Preparation method and application of rosa roxburghii fermented product

Abstract

The invention relates to the technical field of daily chemicals, in particular to a preparation method and application of a rosa roxburghii fermented product. The preparation method comprises the following steps: S01, taking fresh roxburgh rose fruits, cleaning with clear water, airing, adding a solution, crushing, homogenizing, and pouring into a sealed container; s02, then adding a culture solution A and a strain A for fermentation; after the fermentation is finished, filtering a fermentation product; performing freezing; and S03, naturally melting the fermentation product obtained in the step S02 at room temperature, adding a culture medium B for continuous fermentation, filtering to obtain the roxburgh rose fermentation product, and refrigerating and storing. The rosa roxburghii tratt leavening and related products provided by the invention have good whitening, anti-wrinkle, soothing, anti-aging and other effects, the active ingredients are natural and green, and the safety is high.

Description

Technical Field

[0001] The invention relates to the technical field of daily chemicals, and in particular to a preparation method and application of a roxburghii fermented product. Background Art

[0002] In recent years, people have become more and more strict about the efficacy of cosmetics. Traditional cosmetic ingredients are often accompanied by certain side effects, such as chemical residues, skin allergic reactions, etc. These problems have gradually caused consumers' concerns and attention. The fruit of the roxburghii is rich in VC, SOD, organic acids, polysaccharides, flavonoids and polyphenols, and a variety of trace elements. It has skin care effects such as antioxidant, anti-aging, whitening, and antibacterial. The roxburghii is a plant with both medicinal and edible properties. Its functional ingredients are natural, mild and non-irritating. Compared with ordinary extraction, fermentation technology can decompose large molecular active substances into small molecules that are easier for the skin to absorb, thereby improving the performance of the roxburghii in cosmetics.

[0003] Rosa roxburghii is rich in vitamins, trace elements, amino acids, higher fatty acids and polyacids. Rosa roxburghii extract also has excellent antioxidant, anti-aging and spot-lightening effects.

[0004] CN117070586A A composite polypeptide of roxburghii and its preparation method and application, belonging to the field of fermentation technology. The composite polypeptide is added with shellfish, melons, beans, and wheat on the basis of roxburghii fruit to increase protein group, vitamin group, and mineral group nutrients, and roxburghii fruit is used as fermentation base material and bifidobacterium and lactic acid bacteria are used as fermentation sources through cultivation, inactivation and decomposition to obtain a composite polypeptide of roxburghii formed by metabolites, cell lipid micromolecule fragments, cell wall components and polysaccharide complexes, i.e., bifid yeast fermentation product lysate, and a large number of peptide polypeptide groups, bifid yeast, amino acid yeast peptide groups, superoxide dismutase and other fermentation clusters, trace element clusters, mineral enzyme clusters, vitamin C high content group and other small molecule clusters in the fermented filtrate and filter residue, producing more extensive small molecule biological activities, which can be used for skin repair, skin nourishment, slowing down cell aging, and enhancing skin quality antibodies, and has a great promotion prospect in the field of cosmetics and food.

[0005] The above patents do not solve the problem that the roxburghii is easy to change color, and the antioxidant properties of the roxburghii after the color change are also greatly reduced. In view of this, the present invention provides a preparation method and application of a roxburghii fermented product. Summary of the invention

[0006] In order to solve the above technical problems, the present invention provides a method for preparing a roxburghii fermented product, comprising the steps of:

[0007] S01 Take fresh roxburghii fruit, wash it with clean water and dry it, add solution to crush it and homogenize it, and pour it into a sealed container;

[0008] S02 then adds culture solution A and strain A to carry out fermentation; after the fermentation is completed, the fermentation product is filtered and frozen;

[0009] S03: naturally thaw the fermented product obtained in step S02 at room temperature, add culture medium B to continue fermentation, filter to obtain the roxburghii fermented product, and store in a refrigerator.

[0010] As an embodiment of the present invention, the solution in step S01 is a fructose aqueous solution.

[0011] As an embodiment of the present invention, the component of the culture solution A in S02 is 1000 mL of wort.

[0012] As an embodiment of the present invention, the bacterial species A in S02 is yeast and bifidobacterium.

[0013] As an embodiment of the present invention, the components of the culture medium B in S03 are 10.0g beef extract powder, 5.0g peptone, 3.0g yeast extract powder, 5.0g glucose, 1.0g soluble starch, 5.0g NaCl, 3.0g NaAc, 0.5g L-cysteine ​​hydrochloride, and 1000.0mL distilled water.

[0014] The second aspect of the present invention provides an essence, the raw materials for preparing the essence include the roxburghii fermented product.

[0015] As an embodiment of the present invention, the essence comprises: deionized water, butylene glycol, glycerin, polyacrylate cross-linked polymer-6, the roxburghii fermented product according to any one of claims 1 to 5, p-hydroxyacetophenone, 1,2-hexanediol, and disodium EDTA.

[0016] The third aspect of the invention provides a method for preparing the essence, comprising the following steps:

[0017] S01 Stir glycerol and p-hydroxyacetophenone in a water bath at 70°C and dissolve completely;

[0018] S02: add deionized water into the container, add butanediol and polyacrylate cross-linked polymer-6, raise the temperature to 80-85°C, keep warm and stir at 800rpm-1000rpm for 10min-20min until uniform; when the material body is cooled to 45-50°C, add the mixture obtained in S01 and stir evenly; continue to cool the material body to 30-35°C, add the sea buckthorn fermentation product, 1,2-hexanediol and disodium EDTA in sequence, and stir evenly.

