Method for preparing skin moisturizer through bidirectional fermentation of schizophyllum commune, skin moisturizer and mask

Abstract

The invention relates to a method for preparing a skin moisturizer through bidirectional fermentation of schizophyllum commune, the skin moisturizer and a mask, and belongs to the technical field of skin care. When the skincare lotion is prepared, the schizophyllum commune seed solution and the lotus root powder are placed in a culture medium to be fermented, and a fermentation product is filtered to obtain the skincare lotion. Schizophyllum commune seed liquid and lotus root powder are added into the culture medium for fermentation, so that the lotus root powder can be utilized to promote growth of schizophyllum commune hyphae to obtain schizophyllum commune fermentation metabolism active products, the schizophyllum commune can be utilized to effectively extract active substances in lotus roots, and by utilizing bidirectional fermentation of the schizophyllum commune and the lotus roots, the fermentation metabolism activity of the schizophyllum commune is improved. The anti-inflammatory performance and the anti-aging performance of the schizophyllum commune fermentation product can be promoted, and the skin moisturizer which is natural in formula, almost free of biological fermentation smell, fresh and cool in skin feeling and good in anti-inflammatory and anti-aging effect can be obtained.

Description

Technical Field

[0001] The present application relates to the technical field of skin care, and in particular to a method for preparing a skin care lotion by bidirectional fermentation using Schizophyllum sp., a skin care lotion, and a facial mask. Background Art

[0002] Currently, the skin care lotions (such as essences, etc.) in commercially available facial masks generally rely on chemically synthesized ingredients (such as preservatives, thickeners) and physically extracted ingredients, and often have disadvantages such as strong irritating odor and sticky feel on the skin. In addition, the current skin care lotions have poor anti-inflammatory and anti-aging effects on the skin. Summary of the Invention

[0003] The present application provides a method for preparing skin care products by bidirectional fermentation using Schizophyllum sp., a skin care lotion and a facial mask. The formula is natural, there is almost no biological fermentation smell, the skin feels refreshing, and there are good anti-inflammatory and anti-aging effects.

[0004] This application is implemented as follows: In a first aspect, an example of the present application provides a method for preparing a skin care lotion using bidirectional fermentation of Schizophyllum sp., comprising: placing Schizophyllum sp. and lotus root powder in a culture medium for fermentation to obtain a fermentation product, and filtering the fermentation product to obtain the skin care lotion.

[0005] In the above implementation process, Schizophyllum is inoculated into a culture medium, and lotus root powder is added to the culture medium. Not only can the lotus root powder be used to promote the growth of Schizophyllum mycelium and obtain the active metabolic products of Schizophyllum fermentation, but the Schizophyllum can also be used to effectively extract the active substances in the lotus root. By utilizing the two-way fermentation of Schizophyllum and lotus root powder, a skin care lotion with a natural formula, almost no biological fermentation odor, a refreshing skin feel, the ability to fight skin inflammation, and relieve skin aging can be obtained.

[0006] In combination with the first aspect, in an optional embodiment of the present application, the method of inoculating Schizophyllum and lotus root powder into a culture medium for fermentation includes: inoculating Schizophyllum seed liquid into the culture medium and fermenting for 1 to 2 days, and then inoculating lotus root powder into the culture medium and fermenting.

[0007] Optionally, the fermentation temperature is 25-30°C.

[0008] In the above implementation process, the seed liquid of Schizophyllum is first inoculated into the culture medium and fermented for 1 to 2 days before the lotus root powder is added. The lotus root matrix is ​​fermented with Schizophyllum as the starting strain. This not only can effectively extract active substances such as vitamins, polysaccharides, trace elements and the like in the lotus root, so that the anti-inflammatory and aging properties of Schizophyllum can be brought into play to a greater extent when applied to the skin, but also can reduce the fermentation odor of the skin care lotion, enhance the refreshing feeling of the skin care lotion, and enhance the overall comfort of the skin care lotion.

[0009] In combination with the first aspect, in an optional embodiment of the present application, the method for preparing the Schizophyllum seed liquid includes: The Schizophyllum strains are activated by using a slant culture medium and placed in an incubator at 27-29° C. for 5-7 days until mycelium grows all over the slant; the slant strains are transferred to a liquid culture medium and expanded and cultured at 27-29° C. and 150-180 r / min for 3-4 days to obtain the activated Schizophyllum seed liquid.

[0010] In the above implementation process, the Schizophyllum strain is inoculated into a slant culture medium at a temperature of 27~29°C and activated and cultured for 5~7 days, and then transferred to a liquid culture medium for expanded culture for 3~4 days. This can promote the proliferation of Schizophyllum hyphae, improve the activity of Schizophyllum, improve the efficiency of subsequent two-way fermentation, improve the stability of metabolically active ingredients in the fermentation product, and improve the anti-inflammatory and anti-aging effects of the skin care lotion.

[0011] In combination with the first aspect, in an optional embodiment of the present application, the inoculation amount of Schizophyllum is 5% to 15%, and the addition amount of lotus root powder is 4% to 8%.

