Extraction process of polypeptide rich in cannysia and repairing mask composition

Abstract

This invention relates to the field of skincare technology, and in particular to an extraction process for silkworm pupae polypeptides and a repair mask composition. The extraction process includes: defatting silkworm pupae powder using supercritical CO2 extraction to obtain defatted silkworm pupae powder; mixing the defatted silkworm pupae powder, a eutectic solvent, and an antioxidant, and then performing a cell-wall breaking process to obtain a cell-wall broken suspension; adding a complex enzyme to the cell-wall broken suspension and sequentially performing enzymatic hydrolysis, enzyme inactivation, and centrifugation to obtain a supernatant; the complex enzyme includes papain, bromelain, and neutral protease; filtering the supernatant once through a 30kDa filter membrane, collecting the filtrate, and then filtering it a second time through a 10kDa filter membrane to obtain the silkworm pupae polypeptide extract. The silkworm pupae polypeptide extract obtained by the extraction process of this invention has excellent DPPH free radical scavenging rate and no solvent residue, and can be directly used as a component of a repair mask, suitable for sensitive skin and skin damaged after cosmetic procedures.

Description

Technical Field

[0001] This invention relates to the field of skincare technology, and in particular to an extraction process for a product rich in silkworm peptides and a repair mask composition. Background Technology

[0002] Silkworm larvae (also known as Bombyx mori or Bombyx mori Linnaeus) are a commonly used traditional Chinese medicine for relieving wind and spasms. They are the dried bodies of the 4th-5th instar larvae of the silkworm (Bombyx mori Linnaeus), which die after natural infection or artificial inoculation with Beauveria bassiana (Bals.) Vuillant. Modern research shows that silkworm larvae are rich in protein components, ranging from 29.74% to 64.73%. Among these, polypeptide active ingredients are the core substances responsible for their anti-inflammatory, soothing, skin barrier repair, redness reduction, and antioxidant effects. They are well-suited as a core component in sensitive skin repair and post-medical aesthetic treatment soothing and repair masks, demonstrating significant application potential.

[0003] However, traditional peptide extraction processes have significant limitations: high-temperature extraction directly destroys the spatial structure of peptides, leading to the inactivation of core active ingredients and loss of their original skincare efficacy; while organic solvent extraction can improve extraction efficiency, it is prone to leaving organic reagent residues, which can aggravate skin irritation when used as cosmetic raw materials, failing to meet the requirements of gentleness and low allergenicity for sensitive skin products, thus restricting the large-scale application of silkworm peptides in the skincare field. Summary of the Invention

[0004] The purpose of this invention is to provide an extraction process and a repair mask composition rich in silkworm peptides, so as to solve the defects of existing silkworm peptide extraction processes, such as low peptide yield, large activity loss, and solvent residue in the final product, which makes it difficult to meet the requirements of skin care products for gentleness and low allergenicity.

[0005] To achieve the above objectives, the present invention provides an extraction process rich in silkworm peptides, comprising the following steps: S1. The silkworm powder was degreased by supercritical CO2 extraction to obtain degreased silkworm powder; S2. Mix defatted silkworm powder, eutectic solvent, and antioxidant, and then perform cell wall disruption treatment to obtain a cell wall disruption suspension. S3. Add the complex enzyme to the cell wall disruption suspension and perform enzymatic hydrolysis, enzyme inactivation, and centrifugation sequentially to obtain the supernatant; the complex enzyme includes papain, bromelain, and neutral protease. S4. After filtering the supernatant once through a 30kDa filter membrane, collect the filtrate and then filter it a second time through a 10kDa filter membrane to obtain the silkworm polypeptide extract.

[0006] In this invention, the supercritical CO2 extraction temperature in step S1 is preferably 30-40℃, more preferably 32-38℃, and even more preferably 34-36℃; the pressure is preferably 20-25 MPa, more preferably 21-24 MPa, and even more preferably 22-23 MPa; the time is preferably 1.5-2 h; and the CO2 flow rate is preferably 20-25 L / h, more preferably 21-24 L / h, and even more preferably 22-23 L / h. Within this parameter range, the CO2 fluid has optimal solubility for oils, and the moderate flow rate ensures thorough degreasing while avoiding fluidization loss of the silkworm powder. After degreasing, vacuum separation is performed to obtain degreased silkworm powder free of oil and solvent residue.