[0019] The fourth aspect of the invention provides the roxburghii fermented product, characterized in that the roxburghii fermented product is used in the field of skin care.

[0020] The fifth aspect of the invention provides the roxburghii fermented product, which is used in the fields of moisturizing and anti-aging.

[0021] By adopting the above technical solution, the present invention has the following beneficial effects:

[0022] The roxburghii fermented product and related products provided by the invention have good whitening, anti-wrinkle, soothing, anti-aging and other effects, and the active ingredients are of natural green sources and are highly safe. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings in the following description are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0024] Figure 1 Photo of the serum of Example 1 DETAILED DESCRIPTION

[0025] The technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0026] The present invention provides a method for preparing a roxburghii fermented product, the steps comprising:

[0027] S01 Take fresh roxburghii fruit, wash it with clean water and dry it, add solution to crush it and homogenize it, and pour it into a sealed container;

[0028] The mass ratio of the roxburghii fruit to the solution is 1:1.5; the solution is a 1wt% fructose aqueous solution;

[0029] S02 then adds culture solution A and strain A to carry out fermentation; after the fermentation is completed, the fermentation product is filtered and frozen;

[0030] The culture solution A is 1000 ml of malt juice, the mass ratio of the roxburghii slurry to the culture solution A is 1:10; the mass ratio of the strain A to the culture solution A is 3:100; the strain A contains yeast and bifidobacterium; and the inoculation ratio is 1:2.

[0031] The yeast is Rhodosporidium toruloides, which is from the China Industrial Microbiological Culture Collection Center.

[0032] The bifidobacterium is Bifidobacterium animalis subsp. lactis, which is from the China Industrial Microbiological Culture Collection Center.

[0033] The fermentation conditions are sealed fermentation, fermentation temperature 30° C., and fermentation time 24 hours.

[0034] The filtration was clarified by using a 0.1um filter membrane; the freezing was cryogenically frozen by liquid nitrogen.

[0035] S03: naturally thaw the fermented product obtained in step S02 at room temperature, add culture medium B to continue fermentation, filter to obtain the roxburghii fermented product, and store in a refrigerator.

[0036] The components of the culture medium B in the S03 are 10.0 g beef extract powder, 5.0 g peptone, 3.0 g yeast extract powder, 5.0 g glucose, 1.0 g soluble starch, 5.0 g NaCl, 3.0 g NaAc, 0.5 g L-cysteine ​​hydrochloride, and 1000.0 mL distilled water.

[0037] The fermentation temperature is 30° C., the fermentation time is 12 hours, and after the fermentation is completed, 0.1 μm filter membrane is used to filter and clarify to obtain the roxburghii fermented product.

[0038] As a preferred embodiment of the present invention, 1 wt % chrysanthemum powder is added to the roxburghii fermented product during the S03 fermentation.

[0039] The roxburghii fermented product obtained by the invention has small molecules, is easy to absorb, has a relatively light color, and is not easy to change color.

[0040] The present invention is further explained below in conjunction with specific implementation modes.

[0041] Example 1

[0042] This embodiment provides an essence liquid, and its preparation raw materials include

[0043] Deionized water 88.35%, butylene glycol 4%, glycerin 3%, polyacrylate cross-linked polymer-6 0.6%, roxburghii fermentation product 3%, p-hydroxyacetophenone 0.5%, 1,2-hexanediol 0.5%, disodium EDTA 0.05%.

[0044] The preparation method of the roxburghii fermented product in this embodiment is as follows:

[0045] S01 takes fresh roxburghii fruit, washes it with clean water and then dries it, adds solution to crush it and homogenizes it, and then pours it into a sealed container to obtain roxburghii slurry;

[0046] The mass ratio of the roxburghii fruit to the solution is 1:1.5; the solution is a 1wt% fructose aqueous solution;

[0047] S02 then adds culture solution A and strain A to carry out fermentation; after the fermentation is completed, the fermentation product is filtered and frozen;

[0048] The culture solution A is 1000 ml of malt juice, the mass ratio of the roxburghii slurry to the culture solution A is 1:10; the mass ratio of the strain A to the culture solution A is 3:100; the strain A contains yeast and bifidobacterium; and the inoculation ratio is 1:2.

[0049] The yeast is Rhodosporidium toruloides, which is from the China Industrial Microbiological Culture Collection Center.

[0050] The bifidobacterium is Bifidobacterium animalis subsp. lactis, which is from the China Industrial Microbiological Culture Collection Center.

[0051] The fermentation conditions are sealed fermentation, and the total volume of the fermentation liquid is 2 / 3 of the total volume of the sealed container; the fermentation temperature is 30° C., and the fermentation time is 24 hours.

[0052] The filtration was clarified by using a 0.1um filter membrane; the freezing was cryogenically frozen by liquid nitrogen.

[0053] S03: the fermented product obtained in step S02 is naturally melted at room temperature, and the fermented product and chrysanthemum powder are added to the culture medium B to continue fermentation, and the fermented product of roxburghii is filtered and refrigerated for storage.

[0054] The mass ratio of the fermented product obtained in S02 to the culture medium B is 1:10; the mass ratio of the chrysanthemum powder to the culture medium B is 1:100.

[0055] The components of the culture medium B in the S03 are 10.0 g beef extract powder, 5.0 g peptone, 3.0 g yeast extract powder, 5.0 g glucose, 1.0 g soluble starch, 5.0 g NaCl, 3.0 g NaAc, 0.5 g L-cysteine ​​hydrochloride, and 1000.0 mL distilled water.

