Preparation method and application of a Rosa roxburghii Tratt fermentation product

Abstract

The present invention relates to the field of daily chemical technology, and in particular to a preparation method and application of a fermented Rosa roxburghii Tratt product. A preparation method of a fermented Rosa roxburghii Tratt product, the steps comprising: S01 taking fresh Rosa roxburghii Tratt fruits, cleaning them with clean water and then drying them, adding a solution to crush and homogenize them, and pouring them into a sealed container; S02 then adding culture medium A and strain A for fermentation; after fermentation is completed, filtering the fermented product; freezing; S03 naturally thawing the fermented product obtained in step S02 at room temperature, adding culture medium B to continue fermentation, filtering to obtain the fermented Rosa roxburghii Tratt product, and storing it refrigerated. The fermented Rosa roxburghii Tratt product provided by the present invention, and related products have good effects such as whitening, anti-wrinkle, soothing, anti-aging, etc., and the active ingredients are from natural green sources and have high safety.

Description

Technical Field

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

[0002] In recent years, people have become increasingly demanding about the efficacy of cosmetics. Traditional cosmetic ingredients often come with side effects, such as chemical residues and allergic reactions, which have gradually caused concern and attention among consumers. Rosa roxburghii fruit is rich in vitamin C, superoxide dismutase (SOD), organic acids, polysaccharides, flavonoids, polyphenols, and various trace elements, offering antioxidant, anti-aging, whitening, and antibacterial benefits. Rosa roxburghii is a plant that is both medicinal and edible, and its functional ingredients are natural, mild, and non-irritating. Compared to conventional extraction, fermentation technology can break down large active substances into smaller molecules that are more easily absorbed by the skin, thereby enhancing the efficacy of Rosa roxburghii in cosmetics.

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

[0004] CN117070586A is a roxburghii composite polypeptide 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. The roxburghii fruit is used as a fermentation base and bifidobacteria and lactic acid bacteria are used as fermentation sources through cultivation, inactivation and decomposition to obtain a roxburghii composite polypeptide formed by metabolites, cell lipid micromolecule fragments, cell wall components and polysaccharide complexes, i.e., a bifid yeast fermentation product lysate. The fermentation filtrate and filter residue produce a large number of fermentation clusters such as peptide polypeptide groups, bifid yeast, amino acid yeast peptide groups, superoxide dismutase, trace element clusters, mineral enzyme clusters, vitamin C high content groups, and other small molecule clusters, producing a wider range of small molecule biological activities, which can be used for skin repair, nourishing skin, slowing down cell aging, and enhancing skin quality antibodies. It has a great promotion prospect in the fields of cosmetics and food.

[0005] The above patents do not solve the problem that the roxburghii fruit is easy to change color, and the antioxidant properties of the roxburghii fruit after the color change are also greatly reduced. In view of this, the present invention provides a preparation method of a roxburghii fruit fermentation product and its application. 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 in the air, add solution to crush it into a homogenate, and pour it into a sealed container;

[0008] Subsequently, S02 adds culture medium A and strain A for fermentation; after the fermentation ends, the fermented product is filtered; frozen;

[0009] S03 allows the fermented product obtained in step S02 to naturally melt at room temperature, adds culture medium B and continues fermentation, filters to obtain the Rosa roxburghii Tratt fermented product, and stores it refrigerated.

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

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

[0012] As an implementation manner of the present invention, strain A in S02 is yeast and bifidobacterium.

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

[0014] The second aspect of the present invention provides a serum, and its preparation raw materials include the Rosa roxburghii Tratt fermented product described above.

[0015] As an implementation manner of the present invention, the components of the serum include: deionized water, butanediol, glycerol, polyacrylate cross-linked polymer-6, the Rosa roxburghii Tratt 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 preparation method of the serum described above, and the steps are as follows:

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

[0018] S02 adds deionized water to a container, adds butanediol and polyacrylate cross-linked polymer-6, raises the temperature to 80 - 85 °C, keeps warm and stirs at 800 rpm - 1000 rpm for 10 min - 20 min until uniform; when the material cools down to 45 - 50 °C, adds the mixture obtained in S01 and stirs evenly; the material continues to cool down to 30 - 35 °C, and successively adds the Rosa roxburghii Tratt fermented product, 1,2-hexanediol, and disodium EDTA, and stirs evenly.

[0019] The fourth aspect of the invention provides the Rosa roxburghii Tratt fermented product described above, which is characterized in that the Rosa roxburghii Tratt fermented product is applied to the skin care field.

