Anti-aging cosmetics containing small molecule plant fermentation products and preparation method thereof

Abstract

The present invention relates to the field of cosmetic technology, and in particular to an anti-aging cosmetic containing a small molecule plant fermentation product and a preparation method thereof. The anti-aging cosmetic includes 0.5%-5% of a small molecule plant fermentation product, 10%-20% of a moisturizer, 0.5%-1.5% of sodium chloride or magnesium sulfate, 0.5%-2% of a thickener, 0.1%-2% of a skin conditioner, 3%-5% of an emulsifier, 5%-35% of an emollient, 0.01%-0.5% of a fragrance, 0.1%-3% of a filler, and a water balance; the small molecule plant fermentation product includes a secondary metabolite obtained by fermenting a composite lactobacillus with a polygonatum root extract, a tuckahoe sclerotium extract, an avocado fruit extract, and a ginseng root extract as a composite plant raw material. The cosmetics of the present invention have excellent anti-aging effects, high stability, and a good skin feel experience.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and in particular to an anti-aging cosmetic containing a small-molecule plant fermentation product and a preparation method thereof. Background Art

[0002] Skin aging refers to the process of skin's external morphology, structure, and function decline under the influence of exogenous or endogenous factors. From the perspective of internal factors, the natural aging of cells gradually slows down the metabolism rate and prolongs the cell renewal cycle. Among the external factors, ultraviolet rays are one of the key factors leading to skin aging. Long-term exposure to ultraviolet rays will destroy the collagen and elastic fibers in the skin, thereby accelerating the formation of wrinkles. Environmental pollution, smoking, and bad eating habits can also cause damage to skin health. The endogenous and exogenous aging of the skin are interrelated and affect each other. The clinical manifestations of skin aging are dry, rough, thin, reduced elasticity, loss of luster, sagging, atrophy, wrinkles of varying degrees, pigmentation, capillary dilation, and the disorder of the skin's physiological structure and functional degradation.

[0003] Chinese invention patent CN114306112B provides an anti-aging composition and a preparation method thereof, wherein the anti-aging composition comprises the following components: dipotassium glycyrrhizinate, sodium hyaluronate, sucrose stearate, vitamin E, fucoidan, polygonatum extract, saffron extract, glycerol, 1,3-butylene glycol and deionized water, which can achieve good anti-aging, moisturizing and anti-inflammatory effects and improve anti-aging effects. Chinese invention patent CN107802540B discloses an anti-aging composition, which comprises a polypeptide component and an anti-aging plant extract, which can play a synergistic role and achieve better anti-aging effects.

[0004] However, the above anti-aging composition cannot delay aging comprehensively from multiple pathways, and it also needs to be combined with substances of other action pathways to better play the anti-wrinkle and firming effect.

[0005] In addition, oil-in-water and water-in-oil are two main dosage forms of skin care products, which have significant differences in ingredient composition, texture characteristics, usage experience and skin care efficacy.

[0006] Oil-in-water is a structure in which oil molecules enclose water molecules. The main component of this dosage form is oil, and its outer layer is composed of an oil film, so the texture is usually greasy, thick, and has an opaque appearance. Due to the skin-friendly nature of the outer oil film, this dosage form can blend well with the stratum corneum of the human skin, thereby achieving a deep hydration effect.

[0007] Oil-in-water is a structure where water molecules wrap around oil molecules. Its main component is water, so its texture is relatively refreshing and is usually presented in the form of an emulsion. The surfactants in the water-in-water formulation are hydrophilic, which allows water to quickly penetrate the skin after being applied to the skin, while the oil can also be quickly absorbed by the skin, allowing the skin care products to perform better.

[0008] How to choose the right product dosage form and cosmetic excipients so that the product has excellent anti-aging effects, stability and skin feel is a major challenge that needs to be overcome.

[0009] Therefore, it is very necessary to develop an anti-aging cosmetic and a preparation method thereof that can solve the above-mentioned technical problems. Summary of the invention

[0010] The purpose of the present invention is to overcome the deficiencies of the prior art and provide an anti-aging cosmetic containing small molecule plant fermentation products with excellent anti-aging effect, high stability and good skin feel, and a preparation method thereof.

[0011] The present invention is achieved through the following technical solutions:

[0012] An anti-aging cosmetic containing small molecule plant fermentation products, comprising the following components in terms of mass percentage:

[0013] Oil phase:

[0014] Thickener 0.5%-2%;

[0015] Emulsifier 3%-5%;

[0016] Emollient 5%-35%;

[0017] Filler 0.1%-3%;

[0018] Aqueous phase:

[0019] Small molecule plant fermentation 0.5%-5%;

[0020] Moisturizer 10%-20%;

[0021] Sodium chloride or magnesium sulfate 0.5%-1.5%;

[0022] Skin conditioner 0.1%-2%;

[0023] Fragrance 0.01%-0.5%;

[0024] Water balance;

[0025] The pH of the aqueous phase is controlled within the range of 5.0-7.0;

[0026] The small molecule plant fermentation product comprises, by weight: 95-99 parts of composite plant fermentation concentrate and 1-5 parts of protective agent;

[0027] The composite plant fermentation concentrate uses polygonatum root extract, tuckahoe sclerotium extract, avocado fruit extract, and ginseng root extract as composite plant raw materials, and is fermented with composite lactobacillus to obtain secondary metabolites;

[0028] The composite lactobacillus is composed of at least two of plant lactobacillus, fermented mucus lactobacillus, casei lactobacillus and acidophilus lactobacillus.

[0029] In some embodiments of the present invention, the plant lactobacillus is plant lactobacillus 14917, the fermented mucus lactobacillus is fermented mucus lactobacillus CICC 25124, the casei lactobacillus is human casei lactobacillus NCU011061, and the acidophilus lactobacillus is acidophilus NCFM.

[0030] The secondary metabolites obtained by complex lactobacillus fermentation are also called "lactobacillus fermentation lysate", which is the product of inactivation and filtration after lactobacillus fermentation. Lactobacillus fermentation lysate is rich in lactic acid, peptidoglycan, teichoic acid, protein, phospholipids, sterols, fatty acids, peptides and amino acids, nucleotides, extracellular polysaccharides, and multiple vitamins, and can be used for skin conditioning, anti-aging, antibacterial, anti-inflammatory, moisturizing, etc.

[0031] The composite plant fermentation concentrate used in the present invention is a brown liquid with a unique fermentation smell. Due to the low pH and strong ionicity of the raw material, when used in the preparation of skin care products, it is difficult to thicken, has a poor skin feel, and has a strong odor.

[0032] In some embodiments of the present invention, the protective agent includes: one or more of 1,2-hexanediol, 1,2-pentanediol, caprylyl glycol, and ethylhexylglycerol.

[0033] In some embodiments of the present invention, the composite plant fermentation concentrate is prepared from the following raw materials in weight percentage: 4%-11% polygonatum root extract, 2%-9% tuckahoe sclerotium extract, 1%-6% avocado fruit extract, 1%-6% ginseng root extract, 2%-8% composite lactobacillus and the remainder water.

[0034] In some embodiments of the present invention, the preparation method of the small molecule plant fermentation product includes: mixing polygonatum root extract, tuckahoe sclerotium extract, avocado fruit extract, ginseng root extract and water, adding the expanded composite lactobacillus, filtering to obtain a filtrate after fermentation, adding a protective agent, and stirring evenly to obtain the product.

[0035] Specifically, the fermentation condition is 35-39°C fermentation.

