130 parts of ginseng, 50 parts of angelica, 130 parts of jujube, 50 parts of ginger, 90 parts of polygonatum, 40 parts of kudzu root, 40 parts of tuckahoe, 60 parts of tangerine peel, 30 parts of cinnamon and 20 parts of liquorice; or 90 parts of ginseng, 90 parts of angelica, 60 parts of jujube, 60 parts of ginger, 40 parts of polygonatum, 60 parts of kudzu root, 60 parts of tuckahoe, 40 parts of tangerine peel, 20 parts of cinnamon and 30 parts of liquorice; or 75 parts of ginseng, 75 parts of angelica, 75 parts of jujube, 30 parts of ginger, 50 parts of polygonatum, 50 parts of kudzu root, 50 parts of tuckahoe, 50 parts of tangerine peel, 25 parts of cinnamon and 25 parts of liquorice; or 75 parts of the ginseng, 75 parts of the angelica, 75 parts of the jujube, 140 parts of the ginger, 50 parts of the polygonatum, 50 parts of the kudzu root, 50 parts of the tuckahoe, 50 parts of the tangerine peel, 25 parts of the cinnamon and 25 parts of the liquorice.

4. The Chinese medicinal fermentation composition as claimed in any one of claims 1 to 3, characterized in that: The ginseng in the Chinese medicinal fermentation composition for food and medicine includes ginseng extract and / or ginseng concentrate; or More preferably, the ginseng extract or the ginseng concentrate is obtained by extracting ginseng and water in a ratio of 3 to 3.5:1 (W / W); or More preferably, the content of total ginsenosides in the ginseng extract or the ginseng concentrate is ≥3%.

5. The Chinese medicinal fermentation composition as claimed in claim 1, characterized in that: The probiotics include one or more of Lactobacillus, Lactobacillus, Lactobacillus and / or Lactobacillus mucilaginosus; or More preferably, the genus Lactobacillus includes one or more of Lactobacillus casei, Lactobacillus paracasei and / or Lactobacillus rhamnosus; the genus Lactobacillus includes Lactobacillus plantarum; the genus Lactobacillus includes one or more of Lactobacillus delbrueckii subspecies bulgaricus and / or Lactobacillus helveticus; the genus Lactobacillus mucosus includes Lactobacillus fermentum; or More preferably, the probiotics include the Lactobacillus genus and the Lactobacillus genus; or More preferably, the probiotics include Lactobacillus plantarum and Lactobacillus rhamnosus; or More preferably, the inoculation amount of the probiotics comprises 2-10% (V / V); 或 Preferably, the inoculation amount of the probiotics comprises 2%, 3%, 4%, 5% or 6% (V / V); or more preferably, the inoculation amount of the probiotics comprises 4% (V / V); Or the inoculation ratio of the Lactobacillus genus and the Lactobacillus genus in the probiotics includes (1-3): (1-3); or preferably, the inoculation ratio of the plant lactobacillus and the rhamnosus lactobacillus in the probiotics is 1:1; the OD of the plant lactobacillus in the probiotics is 600 Including 3.0 to 5.0, the OD of the rhamnosus Lactobacillus 600 Including 3.5~5.5; The pH of the probiotic inoculation includes 6.5; Or the fermentation time of the probiotic fermentation includes 16 to 48 hours; Or preferably, the fermentation time of the probiotic fermentation includes 20 hours, 24 hours, 26 hours, 28 hours or 32 hours; or preferably, the fermentation time of the probiotic fermentation includes 24 to 32 hours.

6. The method for preparing the edible and medicinal Chinese medicine fermentation composition according to any one of claims 1 to 5, characterized in that: Weigh the raw materials according to the ratio, extract, concentrate, ferment, mix and fill to obtain the Chinese medicinal fermentation composition of food and medicine; The extraction includes but is not limited to 1 extraction or 2 extractions; Or preferably, the extraction includes one extraction; preferably, the amount of water added in the one extraction includes 8 to 12 times of the raw material; more preferably, the amount of water added in the one extraction includes 10 times of the raw material; preferably, the time for one extraction is 1 to 3 hours; more preferably, the time for one extraction is 1.5 hours; Or preferably, the extraction comprises 2 extractions; The amount of water added in the two extractions includes 14 to 22 times of the raw material, the amount of water added in the first of the two extractions includes 8 to 12 times of the raw material, and the amount of water added in the second of the two extractions includes 6 to 10 times of the raw material; Preferably, the amount of water added in the first extraction of the two extractions includes 10 or 12 times the amount of the raw material, and the amount of water added in the second extraction of the two extractions includes 8 times the amount of the raw material; More preferably, the amount of water added in the first extraction of the two extractions includes 10 times the amount of the raw material; The time for the two extractions includes 1 to 3 hours, the time for the first extraction of the two extractions includes 1 to 2 hours, and the time for the second extraction of the two extractions includes 1 to 2 hours; Preferably, the time for the first extraction of the two extractions includes 1 hour, 1.5 hours or 2 hours, and the time for the second extraction of the two extractions includes 1 hour; The concentration includes but is not limited to double-effect concentration; the multiple of concentration includes but is not limited to 3 times; The fermentation temperature includes but is not limited to 37° C., the rotation speed includes but is not limited to 50 rpm, the tank pressure includes 0.04-0.06 MPa; and the pH at the end of the fermentation includes 3.3-3.

