A plant fermentation product with soothing and repairing effects, and its preparation method and application

Abstract

The present invention belongs to the field of microbial fermentation technology, and discloses a plant fermentation product with soothing and repairing effects, as well as its preparation method and application. The raw materials for preparing the plant fermentation product include: plant raw materials, solvents and fermentation strains; the plant raw materials include sea buckthorn fruit and licorice; the solvents include solvent A and solvent B, solvent A includes glycerol and / or ethoxydiglycol, and solvent B includes water; the fermentation strains include at least one of Lactobacillus plantarum, Lactobacillus casei and Lactobacillus fermentum. The present invention organically combines high-pressure homogenization and specific microbial fermentation, and high-pressure homogenization is supplemented with a specific solvent, so that plant tissue cells can be fully broken, components can be fully released and dissolved, and the utilization efficiency of plant raw materials can be improved. The plant composite liquid obtained by high-pressure homogenization is directly used as the subsequent fermentation matrix. Through the biotransformation ability of microorganisms, a plant fermentation product with a high content of active substances and significant soothing and repairing effects can be prepared.

Description

Technical Field

[0001] The present invention belongs to the technical field of microbial fermentation, and in particular relates to a plant fermentation product with soothing and repairing effects, and a preparation method and application thereof. Background Art

[0002] Licorice is the root and rhizome of the common plant of the genus Glycyrrhiza in the legume family. The "Chinese Pharmacopoeia" (2020 edition) stipulates that its plant sources include licorice (Glycyrrhiza uralensis Fisch.), licorice (Glycyrrhiza inflata Bat.) and licorice (Glycyrrhiza glabra L.). Licorice has been a very important medicinal plant since ancient times. It has the effects of tonifying the spleen and replenishing qi, clearing away heat and detoxifying, removing phlegm and relieving cough, relieving acute pain, and harmonizing various medicines. It is widely used in the field of medicine and health. At the same time, the above three types of licorice are included in the "Catalogue of Used Cosmetic Raw Materials" (2021 edition). They have excellent whitening, anti-inflammatory, soothing, repairing and antioxidant effects, and are widely used in the cosmetics industry as active raw materials. The main active ingredients of licorice include flavonoids and terpenes. Among them, the flavonoid components of licorice that have been studied in depth include liquiritin, isoliquiritin, liquiritigenin and glabridin. The representative terpenoid components in licorice are glycyrrhizic acid. They are all ingredients with poor water solubility and are currently mainly extracted using organic solvents such as ethanol. The production and preparation of these active ingredients puts tremendous pressure on the environment and safety.

[0003] Seabuckthorn (Hippophae rhamnoides L.), a edible and medicinal plant of the genus Hippophae in the family Elaeagnaceae, possesses excellent antioxidant, anti-ulcer, antibacterial, and anticancer properties. Most of its active ingredients are found in its fruit, including flavonoids, fatty acids, oils, and vitamins. Seabuckthorn flavonoids primarily include isorhamnetin, quercetin, kaempferol, glycosides, and rutin. Flavonoids, fatty acids, and oils are all poorly water-soluble active ingredients.

[0004] Plant extracts obtained by traditional water extraction and alcohol extraction techniques cannot extract the active ingredients from plant raw materials in a green and efficient manner. The active ingredients contained in the extracts are limited and cannot fully exert the efficacy of the plant raw materials.

[0005] Therefore, how to increase the active ingredients in plant extracts is a technical problem that needs to be solved urgently in this field. Summary of the Invention

[0006] The present invention aims to solve at least one of the technical problems existing in the above-mentioned prior art. To this end, the present invention provides a plant fermentation product with soothing and repairing effects, as well as a preparation method and application thereof. The plant fermentation product has a high content of flavonoids and triterpenoid active ingredients, thereby having significant soothing and repairing effects.

[0007] A first aspect of the present invention provides a plant fermentation product, wherein the raw materials for preparing the plant fermentation product include: a plant raw material, a solvent and a fermentation strain; the plant raw material includes sea buckthorn fruit and licorice; the solvent includes solvent A and solvent B, the solvent A includes glycerol and / or ethoxydiglycol, and the solvent B includes water; the fermentation strain includes at least one of Lactobacillus plantarum, Lactobacillus casei and Lactobacillus fermentum.

[0008] According to some embodiments of the present invention, the ratio of the seabuckthorn fruit, the licorice root, and the solvent is (0.001-0.1g):(0.001-0.1g):1g. Further, the ratio of the seabuckthorn fruit, the licorice root, and the solvent is (0.01-0.1g):(0.01-0.1g):1g.

[0009] According to some embodiments of the present invention, the ratio of the amount of solvent A to the amount of solvent B is (0.01-5g):1g. Further, the ratio of the amount of solvent A to the amount of solvent B is (0.01-1g):1g. Further, the ratio of the amount of solvent A to the amount of solvent B is (0.1-1g):1g.