[0020] The fifth aspect of the invention provides the described Rosa roxburghii Tratt ferment, which is applied to the fields of moisturizing and anti-aging.

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

[0022] The Rosa roxburghii Tratt ferment provided by the present invention, and related products have good effects such as whitening, anti-wrinkle, soothing, anti-aging, etc. The active ingredients are from natural green sources and have high safety. Description of the Drawings

[0023] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art. Obviously, the drawings in the following description are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0024] Figure 1 Photo of the essence in Example 1 Detailed Embodiments

[0025] The following will clearly and completely describe the technical solutions of the present invention. Obviously, the described embodiments are some embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope of protection of the present invention.

[0026] The present invention provides a preparation method of a Rosa roxburghii Tratt ferment, and the steps include:

[0027] S01 Take fresh Rosa roxburghii Tratt fruits, wash them clean with water and dry them, add a solution to crush and homogenize, and then pour them into a sealed container;

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

[0029] S02 Then add culture medium A and strain A for fermentation; after fermentation is completed, filter the ferment; freeze;

[0030] The culture medium A is 1000 ml of malt extract, the mass ratio of the Rosa roxburghii Tratt slurry to the culture medium A is 1:10; the mass ratio of the strain A to the culture medium A is 3:100; the strain A contains Saccharomyces cerevisiae and Bifidobacterium; and the inoculation amount ratio is 1:2.

[0031] The Saccharomyces cerevisiae is Rhodosporidium toruloides, China Center for Industrial Culture Collection Management,

[0032] The bifidobacterium is Bifidobacterium animalis subsp. lactis, China Center for Industrial Culture Collection of Microorganisms,

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

[0034] Filtration is carried out using a 0.1 μm filter membrane for clarification; freezing is carried out using liquid nitrogen for cryogenic freezing.

[0035] S03 Naturally melt the fermented product obtained in step S02 at room temperature, add medium B and continue fermentation, filter to obtain the Rosa roxburghii Tratt fermented product, and store it refrigerated.

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

[0037] The fermentation temperature is 30 °C, the fermentation time is 12 h, and after fermentation, filtration is carried out using a 0.1 μm filter membrane for clarification to obtain the Rosa roxburghii Tratt fermented product.

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

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

[0040] The following further explains and illustrates the present invention in combination with specific embodiments.

[0041] Example 1

[0042] This example provides a serum, and its preparation raw materials include

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

[0044] Among them, the preparation method of the Rosa roxburghii Tratt fermented product in this example is as follows:

[0045] S01 Take fresh Rosa roxburghii Tratt fruits, wash them clean with water and dry them, add a solution to break and homogenize, and then pour them into a sealed container to obtain Rosa roxburghii Tratt slurry;

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

[0047] S02 Subsequently, culture medium A and strain A are added for fermentation; after the fermentation is completed, the fermented product is filtered; frozen;

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

[0049] The yeast is Rhodosporidium toruloides, China Center for Industrial Culture Collection of Microorganisms,

[0050] The bifidobacterium is Bifidobacterium animalis subsp. lactis, China Center for Industrial Culture Collection of Microorganisms,

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

[0052] Filtration is carried out using a 0.1 μm filter membrane to clarify; freezing is carried out using liquid nitrogen cryogenic freezing.

[0053] S03 The fermented product obtained in step S02 is naturally melted at room temperature, and chrysanthemum powder is added to continue fermentation in culture medium B, and the rosa roxburghii fermented product is obtained by filtration and stored refrigerated.

[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 S03 are 10.0 g of beef extract powder, 5.0 g of peptone, 3.0 g of yeast extract powder, 5.0 g of glucose, 1.0 g of soluble starch, 5.0 g of NaCl, 3.0 g of NaAc, 0.5 g of L-cysteine hydrochloride, and 1000.0 mL of distilled water.

[0056] The fermentation temperature is 3 C, and the fermentation time is 12 h. After the fermentation is completed, filtration is carried out using a 0.1 μm filter membrane to clarify to obtain the rosa roxburghii fermented product.

