Fiber mask containing sodium hyaluronate and collagen and preparation method thereof

Abstract

The invention discloses a fibrous mask containing sodium hyaluronate and collagen and a preparation method of the fibrous mask, and relates to the technical field of mask preparation. The fiber mask comprises the following components in parts by weight: 2-10 parts of compound molecular weight sodium hyaluronate (low / medium / high molecular weight), 0.1-2 parts of compound molecular weight collagen (small molecule / large molecule) and 88-97.9 parts of solvent. The sodium hyaluronate is extracted by combining a fermentation method with multistage purification, and the collagen is prepared by combining a genetic engineering method with the fermentation method, so that the high activity and purity of the sodium hyaluronate and the collagen are kept; and forming the fiber mask through electrostatic spinning. Through simple component design, an efficient purification process and an electrostatic spinning technology, collaborative optimization of high safety, high activity and high fitness of the mask is realized, and the moisturizing effect and the use experience of the mask are remarkably improved.

Description

Technical Field

[0001] The present invention relates to the technical field of facial mask preparation, in particular to a fiber facial mask containing sodium hyaluronate and collagen and a preparation method thereof. Background Art

[0002] As people pay more and more attention to skin care, facial masks, as a convenient and effective skin care product, are becoming more and more popular among consumers. Fiber masks occupy a key position in the market due to their good adhesion and ability to carry essence.

[0003] Sodium hyaluronate has excellent moisturizing properties and can form a breathable film on the skin surface to lock in moisture, keeping the skin moist and soft; collagen is one of the main components of the skin and can maintain the skin's elasticity and tension, delaying skin aging.

[0004] However, there are some problems with the fiber masks containing sodium hyaluronate and collagen on the market. On the one hand, the ingredients of some masks are complex. In addition to sodium hyaluronate and collagen, a variety of preservatives, fragrances, pigments and other chemicals are added. These ingredients may cause irritation to the skin, especially for people with sensitive skin, who are prone to allergic reactions and other adverse reactions after use. On the other hand, the existing methods for extracting sodium hyaluronate and collagen and making fiber masks are relatively simple, resulting in low purity and activity of the extracted sodium hyaluronate and collagen, and the texture and performance of the fiber mask are not stable enough, which affects the use effect of the mask. For example, the traditional method of extracting sodium hyaluronate may not be able to completely remove impurities. The structure of collagen may be destroyed during the extraction process due to the simple process, and the original biological activity may be lost. In the production process of fiber masks, the simple preparation process may lead to uneven distribution of fibers, and the water absorption and water retention of the mask are poor.

[0005] Therefore, there is an urgent need for a fiber mask containing sodium hyaluronate and collagen with simple ingredients, more scientific and complex extraction and preparation methods, and a preparation method thereof, so as to improve the quality and use effect of the mask. Summary of the Invention

[0006] The object of the present invention is to provide a fiber mask containing sodium hyaluronate and collagen and a preparation method thereof, so as to solve the problems raised in the above background technology.

[0007] In order to solve the above technical problems, the present invention provides the following technical solutions: A fiber mask containing sodium hyaluronate and collagen, characterized in that it is prepared by electrostatic spinning of a spinning solution, wherein the spinning solution comprises the following components in parts by weight: 2-10 parts of sodium hyaluronate, 0.1-2 parts of collagen, and 88-97.9 parts of a solvent.

[0008] Furthermore, the sodium hyaluronate is compounded by sodium hyaluronate of different molecular weights, specifically including the following components in parts by weight: low molecular weight sodium hyaluronate: 20-30 parts; medium molecular weight sodium hyaluronate: 30-50 parts; high molecular weight sodium hyaluronate: 20-50 parts.

[0009] Furthermore, the low molecular weight sodium hyaluronate has a molecular weight of 5-10 kDa; the medium molecular weight sodium hyaluronate has a molecular weight of 200-800 kDa; and the high molecular weight sodium hyaluronate has a molecular weight of 1000-2000 kDa.

[0010] Furthermore, the collagen is compounded by collagens of different molecular weights, specifically including the following components in parts by weight: small molecule collagen: 60-80 parts; large molecule collagen: 20-40 parts.

[0011] Furthermore, the molecular weight of the small molecule collagen is 2-5 kDa; the molecular weight of the large molecule collagen is 80-100 kDa.

[0012] Furthermore, the solvent formula is: deionized water and an organic solvent are compounded in a volume ratio of 1-2:1; the organic solvent is selected from one of methyl ethyl ketone, N-methyl pyrrolidone, 1,3-propylene glycol, 1,2-butanediol, 1,4-butanediol, 2,3-butanediol, diacetone alcohol, acetone, n-butanol, and propanol.