[0010] According to some embodiments of the invention, the licorice comprises at least one of Glycyrrhiza glabra, Glycyrrhiza inflata and Glycyrrhiza uralensis.

[0011] According to some embodiments of the present invention, the concentration of the fermentation strain is 10 5 -10 10 CFU / mL. Further, the concentration of the fermentation strain is 10 5 -10 8 CFU / mL. Further, the concentration of the fermentation strain is 10 8 -10 10 CFU / mL.

[0012] The second aspect of the present invention provides a method for preparing the plant fermentation product according to the first aspect of the present invention, comprising the following steps:

[0013] Mixing the plant raw material and the solvent, and homogenizing under high pressure to obtain a plant composite liquid;

[0014] The plant complex liquid and the fermentation strain are mixed and fermented to obtain the plant fermentation product.

[0015] According to some embodiments of the present invention, before the fermentation, the process further includes adding a culture medium; the ratio of the plant complex solution, the fermentation strain, and the culture medium is (1-500 g): (0.01-10 mL): 1 g. Furthermore, the ratio of the plant complex solution, the fermentation strain, and the culture medium is (1-250 g): (0.1-10 mL): 1 g. Furthermore, the ratio of the plant complex solution, the fermentation strain, and the culture medium is (10-100 g): (1-10 mL): 1 g.

[0016] According to some embodiments of the present invention, the culture medium is MRS medium.

[0017] According to some embodiments of the present invention, the pressure of the high-pressure homogenization is 10-100 MPa; and / or the temperature of the high-pressure homogenization is 25-50°C; and / or the time of the high-pressure homogenization is 5-40 minutes. Further, the pressure of the high-pressure homogenization is 40-100 MPa. Further, the pressure of the high-pressure homogenization is 60-100 MPa. Further, the temperature of the high-pressure homogenization is 30-50°C. Further, the temperature of the high-pressure homogenization is 35-45°C. Further, the time of the high-pressure homogenization is 5-30 minutes. Further, the time of the high-pressure homogenization is 10-20 minutes.

[0018] According to some embodiments of the present invention, the fermentation temperature is 20-45°C; and / or the fermentation time is 12-120 hours. Further, the fermentation temperature is 20-40°C. Further, the fermentation temperature is 22-35°C. Further, the fermentation time is 30-110 hours. Further, the fermentation time is 50-100 hours.

[0019] According to some embodiments of the present invention, after the fermentation, the method further comprises sterilizing the fermentation broth.

[0020] According to some embodiments of the present invention, the sterilization temperature is 100-150° C., and the sterilization time is 10-30 min.

[0021] According to some embodiments of the present invention, the method further comprises centrifuging the sterilized fermentation broth.

[0022] According to some embodiments of the present invention, the rotation speed of the centrifugal treatment is 5000-7000 rpm / min, and the time of the centrifugal treatment is 10-30 min.

[0023] The third aspect of the present invention provides a cosmetic comprising the plant fermentation product according to the first aspect of the present invention.

[0024] According to some embodiments of the present invention, the cosmetic further comprises an auxiliary material acceptable in cosmetics.

[0025] According to some embodiments of the present invention, the acceptable excipients in the cosmetic include at least one of an emulsifier, a moisturizer, a whitening agent, and a preservative.

[0026] According to some embodiments of the present invention, the amount of the plant fermentation product added to the cosmetic is 0.01-80 wt %. Further, the amount of the plant fermentation product added to the cosmetic is 1-50 wt %. Further, the amount of the plant fermentation product added to the cosmetic is 1-10 wt %.

[0027] According to some embodiments of the present invention, the cosmetic is an emulsion, and the components of the emulsion include the plant fermentation product described in the first aspect of the present invention, octyldodecanol, undecylenoyl phenylalanine, PEG-40 sodium stearate, 1,2-hexanediol, p-hydroxyacetophenone and water.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] The present invention uses a specific solvent in the high-pressure homogenization process to fully break up the tissue cells of the plant raw material; at the same time, the oil in the sea buckthorn fruit is used to form a microemulsion during the high-pressure homogenization process, thereby improving the dissolution rate of active ingredients of different polarities. It is also organically combined with the directional microbial fermentation technology and fermented using a specific fermentation strain. The plant composite liquid obtained by high-pressure homogenization is used as the subsequent fermentation matrix. Through the biotransformation ability of the fermentation strain, the content of active ingredients in the plant fermentation product is ultimately increased, including total flavonoids, four flavonoid components (isorhamnetin, liquiritin, licorice chalcone A, and glycyrrhizin) and one triterpenoid (glycyrrhizic acid), so that the plant fermentation product of the present invention has significant soothing and repairing effects.