[0012] In the above implementation process, when conducting bidirectional fermentation, adding 5%~15% of Schizophyllum and 4%~8% of lotus root powder to the culture medium can further enhance the anti-inflammatory effect, anti-aging effect, reduce cytotoxicity and promote cell proliferation of the skin care lotion, thereby improving the overall performance of the skin care lotion.

[0013] In combination with the first aspect, in an optional embodiment of the present application, the inoculation amount of Schizophyllum is 5% to 10%, and the addition amount of lotus root powder is 6% to 8%.

[0014] In the above implementation process, bidirectional fermentation using 5% to 10% of Schizophyllum and 6% to 8% of lotus root powder can reduce the cytotoxicity of the skin care lotion to human epidermal cells and improve the cell proliferation-promoting effect of the skin care lotion.

[0015] In combination with the first aspect, in an optional embodiment of the present application, the inoculation amount of Schizophyllum is 5% to 10%, and the addition amount of lotus root powder is 4% to 6%.

[0016] In the above implementation process, bidirectional fermentation using 5% to 10% of Schizophyllum and 4% to 6% of lotus root powder can further enhance the anti-inflammatory effect of the skin care lotion.

[0017] In combination with the first aspect, in an optional embodiment of the present application, the inoculation amount of Schizophyllum is 10% to 15%, and the addition amount of lotus root powder is 6% to 8%.

[0018] Optionally, the inoculation amount of Schizophyllum sp. is 10%, and the addition amount of lotus root powder is 6%.

[0019] In the above implementation process, bidirectional fermentation using 10% to 15% of Schizophyllum and 6% to 8% of lotus root powder can further enhance the anti-aging effect of the skin care lotion.

[0020] In combination with the first aspect, in an optional embodiment of the present application, the culture medium contains, by mass percentage, 0.04% to 0.06% magnesium sulfate, 0.1% to 0.2% potassium dihydrogen phosphate, 1% to 5% glucose, and the remainder is water.

[0021] Optionally, the filtration method includes: centrifuging the fermentation product to remove the bacteria and lotus root powder.

[0022] In the above implementation process, adding 0.04%~0.06% magnesium sulfate, 0.1%~0.2% potassium dihydrogen phosphate and 1%~5% glucose to the culture medium is conducive to the bidirectional fermentation of Schizophyllum and lotus root powder, thereby improving the anti-inflammatory and anti-aging effects of the skin care lotion.

[0023] After the fermentation is completed, the fermentation product is centrifuged to remove the residual bacteria and lotus root powder in the fermentation product, and further obtain a skin care lotion with a light color and a refreshing skin feel.

[0024] In a second aspect, an embodiment of the present application provides a skin care lotion prepared according to the preparation method provided in the first aspect.

[0025] In the above-mentioned implementation process, according to the preparation method provided in the first aspect, Schizophyllum is inoculated into a culture medium, and lotus root powder is added to the culture medium. Not only can the lotus root powder be used to promote the growth of Schizophyllum mycelium and obtain the active metabolic products of Schizophyllum fermentation, but the Schizophyllum can also be used to effectively extract the active substances in the lotus root. The two-way fermentation of Schizophyllum and lotus root powder helps to fully exert the anti-inflammatory and repair characteristics of the Schizophyllum fermentation product when applied to the skin. The obtained skin care lotion has the characteristics of natural formula, almost no biological fermentation smell, refreshing skin feel, and the ability to fight skin inflammation and delay skin aging.

[0026] In a third aspect, an embodiment of the present application provides a facial mask comprising the skin care lotion provided in the second aspect of the present application.

[0027] In the above-mentioned implementation process, the skin care liquid prepared by the preparation method provided in the first aspect of this application is made into a bio-fermentation facial mask. The skin care liquid in the facial mask has the characteristics of natural formula, almost no bio-fermentation smell, refreshing skin feel, ability to fight skin inflammation, and delay skin aging. It can improve the user's comfort, improve skin inflammation, and delay skin anti-aging. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art.

[0029] Figure 1 This is a flow chart of the preparation process of the skin care lotion provided in the examples of this application. DETAILED DESCRIPTION

[0030] The embodiments of the present application will be described in detail below with reference to the examples, but it will be understood by those skilled in the art that the following examples are merely illustrative of the present application and should not be considered as limiting the scope of the present application. In the examples, if specific conditions are not specified, the conditions are carried out according to conventional conditions or manufacturer recommendations. The reagents or instruments used are not specified by the manufacturer and are conventional products that can be purchased commercially.

[0031] At present, the skin care lotions (such as essences, etc.) in commercially available facial masks generally rely on chemically synthesized ingredients (such as preservatives and thickeners) and physically extracted ingredients, and often have disadvantages such as irritating odor and sticky feel on the skin. In addition, the current skin care lotions have poor anti-inflammatory and anti-aging effects on the skin. It is necessary to provide a skin care product with less irritating odor, refreshing, natural and green formula, and anti-inflammatory and anti-aging functions.

[0032] Based on this, the first aspect of the present invention provides a method for preparing a skin care lotion by bidirectional fermentation using Schizophyllum sp., comprising: placing Schizophyllum sp. and lotus root powder in a culture medium for fermentation to obtain a fermentation product, and filtering the fermentation product to obtain the skin care lotion.

