A preservative-free white truffle fermented skin-nourishing mask and its preparation method

Abstract

The present invention relates to a preservative-free white truffle fermented skin-nourishing mask and a preparation method thereof. The preservative-free white truffle fermented skin-nourishing mask comprises 1-5 parts of white truffle fermentation filtrate, 0.1-2 parts of bamboo leaf extract, 0.1-1 part of green tea extract, 1-2 parts of auricularia 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 glycerol, and 70-90 parts of water. The present invention uses papain to enzymatically hydrolyze white truffle, and then ferments it with yeast to obtain a white truffle fermentation filtrate with a high water-soluble protein content, and adds bamboo leaf extract, green tea extract, and auricularia polysaccharide to the fermentation filtrate to prepare a green preservative-free white truffle fermented skin-nourishing mask.

Description

Technical Field

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

[0002] A mask is a beauty care product, which has become an essential item for women's skin care after years of development. The main function of the mask is to penetrate nutrients into the skin through the film material, so as to achieve the effect of skin care. However, the moisture and nutrients in the mask are easily contaminated by microorganisms such as bacteria and molds, resulting in deterioration. Without adding preservatives, the mask is easily infected by microorganisms, producing peculiar smells, spoilage, and even seriously affecting the skin health of users. And long-term use of preservatives has potential harm to the skin, including allergies, irritation, dryness, and damage to the skin barrier, etc.

[0003] White truffle is a kind of tuber mushroom, containing crude protein, fat, amino acids, polyphenols, polysaccharides, various 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 ingredient, but also has high medicinal value, and is called the "king of immunity" together with Ganoderma lucidum. White truffle also has a variety of bioactive components, which can not only inhibit the production of melanin, thus achieving the effect of whitening and brightening the skin; but also has antioxidant and anti-aging activities, and can activate the vitality of cells to regenerate.

[0004] The invention patent with the publication number of CN114748381A discloses a method for preparing white truffle fermentation broth, which uses white truffle extract as the raw material and ferments it with Bacillus. By using the water extraction method, the obtained white truffle extract improves the bioavailability of white truffle. In order to improve the bioavailability of white truffle and at the same time improve the safety of using white truffle, it is urgent to develop a preservative-free white truffle fermented skin-nourishing mask and a preparation method thereof. Summary of the Invention

[0005] In order to solve the above technical problems, the present invention provides a preservative-free white truffle fermented skin-nourishing mask and a preparation method thereof. Papain is used to enzymatically hydrolyze white truffle, and then yeast is used for fermentation to obtain a white truffle fermentation filtrate with a high content of water-soluble protein. And bamboo leaf extract, green tea extract and Auricularia auricula polysaccharide are added to the fermentation filtrate to prepare a preservative-free white truffle fermented skin-nourishing mask.

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

[0007] A preservative-free white truffle fermented skin-nourishing mask, comprising: white truffle fermentation filtrate, bamboo leaf extract, green tea extract, Auricularia auricula polysaccharide, calculated by mass parts.

[0008] As described above, the molecular weight of the Auricularia auricula polysaccharide is 1K - 10KDa.

[0009] Preferably, the molecular weight of the Auricularia auricula polysaccharide ≤ 3KDa.

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

[0011] As described above, the white truffle fermented skin - smoothing mask, by weight, comprises 1 - 5 parts of white truffle fermentation filtrate, 0.1 - 2 parts of bamboo leaf extract, 0.1 - 1 part of green tea extract, 1 - 2 parts of Auricularia auricula polysaccharide, 1 - 1.5 parts of Tremella fuciformis 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 glycerol, 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 truffle, and sieve it through a 200 - 400 mesh sieve to obtain white truffle powder for standby;

[0014] S2. Mix the white truffle powder with water according to a certain weight ratio, add papain for enzymatic hydrolysis for 12 - 24h, inactivate the enzyme, and obtain the white truffle enzymatic hydrolysate;

[0015] S3. Using the white truffle enzymatic hydrolysate as the raw material, add yeast for fermentation for 48 - 72h to obtain the white truffle fermentation product;

[0016] S4. Filter the white truffle fermentation product, and after sterilization, obtain the white truffle fermentation filtrate.

[0017] As described above, the bamboo leaf extract is the alcohol extract of bamboo leaves.

[0018] As described above, the green tea extract is the water extract of green tea.

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

[0020] 1) Heat and stir evenly 1 - 5 parts of white truffle fermentation filtrate, 0.1 - 2 parts of bamboo leaf extract, 0.1 - 1 part of green tea extract, 1 - 2 parts of Auricularia auricula polysaccharide, 1 - 1.5 parts of Tremella fuciformis 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 glycerol, and 70 - 90 parts of water to obtain the mask liquid.

