A firming mask based on active protein of danaida cyanoxantha and its preparation process

Abstract

The present application relates to the technical field of daily cosmetics, in particular to a firming mask based on active protein of Bombyx Batryticatus and a preparation process thereof, which comprises a mask powder and a mask essence liquid independently packaged, the mask powder comprises microcapsules, the microcapsules sequentially comprise a calcium alginate gel core, an active protein layer of Bombyx Batryticatus and a chitosan outer coating film from inside to outside, trehalose and plant lactobacillus in a dormant state are embedded in the calcium alginate gel core, the active protein layer of Bombyx Batryticatus is adsorbed on the surface of the calcium alginate gel core, and the chitosan outer coating film covers the outside of the active protein layer of Bombyx Batryticatus; the mask essence liquid comprises plant lactobacillus fermentation filtrate, a moisturizing base plant extract composition and a mild preservative, and the rest is deionized water. The present application adopts powder-liquid separation independent packaging and microcapsules, plant lactobacillus and Bombyx Batryticatus are completely isolated and do not interfere with each other in the storage stage, are mixed on site to trigger step-by-step release when used, and have the advantages of long-acting stability, high skin permeability, low irritation and multi-effect synergy.

Description

Technical Field

[0001] This invention relates to the field of daily cosmetics technology, and in particular to a firming facial mask based on silkworm active protein and its preparation process. Background Technology

[0002] Silkworm extract is rich in active substances such as peptides and amino acids, and has skin care effects such as reducing fine lines, firming the skin, and soothing pigmentation. It is widely used in spot-fading and anti-aging mask products. The existing silkworm-containing masks have the following problems: (1) Poor stability of silkworm active protein: Natural silkworm protein is a large molecular weight polypeptide. It is easily denatured and inactivated by temperature, pH and light in conventional aqueous formulas. The efficacy of the product continues to decline during the shelf life and the effect of use is unstable; (2) Low transdermal absorption efficiency: The molecular weight of native silkworm protein is large and it is difficult to penetrate the stratum corneum barrier. Most of the active ingredients only stay on the skin surface and the effects of stimulating collagen, firming and reducing wrinkles in the dermis are weak; (3) Single efficacy and no synergistic system: The existing products simply combine silkworm with basic moisturizing ingredients without building a fermentation-promoting penetration and multi-plant extract synergistic system, which cannot simultaneously achieve multiple effects such as anti-wrinkle, brightening and barrier repair; (4) High irritation of the formula: Traditional masks rely on high doses of chemical preservatives to inhibit microorganisms. Sensitive skin is prone to redness and stinging. In order to make up for the poor absorption, the amount of silkworm added can only be increased, which further increases the risk of irritation. Summary of the Invention

[0003] The purpose of this invention is to provide a firming facial mask based on the active protein of silkworm and its preparation process. The mask uses powder and liquid separation and independent packaging with microcapsules. During the storage stage, Lactobacillus plantarum and silkworm protein are completely isolated and do not interfere with each other. When used, they are mixed on-site to trigger step-by-step release. It has the advantages of long-lasting stability, high transdermal permeability, low irritation and multi-effect synergy.

[0004] To achieve the above objectives, this invention provides a firming facial mask based on silkworm active protein, comprising separately packaged mask powder and mask essence. The mask powder includes microcapsules, which, from the inside out, consist of a calcium alginate gel core, a silkworm active protein interlayer, and a chitosan outer coating. The calcium alginate gel core contains trehalose and dormant Lactobacillus plantarum. The silkworm active protein interlayer is adsorbed onto the surface of the calcium alginate gel core. The chitosan outer coating covers the silkworm active protein interlayer. By mass percentage, the mask essence comprises 1%~15% Lactobacillus plantarum fermentation filtrate, 5%~15% moisturizing matrix, 2%~10% plant extract composition, 0.5%~1.5% mild preservative, and the remainder is deionized water.

[0005] Preferably, the mask powder and mask essence are individually sterile packaged. When using, the mask powder and mask essence are mixed to form a mask essence paste, which is then applied directly or used to soak the mask sheet.

[0006] Preferably, the microcapsules in the mask powder account for more than 95% of the total mass of the mask powder, and the remainder is an anti-caking agent; the ratio of mask powder to mask essence is 1~5g: 25~30g.

[0007] Preferably, the microcapsules have a particle size of 50~200μm; the active protein of *Bombyx mori* is a small molecule active peptide with a molecular weight of 3~10kDa.

