Preservative-free white truffle fermented skin tendering mask and preparation method thereof

Abstract

The invention relates to a preservative-free white truffle fermented skin tendering mask and a preparation method of the preservative-free white truffle fermented skin tendering mask. Comprising 1-5 parts of a white truffle fermentation filtrate, 0.1-2 parts of a bamboo leaf extract, 0.1-1 part of a green tea extract, 1-2 parts of auricularia auricula polysaccharide, 1-1.5 parts of a tremella fuciformis extract, 1-3 parts of a bacillus fermentation product, 0.5-1.5 parts of a wheat germ extract, 1-2 parts of a dendrobium officinale stem extract, 1-5 parts of glycerin and 70-90 parts of water. The white truffle is subjected to enzymolysis through papain, then fermentation is conducted through saccharomycetes, white truffle fermentation filtrate with high water-soluble protein content is obtained, bamboo leaf extract, green tea extract and black fungus polysaccharide are added into the fermentation filtrate, and the green and preservative-free white truffle fermented skin tendering mask is prepared.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to a preservative-free white truffle fermented skin rejuvenation mask and a preparation method thereof. Background Art

[0002] Facial mask is a kind of beauty care product. After years of development, it has become a must-have item for women's skin care. The main function of facial mask is to deliver nutrients into the skin through the film material, so as to achieve the effect of skin care. However, the moisture and nutrients in the facial mask are easily contaminated by microorganisms such as bacteria and mold, resulting in deterioration. If preservatives are not added, the facial mask is easily infected by microorganisms, produces odor, corruption, and even seriously affects the user's skin health. Long-term use of preservatives has potential harm to the skin, including allergies, irritation, dryness and damage to the skin barrier.

[0003] White truffle is a truffle mushroom that contains crude protein, fat, amino acids, polyphenols, polysaccharides, multiple minerals, trace elements and carbohydrates. It has always been known as the "white diamond" and has extremely high nutritional value. White truffle is not only a top-grade food, but also has high medicinal value. It is known as the "king of immunity" together with Ganoderma lucidum. White truffle also has a variety of biologically active ingredients, which can not only inhibit the production of melanin, thereby achieving the effect of whitening and brightening the skin; it also has antioxidant and anti-aging activities, which can revitalize cells.

[0004] The invention patent with publication number CN114748381A discloses a method for preparing white truffle fermentation liquid, using white truffle extract as raw material and fermenting with Bacillus. The white truffle extract obtained by water extraction improves the bioavailability of white truffle. In order to improve the bioavailability of white truffle while improving the safety of white truffle use, it is urgent to develop a preservative-free white truffle fermentation skin rejuvenation mask and its preparation method. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides a preservative-free white truffle fermented skin-rejuvenating mask and a preparation method thereof. White truffles are enzymatically hydrolyzed by papain and then fermented by yeast to obtain a white truffle fermentation filtrate with a high water-soluble protein content. Bamboo leaf extract, green tea extract and black fungus polysaccharide are added to the fermentation filtrate to prepare a preservative-free white truffle fermented skin-rejuvenating mask.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A preservative-free white truffle fermented skin rejuvenation mask comprises, by weight: white truffle fermentation filtrate, bamboo leaf extract, green tea extract, and black fungus polysaccharide.

[0008] The molecular weight of black fungus polysaccharide is 1K-10KDa.

[0009] Preferably, the molecular weight of the black fungus polysaccharide is ≤3KDa.

[0010] Preferably, the mass ratio of the bamboo leaf extract:green tea extract:black fungus polysaccharide is 0.5-1:1-2:5.

[0011] The white truffle fermented skin rejuvenation mask comprises, by weight, 1-5 parts of white truffle fermentation filtrate, 0.1-2 parts of bamboo leaf extract, 0.1-1 parts of green tea extract, 1-2 parts of black fungus polysaccharide, 1-1.5 parts of tremella extract, 1-3 parts of Bacillus fermentation product, 0.5-1.5 parts of wheat germ extract, 1-2 parts of Dendrobium officinale stem extract, 1-5 parts of glycerin, and 70-90 parts of water.