[0057] The preparation method of a kind of essence is as follows: Preparation of the preformed phase:

[0058] S01 Glycerol and p-hydroxyacetophenone are stirred and dissolved completely in a water bath at 70 °C;

[0059] Add deionized water into a container, add butylene glycol and polyacrylate crosspolymer-6, heat up to 80 - 85 °C, keep warm and stir at 800 rpm - 1000 rpm for 10 min - 20 min until uniform; wait for the material to cool down to 45 - 50 °C, add the mixture obtained from S01, and stir evenly; continue to cool the material to 30 - 35 °C, and sequentially add the rosa roxburghii tratt ferment, 1,2 - hexanediol, and disodium EDTA, and stir evenly.

[0060] Example 2

[0061] This example provides a serum, and its preparation raw materials include

[0062] Deionized water 88.35%, butylene glycol 4%, glycerol 3%, polyacrylate crosspolymer-6 0.6%, rosa roxburghii tratt ferment 3%, p - hydroxyacetophenone 0.5%, 1,2 - hexanediol 0.5%, disodium EDTA 0.05%.

[0063] Among them, the preparation method of the rosa roxburghii tratt ferment in this example is as follows:

[0064] S01 Take fresh rosa roxburghii tratt fruits, wash them clean with clear water and dry them, add a solution to crush and homogenize, and then pour them into a sealed container to obtain rosa roxburghii tratt slurry;

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

[0066] S02 Then add culture medium A and strain A for fermentation; after fermentation ends, filter the ferment; freeze;

[0067] The culture medium A is 1000 ml of malt extract, and the mass ratio of the rosa roxburghii tratt slurry to the culture medium A is 1:10; the mass ratio of the strain A to the culture medium A is 3:100; the strain A includes Saccharomyces sp. and Bifidobacterium sp.; and the inoculation amount ratio is 1:2.

[0068] The Saccharomyces sp. is Rhodosporidium toruloides, China Center for Industrial Culture Collection,

[0069] The Bifidobacterium sp. is Bifidobacterium animalis subsp. lactis, China Center for Industrial Culture Collection,

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

[0071] Filtration is carried out by filtering and clarifying with a 0.1um filter membrane; freezing is carried out by cryogenic freezing with liquid nitrogen.

[0072] S03 Naturally melt the fermented product obtained in step S02 at room temperature, add medium B and continue fermentation, filter to obtain the Rosa roxburghii Tratt fermented product, and store it refrigerated.

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

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

[0075] The fermentation temperature is 30°C, the fermentation time is 12h, and after fermentation, filtration and clarification are carried out with a 0.1um filter membrane to obtain the Rosa roxburghii Tratt fermented product.

[0076] The preparation method of a kind of essence is as follows: Pre-prepared phase:

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

[0078] S02 Add deionized water to a container, add butanediol and polyacrylate cross-linked polymer-6, heat up to 80 - 85°C, keep warm and stir at 800rpm - 1000rpm for 10min - 20min until uniform; wait for the material to cool down to 45 - 50°C, add the mixture obtained in S01, and stir evenly; continue to cool the material to 30 - 35°C, and sequentially add the Rosa roxburghii Tratt fermented product, 1,2-hexanediol, and disodium EDTA, and stir evenly.

[0079] Example 3

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

[0081] 88.35% of deionized water, 4% of butanediol, 3% of glycerol, 0.6% of polyacrylate cross-linked polymer-6, 3% of Rosa roxburghii Tratt fermented product, 0.5% of p-hydroxyacetophenone, 0.5% of 1,2-hexanediol, 0.05% of disodium EDTA.

[0082] Among them, the preparation method of the Rosa roxburghii Tratt fermented product in this example is as follows:

[0083] S01 Take fresh Rosa roxburghii Tratt fruits, wash them clean with clear water and dry them, add water to crush and homogenize, and then pour them into a sealed container to obtain Rosa roxburghii Tratt slurry;

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

[0085] Subsequently, S02 adds culture medium A and strain A for fermentation; after the fermentation ends, the fermented product is filtered; frozen;

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

[0087] The yeast is Rhodosporidium toruloides, China Center for Industrial Culture Collection of Microorganisms,

[0088] The bifidobacterium is Bifidobacterium animalis subsp. lactis, China Center for Industrial Culture Collection of Microorganisms,

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

[0090] Filtration is carried out with a 0.1 μm filter membrane to clarify; freezing is carried out with liquid nitrogen cryopreservation.

[0091] S03 Naturally melt the fermented product obtained in step S02 at room temperature, and add chrysanthemum powder to the culture medium B for further fermentation, filter to obtain the rosa roxburghii fermented product, and store it refrigerated.