[0008] Preferably, the moisturizing matrix includes one or more of glycerin, butylene glycol, sodium hyaluronate, and panthenol.

[0009] Preferably, the plant extract composition includes multiple extracts of Tremella fuciformis extract, mulberry leaf extract, safflower extract, ginseng root extract, Poria cocos extract, Ampelopsis japonica extract, and Atractylodes macrocephala extract.

[0010] Preferably, the mild preservative is composed of p-hydroxyacetophenone, 1,2-hexanediol and octyl glycol in a mass ratio of 4:6:1.

[0011] This invention also provides a preparation process for the above-mentioned firming facial mask based on silkworm active protein, comprising the following steps: S1. Preparation of concentrated small molecule active peptides from silkworm: After defatting the silkworm raw material, it was enzymatically hydrolyzed at low temperature (45-55℃) for 2-3 hours. After inactivation and centrifugation, it was subjected to fractional ultrafiltration using 3kDa and 10kDa molecular weight cutoff membranes to collect the 3-10kDa fraction and obtain concentrated small molecule active peptides from silkworm. S2. Preparation of calcium alginate gel core: Dormant Lactobacillus plantarum, sodium alginate aqueous solution and trehalose are mixed evenly to obtain a mixture. The mixture is then dripped into a calcium ion crosslinking solution and solidified to obtain calcium alginate gel microspheres with dormant Lactobacillus plantarum and trehalose embedded inside. S3. Microcapsule preparation: The calcium alginate gel microspheres of S2 are immersed in the concentrated solution of small molecule active peptides of silkworm pupae from S1, and stirred for adsorption, so that a uniform layer of active protein from silkworm pupae is formed on the outer wall of the calcium alginate gel microspheres; then immersed in chitosan film-forming solution, and solidified by emulsification-internal gelation method to form a chitosan outer coating membrane, and dried to obtain microcapsules; the microcapsules are mixed with an anti-caking agent to obtain a face mask powder; S4. Preparation of fermentation filtrate: Take the Lactobacillus plantarum strain, inoculate the basic culture medium and anaerobic ferment for 24 hours, then add the mixed extract of Tremella fuciformis and mulberry leaves and microaerobic ferment for 18 hours, centrifuge, and filter sterilize through a 0.22μm filter membrane to obtain Lactobacillus plantarum fermentation filtrate rich in organic acids and hydrolytic enzymes. S5. Preparation of essence: Under normal temperature conditions, the fermentation filtrate of Lactobacillus plantarum in S4, moisturizing matrix, plant extract composition, mild preservative and deionized water are added to the mixing tank in sequence and mixed evenly. The pH of the system is adjusted to 5.5~6.0. After high pressure homogenization at 30~800 bar for 2~5 minutes, the mixture is filtered and aged in the dark for 12 hours to obtain the mask essence. S6. Finished Product Packaging: The mask powder obtained in S3 and the mask essence obtained in S5 are aseptically and independently packaged and combined to finally produce a firming mask based on silkworm active protein.

[0012] Preferably, in S2, the mass ratio of dormant Lactobacillus plantarum, sodium alginate aqueous solution, and trehalose is 4~6:84~91:5~10.

[0013] Preferably, in S3, the chitosan film-forming solution is an aqueous solution of chitosan, gelatin, and glacial acetic acid, with a chitosan mass fraction of 0.2%~0.5%, a gelatin mass fraction of 0.1%~0.3%, a glacial acetic acid mass fraction of 0.05%~0.1%, and the remainder being sterile water.

[0014] Mechanism of the invention: When used, the microcapsules are mixed with a mask essence with a pH of 5.5~6.0, conforming to the skin's slightly acidic environment, triggering a layered and orderly release: 1) Slight acid penetration dissolves the outer layer of chitosan: In the slightly acidic environment of the skin, chitosan amino groups are protonated, causing the outer layer of chitosan to swell and rupture, preferentially releasing 3~10kDa silkworm small molecule active peptides from the interlayer, which directly contact the stratum corneum; 2) Slight acid continuously penetrates the calcium alginate gel core: After the outer layer of chitosan is damaged, water and slight acid enter the calcium alginate gel core, the trehalose glass matrix dissolves upon contact with water, and the plant lactobacillus is released from dormancy and quickly resumes metabolism; 3) Plant lactobacillus generates penetration-promoting organic acids in situ: The revived plant lactobacillus continuously secretes lactic acid, short-chain fatty acids, and hydrolytic enzymes, softening the intercellular spaces of the stratum corneum, opening the skin's absorption channels, and simultaneously degrading the stratum corneum barrier, significantly increasing the dermal penetration of silkworm active peptides. Meanwhile, the mask essence contains pre-prepared Lactobacillus plantarum fermentation filtrate, rich in organic acids and hydrolytic enzymes. After mixing, it further helps soften the stratum corneum and forms a dual penetration-enhancing system with the organic acids released by the microcapsules. The moisturizing matrix constructs hydration channels to maintain the stratum corneum's moisture and provides a water environment for the penetration of active ingredients, while repairing the dry and damaged skin barrier. The plant extract composition works synergistically to achieve firming, spot reduction, brightening, and repair. The gentle preservative system reduces the addition of irritating preservatives, making it suitable for sensitive skin.