8.

7. The Chinese medicinal fermentation composition with both medicinal and edible properties obtained by the preparation method as claimed in claim 6.

8. A compound fermented product of a Chinese medicine composition having the same medicinal and edible properties, characterized in that: The fermented Chinese medicinal composition comprising the medicinal and edible fermented composition as claimed in any one of claims 1 to 5 or the medicinal and edible fermented Chinese medicinal composition as claimed in claim 7; Preferably, the compound fermented product of the Chinese medicine composition with the same origin as medicine and food further comprises one or more of oligofructose, γ-aminobutyric acid and / or taurine; the oligofructose comprises oligofructose powder and / or oligofructose liquid; Preferably, the compound fermented product of the Chinese medicine composition with both medicinal and edible properties further comprises one or more of erythritol, xylitol and / or black wolfberry juice powder.

9. The Chinese medicine composition compound fermented product according to claim 8, characterized in that: In parts by weight, the compound fermented product of the Chinese medicine composition with the same medicinal and edible properties also includes: 100-700 parts of the oligofructose, 5-20 parts of the γ-aminobutyric acid and / or 2-10 parts of the taurine; Preferably, the compound fermented product of the Chinese medicine composition with the same origin as food and medicine further comprises, by weight: 200-500 parts of the oligofructose, 5-10 parts of the γ-aminobutyric acid and / or 2-5 parts of the taurine; More preferably, the compound fermented product of the Chinese medicine composition with the same medicinal and edible properties further comprises, by weight: 100-300 parts of the oligofructose, 5-10 parts of the γ-aminobutyric acid and / or 2-5 parts of the taurine; More preferably, the compound fermented product of the Chinese medicine composition with the same medicinal and edible properties further comprises, by weight: 350 parts of the oligofructose, 10 parts of the γ-aminobutyric acid and / or 5 parts of the taurine; or 250 parts of the oligofructose, 10 parts of the γ-aminobutyric acid and / or 5 parts of the taurine; or 200 parts of the oligofructose, 5 parts of the gamma-aminobutyric acid and / or 2 parts of the taurine; or 300 parts of the oligofructose, 20 parts of the gamma-aminobutyric acid and / or 5 parts of the taurine; Preferably, the compound fermented product of the Chinese medicine composition with the same origin as food and medicine further comprises, by weight: 100-400 parts of erythritol, 50-200 parts of xylitol and / or 50-200 parts of black wolfberry juice powder; Preferably, the compound fermented product of the Chinese medicine composition with the same origin as food and medicine further comprises, by weight: 150-300 parts of erythritol, 80-150 parts of xylitol and / or 80-150 parts of black wolfberry juice powder; More preferably, the compound fermented product of the Chinese medicine composition with the same origin as food and medicine further comprises, by weight: 200 parts of the erythritol, 100 parts of the xylitol and / or 100 parts of the black wolfberry juice powder; or 100 parts of the erythritol, 150 parts of the xylitol and / or 150 parts of the black wolfberry juice powder; or 300 parts of the erythritol, 80 parts of the xylitol and / or 80 parts of the black wolfberry juice powder.

10. Use of any of the following in the preparation of nourishing, anti-fatigue, immunity-enhancing and / or anti-oxidative products: (I) A Chinese medicinal fermentation composition as claimed in any one of claims 1 to 5; (II), the Chinese medicinal fermentation composition of claim 7; or (III) The compound fermented product of the Chinese medicine composition with both medicinal and edible properties as described in claim 8 or 9.

11. The use according to claim 10, characterized in that The anti-fatigue includes anti-chronic fatigue syndrome; The anti-oxidation includes increasing the total antioxidant activity and / or increasing the cellular reduced glutathione content; The improving of the total antioxidant activity comprises improving one or more of the following: the activity of superoxide dismutase in cells, the scavenging rate of DPPH free radicals and / or the scavenging rate of hydroxyl free radicals; The enhancement of immunity includes but is not limited to increasing the cell phagocytic index.

12. A product, characterized in that Include any of the following and acceptable excipients; (I) A Chinese medicinal fermentation composition as claimed in any one of claims 1 to 5; (II), the Chinese medicinal fermentation composition of claim 7; or (III) The compound fermented product of the Chinese medicine composition with both medicinal and edible properties as described in claim 8 or 9.

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