[0012] The preparation of the white truffle fermentation filtrate comprises the following steps:

[0013] S1. Powder the dried white truffles, and sieve through 200-400 mesh to obtain white truffle powder for later use;

[0014] S2, mixing white truffle powder and water in a certain weight ratio, adding papain for enzymolysis for 12-24 hours, inactivating the enzyme, and obtaining a white truffle enzymolysis solution;

[0015] S3, using the white truffle hydrolysate as a raw material, adding yeast and fermenting for 48-72 hours to obtain a white truffle fermentation product;

[0016] S4. Filtering the white truffle fermentation product and sterilizing it to obtain white truffle fermentation filtrate.

[0017] The bamboo leaf extract is an alcohol extract of bamboo leaves.

[0018] The green tea extract is a water extract of green tea.

[0019] In addition, the present invention discloses a method for preparing a preservative-free white truffle fermented skin rejuvenation mask, comprising the following steps:

[0020] 1) 1-5 parts of white truffle fermentation filtrate, 0.1-2 parts of bamboo leaf extract, 0.1-1 parts of green tea extract, 1-2 parts of black fungus polysaccharide, 1-1.5 parts of tremella extract, 1-3 parts of Bacillus fermentation product, 0.5-1.5 parts of wheat germ extract, 1-2 parts of Dendrobium officinale stem extract, 1-5 parts of glycerin and 70-90 parts of water are heated and stirred to obtain a facial mask liquid.

[0021] 2) The mask cloth and inner packaging materials are irradiated and sterilized, and the filling pipe equipped with a sterilizing-grade liquid filter is steam sterilized. After sterilization, the mask liquid is filled, sealed, inspected, and packaged in sequence.

[0022] The heating temperature is 60-85°C

[0023] The steam sterilization conditions are 110-121° C., 40-60 min.

[0024] Beneficial Effects

[0025] 1. The present invention uses yeast to ferment enzymatically hydrolyzed white truffles, thereby improving the extraction rate of protein in the white truffles. The obtained white truffle fermentation filtrate has the effect of promoting collagen synthesis and can be used for anti-aging functional cosmetics.

[0026] 2. The present invention utilizes the synergistic antibacterial effects of bamboo leaf extract, green tea extract, and black fungus polysaccharide in the white truffle fermentation filtrate mask to prepare a white truffle fermentation skin-rejuvenating mask that is natural, green, safe, and non-irritating. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Cytotoxicity test cell viability DETAILED DESCRIPTION

[0028] The technical solution of the present invention is further described below in conjunction with specific examples. The black fungus polysaccharide in the following examples was purchased from Xi'an Meihe Biotechnology Co., Ltd., and other raw materials used, unless otherwise specified, can be obtained from conventional commercial channels; the processes used, unless otherwise specified, all adopt conventional processes in the art.

[0029] Example 1

[0030] A method for preparing a white truffle fermentation filtrate comprises the following steps:

[0031] S1. Powder the dried white truffles and sieve through 200 mesh to obtain white truffle powder for later use;

[0032] S2, mixing white truffle powder and water in a weight ratio of 1:20, adding 1.5% papain (100,000 u / g) according to the weight of the white truffle powder, performing enzymolysis for 24 hours, inactivating the enzyme, and obtaining a white truffle enzymolysis solution;

[0033] S3, using white truffle hydrolysate as raw material, adding 1wt% yeast (the concentration of yeast in the white truffle hydrolysate is 3.75×10 8 CFU / mL) was subjected to anaerobic fermentation at 28°C for 72h to obtain white truffle fermentation products;

[0034] S4. Filtering the white truffle fermentation product and sterilizing it to obtain white truffle fermentation filtrate.