[0015] The beneficial effects of this invention are: (1) This invention uses separate packaging for powder and liquid, with the active protein of silkworm pupae embedded in microcapsules, completely isolated from the aqueous phase of the mask essence. This fundamentally avoids the defect of active protein in traditional aqueous formulations being deactivated due to hydrolysis and oxidation. It can be reconstituted immediately upon use and activated immediately upon application, ensuring that it is in a highly active state every time it is used, thus completely solving the industry pain point of continuous decline in efficacy during the shelf life of conventional masks.

[0016] (2) This invention achieves precise control of the timing of first promoting penetration, then tightening, and finally repairing by combining microcapsules with the stratum corneum softening effect of organic acids in the fermentation filtrate of Lactobacillus plantarum. The small molecule active peptides of Bombyx mori are concentrated and penetrated during the optimal window period when the stratum corneum barrier is softened, effectively reaching the dermis to stimulate collagen regeneration. The organic acids and bacteriocins produced by Lactobacillus plantarum during the application of the mask continue to act on the stratum corneum. The probiotic metabolites remaining after the mask is removed can still play a role in microecological regulation, combining immediate nourishment and long-term repair, and greatly improving the bioavailability of active ingredients.

[0017] (3) This invention relies on natural metabolites such as lactic acid and bacteriocins in the fermentation filtrate to inhibit harmful microorganisms, significantly reducing the amount of chemical preservatives used, greatly improving the gentleness of the formula, and making it more suitable for sensitive skin. At the same time, the three pathways of firming and anti-wrinkle of silkworm small molecule active peptides, brightening and fading of spots by fermentation filtrate, and moisturizing and repairing by multi-plant extract composition work synergistically, so that a single product can simultaneously achieve multiple effects such as firming and lifting, fading fine lines, brightening skin tone, and repairing the skin barrier, meeting consumers' needs for efficient and simple skin care.

[0018] 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

[0019] Figure 1 This is a schematic diagram showing the results of a 4-week efficacy trial on humans for this invention. Detailed Implementation

[0020] This invention provides a firming facial mask based on silkworm active protein, comprising separately packaged mask powder and mask essence. The mask powder includes microcapsules, which, from the inside out, consist of a calcium alginate gel core, a silkworm active protein interlayer, and a chitosan outer coating. The calcium alginate gel core contains trehalose and dormant Lactobacillus plantarum. The silkworm active protein interlayer is adsorbed onto the surface of the calcium alginate gel core. The chitosan outer coating covers the silkworm active protein interlayer. By mass percentage, the mask essence comprises 1%~15% Lactobacillus plantarum fermentation filtrate, 5%~15% moisturizing matrix, 2%~10% plant extract composition, 0.5%~1.5% mild preservative, and the remainder is deionized water.

[0021] The microcapsule structure in this invention ensures that *Lactobacillus plantarum* and *Bombyx mori* active proteins remain completely isolated during storage, preventing mutual degradation reactions. The trehalose core forms a glassy matrix that replaces water molecules to encapsulate the *Lactobacillus plantarum* cell membrane, locking in the bacterial structure and inhibiting bacterial metabolism and acid production. Throughout storage, the probiotics remain in a dormant, seemingly dead state, preventing premature production of organic acids that could damage the *Bombyx mori* active peptides. The outer chitosan coating blocks light, temperature, trace amounts of moisture, and oxygen, preventing oxidation and spatial structural denaturation of the small-molecule peptides from *Bombyx mori*. Furthermore, the microcapsule mask powder remains completely isolated from the aqueous essence, preventing premature swelling and failure of the chitosan coating due to prolonged water immersion, thus extending the shelf life of the active ingredients from the source.