[0030] This invention utilizes plant active ingredient extraction technology developed through high-pressure homogenization combined with modern microbial fermentation engineering to develop unique plant-microbial raw materials. Microorganisms produce numerous bioactive ingredients during their growth, including enzymes, polysaccharides, peptides, and small molecule compounds. Some of these ingredients have a potent volume-enhancing effect and can increase the solubility of poorly soluble ingredients in the fermentation system, thereby increasing the content of some active ingredients and resulting in a more effective skin-protecting product overall. DETAILED DESCRIPTION

[0031] In order to make the technical solution of the present invention more clearly understood by those skilled in the art, the following examples are given for illustration. It should be noted that the following examples do not limit the scope of protection claimed by the present invention.

[0032] Unless otherwise specified, the raw materials, reagents, or devices used in the following examples can be obtained from conventional commercial sources or by existing known methods.

[0033] Example 1

[0034] A plant fermentation product, the preparation raw materials of which include: sea buckthorn fruit, glycyrrhiza glabra, ethoxydiglycol, deionized water, lactobacillus plantarum liquid and MRS culture medium.

[0035] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0036] 1) 20 g of sea buckthorn fruit, 20 g of glycyrrhiza glabra, 200 g of ethoxydiglycol, and 400 g of deionized water were mixed, and then added to a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0037] 2) 300g plant compound liquid, 6mL plant lactobacillus liquid (concentration of 10 8 CFU / mL) and 4.5 g MRS medium were mixed and fermented at 28°C for 72 h. After the fermentation, the fermentation broth was sterilized at 121°C for 20 min, and the sterilized fermentation broth was centrifuged at 6000 rpm / min for 20 min. The supernatant after centrifugation was obtained to obtain a plant fermentation product.

[0038] Example 2

[0039] A plant fermentation product, the preparation raw materials of which include: sea buckthorn fruit, glycyrrhiza inflata, ethoxydiglycol, deionized water, lactobacillus plantarum liquid and MRS culture medium.

[0040] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0041] 1) 20 g of sea buckthorn fruit, 20 g of Glycyrrhiza inflata, 200 g of ethoxydiglycol, and 400 g of deionized water were mixed, and then added to a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0042] 2) 300g plant compound liquid, 6mL plant lactobacillus liquid (concentration of 10 8 CFU / mL) and 4.5 g MRS medium were mixed and fermented at 28°C for 72 h. After the fermentation, the fermentation broth was sterilized at 121°C for 20 min, and the sterilized fermentation broth was centrifuged at 6000 rpm / min for 20 min. The supernatant after centrifugation was obtained to obtain a plant fermentation product.

[0043] Example 3

[0044] A plant fermentation product, the preparation raw materials of which include: sea buckthorn fruit, licorice, ethoxydiglycol, deionized water, lactobacillus plantarum liquid and MRS culture medium.

[0045] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0046] 1) 20 g of sea buckthorn fruit, 20 g of Ural liquorice, 200 g of ethoxydiglycol, and 400 g of deionized water were mixed, and then added to a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0047] 2) 300g plant compound liquid, 6mL plant lactobacillus liquid (concentration of 10 8 CFU / mL) and 4.5 g MRS medium were mixed and fermented at 28°C for 72 h. After the fermentation, the fermentation broth was sterilized at 121°C for 20 min, and the sterilized fermentation broth was centrifuged at 6000 rpm / min for 20 min. The supernatant after centrifugation was obtained to obtain a plant fermentation product.

[0048] Example 4

[0049] A plant fermentation product, the preparation raw materials of which include: sea buckthorn fruit, glycyrrhiza glabra, glycerin, deionized water, lactobacillus plantarum liquid and MRS culture medium.

[0050] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0051] 1) 30 g of sea buckthorn fruit, 10 g of licorice root, 250 g of glycerin, and 350 g of deionized water were mixed, and then added to a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0052] 2) 300g plant compound liquid, 6mL plant lactobacillus liquid (concentration of 10 8 CFU / mL) and 4.5 g MRS medium were mixed and fermented at 28°C for 72 h. After the fermentation, the fermentation broth was sterilized at 121°C for 20 min, and the sterilized fermentation broth was centrifuged at 6000 rpm / min for 20 min. The supernatant after centrifugation was obtained to obtain a plant fermentation product.

[0053] Example 5

[0054] A plant fermentation product, the preparation raw materials of which include: sea buckthorn fruit, glycyrrhiza inflata, glycerol, deionized water, lactobacillus plantarum liquid and MRS culture medium.

[0055] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0056] 1) 30 g of seabuckthorn fruit, 10 g of Glycyrrhiza inflata, 250 g of glycerin, and 350 g of deionized water were mixed, and then added to a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0057] 2) 300g plant compound liquid, 6mL plant lactobacillus liquid (concentration of 10 8 CFU / mL) and 4.5 g MRS medium were mixed and fermented at 28°C for 72 h. After the fermentation, the fermentation broth was sterilized at 121°C for 20 min, and the sterilized fermentation broth was centrifuged at 6000 rpm / min for 20 min. The supernatant after centrifugation was obtained to obtain a plant fermentation product.