[0056] The fermentation temperature is 30° C., the fermentation time is 12 hours, and after the fermentation is completed, 0.1 μm filter membrane is used to filter and clarify to obtain the roxburghii fermented product.

[0057] The preparation method of the essence comprises the following steps: pre-preparation phase:

[0058] S01 Stir glycerol and p-hydroxyacetophenone in a water bath at 70°C and dissolve completely;

[0059] S02: add deionized water into the container, add butanediol and polyacrylate cross-linked polymer-6, raise the temperature to 80-85°C, keep warm and stir at 800rpm-1000rpm for 10min-20min until uniform; when the material body is cooled to 45-50°C, add the mixture obtained in S01 and stir evenly; continue to cool the material body to 30-35°C, add the sea buckthorn fermentation product, 1,2-hexanediol and disodium EDTA in sequence, and stir evenly.

[0060] Example 2

[0061] This embodiment provides an essence liquid, and its preparation raw materials include

[0062] Deionized water 88.35%, butylene glycol 4%, glycerin 3%, polyacrylate cross-linked polymer-6 0.6%, roxburghii fermentation product 3%, p-hydroxyacetophenone 0.5%, 1,2-hexanediol 0.5%, disodium EDTA 0.05%.

[0063] The preparation method of the roxburghii fermented product in this embodiment is as follows:

[0064] S01 takes fresh roxburghii fruit, washes it with clean water and then dries it, adds solution to crush it and homogenizes it, and then pours it into a sealed container to obtain roxburghii slurry;

[0065] The mass ratio of the roxburghii fruit to the solution is 1:1.5; the solution is a 1wt% fructose aqueous solution;

[0066] S02 then adds culture solution A and strain A to carry out fermentation; after the fermentation is completed, the fermentation product is filtered and frozen;

[0067] The culture solution A is 1000 ml of malt juice, the mass ratio of the roxburghii slurry to the culture solution A is 1:10; the mass ratio of the strain A to the culture solution A is 3:100; the strain A contains yeast and bifidobacterium; and the inoculation ratio is 1:2.

[0068] The yeast is Rhodosporidium toruloides, which is from the China Industrial Microbiological Culture Collection Center.

[0069] The bifidobacterium is Bifidobacterium animalis subsp. lactis, which is from the China Industrial Microbiological Culture Collection Center.

[0070] The fermentation conditions are sealed fermentation, and the total volume of the fermentation liquid is 2 / 3 of the total volume of the sealed container; the fermentation temperature is 30° C., and the fermentation time is 24 hours.

[0071] The filtration was clarified by using a 0.1um filter membrane; the freezing was cryogenically frozen by liquid nitrogen.

[0072] S03: naturally thaw the fermented product obtained in step S02 at room temperature, add culture medium B to continue fermentation, filter to obtain the roxburghii fermented product, and store in a refrigerator.

[0073] The mass ratio of the fermentation product obtained in S02 to the culture medium B is 1:10.

[0074] The components of the culture medium B in the S03 are 10.0 g beef extract powder, 5.0 g peptone, 3.0 g yeast extract powder, 5.0 g glucose, 1.0 g soluble starch, 5.0 g NaCl, 3.0 g NaAc, 0.5 g L-cysteine ​​hydrochloride, and 1000.0 mL distilled water.

[0075] The fermentation temperature is 30° C., the fermentation time is 12 hours, and after the fermentation is completed, 0.1 μm filter membrane is used to filter and clarify to obtain the roxburghii fermented product.

[0076] The preparation method of the essence comprises the following steps: pre-preparation phase:

[0077] S01 Stir glycerol and p-hydroxyacetophenone in a water bath at 70°C and dissolve completely;

[0078] S02: add deionized water into the container, add butanediol and polyacrylate cross-linked polymer-6, raise the temperature to 80-85°C, keep warm and stir at 800rpm-1000rpm for 10min-20min until uniform; when the material body is cooled to 45-50°C, add the mixture obtained in S01 and stir evenly; continue to cool the material body to 30-35°C, add the sea buckthorn fermentation product, 1,2-hexanediol and disodium EDTA in sequence, and stir evenly.

[0079] Example 3

[0080] This embodiment provides an essence liquid, and its preparation raw materials include

[0081] Deionized water 88.35%, butylene glycol 4%, glycerin 3%, polyacrylate cross-linked polymer-6 0.6%, roxburghii fermentation product 3%, p-hydroxyacetophenone 0.5%, 1,2-hexanediol 0.5%, disodium EDTA 0.05%.

[0082] The preparation method of the roxburghii fermented product in this embodiment is as follows:

[0083] S01 takes fresh roxburghii fruit, washes it with clean water and then dries it, adds water to crush it and homogenizes it, and then pours it into a sealed container to obtain roxburghii slurry;

[0084] The mass ratio of the roxburghii fruit to water is 1:1.5;

[0085] S02 then adds culture solution A and strain A to carry out fermentation; after the fermentation is completed, the fermentation product is filtered and frozen;

[0086] The culture solution A is 1000 ml of malt juice, the mass ratio of the roxburghii slurry to the culture solution A is 1:10; the mass ratio of the strain A to the culture solution A is 3:100; the strain A contains yeast and bifidobacterium; and the inoculation ratio is 1:2.

[0087] The yeast is Rhodosporidium toruloides, which is from the China Industrial Microbiological Culture Collection Center.

[0088] The bifidobacterium is Bifidobacterium animalis subsp. lactis, which is from the China Industrial Microbiological Culture Collection Center.

[0089] The fermentation conditions are sealed fermentation, and the total volume of the fermentation liquid is 2 / 3 of the total volume of the sealed container; the fermentation temperature is 30° C., and the fermentation time is 24 hours.