[0013] A method for preparing a fiber mask containing sodium hyaluronate and collagen comprises the following steps: S1-1 Spinning solution preparation: Sodium hyaluronate, collagen, and solvent are added in sequence according to the formula, and stirred at 200-400 rpm for 4-6 hours to obtain a mixed solution; the mixed solution is filtered through a 500-mesh filter cloth to obtain a spinning solution; S1-2 Electrospinning: Inject the spinning solution into the syringe and connect it to the peristaltic pump and air pump; start the peristaltic pump, air pump and high-voltage power supply, and continue spinning for 60-120 minutes; after the spinning is completed, remove the fiber membrane from the receiving device, cut it according to the mask mold, and trim the edges to obtain a fiber mask.

[0014] Furthermore, the electrospinning conditions are: negative pressure 30-60 kPa, temperature 20-30° C., humidity 20-30%, spinning rate 0.2-0.4 L / h, and receiving distance 50 cm.

[0015] Furthermore, the thickness of the non-woven fabric-like fiber membrane is 0.1-0.5 mm, and the spinning solution advancing rate is 0.2-0.4 L / h.

[0016] Furthermore, the method for extracting sodium hyaluronate comprises the following steps: S2-1 Bacterial culture: Inoculate the bacteria into liquid culture medium and culture at 30-37°C, 150-200 rpm for 12-24 hours. 600 When the value reaches 0.6-0.8, the seed solution is obtained; S2-2 Fermentation culture: Inoculate the seed liquid into a stainless steel fermentation tank at an inoculum rate of 10%-20% for fermentation culture for 24-48 hours; S2-3 crude extraction: collect the fermentation broth and centrifuge it at 8000-10000 rpm for 10-15 minutes to remove the bacteria and collect the supernatant; add 2-3 volumes of anhydrous ethanol to the supernatant, stir and let it stand for 2-3 hours, centrifuge it at 8000-10000 rpm for 10-20 minutes, and collect the hyaluronic acid precipitate; remove impure cations through cationic resin, then elute with NaCl solution to replace it with the sodium salt form to produce sodium hyaluronate; S2-4 purification: add trypsin at 0.5-1wt% of the mass of sodium hyaluronate, incubate at 45-55°C for 2-4 hours to degrade residual protein impurities; add deoxyribonuclease at 0.5-1wt% of the mass of sodium hyaluronate and ribonuclease at 0.5-1wt% of the mass of sodium hyaluronate to remove bacterial nucleic acid contamination; add activated carbon at 0.5-1wt% of the mass of sodium hyaluronate, stir for 30-45 minutes, and filter to remove pigments and small molecular impurities; further remove endotoxins, inorganic salts and charged impurities through anion exchange resin; filter the purified solution through a 0.45μm sterilizing filter membrane, package and freeze-dry, and obtain sodium hyaluronate of different molecular weights by adjusting the dissolved oxygen concentration and the concentration of the culture medium during fermentation.

[0017] Furthermore, the bacterial species in S2-1 is one of Streptococcus, fermentation yeast, and acid-producing Bacillus.

[0018] Furthermore, the components of the liquid culture medium and the culture medium in the stainless steel fermenter are: 10-50 g / L glucose, 1-10 g / L yeast extract powder, 1-10 g / L peptone, 0.5-1 g / L MgSO4·7H2O, and 1-2 g / L KH2PO4.

[0019] Furthermore, the cationic resin is hexadecyltrimethylammonium bromide.

[0020] Furthermore, the anion resin is diethylaminoethyl cellulose.

[0021] Furthermore, the filter membrane is a polyvinylidene fluoride membrane with a pore size of 0.45 μm.

[0022] Furthermore, the freeze-drying conditions are: -50°C, vacuum degree <10Pa.

[0023] Furthermore, the preparation process of low molecular weight sodium hyaluronate is as follows: the dissolved oxygen during fermentation is maintained at 40%-60% of the saturated dissolved oxygen; the addition amount of glucose is controlled at 30-50 g / L, the addition amount of yeast extract powder is controlled at 5-10 g / L, and the addition amount of peptone is controlled at 5-10 g / L.

[0024] Furthermore, the preparation process of medium molecular weight sodium hyaluronate is as follows: the dissolved oxygen during fermentation is maintained at 20%-40% of the saturated dissolved oxygen; the addition amount of glucose is controlled at 20-30 g / L, the addition amount of yeast extract powder is controlled at 3-5 g / L, and the addition amount of peptone is controlled at 3-5 g / L.