[0036] Preferably, the fermentation condition is 37°C.

[0037] Specifically, the fermentation time is 5-10 days.

[0038] Preferably, the fermentation time is 7 days.

[0039] Specifically, the filtration is carried out using a ceramic membrane.

[0040] Preferably, the filtration is carried out using a 0.1 μm plate and frame filter.

[0041] In some embodiments of the present invention, the moisturizer includes one or more of glycerol, butylene glycol, 1,2-hexanediol, propylene glycol, 1,2-pentanediol, ethylhexylglycerol and HTsoft_5688. Among them, the main components of HTsoft_5688 are: dipropylene glycol, p-hydroxyacetophenone, 1,2-hexanediol, caprylyl glycol and isopentyl glycol.

[0042] In some embodiments of the present invention, the pH adjuster used to adjust the pH of the aqueous phase includes at least one of sodium hydroxide, citric acid, aminomethyl propanol and triethanolamine.

[0043] In some embodiments of the present invention, the thickener is quaternary ammonium salt-18 bentonite.

[0044] In some embodiments of the present invention, the skin conditioning agent includes one or more of glycerophosphoinositol choline salt, palmitoyl tripeptide-5, hydrolyzed collagen, ferulic acid, ergothioneine, acetyl tetrapeptide-2, hydroxypropyl tetrahydropyrantriol, tetrahydromethylpyrimidine carboxylic acid, carbohydrate isomers, parahydroxyacetophenone, ceramide NP, sodium hyaluronate, allantoin, dipotassium glycyrrhizinate, acetyl hexapeptide-8, acetylated sodium hyaluronate and soluble collagen.

[0045] In some embodiments of the present invention, the emulsifier includes one or more of Winsier Plus (PEG-10 dimethicone, phenyl dimethicone, disteardimonium hectorite), lauryl PEG-8 dimethicone, and cetyl PEG / PPG-10 / 1 dimethicone.

[0046] In some preferred embodiments of the present invention, the emulsifier includes Winsier Plus, cetyl PEG / PPG-10 / 1 dimethicone and lauryl PEG-8 dimethicone.

[0047] In some more preferred embodiments of the present invention, the mass ratio of Winsier Plus, cetyl PEG / PPG-10 / 1 dimethicone and lauryl PEG-8 dimethicone is 1-6:1:1.

[0048] In some embodiments of the present invention, the emollient includes silicone oil emollient and grease emollient.

[0049] In some preferred embodiments of the present invention, the silicone oil emollient includes at least one of polydimethylsiloxane and cyclopentasiloxane.

[0050] In some preferred embodiments of the present invention, the oil-based emollient includes at least one of ethylhexyl palmitate, isohexadecane, caprylic / capric triglyceride, glyceryl tri(ethylhexanoate), hydrogenated polydecene and neopentyl glycol dicaprylate / dicaprate.

[0051] In some embodiments of the present invention, the fragrance comprises at least one of essential oil and essence.

[0052] In some embodiments of the present invention, the filler is silica.

[0053] The present invention also relates to a method for preparing the above-mentioned anti-aging cosmetics, comprising the following steps:

[0054] (1) Add water and moisturizer to a water pot, heat up, stir until completely dissolved, cool down, add small molecule plant fermentation, sodium chloride or magnesium sulfate, and skin conditioner, stir until completely dissolved, measure the pH of the material, adjust the pH of the material to 5.0-7.0, stir until completely dissolved, and obtain an aqueous phase;

[0055] (2) Add thickener, emulsifier, emollient and filler into the emulsifying pot, stir and homogenize under vacuum conditions, add water phase after the material is uniform, stir and homogenize under vacuum conditions, then add fragrance and stir evenly.

[0056] In some embodiments of the present invention, the temperature of the heating in step (1) is 80-90°C, and the heating is maintained for 30-60 minutes; the temperature of the cooling is below 38°C.

[0057] In some embodiments of the present invention, the vacuum condition in step (2) is -0.04 MPa to -0.06 MPa.

[0058] The beneficial effects of the present invention are:

[0059] The small molecule plant fermentation product of the present invention is prepared by fermenting a variety of plant extracts, and comprehensively improves skin wrinkles by scavenging free radicals, promoting type I collagen and elastin regeneration, and resisting glycation, and has a good anti-wrinkle and firming effect.

[0060] The present invention is designed as an oil-in-water system formula, which achieves the purpose of compounding a high content of small molecule plant fermentation products, and also has the product characteristics of less odor, light, non-sticky, and easy to absorb. The application of oil-in-water system design has the following advantages: ① The water-soluble composite plant fermentation concentrate exists in the inner layer of the oil droplet molecule. Due to the oil wrapping of the outer layer, the contact with the outer layer oxygen is reduced, and the oxidation fermentation rate of the inner layer composite plant fermentation concentrate slows down, and the product taste is relatively small. The fermentation taste can be masked by compounding essential oils / essences, so that consumers can enjoy a pleasant smell; ② The water-soluble composite plant fermentation concentrate exists in the inner layer of the oil droplet molecule, and does not require ion-resistant emulsifiers and thickeners. By selecting and proportioning oil-in-water emulsifiers, thickeners, emollients, etc., the ideal consistency can be achieved, and the skin feel can be made light and easy to absorb.

[0061] The present invention improves the stability of the product while ensuring the anti-aging effect by screening the amount of small molecule plant fermentation products used in the cosmetics.

[0062] The invention screens the auxiliary material composition, compounds the specific emulsifier and thickener, realizes the thickening of low pH and high ionic raw materials, and improves the product stability.

[0063] The invention improves the anti-aging effect and soothing effect of the cosmetics by adding a specific skin conditioning agent.

[0064] The invention adds a specific filler to retain the moisturizing feeling of the oil film formed on the skin surface while reducing the stickiness of the oil film, thereby improving the skin feel of the product and giving the product the characteristics of being light, non-sticky and easily absorbed. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] Figure 1 This is a bar graph of the anti-glycation experiment of Experimental Example 1; in the figure, A is the anti-glycation effect of a small molecule plant fermentation product with a concentration of 2.5%; B is the anti-glycation effect of a small molecule plant fermentation product with a concentration of 5%.

[0066] Figure 2 This is the scavenging effect diagram of DPPH free radicals in Experimental Example 2.

[0067] Figure 3 This is the scavenging effect diagram of ABTS free radicals in Experimental Example 2.

[0068] Figure 4 This is a bar graph of the relative expression of type I collagen gene in zebrafish in Experimental Example 3; * in the figure represents a significant difference (p < 0.05).

[0069] Figure 5This is a bar graph of the relative expression of zebrafish elastin gene in Experimental Example 4; * in the figure represents a significant difference (p < 0.05). DETAILED DESCRIPTION

[0070] The present invention will be further described below in conjunction with specific embodiments, and the advantages and features of the present invention will become clearer as the description proceeds. However, these embodiments are exemplary only and do not constitute any limitation to the scope of the present invention. It should be understood by those skilled in the art that the details and forms of the technical solution of the present invention may be modified or replaced without departing from the spirit and scope of the present invention, but these modifications and replacements all fall within the scope of protection of the present invention.

[0071] the term:

[0072] In the present invention, the term "half maximal clearance rate (EC50)" refers to: the sample concentration that achieves 50% clearance of DPPH free radicals; the lower the EC50, the stronger the free radical scavenging ability of the substance.