[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 S03 are 10.0 g of beef extract powder, 5.0 g of peptone, 3.0 g of yeast extract powder, 5.0 g of glucose, 1.0 g of soluble starch, 5.0 g of NaCl, 3.0 g of NaAc, 0.5 g of L-cysteine hydrochloride, and 1000.0 mL of distilled water.

[0094] The fermentation temperature is 30 °C, the fermentation time is 12 h, and after the fermentation ends, filtration is carried out with a 0.1 μm filter membrane to clarify to obtain the rosa roxburghii fermented product.

[0095] The preparation method of a kind of essence is as follows: Preparing phase:

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

[0097] Add deionized water into a container, add butylene glycol and polyacrylate crosspolymer-6, heat up to 80-85 °C, keep warm and stir at 800 rpm - 1000 rpm for 10 min - 20 min until uniform; wait for the material to cool down to 45-50 °C, add the mixture obtained in S01, and stir evenly; continue to cool the material to 30-35 °C, and add the rosa roxburghii tratt ferment, 1,2-hexanediol and disodium EDTA in sequence, and stir evenly.

[0098] Example 4

[0099] This example provides a serum, and its preparation raw materials include

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

[0101] Among them, the preparation method of the rosa roxburghii tratt ferment in this example is as follows:

[0102] S01 Take fresh rosa roxburghii tratt fruits, wash them clean with clear water and dry them, add a solution to crush and homogenize, and then pour them into a sealed container to obtain rosa roxburghii tratt slurry;

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

[0104] S02 Then add culture solution A and strain A for fermentation; after fermentation, filter the ferment;

[0105] The culture solution A is 1000 ml of malt extract, and the mass ratio of the rosa roxburghii tratt 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 includes Rhodosporidium toruloides and Bifidobacterium animalis subsp. lactis; and the inoculation amount ratio is 1:2.

[0106] The Rhodosporidium toruloides is Rhodosporidium toruloides, China Center for Industrial Culture Collection Management,

[0107] The Bifidobacterium animalis subsp. lactis is Bifidobacterium animalis subsp. lactis, China Center for Industrial Culture Collection Management,

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

[0109] Filtration is carried out using a 0.1um filter membrane to clarify.

[0110] S03 Add the fermented product obtained in step S02 and chrysanthemum powder to medium B and continue fermentation. Filter to obtain the Rosa roxburghii Tratt fermented product and store it refrigerated.

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

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

[0113] The fermentation temperature is 30°C, the fermentation time is 12h, and after fermentation, filtration is carried out using a 0.1um filter membrane to clarify to obtain the Rosa roxburghii Tratt fermented product.

[0114] The preparation method of a kind of essence is as follows: Preparing phase:

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

[0116] S02 Add deionized water to a container, add butanediol and polyacrylate cross-linked polymer-6, heat up to 80 - 85°C, keep warm and stir at 800rpm - 1000rpm for 10min - 20min until uniform; wait for the material to cool down to 45 - 50°C, add the mixture obtained in S01, and stir evenly; continue to cool the material to 30 - 35°C, and successively add the Rosa roxburghii Tratt fermented product, 1,2-hexanediol, and disodium EDTA, and stir evenly.

[0117] Example 5

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

[0119] 88.35% deionized water, 4% butanediol, 3% glycerol, 0.6% polyacrylate cross-linked polymer-6, 3% Rosa roxburghii Tratt fermented product, 0.5% p-hydroxyacetophenone, 0.5% 1,2-hexanediol, 0.05% disodium EDTA.

[0120] Among them, the preparation method of the Rosa roxburghii Tratt fermented product in this example is as follows:

[0121] S01 Take fresh Rosa roxburghii Tratt fruits, wash them clean with water and dry them, add the solution to crush and homogenize, and then pour them into a sealed container to obtain Rosa roxburghii Tratt slurry;

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

[0123] Subsequently, S02 adds culture medium A and strain A for fermentation; after fermentation, the fermented product is filtered; and it is stored refrigerated.

[0124] The culture medium A is 1000 ml of wort, and the mass ratio of the roselle slurry to the culture medium A is 1:10; the mass ratio of the strain A to the culture medium A is 3:100; the strain A contains yeast and Bifidobacterium; and the inoculation amount ratio is 1:2.

[0125] The yeast is Rhodosporidium toruloides, China Center for Industrial Culture Collection Management,

[0126] The Bifidobacterium is Bifidobacterium animalis subsp. lactis, China Center for Industrial Culture Collection Management,

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

[0128] Filtration is carried out using a 0.1 μm filter membrane to clarify.