[0058] Example 6

[0059] A plant fermentation product, the preparation raw materials of which include: sea buckthorn fruit, ural licorice, glycerol, deionized water, plant lactobacillus liquid and MRS culture medium.

[0060] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0061] 1) 30 g of seabuckthorn fruit, 10 g of Ural liquorice, 250 g of glycerin, and 350 g of deionized water were mixed, and then added to a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0062] 2) 300g plant compound liquid, 6mL plant lactobacillus liquid (concentration of 10 8 CFU / mL) and 4.5 g MRS medium were mixed and fermented at 28°C for 72 h. After the fermentation, the fermentation broth was sterilized at 121°C for 20 min, and the sterilized fermentation broth was centrifuged at 6000 rpm / min for 20 min. The supernatant after centrifugation was obtained to obtain a plant fermentation product.

[0063] Example 7

[0064] A plant fermentation product, the preparation raw materials of which include: sea buckthorn fruit, glycyrrhiza inflata, ethoxydiglycol, glycerol, deionized water, lactobacillus plantarum liquid and MRS culture medium.

[0065] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0066] 1) 20 g of sea buckthorn fruit, 20 g of Glycyrrhiza inflata, 100 g of ethoxydiglycol, 150 g of glycerin, and 350 g of deionized water were mixed, and then added to a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0067] 2) 300g plant compound liquid, 6mL plant lactobacillus liquid (concentration of 10 8 CFU / mL) and 4.5 g MRS medium were mixed and fermented at 28°C for 72 h. After the fermentation, the fermentation broth was sterilized at 121°C for 20 min, and the sterilized fermentation broth was centrifuged at 6000 rpm / min for 20 min. The supernatant after centrifugation was obtained to obtain a plant fermentation product.

[0068] Example 8

[0069] A plant fermentation product, the preparation raw materials of which include: sea buckthorn fruit, glycyrrhiza glabra, glycyrrhiza inflata, glycyrrhiza uralensis, ethoxydiglycol, glycerol, deionized water, lactobacillus plantarum bacterial liquid and MRS culture medium.

[0070] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0071] 1) 10 g of seabuckthorn fruit, 10 g of glycyrrhiza glabra, 10 g of glycyrrhiza inflata, 10 g of uralensis, 150 g of ethoxydiglycol, 150 g of glycerin, and 300 g of deionized water were mixed, and then added to a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0072] 2) 300g plant compound liquid, 6mL plant lactobacillus liquid (concentration of 10 8 CFU / mL) and 4.5 g MRS medium were mixed and fermented at 28°C for 72 h. After the fermentation, the fermentation broth was sterilized at 121°C for 20 min, and the sterilized fermentation broth was centrifuged at 6000 rpm / min for 20 min. The supernatant after centrifugation was obtained to obtain a plant fermentation product.

[0073] Example 9

[0074] Compared with Example 1, the only difference in this embodiment is that the Lactobacillus plantarum liquid is replaced with Lactobacillus casei liquid, the amount of the liquid added is the same as that of the Lactobacillus plantarum liquid in Example 1, and the other components, addition amounts and preparation methods are the same as those in Example 1.

[0075] Example 10

[0076] Compared with Example 2, the only difference in this embodiment is that the Lactobacillus plantarum bacterial liquid is replaced with Lactobacillus casei bacterial liquid, the amount of bacterial liquid added is the same as that of the Lactobacillus plantarum bacterial liquid in Example 2, and the other components, addition amounts and preparation method are the same as those in Example 2.

[0077] Example 11

[0078] Compared with Example 3, the only difference in this embodiment is that the Lactobacillus plantarum bacterial solution is replaced with Lactobacillus casei bacterial solution, the amount of bacterial solution added is the same as that of the Lactobacillus plantarum bacterial solution in Example 3, and the other components, addition amounts and preparation method are the same as those in Example 3.

[0079] Example 12

[0080] Compared with Example 4, the only difference between this embodiment is that the Lactobacillus plantarum bacterial solution is replaced with Lactobacillus casei bacterial solution, the amount of bacterial solution added is the same as that of the Lactobacillus plantarum bacterial solution in Example 4, and the other components, addition amounts and preparation methods are the same as those in Example 4.

[0081] Example 13

[0082] Compared with Example 5, the only difference between this embodiment is that the Lactobacillus plantarum bacterial solution is replaced with Lactobacillus casei bacterial solution, the amount of bacterial solution added is the same as that of the Lactobacillus plantarum bacterial solution in Example 5, and the other components, addition amounts and preparation method are the same as those in Example 5.