[0073] In the present invention, the term "ascorbic acid equivalent (AEAC)" refers to how much mg ascorbic acid (VC) the antioxidant capacity of 100 g of a sample is equivalent to.

[0074] In the present invention, the term "zebrafish embryo" refers to a zebrafish embryo at a developmental stage that relies on yolk for energy.

[0075] The materials used in the present invention are shown in Table 1:

[0076] Table 1 Materials used in the present invention

[0077]

[0078] Example 1 Preparation of small molecule plant fermentation products

[0079] 1. Preparation of compound lactobacillus culture solution

[0080] The composite lactobacillus culture solution contains, by weight percentage, 8% of polygonatum sibiricum extract, 6% of tuckahoe sclerotium extract, 3% of avocado fruit extract, 3% of ginseng root extract and the balance of deionized water.

[0081] The above-mentioned composite lactobacillus culture solution was added into a triangular flask, mixed evenly, a cotton plug was added to the bottle mouth, and the bottle mouth was wrapped with newspaper, and sterilized at 115° C. for 20 min in a portable high-pressure sterilizer, and then taken out for use.

[0082] 2. Expansion of bacterial strains

[0083] Under sterile conditions, use an inoculation loop to pick a loop of Lactobacillus plantarum 14917, Lactobacillus fermentans CICC25124, Lactobacillus casei NCU011061, and Lactobacillus acidophilus NCFM respectively, transfer them into different sterilized test tubes, draw a curve, and culture them in horizontal test tubes at 32°C for 7 days. After the colonies grow, transfer them again under the same operating conditions, and the culture conditions are the same as above. The strains are used in the experiment after 3 generations of activation process.

[0084] 3. Preparation of composite plant fermentation concentrate

[0085] The selected culture temperature is 37±2°C, sterile water is added to the activated slant test tube to elute the spores, the spore suspension is aspirated with a sterile pipette, and the spore suspension is transferred into the composite lactobacillus culture solution prepared in step 1 at a 5% inoculum amount (the weight ratio of Lactobacillus plantarum 14917, Lactobacillus fermentative CICC 25124, Lactobacillus casei NCU011061 and Lactobacillus acidophilus NCFM is 1:1:1:1), and the culture is shaken at a speed of 120r / min and a culture temperature of 37±2°C for 7 days. Filter with a 0.1μm plate and frame filter to obtain a composite plant fermentation concentrate.

[0086] 4. Preparation of small molecule plant fermentation products

[0087] According to weight, 97 parts of composite plant fermentation concentrate were taken, 3 parts of 1,2-hexanediol were added, and the mixture was stirred evenly to obtain a small molecule plant fermentation product.

[0088] Example 2 Preparation of small molecule plant fermentation products

[0089] 1. Preparation of compound lactobacillus culture solution

[0090] The composite lactobacillus culture solution contains, by weight percentage, 4% of polygonatum sibiricum extract, 9% of tuckahoe sclerotium extract, 1% of avocado fruit extract, 1% of ginseng root extract and the balance of deionized water.

[0091] The above-mentioned composite lactobacillus culture solution was added into a triangular flask, mixed evenly, a cotton plug was added to the bottle mouth, and the bottle mouth was wrapped with newspaper, and sterilized at 115° C. for 20 min in a portable high-pressure sterilizer, and then taken out for use.

[0092] 2. Expansion of bacterial strains

[0093] Under sterile conditions, use an inoculation loop to pick a loop of Lactobacillus plantarum 14917 and Lactobacillus fermentans CICC25124 respectively, transfer them into different sterilized test tubes, draw a curve, and culture them in horizontal test tubes at 32°C for 7 days. After the colonies grow, transfer them again under the same operating conditions, and the culture conditions are the same as above. The strains are used in the experiment after 3 generations of activation process.

[0094] 3. Preparation of composite plant fermentation concentrate

[0095] The selected culture temperature is 37±2°C, sterile water is added to the activated slant test tube to wash out the spores, the spore suspension is aspirated with a sterile pipette, and an inoculum amount of 8% (the weight ratio of Lactobacillus plantarum 14917 and Lactobacillus fermentative mucus CICC 25124 is 1:1) is transferred into the composite lactobacillus culture solution prepared in step 1, and the culture is shaken at a speed of 120 r / min and a culture temperature of 37±2°C for 7 days. Filter with a 0.1 μm plate and frame filter to obtain a composite plant fermentation concentrate.

[0096] 4. Preparation of small molecule plant fermentation products

[0097] According to weight, 95 parts of the composite plant fermentation concentrate were taken, and 5 parts of 1,2-hexanediol were added, and the mixture was stirred evenly to obtain a small molecule plant fermentation product.

[0098] Example 3 Preparation of small molecule plant fermentation products

[0099] 1. Preparation of compound lactobacillus culture solution

[0100] The composite lactobacillus culture solution contains, by weight percentage, 11% of polygonatum sibiricum extract, 2% of tuckahoe sclerotium extract, 6% of avocado fruit extract, 6% of ginseng root extract and the balance of deionized water.

[0101] The above-mentioned composite lactobacillus culture solution was added into a triangular flask, mixed evenly, a cotton plug was added to the bottle mouth, and the bottle mouth was wrapped with newspaper, and sterilized at 115° C. for 20 min in a portable high-pressure sterilizer, and then taken out for use.

[0102] 2. Expansion of bacterial strains

[0103] Under sterile conditions, use an inoculation loop to pick a loop of Lactobacillus casei NCU011061 and Lactobacillus acidophilus NCFM respectively, transfer them into different sterilized test tubes, draw a curve, and culture them in horizontal test tubes at 32°C for 7 days. After the colonies grow, transfer them again under the same operating conditions, and the culture conditions are the same as above. The strains are used in the experiment after 3 generations of activation process.

[0104] 3. Preparation of composite plant fermentation concentrate

[0105] The selected culture temperature is 37±2°C, sterile water is added to the activated slant test tube, spores are washed off, the spore suspension is aspirated with a sterile pipette, and 2% inoculation amount (the weight ratio of human Lactobacillus casei NCU011061 and Lactobacillus acidophilus NCFM is 1:1) is transferred into the composite lactobacillus culture solution prepared in step 1, and the culture is shaken at a speed of 120r / min and a culture temperature of 37±2°C for 7 days. Filter with a 0.1μm plate and frame filter to obtain a composite plant fermentation concentrate.

[0106] 4. Preparation of small molecule plant fermentation products

[0107] According to weight, 99 parts of the composite plant fermentation concentrate were taken, 1 part of 1,2-hexanediol was added, and the mixture was stirred evenly to obtain a small molecule plant fermentation product.

[0108] Comparative Example 1 Preparation of small molecule plant fermentation products

[0109] Comparative Example 1 was prepared with reference to Example 1, and the only difference between Comparative Example 1 and Example 1 was that the composition of the composite lactobacillus culture solution was different.

[0110] The composition of the composite lactobacillus culture solution of Comparative Example 1 is as follows: 8% of polygonatum root extract, 6% of tuckahoe sclerotium extract, 3% of avocado fruit extract and the balance of deionized water.

[0111] Comparative Example 2 Preparation of small molecule plant fermentation products

[0112] Comparative Example 2 was prepared with reference to Example 1. The only difference between Comparative Example 2 and Example 1 was that the composition of the composite lactobacillus culture solution was different.

[0113] The composition of the composite lactobacillus culture solution of Comparative Example 2 is: 8% polygonatum sibiricum extract, 6% tuckahoe sclerotium extract, 3% ginseng root extract and the balance deionized water.

