Composition of gamma-polyglutamic acid and small molecule peptide for beauty makeup products

Abstract

The invention belongs to the field of biological medicine, and particularly discloses a gamma-polyglutamic acid and small molecule peptide composition of a beauty makeup product and a preparation process of the gamma-polyglutamic acid and small molecule peptide composition. The composition comprises the following components in parts by weight: 5-50 parts of gamma-polyglutamic acid, 1-25 parts of plant extracted small molecule peptide with a unique amino acid sequence, a humectant, a thickening agent, a preservative, a pH regulator and deionized water. The amino acid sequence of the small molecule peptide is not reported, and the small molecule peptide can activate a cell-related signal channel. The preparation process comprises the steps of plant sample roughing, small molecule peptide extraction, purification and sequencing, gamma-polyglutamic acid pretreatment, composition mixing and quality detection. Through innovative component combination and process, synergistic interaction is realized, and a beauty makeup product with remarkable effect and stable quality can be prepared.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine and beauty products, and particularly to a composition of γ-polyglutamic acid and small molecule peptides for beauty products and a preparation process thereof. Background Art

[0002] With the improvement of people's living standards, the demand for beauty products is becoming increasingly diverse. Consumers not only focus on the basic beauty effects of products, such as moisturizing, whitening, and anti-wrinkle, but also put forward higher requirements for the safety, naturalness, and lasting efficacy of products. As a biodegradable natural polymer, γ-polyglutamic acid has excellent moisturizing properties. Its unique molecular structure can bind a large number of water molecules, forming a moisturizing barrier on the skin surface to prevent water loss and keep the skin moisturized. At the same time, small molecule peptides are favored in the beauty field because of their small molecular weight and easy absorption by the skin, and can effectively participate in the physiological activities of skin cells, promote collagen synthesis, regulate cell metabolism, etc.

[0003] At present, in the beauty products on the market, although there are already some products containing γ-polyglutamic acid or small molecule peptides, there are many deficiencies. On the one hand, most of the existing small molecule peptides are common amino acid sequence combinations. After long-term research and extensive application, their efficacy improvement space is limited, and it is difficult to meet the needs of consumers for more efficient skin care ingredients. On the other hand, there are few products that rationally combine γ-polyglutamic acid and small molecule peptides and optimize the preparation process to achieve synergistic effects. Most products are simply mixed, failing to fully exert the potential of both, and may have problems such as reduced ingredient activity and poor stability during the preparation process, affecting the product quality and use effect. Therefore, it is of great practical significance to develop a plant-extracted small molecule peptide with a new amino acid sequence, scientifically combine it with γ-polyglutamic acid, and optimize the preparation process to obtain a beauty product composition with better efficacy and stability. Summary of the Invention

[0004] The present invention provides a composition of γ-polyglutamic acid and related small molecule peptides for beauty products, including the following components: 5-50 parts of γ-polyglutamic acid, 1-25 parts of plant-extracted small molecule peptides, 20-150 parts of a moisturizer, 1-15 parts of a thickener, 1-5 parts of a preservative, 0.1-2.5 parts of a pH regulator, and the balance being deionized water.

[0005] Further, the plant-extracted small molecule peptide has at least one of the following amino acid sequences:

[0006] Amino acid sequence one: Ala-Gly-Ser-Tyr-Pro-His-Cys-Val-Gln-Thr-Leu-Asn;

[0007] Amino acid sequence two: Leu-Ile-Phe-Lys-Asp-Glu-Trp-Met-Pro-Gly-Thr-Ser;

[0008] Amino acid sequence three: Cys-Gln-Tyr-His-Asn-Val-Leu-Ala-Pro-Ser-Thr-Gly;

[0009] Amino acid sequence four: Gly-Pro-Hyp-Lys-Arg-Ser-Asp-Glu-Thr-Ala-Val-Ile.

[0010] Furthermore, the γ-polyglutamic acid is a homopolymer with a molecular weight between 500 kDa and 1500 kDa, and is prepared by the microbial fermentation method.

[0011] Furthermore, the humectant is one or a combination of glycerol, butanediol, and sodium hyaluronate.

[0012] Furthermore, the thickener is one or more of xanthan gum, carbomer, and hydroxyethyl cellulose.

[0013] Furthermore, the preservative is one or more of phenoxyethanol, methyl paraben, and potassium sorbate.

[0014] Furthermore, the pH regulator is one or more of citric acid, sodium citrate, and triethanolamine, and the pH value of the composition is 5.0 - 7.0.

[0015] On the other hand, the preparation process of the composition of γ-polyglutamic acid and related small molecule peptides for beauty products is characterized by including the following steps:

[0016] Coarse selection of plant-extracted small molecule peptides: Collect plant samples with potential skin care effects in different ecological environments, establish a plant sample library, and select plants that may be rich in small molecule peptides with special biological activities based on traditional medicinal records, local folk use experience, and preliminary cell activity tests;

[0017] Extraction and purification of plant-extracted small molecule peptides: Select specific plant raw materials determined by coarse selection, wash, dry, and then crush them into fine powder. Use supercritical carbon dioxide extraction technology for preliminary extraction, then purify through a macroporous adsorption resin column, and further remove impurities and macromolecular substances through ultrafiltration technology to obtain a high-purity plant-extracted small molecule peptide solution, which is freeze-dried and reserved;

[0018] Sequencing of small molecule peptides extracted from plants: Dissolve the freeze-dried small molecule peptide sample in an appropriate amount of buffer, and use liquid chromatography-mass spectrometry (LC-MS / MS) technology for sequencing analysis. Combine bioinformatics software with a self-built database for comparison to determine the amino acid sequence;

[0019] Pretreatment of γ-polyglutamic acid: Dissolve the γ-polyglutamic acid powder prepared by microbial fermentation in deionized water to prepare a solution with a mass concentration of 12%-18%. Remove the impurity ions therein through an ion exchange resin column, and then perform vacuum concentration until the mass concentration of the solution reaches 32%-38% for standby;

[0020] Preparation of the composition: Add 80% of the total amount of deionized water to the reaction kettle equipped with a stirring device, start stirring, slowly add the thickener, stir evenly to make it fully swell, and then add the humectant, the pretreated γ-polyglutamic acid solution, and the small molecule peptide extracted from plants in sequence. Continue stirring to fully mix the components. Then add the preservative, adjust the pH value of the composition to 5.0-7.0 with a pH regulator after stirring, and finally add the remaining 20% of deionized water to make up the volume and stir evenly;

[0021] Quality inspection: Conduct inspection items such as appearance, odor, pH value, viscosity, microbial indicators, and active ingredient content on the prepared composition. Package and store the qualified products.

[0022] Furthermore, the extraction pressure of the supercritical carbon dioxide extraction technology is 25-35 MPa, the extraction temperature is 40-50 °C, and ethanol (concentration 55%-65%) is used as an entrainer to assist extraction for 2.5-3.5 hours.

[0023] Furthermore, the temperature of the vacuum concentration is controlled at 45-55 °C.

[0024] Beneficial effects: The present invention extracts small molecule peptides with unreported amino acid sequences from specific plants for the first time. These unique sequences endow the small molecule peptides with new biological activities and can interact with skin cells more effectively. Different amino acid sequences have different skin care effects, such as promoting collagen synthesis, antioxidant, maintaining skin acid-base balance, promoting cell renewal and repair, increasing skin elasticity and firmness, etc. Compared with traditional small molecule peptides, it has more significant effects in anti-wrinkle, firming the skin, antioxidant, improving skin color and texture, etc.

[0025] The γ-polyglutamic acid is scientifically combined with new plant-derived small molecule peptides. The moisturizing property of γ-polyglutamic acid and the various cell-regulating functions of small molecule peptides act synergistically. While forming a moisturizing barrier on the skin surface, it promotes skin cell metabolism, resists oxidative stress, improves skin structure and function from multiple aspects, and enhances the overall skin condition. At the same time, ingredients such as moisturizers, thickeners, preservatives, and pH regulators cooperate with each other to optimize the product performance. For example, moisturizers enhance the moisturizing effect, thickeners improve the spreadability of the product, preservatives ensure the safety of the product, and pH regulators maintain an appropriate pH value to improve the stability and efficacy of the product.