[0007] In this invention, the silkworm powder has a mesh size of 200-300 mesh, and the preparation process includes: washing the silkworms and drying them until the moisture content is ≤6%, then pulverizing them through a 200-300 mesh sieve to obtain silkworm powder.

[0008] In this invention, the mass ratio of defatted silkworm powder, eutectic solvent and antioxidant in S2 is preferably 1:10-15:0.05-0.1, more preferably 1:11-14:0.06-0.09, and even more preferably 1:12-13:0.07-0.08.

[0009] In this invention, the eutectic solvent in S2 is composed of choline chloride and glycerol, and the molar ratio of choline chloride to glycerol is preferably 1:1.5-2, more preferably 1:1.6-1.8, and even more preferably 1:1.7. Choline chloride and glycerol have no skin irritation and are suitable for cosmetic applications, avoiding skin allergies caused by subsequent organic solvent residues.

[0010] In this invention, the antioxidant is vitamin C, which can inhibit the oxidative degradation of peptides.

[0011] In this invention, the power of the cell wall breaking process in S2 is preferably 80-120W, more preferably 90-110W, and even more preferably 100W; the time is preferably 3-5min, more preferably 3.5-4.5min, and even more preferably 4min; and the temperature is preferably 25-35℃, more preferably 27-32℃, and even more preferably 30℃.

[0012] In this invention, the mass ratio of the compound enzyme to defatted silkworm powder in S3 is preferably 2-5:100, more preferably 3-4:100, and even more preferably 3.5:100; the mass ratio of papain, bromelain, and neutral protease is preferably 1-2:1:1, and even more preferably 1.5:1:1.

[0013] In this invention, the enzymatic hydrolysis temperature in step S3 is preferably 45-50℃, more preferably 47-49℃, and even more preferably 48℃; the time is preferably 3-4h, and even more preferably 3.5h; the pH is preferably 6-7, and even more preferably 6.5; the enzymatic hydrolysis process is also accompanied by intermittent ultrasonic treatment, which is performed as follows: after enzymatic hydrolysis for 25-30min, ultrasonic treatment is turned on for 3-5min at a power of 150-200W; intermittent ultrasonic treatment enhances mass transfer efficiency and improves the degree of enzymatic hydrolysis, while avoiding the local high temperature caused by continuous ultrasonication that leads to peptide inactivation; the enzyme inactivation temperature is preferably 80-85℃, more preferably 82-84℃, and even more preferably 83℃; the time is preferably 4-6min, more preferably 4.5-5.5min, and even more preferably 5min; the centrifugation speed is 4000-6000r / min, more preferably 4500-5500r / min, and even more preferably 5000r / min.

[0014] The present invention also provides a silkworm polypeptide extract obtained by the above-described extraction process rich in silkworm polypeptide.

[0015] The present invention also provides the application of the above-mentioned silkworm polypeptide extract in a repair mask.

[0016] The present invention also provides a repair mask composition, comprising, by weight (100%), 3%-6% of the above-mentioned silkworm polypeptide extract, 0.1%-0.5% of dipotassium glycyrrhizate, 0.05%-0.2% of 4-tert-butylcyclohexanol, 3%-8% of glycerin, 2%-5% of butylene glycol, 2%-5% of 1,3-propanediol, 0.1%-0.3% of sodium hyaluronate, 0.2%-0.4% of p-hydroxyacetophenone, 0.1%-0.2% of ethylhexylglycerin, and the remainder being water.

[0017] The present invention provides a repair mask composition, which, based on 100% by weight, preferably contains 4%-5% of the above-mentioned silkworm polypeptide extract, 0.2%-0.4% of dipotassium glycyrrhizate, 0.06%-0.08% of 4-tert-butylcyclohexanol, 4%-7% of glycerin, 3%-4% of butylene glycol, 3%-4% of 1,3-propanediol, 0.1%-0.2% of sodium hyaluronate, 0.2%-0.3% of p-hydroxyacetophenone, 0.1%-0.2% of ethylhexylglycerin, and the remainder is water.