[0114] Comparative Example 3 Preparation of small molecule plant fermentation products

[0115] Comparative Example 3 was prepared with reference to Example 1. The only difference between Comparative Example 3 and Example 1 was that the composition of the composite lactobacillus culture solution was different.

[0116] The composition of the composite lactobacillus culture solution of Comparative Example 3: 8% polygonatum root extract, 3% ginseng root extract and the balance deionized water.

[0117] Comparative Example 4 Preparation of small molecule plant fermentation products

[0118] Comparative Example 4 was set up with reference to Example 1. The only difference between Comparative Example 5 and Example 1 is that the composition of the composite lactobacillus culture solution is different.

[0119] The composition of the composite lactobacillus culture solution of Comparative Example 4: 8% of polygonatum root extract and the balance of deionized water.

[0120] Comparative Example 5 Preparation of small molecule plant fermentation products

[0121] Comparative Example 5 is set with reference to Example 1. The difference between Comparative Example 5 and Example 1 is only that: "Step 3, preparation of composite plant fermentation concentrate" is different. The specific process is as follows:

[0122] The selected culture temperature is 37±2°C, sterile water is added to the activated slant test tube, spores are washed off, the spore suspension is aspirated with a sterile pipette, and 10% inoculum (the weight ratio of Lactobacillus plantarum 14917, Lactobacillus fermentative CICC 25124, Lactobacillus casei NCU011061 and Lactobacillus acidophilus NCFM is 1:1:1:1) is transferred into the composite lactobacillus culture solution prepared in step 1, and the culture is shaken at a speed of 120r / min and a culture temperature of 37±2°C for 7 days. Filter with a 0.1μm plate and frame filter to obtain a composite plant fermentation concentrate.

[0123] Comparative Example 6 Preparation of small molecule plant fermentation products

[0124] Comparative Example 6 is set with reference to Example 2. The difference between Comparative Example 6 and Example 2 is only that: "Step 2, bacterial culture expansion" and "Step 3, preparation of composite plant fermentation concentrate" are different. The specific process is as follows:

[0125] 2. Expansion of bacterial strains

[0126] Under sterile conditions, use an inoculation loop to pick a loop of Lactobacillus plantarum 14917 bacteria, transfer it to different sterilized test tubes, draw a curve, and culture it in a horizontal test tube at 32°C for 7 days. After the colony grows, transfer it again under the same operating conditions as above. The strain is used in the experiment after 3 generations of activation process.

[0127] 3. Preparation of composite plant fermentation concentrate

[0128] The selected culture temperature is 37±2°C, sterile water is added to the activated slant test tube, spores are washed off, the spore suspension is aspirated with a sterile pipette, and an 8% inoculum (Lactobacillus plantarum 14917) is transferred into the composite lactobacillus culture solution prepared in step 1, and the culture is shaken at a speed of 120 r / min and a culture temperature of 37±2°C for 7 days. Filter with a 0.1 μm plate and frame filter to obtain a composite plant fermentation concentrate.

[0129] Comparative Example 7 Preparation of small molecule plant fermentation products

[0130] Comparative Example 7 is set with reference to Example 2. The difference between Comparative Example 7 and Example 2 is only that: "Step 2, bacterial culture expansion" and "Step 3, preparation of composite plant fermentation concentrate" are different. The specific process is as follows:

[0131] 2. Expansion of bacterial strains

[0132] Under sterile conditions, use an inoculation loop to pick a loop of Lactobacillus plantarum NDC75017 and Lactobacillus fermentans CICC 25124 respectively, transfer them into different sterilized test tubes, draw a curve, and culture them in horizontal test tubes at 32°C for 7 days. After the colonies grow, transfer them again under the same operating conditions, and the culture conditions are the same as above. The strains are used in the experiment after 3 generations of activation process.

[0133] 3. Preparation of composite plant fermentation concentrate

[0134] The selected culture temperature is 37±2°C, sterile water is added to the activated slant test tube, spores are washed off, the spore suspension is aspirated with a sterile pipette, and an inoculum amount of 8% (the weight ratio of Lactobacillus plantarum NDC75017 and Lactobacillus fermentative mucus CICC25124 is 1:1) is transferred into the composite lactobacillus culture solution prepared in step 1, and the culture is shaken at a speed of 120 r / min and a culture temperature of 37±2°C for 7 days. Filter with a 0.1 μm plate and frame filter to obtain a composite plant fermentation concentrate.

[0135] Application Example 1-Application Example 11 Anti-aging cosmetics based on small molecule plant fermentation products

[0136] The specific formulas of Application Examples 1 to 6 are shown in Table 2, and the specific formulas of Application Examples 7 to 11 are shown in Table 3, wherein Application Example 1 does not contain small molecule plant fermentation products.

[0137] Table 2 Anti-aging cosmetic formula based on small molecule plant fermentation products in Application Examples 1 to 6

[0138]

[0139] Table 3 Application Examples 7-11 Anti-aging cosmetic formulas based on small molecule plant fermentation products

[0140]

[0141] The preparation methods of Application Examples 1 to 6 and Application Examples 9 to 11 include the following steps:

[0142] 1. Clean and disinfect the emulsifying pot;

[0143] 2. In a water pot, first add water and moisturizer into the pot, stir and heat to 80°C, keep warm for 60 minutes, stir for 5 minutes until the raw materials are completely dissolved, turn on cooling water to cool to below 38°C, add small molecule plant fermentation (Application Examples 2-6, Application Examples 9-11 all use the small molecule plant fermentation in Example 1), sodium chloride, and skin conditioner, stir (20 rpm) for 5 minutes until the raw materials are completely dissolved, measure the pH of the material, adjust the pH of the material to 6.8, and stir (20 rpm) for 5 minutes until the raw materials are completely dissolved;

[0144] 3. Add thickener, emulsifier, emollient and filler to the emulsifying pot, evacuate to -0.04MPa, stir (25 rpm), start homogenization (3000 rpm) for 8 minutes until the material is uniform, then slowly draw the water phase into the emulsifying pot, stir and disperse evenly, draw out the water phase within two to two and a half hours, stir quickly (25 rpm) for 10 minutes, homogenize evenly, then add fragrance and stir evenly;

[0145] 4. Homogenize for 2 minutes, take samples and observe the appearance. If the sample is qualified, proceed to the next step. If it is unqualified, homogenize several times and observe the appearance until the material is qualified. Then stop homogenizing.

[0146] 5. Take samples for testing. If the test is qualified, filter the material through 80 mesh;

[0147] 6. Store and age, take samples for testing, and fill, package and store qualified products.

[0148] The preparation method of Application Example 7-Application Example 8 comprises the following steps:

[0149] 1. Clean and disinfect the emulsifying pot;

[0150] 2. In a water pot, first add water and moisturizer into the pot, stir and heat to 90°C, keep warm for 30 minutes, stir for 8 minutes until the raw materials are completely dissolved, turn on cooling water to cool to below 38°C, add small molecule plant fermentation (Application Example 7 uses the small molecule plant fermentation of Example 2, and Application Example 8 uses the small molecule plant fermentation of Example 3), sodium chloride or magnesium sulfate (Application Example 7 is sodium chloride, Application Example 8 is magnesium sulfate), skin conditioner, stir (20 rpm) for 15 minutes until the raw materials are completely dissolved, measure the pH of the material, adjust the pH of the material to 5.3, and stir (20 rpm) for 10 minutes until the raw materials are completely dissolved;

[0151] 3. Add thickener, emulsifier, emollient and filler to the emulsifying pot, evacuate to -0.06MPa, stir (25 rpm), start homogenization (3000 rpm) for 15 minutes until the material is uniform, then slowly draw the water phase into the emulsifying pot, stir and disperse evenly, draw out the water phase within two to two and a half hours, stir quickly (35 rpm) for 15 minutes, homogenize evenly, then add fragrance and stir evenly;

[0152] 4. Homogenize for 2 minutes, take samples and observe the appearance. If the sample is qualified, proceed to the next step. If it is unqualified, homogenize several times and observe the appearance until the material is qualified. Then stop homogenizing.