[0022] Preferably, the mask powder and mask essence are individually aseptically packaged. When using, the mask powder and mask essence are mixed to form a mask essence paste, which is then applied directly or used to soak a mask sheet. This invention, through its two-component independent packaging, solves the industry problem of premature revival of *Lactobacillus plantarum* and denaturation and inactivation of *Bombyx mori* proteins at the root of storage, significantly extending the product's shelf life.

[0023] Preferably, the microcapsules in the mask powder account for more than 95% of the total mass of the mask powder, with the remainder being an anti-caking agent; the ratio of mask powder to mask essence is 1~5g: 25~30g. A small amount of anti-caking agent prevents the microcapsule powder from clumping, and the ratio is suitable for a single facial application, balancing active ingredient concentration and skin feel. The anti-caking agent can be corn starch or talc, commonly used in the prior art; this invention does not impose further limitations.

[0024] Preferably, the microcapsules have a particle size of 50~200μm; the active protein of *Bombyx mori* is a small molecule active peptide with a molecular weight of 3~10kDa. Microcapsules of this particle size are evenly dispersed after mixing, without any particulate or foreign body sensation; the 3~10kDa small molecule peptide can quickly penetrate the stratum corneum without secondary degradation, improving skin absorption efficiency.

[0025] Preferably, the moisturizing matrix includes one or more of glycerin, butylene glycol, sodium hyaluronate, and panthenol. The polyols, combined with sodium hyaluronate and panthenol, form a multi-layered water-locking system, improving dry and rough skin while providing a hydrating medium for the penetration of active ingredients.

[0026] Preferably, the plant extract composition includes multiple extracts of Tremella fuciformis extract, mulberry leaf extract, safflower extract, ginseng root extract, Poria cocos extract, Ampelopsis japonica extract, and Atractylodes macrocephala extract.

[0027] In some specific embodiments of the present invention, the plant extract composition simultaneously comprises Tremella fuciformis extract, mulberry leaf extract, safflower extract, ginseng root extract, Poria cocos extract, Ampelopsis japonica extract, and Atractylodes macrocephala extract. The plant extract composition of the present invention forms a complete repairing, brightening, and antioxidant complex system. Tremella fuciformis polysaccharides repair the damaged skin barrier, mulberry leaf and safflower inhibit tyrosinase to lighten pigmentation, and ginseng and Atractylodes macrocephala promote collagen regeneration and tighten loose skin.

[0028] Preferably, the mild preservative is composed of p-hydroxyacetophenone, 1,2-hexanediol, and octyl glycol in a mass ratio of 4:6:1. The mild preservative of this invention relies on polyols for antibacterial activity, significantly reducing the addition of traditional irritating preservative ingredients, making it safe for sensitive skin.

[0029] This invention also provides a preparation process for the above-mentioned firming facial mask based on silkworm active protein, comprising the following steps: S1. Preparation of concentrated small molecule active peptides from silkworm pupae: After defatting the silkworm pupae raw material, low-temperature enzymatic hydrolysis was performed using a compound enzyme consisting of alkaline protease and flavor protease in a mass ratio of 1:1. The hydrolysis temperature was 45-55℃ and the hydrolysis time was 2-3 hours. After enzyme inactivation and centrifugation, the mixture was subjected to fractional ultrafiltration using 3kDa and 10kDa molecular weight cutoff filter membranes. The 3-10kDa fraction was collected to obtain concentrated small molecule active peptides from silkworm pupae. S2. Preparation of calcium alginate gel core: Dormant Lactobacillus plantarum, sodium alginate aqueous solution and trehalose are mixed evenly to obtain a mixture. The mixture is then dripped into a calcium ion crosslinking solution and solidified to obtain calcium alginate gel microspheres with dormant Lactobacillus plantarum and trehalose embedded inside. S3. Microcapsule preparation: The calcium alginate gel microspheres of S2 are immersed in the concentrated solution of small molecule active peptides of silkworm pupae from S1, and stirred for adsorption, so that a uniform layer of active protein from silkworm pupae is formed on the outer wall of the calcium alginate gel microspheres; then immersed in chitosan film-forming solution, and solidified by emulsification-internal gelation method to form a chitosan outer coating membrane, and dried to obtain microcapsules; the microcapsules are mixed with an anti-caking agent to obtain a face mask powder; S4. Preparation of fermentation filtrate: Take the Lactobacillus plantarum strain, inoculate the basic culture medium and anaerobic ferment for 24 hours, then add the mixed extract of Tremella fuciformis and mulberry leaves and microaerobic ferment for 18 hours, centrifuge, and filter sterilize through a 0.22μm filter membrane to obtain Lactobacillus plantarum fermentation filtrate rich in organic acids and hydrolytic enzymes. S5. Preparation of essence: Under normal temperature conditions, the fermentation filtrate of Lactobacillus plantarum in S4, moisturizing matrix, plant extract composition, mild preservative and deionized water are added to the mixing tank in sequence and mixed evenly. The pH of the system is adjusted to 5.5~6.0. After high pressure homogenization at 30~800 bar for 2~5 minutes, the mixture is filtered and aged in the dark for 12 hours to obtain the mask essence. S6. Finished Product Packaging: The mask powder obtained in S3 and the mask essence obtained in S5 are aseptically and independently packaged and combined to finally produce a firming mask based on silkworm active protein.