[0083] Example 14

[0084] Compared with Example 6, the only difference in this embodiment is that the Lactobacillus plantarum bacterial solution is replaced with Lactobacillus casei bacterial solution, the amount of bacterial solution added is the same as that of the Lactobacillus plantarum bacterial solution in Example 6, and the other components, addition amounts and preparation methods are the same as those in Example 6.

[0085] Example 15

[0086] Compared with Example 7, the only difference between this embodiment is that the Lactobacillus plantarum liquid is replaced with Lactobacillus casei liquid, the amount of the liquid added is the same as that of the Lactobacillus plantarum liquid in Example 7, and the other components, addition amounts and preparation method are the same as those in Example 7.

[0087] Example 16

[0088] Compared with Example 8, the only difference between this embodiment is that the Lactobacillus plantarum liquid is replaced with Lactobacillus casei liquid, the amount of the liquid added is the same as that of the Lactobacillus plantarum liquid in Example 8, and the other components, addition amounts and preparation method are the same as those in Example 8.

[0089] Example 17

[0090] Compared with Example 1, the only difference between this embodiment is that the Lactobacillus plantarum liquid is replaced with Lactobacillus fermentum liquid, and the amount of liquid added is the same as that of the Lactobacillus plantarum liquid in Example 1. Other components, addition amounts and preparation methods are the same as those in Example 1.

[0091] Example 18

[0092] Compared with Example 2, the only difference between this embodiment is that the Lactobacillus plantarum liquid is replaced with the Lactobacillus fermentum liquid, and the amount of the liquid added is the same as that of the Lactobacillus plantarum liquid in Example 2. The other components, addition amounts and preparation methods are the same as those in Example 2.

[0093] Example 19

[0094] Compared with Example 3, the only difference between this embodiment is that the Lactobacillus plantarum liquid is replaced with the Lactobacillus fermentum liquid, and the amount of the liquid added is the same as that of the Lactobacillus plantarum liquid in Example 3. The other components, addition amounts and preparation methods are the same as those in Example 3.

[0095] Example 20

[0096] Compared with Example 4, the only difference between this embodiment is that the Lactobacillus plantarum liquid is replaced with Lactobacillus fermentum liquid, and the amount of liquid added is the same as that of the Lactobacillus plantarum liquid in Example 4. Other components, addition amounts and preparation methods are the same as those in Example 4.

[0097] Example 21

[0098] The only difference between this embodiment and Example 5 is that the Lactobacillus plantarum liquid is replaced with Lactobacillus fermentum liquid, and the amount of liquid added is the same as that of the Lactobacillus plantarum liquid in Example 5. The other components, addition amounts and preparation methods are the same as those in Example 5.

[0099] Example 22

[0100] Compared with Example 6, the only difference between this embodiment is that the Lactobacillus plantarum liquid is replaced with the Lactobacillus fermentum liquid, and the amount of the liquid added is the same as that of the Lactobacillus plantarum liquid in Example 6. The other components, addition amounts and preparation methods are the same as those in Example 6.

[0101] Example 23

[0102] Compared with Example 7, the only difference between this embodiment is that the Lactobacillus plantarum liquid is replaced with the Lactobacillus fermentum liquid, and the amount of the liquid added is the same as that of the Lactobacillus plantarum liquid in Example 7. The other components, addition amounts and preparation methods are the same as those in Example 7.

[0103] Example 24

[0104] The only difference between this embodiment and Example 8 is that the Lactobacillus plantarum liquid is replaced with Lactobacillus fermentum liquid, and the amount of liquid added is the same as that of the Lactobacillus plantarum liquid in Example 8. The other components, addition amounts and preparation methods are the same as those in Example 8.

[0105] Comparative Example 1 (the difference from Example 2 is only that conventional decoction is adopted)

[0106] 20 g of seabuckthorn fruit, 20 g of Glycyrrhiza inflata, 200 g of ethoxydiglycol, and 400 g of deionized water were mixed, and the mixture was heated and extracted at 100° C. for 2 h. The mixture was filtered and the filtrate was collected to obtain a plant extract.

[0107] Comparative Example 2 (the difference from Example 2 is only that ethoxydiglycol is replaced by propylene glycol)

[0108] The only difference between this comparative example and Example 2 is that ethoxydiglycol is replaced by propylene glycol, and the added amount is the same. The other components, addition amounts and preparation method are the same as those in Example 2.

[0109] Comparative Example 3 (the difference from Example 2 is that the plant lactobacillus liquid is replaced with deionized water)

[0110] Compared with Example 2, the only difference in this comparative example is that the Lactobacillus plantarum liquid is replaced with deionized water, and the added amount is the same. The other components, addition amounts and preparation method are the same as those in Example 2.