[0026] In the preparation process, starting from the rough selection of plant samples, technologies such as supercritical carbon dioxide extraction, macroporous adsorption resin purification, ultrafiltration, and advanced LC-MS / MS sequencing are used to extract and determine plant-derived small molecule peptides, retaining their active ingredients and unique structures to the greatest extent. The γ-polyglutamic acid is pretreated by ion exchange resin impurity removal and vacuum concentration to improve its purity and quality. During the preparation of the composition, parameters such as the addition sequence, stirring speed, and time of each ingredient are strictly controlled to ensure the full mixing of each ingredient, improve the quality stability and uniformity of the product, and thus provide consumers with beauty products with excellent efficacy and reliable quality. Brief Description of the Drawings

[0027] Figure 1 Overall method principle flow chart. Detailed Description of the Invention

[0028] Example 1

[0029] Composition formula

[0030] By weight, 10 parts of γ-polyglutamic acid, 5 parts of plant-derived small molecule peptide (amino acid sequence one), 30 parts of glycerol, 2 parts of xanthan gum, 1 part of phenoxyethanol, 0.2 part of citric acid, and the balance is deionized water.

[0031] Preparation process

[0032] 1. Rough selection of plant-derived small molecule peptides: Collect various algae samples from deep-sea areas. After screening, Sargassum is selected, which has an obvious promoting effect on the proliferation of skin fibroblasts and enters the extraction process.

[0033] 2. Extraction and purification of plant-derived small molecule peptides: Select Sargassum, wash, dry, and crush it. Using supercritical carbon dioxide extraction technology, under the conditions of an extraction pressure of 30 MPa and an extraction temperature of 45 °C, 60% ethanol is used as an entrainer for extraction for 3 hours to obtain a crude extract. The crude extract is passed through an AB-8 type macroporous adsorption resin column and eluted successively with water, 30% ethanol, and 50% ethanol. The 50% ethanol eluate is collected, ultrafiltered through a 5000 Da ultrafiltration membrane, and freeze-dried to obtain small molecule peptides.

[0034] 3. Sequencing of small molecule peptides extracted from plants: LC-MS / MS technology combined with Mascot software analysis was used to determine whether the amino acid sequence of the small molecule peptide met the requirements of the invention.

[0035] 4. Pretreatment of γ-polyglutamic acid: prepare γ-polyglutamic acid powder into a 15% solution, pass it through a strong acid cation exchange resin and a strong basic anion exchange resin column to remove impurities, and concentrate it under reduced pressure at 45°C to a concentration of 35% for use.

[0036] 5. Preparation of the composition: Add 80% deionized water to the reactor, stir at 150r / min, add xanthan gum to swell for 1.5 hours, add glycerol, γ-polyglutamic acid solution, and small molecule peptide in sequence, stir for 45 minutes, add phenoxyethanol and stir for 20 minutes, adjust the pH to 6.0 with citric acid, add the remaining deionized water to make up to volume, and stir evenly.

[0037] 6. Quality inspection: After testing, the product has a uniform and delicate appearance, no odor, a pH value of 6.0, appropriate viscosity, qualified microbial indicators, and the content of γ-polyglutamic acid and small molecule peptides meets the standards.

[0038] Comparative efficiency analysis

[0039] 1. Comparison with the use of γ-polyglutamic acid alone: ​​When γ-polyglutamic acid is used alone, it mainly forms a moisturizing barrier on the skin surface and increases the water content of the skin's stratum corneum. However, in a 4-week volunteer trial experiment, the control group that used the γ-polyglutamic acid product alone had a 20% increase in skin moisture content and a 10% increase in elasticity. After adding a small molecule peptide with a specific amino acid sequence 1 in Example 1, the skin moisture content of the experimental group increased by 35% and skin elasticity increased by 28%. This is because small molecule peptides can activate the intracellular signaling pathway that promotes collagen synthesis, promote fibroblasts to synthesize more collagen fibers, and enhance skin elasticity. At the same time, its role in promoting cell metabolism, in turn, enhances the skin's own moisturizing ability, further consolidates the moisturizing effect of γ-polyglutamic acid, and improves skin condition in all directions, with an effect far exceeding the use of γ-polyglutamic acid alone.

[0040] 2. Comparison with the use of glycerol alone: When glycerol is used alone, it mainly relies on the three hydroxyl groups in the molecule to form hydrogen bonds with water molecules, absorbing moisture from the surrounding environment and retaining it on the skin surface. In the skin moisture loss experiment, after the sample containing only glycerol was placed in a dry environment for 8 hours, the moisture loss rate reached 30%. When glycerol and small molecule peptides act synergistically in Example 1, the small molecule peptides regulate the expression of aquaporins in skin cells, promote the balanced transport of water inside and outside the cells, enhance the uptake and utilization efficiency of the water adsorbed by glycerol by the cells, enable the moisturizing effect of glycerol on the skin surface to be better transmitted to the interior of the cells. At the same time, the new cells generated by the small molecule peptides promoting cell metabolism have stronger water retention ability, echoing the moisturizing network constructed by glycerol. The moisture loss rate of the sample containing the composition of small molecule peptides and glycerol is only 18%, significantly strengthening the skin's moisturizing barrier and improving the durability and effect of moisturization.

[0041] 3. Comparison with the use of xanthan gum alone: When xanthan gum is used alone, it can regulate the rheological properties of the product, giving the product an appropriate consistency and certain spreadability. However, in the consumer use experience survey, for the product containing only xanthan gum without small molecule peptides, although the spreadability is good, the feedback on the improvement of skin efficacy is only 50%. In Example 1, when xanthan gum and small molecule peptides are combined, the stable system formed by xanthan gum helps the small molecule peptides to be evenly dispersed in the product, preventing their aggregation and precipitation, and ensuring the stability of small molecule peptides during storage and use. At the same time, the improved spreadability of the product by xanthan gum enables the small molecule peptides to cover the skin surface more evenly, increasing the contact area with skin cells, thereby improving the absorption efficiency and efficacy of small molecule peptides. 90% of the volunteers reported that the product containing the composition of small molecule peptides and xanthan gum had smooth spreading and more obvious skin improvement effect after use.

[0042] 4. Comparison with the use of phenoxyethanol alone: When phenoxyethanol is used alone, it can effectively inhibit the growth of common bacteria, molds and yeasts, preventing the product from being contaminated by microorganisms. In the product microbial stability test, for the product containing only phenoxyethanol, the retention rate of small molecule peptide activity is only 60%. When phenoxyethanol and small molecule peptides act synergistically in Example 1, the small molecule peptides themselves have certain antioxidant ability, which can reduce the deterioration of product components caused by oxidation, reduce the risk of microbial growth, and assist phenoxyethanol to better exert its antibacterial effect. At the same time, the sterile environment created by phenoxyethanol is conducive to maintaining the biological activity of small molecule peptides, preventing them from being degraded by microorganisms or inactivated due to the influence of microbial metabolites. After the product containing the composition of small molecule peptides and phenoxyethanol is stored under accelerated aging conditions for 6 months, the microbial indicators still meet the standards, and the retention rate of small molecule peptide activity reaches 85%.

[0043] 5. Comparison with the pH adjustment by citric acid alone: Citric acid alone can adjust the pH value of the product, but the biological activity of small molecule peptides is relatively sensitive to the environmental pH value. In in vitro cell experiments, when the pH value is adjusted only by citric acid and deviates from the range of 5.5 - 6.5, the expression level of genes related to collagen synthesis in the cell culture medium added with small molecule peptides increases limitedly. In Example 1, citric acid is used to adjust the pH to 6.0, which is within the pH range suitable for the activity of small molecule peptides, ensuring that the small molecule peptides are always in the best active state and fully exerting their efficacy in promoting cell metabolism and collagen synthesis. Compared with adjusting the pH with citric acid alone, the efficacy of small molecule peptides in beauty products is greatly improved.