[0090] The filtration was clarified by using a 0.1um filter membrane; the freezing was cryogenically frozen by liquid nitrogen.

[0091] S03: the fermented product obtained in step S02 is naturally melted at room temperature, and the fermented product and chrysanthemum powder are added to the culture medium B to continue fermentation, and the fermented product of roxburghii is filtered and refrigerated for storage.

[0092] The mass ratio of the fermented product obtained in S02 to the culture medium B is 1:10; the mass ratio of the chrysanthemum powder to the culture medium B is 1:100.

[0093] The components of the culture medium B in the S03 are 10.0 g beef extract powder, 5.0 g peptone, 3.0 g yeast extract powder, 5.0 g glucose, 1.0 g soluble starch, 5.0 g NaCl, 3.0 g NaAc, 0.5 g L-cysteine ​​hydrochloride, and 1000.0 mL distilled water.

[0094] The fermentation temperature is 30° C., the fermentation time is 12 hours, and after the fermentation is completed, 0.1 μm filter membrane is used to filter and clarify to obtain the roxburghii fermented product.

[0095] The preparation method of the essence comprises the following steps: pre-preparation phase:

[0096] S01 Stir glycerol and p-hydroxyacetophenone in a water bath at 70°C and dissolve completely;

[0097] S02: add deionized water into the container, add butanediol and polyacrylate cross-linked polymer-6, raise the temperature to 80-85°C, keep warm and stir at 800rpm-1000rpm for 10min-20min until uniform; when the material body is cooled to 45-50°C, add the mixture obtained in S01 and stir evenly; continue to cool the material body to 30-35°C, add the sea buckthorn fermentation product, 1,2-hexanediol and disodium EDTA in sequence, and stir evenly.

[0098] Example 4

[0099] This embodiment provides an essence liquid, and its preparation raw materials include

[0100] Deionized water 88.35%, butylene glycol 4%, glycerin 3%, polyacrylate crosspolymer-6 0.6%, roxburghii fermentation product 3%, p-hydroxyacetophenone 0.5%, 1,2-hexanediol 0.5%, disodium EDTA 0.05%.

[0101] The preparation method of the roxburghii fermented product in this embodiment is as follows:

[0102] S01 takes fresh roxburghii fruit, washes it with clean water and then dries it, adds solution to crush it and homogenizes it, and then pours it into a sealed container to obtain roxburghii slurry;

[0103] The mass ratio of the roxburghii fruit to the solution is 1:1.5; the solution is a 1wt% fructose aqueous solution;

[0104] S02 then adds culture solution A and strain A to carry out fermentation; after the fermentation is completed, the fermentation product is filtered;

[0105] The culture solution A is 1000 ml of malt juice, the mass ratio of the roxburghii slurry to the culture solution A is 1:10; the mass ratio of the strain A to the culture solution A is 3:100; the strain A contains yeast and bifidobacterium; and the inoculation ratio is 1:2.

[0106] The yeast is Rhodosporidium toruloides, which is from the China Industrial Microbiological Culture Collection Center.

[0107] The bifidobacterium is Bifidobacterium animalis subsp. lactis, which is from the China Industrial Microbiological Culture Collection Center.

[0108] The fermentation conditions are sealed fermentation, and the total volume of the fermentation liquid is 2 / 3 of the total volume of the sealed container; the fermentation temperature is 30° C., and the fermentation time is 24 hours.

[0109] Filtration was performed using a 0.1 um filter membrane for clarification.

[0110] S03: adding the fermented product obtained in step S02 and the chrysanthemum powder into the culture medium B to continue fermentation, filtering to obtain the roxburghii fermented product, and refrigerating and storing.

[0111] The mass ratio of the fermented product obtained in S02 to the culture medium B is 1:10; the mass ratio of the chrysanthemum powder to the culture medium B is 1:100.

[0112] The components of the culture medium B in the S03 are 10.0 g beef extract powder, 5.0 g peptone, 3.0 g yeast extract powder, 5.0 g glucose, 1.0 g soluble starch, 5.0 g NaCl, 3.0 g NaAc, 0.5 g L-cysteine ​​hydrochloride, and 1000.0 mL distilled water.

[0113] The fermentation temperature is 30° C., the fermentation time is 12 hours, and after the fermentation is completed, 0.1 μm filter membrane is used to filter and clarify to obtain the roxburghii fermented product.

[0114] The preparation method of the essence comprises the following steps: pre-preparation phase:

[0115] S01 Stir glycerol and p-hydroxyacetophenone in a water bath at 70°C and dissolve completely;

[0116] S02: add deionized water into the container, add butanediol and polyacrylate cross-linked polymer-6, raise the temperature to 80-85°C, keep warm and stir at 800rpm-1000rpm for 10min-20min until uniform; when the material body is cooled to 45-50°C, add the mixture obtained in S01 and stir evenly; continue to cool the material body to 30-35°C, add the sea buckthorn fermentation product, 1,2-hexanediol and disodium EDTA in sequence, and stir evenly.

[0117] Example 5

[0118] This embodiment provides an essence liquid, and its preparation raw materials include

[0119] Deionized water 88.35%, butylene glycol 4%, glycerin 3%, polyacrylate cross-linked polymer-6 0.6%, roxburghii fermentation product 3%, p-hydroxyacetophenone 0.5%, 1,2-hexanediol 0.5%, disodium EDTA 0.05%.