[0153] 5. Take samples for testing. If the test is qualified, filter the material through 80 mesh;

[0154] 6. Store and age, take samples for testing, and fill, package and store qualified products.

[0155] Comparative application example 1

[0156] 1. Preparation method of oil-in-water system

[0157] Phase A: deionized water (to 100%), glycerin 10%, betaine 1%, p-hydroxyacetophenone 0.5%; Phase B: polydimethylsiloxane 5%, hydrogenated polydecene 5%, emulsifier; Phase C: thickener (SEPINOV EMT 10 (hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer, polysorbate-60, sorbitan isostearate)); Phase D: 1,2-hexanediol 0.5%, small molecule plant fermentation product of Example 1 2%.

[0158] Preparation method:

[0159] (1) Add phase B into an oil pan and stir to raise the temperature to 80°C to obtain an oil phase;

[0160] (2) Add phase A into a water pot and stir to raise the temperature to 80°C to obtain a water phase;

[0161] (3) Preheat the emulsifier and evacuate to -0.06 MPa. First draw in the water phase, then the oil phase, homogenize for 5 minutes, add phase C, homogenize for 5 minutes, stir and cool to 45°C, add phase D, stir for 20 minutes, and discharge the material after inspection.

[0162] 2. Screening test of emulsifiers and thickeners for oil-in-water system

[0163] ① Commonly used water-in-oil emulsifiers: such as cetearyl alcohol, cetearyl glucoside, glyceryl stearate or PEG-100 stearate, emulsifier addition amount 3%, thickener addition amount 1%, prepared according to the above preparation method, after adding 2% of small molecule plant fermentation product, the system changes from cream to water, indicating that the commonly used water-in-oil emulsifier system cannot achieve the expected effect.

[0164] ② Salt-tolerant water-in-oil emulsifier: A commercially available salt-tolerant phospholipid emulsifier was added at a rate of 3.5%, and a thickener was added at a rate of 1%. The product was prepared according to the above method. After adding 2% of small molecule plant fermentation products, the system did not become thinner. The samples were prepared and distributed to 15 volunteers aged between 30 and 50 years old for experience. They all agreed that the product had a thick texture and a sticky residue, which did not achieve the expected effect of product development.

[0165] Comparative Application Example 2

[0166] The only difference from Application Example 3 is the emulsifier, and the other conditions are the same, as shown in Table 4.

[0167] Table 4 Emulsifier formula for comparative application example 2

[0168]

[0169] Experimental Example 1 Anti-glycation experiment

[0170] Since the key to the formation of AGEs by glycation reaction is the condensation of the carbonyl group of reducing sugar and the free amino group of protein, we can use glucose and bovine serum albumin to generate AGEs in vitro and then add anti-glycation products to evaluate its anti-glycation effect. Most AGEs have fluorescent properties, so the fluorescence intensity can be measured by an enzyme marker to reflect the content of AGEs. By measuring the changes in the content of AGEs after the sample is administered, it is tested whether the sample has the effect of inhibiting the synthesis of AGEs. In this way, it is determined whether the sample has an anti-glycation effect. The specific experimental process is as follows:

[0171] 1) Modeling: Add the small molecule plant fermentation products (concentrations of 2.5% and 5%) prepared in Examples 1-3 and Comparative Examples 1-7, bovine serum albumin (PBS, 8.00 mg / mL) and glucose (PBS, 0.2 mol / L) mixed solution into a 96-well plate (test system: 320 μL / well); the model control group does not add the small molecule plant fermentation products, but adds an equal volume of deionized water. Incubate at 60°C for about 40 hours.

[0172] 2) Detect fluorescence intensity: After incubation, use a microplate reader to measure the fluorescence intensity at 370nm excitation wavelength and 440nm emission wavelength. Calculate the anti-glycation effect of the sample based on the fluorescence intensity. The calculation formula is as follows:

[0173] ;

[0174] Where S represents the fluorescence intensity.

[0175] The results of the test are as follows Figure 1 shown.

[0176] Experimental Example 2 Antioxidant Experiment

[0177] 1. DPPH removal experiment

[0178] Antioxidant activity can be characterized by evaluating the inhibition rate of the test sample on DPPH free radicals. DPPH analysis is a simple method for screening free radical scavengers and evaluating antioxidant activity. The ethanol solution of DPPH appears purple, with a maximum absorption wavelength of 517nm. When a free radical scavenger is added to the DPPH solution, its lone pair of electrons is paired, and the absorption disappears or weakens, causing the solution to become lighter in color, appearing yellow or light yellow, and the absorbance at 517nm becomes smaller. The degree of change is linearly related to the free radical scavenging intensity, so it can be quantitatively analyzed by spectrophotometry. This method can be expressed by the scavenging rate. The larger the scavenging rate, the stronger the scavenging ability of the substance.

[0179] DPPH determination process: A series of test sample solutions of multiple concentrations were prepared using 70% ethanol. The test sample was the small molecule plant fermentation product prepared in Example 1. 300 μL of deionized water (as a blank control), test samples of various concentrations, and VC of various concentrations (as a positive control) were added to a 96-well plate. 300 μL of DPPH working solution (solvent: anhydrous ethanol) was added to each well and mixed evenly. The absorbance was measured at 517 nm for 30 min, and the DPPH free radical scavenging rate was calculated according to the following formula.

[0180] ;

[0181] Wherein, A is the absorbance of the reaction between the sample to be tested and the DPPH working solution; A1 is the control absorbance when anhydrous ethanol replaces the DPPH working solution; A0 is the blank absorbance when 70% ethanol replaces the sample.

[0182] The results of DPPH scavenging experiment are as follows Figure 2 shown.

[0183] The half catalytic elimination efficiency (EC50) and ascorbic acid equivalent (AEAC) were calculated based on the DPPH free radical scavenging results.

[0184]

[0185] The AEAC determination results of the DPPH removal experiment are shown in Table 5:

[0186] Table 5 EC50 and AEAC results of DPPH scavenging experiment

[0187]

[0188] When the addition amount of the small molecule plant fermentation product prepared in Example 1 is 0.032%, the effect of scavenging half of the DPPH free radicals can be achieved. The antioxidant capacity of each 100g of small molecule plant fermentation product is equivalent to 190.51g VC, indicating that the small molecule plant fermentation product of the present invention has significant antioxidant capacity.

[0189] 2. ABTS removal experiment

[0190] ABTS reacts with potassium persulfate to produce green ABTS. + Free radicals. This free radical has a maximum absorption at 752nm. Therefore, by detecting the absorbance at 752nm, its concentration can be determined. + After adding free radical working solution, if the absorbance at 752nm decreases, it means that the substance has free radical scavenging activity and is an antioxidant.