[0035] Example 2

[0036] A method for preparing a white truffle fermentation filtrate comprises the following steps:

[0037] S1. Powder the dried white truffles and sieve through 400 mesh to obtain white truffle powder for later use;

[0038] S2, mixing white truffle powder and water in a weight ratio of 1:20, adding 1.5% papain (100,000 u / g) according to the weight of the white truffle powder, performing enzymolysis for 24 hours, inactivating the enzyme, and obtaining a white truffle enzymolysis solution;

[0039] S3, using white truffle hydrolysate as raw material, adding 2wt% yeast (the concentration of yeast in the white truffle hydrolysate is 5.27×10 8 CFU / mL) was subjected to anaerobic fermentation at 28°C for 48h to obtain a white truffle fermentation product;

[0040] S4. Filtering the white truffle fermentation product and sterilizing it to obtain white truffle fermentation filtrate.

[0041] Example 3

[0042] The preparation of the plant extract comprises the following steps:

[0043] Preparation of green tea extract: according to the mass ratio of tea powder to water of 1:10, extraction at 75°C for 1 hour, filtration, to obtain the green tea extract.

[0044] Preparation of bamboo leaf water extract: according to the mass ratio of bamboo leaf powder to water of 1:10, extraction at 75°C for 1h, filtration, to obtain the bamboo leaf water extract.

[0045] Preparation of bamboo leaf alcohol extract: according to the mass ratio of bamboo leaf powder: water: glycerol of 1:4:6, extraction at 75°C for 1 hour, filtration, to obtain the bamboo leaf alcohol extract.

[0046] The preparation of black fungus polysaccharides with different molecular weights comprises the following steps:

[0047] The method comprises the following steps: preparing a black fungus polysaccharide solution according to a mass ratio of black fungus polysaccharide to water of 1:100, filtering with an ultrafiltration membrane with a molecular weight cutoff of 10 KDa to obtain a filtrate; filtering with an ultrafiltration membrane with a molecular weight cutoff of 5 KDa to obtain a retentate and a filtrate; freeze-drying the retentate to obtain a black fungus polysaccharide with a molecular weight of 5 KDa-10 KDa; filtering the filtrate with an ultrafiltration membrane with a molecular weight cutoff of 3 KDa to obtain a black fungus polysaccharide with a molecular weight of 3 KDa-5 KDa by freeze-drying the retentate, and freeze-drying the filtrate to obtain a black fungus polysaccharide with a molecular weight of less than 3 KDa.

[0048] Example 4

[0049] A preparation method of a preservative-free white truffle fermented skin rejuvenation mask comprises the following steps:

[0050] 1) Add 5 parts of white truffle fermentation filtrate, 0.1 parts of bamboo leaf extract, 0.2 parts of green tea extract, 0.5 parts of black fungus polysaccharide (molecular weight ≤ 3KDa), 1 part of tremella extract, 1 part of Bacillus fermentation product, 0.5 parts of wheat germ extract, 1 part of Dendrobium officinale stem extract, and 1 part of glycerin, add water to 100 parts, heat at 65°C and stir evenly to obtain a facial mask solution, and then sterilize it.

[0051] 2) The mask cloth and inner packaging materials are sterilized by irradiation, and the filling pipe equipped with a sterilizing-grade liquid filter is sterilized by steam at 121°C for 40 minutes. After sterilization, the mask liquid is filled, sealed, inspected, and packaged in sequence.

[0052] Example 5

[0053] A preparation method of a preservative-free white truffle fermented skin rejuvenation mask comprises the following steps:

[0054] 1) Add 1 part of white truffle fermentation filtrate, 1 part of bamboo leaf extract, 2 parts of green tea extract, 5 parts of black fungus polysaccharide (molecular weight ≤ 3KDa), 1.5 parts of tremella extract, 3 parts of Bacillus fermentation product, 1.5 parts of wheat germ extract, 2 parts of Dendrobium officinale stem extract, 5 parts of glycerin, add water to 100 parts, heat at 85°C and stir evenly to obtain a facial mask liquid, and then sterilize it.