[0030] Preferably, in S2, the mass ratio of dormant *Lactobacillus plantarum*, sodium alginate aqueous solution, and trehalose is 4~6:84~91:5~10. This ratio ensures that trehalose adequately coats the bacterial cells to achieve dormancy, while the sodium alginate ensures complete cross-linking and formation of the microspheres, making them less prone to breakage.

[0031] Preferably, in step S3, the chitosan film-forming solution is an aqueous solution composed of chitosan, gelatin, and glacial acetic acid, with a chitosan mass fraction of 0.2%~0.5%, a gelatin mass fraction of 0.1%~0.3%, and a glacial acetic acid mass fraction of 0.05%~0.1%, the remainder being sterile water. Gelatin enhances the toughness of the film layer, while glacial acetic acid regulates the dissolution environment of chitosan, resulting in a dense film with pH-responsive dissolution characteristics.

[0032] The emulsification-internal gelation method in S3 of this invention can be achieved using existing technology, and no further limitations are imposed.

[0033] The preparation process of this invention adopts mild low-temperature treatment throughout, without using high temperature to destroy the protein, Lactobacillus plantarum and plant extract activity. The microcapsule layer preparation realizes the partitioning and isolation of active ingredients. The two-component independent packaging process is simple and adaptable to conventional cosmetic aseptic filling production lines. Mass production can be carried out without the need to modify large equipment.

[0034] 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.

[0035] Example 1 This invention provides a firming facial mask based on active proteins from silkworm pupae, comprising individually packaged mask powder and mask essence. The mask powder includes microcapsules, and the mask essence includes Lactobacillus plantarum fermentation filtrate, a moisturizing matrix, a composition of plant extracts, a mild preservative, and deionized water.

[0036] The above-mentioned process for preparing a firming facial mask based on the active protein of silkworm pupae includes the following steps: S1, preparation of concentrated silkworm pupae small molecule active peptide solution: 1 part of dried silkworm pupae is pulverized and defatted, and then prepared into a 10% (w / w) silkworm pupae powder aqueous suspension. The pH is adjusted to 8.0, and low-temperature enzymatic hydrolysis is performed using a compound enzyme of alkaline protease and flavor protease in a mass ratio of 1:1. The total amount of compound enzyme added is 0.3% of the suspension mass, the enzymatic hydrolysis temperature is 50℃, and the enzymatic hydrolysis time is 2.5h. After centrifugation with enzyme inactivation at 90℃, the solution is subjected to fractional ultrafiltration using 3kDa and 10kDa molecular weight cutoff filter membranes, and the 3~10kDa fraction is collected to obtain the concentrated silkworm pupae small molecule active peptide solution.

[0037] S2. Preparation of calcium alginate gel core: 5 parts of dormant Lactobacillus plantarum bacterial solution, 87 parts of sodium alginate aqueous solution and 7 parts of trehalose are mixed evenly to obtain a mixed solution. The mixed solution is then dripped into a calcium chloride crosslinking solution to solidify and form calcium alginate gel microspheres with dormant Lactobacillus plantarum and trehalose embedded inside.

[0038] S3. Microcapsule preparation: The calcium alginate gel microspheres of S2 were immersed in the concentrated solution of small molecule active peptides of silkworm pupae from S1, and stirred for adsorption, so that a uniform layer of active silkworm pupae protein was formed on the outer wall of the calcium alginate gel microspheres; then immersed in a chitosan film-forming solution (0.3% chitosan, 0.2% gelatin, 0.08% glacial acetic acid, and the remainder being sterile water), and solidified by the emulsification-internal gelation method to form a chitosan outer coating film. After drying, microcapsules with a particle size of 50~200μm were obtained; 98 parts of microcapsules were mixed with 2 parts of corn starch to prepare the face mask powder.