[0129] The mass ratio of the fermented product obtained by 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 a kind of essence is as follows: prefabricated phase:

[0131] S01 Dissolve glycerol and p-hydroxyacetophenone completely by stirring in a water bath at 70 °C;

[0132] S02 Add deionized water into the container, add butanediol and polyacrylate crosslinked polymer-6, heat up to 80 - 85 °C, keep warm and stir at 800 rpm - 1000 rpm for 10 min - 20 min until uniform; wait for the material to cool down to 45 - 50 °C, add the mixture obtained by S01, and stir evenly; continue to cool the material to 30 - 35 °C, and add the roselle fermented product, 1,2-hexanediol, and disodium EDTA in sequence, and stir evenly.

[0133] Example 6

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

[0135] Deionized water 88.35%, butylene glycol 4%, glycerin 3%, polyacrylate crosspolymer - 6 0.6%, Rosa roxburghii Tratt ferment 3%, p - hydroxyacetophenone 0.5%, 1,2 - hexanediol 0.5%, disodium EDTA 0.05%.

[0136] Among them, the preparation method of the Rosa roxburghii Tratt ferment in this example is as follows:

[0137] S01 Take fresh Rosa roxburghii Tratt fruits, wash them clean with clear water and dry them, add the solution to crush and homogenize, and then pour them into a sealed container to obtain Rosa roxburghii Tratt slurry;

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

[0139] S02 Then add culture medium A and strain A for fermentation; after fermentation, filter the ferment and store it refrigerated.

[0140] The culture medium A is 1000 ml of malt extract, the mass ratio of the Rosa roxburghii Tratt slurry to the culture medium A is 1:10; the mass ratio of the strain A to the culture medium A is 3:100; the strain A contains Saccharomyces and Bifidobacterium; and the inoculation amount ratio is 1:2.

[0141] The Saccharomyces is Rhodosporidium toruloides, China Center for Industrial Culture Collection,

[0142] The Bifidobacterium is Bifidobacterium animalis subsp. lactis, China Center for Industrial Culture Collection,

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

[0144] Filtration is carried out using a 0.1um filter membrane to clarify.

[0145] The mass ratio of the ferment 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 a kind of essence is as follows: Pre - formed phase:

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

[0148] Add deionized water into a container, add butylene glycol and polyacrylate crosspolymer-6, heat up to 80-85 °C, keep warm and stir at 800 rpm - 1000 rpm for 10 min - 20 min until uniform; wait for the material to cool down to 45-50 °C, add the mixture obtained in S01, and stir evenly; continue to cool the material to 30-35 °C, and sequentially add the rosa roxburghii tratt ferment, 1,2-hexanediol, and disodium EDTA, and stir evenly.

[0149] Example 7

[0150] This example provides a serum, and its preparation raw materials include

[0151] Deionized water 88.35%, butylene glycol 4%, glycerol 3%, polyacrylate crosspolymer-6 0.6%, rosa roxburghii tratt ferment 3%, p-hydroxyacetophenone 0.5%, 1,2-hexanediol 0.5%, disodium EDTA 0.05%.

[0152] Among them, the preparation method of the rosa roxburghii tratt ferment in this example is as follows:

[0153] S01 Take fresh rosa roxburghii tratt fruits, wash them clean with clear water and dry them, add a solution to crush and homogenize, and then pour them into a sealed container to obtain rosa roxburghii tratt slurry;

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

[0155] S02 Then add culture medium A and strain A for fermentation; after fermentation, filter the ferment; freeze;

[0156] The culture medium A is 1000 ml of malt extract, and the mass ratio of the rosa roxburghii tratt slurry to the culture medium A is 1:10; the mass ratio of the strain A to the culture medium A is 3:100; the strain A is yeast. The yeast is Rhodosporidium toruloides, China Center for Industrial Culture Collection,

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

[0158] Filtration is carried out with a 0.1 μm filter membrane to clarify; freezing is carried out with liquid nitrogen cryogenic freezing.

[0159] S03 Naturally melt the ferment obtained in step S02 at room temperature, add chrysanthemum powder and continue fermentation in culture medium B, filter to obtain rosa roxburghii tratt ferment, and store it refrigerated.

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

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

[0162] The fermentation temperature is 30 °C, the fermentation time is 12 h, and after fermentation, filtration and clarification are carried out using a 0.1-μm filter membrane to obtain the rosa roxburghii tratt fermented product.