[0055] 2) The mask cloth and inner packaging materials are irradiated and sterilized, and the filling pipe equipped with a sterilizing-grade liquid filter is steam sterilized at 110°C for 60 minutes. After sterilization, the mask liquid is filled, sealed, inspected, and packaged in sequence.

[0056] Comparative Example 1

[0057] A preparation method of a white truffle extract comprises the following steps:

[0058] S1. Powder the dried white truffles and sieve through 200 mesh to obtain white truffle powder for later use;

[0059] S2, mixing white truffle powder and water in a weight ratio of 1:20, adding 1.5% papain (100,000 u / g) according to the weight of the white truffle powder for enzymolysis for 24 hours to obtain a white truffle enzymolysis solution;

[0060] S3. Inactivate the enzyme in the white truffle enzymatic hydrolysate, and filter to obtain a white truffle extract.

[0061] Comparative Example 2

[0062] A method for preparing a white truffle fermentation filtrate comprises the following steps:

[0063] S1. Powder the dried white truffles and sieve through 200 mesh to obtain white truffle powder for later use;

[0064] S2, mixing white truffle powder and water in a weight ratio of 1:20 to obtain a white truffle mixture;

[0065] S3, using the white truffle mixture as raw material, adding 1wt% yeast (the concentration of yeast in the white truffle enzymatic hydrolysate is 3.75×10 8 CFU / mL) was subjected to anaerobic fermentation at 28°C for 72h to obtain white truffle fermentation products;

[0066] S4. Filtering the white truffle fermentation product and sterilizing it to obtain white truffle fermentation filtrate.

[0067] Comparative Example 3

[0068] A preparation method of a preservative-free white truffle fermented skin rejuvenation mask comprises the following steps:

[0069] 1) Add 1 part of white truffle fermentation filtrate, 1 part of bamboo leaf extract, 1.5 parts of Tremella fuciformis extract, 3 parts of Bacillus fermentation product, 1.5 parts of wheat germ extract, 2 parts of Dendrobium officinale stem extract, and 5 parts of glycerin, and add water to 100 parts, heat at 85° C. and stir evenly to obtain a facial mask liquid, and then sterilize it.

[0070] 2) The mask cloth and inner packaging materials are sterilized by irradiation, and the filling pipe equipped with a sterilizing-grade liquid filter is sterilized by steam at 121°C for 40 minutes. After sterilization, the mask liquid is filled, sealed, inspected, and packaged in sequence.

[0071] Comparative Example 4

[0072] A preparation method of a preservative-free white truffle fermented skin rejuvenation mask comprises the following steps:

[0073] 1) Add 1 part of white truffle fermentation filtrate, 1 part of bamboo leaf extract, 5 parts of black fungus polysaccharide (molecular weight ≤ 3KDa), 1.5 parts of tremella extract, 3 parts of Bacillus fermentation product, 1.5 parts of wheat germ extract, 2 parts of Dendrobium officinale stem extract, 5 parts of glycerin, and water to 100 parts, heat at 85°C and stir evenly to obtain a facial mask liquid, and then sterilize it.

[0074] 2) The mask cloth and inner packaging materials are sterilized by irradiation, and the filling pipe equipped with a sterilizing-grade liquid filter is sterilized by steam at 121°C for 40 minutes. After sterilization, the mask liquid is filled, sealed, inspected, and packaged in sequence.

[0075] Comparative Example 5

[0076] A preparation method of a preservative-free white truffle fermented skin rejuvenation mask comprises the following steps:

[0077] 1) Add 1 part of white truffle fermentation filtrate, 1 part of bamboo leaf extract, 2 parts of green tea extract, 1.5 parts of Tremella fuciformis extract, 3 parts of Bacillus fermentation product, 1.5 parts of wheat germ extract, 2 parts of Dendrobium officinale stem extract, and 5 parts of glycerin, add water to 100 parts, heat at 85° C. and stir evenly to obtain a facial mask liquid, and then sterilize it.