[0120] The preparation method of the roxburghii fermented product in this embodiment is as follows:

[0121] S01 takes fresh roxburghii fruit, washes it with clean water and then dries it, adds solution to crush it and homogenizes it, and then pours it into a sealed container to obtain roxburghii slurry;

[0122] The mass ratio of the roxburghii fruit to the solution is 1:1.5; the solution is a 1wt% fructose aqueous solution;

[0123] S02 then adds culture medium A and strain A to ferment; after the fermentation is completed, the fermentation product is filtered and refrigerated for storage.

[0124] The culture solution A is 1000 ml of malt juice, the mass ratio of the roxburghii slurry to the culture solution A is 1:10; the mass ratio of the strain A to the culture solution A is 3:100; the strain A contains yeast and bifidobacterium; and the inoculation ratio is 1:2.

[0125] The yeast is Rhodosporidium toruloides, which is from the China Industrial Microbiological Culture Collection Center.

[0126] The bifidobacterium is Bifidobacterium animalis subsp. lactis, which is from the China Industrial Microbiological Culture Collection Center.

[0127] The fermentation conditions are sealed fermentation, and the total volume of the fermentation liquid is 2 / 3 of the total volume of the sealed container; the fermentation temperature is 30° C., and the fermentation time is 24 hours.

[0128] Filtration was performed using a 0.1 um filter membrane for clarification.

[0129] The mass ratio of the fermented product obtained in S02 to the culture medium B is 1:10; the mass ratio of the chrysanthemum powder to the culture medium B is 1:100.

[0130] The preparation method of the essence comprises the following steps: pre-preparation phase:

[0131] S01 Stir glycerol and p-hydroxyacetophenone in a water bath at 70°C and dissolve completely;

[0132] S02: add deionized water into the container, add butanediol and polyacrylate cross-linked polymer-6, raise the temperature to 80-85°C, keep warm and stir at 800rpm-1000rpm for 10min-20min until uniform; when the material body is cooled to 45-50°C, add the mixture obtained in S01 and stir evenly; continue to cool the material body to 30-35°C, add the sea buckthorn fermentation product, 1,2-hexanediol and disodium EDTA in sequence, and stir evenly.

[0133] Example 6

[0134] This embodiment provides an essence liquid, and its preparation raw materials include

[0135] Deionized water 88.35%, butylene glycol 4%, glycerin 3%, polyacrylate cross-linked polymer-6 0.6%, roxburghii fermentation product 3%, p-hydroxyacetophenone 0.5%, 1,2-hexanediol 0.5%, disodium EDTA 0.05%.

[0136] The preparation method of the roxburghii fermented product in this embodiment is as follows:

[0137] S01 takes fresh roxburghii fruit, washes it with clean water and then dries it, adds solution to crush it and homogenizes it, and then pours it into a sealed container to obtain roxburghii slurry;

[0138] The mass ratio of the roxburghii fruit to the solution is 1:1.5; the solution is a 1wt% fructose aqueous solution;

[0139] S02 then adds culture medium A and strain A for fermentation; after the fermentation is completed, the fermented product is filtered and refrigerated for storage.

[0140] The culture solution A is 1000 ml of malt juice, the mass ratio of the roxburghii slurry to the culture solution A is 1:10; the mass ratio of the strain A to the culture solution A is 3:100; the strain A contains yeast and bifidobacterium; and the inoculation ratio is 1:2.

[0141] The yeast is Rhodosporidium toruloides, which is from the China Industrial Microbiological Culture Collection Center.

[0142] The bifidobacterium is Bifidobacterium animalis subsp. lactis, which is from the China Industrial Microbiological Culture Collection Center.

[0143] The fermentation conditions are sealed fermentation, and the total volume of the fermentation liquid is 2 / 3 of the total volume of the sealed container; the fermentation temperature is 30° C., and the fermentation time is 36 hours.

[0144] Filtration was performed using a 0.1 um filter membrane for clarification.

[0145] The mass ratio of the fermented product obtained in S02 to the culture medium B is 1:10; the mass ratio of the chrysanthemum powder to the culture medium B is 1:100.

[0146] The preparation method of the essence comprises the following steps: pre-preparation phase:

[0147] S01 Stir glycerol and p-hydroxyacetophenone in a water bath at 70°C and dissolve completely;

[0148] S02: add deionized water into the container, add butanediol and polyacrylate cross-linked polymer-6, raise the temperature to 80-85°C, keep warm and stir at 800rpm-1000rpm for 10min-20min until uniform; when the material body is cooled to 45-50°C, add the mixture obtained in S01 and stir evenly; continue to cool the material body to 30-35°C, add the sea buckthorn fermentation product, 1,2-hexanediol and disodium EDTA in sequence, and stir evenly.

[0149] Example 7

[0150] This embodiment provides an essence liquid, and its preparation raw materials include

[0151] Deionized water 88.35%, butylene glycol 4%, glycerin 3%, polyacrylate cross-linked polymer-6 0.6%, roxburghii fermentation product 3%, p-hydroxyacetophenone 0.5%, 1,2-hexanediol 0.5%, disodium EDTA 0.05%.

[0152] The preparation method of the roxburghii fermented product in this embodiment is as follows:

[0153] S01 takes fresh roxburghii fruit, washes it with clean water and then dries it, adds solution to crush it and homogenizes it, and then pours it into a sealed container to obtain roxburghii slurry;

[0154] The mass ratio of the roxburghii fruit to the solution is 1:1.5; the solution is a 1wt% fructose aqueous solution;

[0155] S02 then adds culture solution A and strain A to carry out fermentation; after the fermentation is completed, the fermentation product is filtered and frozen;

[0156] The culture solution A is 1000 ml of malt juice, the mass ratio of the roxburghii slurry to the culture solution A is 1:10; the mass ratio of the strain A to the culture solution A is 3:100; the strain A is a yeast. The yeast is Rhodosporidium toruloides, which is from the China Industrial Microbiological Culture Collection Administration Center.

