Compact and anti-wrinkle composition of multi-type recombinant collagen and application of compact and anti-wrinkle composition

Abstract

The invention belongs to the technical field of biology and new medicine, and particularly relates to a tightening and anti-wrinkle composition of multi-type recombinant collagen and application of the tightening and anti-wrinkle composition. The firming and anti-wrinkle composition is prepared from the following raw materials in percentage by weight: 0.5 to 5 percent of inositol, 0.1 to 0.5 percent of carnosine, 0.02 to 0.15 percent of collagen, 0.003 to 0.15 percent of citrus peel extract, 0.4 to 0.7 percent of firming composition, 0.1 to 0.4 percent of anti-wrinkle composition, 0.01 to 0.05 percent of butanediol, 0.002 to 0.009 percent of p-hydroxyacetophenone and the balance of water. The prepared firming and anti-wrinkle composition has excellent firming, anti-wrinkle and whitening effects at the same time, and is mild and non-irritant.

Description

Technical Field

[0001] The invention belongs to the field of biology and new medicine technology, and specifically relates to a firming and anti-wrinkle composition of polytype recombinant collagen and application thereof. Background Art

[0002] As we age, the collagen and elastic fibers in our skin gradually disappear, causing our skin to lose its elasticity and firmness, making it more prone to wrinkles and sagging. There are many firming and anti-wrinkle cosmetics on the market, but there are not many truly effective and gentle products.

[0003] Collagen is extracted from animal skins. In addition to collagen, the skins also contain proteoglycans such as hyaluronic acid and chondroitin sulfate. These contain a large number of polar groups and are moisturizing factors. They also have the function of preventing tyrosine in the skin from converting into melanin. Therefore, collagen has moisturizing, whitening, anti-wrinkle, and freckle removal effects, and can be widely used in beauty products. Although the firming and anti-wrinkle products containing collagen on the market are very popular, there are also some problems: (1) Collagen absorption problem: Collagen is a large molecular protein. Ordinary collagen is difficult to be directly absorbed by the skin. Even if applied to the surface of the skin, most of it will remain in the stratum corneum and cannot penetrate into the dermis to exert its effect. (2) Stability: Collagen is easily denatured under conditions such as high temperature and light, which affects its biological activity. Therefore, how to maintain the stability of collagen in the product is a challenge. (3) The effect is not significant: Many products containing collagen can only provide short-term moisturizing and firming effects, and the anti-wrinkle effect after long-term use is not obvious. (4) Allergic reactions: Some people may have allergic reactions to collagen or other ingredients in the product, such as redness, swelling, stinging, etc. (5) Insufficient efficacy: As consumers' demands become increasingly stringent, the efficacy requirements for a skin care product are also getting higher and higher, which will directly affect the market competitiveness of the product.

[0004] The Chinese invention patent with the announcement number CN116270295B provides an anti-wrinkle composition containing whole human collagen and its application, wherein the components of the anti-wrinkle composition include whole human collagen, an amino acid composition, acetyl hexapeptide-8 and dipeptide diaminobutyryl benzylamide diacetate. The composition provided by the method improves the anti-wrinkle effect of the product through the synergistic combination of active ingredients such as whole human collagen, an amino acid composition, acetyl hexapeptide-8 and dipeptide diaminobutyryl benzylamide diacetate. However, the long-term use effect of the above anti-wrinkle composition is not good. Summary of the invention

[0005] The purpose of the present invention is to provide a firming and anti-wrinkle composition of multi-type recombinant collagen and its application.

[0006] In order to achieve the above object, the present invention provides the following technical solutions: A firming and anti-wrinkle composition of multi-type recombinant collagen, comprising the following raw materials in percentage by weight: 0.5-5% inositol, 0.1-0.5% carnosine, 0.02-0.15% collagen, 0.003-0.15% citrus (CITRUSRETICULATA) peel extract, 0.4-0.7% firming composition, 0.1-0.4% anti-wrinkle composition, 0.01-0.05% butylene glycol, 0.002-0.009% p-hydroxyacetophenone, and the balance is water; The preparation method of the firming composition comprises the following steps: (1) Sichuan red peony root ( Paonia Veitchii ) roots, wild sycamore ( Mallotus japonicus ) bark, Passiflora incarnata ( Passiflora Quadrangularis ) and the fruits are mixed, and crushed to less than 60 meshes after mixing to obtain a first mixed raw material; (2) placing the first mixed raw material at a low temperature of -28°C for 2-3 minutes, then rapidly heating it to 25-27°C within 2 seconds, repeating this process 2-3 times; then pressurizing it to 18-20 MPa, maintaining it for 5-6 minutes, then releasing the pressure to normal pressure, repeating this process 2-3 times, and obtaining a pretreated first mixed raw material; (3) Mixing the pretreated first mixed raw material and water in a weight ratio of 1: (10-15) to obtain a first fermentation raw material, adding composite bacteria to the first fermentation raw material, wherein the composite bacteria includes Lactobacillus acidophilus, Bifidobacterium longum and Lactobacillus helveticus, and the amount of Lactobacillus acidophilus is 10 5 -10 6 CFU / mL first fermentation raw material; the dosage of Bifidobacterium longum is 10 7 -10 8 CFU / mL first fermentation raw material; the amount of Lactobacillus helveticus is 10 6 -10 7 CFU / mL first fermentation raw material; ferment at 38-40°C for 8-10h, sterilize, and obtain a first fermentation product; (4) The first fermentation product is filtered using 0.45 μm and 0.22 μm filter membranes respectively, and spray-dried to obtain a firming composition.