[0078] 2) The mask cloth and inner packaging materials are sterilized by irradiation, and the filling pipe equipped with a sterilizing-grade liquid filter is sterilized by steam at 121°C for 40 minutes. After sterilization, the mask liquid is filled, sealed, inspected, and packaged in sequence.

[0079] 1. Determination of protein content in white truffle fermentation filtrate or white truffle extract

[0080] The BCA kit method was used according to the operating steps in the instruction manual to determine the protein content in the white truffle fermentation filtrate and the white truffle extract in Examples 1 and 2 and Comparative Examples 1 and 2. The test results are shown in Table 1.

[0081] Table 1 Protein content in white truffle fermentation filtrate and white truffle extract

[0082] project Water-soluble protein content (mg / ml) Example 1 22.87 Example 2 25.45 Comparative Example 1 8.98 Comparative Example 2 12.32

[0083] 2. Cellular efficacy test of white truffle fermentation filtrate and white truffle extract

[0084] Collagen reconstruction is a key factor in maintaining skin vitality. Using human dermal fibroblasts to measure the content of type I collagen after administration can indirectly evaluate the skin rejuvenation effect of the white truffle fermentation filtrate and white truffle extract in Examples 1 and 2 and Comparative Examples 1 and 2.

[0085] ①MTT assay to test the cytotoxicity of active ingredients to human dermal fibroblasts

[0086] The cell suspension (100 μL / well, 10^ 4 Cells) were added to a 96-well plate and the plate was incubated in a humidified incubator for 24 h (37°C, 5% CO2). The culture medium was discarded, and the plate was incubated in a humidified incubator for 24 h (37°C, 5% CO2) after administration (5% white truffle fermentation filtrate or white truffle extract). 10 μL of CCK-8 solution was added to each well of the plate, and the plate was incubated in an incubator for 1-4 h. The absorbance at 450 nm was measured using an ELISA reader to calculate the cell survival rate. The results are shown in FIG. Figure 1 .

[0087] ② Determination of type I collagen content

[0088] The cell suspension (100 μL / well, 2*10^ 4 Cells) were added to a 96-well plate, and the plate was pre-cultured in a humid incubator for 24 h (37°C, 5% CO2). The culture medium was discarded, and the drug (1% white truffle fermentation filtrate or white truffle extract) was administered, and the plate was cultured in a humid incubator for 24 h (37°C, 5% CO2). After the incubation, the supernatant of the cells was collected, and the content of type I collagen was determined using a human type I collagen enzyme-linked immunosorbent assay kit. The results are shown in Table 2.

[0089] Table 2 Content of type I collagen

[0090] project Type I collagen content (ng / mg) blank 8.12 Example 1 40.66 Example 2 35.91 Comparative Example 1 20.12 Comparative Example 2 25.34

[0091] 3. Antibacterial performance test of plant raw materials and their compositions

[0092] Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Aspergillus niger, and Candida albicans were selected as test bacteria to test the minimum inhibitory concentration (MIC) of plant raw materials and their compositions, wherein composition 1 is bamboo leaf alcohol extract: green tea extract: black fungus polysaccharide (molecular weight ≤ 3KDa), composition 2 is bamboo leaf alcohol extract: green tea extract: black fungus polysaccharide (molecular weight 3KDa-5KDa), and composition 3 is bamboo leaf alcohol extract: green tea extract: black fungus polysaccharide (molecular weight 5K-10KDa). Among them, the mass ratio of bamboo leaf alcohol extract: green tea extract: black fungus polysaccharide in compositions 1-3 is 1:2:5, and the test results are shown in Table 3.