[0044] Example 2

[0045] Composition formula

[0046] By weight, 20 parts of γ-polyglutamic acid, 10 parts of plant-extracted small molecule peptide (amino acid sequence two), 25 parts of butanediol, 3 parts of carbomer, 1.5 parts of methyl p-hydroxybenzoate, 0.5 part of sodium citrate, and the balance is deionized water.

[0047] Preparation process

[0048] 1. Preliminary selection of plant-extracted small molecule peptides: Collect various rare herbaceous plants in alpine regions. After preliminary screening based on traditional medicinal records and cell activity, Rhodiola cretinii is selected for subsequent extraction.

[0049] 2. Extraction and purification of plant-extracted small molecule peptides: Select Rhodiola cretinii. After treatment, supercritical carbon dioxide extraction is used, with an extraction pressure of 25 MPa and a temperature of 40 °C, and 55% ethanol as the entrainer for 2.5 hours. The crude extract is purified by a macroporous adsorption resin column, ultrafiltered, and then freeze-dried to obtain small molecule peptides.

[0050] 3. Sequencing of plant-extracted small molecule peptides: Use LC-MS / MS and PEAKS software for sequencing analysis to confirm the small molecule peptide sequence.

[0051] 4. Pretreatment of γ-polyglutamic acid: Prepare a 18% solution of γ-polyglutamic acid, remove impurities with ion exchange resin, and concentrate it to 38% under reduced pressure at 50 °C for standby.

[0052] 5. Preparation of the composition: Add 80% deionized water to the reaction kettle, add carbomer and swell it for 1.2 hours under stirring at 180 r / min, then add butanediol, γ-polyglutamic acid solution, and small molecule peptides in sequence, stir for 50 minutes, add methyl p-hydroxybenzoate and stir for 25 minutes, adjust the pH to 5.5 with sodium citrate, and make up the volume and stir evenly.

[0053] 6. Quality inspection: product appearance and smell are normal, pH value is 5.5, viscosity meets the requirements,

[0054] No pathogenic bacteria were detected in the microorganisms and the content of active ingredients met the standards.

[0055] Comparative efficiency analysis

[0056] 1. Comparison with γ-polyglutamic acid alone: ​​The control group using γ-polyglutamic acid alone had limited effect in improving skin moisture and elasticity. In Example 2, due to the addition of a small molecule peptide with amino acid sequence 2, tryptophan (Trp) and phenylalanine (Phe) in its sequence have antioxidant properties, and lysine (Lys) and arginine (Arg) help maintain the acid-base balance of the skin, synergizing with γ-polyglutamic acid. γ-Polyglutamic acid provides surface moisturizing, and small molecule peptides improve the skin at the cellular level. In the trial of volunteers, the product of Example 2 increased the moisture content of the skin by a greater extent than the group using γ-polyglutamic acid alone, and performed better in improving the skin's resistance to free radical damage and maintaining acid-base balance, and the overall effect was significantly improved.

[0057] 2. Comparison with butanediol used alone: ​​When butanediol acts alone, it assists in moisturizing through the hydroxyl groups in the molecule. In Example 2, small molecule peptides work together with butanediol. Small molecule peptides regulate the expression of cellular water channel proteins, making the cells more efficient in absorbing the skin surface moisture maintained by butanediol, and enhancing the water retention capacity of newly generated cells, and together with butanediol, a more stable moisturizing system is constructed. In the skin water loss test, the water loss rate of the sample of the product in Example 2 was significantly lower than that of the sample using only butanediol, and the moisturizing effect was greatly enhanced.

[0058] 3. Comparison with carbomer alone: ​​Carbomer alone can improve the spreadability of the product. In Example 2, carbomer and small molecule peptides work together, and the stable system allows the small molecule peptides to be evenly dispersed. The good spreadability increases the contact between the small molecule peptides and skin cells. Consumers reported that the product of Example 2 is not only smooth to apply, but also significantly better than products containing only carbomer in terms of anti-oxidation and maintaining skin acid-base balance, indicating that the small molecule peptides play a better role with the help of the characteristics of carbomer.

[0059] 4. Compared with the use of methyl parahydroxybenzoate alone: ​​When methyl parahydroxybenzoate is used alone for antibacterial purposes, the activity of small molecule peptides is easily affected. In Example 2, the antioxidant properties of small molecule peptides reduce the risk of microbial growth and assist methyl parahydroxybenzoate in antibacterial activities. At the same time, the antibacterial environment ensures the activity of small molecule peptides. In the product microbial stability test, the small molecule peptide activity retention rate of the product in Example 2 is higher than that of the product containing only methyl parahydroxybenzoate, and both the product quality and the activity of small molecule peptides are better guaranteed.

[0060] 5. Comparison with adjusting pH using sodium citrate alone: When adjusting pH using sodium citrate alone, if the pH is not appropriate, the activity of small molecule peptides is limited. In Example 2, the pH was adjusted to 5.5, which is within the optimal range for the activity of small molecule peptides. The effects such as antioxidant activity and maintaining the skin's acid-base balance of small molecule peptides were fully exerted. Compared with adjusting pH using sodium citrate alone, the product has a better effect in improving the skin condition.

[0061] Example 3

[0062] Composition formula

[0063] By weight, 15 parts of γ-polyglutamic acid, 8 parts of small molecule peptides extracted from plants (amino acid sequence three), 20 parts of sodium hyaluronate, 2 parts of hydroxyethyl cellulose, 1.2 parts of potassium sorbate, 0.3 parts of triethanolamine, and the balance is deionized water.

[0064] Preparation process

[0065] 1. Preliminary selection of small molecule peptides extracted from plants: Algae samples were collected from deep sea areas, and Laminaria japonica was selected for extracting small molecule peptides.

[0066] 2. Extraction and purification of small molecule peptides extracted from plants: Select Laminaria japonica, wash, dry, and crush it, then use supercritical carbon dioxide extraction with an extraction pressure of 32 MPa and a temperature of 42 °C, and use 62%

[0067] ethanol as the entrainer for extraction for 3 hours. Subsequently, pass through an AB-8 type macroporous adsorption resin column, elute with ethanol of different concentrations, collect the eluate containing the target small molecule peptides, then ultrafilter through a 5000 Da ultrafiltration membrane, and freeze-dry to obtain small molecule peptides.

[0068] 3. Sequencing of small molecule peptides extracted from plants: Using LC-MS / MS technology combined with a self-built database, and using PEAKS software to determine that the amino acid sequence of the small molecule peptide is amino acid sequence three in the invention.

[0069] 4. Pretreatment of γ-polyglutamic acid: Prepare a 16% solution of γ-polyglutamic acid powder, and successively pass through strong acidic cation exchange resin and strong basic anion exchange resin columns to remove impurity ions, and concentrate under reduced pressure to 36% concentration at 48 °C for standby.

[0070] 5. Preparation of the composition: Add 80% deionized water to the reaction kettle, stir at a speed of 160 r / min, add hydroxyethyl cellulose to swell for 1.3 hours, successively add sodium hyaluronate, γ-polyglutamic acid solution, and small molecule peptides, stir for 48 minutes, add potassium sorbate and stir for 22 minutes, adjust the pH to 5.8 with triethanolamine, add the remaining 20% deionized water to make up the volume, and stir evenly.

[0071] 6. Quality inspection: The product appearance is uniformly translucent, odorless, with a pH value of 5.8, the viscosity meets the design requirements, the microbial indicators are qualified, and the contents of γ-polyglutamic acid and small molecule peptides reach the standard.

[0072] Analysis of comparative synergistic effect

[0073] 1. Comparison with the sole use of γ-polyglutamic acid: When γ-polyglutamic acid is used alone, it mainly focuses on skin moisturization. In Example 3, the small molecule peptide of amino acid sequence three is added, and the sulfhydryl group of cysteine (Cys) participates in the antioxidant defense and regulates cell gene expression to promote renewal and repair. After synergizing with γ-polyglutamic acid, not only the moisturizing effect is enhanced due to the small molecule peptide promoting cell metabolism, but also better effects are achieved in improving skin color and texture. In the feedback from volunteers, the product of Example 3 makes the skin more moisturized, with more uniform color and finer texture, which is better than the effect of using γ-polyglutamic acid alone.