[0033] Inoculating Schizophyllum into a culture medium and adding lotus root powder to the culture medium can not only utilize the lotus root powder to promote the growth of Schizophyllum mycelium and obtain the active metabolic products of Schizophyllum fermentation, but also utilize Schizophyllum to effectively extract the active substances in the lotus root. By utilizing the two-way fermentation of Schizophyllum and lotus root powder, the lotus root powder extract can promote the anti-inflammatory and anti-aging properties of Schizophyllum fermentation products, eliminate excess free radicals and inhibit the activity of related enzymes, and obtain a skin care lotion with a natural formula, almost no biological fermentation odor, a refreshing skin feel, the ability to fight skin inflammation, repair skin regeneration, and delay skin aging.

[0034] Lotus root (scientific name: Nelumbo nucifera) is the rhizome of a perennial herbaceous aquatic plant in the genus Nelumbo, family Nymphaeaceae. Growing horizontally in mud, it is oblong, smooth on the surface, and porous and starchy inside. Lotus root leaves are large, round, green on the front and often purplish on the back, with long petioles. Flowers are solitary at the top of long stalks, with white or pink petals. The fruit is an aggregate fruit that splits open upon maturity, revealing black seeds. Lotus root powder is the dried, crushed lotus root tuber.

[0035] The present application does not limit how the lotus root powder is prepared. In some possible embodiments, the lotus root is dried and then crushed, passed through a 40-mesh sieve, and the sieve residue is taken, packaged, and sterilized to prepare the powder.

[0036] Schizophyllum commune Fr. is a fungus in the family Schizophyllum and genus Schizophyllum. Its fruiting bodies are small, scattered or clustered, imbricate, fan-shaped or reniform, sessile or with short stalks. The lid is densely covered with hairs, ranging from white to light gray. The lid margin is involute, striated, and multivalved, and the gills are narrow and curl outward along the longitudinal fissures of the margin.

[0037] The present application does not limit how to inoculate Schizophyllum into the culture medium. In some embodiments, the activated Schizophyllum seed liquid can be inoculated into the culture medium.

[0038] Furthermore, the present application does not limit how the Schizophyllum seed solution is prepared. In some embodiments, please refer to Figure 1 The preparation method provided in the embodiments of the present application also includes preparing a Schizophyllum seed solution, comprising: activating the Schizophyllum spores using a slant culture medium and placing them in an incubator at 27-29°C for 5-7 days until mycelium has fully grown on the slant. The slant culture is then transferred to a liquid culture medium and expanded for 3-4 days at 27-29°C and 150-180 rpm to obtain an activated Schizophyllum seed solution.

[0039] As an example, when activating the Schizophyllum strain to prepare the Schizophyllum seed liquid, the culture temperature can be one of 27°C, 27.5°C, 28°C, 28.5°C or 29°C or a range between any two of them, and the culture time on the slant culture medium can be one of 5 days, 6 days or 7 days or a range between any two of them.

[0040] As an example, when transferring to liquid culture medium for expansion culture, the stirring speed can be one of 150r / min, 155r / min, 160r / min, 165r / min, 170r / min, 175r / min or 180r / min or a range between any two of them.

[0041] As an example, the slant culture medium is a PDA slant culture medium.

[0042] As an example, the liquid culture medium includes 1 wt% to 5 wt% of glucose, 0.1 wt% to 0.2 wt% of potassium dihydrogen phosphate, 0.04 wt% to 0.06% of magnesium sulfate, and the balance is water.

[0043] In some embodiments, the culture medium for bidirectional fermentation of lotus root powder and Schizophyllum can be the same as the above-mentioned liquid culture medium, including 1wt%~5wt% of glucose, 0.1wt%~0.2wt% of potassium dihydrogen phosphate, 0.04wt%~0.06% of magnesium sulfate, and the balance is water, with a natural pH.

[0044] As an example, the mass percentage of glucose in the culture medium may be in the range of one of 1 wt%, 2 wt%, 3 wt%, 4 wt% or 5 wt%, or any two thereof.

[0045] As an example, the mass percentage of potassium dihydrogen phosphate in the culture medium may be 0.1 wt %, 0.15 wt % or 0.2 wt %, or a range between any two of them.

[0046] As an example, the mass percentage of magnesium sulfate in the culture medium may be in the range of one of 0.04 wt %, 0.05 wt % or 0.06 wt %, or any two thereof.

[0047] In some embodiments, the inoculation amount of Schizophyllum is 5% to 15%, and the addition amount of lotus root powder is 4% to 8%. Inoculating the culture medium with appropriate amounts of Schizophyllum and lotus root powder can further enhance the anti-inflammatory and anti-aging effects of the skin care lotion.

[0048] In this application, the inoculation amount of 5% to 15% means that the content of the inoculated Schizophyllum is 5% to 15% (ratio of volume v to volume v) calculated according to the volume of Schizophyllum liquid / the total volume of the culture medium after inoculation of Schizophyllum liquid and lotus root powder.

[0049] In this application, the addition amount of lotus root powder is 4%~8%, which means: calculated according to the mass of lotus root powder / the total volume of the culture medium after inoculating the schizophyllum liquid and lotus root powder, the content of the added lotus root powder is 4%~8% (the ratio of mass w to volume v).