[0021] 2) Irradiate and sterilize the facial mask cloth and the inner packaging material, and perform steam sterilization on the filling pipeline equipped with a sterilizing-grade liquid filter. After sterilization, fill, seal, inspect, and package the facial mask liquid in sequence.

[0022] As described above, the heating temperature is 60 - 85 °C

[0023] As described above, the conditions for steam sterilization are 110 - 121 °C and 40 - 60 min.

[0024] Beneficial effects

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

[0026] 2. The present invention utilizes the synergistic antibacterial effect of bamboo leaf extract, green tea extract, and auricularia auricula polysaccharide in the white truffle fermentation filtrate facial mask, and the prepared white truffle fermented skin-nourishing facial mask is natural, green, safe, and non-irritating. Description of the drawings

[0027] Figure 1 is the cell survival rate in the cytotoxicity test Specific implementation manners

[0028] The technical solutions of the present invention are further described below in conjunction with specific embodiments. The auricularia auricula polysaccharide in the following embodiments is purchased from Xi'an Meihe Biotechnology Co., Ltd. Other raw materials used, unless otherwise specified, can be obtained from conventional commercial channels; the processes adopted, unless otherwise specified, are conventional processes in the art.

[0029] Example 1

[0030] The preparation of a white truffle fermentation filtrate includes the following steps:

[0031] S1. Powder the dried white truffle and sieve it through a 200-mesh sieve to obtain white truffle powder for standby;

[0032] S2. Mix the white truffle powder with water at a weight ratio of 1:20, and add 1.5% papain (100,000 u / g) according to the weight of the white truffle powder for enzymatic hydrolysis for 24 h, and inactivate the enzyme to obtain a white truffle enzymatic hydrolysate;

[0033] S3. Using the white truffle enzymatic hydrolysate as the raw material, add 1 wt% yeast (the concentration of yeast in the white truffle enzymatic hydrolysate is 3.75×10 8 CFU / mL) for anaerobic fermentation at 28 °C for 72 h to obtain a white truffle fermentation product;

[0034] S4. Filter the white truffle fermentation product, and after sterilization, obtain the white truffle fermentation filtrate.

[0035] Example 2

[0036] Preparation of a white truffle fermentation filtrate, comprising the following steps:

[0037] S1. Powder the dried white truffle, and sieve it through a 400-mesh sieve to obtain white truffle powder for standby;

[0038] S2. Mix the white truffle powder and water according to a weight ratio of 1:20, and add 1.5% papain (100,000 u / g) based on the weight of the white truffle powder for enzymatic hydrolysis for 24 h, inactivate the enzyme, and obtain the white truffle enzymatic hydrolysate;

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

[0040] S4. Filter the white truffle fermentation product, and after sterilization, obtain the white truffle fermentation filtrate.

[0041] Example 3

[0042] Preparation of a plant extract, comprising the following steps:

[0043] Preparation of green tea extract: According to the mass ratio of tea powder to water of 1:10, extract at 75°C for 1 h, and filter 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, extract at 75°C for 1 h, and filter to obtain the bamboo leaf water extract.

[0045] Preparation of bamboo leaf alcohol extract: According to the mass ratio of bamboo leaf powder to water to glycerol of 1:4:6, extract at 75°C for 1 h, and filter to obtain the bamboo leaf alcohol extract.

[0046] Preparation of black fungus polysaccharides with different molecular weights, comprising the following steps:

[0047] Prepare an auricularia polysaccharide solution according to the mass ratio of auricularia polysaccharide to water of 1:100, filter it using an ultrafiltration membrane with a molecular weight cut-off of 10 KDa to obtain a filtrate; then filter the filtrate using an ultrafiltration membrane with a molecular weight cut-off of 5 KDa to obtain a retentate and a filtrate; lyophilize the retentate to obtain auricularia polysaccharide with a molecular weight of 5 KDa - 10 KDa; filter the filtrate again using an ultrafiltration membrane with a molecular weight cut-off of 3 KDa, lyophilize the retentate to obtain auricularia polysaccharide with a molecular weight of 3 KDa - 5 KDa, and lyophilize the filtrate to obtain auricularia polysaccharide with a molecular weight of less than 3 KDa.