[0007] Furthermore, the weight ratio of the root of Paeonia lactiflora, the bark of the wild sycamore tree, and the large-fruited passion fruit is 1:(0.3-0.5):(1.5-1.7).

[0008] The present invention can improve the firming effect of the firming and anti-wrinkle composition by adding a firming composition. Mainly, the paeoniflorin in the red peony root can activate fibroblasts, promote the synthesis of collagen and elastic fibers, and thus improve skin elasticity. The tannins and flavonoids in the bark of the wild sycamore tree have an antioxidant effect and can protect the skin from damage by free radicals; the natural moisturizing factor in the large-fruit passion fruit can increase the moisture content of the skin and make the skin softer and firmer; through fermentation, the firming composition contains a variety of active components that work synergistically and can promote collagen to better perform its physiological functions.

[0009] Furthermore, the preparation method of the anti-wrinkle composition comprises the following steps: (1) Desert verbena ( Abronia Villosa ) leaves, Polygonatum yunnanensis (P Kingianum olygonatum ) and sword bean ( Canavalia Gladiata ) seeds are mixed and crushed to less than 60 meshes to obtain a second mixed raw material; (2) Mixing the second mixed raw material and water in a weight ratio of 1: (8-10) to obtain a second fermentation raw material; adding alkaline protease in an amount of 3000-4000u / mL of the second fermentation raw material, performing enzymatic hydrolysis at 38-40°C for 30-40min, inactivating the enzyme, continuing to add papain in an amount of 2000-3000u / mL of the second fermentation raw material, continuing to perform enzymatic hydrolysis at 38-40°C for 80-90min, inactivating the enzyme, and finally adding neutral protease in an amount of 4000-5000u / mL of the second fermentation raw material, performing enzymatic hydrolysis at 38-40°C for 50-60min, inactivating the enzyme, and cooling to room temperature to obtain a pretreated second fermentation raw material; (3) Add animal Bifidobacterium to the pretreated second fermentation raw material, the amount of animal Bifidobacterium is 10 6 -10 7 CFU / mL second fermentation raw material; ferment at 38-40℃ for 5-7h, sterilize, and continue to add Lactobacillus casei, the amount of Lactobacillus casei is 10 8 -10 9 CFU / mL second fermentation raw material, ferment at 38-40℃ for 3-5h, sterilize, and finally add Bifidobacterium bifidum, the dosage of Bifidobacterium bifidum is 10 7 -10 8 CFU / mL second fermentation raw material, ferment at 38-40℃ for 2-4h, sterilize to obtain the second fermentation product, (4) The second fermentation product is filtered using 0.45 μm and 0.22 μm filter membranes respectively, and spray-dried to obtain an anti-wrinkle composition.

[0010] Furthermore, the weight ratio of the desert verbena leaves, polygonatum cyrtonema and sword bean seeds is 1: (1.3-1.5): (0.5-0.7).

[0011] The present invention can improve the anti-wrinkle effect of the firming anti-wrinkle composition by adding the anti-wrinkle composition. The macromolecular active substances are first decomposed into small molecules by enzymatic hydrolysis, and new active substances such as antioxidant substances may be produced in the fermentation process to further enhance the antioxidant capacity. At the same time, new polypeptides and amino acids may be produced in the fermentation process, and these substances have moisturizing and anti-wrinkle effects. At the same time, fermentation can improve the stability of certain active ingredients, making them less likely to be inactivated during storage and use.

[0012] Furthermore, the collagen is selected from one or more of recombinant type I humanized collagen, recombinant type III humanized collagen, and recombinant type XVII humanized collagen.

[0013] Furthermore, the collagen comprises a mixture of recombinant type I humanized collagen, recombinant type III humanized collagen and recombinant type XVII humanized collagen in a weight ratio of 1: (1.3-1.5): (0.6-0.8).

[0014] Furthermore, the weight ratio of collagen, firming composition and anti-wrinkle composition is (0.8-1):(5-6):(2-3).

[0015] When the collagen of the present invention is mixed with recombinant humanized collagen type I, recombinant humanized collagen type III and recombinant humanized collagen type XVII in a specific ratio, and compounded with a firming composition and an anti-wrinkle composition in a specific ratio, the whitening effect of the firming and anti-wrinkle composition can be improved. This is mainly because the components in the collagen, the firming composition and the anti-wrinkle composition work synergistically, which can promote the absorption of each other's active components and achieve better firming, anti-wrinkle and whitening effects.

[0016] Furthermore, the invention comprises the following raw materials in percentage by weight: 1% inositol, 0.3% carnosine, 0.09% collagen, 0.11% citrus peel extract, 0.54% firming composition, 0.23% anti-wrinkle composition, 0.04% butylene glycol, 0.006% p-hydroxyacetophenone, and the balance is water.

[0017] Furthermore, the preparation method of the firming and anti-wrinkle composition of multi-type recombinant collagen comprises the following steps: Step 1: Clean and disinfect equipment; Step 2: Add water to the emulsifier, start stirring, heat to 80-85℃ and keep warm for 30-40 minutes, then start cooling down; Step 3: Stop cooling when the temperature drops to 60°C; add a mixture of butanediol and p-hydroxyacetophenone preheated to 60-65°C, stir until uniform, and then start cooling; Step 4: Cool down, add inositol, and stir until uniform; Step 5: Cool down, add carnosine, and stir until uniform; Step 6: Cool down, add collagen, and stir until uniform; Step 7: Cool down, add remaining ingredients, and stir until uniform; Step 8: After cooling and passing the microbiological test, the product is bottled and then tested for microorganisms again to be the finished product.