[0191] ABTS determination process: Anhydrous ethanol was used to prepare a series of test sample solutions of multiple concentrations. The test sample was the small molecule plant fermentation product prepared in Example 1. 300 μL of anhydrous ethanol (as a blank control), the test sample of each concentration, and each concentration of VC (as a positive control) were mixed with 300 μL of ABTS working solution (solvent is anhydrous ethanol) and reacted for 30 minutes. The solution was transferred to a 96-well plate, the absorbance was measured at 752 nm, and the ABTS free radical scavenging rate was calculated according to the following formula.

[0192]

[0193] Wherein: A is the absorbance of the reaction between the sample to be tested and the ABTS working solution; A is the absorbance of the control when anhydrous ethanol replaces the ABTS working solution; A is the blank absorbance when anhydrous ethanol replaces the sample.

[0194] The results of ABTS clearance assay are shown in Figure 3 shown.

[0195] The half catalytic elimination efficiency (EC50) and ascorbic acid equivalent (AEAC) were calculated based on the DPPH free radical scavenging results.

[0196]

[0197] The EC50 and AEAC results of the ABTS clearance experiment are shown in Table 6:

[0198] Table 6 EC50 and AEAC results of ABTS clearance experiment

[0199]

[0200] When the addition amount of the small molecule plant fermentation product prepared in Example 1 is 0.012%, the effect of scavenging half of the ABTS free radicals can be achieved. The antioxidant capacity of each 100g of small molecule plant fermentation product is equivalent to 232.67g VC, indicating that the small molecule plant fermentation product of the present invention has significant antioxidant capacity.

[0201] Experimental Example 3 col1a1a, col1a1b and col1a2 Expression level determination

[0202] Collagen is the most abundant extracellular matrix protein in the human body, and type I collagen is the most abundant protein in the skin. The distribution of type I collagen in zebrafish is the same as that in humans and shows a high degree of conservation with humans. Their innate expression decreases significantly at 6 days after fertilization and becomes very low between 10 and 12 days after fertilization. col1a1a , col1a1b and col1a2 ) expression, compare the relative expression of type I collagen gene in zebrafish between the treatment group and the blank control group, and calculate the promotion rate of type I collagen gene expression to evaluate the anti-wrinkle and firming effects of raw materials or products. The specific experimental methods are as follows:

[0203] 1. Experimental Preparation

[0204] 1) Test organisms:

[0205] Wild-type AB strain zebrafish ( Danio Rerio ) Spawning test. The broodstock should have good reproductive ability, and the best age is 6-12 months. Cross-crossing should be used as much as possible to maintain genetic diversity. A new batch of broodstock should be replaced after 5 generations of pure strain broodstock. The broodstock should not have obvious visible signs of infection and disease, and should not have undergone drug treatment within 2 months. Before the breeding and spawning test, the broodstock should be domesticated for more than 14 days after being introduced into the laboratory.

[0206] Breeding requirements: The water temperature should be controlled at 26-28.5℃, and the indoor temperature should be controlled at 20-25℃. The breeding density should be controlled at 1-2 fish per liter of water, and the light should be fixed at 12-16 hours per day, and a good filtration system should be maintained. Feed at least 2 times a day, including at least 1 time of brine shrimp ( Artemia salina ) For larvae, the feeding interval should be more than 3 hours to avoid overfeeding that affects water quality and cleanliness.

[0207] Spawning requirements: If you use a spawning tank to collect fish eggs, put male and female fish in a ratio of 2:1 into the spawning tank 1-2 hours before turning off the lights on the day before spawning. To avoid genetic deviation, mix the fish eggs collected from at least 3 spawning tanks and select them for use. If you use a breeding fish tank to collect fish eggs, put the egg collection box into the breeding fish tank where the fish eggs are to be collected before turning off the lights on the day before spawning or before turning on the lights on the day of spawning. To prevent the eggs from being eaten by adult fish, cover the egg collection box with an inert net. Mating, spawning and fertilization are completed within about 30 minutes after turning on the lights, and the egg collection box can be removed from the fish tank at that time. After the fish eggs are removed from the egg collection box, clean the fish embryos with fish embryo culture medium. Select healthy zebrafish embryos and culture them at a density of no more than 1 fish embryo in 200μL fish embryo culture medium at 28℃±1℃ until 6 days after fertilization.

[0208] 2) Preparation of fish embryo culture medium: Weigh 2940 mg of anhydrous calcium chloride, 1233 mg of magnesium sulfate heptahydrate, 630 mg of sodium bicarbonate, and 55 mg of potassium chloride and dissolve them in 10 L of water. The pH value is 6.5-8.5. All chemicals are of analytical grade.

[0209] 3) Preparation of acetyl hexapeptide-8 solution: Use fish embryo culture medium to prepare 0.8g / L acetyl hexapeptide-8 solution, and prepare it fresh before use.

[0210] 4) Preparation of test solution: The small molecule plant fermentation product prepared in Example 1 was prepared into a test solution with a concentration of 2% using fish embryo culture medium.

[0211] 2. Experimental Grouping

[0212] The test set up a blank control group, a positive control group and a sample treatment group, and the settings of each group are as follows:

[0213] 1) Blank control group setting: 36 zebrafish were randomly selected and evenly distributed into a 24-well plate, with 3 wells, each containing 12 zebrafish and 2.5 mL of fish embryo culture medium.

[0214] 2) Positive control group setting: 36 zebrafish were randomly selected and evenly distributed into a 24-well plate, with 3 wells, each well containing 12 zebrafish and 2.5 mL of acetyl hexapeptide-8 working solution.

[0215] 3) Sample treatment group setting: 36 zebrafish were randomly selected and evenly distributed into a 24-well plate with 3 wells, each containing 12 zebrafish and 2.5 mL of the test solution.

[0216] The above groups were placed in a 28°C±1°C incubator for 24h±1h. Twelve zebrafish from each well were collected into a 1.5mL tube, the solution was removed, 0.5mL RNAlater solution was added, and the tube was frozen for storage.

[0217] 3. RNA Extraction

[0218] Remove RNAlater solution and rinse with PBS solution three times. Place in ice bath, remove PBS solution, and add 500μL TRIzol reagent. Homogenize embryos with a pellet pestle and place in ice bath for 10 minutes. Add 100μL chloroform, vortex for 1 minute, and place in ice bath for 5 minutes. Centrifuge samples for 20 minutes, transfer the upper layer solution to a 1.5mL centrifuge tube, add 250μL isopropanol, vortex for 1 minute, place in ice bath for 10 minutes, and centrifuge samples for 20 minutes. Remove all supernatant, add 500μL ethanol, vortex for 1 minute, centrifuge samples for 5 minutes, and place in ice bath for 10 minutes. Repeat this step 3 times. Remove ethanol, centrifuge samples for 5 minutes, and air-dry samples in a fume hood with the lid open for 10 minutes. Add 10μL DNase / RNase-free ultrapure distilled water and heat at 55℃ for 15 minutes.

[0219] 4. cDNA Synthesis

[0220] The RNA sample was diluted to 1000 ng / 7 μL with DNase / RNase-free ultrapure distilled water. The RNA was synthesized into cDNA using the PrimeScript RT Kit with gDNA Eraser (Takara; Cat no. RR047A) and stored frozen.

[0221] 5. Perform real-time RT-PCR

[0222] Prepare primer mix and real-time PCR master mix for each primer. Dilute cDNA sample 10-fold with DNase / RNase-free ultrapure distilled water. Add 9µL PCR master mix and 1µL diluted cDNA sample to each well of 96-well PCR plate. Seal the 96-well PCR plate with optical film, centrifuge the plate for 5 minutes, and perform real-time PCR amplification. Real-time PCR kit: SYBR Premix ExTaq (Takara; Cat no. RR420A).