[0039] S4. Preparation of fermentation filtrate: Take the Lactobacillus plantarum strain, inoculate the basic culture medium and anaerobic ferment for 24 hours, then add the mixed extract of Tremella fuciformis and mulberry leaves and microaerobic ferment for 18 hours, centrifuge, and filter sterilize through a 0.22μm filter membrane to obtain Lactobacillus plantarum fermentation filtrate rich in organic acids and hydrolytic enzymes. S5. Essence Preparation: Under normal temperature conditions, add the following ingredients by weight percentage: 8% S4 fermentation filtrate, 3.5% glycerin, 2% butylene glycol, 0.15% sodium hyaluronate, 0.3% panthenol, 3% tremella extract, 2% mulberry leaf extract, 1.5% safflower extract, 0.8% ginseng root extract, 0.6% poria cocos extract, 0.5% white peony root extract, 0.5% atractylodes macrocephala extract, 0.4% p-hydroxyacetophenone, 0.6% 1,2-hexanediol, and 0.1% caprylyl glycol. Add deionized water to bring the total to 100%. Mix thoroughly in a stirring vessel, adjust the pH of the system to 5.8, homogenize under 500 bar for 5 minutes, filter, and mature in the dark for 12 hours to obtain the mask essence.

[0040] S6. Finished Product Packaging: The mask powder obtained in S3 and the mask essence obtained in S5 are aseptically and individually packaged and combined to finally produce a firming mask based on silkworm active proteins. To use, add 2g of powder to 28g of essence and stir to form an essence paste. Apply to the face and leave for 25 minutes.

[0041] Example 2 The difference from Example 1 is that in S2, 4 parts of dormant Lactobacillus plantarum, 84 parts of sodium alginate aqueous solution, and 5 parts of trehalose are mixed evenly to obtain a mixed solution, and the mass percentage of the fermentation filtrate in S5 is 1%, while the rest are the same as in Example 1.

[0042] Example 3 The difference from Example 1 is that in S2, 6 parts of dormant Lactobacillus plantarum, 91 parts of sodium alginate aqueous solution, and 10 parts of trehalose are mixed evenly to obtain a mixed solution, and the mass percentage of the fermentation filtrate in S5 is 15%, and the rest are the same as in Example 1.

[0043] Comparative Example 1 A facial mask essence comprises silkworm small molecule active peptides, dormant Lactobacillus plantarum bacterial solution, Lactobacillus plantarum fermentation filtrate, moisturizing matrix, plant extract composition, mild preservative and deionized water.

[0044] The preparation process includes the following steps: Under normal temperature conditions, 8% by mass of *Lactobacillus plantarum* fermentation filtrate, 3.5% glycerol, 2% butylene glycol, 0.15% sodium hyaluronate, 0.3% panthenol, 3% *Tremella fuciformis* extract, 2% mulberry leaf extract, 1.5% safflower extract, 0.8% ginseng root extract, 0.6% *Poria cocos* extract, 0.5% *Ampelopsis japonica* extract, 0.5% *Atractylodes macrocephala* extract, 0.4% p-hydroxyacetophenone, 0.6% 1,2-hexanediol, 0.1% caprylyl glycol, 5% dormant *Lactobacillus plantarum* bacterial culture, and 5% *Bombyx mori* small molecule active peptide concentrate are added to a stirring vessel and mixed evenly. The pH of the system is adjusted to 5.8, and the mixture is homogenized under high pressure at 500 bar for 5 minutes. After filtration, the mixture is aged in the dark for 12 hours to obtain the mask essence. The preparation of the *Bombyx mori* small molecule active peptide concentrate and the fermentation filtrate are the same as in Example 1.

[0045] Comparative Example 2 The difference from Example 1 is that S4 was not performed. The composition of the mask essence by weight percentage is as follows: 3.5% glycerin, 2% butylene glycol, 0.15% sodium hyaluronate, 0.3% panthenol, 3% tremella fuciformis extract, 2% mulberry leaf extract, 1.5% safflower extract, 0.8% ginseng root extract, 0.6% poria cocos extract, 0.5% ampelopsis japonica extract, 0.5% atractylodes macrocephala extract, 0.4% p-hydroxyacetophenone, 0.6% 1,2-hexanediol, 0.1% caprylyl glycol, and the balance being deionized water. All other components are the same as in Example 1.