[0157] The fermentation conditions are sealed fermentation, and the total volume of the fermentation liquid is 2 / 3 of the total volume of the sealed container; the fermentation temperature is 30° C., and the fermentation time is 24 hours.

[0158] The filtration was clarified by using a 0.1um filter membrane; the freezing was cryogenically frozen by liquid nitrogen.

[0159] S03: the fermented product obtained in step S02 is naturally melted at room temperature, and the fermented product and chrysanthemum powder are added to the culture medium B to continue fermentation, and the fermented product of roxburghii is filtered and refrigerated for storage.

[0160] The mass ratio of the fermented product obtained in S02 to the culture medium B is 1:10; the mass ratio of the chrysanthemum powder to the culture medium B is 1:100.

[0161] The components of the culture medium B in the S03 are 10.0 g beef extract powder, 5.0 g peptone, 3.0 g yeast extract powder, 5.0 g glucose, 1.0 g soluble starch, 5.0 g NaCl, 3.0 g NaAc, 0.5 g L-cysteine ​​hydrochloride, and 1000.0 mL distilled water.

[0162] The fermentation temperature is 30° C., the fermentation time is 12 hours, and after the fermentation is completed, 0.1 μm filter membrane is used to filter and clarify to obtain the roxburghii fermented product.

[0163] The preparation method of the essence comprises the following steps: pre-preparation phase:

[0164] S01 Stir glycerol and p-hydroxyacetophenone in a water bath at 70°C and dissolve completely;

[0165] S02: add deionized water into the container, add butanediol and polyacrylate cross-linked polymer-6, raise the temperature to 80-85°C, keep warm and stir at 800rpm-1000rpm for 10min-20min until uniform; when the material body is cooled to 45-50°C, add the mixture obtained in S01 and stir evenly; continue to cool the material body to 30-35°C, add the sea buckthorn fermentation product, 1,2-hexanediol and disodium EDTA in sequence, and stir evenly.

[0166] Performance Testing

[0167] Test 1: The essences prepared in Examples 1 to 7 were placed at 0°C, 25°C and 40°C for 90 days to observe the properties of the emulsions; the specific results are shown in Table 1.

[0168] Test 2: The essences prepared in Examples 1 to 7 were placed at 25° C. for 12 h, and then at 45° C. for 12 h, respectively. This cycle was repeated 5 times to check the product properties. The specific results are shown in Table 1.

[0169] Table 1 Test results of performance tests 1 and 2

[0170]

[0171] Test 3: Hyaluronidase activity inhibition test (soothing)

[0172] Hyaluronidase is a hyaluronic acid hydrolase. Hyaluronic acid is an acidic mucopolysaccharide widely present in the human body, animals and plants. It is an important component of the extracellular matrix in the human body and is more present in connective tissue, skin, etc. Adding hyaluronidase inhibitors to cosmetics can increase the relative level of hyaluronic acid, keep the skin moisturized, and have an anti-allergic effect. This experiment uses a double antibody sandwich method to detect the inhibition rate of samples on hyaluronidase. The inhibitory factors that may exist in the sample inhibit the activity of hyaluronidase. The still active hyaluronidase binds to the biotin-labeled antibody and then to streptavidin-HRP to form an immune complex, incubate, wash, and then add the color developing solution to develop color. After the reaction is terminated, the absorbance value is tested by an enzyme marker. At the same time, a control group and a blank group are made. By comparing and analyzing the test data, the inhibitory effect of the sample on hyaluronidase is evaluated. From this data, the soothing effect of cosmetics can be relatively reasonably evaluated.

[0173] Prepare Examples 1 to 7, a control group, and a blank group, add samples and hyaluronidase to the corresponding wells of the sample group and the control group, and incubate at 37°C for 10 minutes; then add biotin-labeled antibodies and enzyme-labeled reagents to the corresponding wells of the test group and the control group, incubate at 37°C for 60 minutes, wash the plate 5 times, pat dry, add color developing solutions A and B to all wells, develop at 37°C for 15 minutes, and add stop solution; within 10 minutes, test the OD value at 450nm with an enzyme reader, and calculate the hyaluronidase activity inhibition rate.

[0174] Sample inhibition rate = (average absorbance of the control group - average absorbance of the sample group) / (average absorbance of the control group - average absorbance of the blank group) * 100%. The test results are shown in Table 2.

[0175] Table 2 Test results

[0176] Example Inhibition rate / % Example 1 65.2％ Example 2 59.3％ Example 3 62.4％ Example 4 60.5％ Example 5 57.2％ Example 6 56.8％ Example 7 55.1％

[0177] Test 4: Advanced Glycation End Products AGEs Inhibition Test

[0178] Glycosylated proteins (Advanced Glyeation End Products, AGEs) are the products of Maillard reaction, fat oxidation and other reactions. In this experiment, a Maillard simulation reaction system of bovine serum albumin and glucose was established. The fluorescence intensity was measured at a specific wavelength. The fluorescence intensity was used to represent the AGEs content, and the inhibitory effect of the sample on the formation of glycosylated proteins was calculated.

[0179] Positive control (PC): Aminoguanidine sulfate

[0180] Accurately weigh 0.3 g of aminoguanidine sulfate, dissolve it in 1 mL of PB buffer to prepare a concentration of 300 mg / mL, and dilute it to the required concentration.