[0050] As an example, the inoculum size of Schizophyllum can be one of 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% or 15%, or a range between any two of these.

[0051] As an example, the added amount of lotus root powder can be 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.8% or 8%, or a range between any two of them.

[0052] Furthermore, in some embodiments, the inoculation amount of Schizophyllum is 5% to 10%, and the addition amount of lotus root powder is 6% to 8%.

[0053] In some embodiments, the inoculation amount of Schizophyllum sp. is 5% to 10%, and the addition amount of lotus root powder is 4% to 6%.

[0054] In some embodiments, the inoculation amount of Schizophyllum sp. is 10% to 15%, and the added amount of lotus root powder is 6% to 8%.

[0055] In some embodiments, the inoculum amount of Schizophyllum is 10%, and the amount of lotus root powder added is 6%. Adding appropriate amounts of Schizophyllum and lotus root powder to the culture medium and utilizing the bidirectional fermentation of the lotus root powder and Schizophyllum can further comprehensively enhance the anti-aging, anti-inflammatory, and cell proliferation effects of the skin care lotion while ensuring that the skin care lotion has no fermented odor and a refreshing feel.

[0056] Further, in some embodiments, please continue to refer to Figure 1 The method of inoculating Schizophyllum and lotus root powder into a culture medium for fermentation includes: inoculating Schizophyllum seed liquid into the culture medium for fermentation for 1-2 days, and then adding lotus root powder into the culture medium for fermentation. When performing bidirectional fermentation of Schizophyllum and lotus root powder, the Schizophyllum seed liquid is first inoculated into the culture medium for fermentation for 1-2 days to allow the Schizophyllum hyphae to fully grow, and then the lotus root matrix is ​​fermented using Schizophyllum as the starting strain to facilitate the extraction of active substances such as vitamins, polyphenols, polysaccharides, and trace elements in the lotus root. These active substances interact with metabolic active products in the Schizophyllum fermentation product to promote the metabolic active products to exert anti-inflammatory, cell proliferation, and anti-aging effects on human skin.

[0057] Furthermore, the present application does not limit the specific temperature for bidirectional fermentation of Schizophyllum and lotus root powder. In some embodiments, the fermentation temperature may be 25-30°C.

[0058] As an example, the temperature during the bidirectional fermentation of Schizophyllum and lotus root powder may be 25° C., 26° C., 27° C., 28° C., 29° C., or 30° C., or a range between any two thereof.

[0059] As an example, the fermentation temperature when bidirectionally fermenting Schizophyllum and lotus root powder may be 28°C.

[0060] Furthermore, the embodiments of the present application do not limit the specific fermentation time when the bidirectional fermentation of Schizophyllum and lotus root powder is carried out. In some embodiments, the bidirectional fermentation time can be 100 to 150 hours.

[0061] As an example, the fermentation time for bidirectional fermentation of Schizophyllum and lotus root powder may be 100 h, 110 h, 120 h, 130 h, 140 h, or 150 h, or a range between any two of these.

[0062] As an example, the fermentation time when the Schizophyllum and lotus root powder are subjected to bidirectional fermentation can be 120 hours.

[0063] Furthermore, after fermentation is complete, the fermentation product can be filtered to remove impurities such as bacteria and lotus root powder residue. This application is not limited to a specific filtering method. In some embodiments, when filtering the fermentation product, a centrifugal treatment method can be used to remove impurities such as bacteria and lotus root powder residue in the fermentation product.

[0064] Furthermore, the fermentation liquid obtained after the centrifugal treatment may be subjected to membrane treatment to further filter out impurities in the fermentation liquid.

[0065] In a second aspect, the present invention provides a skin care lotion prepared according to the above method. The skin care lotion is obtained by bidirectional fermentation of Schizophyllum and lotus root. It is understood that the skin care lotion contains active metabolic products of Schizophyllum and active ingredients of lotus root.

[0066] As an example, the metabolically active products of Schizophyllum include, but are not limited to, Schizophyllan.

[0067] As an example, the active ingredients of lotus root include but are not limited to vitamins, polyphenols, polysaccharides, trace elements and other active substances in lotus root.

[0068] The skin care lotion provided in the embodiments of the present application, through the mutual cooperation of the metabolic active products of Schizophyllum and the active products of lotus root, can promote the anti-inflammatory, cell proliferation and anti-aging effects of the metabolic active products on human skin, and can also make the skin care lotion have the characteristics of no fermentation odor, refreshing skin feel and light color, thereby improving the comprehensive use effect of the skin care lotion.

[0069] Furthermore, the third aspect of the embodiment of the present application also provides an application of a skin care lotion in the field of skin care.

[0070] As an example, a skin care lotion can be applied to a facial mask. Based on this, the present invention also provides a facial mask containing the skin care lotion provided in the present invention, which is a bio-fermentation facial mask. This bio-fermentation facial mask has a natural formula, is essentially free of bio-fermentation odor, and provides a refreshing feel on the skin. It also has excellent anti-inflammatory, anti-aging, and cell proliferation and repair effects.

[0071] As an example, the skin care lotion can also be applied to skin care products such as skin care lotion / emulsion, essence, etc.