[0074] 2. Comparison with the sole use of sodium hyaluronate: When sodium hyaluronate is used alone, it can bind a large amount of water to form a moisturizing gel on the skin surface. In Example 3, the small molecule peptide synergizes with sodium hyaluronate. The small molecule peptide regulates the absorption and utilization of water by cells, enabling the moisturizing effect of sodium hyaluronate to better penetrate to the cell level. At the same time, the new cells generated by promoting cell metabolism and sodium hyaluronate jointly strengthen the moisturizing network. In the test of skin moisture content, the product of Example 3 has stronger skin moisture retention ability and more lasting moisturizing effect.

[0075] 3. Comparison with the sole use of hydroxyethyl cellulose: When hydroxyethyl cellulose is used alone, it can regulate the rheological properties of the product. In Example 3, hydroxyethyl cellulose synergizes with the small molecule peptide to ensure the uniform distribution of the small molecule peptide. Its improved spreadability helps the small molecule peptide to contact skin cells more comprehensively. User experience shows that when using the product of Example 3, the application process is comfortable, and the effects in antioxidant and improving skin color and texture are significantly better than those of the product containing only hydroxyethyl cellulose, reflecting the promotion of the small molecule peptide's efficacy by their synergy.

[0076] 4. Comparison with the sole use of potassium sorbate: When potassium sorbate inhibits bacteria alone, the activity of the small molecule peptide may be damaged. In Example 3, the antioxidant ability of the small molecule peptide assists potassium sorbate in inhibiting the growth of microorganisms, and the sterile environment created by potassium sorbate maintains the activity of the small molecule peptide. In the test of product microbial stability, the small molecule peptide activity retention rate of the product of Example 3 is higher, and the quality and small molecule peptide activity within the product shelf life are guaranteed.

[0077] 5. Comparison with adjusting pH with triethanolamine alone: If the pH is not adjusted to the appropriate range when using triethanolamine alone to adjust pH, the activity of small molecule peptides will be affected. In Example 3, when the pH is adjusted to the appropriate value, the antioxidant and cell renewal and repair promoting effects of small molecule peptides are fully exerted. Compared with adjusting pH with triethanolamine alone, the product has a more prominent effect on improving the comprehensive skin condition.

[0078] Example 4

[0079] Composition formula

[0080] By weight, 25 parts of γ-polyglutamic acid, 12 parts of plant-derived small molecule peptide (amino acid sequence four), 35 parts of a mixed humectant of glycerol and butanediol (1:1), 3 parts of a mixed thickener of xanthan gum and carbomer (1:1), 1.8 parts of a mixed preservative of phenoxyethanol and methyl paraben (1:1), 0.6 parts of a mixed pH regulator of citric acid and sodium citrate (1:1), and the balance is deionized water.

[0081] Preparation process

[0082] 1. Preliminary selection of plant-derived small molecule peptides: Collect plant samples in desert areas, and after screening, Opuntia stricta is determined as the extraction object.

[0083] 2. Extraction and purification of plant-derived small molecule peptides: Wash, dry, and crush Opuntia stricta, and use supercritical carbon dioxide extraction with an extraction pressure of 28 MPa and a temperature of 43 °C, and use 58% ethanol as the entrainer for extraction for 2.8 hours. The crude extract is purified by a macroporous adsorption resin column, ultrafiltered, and freeze-dried to obtain small molecule peptides.

[0084] 3. Sequencing of plant-derived small molecule peptides: Using LC-MS / MS technology combined with Mascot software, the amino acid sequence of the small molecule peptide is determined to be amino acid sequence four.

[0085] 4. Pretreatment of γ-polyglutamic acid: Prepare a 17% solution of γ-polyglutamic acid, remove impurities through an ion exchange resin, and concentrate it under reduced pressure to 37% at 52 °C for standby.

[0086] 5. Preparation of the composition: Add 80% deionized water to the reaction kettle, add the mixed thickener and swell for 1.4 hours while stirring at 170 r / min, sequentially add the mixed humectant, γ-polyglutamic acid solution, and small molecule peptide, stir for 46 minutes, add the mixed preservative and stir for 23 minutes,

[0087] Adjust the pH to 5.6 with the mixed pH regulator, and make up the volume and stir evenly.

[0088] Quality inspection: The product has a uniform and delicate appearance, no peculiar smell, a pH value of 5.6, good viscosity, no pathogenic bacteria detected in microorganisms, and the content of active ingredients meets the standards. Comparison with the sole use of γ-polyglutamic acid: The sole use of γ-polyglutamic acid has a certain effect in aspects such as moisturization. The small molecule peptide of amino acid sequence four added in Example 4 contains amino acids such as hydroxyproline (Hyp) that promote collagen synthesis. In synergy with γ-polyglutamic acid, γ-polyglutamic acid maintains the moisture on the skin surface, and the small molecule peptide promotes collagen synthesis to increase skin elasticity and firmness. In the volunteer trial, the product of Example 4 makes the skin elasticity increase more significantly, reduces wrinkles, and the overall firmness is better than that of the sole use of γ-polyglutamic acid.

[0089] 1. Comparison with the sole use of the mixed humectant of glycerol and butanediol: When the mixed humectant of glycerol and butanediol is used alone, it constructs a moisture retention network. In Example 4, the small molecule peptide synergizes with the mixed humectant to regulate cellular aquaporins and promote the absorption and utilization of the moisture maintained by the mixed humectant by cells. The strong moisture retention ability of new cells and the mixed humectant jointly reinforce the moisture retention system. In the skin moisture loss experiment, the moisture loss rate of the product sample of Example 4 is much lower than that of the sample using only the mixed humectant, and the moisturizing effect is greatly improved.

[0090] 2. Comparison with the sole use of the mixed thickener of xanthan gum and carbomer: The sole use of the mixed thickener of xanthan gum and carbomer improves the consistency and spreadability of the product. In Example 4, the mixed thickener synergizes with the small molecule peptide to stabilize the system and make the small molecule peptide evenly dispersed. The good spreadability increases the contact between the small molecule peptide and the skin. Consumer feedback shows that the product of Example 4 has a smooth spread, and its effect in promoting collagen synthesis and improving skin elasticity and firmness is better than that of the product containing only the mixed thickener, highlighting the promoting effect of synergy on the efficacy of the small molecule peptide.

[0091] Comparison with the sole use of the mixed preservatives of phenoxyethanol and methylparaben: The mixed preservatives inhibit bacteria when used alone. In Example 4, the antioxidant ability of the small molecule peptide assists the mixed preservatives in inhibiting microorganisms. Comparison with the sole use of the mixed preservatives of phenoxyethanol and methylparaben: The mixed preservatives inhibit bacteria when used alone. In Example 4, the antioxidant ability of the small molecule peptide assists the mixed preservatives in inhibiting microorganisms. The environment created by the mixed preservatives maintains the activity of the small molecule peptide. In the product microbial stability test, the retention rate of the activity of the small molecule peptide in the product of Example 4 is higher, and both the product quality and the activity of the small molecule peptide are better maintained. After being placed for 3 months in an environment simulating actual use, the activity of the small molecule peptide in the product using only the mixed preservatives drops to 65%, while the activity of the small molecule peptide in the product of Example 4 still remains at 82%, and the microbial indicators are always qualified, indicating that the synergy between the two effectively guarantees the stability and efficacy of the product.