[0093] Table 3 Minimum inhibitory concentration of antibacterial substances

[0094]

[0095] IV. White Truffle Fermented Skin Rejuvenation Mask Antiseptic Challenge Test

[0096] With reference to the United States Pharmacopoeia-antiseptic and antibacterial ability test, the white truffle fermented skin rejuvenation mask in Examples 4 and 5 and Comparative Examples 3-5 was subjected to an antiseptic challenge test. The test results are shown in Tables 4 and 5.

[0097] Table 4 Bacterial antiseptic challenge results

[0098]

[0099] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A preservative-free white truffle fermented skin rejuvenation mask, characterized in that: It includes white truffle fermentation filtrate, bamboo leaf extract, green tea extract and black fungus polysaccharide; the molecular weight of the black fungus polysaccharide is 1K-10KDa.

2. The white truffle fermented skin rejuvenation mask according to claim 1, characterized in that: The invention comprises white truffle fermentation filtrate, bamboo leaf extract, green tea extract and black fungus polysaccharide, wherein the molecular weight of the black fungus polysaccharide is less than or equal to 3KDa.

3. The white truffle fermented skin rejuvenation mask according to claim 2, characterized in that: The mass ratio of the bamboo leaf extract, the green tea extract and the black fungus polysaccharide is 0.5-1:1-2:

5.

4. The white truffle fermented skin rejuvenation mask according to claim 3, characterized in that: The invention comprises 1-5 parts of white truffle fermentation filtrate, 0.1-2 parts of bamboo leaf extract, 0.1-1 parts of green tea extract, 1-2 parts of black fungus polysaccharide, 1-1.5 parts of tremella extract, 1-3 parts of bacillus fermentation product, 0.5-1.5 parts of wheat germ extract, 1-2 parts of Dendrobium officinale stem extract, 1-5 parts of glycerin and 70-90 parts of water.

5. The white truffle fermented skin rejuvenation mask according to claim 4, characterized in that: The preparation of the white truffle fermentation filtrate comprises the following steps: S1. Powder the dried white truffles, and sieve through 200-400 mesh to obtain white truffle powder for later use; S2, mixing white truffle powder and water in a certain weight ratio, adding papain for enzymolysis for 12-24 hours, and obtaining a white truffle enzymolysis solution; S3, using the white truffle hydrolysate as a raw material, adding yeast and fermenting for 48-72 hours to obtain a white truffle fermentation product; S4. Filtering the white truffle fermentation product and sterilizing it to obtain white truffle fermentation filtrate.

6. The white truffle fermented skin rejuvenation mask according to claim 5, characterized in that: The bamboo leaf extract is an alcohol extract of bamboo leaves.

7. The white truffle fermented skin rejuvenation mask according to claim 6, characterized in that: The green tea extract is a water extract of green tea.

8. A method for preparing a preservative-free white truffle fermented skin rejuvenation mask, characterized in that: The following steps are involved: 1) heating and stirring 1-5 parts of white truffle fermentation filtrate, 0.1-2 parts of bamboo leaf extract, 0.1-1 parts of green tea extract, 1-2 parts of black fungus polysaccharide, 1-1.5 parts of tremella extract, 1-3 parts of Bacillus fermentation product, 0.5-1.5 parts of wheat germ extract, 1-2 parts of Dendrobium officinale stem extract, 1-5 parts of glycerin and 70-90 parts of water to obtain a facial mask liquid; 2) The mask cloth and inner packaging materials are irradiated and sterilized, and the filling pipe equipped with a sterilizing-grade liquid filter is steam sterilized. After sterilization, the mask liquid is filled, sealed, inspected, and packaged in sequence.

9. The method for preparing the white truffle fermented skin rejuvenation mask according to claim 8, characterized in that: The heating temperature is 60-85°C.

10. The method for preparing the white truffle fermented skin rejuvenation mask according to claim 9, characterized in that: The steam sterilization conditions are 110-121° C., 40-60 min.

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