[0072] The skin care lotion and its preparation method of the present application are further described in detail below with reference to the examples.

[0073] Example 1 Example 1 provides a skin care lotion, the preparation method of which is as follows: (1) Preparation of lotus root powder: After the lotus root is dried, it is crushed and passed through a 40-mesh sieve. The sieved material is taken to obtain lotus root powder, which is then packaged and sterilized for later use.

[0074] (2) Activation of Schizophyllum strains: Activate the Schizophyllum strains using a slant culture medium and place them in an incubator at 28°C for 5-7 days until the mycelium covers the slant. Transfer the slant strains to a liquid culture medium and culture at 28°C and 160 rpm for 3 days to obtain a Schizophyllum seed solution. The liquid culture medium consists of 5% glucose, 0.1% potassium dihydrogen phosphate, 0.05% magnesium sulfate, and the balance is water, calculated by mass percentage.

[0075] (3) Preparation of culture medium: The culture medium includes the following components in percentage by mass: 0.05% magnesium sulfate, 0.1% potassium dihydrogen phosphate, 3% glucose, pH (natural), and the balance is water.

[0076] (4) Fermentation: The Schizophyllum seed solution prepared in step (2) is inoculated into the culture medium prepared in step (3) for fermentation, with the inoculum amount of Schizophyllum being 10%. After one day of fermentation, the lotus root powder prepared in step (1) is added to the fermentation system at an addition amount of 6%. After fermentation for 120 hours, the fermentation product is centrifuged to remove impurities such as the bacterial cells and lotus root powder residue, thereby obtaining a skin care lotion.

[0077] Example 2 Example 2 provides a skin care lotion, which differs from Example 1 in that: In step (4), the inoculation amount of Schizophyllum is 5%, and the addition amount of lotus root powder is 6%.

[0078] Example 3 Example 3 provides a skin care lotion, which differs from Example 1 in that: In step (4), the inoculation amount of Schizophyllum is 15%, and the addition amount of lotus root powder is 6%.

[0079] Example 4 Example 4 provides a skin care lotion, which differs from Example 1 in that: In step (4), the inoculation amount of Schizophyllum is 10%, and the addition amount of lotus root powder is 4%.

[0080] Example 5 Example 5 provides a skin care lotion, which differs from Example 1 in that: In step (4), the inoculation amount of Schizophyllum is 5%, and the addition amount of lotus root powder is 4%.

[0081] Example 6 Example 6 provides a skin care lotion, which differs from Example 1 in that: In step (4), the inoculation amount of Schizophyllum is 15%, and the addition amount of lotus root powder is 4%.

[0082] Example 7 Example 7 provides a skin care lotion, which differs from Example 1 in that: In step (4), the inoculation amount of Schizophyllum is 10%, and the addition amount of lotus root powder is 8%.

[0083] Example 8 Example 8 provides a skin care lotion, which differs from Example 1 in that: In step (4), the inoculation amount of Schizophyllum is 5%, and the addition amount of lotus root powder is 8%.

[0084] Example 9 Example 9 provides a skin care lotion, which differs from Example 1 in that: In step (4), the inoculation amount of Schizophyllum is 15%, and the addition amount of lotus root powder is 8%.

[0085] Comparative Example 1 Comparative Example 1 provides a skin care lotion, which differs from Example 1 in that: In step (1), the pomelo peel powder is prepared by drying the pomelo peel, crushing it, and passing it through a 40-mesh sieve. The sieved material is taken to obtain the pomelo peel powder, which is then packaged and sterilized for later use.

[0086] In step (4), the Schizophyllum seed liquid from step (2) is inoculated into the culture medium prepared in step (3) for fermentation, with the inoculum amount of Schizophyllum being 10%. After one day of fermentation, the pomelo peel powder from step (1) is added to the fermentation system at an addition amount of 6%. After fermentation for 120 hours, the fermentation product is centrifuged to remove impurities such as the bacterial cells and pomelo peel powder residue, thereby obtaining a skin care lotion.

[0087] Comparative Example 2 Comparative Example 2 provides a skin care lotion, which differs from Example 1 in that: In step (1), the preparation of Buddleja buddleja powder is as follows: the Buddleja buddleja powder is dried, crushed, and passed through a 40-mesh sieve, and the sieved material is taken to obtain the Buddleja buddleja powder, which is then packaged and sterilized for later use.

[0088] In step (4), the Schizophyllum seed liquid from step (2) is inoculated into the culture medium prepared in step (3) for fermentation, with the inoculum amount of Schizophyllum being 10%. After one day of fermentation, the Buddleja buddhist flower powder from step (1) is added to the fermentation system at an addition amount of 6%. After fermentation for 120 hours, the fermentation product is centrifuged to remove impurities such as the bacterial cells and Buddleja buddhist flower powder residue, thereby obtaining a skin care lotion.

[0089] Comparative Example 3 Comparative Example 3 provides a skin care lotion, which differs from Example 1 in that: In step (1), the yam powder is prepared by drying the yam, crushing it, and passing it through a 40-mesh sieve to obtain the yam powder, which is then packaged and sterilized for later use.