[0223] 6. Calculation of results

[0224] Collect real-time PCR data. Ct is used as the amplification result. β-actin The gene amplification amount was used as the housekeeping gene, and the relative expression of each gene was calculated as the test result.

[0225] The results of the test are as follows Figure 4 As shown, the results show that the small molecule plant fermentation product of Example 1 has an effect on zebrafish at a 2% formula addition concentration. collala , collalb and col1a2The promotion rates of gene expression were 89% (p=0.0088), 108% (p=0.013) and 106% (p=0.0066), respectively. This indicates that the small molecule plant fermentation product provided by the present invention can significantly promote the expression of zebrafish type I collagen gene, promote the regeneration of type I collagen, and has anti-wrinkle and firming effects.

[0226] Experimental Example 4 Elna Expression level determination

[0227] Elastin in human skin is a protein that allows many tissues in the body to maintain their original shape after stretching and contracting, and allows the skin to return to its original shape after being squeezed. As we age, the elastin fibers in the skin will gradually break and lose, causing the skin to wrinkle and lose elasticity. Compared with human skin, zebrafish skin is highly similar to human skin in structure, except that the epidermis is differentiated epidermal cells and mucus layer instead of stratum corneum. The elastin gene is highly conserved, and the elastin in the human body is produced by Eln Gene synthesis, the corresponding gene in zebrafish is Elna , by testing whether the samples promote Elna The expression of genes can be used to determine whether the sample can promote the regeneration of elastin, help the skin resist wrinkles, and achieve skin firming. The specific experimental methods are as follows:

[0228] 1. Experimental Preparation

[0229] The experimental preparation is the same as that of Experimental Example 3.

[0230] 2. Experimental Grouping

[0231] The test set up a blank control group, a positive control group and a sample treatment group, and the settings of each group are as follows:

[0232] 1) Blank control group setting: 36 zebrafish were randomly selected and evenly distributed into a 24-well plate, with 3 wells, each containing 12 zebrafish and 2.5 mL of fish embryo culture medium.

[0233] 2) Positive control group setting: 36 zebrafish were randomly selected and evenly distributed into a 24-well plate, with 3 wells, each well containing 12 zebrafish and 2.5 mL of acetyl hexapeptide-8 working solution.

[0234] 3) Sample treatment group setting: 36 zebrafish were randomly selected and evenly distributed into a 24-well plate, with 3 wells, each well containing 12 zebrafish and 2.5 mL of the test substance (Example 1) solution.

[0235] The above groups were placed in a 28°C±1°C incubator for 24h±1h. Twelve zebrafish from each well were collected into a 1.5mL tube, the solution was removed, 0.5mL RNAlater solution was added, and the tube was frozen for storage.

[0236] 3. RNA extraction and cDNA synthesis

[0237] RNA extraction and cDNA synthesis were the same as in Experimental Example 3.

[0238] 4. Perform real-time RT-PCR

[0239] Prepare primer mix and real-time PCR master mix for each primer. Dilute cDNA sample 10-fold with DNase / RNase-free ultrapure distilled water. Add 9µL PCR master mix and 1µL diluted cDNA sample to each well of 96-well PCR plate. Seal the 96-well PCR plate with optical film, centrifuge the plate for 5 minutes, and perform real-time PCR amplification. Real-time PCR kit: SYBR Premix ExTaq (Takara; Cat no. RR420A).

[0240] 5. Result calculation

[0241] Collect real-time PCR data. Ct is used as the amplification result. β-actin The gene amplification amount was used as the housekeeping gene, and the relative expression of each gene was calculated as the test result.

[0242] The results of the test are as follows Figure 5 As shown, the sample at a 2% formula concentration was effective for zebrafish Elna The promotion rate of gene expression was 171% (p=0.00096). This shows that the small molecule plant fermentation product provided by the present invention can promote Elna Gene expression can promote the regeneration of elastin, and has anti-wrinkle and firming effects.

[0243] Experimental Example 5 Stability Test

[0244] Small molecule plant fermentation products are brown fermentation liquids. In the formula, as the concentration increases, the color of the material and the residual fermentation smell become greater. Simulating the environment and conditions of daily storage, the cosmetic creams in Application Examples 1-11 and Comparative Application Example 2 were packaged in colorless transparent sample bottles, respectively, and placed at -18°C (refrigerated conditions), 6-10°C (fresh-keeping conditions), 25°C (normal temperature conditions), 45°C (high temperature conditions), and -18°C and 45°C cycles (one day of refrigeration and one day of high temperature cycles) for 2 months, and the appearance was continuously observed; the degree of change is expressed by 0-3, 0 is no obvious change in color, texture, and smell, 1 is an acceptable slight change (such as slight discoloration, slightly thickened texture, lighter fragrance / slightly changed taste), 2 is an unacceptable change (such as obvious discoloration, sticky and uneven texture, serious odor change / obvious raw material smell), 3 is a serious change (such as serious reduction in volume, yellow-brown color, and pungent raw material smell in the smell). The observation results are shown in Table 7.

[0245] Table 7 Stability of each application example

[0246]

[0247] From the experimental results, it can be seen that the increase in temperature will accelerate the changes in the color and smell of the material, and as the concentration ratio of small molecule plant fermentation in the formula increases, the impact on the color and smell changes of the formula material will be greater. When the addition amount of small molecule plant fermentation is 0.5%-5% (Application Example 2-Application Example 5), the color and smell changes of the material are in an acceptable state. When the addition amount of small molecule plant fermentation is 5%, the product material has a darker color and a more obvious change in smell under long-term high temperature, but it is still within the acceptable range. When the addition amount of small molecule plant fermentation is 6% (Application Example 6), the product material has a serious darkening of color, a significant change in smell, and a pungent smell of raw materials under long-term high temperature, which is an unacceptable change. Therefore, the formula recommends that the addition amount of small molecule plant fermentation is 0.5%-5%.

[0248] The emulsifier composition of comparative application example 2 has changed and the stability has decreased, indicating that the combination of Winsier Plus, cetyl PEG / PPG-10 / 1 polydimethylsiloxane and lauryl PEG-8 polydimethylsiloxane can improve the stability of the product.

[0249] Experimental Example 6 Evaluation of long-term anti-aging effect

[0250] 1) Test samples: samples of Application Examples 1 to 8.

[0251] 2) Test purpose: To evaluate the anti-aging efficacy of the test sample by having adult female volunteers use the test sample continuously under normal conditions.

[0252] 3) Volunteers: Age: 35-60 years old; Gender: female; Number: 160 people.

[0253] 4) Volunteer facial skin condition: dull facial complexion, expression lines, and fine lines.

[0254] 5) Product usage frequency: Apply evenly after cleansing, twice a day, once in the morning and once in the evening, for four weeks.

[0255] 6) Test item: long-term anti-aging effect.

[0256] 7) Evaluation method: A questionnaire survey was conducted to understand the subjects' evaluation of the improvement of wrinkles after using the product for 4 weeks. The evaluation index was mainly: wrinkle lightening effect. The results were divided into: not obvious effect, general effect, obvious effect, and very obvious effect. The specific test results are shown in Table 8 below.