[0111] Comparative Example 4 (the only difference from Example 2 is that the plant lactobacillus liquid is replaced with saccharomyces cerevisiae liquid)

[0112] Compared with Example 2, the only difference in this comparative example is that the Lactobacillus plantarum liquid is replaced by Saccharomyces cerevisiae liquid, and the added amount is the same. Other components, addition amounts and preparation methods are the same as those in Example 2.

[0113] Comparative Example 5 (the difference from Example 2 is only that it lacks Glycyrrhiza inflata)

[0114] The preparation method of the plant fermentation product of this comparative example is as follows:

[0115] 1) 20 g of sea buckthorn fruit, 200 g of ethoxydiglycol, and 400 g of deionized water were mixed, and then added to a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0116] 2) 300g plant compound liquid, 6mL plant lactobacillus liquid (concentration of 10 8CFU / mL) and 4.5 g MRS medium were mixed and fermented at 28°C for 72 h. After the fermentation, the fermentation broth was sterilized at 121°C for 20 min, and the sterilized fermentation broth was centrifuged at 6000 rpm / min for 20 min. The supernatant after centrifugation was obtained to obtain a plant fermentation product.

[0117] Comparative Example 6 (the only difference from Example 2 is the lack of sea buckthorn fruit)

[0118] The preparation method of the plant fermentation product of this comparative example is as follows:

[0119] 1) 20 g of Glycyrrhiza inflata, 200 g of ethoxydiglycol, and 400 g of deionized water were mixed, and then added to a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite solution;

[0120] 2) 300g plant compound liquid, 6mL plant lactobacillus liquid (concentration of 10 8 CFU / mL) and 4.5 g MRS medium were mixed and fermented at 28°C for 72 h. After the fermentation, the fermentation broth was sterilized at 121°C for 20 min, and the sterilized fermentation broth was centrifuged at 6000 rpm / min for 20 min. The supernatant after centrifugation was obtained to obtain a plant fermentation product.

[0121] Test Example 1: Determination of total flavonoid content

[0122] This test example tests the total flavonoid content in the plant fermentation products of Examples 1-24 and the plant extracts or plant fermentation products of Comparative Examples 1-6 (hereinafter referred to as "samples"). The specific test method is as follows:

[0123] Adjust the total flavonoid concentration of the sample solution to the measurement range. Accurately pipette 2.0 mL of the adjusted sample solution into a dry, clean test tube. Add 0.2 mL of 5% sodium nitrite solution, shake well, and let stand for 5 minutes. Add 0.2 mL of 10% aluminum nitrate solution and shake well. After standing for 6 minutes, add 1.0 mL of 5% sodium hydroxide solution. Add 50% ethanol solution to the mark and shake the sample solution well. Rapidly measure the absorbance of each tube at a wavelength of 510 nm. Calculate the total flavonoid concentration from the sample absorbance according to the standard curve of rutin solution. The total flavonoid content in the sample is expressed as rutin.

[0124] Test Example 2: Determination of the contents of isorhamnetin, liquiritin, licoricechalcone A, glabridin and glycyrrhizic acid

[0125] This test example tests the content of isorhamnetin, liquiritin, licoricechalcone A, glabridin, and glycyrrhizic acid in the plant fermentation products of Examples 1-24 and the plant extracts or plant fermentation products of Comparative Examples 1-6 (hereinafter referred to as "samples"). The specific test method is as follows:

[0126] Preparation of test samples: Take 2.0 mL of sample and place it in a 10 mL volumetric flask. Add chromatographic grade methanol to the mark, shake well, and filter with a 0.45 μm filter membrane to obtain the test sample.

[0127] Chromatographic conditions: Agilent Poroshell 120EC-C column was used 18 (150mm×4.6mm, 2.7μm); gradient elution was performed with 0.05% phosphoric acid (A)-methanol (B) as the mobile phase (0min, 10% B; 3min, 25% B; 12min, 30% B; 20min, 40% B; 40min, 90% B; 50min, 95% B), flow rate 1.0mL / min; detection wavelength 270nm; column temperature 30℃.

[0128] The test results of Experimental Example 1 and Experimental Example 2 are shown in Table 1.

[0129] Table 1

[0130]

[0131]

[0132]

[0133] As can be seen from Table 1, compared with comparative examples 1-6, the total flavonoids in the plant fermentation products of Examples 1-24 are significantly improved. Isorhamnetin is the main flavonoid component in sea buckthorn fruit, and its content in the example samples is also higher than that in the comparative examples. The contents of flavonoid components liquiritin, glabridin and triterpenoid component glycyrrhizic acid derived from three types of licorice in the samples of Examples 1-24 are also higher than those in Comparative Examples 1-6. Licorice chalcone A is a characteristic flavonoid component in Glycyrrhiza inflata, and its content in the example samples using Glycyrrhiza inflata is higher than that in the corresponding comparative examples. The results of component analysis show that the preparation method provided by the present invention has a good extraction efficiency for flavonoids and triterpenes in sea buckthorn fruit and licorice, and the obtained plant fermentation has a higher content of active ingredients.