[0018] The silkworm polypeptide extract prepared in this invention is the core ingredient in the repair mask composition. It has the advantages of high DPPH free radical scavenging rate and small molecule easy transdermal absorption. It can quickly penetrate the skin base, remove skin free radicals, reduce oxidative stress damage, and relieve skin redness, dryness and itching from the root. At the same time, it can help repair the damaged skin barrier, enhance skin resistance, and has no sensitizing macromolecules or solvent residues, making it gentle and non-irritating to fragile skin.

[0019] Dipotassium glycyrrhizate is a soothing and anti-inflammatory ingredient that can quickly inhibit skin inflammation and relieve capillary dilation. It specifically improves sensitive skin, redness, and burning. It works synergistically with silkworm peptides to reduce skin sensitization and enhance the overall soothing effect. The formula is gentle and non-irritating.

[0020] 4-Ter-butylcyclohexanol can quickly block the transmission of skin irritation signals, rapidly relieving discomfort such as dryness, itching, stinging, and redness. It is gentle and has no side effects. When combined with dipotassium glycyrrhizate, it achieves double soothing and redness reduction, making it suitable for highly sensitive skin and short-term repair after cosmetic procedures.

[0021] Glycerin, butylene glycol, and 1,3-propanediol are mild polyol moisturizers that are alcohol-free and non-irritating. They can form a breathable moisturizing film on the skin surface to relieve dryness in damaged skin and improve skin hydration and tolerance.

[0022] Sodium hyaluronate can form a water-locking film on the skin's surface, reducing moisture loss and filling the gaps between skin cells. It also helps repair the damaged skin barrier, improves the skin's ability to retain moisture and its elasticity. In synergy with polyol moisturizers, it achieves dual moisturizing on both the surface and deep layers, meeting the hydration and repair needs of dry and damaged skin.

[0023] p-Hydroxyacetophenone has both antibacterial and antioxidant properties, which can inhibit the growth of bacteria and mold in the mask composition and extend the product's shelf life. Ethylhexylglycerin can further enhance the antibacterial effect of the preservative system, while also helping active ingredients penetrate the skin and improving the absorption efficiency of the mask's active ingredients.

[0024] The present invention has the following beneficial effects: This invention provides an extraction process rich in silkworm pupae polypeptides, comprising the following steps: S1, defatting silkworm pupae powder using supercritical CO2 extraction to obtain defatted silkworm pupae powder; S2, mixing defatted silkworm pupae powder, a eutectic solvent, and an antioxidant, and performing cell wall disruption to obtain a cell wall disruption suspension; S3, adding a complex enzyme to the cell wall disruption suspension and sequentially performing enzymatic hydrolysis, enzyme inactivation, and centrifugation to obtain a supernatant; the complex enzyme includes papain, bromelain, and neutral protease; S4, filtering the supernatant once through a 30kDa filter membrane, collecting the filtrate, and then filtering it a second time through a 10kDa filter membrane to obtain the silkworm pupae polypeptide extract.

[0025] This invention uses supercritical CO2 extraction to degrease silkworm powder, breaking the barrier of oil on protein substrates and effectively overcoming the problems of deterioration and off-odor caused by oil oxidation in peptide products. At the same time, the low temperature does not damage the spatial active structure of proteins and peptides, and there are no organic reagent residues, ensuring the safety of the extract in cosmetics.

[0026] This invention utilizes a combination of papain, bromelain, and neutral protease, whose action sites are complementary, to directionally hydrolyze large protein molecules into small-molecule repair peptides that are easily absorbed by the skin. Combined with intermittent ultrasound assistance to enhance the contact mass transfer efficiency between the enzyme and the substrate, the proportion of small-molecule active peptides is significantly increased, thereby improving the activity retention rate.