[0090] In step (4), the Schizophyllum seed liquid from step (2) is inoculated into the culture medium prepared in step (3) for fermentation, with the inoculum amount of Schizophyllum being 10%. After one day of fermentation, the Chinese yam powder from step (1) is added to the fermentation system at an addition amount of 6%. After fermentation for 120 hours, the fermentation product is centrifuged to remove impurities such as the bacterial cells and Chinese yam powder residue, thereby obtaining a skin care lotion.

[0091] Comparative Example 4 Comparative Example 4 provides a skin care lotion, which differs from Example 1 in that: Do not perform step (1).

[0092] In step (4), the Schizophyllum seed liquid from step (2) is inoculated into the culture medium prepared in step (3) for fermentation, with the inoculum amount of Schizophyllum being 10%. After one day of fermentation, 6% of fresh aloe vera pulp is added to the fermentation system. After 120 hours of fermentation, the fermentation product is centrifuged to remove impurities such as the bacterial cells and fresh aloe vera pulp residue, thereby obtaining a skin care lotion.

[0093] Some process parameters of Examples 1-9 and Comparative Examples 1-4 are shown in Table 1.

[0094] Table 1 Raw material composition

[0095] Test example (1) Skin feel test Fifteen volunteers rated the fermentation filtrates of Example 1 and Comparative Examples 1-5 based on three indicators: before use (smell), during use (stickiness, absorption speed), and after use (moisturizing, mildness), as well as their overall user experience. The ratings were based on a scale of 0-100, with 0-20 indicating very dissatisfied, 20-40 indicating dissatisfied, 40-60 indicating fair, 60-80 indicating satisfied, and 80-100 indicating very satisfied. The rating results are shown in Table 2.

[0096] Table 2 Summary of skin care lotion skin sensory evaluation scores

[0097] Results Analysis: Table 2 shows that the skin care lotion obtained by bidirectional fermentation of Schizophyllum and lotus root powder in Example 1 received the highest overall user experience rating, with essentially no fermentation odor, high satisfaction with the sticky feeling on the skin, and gentle application. All indicators before, during, and after use were highly satisfactory. Furthermore, the skin care lotion provided in this example also exhibited excellent moisturizing and retention effects and a high skin absorption rate.

[0098] (2) Cytotoxicity test The Cell Viability Kit (CCK-8) can be used for simple and accurate cell proliferation and toxicity analysis. Its basic principle is that the reagent contains WST-8 (chemical name: 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonatophenyl)-2H-tetrazolium monosodium salt), which is reduced by cellular dehydrogenases into a highly water-soluble yellow formazan dye under the action of the electron carrier 1-methoxy-5-methylphenazinium dimethyl sulfate (1-Methoxy PMS). The amount of formazan generated is proportional to the number of viable cells. This property allows direct analysis of cell proliferation and toxicity. The CCK-8 assay is a highly sensitive, non-radioactive colorimetric method for determining the number of viable cells in cell proliferation or toxicity experiments, and is a viable alternative to the traditional MTT assay.

[0099] The specific experimental steps are as follows: 1. Add 100 μL of cells to each well of a 96-well plate (leave two blank groups untreated and add the same volume of culture medium). Incubate the cells in a 37°C, 5% CO2 cell culture incubator for 24 h.

[0100] Note: A. Typically, 100 μL of culture medium added to each well for cell proliferation assays contains approximately 2,000 cells. For cytotoxicity assays, 100 μL of culture medium added to each well contains approximately 5,000 to 10,000 cells. (The specific number of cells used per well depends on cell size, cell proliferation rate, etc.)

[0101] B. The culture temperature depends on the cell type. Generally, 37°C is recommended for mammalian cells. For other species, the appropriate temperature should be selected based on the cell culture conditions.

[0102] 2. Add 10 μL of drug at different concentrations to each well for stimulation (for cells added in step 1, leave two control groups without drug and add the same volume of culture medium).

[0103] 3. Incubate the 96-well plate in a cell culture incubator at 37°C, 5% CO2 in air, and 100% humidity for an appropriate period of time.

[0104] Note: Same as above. Select appropriate incubation conditions and time based on the cell needs of different experiments.

[0105] 4. Add 10 μL of CCK-8 solution to each well and incubate at 37°C in a 5% CO2 incubator for 1–4 h.

[0106] Note: Same as above. Select appropriate incubation conditions and time based on the cell needs of different experiments.

[0107] 5. Measure the absorbance at 450 nm using a microplate reader.

[0108] 6. If you are not measuring the OD (absorbance) value temporarily, add 10 μL of 0.1 M HCl solution or 1% (w / v) SDS solution to each well and store the culture plate covered and protected from light at room temperature. The absorbance will not change within 24 hours.

[0109] 7. Test data processing: A. Cell viability: Subtract the background OD value (blank group) from the OD value of each test well, and calculate the mean ± SD (standard deviation) of the OD values ​​of each replicate well.

[0110] Cell viability %=(OD of drug-treated cells / OD of control cells)×100%.

[0111] The test data is shown in Table 3.