[0134] Experimental Example 3: Cell scratch test

[0135] This test example evaluated the cell repair ability and the effect on cell migration of the plant fermented products of Examples 1-24 and the plant extracts or plant fermented products of Comparative Examples 1-6 (hereinafter referred to as "samples"). The specific test methods are as follows:

[0136] Use a pen to evenly draw horizontal lines on the back of a 6-well plate, with intervals of 0.5-1 cm between the lines. Fill the wells with human immortalized epidermal cells (HaCaT). The next day, once the cells have filled the wells, use a pipette tip to scratch the wells along the pre-drawn horizontal lines on the 6-well plate. Wash the cells three times with PBS, remove the scratched cells, add cell culture medium containing 1% (w / w) sample, and incubate in a 37°C, 5% CO2 incubator for 24 hours. Take photos, and calculate the percentage of wound healing after 24 hours. Deionized water was used instead of sample to serve as a blank control group. The evaluation results are shown in Table 2.

[0137] Table 2

[0138]

[0139]

[0140] As shown in Table 2, compared with Comparative Examples 1 to 6, the samples of Examples 1 to 24 can more effectively enhance cell migration and repair capabilities. The results show that the plant fermentation product of the present invention has a good repair effect.

[0141] Experimental Example 4: Effects on IL-6 and TNF-α Contents in HaCaT Cells after UV Irradiation

[0142] HaCat cells were seeded into 6-well plates and incubated overnight in an incubator at 37°C and 5% CO2. When the cell plating rate in the 6-well plate reached 50%-60%, the old culture medium was removed. A blank control group, a model group, and a sample group were set up. The blank control group and the model group were added with culture medium, 100 μL per well; the sample group was added with cell culture medium containing 1 wt% sample, 100 μL per well, and pretreated for 1 hour. The model group and the sample group were exposed to 30 mJ / cm 2 The cells were irradiated with UVB for 40 minutes and then cultured in an incubator at 37°C and 5% CO2 for 24 hours. After the culture period, the IL-6 and TNF-α levels were determined according to the ELISA kit protocol. The test results are shown in Table 3.

[0143] Table 3

[0144]

[0145]

[0146]

[0147] As shown in Table 3, compared with the model group, the samples of Examples 1-24 and Comparative Examples 1-6 were able to reduce the production of IL-6 and TNF-α. However, compared with the samples of Comparative Examples 1-6, the samples of Examples 1-24 had a stronger inhibitory effect on the production of inflammatory factors, indicating that the plant fermentation products of the present invention have a stronger soothing effect.

[0148] Test Example 5: Effects on Staphylococcus aureus and Malassezia

[0149] Staphylococcus aureus is a common skin pathogen that typically causes papules or pustules. Propionibacterium acnes is also a common skin pathogen that can induce acne, causing small, red, raised nodules to form on the skin surface, accompanied by itching and other skin problems.

[0150] Preparation of bacterial suspension: Staphylococcus aureus (ATCC6538) and Propionibacterium acnes (ATCC6919) were selected as test bacteria. The test bacteria were prepared into bacterial suspension with PBS at a concentration of 5×10 6 CFU / mL.

[0151] Inhibition zone test: The inhibition zone test was conducted using the Oxford cup method. 100 μL of the bacterial suspension was pipetted onto the surface of the solidified culture medium and evenly spread with a sterile applicator. The plate was covered and incubated at 20°C for 10 minutes. Finally, the sterile Oxford cup was gently inverted on the surface of the culture medium. Sample solutions of varying mass fractions were added dropwise until the liquid level was parallel to the cup surface. The plate was then incubated at 37°C for 18 hours. Inhibition zone results were determined according to the company standard "Microbial Antimicrobial Evaluation In Vitro Inhibition of Bacteria and Fungi (Drug Susceptibility Testing)" as follows: an inhibition zone diameter >20 mm indicates extreme sensitivity (denoted as ++++); 15-20 mm indicates high sensitivity (denoted as +++); 10-15 mm indicates moderate sensitivity (denoted as ++); <10 mm indicates low sensitivity (denoted as +); and 0 mm indicates no inhibition (denoted as -). The test results are shown in Table 4.

[0152] Table 4

[0153]

[0154]

[0155] The test results in Table 4 show that the samples of Examples 1-24 exhibited inhibitory effects against Staphylococcus aureus and Propionibacterium acnes at either 7.5% or 2.0% mass fraction. Compared to the samples of Comparative Examples 1-6 at equivalent concentrations, the samples exhibited stronger inhibitory effects against Staphylococcus aureus and Propionibacterium acnes. The plant fermentation products of the present invention exhibited a strong inhibitory effect against pathogens, demonstrating their potential for improving inflammation caused by common pathogens and maintaining a healthy skin microbiome.