[0027] This invention first uses a 30kDa filter membrane for coarse filtration, which efficiently traps and removes allergenic macromolecular foreign proteins from silkworm pupae, reducing the risk of skin redness and sensitization. Then, it uses a 10kDa filter membrane for secondary fine filtration, which further removes small and medium-sized ineffective impurities and precisely enriches small-molecule silkworm pupae polypeptides that are suitable for skin absorption. These polypeptides can be directly added to the repair mask without additional detoxification or impurity removal treatment.

[0028] This invention provides an extraction process for silkworm peptides that is low-temperature, environmentally friendly, and simple. The process steps are simple and controllable, and the equipment is suitable for industrial mass production. The prepared silkworm peptide extract has excellent stability and does not separate, deteriorate, or precipitate when added to a mask system. When combined with soothing mask components, it can quickly exert anti-inflammatory, redness-reducing, and skin barrier-repairing effects, completely filling the technological gap in existing silkworm peptides that cannot be adapted to sensitive skin cosmetics.

[0029] The silkworm polypeptide extract prepared in this invention is applied to the repair mask composition. There is no organic solvent residue or irritating substance introduced throughout the process. It is gentle on the skin and has no risk of causing allergies. It can be safely used on sensitive skin and skin damaged after medical aesthetic procedures. At the same time, the extract polypeptide activity is fully preserved and has high biological activity. It can quickly penetrate the skin to exert anti-inflammatory, redness-reducing, and barrier-repairing effects. It works synergistically with the soothing and moisturizing components in the mask. Under the premise of ensuring safety, it greatly improves the redness-reducing, repairing, and skin-stabilizing effects of the repair mask, achieving the dual advantages of gentle skin care and highly effective repair.

[0030] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0031] Figure 1 These are the DPPH scavenging rates of the silkworm polypeptide extracts in Examples 1-6 of this invention. Detailed Implementation

[0032] The present invention will be further described below with reference to the accompanying drawings and embodiments. Unless otherwise defined, the technical or scientific terms used in this invention should be understood in their ordinary sense by those skilled in the art. The features mentioned above or in the specific examples mentioned in this invention can be combined arbitrarily, and these specific embodiments are only used to illustrate the invention and are not intended to limit the scope of the invention.

[0033] Example 1 After cleaning the silkworm pupae, dry them until the moisture content is ≤6%, then crush them and pass them through a 200-mesh sieve to obtain silkworm pupae powder for later use. S1. The above-mentioned silkworm powder was degreased by supercritical CO2 extraction to obtain degreased silkworm powder; the equipment parameters were: temperature 30℃, pressure 20MPa, time 1.5h, CO2 flow rate 20L / h.

[0034] S2. Mix 1g of defatted silkworm powder, 10g of eutectic solvent (molar ratio of choline chloride and glycerol is 1:1.5), and 0.05g of vitamin C (antioxidant). At 25°C, process the mixture with 100W power for 3 minutes to obtain a cell-wall broken suspension.

[0035] S3. Add 0.05g of a compound enzyme (papain, bromelain, and neutral protease in a mass ratio of 1:1:1) to the cell wall-breaking suspension, adjust the pH to 6, and enzymatically hydrolyze at 45℃ for 3h (during enzymatic hydrolysis, sonicate for 5min every 25min, with a sonication power of 150W). Then inactivate the enzyme by incubating at 80℃ for 4min, and centrifuge at 5000rpm to obtain the supernatant.

[0036] S4. After filtering the supernatant once through a 30kDa filter membrane, collect the filtrate and then filter it a second time through a 10kDa filter membrane to obtain the silkworm polypeptide extract.

[0037] Example 2 After cleaning the silkworm pupae, dry them until the moisture content is ≤6%, then crush them and pass them through a 220-mesh sieve to obtain silkworm pupae powder for later use. S1. The above-mentioned silkworm powder was degreased by supercritical CO2 extraction to obtain degreased silkworm powder; the equipment parameters were: temperature 32℃, pressure 21MPa, time 1.5h, CO2 flow rate 21L / h.

[0038] S2. Mix 1g of defatted silkworm powder, 11g of eutectic solvent (molar ratio of choline chloride and glycerol is 1:2), and 0.07g of vitamin C (antioxidant). At 25°C, the mixture is subjected to cell wall disruption at 100W for 3.5 minutes to obtain a cell wall disruption suspension.