[0163] The preparation method of a kind of essence is as follows: Preparation of the preformed phase:

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

[0165] S02 adds deionized water into a container, adds butanediol and polyacrylate cross-linked polymer-6, heats up to 80 - 85 °C, keeps warm and stirs at 800 - 1000 rpm for 10 - 20 min until uniform; when the material cools down to 45 - 50 °C, add the mixture obtained from S01 and stir evenly; the material continues to cool down to 30 - 35 °C, and then add the rosa roxburghii tratt fermented product, 1,2-hexanediol, and disodium EDTA in sequence, and stir evenly.

[0166] Performance test

[0167] Test 1: Place the essence prepared in Examples 1 - 7 at 0 °C, 25 °C, and 40 °C for 90 days to observe the emulsion properties; the specific results are shown in Table 1.

[0168] Test 2: Place the essence prepared in Examples 1 - 7 at 25 °C for 12 h, then place it at 45 °C for 12 h, and repeat this cycle 5 times to check the product properties, and 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 experiment (soothing)

[0172] Hyaluronidase is a hyaluronic acid hydrolase. Hyaluronic acid is an acidic mucopolysaccharide widely present in the bodies of humans, animals, plants, etc. It is an important component of the extracellular matrix in the human body and is present in relatively large amounts in connective tissues, skin, etc. Adding a hyaluronidase inhibitor to cosmetics can increase the relative level of hyaluronic acid, keep the skin moist, and have an anti-allergic effect. In this experiment, the double antibody sandwich method was used to detect the inhibition rate of the sample on hyaluronidase. The inhibitory factor that may exist in the sample inhibits the activity of hyaluronidase. The still-active hyaluronidase binds to the biotin-labeled antibody and then binds to streptavidin-HRP to form an immune complex. After incubation and washing, a chromogenic solution is added for color development. After terminating the reaction, the absorbance value is measured by an enzyme-linked immunosorbent assay (ELISA) reader. At the same time, a control group and a blank group are set up. By comparing and analyzing the experimental 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 1-7, the control group, and the blank group. Add the sample and hyaluronidase to the corresponding wells of the sample group and the control group, and incubate at 37°C for 10 min; then add the biotin-labeled antibody and the enzyme-labeled reagent to the corresponding wells of the experimental group and the control group, incubate at 37°C for 60 min, wash the plate 5 times, pat dry, and then add chromogenic solutions A and B to all wells, develop color at 37°C for 15 min, and add the termination solution; within 10 min, measure the OD value at 450 nm with an ELISA reader and calculate the inhibition rate of hyaluronidase activity.

[0174] Sample inhibition rate = (average absorbance value of the control group - average absorbance value of the sample group) / (average absorbance value of the control group - average absorbance value of the blank group) * 100%, and 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: Inhibition Test of Advanced Glycation End Products (AGEs)

[0178] Advanced Glycation End Products (AGEs) are products generated during the Maillard reaction, lipid oxidation, and other reaction processes. In this experiment, a Maillard simulation reaction system of bovine serum albumin and glucose was established. At a specific wavelength, the fluorescence intensity was measured, and the AGEs content was represented by the fluorescence intensity to calculate the inhibitory effect of the sample on the generation of glycated proteins.

[0179] Positive control (PC): aminoguanidine sulfate

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

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

[0182] Weigh 5.8706 g of NaH2PO4·2H2O (sodium dihydrogen phosphate) and 0.5646 g of Na2HPO4·12H2O (disodium hydrogen phosphate), add 200 mL of ultrapure water and mix well to obtain a 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 °C

[0186] Main reagents

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

[0188] D-Glucose, purchased from Shanghai Yuanye Bio-Technology Co., Ltd., Lot: S22J12H137237.

[0189] Experimental procedures

[0190] 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 solutions to the AGEs solution, and set positive and negative controls and incubate at 50 °C for 3 h.

[0192] Determination of fluorescence value: Take 200 μL each and transfer them to a 96-well plate, and measure the fluorescence value at hex / hem = 360 nm / 446 nm.

[0193] Result analysis: Calculate the inhibition rate according to the following formula

[0194] AGEs inhibition rate (%) = [1 - (A - B) / (C - D)] × 100

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

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

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

[0198] D - Fluorescence intensity of the PB solution without the 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 the test sample 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 undergoes a catalytic reaction with the enzyme substrate, and the absorbance changes after adding the active substance. The inhibition rate of the elastase inhibitor is reflected by the magnitude of the change in absorbance.