[0092] 1. Comparison with the use of the mixed pH regulator of citric acid and sodium citrate alone: The mixed pH

[0093] regulator alone adjusts the pH. In Example 4, the pH is adjusted to the appropriate range, and the effects such as promoting collagen synthesis by small molecular peptides are fully exerted. Compared with the use of the mixed pH regulator alone, the product has better effects in improving skin elasticity and firmness, etc., reflecting the enhancing effect of the appropriate pH environment on the efficacy of small molecular peptides. In in vitro cell experiments, when the pH of the cell culture medium is adjusted to an inappropriate range with the mixed pH regulator, the promotion rate of small molecular peptides on collagen synthesis is only 30%; while under the appropriate pH conditions of Example 4, the promotion rate of small molecular peptides on collagen synthesis reaches 55%, indicating that the appropriate pH can significantly enhance the efficacy of small molecular peptides.

[0094] Summary of comprehensive comparison and synergistic effect

[0095] In terms of moisturizing

[0096] From the four examples, γ-polyglutamic acid synergistically acts with small molecular peptides extracted from plants of different sources and various moisturizers, significantly improving the moisturizing effect of the product. In Example 1, glycerol is combined with small molecular peptides from Sargassum, and it performs better than glycerol alone in the skin moisture loss test; in Example 2, butylene glycol synergistically acts with small molecular peptides from Rhodiola rosea, strengthening the moisturizing system; in Example 3, sodium hyaluronate and small molecular peptides from Undaria pinnatifida make the skin's water retention ability stronger; in Example 4, the mixed moisturizer of glycerol and butylene glycol and small molecular peptides from Opuntia ficus-indica greatly improves the moisturizing effect. Generally speaking, these combinations not only increase the water content on the skin surface in terms of moisturizing, but also promote the uptake and retention of water by cells, strengthening the moisturizing effect from different levels, far exceeding the use of each component alone.

[0097] In terms of skin repair and improvement

[0098] Small molecular peptides from different plant sources bring diverse skin repair and improvement effects to the product. In Example 1, small molecular peptides from Sargassum promote collagen synthesis and enhance skin elasticity; in Example 2, small molecular peptides from Rhodiola rosea have antioxidant and skin pH balance maintaining effects; in Example 3, small molecular peptides from Undaria pinnatifida improve skin color and texture; in Example 4, small molecular peptides from Opuntia ficus-indica increase skin elasticity and firmness and reduce wrinkles. After synergistically acting with γ-polyglutamic acid, in volunteer trials and experimental tests, these combinations have significant effects in comprehensive skin improvement, and have better performance than γ-polyglutamic acid alone in terms of moisturizing, elasticity, color, etc.

[0099] In terms of product stability

[0100] Under the synergistic effect of preservatives and small molecule peptides, both the microbial stability of the product and the retention rate of small molecule peptide activity are improved. In Example 1, phenoxyethanol and small molecule peptides; in Example 2, methylparaben and small molecule peptides; in Example 3, potassium sorbate and small molecule peptides; in Example 4, a mixed preservative of phenoxyethanol and methylparaben and small molecule peptides all demonstrate that the antioxidant ability of small molecule peptides assists the bacteriostatic effect of preservatives, and the preservatives maintain the activity of small molecule peptides in the environment, making the quality of the product more stable during storage and use.

[0101] In terms of pH adjustment

[0102] Appropriate pH regulators can ensure that small molecule peptides are in the best active state, thus giving full play to their efficacy. In Example 1, citric acid; in Example 2, sodium citrate; in Example 3, triethanolamine; in Example 4, a mixed pH regulator of citric acid and sodium citrate, after adjusting the pH of the product to the appropriate range, all significantly enhance the efficacy of small molecule peptides in promoting collagen synthesis, antioxidant and other aspects, which is better than the performance of small molecule peptides when using pH regulators alone.

[0103] In summary, through the synergistic effect of small molecule peptides from different plant sources and other components, significant comparative synergistic advantages have been demonstrated in aspects such as moisturizing, skin repair, product stability and pH adjustment, providing more effective formulation ideas and practical bases for the development and application of beauty products.

[0104] Comparative Example 1

[0105] Composition formula

[0106] By weight, 10 parts of γ-polyglutamic acid, 5 parts of small molecule peptides with common amino acid sequences (commercially available), 30 parts of glycerin, 2 parts of xanthan gum, 1 part of phenoxyethanol, 0.2 part of citric acid, and the balance is deionized water.

[0107] Preparation process

[0108] The preparation process is the same as that of Example 1, only replacing the plant-extracted small molecule peptides with commercially available common small molecule peptides.

[0109] Quality inspection and effect comparison

[0110] The product passes the quality inspection. However, in the efficacy test, through the comparison of volunteer trials, the volunteers using the product of Example 1 of the present invention have significantly better effects in terms of skin moisturizing and firmness improvement than the volunteers using the product of Comparative Example 1, indicating that the plant-extracted small molecule peptides of the present invention have better efficacy.

[0111] Comparative Example 2

[0112] Composition formula

[0113] By weight, 10 parts of γ-polyglutamic acid, 5 parts of small molecule peptides extracted from plants, 30 parts of glycerol, 2 parts of xanthan gum, 1 part of phenoxyethanol, 0.2 part of citric acid, and the balance is deionized water.

[0114] Preparation process

[0115] The small molecule peptides extracted from plants are extracted by the traditional organic solvent extraction method, and the other steps are the same as those in Example 1.

[0116] Quality inspection and effect comparison

[0117] The product quality inspection is qualified, but in the stability test, the product of Comparative Example 2 shows slight stratification after 3 months of storage, while the product of Example 1 has no obvious change, indicating that the advanced preparation processes such as supercritical carbon dioxide extraction adopted in the present invention have a positive impact on the product stability.

[0118] Comparative synergistic analysis of small molecule peptides and each component in the beauty composition

[0119] In the composition of γ-polyglutamic acid and related small molecule peptides for beauty products of the present invention, each component does not exist in isolation, but synergizes with each other to jointly exert excellent effects. The following will elaborate on the comparative synergistic effects of small molecule peptides and other main components in detail.

[0120] Synergistic effect between small molecule peptides and γ-polyglutamic acid

[0121] Synergy of moisturizing and cell regulation

[0122] Due to its unique molecular structure, γ-polyglutamic acid has a large number of carboxyl groups densely distributed on its molecular chain, which can adsorb a large amount of water molecules, build a strong moisturizing barrier on the skin surface, significantly increase the water content of the skin stratum corneum, reduce water loss, and keep the skin moist at all times. However, its function is mainly concentrated in the moisturizing link on the skin surface layer.

[0123] In contrast, the small molecule peptides extracted from plants with a new amino acid sequence in the present invention, although their moisturizing ability is relatively weak, the special arrangement of amino acid residues contained in them, such as the Pro-His-Cys tandem structure in amino acid sequence one, can precisely bind to the receptors on the surface of skin cells. This binding triggers a series of complex signal transduction pathways in the cells, activating the key gene expressions related to cell metabolism and collagen synthesis. For example, it promotes fibroblasts to synthesize more collagen fibers, enhances the elasticity and firmness of the skin, and improves the skin state from the cellular level.

[0124] When combined, γ-polyglutamic acid maintains a moist environment on the skin surface, providing favorable conditions for small molecule peptides to smoothly penetrate the stratum corneum and reach the dermal layer cells; small molecule peptides, on the other hand, further consolidate the moisturizing effect of γ-polyglutamic acid by promoting cell metabolism and enhancing the skin's own moisturizing ability and repair function. In a 4-week volunteer trial, in the experimental group using a product containing a composition of γ-polyglutamic acid and small molecule peptides, the skin moisture content increased by 35%, and the skin elasticity increased by 28%; while in the control group using only γ-polyglutamic acid products, the skin moisture content increased by 20%, and the elasticity only increased by 10%; in the control group using only small molecule peptide products, the moisture content increased by 12%, and the elasticity increased by 15%. Thus, the synergistic effect of the two can comprehensively improve the skin's moisture and firmness, and the effect far exceeds that of using them alone.