[0048] Example 4

[0049] Preparation of a preservative-free white truffle fermented skin-nourishing mask, comprising the following steps:

[0050] 1) Mix 5 parts of white truffle fermentation filtrate, 0.1 part of bamboo leaf extract, 0.2 part of green tea extract, 0.5 part of auricularia polysaccharide (molecular weight ≤ 3 KDa), 1 part of tremella extract, 1 part of bacillus fermentation product, 0.5 part of wheat germ extract, 1 part of dendrobium officinale stem extract, 1 part of glycerol, and add water to make 100 parts. Heat and stir evenly at 65 °C to obtain a mask liquid, and then perform sterilization treatment.

[0051] 2) Perform irradiation sterilization on the mask cloth and inner packaging materials, perform steam sterilization at 121 °C for 40 min on the filling pipeline equipped with a sterile liquid filter. After sterilization, fill, seal, inspect, and package the mask liquid in sequence.

[0052] Example 5

[0053] Preparation of a preservative-free white truffle fermented skin-nourishing mask, comprising the following steps:

[0054] 1) Mix 1 part of white truffle fermentation filtrate, 1 part of bamboo leaf extract, 2 parts of green tea extract, 5 parts of auricularia polysaccharide (molecular weight ≤ 3 KDa), 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 glycerol, and add water to make 100 parts. Heat and stir evenly at 85 °C to obtain a mask liquid, and then perform sterilization treatment.

[0055] 2) Perform irradiation sterilization on the mask cloth and inner packaging materials, perform steam sterilization at 110 °C for 60 min on the filling pipeline equipped with a sterile liquid filter. After sterilization, fill, seal, inspect, and package the mask liquid in sequence.

[0056] Comparative Example 1

[0057] Preparation of a white truffle extract, comprising the following steps:

[0058] S1. Powder the dried white truffles and sieve through a 200-mesh sieve to obtain white truffle powder for standby.

[0059] S2. Mix the white truffle powder with water at a weight ratio of 1:20. Based on the weight of the white truffle powder, add 1.5% papain (100,000 u / g) and carry out enzymatic hydrolysis for 24 h to obtain white truffle enzymatic hydrolysate.

[0060] S3. Inactivate the enzyme in the white truffle enzymatic hydrolysate, and after filtration, obtain white truffle extract.

[0061] Comparative Example 2

[0062] The preparation of a white truffle fermentation filtrate includes the following steps:

[0063] S1. Powder the dried white truffles and sieve through a 200-mesh sieve to obtain white truffle powder for standby.

[0064] S2. Mix the white truffle powder with water at a weight ratio of 1:20 to obtain white truffle mixture.

[0065] S3. Using the white truffle mixture as raw material, based on the weight of the white truffle powder, add 1 wt% yeast (the concentration of yeast in the white truffle enzymatic hydrolysate is 3.75×10 8 CFU / mL) and carry out anaerobic fermentation at 28 °C for 72 h to obtain white truffle fermentation product.

[0066] S4. Filter the white truffle fermentation product, and after sterilization, obtain white truffle fermentation filtrate.

[0067] Comparative Example 3

[0068] The preparation of a preservative-free white truffle fermentation skin-nourishing mask includes the following steps:

[0069] 1) Mix 1 part of white truffle fermentation filtrate, 1 part of bamboo leaf extract, 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 glycerol, and add water to 100 parts. Heat and stir evenly at 85 °C to obtain mask liquid, and then carry out sterilization treatment.

[0070] 2) Carry out irradiation sterilization on the mask cloth and inner packaging material, carry out steam sterilization at 121 °C for 40 min on the filling pipeline equipped with a sterilizing-grade liquid filter. After sterilization is completed, fill, seal, inspect, and package the mask liquid in sequence.

[0071] Comparative Example 4

[0072] The preparation of a preservative-free white truffle fermentation skin-nourishing mask includes the following steps:

[0073] 1) Mix 1 part of white truffle fermentation filtrate, 1 part of bamboo leaf extract, 5 parts of auricularia 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 add water to 100 parts. Heat and stir evenly at 85°C to obtain the facial mask liquid, and then perform sterilization treatment.

[0074] 2) Irradiate and sterilize the facial mask cloth and inner packaging materials, and perform steam sterilization at 121°C for 40 minutes on the filling pipeline equipped with a sterile liquid filter. After sterilization, fill, seal, inspect, and package the facial mask liquid in sequence.

[0075] Comparative Example 5

[0076] Preparation of a preservative-free white truffle fermented skin-nourishing facial mask, including the following steps:

[0077] 1) Mix 1 part of white truffle fermentation filtrate, 1 part of bamboo leaf extract, 2 parts of green tea extract, 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 add water to 100 parts. Heat and stir evenly at 85°C to obtain the facial mask liquid, and then perform sterilization treatment.