[0039] S3. Add 0.02g of a compound enzyme (papain, bromelain, and neutral protease in a mass ratio of 1.5:1:1) to the cell wall-breaking suspension, adjust the pH to 6, and enzymatically hydrolyze at 50℃ for 3h (during enzymatic hydrolysis, sonicate for 5min every 25min, with a sonication power of 150W). Then inactivate the enzyme by incubating at 85℃ for 4min, and centrifuge at 4500rpm to obtain the supernatant.

[0040] S4. After filtering the supernatant once through a 30kDa filter membrane, collect the filtrate and then filter it a second time through a 10kDa filter membrane to obtain the silkworm polypeptide extract.

[0041] Example 3 After cleaning the silkworm pupae, dry them until the moisture content is ≤6%, then crush them and pass them through a 260-mesh sieve to obtain silkworm pupae powder for later use. S1. The above-mentioned silkworm powder was degreased by supercritical CO2 extraction to obtain degreased silkworm powder; the equipment parameters were: temperature 34℃, pressure 22MPa, time 1.5h, CO2 flow rate 22L / h.

[0042] S2. Mix 1g of defatted silkworm powder, 14g of eutectic solvent (molar ratio of choline chloride and glycerol is 1:1.6), and 0.08g of vitamin C (antioxidant). At 27°C, the mixture is subjected to cell wall disruption at 90W for 5 minutes to obtain a cell wall disruption suspension.

[0043] S3. Add 0.04 g of a compound enzyme (papain, bromelain, and neutral protease in a mass ratio of 2:1:1) to the cell wall-breaking suspension, adjust the pH to 6.5, and enzymatically hydrolyze at 48℃ for 3.5 h (during enzymatic hydrolysis, sonicate for 5 min every 25 min, with a sonication power of 150 W). Then inactivate the enzyme by incubating at 83℃ for 6 min, and centrifuge at 6000 rpm to obtain the supernatant.

[0044] S4. After filtering the supernatant once through a 30kDa filter membrane, collect the filtrate and then filter it a second time through a 10kDa filter membrane to obtain the silkworm polypeptide extract.

[0045] Example 4 After cleaning the silkworm pupae, dry them until the moisture content is ≤6%, then crush them and pass them through a 300-mesh sieve to obtain silkworm pupae powder for later use. S1. The above-mentioned silkworm powder was degreased by supercritical CO2 extraction to obtain degreased silkworm powder; the equipment parameters were: temperature 36℃, pressure 23MPa, time 1.5h, CO2 flow rate 24L / h.

[0046] S2. Mix 1g of defatted silkworm powder, 12g of eutectic solvent (molar ratio of choline chloride and glycerol is 1:1.8), and 0.06g of vitamin C (antioxidant). At 32℃, the mixture is subjected to cell wall disruption at 110W for 4 minutes to obtain a cell wall disruption suspension.

[0047] S3. Add 0.035g of a compound enzyme (papain, bromelain, and neutral protease in a mass ratio of 2:1:1) to the cell wall-breaking suspension, adjust the pH to 7, and enzymatically hydrolyze at 49℃ for 4h (during enzymatic hydrolysis, sonicate for 5min every 25min, with a sonication power of 150W), then inactivate the enzyme by incubating at 85℃ for 4min, and then centrifuge at 5500rpm to obtain the supernatant.

[0048] S4. After filtering the supernatant once through a 30kDa filter membrane, collect the filtrate and then filter it a second time through a 10kDa filter membrane to obtain the silkworm polypeptide extract.

[0049] Example 5 After cleaning the silkworm pupae, dry them until the moisture content is ≤6%, then crush them and pass them through a 300-mesh sieve to obtain silkworm pupae powder for later use. S1. The above-mentioned silkworm powder was degreased by supercritical CO2 extraction to obtain degreased silkworm powder; the equipment parameters were: temperature 38℃, pressure 24MPa, time 2h, CO2 flow rate 23L / h.