[0112] Table 3 Survival rate of human skin fibroblasts (BJ)

[0113] Analysis of results: It can be seen from Table 3 that in Examples 1 to 9 of the present application, the cell survival rates of the bidirectional fermentation using Schizophyllum and lotus root powder were 81.58% to 117.28%, which were higher than the cell survival rates of 66.20% to 80.33% in Comparative Examples 1 to 5. This indicates that the skin care lotion obtained by bidirectional fermentation using Schizophyllum and lotus root powder in the examples of the present application is basically non-toxic to human skin fibroblasts (BJ), and the skin care lotion obtained by bidirectional fermentation using 10% Schizophyllum and 6% lotus root powder in Example 1 also has a significant effect of promoting cell proliferation and can repair the skin.

[0114] (3) Cell anti-inflammatory and anti-aging tests Inflammation is an adaptive response triggered by harmful stimuli such as ischemic injury, infection, and tissue trauma. Bacterial lipopolysaccharide (LPS) is the primary pathogenic component that triggers inflammatory responses. LPS-induced immune cells, such as macrophages, release large amounts of inflammatory factors such as interleukin-1β (IL-1β), which can react on senescent cells and their neighboring cells, accelerating their aging process. This experiment used bacterial lipopolysaccharide (LPS)-stimulated human skin fibroblasts (BJ) as an inflammatory aging model. The expression levels of the inflammatory factors interleukin-1β (IL-1β), interleukin-18 (IL-18), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), and the anti-aging genes collagen type I and III (COL1 and COL3) were measured before and after sample addition.

[0115] The specific experimental steps are as follows: 1. Cell culture: BJ cells were cultured in a 37°C, 5% CO2 incubator and passaged in the logarithmic phase.

[0116] 2. Cell Grouping and Intervention: Cells in the logarithmic growth phase were plated at a density of 2×10⁵ / mL and divided into a normal group, a model group (control group), and a drug-treated group. All cells except the normal group were treated with lipopolysaccharide (LPS) to establish an inflammatory aging model. The drug-treated groups were pre-treated with test samples at varying concentrations for 1 hour, followed by the addition of LPS (1 μg / mL).

[0117] Analysis of mRNA expression levels of inflammation-related genes: Cells were grouped according to the grouping method in step 2, and the test samples were treated for 24 hours. The culture supernatant was discarded and the cells were washed twice with PBS. According to the kit instructions, RNA was reverse transcribed to generate cDNA (25°C, 10 min; 50°C, 30 min; 85°C, 5 min), and RT-qPCR analysis was performed (95°C, 2 min; 95°C, 15 s; 63°C, 25 s; 72°C, 15 s; amplification for 40 cycles). β-actin was used as an internal reference, and 2 -△△CT Method for analysis.

[0118] The test results of the expression levels of anti-inflammatory genes (NF-κB, IL-1β, IL-18) in various examples and comparative examples are shown in Table 4.

[0119] Table 4 Expression levels of anti-inflammatory genes (NF-κB, IL-1β, IL-18)

[0120] Results Analysis: As can be seen from Table 4, a comparison between the control group and Examples 1-9 shows that the inflammatory aging model constructed in the control group did not contain the skin care lotion, and the expression of anti-inflammatory genes could not be inhibited. The expression levels of the anti-inflammatory genes NF-κB, IL-1β, and IL-18 in the control group were as high as 2.94, 2.02, and 2.75, respectively. However, when the skin care lotion obtained by bidirectional fermentation of Schizophyllum and lotus root powder in Examples 1-9 of this application was added to the inflammatory aging model, the expression levels of the NF-κB anti-inflammatory gene did not exceed 0.84 (lower than 2.94 in the control group), the expression level of the IL-1β anti-inflammatory gene did not exceed 0.89 (lower than 2.02 in the control group), and the expression level of the IL-18 anti-inflammatory gene did not exceed 0.66 (lower than 2.75 in the control group). The expression levels of the anti-inflammatory genes in Examples 1-9 were all lower than those in the control group. The lower the expression level of the anti-inflammatory gene, the better the anti-inflammatory effect.

[0121] Moreover, by comparing Examples 1 to 9, it can be seen that in a culture medium with the same nutrient content, the anti-inflammatory effect varies with the inoculation amount of Schizophyllum and lotus root, and there is a normal distribution. When 10% Schizophyllum and 6% lotus root are inoculated, the anti-inflammatory effect is the best.

[0122] Comparing Example 1 with Comparative Examples 1 to 4, it can be seen that Example 1 and Comparative Examples 1 to 4 all contain 10% of Schizophyllum and 6% of different powders. The skin care liquid obtained by bidirectional fermentation using 10% Schizophyllum and 6% lotus root powder in Example 1 has an expression level of 0.24 for the NF-κB anti-inflammatory gene, an expression level of 0.48 for the IL-1β anti-inflammatory gene, and an expression level of 0.24 for the IL-18 anti-inflammatory gene, which is lower than the expression levels of the skin care liquid obtained by bidirectional fermentation using 10% Schizophyllum with 6% grapefruit peel powder, flos montana powder, yam powder, and fresh aloe vera pulp. This shows that the present application embodiment uses lotus root powder and Schizophyllum for bidirectional fermentation, which can promote the anti-inflammatory function of Schizophyllum. If other materials other than lotus root, such as yam, aloe, grapefruit peel, or flos montana, are used for bidirectional fermentation with Schizophyllum, it cannot effectively promote the anti-inflammatory function of the Schizophyllum fermentation product.