[0156] Application Example 1

[0157] An emulsion comprising the plant fermentation product prepared in Example 2 of the present invention. The specific formula is shown in Table 5. The preparation method thereof can be prepared according to the conventional preparation method of emulsion.

[0158] Table 5

[0159]

[0160]

[0161] Application Example 2

[0162] The only difference between the emulsion of Application Example 2 and Application Example 1 is that the plant fermentation product prepared in Example 2 is replaced by the plant fermentation product prepared in Example 8. Other components, addition amounts and preparation methods are the same as those of Application Example 1.

[0163] Comparative Application Example 1

[0164] The difference between the emulsion of Application Example 1 and Application Example 1 is that the plant fermentation product of Example 2 is replaced by the plant extract of Comparative Example 1 in Application Example 1, and the other ingredients, dosages and preparation methods are the same as those in Application Example 1.

[0165] Blank control group

[0166] The difference between the emulsion in the blank control group and that in Application Example 1 is that the plant fermentation product prepared in Example 2 is replaced by deionized water, and the other components, addition amounts and preparation methods are the same as those in Application Example 1.

[0167] The whitening and soothing efficacy of the emulsions of Application Example 1, Application Example 2, Comparative Application Example 1 and the blank control group were evaluated.

[0168] 40 volunteers aged 20-45, half male and half female, were selected as subjects. They were divided into 4 groups and used the emulsions of Application Example 1, Application Example 2, Comparative Application Example 1 and the blank control group respectively. The test site was the face. After cleansing the face every morning and evening, the subjects took an appropriate amount of the product and pressed it evenly on the face, massaging until it was absorbed. Use it twice a day, once in the morning and once in the evening. The subjects were followed up before using the product (day 0), 14 days after using the product, and 28 days after using the product. The evaluation indicators include skin erythema and desquamation at the test site. Each indicator is evaluated according to its degree with a score of 0 (very good) to 9 points (very poor). In addition, the repair and soothing effect of the product was evaluated by comparing the skin moisture content, transepidermal water loss (TEWL) and skin red area before and after use of the product. The results are shown in Table 6.

[0169] Table 6

[0170]

[0171]

[0172] As can be seen from Table 6, compared with the emulsion of comparative application example 1 and the blank control group, the subjects believed that skin erythema and desquamation were improved after using the emulsions of application example 1 and application example 2; combined with the changes in stratum corneum water content, TEWL value and red zone area before and after use by the subjects, it was shown that the plant fermentation of the present invention has significant soothing and repairing functions.

[0173] The above specifically describes the preferred embodiments of the present invention, but the invention is not limited to the embodiments. Those skilled in the art may make various equivalent modifications or substitutions without violating the spirit of the present invention. These equivalent modifications or substitutions are all included in the scope defined by the claims of this application.

Claims

1. A plant fermentation product, characterized in that: The raw materials for preparing the plant fermentation product include: plant raw materials, solvents, and fermentation strains; the plant raw materials are sea buckthorn fruit and liquorice; the solvents include solvent A and solvent B, the solvent A includes glycerol and / or ethoxydiglycol, and the solvent B includes water; the fermentation strain is at least one of Lactobacillus plantarum, Lactobacillus casei, and Lactobacillus fermentum; The usage ratio of the seabuckthorn fruit, the liquorice and the solvent is (0.001-0.1g): (0.001-0.1g): 1g; The usage ratio of the solvent A and the solvent B is (0.01-5g):1g; The plant fermentation product is prepared by a preparation method comprising the following steps: Mixing the plant raw material and the solvent, and homogenizing under high pressure to obtain a plant composite liquid; The plant complex liquid and the fermentation strain are mixed and fermented to obtain the plant fermentation product.

2. The plant fermentation product according to claim 1, characterized in that The licorice comprises at least one of Glycyrrhiza glabra, Glycyrrhiza inflata and Glycyrrhiza uralensis.

3. The plant fermentation product according to claim 1, characterized in that The concentration of the fermentation strain is 10 5 -10 10 CFU / mL.

4. The plant fermentation product according to claim 1, characterized in that Before the fermentation, the method further includes adding a culture medium; the usage ratio of the plant complex liquid, the fermentation strain and the culture medium is (1-500 g): (0.01-10 mL): 1 g.

5. The plant fermentation product according to claim 1, characterized in that The pressure of the high-pressure homogenization is 10-100 MPa; and / or, the temperature of the high-pressure homogenization is 25-50° C.; and / or, the time of the high-pressure homogenization is 5-40 min.

6. The plant fermentation product according to claim 1, characterized in that The fermentation temperature is 20-45° C.; and / or the fermentation time is 12-120 h.

7. A cosmetic, characterized in that: The invention comprises the plant fermentation product according to any one of claims 1 to 6.

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