[0050] S2. Mix 1g of defatted silkworm powder, 13g of eutectic solvent (molar ratio of choline chloride and glycerol is 1:2), and 0.09g of vitamin C (antioxidant). At 35°C, the mixture is subjected to cell wall disruption at 80W for 4.5 minutes to obtain a cell wall disruption suspension.

[0051] S3. Add 0.04g of a compound enzyme (papain, bromelain, and neutral protease in a mass ratio of 2:1:1) to the cell wall-breaking suspension, adjust the pH to 6, and enzymatically hydrolyze at 48℃ for 3h (during enzymatic hydrolysis, sonicate for 5min every 25min, with a sonication power of 150W). Then inactivate the enzyme by incubating at 82℃ for 6min, and centrifuge at 4500rpm to obtain the supernatant.

[0052] S4. After filtering the supernatant once through a 30kDa filter membrane, collect the filtrate and then filter it a second time through a 10kDa filter membrane to obtain the silkworm polypeptide extract.

[0053] Example 6 After cleaning the silkworm pupae, dry them until the moisture content is ≤6%, then crush them and pass them through a 200-mesh sieve to obtain silkworm pupae powder for later use. S1. The above-mentioned silkworm powder was degreased by supercritical CO2 extraction to obtain degreased silkworm powder; the equipment parameters were: temperature 40℃, pressure 25MPa, time 2h, CO2 flow rate 25L / h.

[0054] S2. Mix 1g of defatted silkworm powder, 15g of eutectic solvent (molar ratio of choline chloride and glycerol is 1:1.7), and 0.1g of vitamin C (antioxidant). At 30℃, the mixture is subjected to cell wall disruption at 120W for 3 minutes to obtain a cell wall disruption suspension.

[0055] S3. Add 0.03g of a compound enzyme (papain, bromelain, and neutral protease in a mass ratio of 1.5:1:1) to the cell wall-breaking suspension, adjust the pH to 6, and enzymatically hydrolyze at 50℃ for 3h (during enzymatic hydrolysis, sonicate for 5min every 25min, with a sonication power of 150W). Then inactivate the enzyme by incubating at 82℃ for 6min, and centrifuge at 5000rpm to obtain the supernatant.

[0056] S4. After filtering the supernatant once through a 30kDa filter membrane, collect the filtrate and then filter it a second time through a 10kDa filter membrane to obtain the silkworm polypeptide extract.

[0057] Antioxidant capacity test: The antioxidant capacity of the silkworm peptide extracts in Examples 1-6 was tested using the DPPH method. The procedure included: DPPH (1,1-diphenyl-2-trinitrophenylhydrazine) was dissolved in ethanol to prepare a 0.2 mmol / L stock solution. 1 mL of the stock solution was mixed with 1 mL of ethanol to obtain a 1,1-diphenyl-2-trinitrophenylhydrazine ethanol solution. The absorbance at 517 nm was measured and recorded as A0.

[0058] The silkworm polypeptide extracts obtained in Examples 1-6 were dissolved in ethanol to obtain the test solution. The absorbance of the test solution at 517 nm was measured and denoted as A. b .

[0059] The above test solution was mixed with an equal volume of 1,1-diphenyl-2-trinitrophenylhydrazine ethanol solution to obtain a mixed solution. The absorbance at 517 nm was measured and denoted as A. t .

[0060] .

[0061] The DPPH removal rate results are shown in Table 1 and Figure 1 .

[0062] Table 1 DPPH removal rate results

[0063] Table 1 shows that the extraction process for silkworm pupae polypeptides in this invention can efficiently retain the antioxidant activity of polypeptides. The DPPH scavenging rate of the silkworm pupae polypeptide extracts obtained in all examples reached over 80%. This effectively proves that the extraction process provided by this invention can avoid the destruction of the polypeptide spatial structure, overcome the technical defects of large polypeptide activity loss in traditional extraction processes, and the obtained polypeptide extracts have excellent antioxidant capacity, meeting the activity requirements of the core functional ingredients in skin care products.