[0018] The present invention provides application of the firming and anti-wrinkle composition of polytype recombinant collagen in the preparation of cosmetics.

[0019] Compared with the prior art, the advantages and beneficial effects of the present invention are: 1. The present invention combines recombinant type I collagen and recombinant type III collagen in a scientific and reasonable ratio with recombinant type XVII collagen to achieve stronger firming and anti-wrinkle effects. It combines anti-oxidation with firming and anti-wrinkle effects to make the skin more translucent.

[0020] 2. The present invention can improve the firming effect of the firming and anti-wrinkle composition by adding a firming composition.

[0021] 3. The present invention can improve the anti-wrinkle effect of the firming anti-wrinkle composition by adding the anti-wrinkle composition.

[0022] 4. The collagen of the present invention is a mixture of recombinant humanized collagen type I, recombinant humanized collagen type III and recombinant humanized collagen type XVII in a specific ratio, and is compounded with a firming composition and an anti-wrinkle composition in a specific ratio, which can improve the whitening effect of the firming and anti-wrinkle composition. At the same time, it can also improve the firming and anti-wrinkle effect. DETAILED DESCRIPTION

[0023] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0024] The raw materials used in the following examples of the present invention are all commercially available products: Lactobacillus acidophilus, number: CCTCC AB 2010208. was purchased from China Center for Type Culture Collection.

[0025] Bifidobacterium animalis, number: SHBCC D24407AS 1.1852, was purchased from China Center for Type Culture Collection.

[0026] Lactobacillus casei, number: SHBCC D24737, was purchased from Shanghai Microbiological Collection Center.

[0027] Bifidobacterium bifidum, number: SHBCC D24408 ATCC 35914, was purchased from Shanghai Microbiological Collection Center.

[0028] Lactobacillus helveticus was purchased from China Center for Type Culture Collection, with the number: CCTCCAB2010205.

[0029] Bifidobacterium longum was purchased from Shanghai Collection of Microorganisms Center, with the number: SHBCC D24310.

[0030] The enzymes used in the present invention were all purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0031] Carnosine: L-Carnosine, Hebei Qiansheng Biotechnology Co., Ltd.

[0032] Collagen: Fish skin collagen, fully water-soluble, extracted from tilapia skin, Shandong Lezhiwei Biotechnology Co., Ltd.

[0033] Citrus peel extract: oil-soluble, Wuhan Huaxiang Kejie Biotechnology Co., Ltd.

[0034] Example 1 The present embodiment provides a firming and anti-wrinkle composition of polytype recombinant collagen, comprising the following raw materials in weight percentage: 1% inositol, 0.3% carnosine, 0.09% collagen, 0.11% citrus peel extract, 0.54% firming composition, 0.23% anti-wrinkle composition, 0.04% 1,3-butanediol, 0.006% p-hydroxyacetophenone, and the balance is water.

[0035] The collagen comprises a mixture of recombinant type I humanized collagen, recombinant type III humanized collagen and recombinant type XVII humanized collagen in a weight ratio of 1:1.4:0.7.

[0036] The preparation method of the firming composition comprises the following steps: (1) mixing the root of Paeonia lactiflora, the bark of Pseudostellaria baicalensis, and the large-fruited passion fruit in a weight ratio of 1:0.4:1.6, and crushing the mixture to a size smaller than 60 meshes to obtain a first mixed raw material; (2) placing the first mixed raw material at a low temperature of -28°C for 3 minutes, then rapidly heating it to 26°C within 2 seconds, repeating this process 3 times; then pressurizing it to 19 MPa, maintaining it for 6 minutes, then releasing the pressure to normal pressure, repeating this process 2 times, and obtaining a pretreated first mixed raw material; (3) Mixing the pretreated first mixed raw material and water in a weight ratio of 1:12 to obtain a first fermentation raw material, adding composite bacteria to the first fermentation raw material, wherein the composite bacteria includes Lactobacillus acidophilus, Bifidobacterium longum and Lactobacillus helveticus, and the amount of Lactobacillus acidophilus is 10 6 CFU / mL first fermentation raw material; the dosage of Bifidobacterium longum is 10 7CFU / mL first fermentation raw material; the amount of Lactobacillus helveticus is 10 6 CFU / mL first fermentation raw material; ferment at 39°C for 9h, sterilize, and obtain a first fermentation product; (4) The first fermentation product was filtered using 0.45 μm and 0.22 μm filter membranes respectively, and spray-dried to a water content of 2.37 wt % to obtain a firming composition.