[0125] Synergistic Enhancement of Small Molecule Peptides and Humectants

[0126] Strengthening of the Multifunctional Moisturizing Network

[0127] Common humectants such as glycerol, butanediol, sodium hyaluronate, etc. each have their own moisturizing mechanisms. Glycerol molecules contain three hydroxyl groups, can form a large number of hydrogen bonds with water molecules, have strong hygroscopicity, can absorb water from the surrounding environment and retain it on the skin surface; butanediol also relies on the hydroxyl groups in its molecular structure to assist glycerol in enhancing the moisturizing effect; sodium hyaluronate, due to its unique macromolecular structure, can bind thousands of times its own weight of water, forming a water-rich gel-like substance on the skin surface to exert an efficient moisturizing effect.

[0128] When small molecule peptides act synergistically with these humectants, they exhibit unique advantages. Small molecule peptides can regulate the expression of aquaporin in skin cells and promote the balanced transport of water inside and outside cells. On the one hand, it enhances the uptake and utilization efficiency of the water adsorbed by humectants by cells, enabling the moisturizing effect of humectants on the skin surface to be better transmitted to the cell interior; on the other hand, the new cells generated by small molecule peptides promoting cell metabolism have stronger water retention ability, echoing the moisturizing network constructed by humectants on the skin surface. In a skin water loss experiment, for the sample using a composition of small molecule peptides and humectants, after being placed in a dry environment for 8 hours, the water loss rate was only 18%; while for the sample using only humectants, the water loss rate reached 30%. This shows that the synergy of small molecule peptides and humectants significantly strengthens the skin's moisturizing barrier and improves the durability and effect of moisturization.

[0129] Synergistic Enhancement of Small Molecule Peptides and Thickening Agents

[0130] Optimization of Skin Feel and Efficacy Carrier

[0131] Thickeners such as xanthan gum, carbomer, hydroxyethyl cellulose, etc. The main function is to regulate the rheological properties of the composition, giving the product an appropriate consistency and good spreadability. For example, carbomer can form a three-dimensional network structure in aqueous solution, making the product have a certain viscosity and thixotropy, which is convenient for consumers to evenly apply on the skin.

[0132] When small molecule peptides are combined with thickeners, the stable system formed by the thickeners helps the small molecule peptides to be evenly dispersed in the product, preventing their aggregation and precipitation, and ensuring the stability of small molecule peptides during storage and use. At the same time, the improved spreadability of the product by the thickener enables the small molecule peptides to cover the skin surface more evenly, increasing the contact area with skin cells, thereby improving the absorption efficiency and efficacy of small molecule peptides. In the consumer use experience survey, 90% of the volunteers reported that the products containing small molecule peptides and thickener compositions were smooth to apply, and the skin improvement effect was more obvious after use; while for the products containing only thickeners without adding small molecule peptides, although the spreadability was good, the feedback on skin efficacy improvement was only 50%. This reflects the synergy between small molecule peptides and thickeners, which greatly promotes the efficacy of small molecule peptides while optimizing the skin feel of product use.

[0133] Synergistic effect of small molecule peptides and preservatives

[0134] Activity protection and safety guarantee

[0135] Preservatives such as phenoxyethanol, methyl paraben, potassium sorbate, etc. play a crucial antibacterial role in beauty products, preventing the growth of microorganisms and ensuring the quality and safety of products within the shelf life.

[0136] When small molecule peptides act synergistically with preservatives, on the one hand, small molecule peptides themselves have a certain antioxidant capacity, which can reduce the deterioration of product components caused by oxidation, lower the risk of microorganism growth, and assist preservatives to better play their antibacterial effects; on the other hand, the sterile environment created by preservatives is conducive to maintaining the biological activity of small molecule peptides, preventing them from being degraded by microorganisms or inactivated due to the influence of microbial metabolites. In the product microbial stability test, the products containing small molecule peptides and preservative compositions still met the standards in terms of microbial indicators after being stored under accelerated aging conditions for 6 months, and the retention rate of small molecule peptide activity reached 85%; while for the products containing only preservatives, the retention rate of small molecule peptide activity was only 60%. This shows that the synergy between small molecule peptides and preservatives can not only ensure the safety of products, but also effectively protect the activity of small molecule peptides and extend the effective use period of products.

[0137] Synergistic effect of small molecule peptides and pH regulators

[0138] Stable environment and efficacy promotion

[0139] pH regulators such as citric acid, sodium citrate, triethanolamine, etc. are used to precisely adjust the pH value of the composition to 5.0 - 7.0. This pH range matches the weakly acidic environment on the skin surface, which can reduce the irritation of the product to the skin and ensure the chemical stability of each component in the product.

[0140] The biological activity of small molecule peptides is relatively sensitive to the environmental pH value. Under suitable pH conditions, their molecular structure can remain stable and their binding ability to skin cell receptors is stronger. The stable pH environment created by the pH regulator ensures that small molecule peptides are always in the best active state, giving full play to their effects of promoting cell metabolism and collagen synthesis. In in vitro cell experiments, when the pH value was maintained at 5.5 - 6.5, the expression level of genes related to collagen synthesis in the cell culture medium with small molecule peptides added was increased by 3 - 5 times compared with when the pH value deviated from this range. This fully demonstrates the synergy between the pH regulator and small molecule peptides, creating an ideal chemical environment for the efficacy of small molecule peptides and greatly enhancing the efficacy of small molecule peptides in beauty products.

[0141] From the above examples and comparative examples, it can be seen that the composition of γ-polyglutamic acid and related small molecule peptides for beauty products and its preparation process of the present invention have innovation and superiority, can prepare beauty products with significant efficacy and stable quality, and have good market application prospects.

[0142] Comparative Example 3

[0143] Analysis of the synergistic effect of Example 1

[0144] 1. Comparison with the use of γ-polyglutamic acid alone: When γ-polyglutamic acid is used alone, it mainly forms a moisturizing barrier on the skin surface and improves the water content of the skin stratum corneum. However, in the 4-week volunteer trial experiment, in the control group using γ-polyglutamic acid product alone, the skin moisture content increased by 20%, and the elasticity only increased by 10%. After adding small molecule peptides with specific amino acid sequence 1 in Example 1, the skin moisture content of the experimental group increased by 35%, and the skin elasticity increased by 28%. This is because small molecule peptides can activate the signal pathway promoting collagen synthesis in cells, promote fibroblasts to synthesize more collagen fibers, enhance skin elasticity, and at the same time their role in promoting cell metabolism, in turn, enhances the skin's own moisturizing ability, further consolidating the moisturizing effect of γ-polyglutamic acid, comprehensively improving the skin condition, and the effect far exceeds the use of γ-polyglutamic acid alone.

[0145] 2. Comparison with the use of glycerol alone: When glycerol is used alone, it mainly relies on the three hydroxyl groups in the molecule to form hydrogen bonds with water molecules, absorbing moisture from the surrounding environment and retaining it on the skin surface. In the skin moisture loss experiment, after the sample containing only glycerol was placed in a dry environment for 8 hours, the moisture loss rate reached 30%. When glycerol and small molecule peptides act synergistically in Example 1, the small molecule peptides regulate the expression of aquaporins in skin cells, promote the balanced transport of water inside and outside the cells, enhance the uptake and utilization efficiency of the water adsorbed by glycerol by the cells, enable the moisturizing effect of glycerol on the skin surface to be better transmitted to the interior of the cells. At the same time, the new cells generated by the promotion of cell metabolism by small molecule peptides have stronger water retention ability, echoing the moisturizing network constructed by glycerol. The moisture loss rate of the sample containing the composition of small molecule peptides and glycerol is only 18%, significantly strengthening the skin's moisturizing barrier and improving the durability and effect of moisturization.

[0146] 3. Comparison with the use of xanthan gum alone: When xanthan gum is used alone, it can regulate the rheological properties of the product, giving the product an appropriate consistency and certain spreadability. However, in the consumer experience survey, for the product containing only xanthan gum without small molecule peptides, although the spreadability is good, the feedback on skin efficacy improvement is only 50%. When xanthan gum and small molecule peptides are combined in Example 1, the stable system formed by xanthan gum helps the small molecule peptides to be evenly dispersed in the product, preventing their aggregation and precipitation, and ensuring the stability of small molecule peptides during storage and use. At the same time, the improved spreadability of the product by xanthan gum enables the small molecule peptides to cover the skin surface more evenly, increasing the contact area with skin cells, thereby improving the absorption efficiency and efficacy of small molecule peptides. 90% of the volunteers reported that the product containing the composition of small molecule peptides and xanthan gum has smooth spreading and more obvious skin improvement effect after use.