[0205] Reagents

[0206] Elastase (porcine pancreas), BR

[0207] N-Succinyl-L-alanyl-L-alanyl-L-alanine, 98% (EGCG), 98% epigallocatechin gallate

[0208] Treatment of control and test samples

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

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

[0211] Negative control: Pure water.

[0212] Test operation steps

[0213] Set up a sample group, a sample background group, a solvent group, and a solvent background group. Each group needs to have 3 parallels. Add different reagent solutions to the 96-well plate, shake gently, incubate at 25 °C for 15 min, then place it in an enzyme-labeled instrument and measure the absorbance at 410 nm.

[0214] Elastase inhibition rate % = (1 - (C - D) / (A - B)) * 100.

[0215] Where: A - is the absorbance of the reaction solution without the sample; B - is the absorbance of the reaction solution without the sample and the enzyme; C - is the absorbance of the reaction solution containing the sample and the enzyme; D - is the absorbance of the reaction solution containing the sample and without the enzyme.

[0216] 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 inclusion and exclusion criteria for subjects. Enroll eligible subjects to ensure that the final number of valid cases is not less than 30 people.

[0219] ① Skin Moisture Loss Tester TM 300 (Courage+Khazaka, Germany)

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

[0221] ③ Electronic Balance JJ3000B (Double Jie G&G, USA)

[0222] Environmental Requirements: Temperature: 21.0°C ± 1.0°C; Humidity: 50% ± 10%

[0223] Test Procedure

[0224] ① Recruit and enroll volunteer subjects as required and sign a written informed consent form. After the subjects wash their faces with clean water, they sit quietly in the thermostatic and humidified chamber 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 and enroll eligible subjects to ensure that at least 30 people are finally completed.

[0225] ② W0 (T0) - Baseline Value: Eligible subjects enter the test. The test sequence is as follows: Doctor's assessment (skin reaction grading) → Lactic acid stinging test → VISIA CR (front) → Tewameter (cheek).

[0226] ③ After the test, give the subjects instructions on product use. After the subjects listen to the instructions, distribute the product and the product use log, and the subjects use the product at home.

[0227] ④ W1: The subjects use the product at home for 1 week. Conduct an on-site follow-up visit. After washing their faces with clean water, the subjects sit quietly in the thermostatic and humidified chamber for 30 minutes, and then conduct the test. The test sequence is as follows: Doctor's assessment (skin reaction grading) → Lactic acid stinging test → VISIA CR (front) → Tewameter (cheek). After the test, fill out a self-assessment questionnaire.

[0228] ⑤ The clinical unit conducts a telephone follow-up with the subjects once a week to check for skin reactions and confirm the usage amount of the product each time during the follow-up visit.

[0229] Observation and Test Items

[0230] ①Lactic acid stinging test: The coating method was selected for testing. At room temperature, 50 μL of 10% lactic acid or water was applied to any one nasolabial fold. The sensitive reactions of the subjects were evaluated by the 4-point method at 2.5 min and 5 min respectively, and then the two scores were added together. Those with a (lactic acid - water) score greater than or equal to 3 were considered to have sensitive skin. Parameter explanation: 0 points means no stinging; 1 point means slight stinging; 2 points means moderate stinging; 3 points means severe stinging

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

[0232] ②Transepidermal water loss rate test: The Tewameter was used to test 3 times and the average value was taken. Parameter explanation: The smaller the test value, the better the skin barrier. Result determination: After the product was used, the transepidermal water loss rate in the test area was significantly reduced, indicating that the tested product had the effect of repairing the skin barrier.

[0233] ③VISIA CR: Frontal photos were taken using VISIA CR, and the a* value of the red area was obtained using IPP analysis. Parameter explanation: The smaller the a* value, the lower the degree of flushing.

[0234] Result determination: After the product was used, the a* value of the red area in the test area was significantly reduced, indicating that the tested product had the effect of soothing flushing.