[0037] The preparation method of the anti-wrinkle composition comprises the following steps: (1) mixing desert verbena leaves, polygonatum cyrtonema and sword bean seeds in a weight ratio of 1:1.4:0.6, and crushing the mixture to less than 60 meshes to obtain a second mixed raw material; (2) Mixing the second mixed raw material and water in a weight ratio of 1:9 to obtain a second fermentation raw material; adding alkaline protease to the second fermentation raw material in an amount of 3500u / mL, performing enzymatic hydrolysis at 39°C for 35 minutes, inactivating the enzyme, continuing to add papain in an amount of 2500u / mL, performing enzymatic hydrolysis at 39°C for 85 minutes, inactivating the enzyme, and finally adding neutral protease in an amount of 4500u / mL, performing enzymatic hydrolysis at 39°C for 55 minutes, inactivating the enzyme, and cooling to room temperature to obtain a pretreated second fermentation raw material; (3) Add animal Bifidobacterium to the pretreated second fermentation raw material, the amount of animal Bifidobacterium is 10 6 CFU / mL second fermentation raw material; ferment at 39°C for 6h, sterilize, and continue to add Lactobacillus casei, the amount of Lactobacillus casei is 10 9 CFU / mL of the second fermentation raw material, fermented at 39°C for 4h, sterilized, and finally added Bifidobacterium bifidum, the amount of Bifidobacterium bifidum was 10 8 CFU / mL second fermentation raw material, fermented at 39℃ for 3h, sterilized, and obtained the second fermentation product, (4) The second fermentation product was filtered using 0.45 μm and 0.22 μm filter membranes respectively, and spray-dried to a water content of 3.05% to obtain an anti-wrinkle composition.

[0038] The preparation method of the multi-type recombinant collagen firming and anti-wrinkle composition comprises the following steps: Step 1: Clean and disinfect equipment.

[0039] Step 2: Add water to the emulsifying pot, stir at 120 rpm, heat to 80°C for 40 minutes, and then cool naturally at room temperature; Step 3: Stop cooling when the temperature drops to 60°C; add a mixture of 1,3-butanediol and p-hydroxyacetophenone preheated to 60°C, stir for 15 minutes to mix evenly, and then start cooling naturally at room temperature; Step 4: Cool the room temperature naturally to 55°C, add inositol, and stir for 10 minutes to mix evenly; Step 5: Cool the room temperature naturally to 48°C, add carnosine, and stir for 10 minutes to mix evenly; Step 6: Cool the room temperature naturally to 42°C, add collagen, and stir for 10 minutes to mix evenly; Step 7: Cool the room temperature naturally to 40°C, add the remaining components, and stir for 10 minutes to mix evenly; Step 8: The product is naturally cooled to room temperature and tested for microorganisms before being bottled. It is then tested for microorganisms again before being deemed a finished product.

[0040] Example 2 The present embodiment provides a firming and anti-wrinkle composition of polytype recombinant collagen, comprising the following raw materials in weight percentage: 4% inositol, 0.1% carnosine, 0.1% collagen, 0.04% citrus peel extract, 0.5% firming composition, 0.3% anti-wrinkle composition, 0.05% 1,3-butanediol, 0.002% p-hydroxyacetophenone, and the balance is water.

[0041] The collagen comprises a mixture of recombinant type I humanized collagen, recombinant type III humanized collagen and recombinant type XVII humanized collagen in a weight ratio of 1:1.3:0.8.

[0042] The preparation method of the firming composition comprises the following steps: (1) mixing the root of Paeonia lactiflora, the bark of Pseudostellaria baicalensis, and the large-fruited passion fruit in a weight ratio of 1:0.4:1.5, and crushing the mixture to a size smaller than 60 meshes to obtain a first mixed raw material; (2) placing the first mixed raw material at a low temperature of -28°C for 3 minutes, then rapidly heating it to 25°C within 2 seconds, repeating this process twice; then pressurizing it to 20 MPa, maintaining it for 5 minutes, then releasing the pressure to normal pressure, repeating this process three times, and obtaining a pretreated first mixed raw material; (3) Mixing the pretreated first mixed raw material and water in a weight ratio of 1:15 to obtain a first fermentation raw material, adding composite bacteria to the first fermentation raw material, wherein the composite bacteria includes Lactobacillus acidophilus, Bifidobacterium longum and Lactobacillus helveticus, and the amount of Lactobacillus acidophilus is 10 6 CFU / mL first fermentation raw material; the dosage of Bifidobacterium longum is 10 7 CFU / mL first fermentation raw material; the amount of Lactobacillus helveticus is 110 7 CFU / mL first fermentation raw material; ferment at 38°C for 10 hours, sterilize, and obtain a first fermentation product; (4) The first fermentation product was filtered using 0.45 μm and 0.22 μm filter membranes respectively, and spray-dried to a water content of 2.53 wt % to obtain a firming composition.

[0043] The preparation method of the anti-wrinkle composition comprises the following steps: (1) mixing desert verbena leaves, polygonatum cyrtonema and sword bean seeds in a weight ratio of 1:1.5:0.7, and crushing the mixture to less than 60 meshes to obtain a second mixed raw material; (2) Mixing the second mixed raw material and water at a weight ratio of 1:10 to obtain a second fermentation raw material; adding alkaline protease to the second fermentation raw material in an amount of 4000u / mL of the second fermentation raw material, performing enzymatic hydrolysis at 40°C for 30 minutes, inactivating the enzyme, continuing to add papain in an amount of 3000u / mL of the second fermentation raw material, continuing to perform enzymatic hydrolysis at 38°C for 90 minutes, inactivating the enzyme, and finally adding neutral protease in an amount of 4000u / mL of the second fermentation raw material, performing enzymatic hydrolysis at 40°C for 50 minutes, inactivating the enzyme, and cooling to room temperature to obtain a pretreated second fermentation raw material; (3) Add animal Bifidobacterium to the pretreated second fermentation raw material, the amount of animal Bifidobacterium is 10 6 CFU / mL second fermentation raw material; ferment at 40℃ for 5h, sterilize, and continue to add Lactobacillus casei, the amount of Lactobacillus casei is 10 9 CFU / mL of the second fermentation raw material, fermented at 38°C for 5h, sterilized, and finally added Bifidobacterium bifidum, the amount of Bifidobacterium bifidum was 10 7 CFU / mL second fermentation raw material, fermented at 40℃ for 2h, sterilized, and obtained the second fermentation product, (4) The second fermentation product was filtered using 0.45 μm and 0.22 μm filter membranes respectively, and spray-dried to a water content of 2.97 wt % to obtain an anti-wrinkle composition.