[0123] The test results of the expression levels of anti-inflammatory genes (NF-κB, IL-1β, IL-18) in various examples and comparative examples are shown in Table 5.

[0124] Table 5 Expression levels of anti-aging genes (COL1, COL3)

[0125] Results Analysis: As can be seen from Table 5, comparing the control group and Examples 1-9, the inflammatory aging model constructed in the control group, which did not include the addition of skin care lotion, failed to promote the expression of anti-aging genes. The expression levels of the anti-aging genes COL1 and COL3 were only 0.23 and 0.14, respectively. However, when the skin care lotion obtained by bidirectional fermentation of Schizophyllum and lotus root powder in Examples 1-9 of this application was added to the inflammatory aging model, the expression levels of the COL1 anti-aging gene were no less than 1.55 (higher than 0.23 in the control group) and no less than 2.27 (higher than 0.14 in the control group). The expression levels of the anti-aging genes in Examples 1-9 were all higher than those in the control group. Higher expression levels of anti-aging genes indicate better anti-aging effects.

[0126] Comparing Example 1 with Comparative Examples 1 to 4, it can be seen that Example 1 and Comparative Examples 1 to 4 both contain 10% of Schizophyllum and 6% of different powders. In Example 1, the skin care liquid obtained by bidirectional fermentation using 10% of Schizophyllum and 6% of lotus root powder has an expression level of 3.94 for the COL1 anti-aging gene and an expression level of 5.63 for the COL3 anti-aging gene, which are higher than the expression levels of the skin care liquids obtained by bidirectional fermentation using 10% of Schizophyllum with 6% of pomelo peel powder, flos montana powder, yam powder, and fresh aloe vera pulp, respectively. This indicates that the bidirectional fermentation using lotus root powder and Schizophyllum in the present application can promote the anti-aging function of Schizophyllum. However, if other materials other than lotus root, such as yam, aloe vera, pomelo peel, or flos montana, are used for bidirectional fermentation with Schizophyllum, the anti-aging function of the Schizophyllum fermentation product cannot be effectively promoted.

[0127] In summary, the embodiment of the present application uses Schizophyllum and lotus root powder for bidirectional fermentation to obtain a skin care lotion with a natural formula, almost no biological fermentation odor, a refreshing skin feel, the ability to fight skin inflammation, promote cell proliferation and repair the skin, and delay skin aging.

[0128] The foregoing description is merely a preferred embodiment of the present application and is not intended to limit the present application. Persons skilled in the art will readily appreciate that various modifications and variations are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present application shall be included within the scope of protection of the present application.

Claims

1. A method for preparing a skin care lotion by bidirectional fermentation using Schizophyllum sp., characterized in that: include: placing Schizophyllum and lotus root powder in a culture medium for fermentation to obtain a fermentation product; The fermentation product is filtered to obtain the skin care lotion.

2. The preparation method according to claim 1, characterized in that The method of inoculating Schizophyllum and lotus root powder into a culture medium for fermentation comprises: inoculating Schizophyllum seed liquid into the culture medium for fermentation for 1 to 2 days, then adding the lotus root powder into the culture medium for fermentation; Optionally, the fermentation temperature is 25-30° C., and the fermentation time is 100-150 hours.

3. The preparation method according to claim 2, characterized in that The preparation method of the Schizophyllum seed liquid comprises: The Schizophyllum strains are activated by using a slant culture medium and placed in an incubator at 27-29° C. for 5-7 days until mycelium grows all over the slant; the slant strains are transferred to a liquid culture medium and expanded and cultured at 27-29° C. and 150-180 r / min for 3-4 days to obtain the activated Schizophyllum seed liquid.

4. The preparation method according to claim 1, characterized in that The inoculation amount of the Schizophyllum is 5% to 15%, and the addition amount of the lotus root powder is 4wt% to 8wt%.

5. The preparation method according to claim 4, characterized in that The inoculation amount of the Schizophyllum is 5% to 10%, and the addition amount of the lotus root powder is 6% to 8%.

6. The preparation method according to claim 4, characterized in that The inoculation amount of the Schizophyllum is 5% to 10%, and the addition amount of the lotus root powder is 4% to 6%.

7. The preparation method according to claim 4, characterized in that The inoculation amount of the Schizophyllum is 10% to 15%, and the addition amount of the lotus root powder is 6% to 8%; Optionally, the inoculation amount of the Schizophyllum sp. is 10%, and the addition amount of the lotus root powder is 6%.

8. The preparation method according to any one of claims 1 to 7, characterized in that The culture medium contains, by weight percentage, 0.04% to 0.06% magnesium sulfate, 0.1% to 0.2% potassium dihydrogen phosphate, 1% to 5% glucose, and the balance is water; Optionally, the filtration method includes: centrifuging the fermentation product to remove bacteria and the lotus root powder.

9. A skin care lotion prepared according to the preparation method according to any one of claims 1 to 8.

10. A facial mask, characterized in that: The skin care lotion according to claim 9.

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