[0064] Depend on Figure 1 The results show that the extracted silkworm polypeptide extract of the present invention exhibits excellent DPPH free radical scavenging ability, and the scavenging rates of Examples 1-6 are similar, which confirms that the silkworm polypeptide extraction process provided by the present invention has good process stability and is suitable for the application requirements of large-scale industrial production.

[0065] Application Example 1 A repair mask composition, based on 100% by weight, comprises the following raw materials: The extract prepared in Example 1 contained 3% silkworm polypeptide extract, 0.1% dipotassium glycyrrhizate, 0.2% 4-tert-butylcyclohexanol, 8% glycerol, 5% butylene glycol, 2% 1,3-propanediol, 0.1% sodium hyaluronate, 0.4% p-hydroxyacetophenone, 0.2% ethylhexylglycerin, and the remainder was water.

[0066] Application Example 2 A repair mask composition, based on 100% by weight, comprises the following raw materials: Example 2 prepared a silkworm polypeptide extract containing 6%, dipotassium glycyrrhizate 0.5%, 4-tert-butylcyclohexanol 0.2%, glycerol 8%, butylene glycol 5%, 1,3-propanediol 5%, sodium hyaluronate 0.1%, p-hydroxyacetophenone 0.2%, ethylhexylglycerin 0.1%, and the remainder being water.

[0067] Application Example 3 A repair mask composition, based on 100% by weight, comprises the following raw materials: Example 3 prepared a silkworm polypeptide extract containing 4%, dipotassium glycyrrhizate 0.2%, 4-tert-butylcyclohexanol 0.06%, glycerol 4%, butylene glycol 3%, 1,3-propanediol 3%, sodium hyaluronate 0.2%, p-hydroxyacetophenone 0.3%, ethylhexylglycerin 0.1%, and the remainder being water.

[0068] Application Example 4 A repair mask composition, based on 100% by weight, comprises the following raw materials: Example 4 prepared a silkworm polypeptide extract containing 5%, dipotassium glycyrrhizate 0.4%, 4-tert-butylcyclohexanol 0.08%, glycerol 7%, butylene glycol 4%, 1,3-propanediol 4%, sodium hyaluronate 0.2%, p-hydroxyacetophenone 0.3%, ethylhexylglycerin 0.2%, and the remainder being water.

[0069] Application Example 5 A repair mask composition, based on 100% by weight, comprises the following raw materials: Example 5 prepared a silkworm polypeptide extract containing 5%, dipotassium glycyrrhizate 0.4%, 4-tert-butylcyclohexanol 0.07%, glycerol 5%, butylene glycol 4%, 1,3-propanediol 3%, sodium hyaluronate 0.3%, p-hydroxyacetophenone 0.4%, ethylhexylglycerin 0.2%, with the remainder being water.

[0070] Application Example 6 A repair mask composition, based on 100% by weight, comprises the following raw materials: Example 6 prepared a silkworm polypeptide extract containing 5%, dipotassium glycyrrhizate 0.4%, 4-tert-butylcyclohexanol 0.08%, glycerol 6%, butylene glycol 4%, 1,3-propanediol 4%, sodium hyaluronate 0.3%, p-hydroxyacetophenone 0.4%, ethylhexylglycerin 0.2%, and the remainder being water.

[0071] Test example: 154 patients aged 25-50 years with oily skin and redness or itching were randomly selected, including 77 men and 77 women. The patients were randomly divided into 7 groups of 22 each, further subdivided into 6 experimental groups and 1 control group. In the 6 experimental groups, patients applied the repair mask compositions described in Examples 1-6, respectively, evenly to both cheeks after cleansing in the morning and evening. Patients in the control group did not perform any treatment after cleansing in the morning and evening. All 7 groups used the mask for 28 days, recording the time it took for redness / itch to subside and taking the average. After 28 days, on the 29th day, the oil content of each group's skin surface was measured using a skin oil analyzer after cleansing in the morning, and the average was taken as the initial oil content. The skin moisture content was also measured using a skin moisture analyzer, and the average was taken as the initial moisture content. Eight hours after using the repair mask composition on the morning of the 29th day, the oil and moisture content of each group's skin surface were measured again, and the average was taken as the final oil and moisture content.