[0044] The preparation method of the multi-type recombinant collagen firming and anti-wrinkle composition comprises the following steps: Step 1: Clean and disinfect equipment.

[0045] Step 2: Add water to the emulsifier, stir at 120 rpm, heat to 85°C for 30 minutes, and then start cooling; Step 3: When the room temperature naturally drops to 60°C, stop cooling; add a mixture of 1,3-butanediol and p-hydroxyacetophenone preheated to 65°C, stir for 10 minutes to mix evenly, and then start cooling; Step 4: Cool the room temperature naturally to 54°C, add inositol, and stir for 10 minutes to mix evenly; Step 5: Cool the room temperature naturally to 50°C, add carnosine, and stir for 10 minutes to mix evenly; Step 6: Cool the room temperature naturally to 42°C, add collagen, and stir for 10 minutes to mix evenly; Step 7: Cool the room temperature naturally to 38°C, add the remaining components, and stir for 10 minutes to mix evenly; Step 8: Cool naturally to room temperature, test for microorganisms and then bottle it. Then test for microorganisms again and it is the finished product.

[0046] Comparative Example 1 The difference between this comparative example and Example 1 is: a firming and anti-wrinkle composition of polytype recombinant collagen, comprising the following raw materials in weight percentage: 0.1% inositol, 1.2% carnosine, 0.19% collagen, 0.01% citrus peel extract, 0.34% firming composition, 0.43% anti-wrinkle composition, 0.04% 1,3-butanediol, 0.006% p-hydroxyacetophenone, and the balance is water.

[0047] Comparative Example 2 The difference between this comparative example and Example 1 is that the root of Paeonia lactiflora, the bark of Pseudostellaria baicalensis, and the large-fruited passion fruit are mixed in a weight ratio of 1:1:1, and then crushed to less than 60 meshes after mixing to obtain a first mixed raw material.

[0048] Comparative Example 3 The difference between this comparative example and Example 1 is that the preparation method of the firming composition comprises the following steps: (1) mixing the root of Paeonia lactiflora, the bark of Pseudostellaria baicalensis, and the large-fruited passion fruit in a weight ratio of 1:0.4:1.6, and crushing the mixture to a size smaller than 60 meshes to obtain a first mixed raw material; (2) Mixing the first mixed raw material and water in a weight ratio of 1:12 to obtain a first fermentation raw material, adding composite bacteria to the first fermentation raw material, wherein the composite bacteria includes Lactobacillus acidophilus, Bifidobacterium longum and Lactobacillus helveticus, and the amount of Lactobacillus acidophilus is 10 6 CFU / mL first fermentation raw material; the dosage of Bifidobacterium longum is 10 7 CFU / mL first fermentation raw material; the amount of Lactobacillus helveticus is 10 6 CFU / mL first fermentation raw material; ferment at 39°C for 9h, sterilize, and obtain a first fermentation product; (3) The first fermentation product was filtered using 0.45 μm and 0.22 μm filter membranes respectively, and spray-dried to a water content of 2.37 wt % to obtain a firming composition.

[0049] Comparative Example 4 The difference between this comparative example and Example 1 is that the preparation method of the firming composition comprises the following steps: (1) mixing the root of Paeonia lactiflora, the bark of Pseudostellaria baicalensis, and the large-fruited passion fruit in a weight ratio of 1:0.4:1.6, and crushing the mixture to a size smaller than 60 meshes to obtain a first mixed raw material; (2) placing the first mixed raw material at a low temperature of -28°C for 3 minutes, then rapidly heating it to 26°C within 2 seconds, repeating this process 3 times; then pressurizing it to 19 MPa, maintaining it for 6 minutes, then releasing the pressure to normal pressure, repeating this process 2 times, and obtaining a pretreated first mixed raw material; (3) Mixing the pretreated first mixed raw material and water in a weight ratio of 1:12 to obtain a first fermentation raw material, adding composite bacteria to the first fermentation raw material, wherein the composite bacteria includes animal Bifidobacterium, Lactobacillus casei and Bifidobacterium bifidum, and the amount of animal Bifidobacterium is 10 6 CFU / mL first fermentation raw material; the dosage of Lactobacillus casei is 10 7 CFU / mL first fermentation raw material; the amount of Bifidobacterium bifidum is 10 6 CFU / mL first fermentation raw material; ferment at 39°C for 9h, sterilize, and obtain a first fermentation product; (4) The first fermentation product was filtered using 0.45 μm and 0.22 μm filter membranes respectively, and spray-dried to a water content of 2.37 wt % to obtain a firming composition.

[0050] Comparative Example 5 The difference between this comparative example and Example 1 is that desert verbena leaves, polygonatum cyrtonema and sword bean seeds in a weight ratio of 1:1:1 are mixed and crushed to less than 60 meshes to obtain a second mixed raw material.

[0051] Comparative Example 6 The difference between this comparative example and Example 1 is that different enzymes are used.