[0046] Comparative Example 3 The difference from Example 1 is that S1 and S3 were not performed; only the calcium alginate gel core was prepared. The rest is the same as in Example 1.

[0047] Performance testing The facial masks prepared in Examples 1-3 and Comparative Examples 1-3 were subjected to four parallel tests according to a unified testing standard. Each group of samples was tested three times in parallel, and the average value was taken. The test methods are as follows: (1) Stability test of silkworm protein: The sample was stored in a 25℃, 500lx light-proof constant temperature incubator for 24 months, and the proportion of residual active protein to the initial content was detected by BCA protein quantification method. (2) In vitro transdermal absorption rate: Franz diffusion cell of isolated porcine skin was used. The receiving liquid was sampled after 24 hours. The amount of skeletal peptides permeated was determined by high performance liquid chromatography, and the transdermal absorption rate was calculated. (3) HET-CAM eye irritation test: The chorioallantoic membrane of chicken embryos incubated for 10-12 days was used. The NC stimulation value was calculated according to the standard. NC < 1 means no irritation, 1-3 means mild irritation, and 3-5 means moderate irritation. (4) Human efficacy trial over 4 weeks: 90 healthy volunteers aged 30-45 years with facial sagging, fine lines, and dullness were selected and divided into 6 groups. The treatment was administered 3 times per week for 4 consecutive weeks. The R2 elasticity improvement rate was measured using a skin elasticity meter, and the facial melanin reduction rate was measured using a spectrophotometer. The results are shown in Table 1 and... Figure 1 .

[0048] Table 1. Test data of the face masks prepared in Examples 1-3 and Comparative Examples 1-3

[0049] As shown in Table 1, after 24 months of storage, the residual activity of the silkworm protein in Examples 1-3 of this invention was all above 83%. In Comparative Example 1, without microcapsules, the active ingredient was directly miscible with the aqueous phase, resulting in significant protein oxidative degradation, leaving only 20.3% activity. In Comparative Example 3, without a microcapsule layer between the silkworm protein and the aqueous phase, the protein could not be isolated from light, and the degree of protein inactivation was similar to that of Comparative Example 1. This demonstrates that the independent packaging of microcapsules and powder / liquid is the core key structure for maintaining the long-term stability of silkworm protein.

[0050] The transdermal absorption rates of Examples 1-3 all exceeded 82%; Comparative Example 2, without the addition of fermentation filtrate and lacking the keratin-softening organic acids, saw its transdermal absorption rate drop significantly to 35.8%; Comparative Examples 1 and 3 exhibited the worst transdermal effects. This indicates that the *Lactobacillus plantarum* fermentation filtrate and the in-situ acid production from the microcapsules synergistically enhance permeation, significantly improving the permeation efficiency of *Bombyx mori* peptides.

[0051] The NC stimulation values ​​of Examples 1-3 of the present invention are all below 1, which is determined to be non-irritating and suitable for sensitive skin. In Comparative Examples 1 and 3, probiotics and active proteins coexist directly, which easily disrupts the microbial balance of the system, increases the preservative load, and the NC value is close to 3, which poses a slight risk of irritation. The present invention relies on probiotic metabolites to assist in antibacterial activity, which can significantly reduce the amount of chemical preservatives added, and has obvious advantages in gentleness.

[0052] like Figure 1 As shown, the skin elasticity improvement and melanin fading indicators of Examples 1-3 of the present invention are better than those of Comparative Examples 1-3, indicating that the composite system of the present invention can simultaneously achieve the dual skin care effects of firming and anti-wrinkle, and brightening and fading spots.

[0053] 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 firming facial mask based on active proteins from silkworm pupae, characterized in that: The product includes individually packaged mask powder and mask essence. The mask powder comprises microcapsules, which, from the inside out, consist of a calcium alginate gel core, a silkworm active protein interlayer, and a chitosan outer coating. The calcium alginate gel core contains trehalose and dormant Lactobacillus plantarum. The silkworm active protein interlayer is adsorbed onto the surface of the calcium alginate gel core. The chitosan outer coating covers the silkworm active protein interlayer. By weight percentage, the mask essence comprises 1%–15% Lactobacillus plantarum fermentation filtrate, 5%–15% moisturizing matrix, 2%–10% plant extract composition, 0.5%–1.5% mild preservative, and the remainder is deionized water.