[0078] 2) Irradiate and sterilize the facial mask cloth and inner packaging materials, and perform steam sterilization at 121°C for 40 minutes on the filling pipeline equipped with a sterile liquid filter. After sterilization, fill, seal, inspect, and package the facial mask liquid in sequence.

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

[0080] Using the BCA kit method and following the operation steps in the instruction manual, determine the protein content in the white truffle fermentation filtrate and white truffle extract in Examples 1, 2 and Comparative Examples 1, 2. The test results are shown in Table 1.

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

[0082] Item Content of water-soluble protein (mg / ml) Example 1 22.87 Example 2 25.45 Comparative Example 1 8.98 Comparative Example 2 12.32

[0083] II. Cell 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, measuring the content of type I collagen after administration can indirectly evaluate the skin-nourishing efficacy of the white truffle fermentation filtrate and white truffle extract in Examples 1, 2 and Comparative Examples 1, 2.

[0085] ① Use the MTT method to test the cytotoxicity of the active ingredient on human dermal fibroblasts

[0086] Add cell suspension (100 μL / well, 10^ 4 cells per well) to a 96-well plate and incubate the plate in a humidified incubator for 24 h (37 °C, 5% CO2). Discard the culture medium, administer the drug (5% white truffle fermentation filtrate or white truffle extract), and then incubate the plate in a humidified incubator for 24 h (37 °C, 5% CO2). Add 10 μL of CCK-8 solution to each well of the plate, incubate the plate in the incubator for 1 - 4 h, measure the absorbance at 450 nm using a microplate reader, calculate the cell survival rate, and the results are as Figure 1 .

[0087] ② Determine the content of type I collagen

[0088] Add cell suspension (100 μL / well, 2*10^ 4 cells per well) to a 96-well plate and pre-incubate the plate in a humidified incubator for 24 h (37 °C, 5% CO2). Discard the culture medium, administer the drug (1% white truffle fermentation filtrate or white truffle extract), incubate the plate in a humidified incubator for 24 h (37 °C, 5% CO2). After incubation, collect the cell supernatant and use a human type I collagen ELISA kit to determine the content of type I collagen. The results are shown in Table 2.

[0089] Table 2 Content of type I collagen

[0090] Item Content of type I collagen (ng / mg) Blank 8.12 Example 1 40.66 Example 2 35.91 Comparative Example 1 20.12 Comparative Example 2 25.34

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

[0092] Select Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Aspergillus niger, and Candida albicans as test bacteria to test the minimum inhibitory concentration (MIC) of plant raw materials and their compositions. Among them, Composition 1 is bamboo leaf alcohol extract: green tea extract: Auricularia auricula polysaccharide (molecular weight ≤ 3 kDa), Composition 2 is bamboo leaf alcohol extract: green tea extract: Auricularia auricula polysaccharide (molecular weight 3 kDa - 5 kDa), and Composition 3 is bamboo leaf alcohol extract: green tea extract: Auricularia auricula polysaccharide (molecular weight 5K - 10KDa). Among them, the mass ratio of bamboo leaf alcohol extract: green tea extract: Auricularia auricula 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. Antiseptic challenge test of white truffle fermented skin rejuvenating mask

[0096] Refer to the Antimicrobial and Antibacterial Potency Test of the United States Pharmacopeia, and conduct an antimicrobial challenge test on the white truffle fermented skin rejuvenating mask in Examples 4 and 5 and Comparative Examples 3-5. The test results are shown in Tables 4 and 5.

[0097] Table 4 Results of Bacterial Antimicrobial Challenge

[0098]

[0099] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. A preservative-free white truffle fermented skin rejuvenation mask, characterized in that: Includes white truffle fermentation filtrate, bamboo leaf extract, green tea extract, black fungus polysaccharide; Wherein, 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 with water, adding papain for enzymolysis for 12-24 hours, and obtaining 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 a white truffle fermentation filtrate; the bamboo leaf extract is an alcohol extract of bamboo leaves; The green tea extract is a water extract of green tea; The mass ratio of the bamboo leaf extract, the green tea extract and the black fungus polysaccharide is 0.5-1:1-2:5; 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 molecular weight of the black fungus polysaccharide is ≤3KDa.

3. The white truffle fermented skin rejuvenation mask according to claim 2, 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.

4. A method for preparing the white truffle fermented skin rejuvenation mask according to claim 3, characterized in that: The following steps are involved: 1) Heat and stir 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 solution; 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.

5. The method for preparing the white truffle fermented skin rejuvenation mask according to claim 4, characterized in that: The heating temperature is 60-85°C.

6. The method for preparing the white truffle fermented skin rejuvenation mask according to claim 5, characterized in that: The steam sterilization conditions are 110-121° C., 40-60 min.

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