[0052] The preparation method of the anti-wrinkle composition comprises the following steps: (1) mixing desert verbena leaves, polygonatum cyrtonema and sword bean seeds in a weight ratio of 1:1.4:0.6, and crushing the mixture to less than 60 meshes to obtain a second mixed raw material; (2) Mixing the second mixed raw material and water in a weight ratio of 1:9 to obtain a second fermentation raw material; adding bromelain to the second fermentation raw material in an amount of 3500u / mL of the second fermentation raw material, performing enzymatic hydrolysis at 39°C for 35 minutes, inactivating the enzyme, continuing to add pectinase in an amount of 2500u / mL of the second fermentation raw material, continuing to perform enzymatic hydrolysis at 39°C for 85 minutes, inactivating the enzyme, and finally adding β-glucanase in an amount of 4500u / mL of the second fermentation raw material, performing enzymatic hydrolysis at 39°C for 55 minutes, inactivating the enzyme, and cooling to room temperature to obtain a pretreated second fermentation raw material; (3) Add animal Bifidobacterium to the pretreated second fermentation raw material, the amount of animal Bifidobacterium is 10 6 CFU / mL second fermentation raw material; ferment at 39°C for 6h, sterilize, and continue to add Lactobacillus casei, the amount of Lactobacillus casei is 10 9CFU / mL of the second fermentation raw material, fermented at 39°C for 4h, sterilized, and finally added Bifidobacterium bifidum, the amount of Bifidobacterium bifidum was 10 8 CFU / mL second fermentation raw material, fermented at 39℃ for 3h, sterilized, and obtained the second fermentation product, (4) The second fermentation product was filtered using 0.45 μm and 0.22 μm filter membranes respectively, and spray-dried to a water content of 3.05 wt % to obtain an anti-wrinkle composition.

[0053] Comparative Example 7 The difference between this comparative example and Example 1 is that different bacteria are used.

[0054] The preparation method of the anti-wrinkle composition comprises the following steps: (1) mixing desert verbena leaves, polygonatum cyrtonema and sword bean seeds in a weight ratio of 1:1.4:0.6, and crushing the mixture to less than 60 meshes to obtain a second mixed raw material; (2) Mixing the second mixed raw material and water in a weight ratio of 1:9 to obtain a second fermentation raw material; adding alkaline protease to the second fermentation raw material in an amount of 3500u / mL, performing enzymatic hydrolysis at 39°C for 35 minutes, inactivating the enzyme, continuing to add papain in an amount of 2500u / mL, performing enzymatic hydrolysis at 39°C for 85 minutes, inactivating the enzyme, and finally adding neutral protease in an amount of 4500u / mL, performing enzymatic hydrolysis at 39°C for 55 minutes, inactivating the enzyme, and cooling to room temperature to obtain a pretreated second fermentation raw material; (3) Add Lactobacillus acidophilus to the pretreated second fermentation raw material, and the amount of Lactobacillus acidophilus is 10 6 CFU / mL second fermentation raw material; ferment at 39℃ for 6h, sterilize, and continue to add Lactobacillus helveticus, the amount of Lactobacillus helveticus is 10 9 CFU / mL of the second fermentation raw material, fermented at 39°C for 4h, sterilized, and finally added Bifidobacterium longum. The amount of Bifidobacterium longum was 10 8 CFU / mL second fermentation raw material, fermented at 39℃ for 3h, sterilized, and obtained the second fermentation product, (4) The second fermentation product was filtered using 0.45 μm and 0.22 μm filter membranes respectively, and spray-dried to a water content of 3.05 wt % to obtain an anti-wrinkle composition.

[0055] Comparative Example 8 The difference between this comparative example and Example 1 is that the collagen comprises a mixture of recombinant type I humanized collagen, recombinant type III humanized collagen and recombinant type XVII humanized collagen in a weight ratio of 1:1:1.

[0056] Performance Testing The firming and anti-wrinkle compositions prepared in Examples 1-2 and Comparative Examples 1-8 were used as samples for performance testing.

[0057] 1. Human skin patch test Test population: Volunteers aged between 20 and 50 were selected and randomly divided into 10 groups, with 20 people in each group; Test method: First clean the skin of the test area with physiological saline and wait for 5 minutes to dry; select a qualified patch tester, take 0.5mL of sample into the patch tester in a closed patch test, and apply special tape to the back of the subject, and ask the subject how it feels; bathing is prohibited during the test; the subject keeps the patch area dry within 48 hours, avoids intense exercise, scratching the patch area, and long-term sunlight exposure, etc. After 48 hours, remove the tester and mark it. After 30 minutes, wait for the indentation to disappear and make a judgment under sufficient light. Record the results according to the skin reaction grade standard in the "Hygiene Standards for Skin Care Products". The skin adverse reaction grade standard is shown in Table 1 below. If there is any discomfort during the process, stop the test immediately. The results are shown in Table 2.

[0058] Table 1 Grading of skin adverse reactions

[0059] Table 2 Human skin patch test results

[0060] It can be seen from Table 2 that the firming and anti-wrinkle composition prepared by the present invention is mild and non-irritating.

[0061] 2. Skin firming test Randomly select 20-50-year-old test subjects and divide them into 10 groups, 10 in each group. Use the firming and anti-wrinkle composition once in the morning and evening, evenly apply it on the whole face according to the size of the face, for 30 days. During the test, the subjects are not allowed to use other items to maintain skin elasticity and cosmetics that affect the results, and do not change their original eating habits during the test.