2. A firming facial mask based on silkworm active protein according to claim 1, characterized in that: The mask powder and mask essence are individually sterile packaged. When using, mix the mask powder and mask essence to form a mask essence paste, and then apply it directly or soak the mask sheet for use.

3. A firming facial mask based on silkworm active protein according to claim 1, characterized in that: Microcapsules account for more than 95% of the total mass of the mask powder, with the remainder being anti-caking agents; the ratio of mask powder to mask essence is 1~5g: 25~30g.

4. A firming facial mask based on silkworm active protein according to claim 1, characterized in that: The microcapsules have a particle size of 50~200μm; the active protein of *Bombyx mori* is a small molecule active peptide with a molecular weight of 3~10kDa.

5. A firming facial mask based on silkworm active protein according to claim 1, characterized in that: The moisturizing base includes one or more of glycerin, butylene glycol, sodium hyaluronate, and panthenol.

6. A firming facial mask based on silkworm active protein according to claim 1, characterized in that: The plant extract composition includes multiple extracts from Tremella fuciformis, mulberry leaf, safflower, ginseng root, Poria cocos, Ampelopsis japonica, and Atractylodes macrocephala.

7. A firming facial mask based on silkworm active protein according to claim 1, characterized in that: The mild preservative is composed of p-hydroxyacetophenone, 1,2-hexanediol and octyl glycol in a mass ratio of 4:6:

1.

8. A preparation process for a firming facial mask based on *Bombyx mori* active protein as described in any one of claims 1-7, characterized in that: Includes the following steps: S1. Preparation of concentrated small molecule active peptides from silkworm: After defatting the silkworm raw material, it was enzymatically hydrolyzed at low temperature (45-55℃) for 2-3 hours. After inactivation and centrifugation, it was subjected to fractional ultrafiltration using 3kDa and 10kDa molecular weight cutoff membranes to collect the 3-10kDa fraction and obtain concentrated small molecule active peptides from silkworm. S2. Preparation of calcium alginate gel core: Dormant Lactobacillus plantarum, sodium alginate aqueous solution and trehalose are mixed evenly to obtain a mixture. The mixture is then dripped into a calcium ion crosslinking solution and solidified to obtain calcium alginate gel microspheres with dormant Lactobacillus plantarum and trehalose embedded inside. S3. Microcapsule preparation: The calcium alginate gel microspheres of S2 are immersed in the concentrated solution of small molecule active peptides of silkworm pupae from S1, and stirred for adsorption, so that a uniform layer of active protein from silkworm pupae is formed on the outer wall of the calcium alginate gel microspheres; then immersed in chitosan film-forming solution, and solidified by emulsification-internal gelation method to form a chitosan outer coating membrane, and dried to obtain microcapsules; the microcapsules are mixed with an anti-caking agent to obtain a face mask powder; S4. Preparation of fermentation filtrate: Take the Lactobacillus plantarum strain, inoculate the basic culture medium and anaerobic ferment for 24 hours, then add the mixed extract of Tremella fuciformis and mulberry leaves and microaerobic ferment for 18 hours, centrifuge, and filter sterilize through a 0.22μm filter membrane to obtain Lactobacillus plantarum fermentation filtrate rich in organic acids and hydrolytic enzymes. S5. Preparation of essence: Under normal temperature conditions, the fermentation filtrate of Lactobacillus plantarum in S4, moisturizing matrix, plant extract composition, mild preservative and deionized water are added to the mixing tank in sequence and mixed evenly. The pH of the system is adjusted to 5.5~6.

0. After high pressure homogenization at 30~800 bar for 2~5 minutes, the mixture is filtered and aged in the dark for 12 hours to obtain the mask essence. S6. Finished Product Packaging: The mask powder obtained in S3 and the mask essence obtained in S5 are aseptically and independently packaged and combined to finally produce a firming mask based on silkworm active protein.

9. The preparation process of the firming mask based on silkworm active protein according to claim 8, characterized in that: In S2, the mass ratio of dormant Lactobacillus plantarum, sodium alginate aqueous solution, and trehalose is 4~6:84~91:5~10.

10. The preparation process of the firming mask based on silkworm active protein according to claim 8, characterized in that: In S3, the chitosan film-forming solution is an aqueous solution of chitosan, gelatin, and glacial acetic acid, with a chitosan mass fraction of 0.2%~0.5%, a gelatin mass fraction of 0.1%~0.3%, a glacial acetic acid mass fraction of 0.05%~0.1%, and the remainder being sterile water.

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