[0181] Negative control (NC): PB buffer solution

[0182] Weigh 5.8706 g NaH2PO4·2H2O (sodium dihydrogen) and 0.5646 g Na2HPO4·12H2O (sodium disodium hydrogen), add 200 mL ultrapure water and mix well to obtain phosphate buffer (100 mM) with a pH value of 7.0 (0.5 deviation), which can be adjusted with HCL or NaOH solution.

[0183] Experimental Materials

[0184] Main instruments: fluorescence spectrophotometer, water bath, 96-well plate (black)

[0185] Incubation conditions: 50±1℃

[0186] Main reagents

[0187] Bovine serum albumin, purchased from Sigma, Lot: WXBD5852V;

[0188] D-glucose was purchased from Shanghai Yuanye Biotechnology, Lot: S22J12H137237.

[0189] Experimental procedures

[0190] 5. Prepare AGEs solution: dissolve BSA (50 mg / mL) and D-Glu (0.1 mol / L) in PB, mix well and incubate (50°C, 168 hours).

[0191] Reaction: Add different concentrations of the test solution to the AGEs solution, and set up positive and negative controls and incubate at 50°C for 3 hours.

[0192] Fluorescence value determination: Take 200uL into a 96-well plate and measure the fluorescence value at hex / hem=360nm / 446nm.

[0193] Result analysis: The inhibition rate was calculated according to the following formula

[0194] AGEs inhibition rate (%) = [1-(AB) / (CD)] x 100

[0195] Where: A-fluorescence intensity of AGEs solution with test substance or positive substance added

[0196] B-fluorescence intensity of PB solution with test substance or positive substance added;

[0197] C-fluorescence intensity of AGEs solution without test substance or positive substance;

[0198] D- Fluorescence intensity of PB solution without test substance or positive substance.

[0199] The test results are shown in Table 3.

[0200] Table 3 Test results of test 4

[0201]

[0202]

[0203] Test 5: Elastase test

[0204] The anti-wrinkle and firming effects are mainly characterized by evaluating the inhibition rate of test samples on elastase. Elastase is mainly synthesized and secreted by fibroblasts, and can degrade elastin in the skin, leading to skin aging. The experimental principle of inhibiting elastase is that porcine pancreatic elastase catalyzes the reaction with the enzyme substrate. After adding active substances, the absorbance changes, and the size of the absorbance change reflects the inhibition rate of the elastase inhibitor.

[0205] Reagents

[0206] Elastase (Porcine Pancreas), BR

[0207] N-Succinyl L-Alanyl I-Alanyl L-Alanine, 98% (EGCG), 98% Epigallocatechin Gallate

[0208] Handling of control and test samples

[0209] Essence: Dilute with pure water to a sample concentration of 0.1%:

[0210] Positive control (EGCG): Dilute with water to a positive control concentration of 0.1%:

[0211] Negative control: pure water.

[0212] Test operation steps

[0213] Set up sample group, sample background group, solvent group and solvent background group. Each group needs to set up 3 parallels. Add different reagent solutions to the 96-well plate, shake gently, incubate at 25℃ for 15 minutes, place it in the microplate reader, and measure the absorbance at 410nm.

[0214] Elastase inhibition rate %=(1-(CD) / (AB))*100.

[0215] Wherein: A- is the absorbance of the reaction solution without sample; B- is the absorbance of the reaction solution without sample and enzyme; C- is the absorbance of the reaction solution containing sample and enzyme; D- is the absorbance of the reaction solution containing sample and no enzyme.

[0216] Table 4 Test results of test 5

[0217] Example Inhibition rate / % Example 1 23.425±1.473 Example 2 20.125±0.834 Example 3 21.642±1.123 Example 4 20.946±3.215 Example 5 18.528±2.674 Example 6 17.381±2.152 Example 7 15.186±1.742 Positive control group 82.353±2.145 Negative control group -7.059±3.075

[0218] Test 6: Select eligible subjects according to the subject inclusion and exclusion criteria, and select eligible subjects to ensure that the final number of valid cases is no less than 30.

[0219] ①Skin moisture loss tester TM 300 (Courage+Khazaka, Germany)

[0220] ②Facial Image Analyzer VISIA CR (Canfield, USA)

[0221] ③ Electronic balance JJ3000B (G&G, USA)

[0222] Environmental requirements: Temperature: 21.0℃±1.0℃; Humidity: 50%±10%

[0223] Test process

[0224] ① Recruit volunteers according to the requirements and sign a written informed consent. After washing their faces with water, the subjects sit quietly in a constant temperature and humidity room for 30 minutes. Before enrollment, ask the subjects a series of questions about their medical history, health status, etc. according to the inclusion and exclusion criteria. Screen the eligible subjects for enrollment and ensure that at least 30 people are included.

[0225] ②W0(T0)-basic value: qualified subjects enter the test, and the test order is as follows: doctor's evaluation (skin reaction grading) → lactic acid sting test → VISIA CR (front) → Tewameter (cheek).

[0226] ③ After the test is completed, the subjects will be given instructions on how to use the product. After listening to the instructions, the products and product usage logs will be distributed, and the subjects will use the products at home.

[0227] ④W1: The subjects used the product at home for 1 week, and were followed up on site. After cleansing their face with water, they sat quietly in a constant temperature and humidity chamber for 30 minutes, followed by testing in the following order: doctor's evaluation (skin reaction grading) → lactic acid sting test → VISIA CR (front) → Tewameter (cheek). After completing the test, they filled out a self-assessment questionnaire.