[0062] Cutometer MPA580 was used to test the skin condition on the 0th day, 30th day and 6th month. The test site was the zygomatic area. The probe was used to measure the same test area 3 times and the average value was taken. The test parameter was R2 (the ratio of the skin's rebound amount Ua when there was no negative pressure to the maximum stretch amount Uf when there was negative pressure). The closer R2 is to 1, the better the skin's elasticity. The average change value of R2 on the 30th day = the R2 value on the 30th day - the R2 value on the 0th day, the average change rate of R2 = the average change value of R2 on the 30th day / the R2 value on the 0th day.

[0063] The average change value of R2 in the 6th month = R2 value in the 6th month - R2 value on the 0th day, the average change rate of R2 in the 6th month = average change value of R2 in the 6th month / R2 value on the 0th day.

[0064] The specific test results of skin elasticity are shown in Table 3.

[0065] Table 3 Short-term and long-term skin elasticity test results Average change of R2 on day 30 % Average change of R2 in the 6th month (%) Example 1 15.54 54.64 Example 2 15.12 53.18 Comparative Example 1 11.51 44.93 Comparative Example 2 12.40 47.25 Comparative Example 3 13.76 46.47 Comparative Example 4 12.23 45.72 Comparative Example 5 13.85 48.99 Comparative Example 6 12.94 46.23 Comparative Example 7 13.48 47.38 Comparative Example 8 14.63 50.01 It can be seen from Table 3 that the effect of the anti-wrinkle and firming composition decreases when the preparation method of the firming composition is changed. At the same time, the preparation method of the anti-wrinkle composition and the composition of collagen also affect the firming effect.

[0066] 3. Anti-wrinkle test Testers: 100 volunteers, aged 23-60 years old; the volunteers were divided into 10 groups, 10 in each group, so that the initial skin wrinkle conditions in each group were as similar as possible, and the firming and anti-wrinkle composition was tested separately.

[0067] Inclusion criteria: volunteers with skin problems such as wrinkles, but no history of skin and systemic diseases, and no use of hormone drugs in the past month; Product use rules: 2-3 days before the test and during the entire test period, do not change skin care products, once a day in the morning and once in the evening. Select the forehead skin as the test area, apply the firming anti-wrinkle composition to the left half of each person's forehead, and do not apply it to the right half, observe the skin condition of the test area, continue to use this product, and observe for 3 consecutive months. Compare the improvement of wrinkles before use and after 7 days and 90 days of use, measure each area 3 times each time, calculate the average value, and then calculate the average value within the group.

[0068] Evaluation criteria: 10 points, no wrinkles; 7-9 points, 1-2 wrinkles, less than 1.5cm in length; 4-6 points, 3-5 wrinkles, less than 3cm in length; 1-3 points: there are several main wrinkles, and the length is greater than 3cm.

[0069] The final scores are shown in Table 4 below.

[0070] Table 4 Anti-wrinkle performance test results

[0071] It can be seen from Table 4 that the anti-wrinkle effect of the anti-wrinkle and firming composition decreases when the preparation method of the anti-wrinkle composition is changed. At the same time, the preparation method of the anti-wrinkle composition and the composition of collagen also have an impact on the anti-wrinkle effect.

[0072] 3. Whitening effect: 100 female volunteers (25-45 years old) with sensitive skin were divided into 10 groups, each with 10 people, and used the firming and anti-wrinkle composition respectively.

[0073] Test area: face (blank sample on the left, sample on the right).

[0074] Use twice a day, morning and evening, for 4 weeks.

[0075] Testing instrument: Finnish Delfin skin tester, model: DelfinSkinColorCatch.

[0076] Effect index: Determine the change in color brightness (ΔL*). The calculation formula is: ΔL* = [L* (4 weeks after the comparative example or embodiment is tested) L* (before the test)] [L* (1 month after the blank example is tested) L* (before the blank example is tested)], and calculate the average ΔL* of each group. The test results are shown in Table 5.

[0077] Table 5 Whitening test results ΔL*Average Example 1 5.12 Example 2 5.06 Comparative Example 1 3.88 Comparative Example 2 4.61 Comparative Example 3 4.44 Comparative Example 4 4.38 Comparative Example 5 4.72 Comparative Example 6 4.39 Comparative Example 7 4.57 Comparative Example 8 4.86 As shown in Table 5, the changes in the compositions of the firming composition, the anti-wrinkle composition and the collagen lead to a decrease in the whitening effect of the firming and anti-wrinkle composition.

[0078] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A firming and anti-wrinkle composition of polytype recombinant collagen, characterized in that: The invention comprises the following raw materials in percentage by weight: 0.5-5% inositol, 0.1-0.5% carnosine, 0.02-0.15% collagen, 0.003-0.15% citrus peel extract, 0.4-0.7% firming composition, 0.1-0.4% anti-wrinkle composition, 0.01-0.05% butylene glycol, 0.002-0.009% p-hydroxyacetophenone, and the balance is water; The preparation method of the firming composition comprises the following steps: (1) Mixing the root of Paeonia lactiflora, the bark of the wild sycamore tree, and the large-fruited passion fruit, and crushing the mixture to a size smaller than 60 meshes to obtain a first mixed raw material; (2) placing the first mixed raw material at a low temperature of -28°C for 2-3 minutes, then rapidly heating it to 25-27°C within 2 seconds, repeating this process 2-3 times; then pressurizing it to 18-20 MPa, maintaining it for 5-6 minutes, then releasing the pressure to normal pressure, repeating this process 2-3 times, and obtaining a pretreated first mixed raw material; (3) Mixing the pretreated first mixed raw material and water in a weight ratio of 1: (10-15) to obtain a first fermentation raw material, adding composite bacteria to the first fermentation raw material, wherein the composite bacteria includes Lactobacillus acidophilus, Bifidobacterium longum and Lactobacillus helveticus, and the amount of Lactobacillus acidophilus is 10 5 -10 6 CFU / mL first fermentation raw material; the dosage of Bifidobacterium longum is 10 7 -10 8 CFU / mL first fermentation raw material; the amount of Lactobacillus helveticus is 10 6 -10 7 CFU / mL first fermentation raw material; ferment at 38-40°C for 8-10h, sterilize, and obtain a first fermentation product; (4) Filtering the first fermentation product, spray-drying it, and obtaining a firming composition.