[0235] ④Clinical evaluation: A dermatologist scored 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; Slight erythema - 1; Erythema, infiltration, papules - 2; Erythema, edema, papules, blisters - 3; Erythema, blisters, bullae - 4; Number of people: 35; Average age ± SD: 45.43 ± 7.15 years old; Minimum age: 31 years old; Maximum age: 55 years old

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

[0238]

[0239]

[0240] Analysis results of the subject satisfaction questionnaire

[0241] Table 6 Self-evaluation questionnaire results of subject satisfaction after using the product of Example 1 for 1 week (number of test people: 35)

[0242]

[0243]

[0244] In the process of preparing the Rosa roxburghii Tratt ferment, it was found that the Rosa roxburghii Tratt extract is prone to discoloration in an aerobic environment; among the components of Rosa roxburghii Tratt itself, flavonoid components can effectively slow down the occurrence of the above phenomenon. However, due to the differences between flavonoid components and vitamin C components, conventional extraction methods or fermentation methods cannot simultaneously achieve the simultaneous extraction of both, resulting in the separation during the extraction of the two; in response to this, the present invention combines aerobic and anaerobic fermentation during the Rosa roxburghii Tratt fermentation process, combined with cryopreservation, and controls the air volume of sealed fermentation and the selection of strains to 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 them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A preparation method of a Rosa roxburghii Tratt ferment, characterized in that the steps Including: S01 Take fresh Rosa roxburghii fruits, wash them clean with clear water and dry them. After adding a solution to crush and homogenize, pour them into a sealed container to obtain Rosa roxburghii slurry. The mass ratio of the Rosa roxburghii fruits to the solution is 1:1.5; the solution is 1wt% fructose aqueous solution. S02 Subsequently, add culture medium A and strain A for fermentation; after fermentation is completed, filter the fermented product; freeze. The culture medium A is 1000 ml of malt extract. The mass ratio of the Rosa roxburghii slurry to the culture medium A is 1:10; the mass ratio of the strain A to the culture medium A is 3:100; the strain A contains yeast and Bifidobacterium; and the inoculation amount ratio is 1:

2. The yeast is Rhodosporidium toruloides, China Center for Industrial Culture Collection, 32489. The Bifidobacterium is Bifidobacterium animalis subsp. lactis, China Center for Industrial Culture Collection of Microorganisms, 21712; The fermentation conditions are sealed fermentation, and the total volume of the fermentation broth is 2 / 3 of the total volume of the sealed container; the fermentation temperature is 30 °C, and the fermentation time is 24 h. Filtration is carried out using a 0.1um filter membrane to clarify; freezing is carried out using liquid nitrogen cryogenic freezing. S03 Naturally melt the fermented product obtained in step S02 at room temperature, and add chrysanthemum powder to continue fermentation in culture medium B, filter to obtain the Rosa roxburghii fermented product, and store it refrigerated. 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. The components of the culture medium B in S03 are 10.0 g of beef extract powder, 5.0 g of peptone, 3.0 g of yeast extract powder, 5.0 g of glucose, 1.0 g of soluble starch, 5.0 g of NaCl, 3.0 g of NaAc, 0.5 g of L-cysteine hydrochloride, and 1000.0 mL of distilled water. The fermentation temperature is 30 °C, the fermentation time is 12 h, and after fermentation is completed, filtration is carried out using a 0.1um filter membrane to clarify to obtain the Rosa roxburghii fermented product.

2. An essence, characterized in that: Its preparation raw materials include: the Rosa roxburghii fermented product described in claim 1.

3. The essence according to claim 2, characterized in that, The components of the essence include: deionized water, butanediol, glycerol, polyacrylate cross-linked polymer- 6, the Rosa roxburghii fermented product described in claim 1, p-hydroxyacetophenone, 1,2-hexanediol, disodium EDTA.

4. The preparation method of an essence according to claim 2, wherein, The steps are as follows: S01 Stir and dissolve glycerol and p-hydroxyacetophenone completely in a water bath at 70 °C. S02 Add deionized water to a container, add butanediol and polyacrylate cross-linked polymer-6, heat up to 80 - 85 °C, keep warm and stir at 800 rpm - 1000 rpm for 10 min - 20 min until uniform; wait for the material to cool down to 45 - 50 °C, add the mixture obtained in S01, and stir evenly; continue to cool the material to 30 - 35 °C, and sequentially add the Rosa roxburghii fermented product, 1,2-hexanediol, and disodium EDTA, and stir evenly.

5. The Rosa roxburghii Tratt ferment according to claim 1, wherein The Rosa roxburghii fermented product is applied to the skin care field.

6. The Rosa roxburghii Tratt ferment according to claim 1, characterized in that The Rosa roxburghii fermented product is applied to the fields of moisturizing and anti-aging.

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