[0257] Table 8 Long-lasting anti-aging effect

[0258]

[0259] Experimental conclusion: The statistical results of the wrinkle-lightening effect of the product after 28 days of trial use by volunteers in each experimental group showed that adding small molecule plant fermentation products can make the product have a wrinkle-lightening effect, and as the amount added increases, the wrinkle-lightening effect becomes more obvious.

[0260] Experimental Example 7 Consumer Skin Feeling Experience Test

[0261] (1) Test method:

[0262] 20 female volunteers with an average age of 30±5 years old were selected and provided with the application example cosmetics of the present invention for trial use.

[0263] (2) Evaluation method:

[0264] ①Apply an appropriate amount on the face and gently massage with your fingers until it is completely absorbed;

[0265] ② After 15 minutes, rate the product based on moisturizing, comfort, gentleness, and purchase intention and fill out the form.

[0266] (3) Scoring criteria:

[0267] Each item is scored from 1 to 5. The rating scale is: "1" means very dissatisfied; "2" means dissatisfied; "3" means average; "4" means relatively satisfied; and "5" means very satisfied.

[0268] (4) Evaluation results:

[0269] The evaluation results are shown in Table 9.

[0270] Table 9 Consumer skin experience results

[0271]

[0272] From the feedback on consumers' skin feel experience in Table 9, it can be seen that the overall skin feel of this formula is light, non-sticky, easy to absorb and non-irritating. The oil film formed on the skin after application has a certain moisturizing feeling. As the concentration of small molecule plant fermentation products increases, the moisturizing feeling is enhanced. Consumers use odor and oil film residue to score the comfort. As the concentration of small molecule plant fermentation products increases, the comfort of the experiencers decreases. This is based on the preference of women in the 30±5 age group for a skin feel experience with low residual feeling.

[0273] The emulsifier composition of comparative application example 2 changed, and the consumer skin feel experience score decreased, indicating that the combination of Winsier Plus, cetyl PEG / PPG-10 / 1 polydimethylsiloxane and lauryl PEG-8 polydimethylsiloxane can improve the skin feel experience of the product.

[0274] The above detailed description is a specific description of one feasible embodiment of the present invention. The embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or modification that does not deviate from the present invention should be included in the scope of the technical solution of the present invention.

Claims

1. An anti-aging cosmetic containing small molecule plant fermentation products, characterized in that: In terms of mass percentage, including Oil phase: Thickener 0.5%-2%; Emulsifier 3%-5%; Emollient 5%-35%; Filler 0.1%-3%; Aqueous phase: Small molecule plant fermentation 0.5%-5%; Moisturizer 10%-20%; Sodium chloride or magnesium sulfate 0.5%-1.5%; Skin conditioner 0.1%-2%; Fragrance 0.01%-0.5%; Water balance; The pH of the aqueous phase is controlled within the range of 5.0-7.0; The small molecule plant fermentation product comprises, by weight: 95-99 parts of composite plant fermentation concentrate and 1-5 parts of protective agent; The composite plant fermentation concentrate uses polygonatum root extract, tuckahoe sclerotium extract, avocado fruit extract, and ginseng root extract as composite plant raw materials, and is fermented with composite lactobacillus to obtain secondary metabolites; The composite plant fermentation concentrate is prepared from the following raw materials in weight percentage: 4%-11% polygonatum root extract, 2%-9% tuckahoe sclerotium extract, 1%-6% avocado fruit extract, 1%-6% ginseng root extract, 2%-8% composite lactobacillus and the balance water; The composite lactobacillus is composed of at least two of Lactobacillus plantarum, Lactobacillus mucus fermentation, Lactobacillus casei and Lactobacillus acidophilus; The anti-aging cosmetic is a water-in-oil system; The emulsifier includes Winsier Plus, cetyl PEG / PPG-10 / 1 dimethicone, and lauryl PEG-8 dimethicone, and the Winsier Plus includes PEG-10 dimethicone, phenyl dimethicone, and disteardimonium hectorite; The emollients include silicone oil emollients and grease emollients.

2. The anti-aging cosmetic according to claim 1, characterized in that: The pH adjuster used to adjust the pH of the aqueous phase includes at least one of sodium hydroxide, citric acid, aminomethyl propanol and triethanolamine.

3. The anti-aging cosmetic according to claim 1, characterized in that: The fragrance includes at least one of essential oil and essence.

4. The anti-aging cosmetic according to claim 1, characterized in that: The skin conditioning agent includes one or more of glycerophosphoinositol choline salt, palmitoyl tripeptide-5, hydrolyzed collagen, ferulic acid, ergothioneine, acetyl tetrapeptide-2, hydroxypropyl tetrahydropyrantriol, tetrahydromethylpyrimidine carboxylic acid, carbohydrate isomers, p-hydroxyacetophenone, ceramide NP, sodium hyaluronate, allantoin, dipotassium glycyrrhizinate, acetyl hexapeptide-8, acetylated sodium hyaluronate and soluble collagen.

5. The anti-aging cosmetic according to claim 1, characterized in that: The silicone oil emollient includes at least one of polydimethylsiloxane and cyclopentasiloxane; the oil emollient includes at least one of ethylhexyl palmitate, isohexadecane, caprylic / capric triglyceride, tri(ethylhexanoin), hydrogenated polydecene and neopentyl glycol dicaprylate / dicaprate.

6. The anti-aging cosmetic according to claim 1, characterized in that: The protective agent includes: one or more of 1,2-hexanediol, 1,2-pentanediol, caprylyl glycol, ethylhexylglycerol; and / or The preparation method of the small molecule plant fermentation product comprises: mixing polygonatum root extract, tuckahoe sclerotium extract, avocado fruit extract, ginseng root extract and water, adding the cultured composite lactobacillus, filtering to obtain a filtrate after fermentation, adding a protective agent, and stirring evenly to obtain the product.

7. The anti-aging cosmetic according to claim 1, characterized in that: The humectant includes one or more of glycerin, butylene glycol, 1,2-hexanediol, 1,2-propylene glycol, 1,3-propylene glycol, 1,2-pentanediol, ethylhexylglycerin and HTsoft_5688, and the HTsoft_5688 includes dipropylene glycol, p-hydroxyacetophenone, 1,2-hexanediol, caprylyl glycol and isopentyl glycol.

8. The anti-aging cosmetic according to claim 1, characterized in that: The thickener is quaternary ammonium salt-18 bentonite.

9. The anti-aging cosmetic according to claim 1, characterized in that: The filler is silica.

10. The anti-aging cosmetic according to claim 1, characterized in that: The mass ratio of Winsier Plus, cetyl PEG / PPG-10 / 1 polydimethylsiloxane and lauryl PEG-8 polydimethylsiloxane is 1-6:1:

1.

11. A method for preparing the anti-aging cosmetic according to any one of claims 1 to 10, characterized in that: The steps include: (1) Add water and moisturizer into a water pot, heat up, stir until completely dissolved, cool down, add small molecule plant fermentation, sodium chloride or magnesium sulfate, and skin conditioner, stir until completely dissolved, adjust the pH to within the range of 5.0-7.0, stir until completely dissolved, and obtain an aqueous phase; (2) Add thickener, emulsifier, emollient and filler into the emulsifying pot, stir and homogenize under vacuum conditions, add water phase after the material is uniform, stir and homogenize under vacuum conditions, then add fragrance and stir evenly.

12. The preparation method according to claim 11, characterized in that: The heating temperature in step (1) is 80-90°C, and the temperature is kept for 30-60 minutes after heating; the cooling temperature is below 38°C; and / or the vacuum condition in step (2) is -0.04MPa to -0.06MPa.

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