2. The firming and anti-wrinkle composition of multi-type recombinant collagen according to claim 1, characterized in that: The weight ratio of Sichuan red peony root, wild sycamore bark, and passion fruit is 1: (0.3-0.5): (1.5-1.7).

3. The firming and anti-wrinkle composition of multi-type recombinant collagen according to claim 1, characterized in that: The preparation method of the anti-wrinkle composition comprises the following steps: (1) mixing desert verbena leaves, polygonatum cyrtonema and sword bean seeds, and crushing them into a size smaller than 60 meshes to obtain a second mixed raw material; (2) Mixing the second mixed raw material and water in a weight ratio of 1: (8-10) to obtain a second fermentation raw material; adding alkaline protease in an amount of 3000-4000u / mL of the second fermentation raw material, performing enzymatic hydrolysis at 38-40°C for 30-40min, inactivating the enzyme, continuing to add papain in an amount of 2000-3000u / mL of the second fermentation raw material, continuing to perform enzymatic hydrolysis at 38-40°C for 80-90min, inactivating the enzyme, and finally adding neutral protease in an amount of 4000-5000u / mL of the second fermentation raw material, performing enzymatic hydrolysis at 38-40°C for 50-60min, inactivating the enzyme, and cooling to room temperature to obtain a pretreated second fermentation raw material; (3) Add animal Bifidobacterium to the pretreated second fermentation raw material, the amount of animal Bifidobacterium is 10 6 -10 7 CFU / mL second fermentation raw material; ferment at 38-40℃ for 5-7h, sterilize, and continue to add Lactobacillus casei, the amount of Lactobacillus casei is 10 8 -10 9 CFU / mL second fermentation raw material, ferment at 38-40℃ for 3-5h, sterilize, and finally add Bifidobacterium bifidum, the dosage of Bifidobacterium bifidum is 10 7 -10 8 CFU / mL second fermentation raw material, ferment at 38-40℃ for 2-4h, sterilize to obtain the second fermentation product, (4) filtering the second fermentation product, spray-drying it, and obtaining an anti-wrinkle composition.

4. The firming and anti-wrinkle composition of multi-type recombinant collagen according to claim 3, characterized in that: The weight ratio of the desert verbena leaves, polygonatum cyrtonema and sword bean seeds is 1: (1.3-1.5): (0.5-0.7).

5. The firming and anti-wrinkle composition of multi-type recombinant collagen according to claim 1, characterized in that: The collagen is selected from one or more of recombinant type I humanized collagen, recombinant type III humanized collagen, and recombinant type XVII humanized collagen.

6. The firming and anti-wrinkle composition of multi-type recombinant collagen according to claim 5, characterized in that: The collagen comprises a mixture of recombinant type I humanized collagen, recombinant type III humanized collagen and recombinant type XVII humanized collagen in a weight ratio of 1: (1.3-1.5): (0.6-0.8).

7. The firming and anti-wrinkle composition of multi-type recombinant collagen according to claim 1, characterized in that: The weight ratio of collagen, firming composition and anti-wrinkle composition is (0.8-1):(5-6):(2-3).

8. The firming and anti-wrinkle composition of multi-type recombinant collagen according to claim 1, characterized in that: The invention comprises the following raw materials in percentage by weight: 1% inositol, 0.3% carnosine, 0.09% collagen, 0.11% citrus peel extract, 0.54% firming composition, 0.23% anti-wrinkle composition, 0.04% butylene glycol, 0.006% p-hydroxyacetophenone, and the balance is water.

9. The firming and anti-wrinkle composition of multi-type recombinant collagen according to claim 1, characterized in that: The preparation method of the firming and anti-wrinkle composition of multi-type recombinant collagen comprises the following steps: Step 1: Clean and disinfect equipment; Step 2: Add water to the emulsifier, start stirring, heat to 80-85℃ and keep warm for 30-40 minutes, then start cooling down; Step 3: Stop cooling when the temperature drops to 60°C; add a mixture of butanediol and p-hydroxyacetophenone preheated to 60-65°C, stir until uniform, and then start cooling; Step 4: Cool down, add inositol, and stir until uniform; Step 5: Cool down, add carnosine, and stir until uniform; Step 6: Cool down, add collagen, and stir until uniform; Step 7: Cool down, add remaining ingredients, and stir until uniform; Step 8: After cooling and passing the microbiological test, the product is bottled and then tested for microorganisms again to be the finished product.

10. Use of the firming and anti-wrinkle composition of multi-type recombinant collagen according to any one of claims 1 to 9 in the preparation of cosmetics.

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