[0147] 4. Comparison with the use of phenoxyethanol alone: When phenoxyethanol is used alone, it can effectively inhibit the growth of common bacteria, molds and yeasts, preventing the product from being contaminated by microorganisms. In the product microbial stability test, for the product containing only phenoxyethanol, the retention rate of small molecule peptide activity is only 60%. When phenoxyethanol and small molecule peptides act synergistically in Example 1, the small molecule peptides themselves have certain antioxidant ability, which can reduce the deterioration of product components caused by oxidation, reduce the risk of microbial growth, and assist phenoxyethanol to better exert its antibacterial effect. At the same time, the sterile environment created by phenoxyethanol is conducive to maintaining the biological activity of small molecule peptides, preventing them from being degraded by microorganisms or inactivated by the influence of microbial metabolites. After the product containing the composition of small molecule peptides and phenoxyethanol is stored under accelerated aging conditions for 6 months, the microbial indicators still meet the standards, and the retention rate of small molecule peptide activity reaches 85%.

[0148] 5. Comparison with the pH adjustment by citric acid alone: Citric acid alone can adjust the pH value of the product, but the biological activity of small molecule peptides is relatively sensitive to the environmental pH value. In in vitro cell experiments, when the pH value is adjusted only by citric acid and deviates from the range of 5.5 - 6.5, the expression level of genes related to collagen synthesis in the cell culture medium added with small molecule peptides increases limitedly. In Example 1, citric acid is used to adjust the pH to 6.0, which is within the pH range suitable for the activity of small molecule peptides, ensuring that the small molecule peptides are always in the best active state and fully exerting their effects of promoting cell metabolism and collagen synthesis. Compared with adjusting the pH with citric acid alone, the efficacy of small molecule peptides in beauty products is greatly improved.

[0149] Comparative Synergistic Analysis of Example 2

[0150] 1. Comparison with the use of γ-polyglutamic acid alone: In the control group using γ-polyglutamic acid product alone, the effects on improving skin moisture and elasticity are limited. In Example 2, due to the addition of small molecule peptides with amino acid sequence II, tryptophan (Trp) and phenylalanine (Phe) in its sequence have antioxidant properties, and lysine (Lys), arginine (Arg), etc. contribute to maintaining the skin's acid-base balance and synergize with γ-polyglutamic acid. γ-polyglutamic acid provides surface moisturization, and small molecule peptides improve the skin at the cellular level. In the volunteer trial, the product of Example 2 has a higher increase in skin moisture content than the group using γ-polyglutamic acid alone, and performs better in aspects such as improving the skin's resistance to free radical damage and maintaining acid-base balance, with significantly improved comprehensive effects.

[0151] 2. Comparison with the use of butanediol alone: When butanediol acts alone, it assists in moisturization through the hydroxyl groups in the molecule. In Example 2, small molecule peptides and butanediol synergize. Small molecule peptides regulate the expression of aquaporin in cells, making the skin surface moisture maintained by butanediol uptake more efficient by cells, and enhancing the water retention ability of newly generated cells, jointly constructing a more stable moisturizing system. In the skin moisture loss test, the sample moisture loss rate of the product of Example 2 is significantly lower than that of the sample using only butanediol, and the moisturizing effect is greatly enhanced.

[0152] 3. Comparison with the use of carbomer alone: Carbomer alone can improve the spreadability of the product. In Example 2, carbomer and small molecule peptides synergize. The stable system allows small molecule peptides to be evenly dispersed, and the good spreadability increases the contact between small molecule peptides and skin cells. Consumers feedback that using the product of Example 2 is not only smooth to apply, but also significantly superior to the product containing only carbomer in terms of antioxidant and maintaining skin acid-base balance effects, indicating that small molecule peptides play a better role by virtue of the characteristics of carbomer.

[0153] 4. Comparison with the use of methyl p-hydroxybenzoate alone: When methyl p-hydroxybenzoate is used alone for antibacterial purposes, the activity of small molecule peptides is easily affected. In Example 2, the antioxidant property of small molecule peptides reduces the risk of microbial growth, assisting methyl p-hydroxybenzoate in antibacterial action. Meanwhile, the antibacterial environment safeguards the activity of small molecule peptides. In the product microbial stability test, the retention rate of the activity of small molecule peptides in the product of Example 2 is higher than that of the product containing only methyl p-hydroxybenzoate, and both the product quality and the activity of small molecule peptides are better guaranteed.

[0154] 5. Comparison with the use of sodium citrate alone for pH adjustment: When sodium citrate is used alone to adjust the pH, if the pH is not appropriate, the activity of small molecule peptides is limited. In Example 2, the pH is adjusted to 5.5, which is within the optimal range of the activity of small molecule peptides. The antioxidant and skin pH balance maintenance effects of small molecule peptides are fully exerted. Compared with the use of sodium citrate alone for pH adjustment, the product has a better effect in improving the skin condition.

[0155] Comparative Synergistic Analysis of Example 3

[0156] 1. Comparison with the use of γ-polyglutamic acid alone: When γ-polyglutamic acid is used alone, it mainly focuses on skin moisturization. In Example 3, the small molecule peptide with amino acid sequence three is added. The sulfhydryl group of cysteine (Cys) in it participates in antioxidant defense and regulates cell gene expression to promote renewal and repair. After synergizing with γ-polyglutamic acid, not only is the moisturizing effect enhanced due to the small molecule peptide promoting cell metabolism, but also better effects are achieved in improving skin color and texture. In the feedback from volunteers, the product of Example 3 makes the skin more moisturized, with a more uniform color and a finer texture, which is better than the effect of using γ-polyglutamic acid alone.

[0157] 2. Comparison with the use of sodium hyaluronate alone: When sodium hyaluronate is used alone, it can bind a large amount of water to form a moisturizing gel on the skin surface. In Example 3, the small molecule peptide synergizes with sodium hyaluronate. The small molecule peptide regulates the absorption and utilization of water by cells, enabling the moisturizing effect of sodium hyaluronate to penetrate better into the cell level. At the same time, the new cells generated by promoting cell metabolism and sodium hyaluronate jointly strengthen the moisturizing network. In the skin moisture content test, the product of Example 3 has a stronger skin moisture retention ability and a more lasting moisturizing effect.

[0158] 3. Comparison with the use of hydroxyethyl cellulose alone: When hydroxyethyl cellulose is used alone, it can adjust the rheological properties of the product. In Example 3, hydroxyethyl cellulose synergizes with the small molecule peptide to ensure the uniform distribution of the small molecule peptide. Its improved spreadability helps the small molecule peptide to come into contact with skin cells more comprehensively. User experience shows that when using the product of Example 3, the application process is comfortable, and the effects in antioxidant and improving skin color and texture are significantly better than those of the product containing only hydroxyethyl cellulose, reflecting the promotion of the synergistic effect of the two on the efficacy of small molecule peptides.

[0159] 4. Comparison with the use of potassium sorbate alone: When potassium sorbate is used alone for antibacterial purposes, the activity of small molecule peptides may be impaired. In Example 3, the antioxidant ability of small molecule peptides aids potassium sorbate in inhibiting microbial growth, and the sterile environment created by potassium sorbate maintains the activity of small molecule peptides. In the product microbial stability test, the retention rate of small molecule peptide activity in the product of Example 3 is higher, ensuring the quality and small molecule peptide activity during the product's shelf life.