[0072] The rate of change in skin surface oil content before and after use was calculated, and the results are shown in Table 2.

[0073] .

[0074] Table 2 Test Case Results

[0075] As shown in Table 2, the silkworm peptides extracted in this invention, when applied to the facial mask composition, exhibit significant anti-inflammatory, redness-reducing, moisturizing, and water-oil balance regulating effects. Redness / itching symptoms may subside in a short period, and the final skin moisture content is significantly increased compared to the initial value. The skin surface oil content is also significantly regulated, demonstrating that this facial mask composition is suitable for the repair needs of oily and sensitive skin, solving the problems of redness, dryness, itching, dehydration, and oil imbalance in this type of skin.

[0076] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A process for extracting polypeptides from Bombyx Batryticatus, characterized in that, Includes the following steps: S1. The silkworm powder was degreased by supercritical CO2 extraction to obtain degreased silkworm powder; S2. Mix defatted silkworm powder, eutectic solvent, and antioxidant, and then perform cell wall disruption treatment to obtain a cell wall disruption suspension. S3. Add the complex enzyme to the cell wall disruption suspension and perform enzymatic hydrolysis, enzyme inactivation, and centrifugation sequentially to obtain the supernatant; the complex enzyme includes papain, bromelain, and neutral protease. S4. After filtering the supernatant once through a 30kDa filter membrane, collect the filtrate and then filter it a second time through a 10kDa filter membrane to obtain the silkworm polypeptide extract.

2. The extraction process of claim 1, wherein, The supercritical CO2 extraction described in S1 is performed at a temperature of 30-40℃, a pressure of 20-25MPa, a time of 1.5-2h, and a CO2 flow rate of 20-25L / h. The mesh size of silkworm powder is 200-300 mesh.

3. The extraction process of claim 1, wherein the process is characterized by, The mass ratio of defatted silkworm powder, eutectic solvent, and antioxidant in S2 is 1:10-15:0.05-0.

1.

4. The extraction process for silkworm pupae polypeptides according to claim 1 or 3, characterized in that, The eutectic solvent in S2 is composed of choline chloride and glycerol, wherein the molar ratio of choline chloride to glycerol is 1:1.5-2; The antioxidant is vitamin C.

5. The extraction process for rich in silkworm pupae polypeptide according to claim 1, characterized in that, The cell wall breaking process described in S2 has a power of 80-120W, a time of 3-5 minutes, and a temperature of 25-35℃.

6. The extraction process for rich in silkworm pupae polypeptide according to claim 1, characterized in that, The mass ratio of the compound enzyme to defatted silkworm powder in S3 is 2-5:100; The mass ratio of papain, bromelain, and neutral protease is 1-2:1:

1.

7. The extraction process for rich in silkworm pupae polypeptide according to claim 1, characterized in that, The enzymatic hydrolysis described in S3 is carried out at a temperature of 45-50℃, a time of 3-4 hours, and a pH of 6-7. The enzymatic hydrolysis process is also accompanied by intermittent ultrasonic treatment. The intermittent ultrasonic treatment method is as follows: after enzymatic hydrolysis for 25-30 minutes, ultrasonic treatment is turned on for 3-5 minutes at a power of 150-200W. The enzyme inactivation temperature is 80-85℃ and the time is 4-6 minutes.

8. A silkworm polypeptide extract obtained by the extraction process rich in silkworm polypeptides according to any one of claims 1-7.

9. The application of the silkworm polypeptide extract according to claim 8 in a repair mask.

10. A repairing facial mask composition, characterized in that, Based on 100% by weight, the repair mask composition comprises 3%-6% of the silkworm polypeptide extract as described in claim 8, 0.1%-0.5% of dipotassium glycyrrhizate, 0.05%-0.2% of 4-tert-butylcyclohexanol, 3%-8% of glycerin, 2%-5% of butylene glycol, 2%-5% of 1,3-propanediol, 0.1%-0.3% of sodium hyaluronate, 0.2%-0.4% of p-hydroxyacetophenone, 0.1%-0.2% of ethylhexylglycerin, and the remainder is water.

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