[0228] ⑤ The clinical unit will follow up by phone once a week to check whether the subjects have any skin reactions, and confirm the amount of product used during each follow-up visit.

[0229] Observation and test items

[0230] ① Lactic acid stinging test: Choose the coating method for testing. Apply 50 μL of 10% lactic acid or water to the nasolabial groove on either side at room temperature. The subject's sensitive reaction is evaluated on a 4-point scale at 2.5 minutes and 5 minutes respectively. Then add the two scores together. Those with a (lactic acid-water) score of 3 or more are considered sensitive skin. Parameter explanation: 0 means no stinging; 1 means mild stinging; 2 means moderate stinging; 3 means severe stinging

[0231] Result determination: After using the product, the lactic acid stinging score in the test area was significantly reduced, indicating that the test product has the effect of soothing sensitivity and improving skin tolerance.

[0232] ② Transepidermal water loss rate test: Use Tewameter to test 3 times and take the average value. Parameter explanation: The smaller the test value, the better the skin barrier. Result judgment: After using the product, the transepidermal water loss rate of the test area is significantly reduced, indicating that the tested product has the effect of repairing the skin barrier.

[0233] ③VISIA CR: Use VISIA CR to take frontal photos and use IPP analysis to get the red zone a* value. Parameter explanation: The smaller the a* value, the lower the redness.

[0234] Result determination: After using the product, the a* value of the red zone in the test area was significantly reduced, indicating that the test product has the effect of soothing redness.

[0235] ④Clinical evaluation: Dermatologists will score the skin condition or reaction of the subjects at the application site. The scoring criteria are as follows:

[0236] Skin reaction grading standard for human trial test: no reaction - 0; weak erythema - 1; erythema, infiltration, papules - 2; erythema, edema, papules, blisters - 3; erythema, blisters, bullae - 4; Number of people: 35; Mean age ± SD: 45.43 ± 7.15 years; Minimum age: 31 years; Maximum age: 55 years

[0237] Table 5: Test results of using the product of Example 1 for one week (number of testers: 35 people)

[0238]

[0239]

[0240] Results of the questionnaire analysis on the satisfaction of the subjects

[0241] Table 6 Results of the self-assessment questionnaire on satisfaction of the subjects using the product of Example 1 for one week (number of testers: 35 people)

[0242]

[0243]

[0244] The present invention finds that the roxburghii extract is easy to change color in an aerobic environment during the preparation of the roxburghii fermented product; and the flavonoid components in the roxburghii itself can effectively slow down the occurrence of the above phenomenon. However, due to the difference between the flavonoid components and the vitamin C components, the conventional extraction method or fermentation method cannot achieve the simultaneous analysis of the two, resulting in the separation of the two during the extraction process; in this regard, the present invention performs aerobic and anaerobic fermentation of the roxburghii fermentation process, combines frozen storage, controls the air volume of the sealed fermentation, and the selection of the strains can achieve better fermentation extraction.

[0245] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing a roxburghii fermented product, characterized in that the steps include: S01 Take fresh roxburghii fruit, wash it with clean water and dry it, add solution to crush it and homogenize it, and pour it into a sealed container; S02 then adds culture solution A and strain A to carry out fermentation; after the fermentation is completed, the fermentation product is filtered and frozen; S03: naturally thaw the fermented product obtained in step S02 at room temperature, add culture medium B to continue fermentation, filter to obtain the roxburghii fermented product, and store in a refrigerator.

2. The method for preparing the roxburghii fermented product according to claim 1, characterized in that: The solution in step S01 is a fructose aqueous solution.

3. The method for preparing the roxburghii fermented product according to claim 1, characterized in that: The component of the culture solution A in the S02 is 1000 mL of malt juice.

4. The method for preparing the roxburghii fermented product according to claim 1, characterized in that: The bacterial species A in S02 are yeast and bifidobacterium.

5. The method for preparing the roxburghii fermented product according to claim 1, characterized in that: The components of the culture medium B in the S03 are 10.0 g beef extract powder, 5.0 g peptone, 3.0 g yeast extract powder, 5.0 g glucose, 1.0 g soluble starch, 5.0 g NaCl, 3.0 g NaAc, 0.5 g L-cysteine ​​hydrochloride, and 1000.0 mL distilled water.

6. An essence, characterized in that: The raw materials for preparing the method include: the roxburghii fermented product according to any one of claims 1 to 5.

7. The essence according to claim 6, characterized in that: The essence comprises: deionized water, butylene glycol, glycerin, polyacrylate cross-linked polymer-6, the roxburghii fermented product according to any one of claims 1 to 5, p-hydroxyacetophenone, 1,2-hexanediol, and disodium EDTA.

8. The method for preparing an essence according to claim 6, characterized in that: Here are the steps: S01 Stir glycerol and p-hydroxyacetophenone in a water bath at 70°C and dissolve completely; S02: add deionized water into the container, add butanediol and polyacrylate cross-linked polymer-6, raise the temperature to 80-85°C, keep warm and stir at 800rpm-1000rpm for 10min-20min until uniform; when the material body is cooled to 45-50°C, add the mixture obtained in S01 and stir evenly; continue to cool the material body to 30-35°C, add the sea buckthorn fermentation product, 1,2-hexanediol and disodium EDTA in sequence, and stir evenly.

9. The roxburghii fermented product according to any one of claims 1 to 5, characterized in that: The roxburghii fermented product is applied in the field of skin care.

10. The roxburghii fermented product according to any one of claims 1 to 5, characterized in that: The roxburghii fermented product is used in the fields of moisturizing and anti-aging.

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