[0160] 5. Comparison with the use of triethanolamine alone to adjust pH: If the pH adjusted by triethanolamine alone does not reach the appropriate range, the activity of small molecule peptides will be affected. In Example 3, the pH is adjusted to the appropriate value, enabling the full exertion of the antioxidant and cell renewal and repair-promoting effects of small molecule peptides. Compared with the use of triethanolamine alone to adjust pH, the product shows a more prominent effect in improving the overall skin condition.

[0161] Comparative Synergistic Analysis of Example 4

[0162] 1. Comparison with the use of γ-polyglutamic acid alone: The use of γ-polyglutamic acid alone has certain effects in aspects such as moisturization. The small molecule peptide with amino acid sequence four added in Example 4 contains amino acids such as hydroxyproline (Hyp) that promote collagen synthesis. In synergy with γ-polyglutamic acid, γ-polyglutamic acid maintains the moisture on the skin surface, and the small molecule peptide promotes collagen synthesis to increase skin elasticity and firmness. In the volunteer trial, the product of Example 4 shows a more significant improvement in skin elasticity, reduced wrinkles, and overall better firmness compared to the use of γ-polyglutamic acid alone.

[0163] 2. Comparison with the use of a mixture of glycerol and butanediol as a humectant alone: When a mixture of glycerol and butanediol is used alone as a humectant, it constructs a moisture retention network. In Example 4, the small molecule peptide synergizes with the mixed humectant to regulate aquaporins in cells, promoting the absorption and utilization of the moisture maintained by the mixed humectant by cells. The strong moisture retention ability of new cells and the mixed humectant jointly strengthen the moisture retention system. In the skin moisture loss experiment, the moisture loss rate of the product sample of Example 4 is much lower than that of the sample using only the mixed humectant, significantly improving the moisturizing effect.

[0164] 3. Comparison with the use of a mixture of xanthan gum and carbomer as a thickener alone: The use of a mixture of xanthan gum and carbomer as a thickener alone improves the consistency and spreadability of the product. In Example 4, the mixed thickener synergizes with the small molecule peptide to stabilize the system, enabling the small molecule peptide to be evenly dispersed. The good spreadability increases the contact between the small molecule peptide and the skin. Consumer feedback indicates that the product of Example 4 spreads smoothly and shows better effects in promoting collagen synthesis and improving skin elasticity and firmness compared to the product containing only the mixed thickener, highlighting the promoting effect of synergy on the efficacy of small molecule peptides.

[0165] 4. Comparison with the use of the mixed preservatives of phenoxyethanol and methylparaben alone: The mixed preservatives alone have antibacterial properties. In Example 4, the antioxidant ability of the small molecule peptide assists the mixed preservatives in inhibiting microorganisms, and the environment created by the mixed preservatives maintains the activity of the small molecule peptide. In the product microbial stability test, the retention rate of the small molecule peptide activity in the product of Example 4 is higher, and both the product quality and the small molecule peptide activity are better maintained.

[0166] 5. Comparison with the use of the mixed pH regulator of citric acid and sodium citrate alone: The mixed pH regulator alone adjusts the pH. In Example 4, the pH is adjusted to the appropriate range, and the effects of the small molecule peptide in promoting collagen synthesis and other functions are fully exerted. Compared with the use of the mixed pH regulator alone, the product has better effects in improving skin elasticity and firmness, etc., reflecting the enhancing effect of the appropriate pH environment on the efficacy of the small molecule peptide.

[0167] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, and improvements made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A composition of γ-polyglutamic acid and related small molecule peptides for beauty products, characterized in that, By weight, it comprises the following components: 5-50 parts of γ-polyglutamic acid, 1-25 parts of small molecule peptides extracted from plants, 20-150 parts of humectant, 1-15 parts of thickener, 1-5 parts of preservative, 0.1-2.5 parts of pH regulator, and the balance is deionized water.

2. The composition of γ-polyglutamic acid and related small molecule peptides for beauty products according to claim 1, characterized in that, The small molecule peptides extracted from plants have at least one of the following amino acid sequences: Amino acid sequence one: Ala-Gly-Ser-Tyr-Pro-His-Cys-Val-Gln-Thr-Leu-Asn; Amino acid sequence two: Leu-Ile-Phe-Lys-Asp-Glu-Trp-Met-Pro-Gly-Thr-Ser; Amino acid sequence three: Cys-Gln-Tyr-His-Asn-Val-Leu-Ala-Pro-Ser-Thr-Gly; Amino acid sequence four: Gly-Pro-Hyp-Lys-Arg-Ser-Asp-Glu-Thr-Ala-Val-Ile.

3. The composition of γ-polyglutamic acid and related small molecule peptides for beauty products according to claim 1, characterized in that, The γ-polyglutamic acid is a homopolymer with a molecular weight between 500 kDa and 1500 kDa, and is prepared by the microbial fermentation method.

4. The composition of γ-polyglutamic acid and related small molecule peptides for beauty products according to claim 1, wherein, The humectant is one or a combination of glycerol, butanediol, and sodium hyaluronate.

5. The composition of γ-polyglutamic acid and related small molecule peptides for beauty products according to claim 1, characterized in that The thickener is one or more of xanthan gum, carbomer, and hydroxyethyl cellulose.

6. The composition of γ-polyglutamic acid and related small molecule peptides for beauty products according to claim 1, characterized in that, The preservative is one or more of phenoxyethanol, methyl paraben, and potassium sorbate.

7. The composition of γ-polyglutamic acid and related small molecule peptides for beauty products according to claim 1, characterized in that, The pH regulator is one or more of citric acid, sodium citrate, and triethanolamine, and the pH value of the composition is 5.0-7.

0.

8. A preparation process of a composition of γ-polyglutamic acid and related small molecule peptides for beauty products as described in any one of claims 1-7, characterized in that, It includes the following steps: Coarse selection of small molecule peptides extracted from plants: Collect plant samples with potential skin care effects in different ecological environments, establish a plant sample library, and select plants that may be rich in small molecule peptides with special biological activities based on traditional medicinal records, local folk use experience, and preliminary cell activity tests; Extraction and purification of small molecule peptides extracted from plants: Select specific plant raw materials determined by coarse selection, wash, dry, and then crush them into fine powder. Use supercritical carbon dioxide extraction technology for preliminary extraction, then purify through a macroporous adsorption resin column, and further remove impurities and macromolecular substances through ultrafiltration technology to obtain a high-purity solution of small molecule peptides extracted from plants. Freeze-dry for later use; Sequencing of small molecule peptides extracted from plants: Dissolve the freeze-dried small molecule peptide sample in an appropriate buffer solution, use liquid chromatography-mass spectrometry (LC-MS / MS) technology for sequencing analysis, and determine the amino acid sequence by comparing with a self-built database using bioinformatics software; Pretreatment of γ-polyglutamic acid: Dissolve the γ-polyglutamic acid powder prepared by the microbial fermentation method in deionized water to prepare a solution with a mass concentration of 12%-18%. Remove the impurity ions therein through an ion exchange resin column, and then perform vacuum concentration until the mass concentration of the solution reaches 32%-38% for later use; Preparation of the composition: Add 80% of the total amount of deionized water into a reaction kettle equipped with a stirring device, start stirring, slowly add the thickener, and stir evenly to make it fully swell. Then, sequentially add the humectant, the pretreated γ-polyglutamic acid solution, and the small molecule peptide extracted from plants, and continue stirring to fully mix the components. Next, add the preservative, adjust the pH value of the composition to 5.0 - 7.0 with a pH regulator after stirring, and finally add the remaining 20% of deionized water for volume fixation and stir evenly; Quality inspection: Conduct inspection items such as appearance, odor, pH value, viscosity, microbial index, and active ingredient content on the prepared composition, and package and store the qualified products.

9. The preparation process according to claim 8, characterized in that, The extraction pressure of the supercritical carbon dioxide extraction technology is 25 - 35 MPa, the extraction temperature is 40 - 50 °C, and ethanol (concentration 55% - 65%) is used as an entrainer to assist extraction for 2.5 - 3.5 hours.

10. The preparation process according to claim 8, characterized in that, The temperature of the vacuum concentration is controlled at 45 - 55 °C.

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