[0025] Furthermore, the preparation process of high molecular weight sodium hyaluronate is as follows: the dissolved oxygen during fermentation is maintained at 10%-20% of the saturated dissolved oxygen; the addition amount of glucose is controlled at 10-20 g / L, the addition amount of yeast extract powder is controlled at 1-3 g / L, and the addition amount of peptone is controlled at 1-3 g / L.

[0026] Furthermore, the collagen extraction method comprises the following steps: S3-1 gene design and synthesis: Obtain a verified natural collagen gene template from a database; chemically synthesize the target gene using a DNA synthesizer; clone the target gene into an expression vector and add a His tag to simplify subsequent purification; S3-2 gene introduction technology: electroporation is used to introduce recombinant plasmids into host cells; S3-3 expression and fermentation: Pichia pastoris was induced with methanol, and Escherichia coli was induced with IPTG; the induction time was 48-72 hours, and the fermentation time was 72-96 hours; S3-4 separation and purification process: centrifugation at 8000-12000 rpm for 10-20 minutes to remove bacterial fragments, ultrafiltration and concentration of the supernatant after centrifugation to remove small molecular impurities; affinity chromatography and ion exchange chromatography; S3-5 Preparation of final product: Collagen powder was prepared by vacuum freeze drying.

[0027] Furthermore, the natural collagen genes include but are not limited to: COL1A1, COL3A1, and COL4A1.

[0028] Furthermore, the host cell is selected from one of Escherichia coli and Pichia pastoris.

[0029] Furthermore, the expression vector is selected from one of pET28a, pGEX, pPICZαA, and pPIC9K.

[0030] Furthermore, the high voltage pulse of the electroporation method is 1.5-2.5 kV.

[0031] Furthermore, the methanol induction concentration was 0.5-1%, and the IPTG induction concentration was 0.1-1 mM.

[0032] Furthermore, the fermentation temperature of Pichia pastoris is 28-30°C, and the fermentation temperature of Escherichia coli is 34-38°C.

[0033] Furthermore, the molecular weight of collagen is controlled by ultrafiltration.

[0034] Furthermore, the preparation process of small molecule collagen: during ultrafiltration, ultrafiltration membranes with a molecular weight cutoff of 2kDa and 5kDa are selected to retain the molecular weight in the target range.

[0035] Furthermore, the preparation process of macromolecular collagen: during ultrafiltration, ultrafiltration membranes with a molecular weight cutoff of 80 kDa and 100 kDa are selected to retain the molecular weight in the target range.

[0036] Furthermore, a nickel column is selected for affinity chromatography, and the eluent is 150-300 mM imidazole; a cation exchange column is selected for ion exchange chromatography, the column material is SP Sepharose HP, and the eluent is 150-250 mM NaCl.

[0037] Furthermore, the conditions for vacuum freeze drying are: -50°C, vacuum degree <10Pa.

[0038] Compared with the existing technology, the beneficial effects achieved by the present invention are: the ingredients of the present invention are streamlined and safe, reducing the risk of skin irritation. The formula contains only sodium hyaluronate, collagen and solvents, and does not add preservatives, fragrances, pigments and other irritating ingredients. It is especially suitable for sensitive skin and significantly reduces adverse reactions such as allergies.

[0039] The formula of the present invention adopts a compound molecular weight for synergistic effect: sodium hyaluronate is compounded with low, medium and high molecular weights. The high molecular weight forms a water-locking film on the skin surface, while the medium and low molecular weights penetrate deep into the dermis to moisturize, achieving multi-level hydration; collagens of different chain lengths act on the collagen fiber network of the dermis and the epidermal stratum corneum respectively, and the compound system achieves a cross-level skin care effect. The sodium hyaluronate of the present invention is produced by a fermentation method, crudely extracted with a cationic resin, degraded by proteases / nucleases for impurities, adsorbed on activated carbon, and purified by anion exchange resin, and freeze-dried to retain activity, with a purity of >98%. The collagen is prepared by a bioengineering method combined with a fermentation method, with ultrafiltration controlling the molecular weight and affinity chromatography and ion exchange chromatography for impurity removal, with a purity of >95%, retaining the natural triple helix structure and enhancing the biological activity by more than 30%.

[0040] The fiber membrane prepared by electrospinning in the present invention has a small fiber diameter and dissolves instantly when it comes into contact with water, forming a local high-concentration active substance environment with high permeability, allowing large molecular active substances to quickly reach the dermis and be absorbed by the skin; the fiber membrane has a high porosity, which guides the active functional ingredients and can provide a larger surface area for the penetration of the functional ingredients, thereby improving the penetration depth and breadth; the fiber membrane processes the difficult-to-absorb natural high-molecular active materials into extremely fine fibers for storage, does not contain water, and can maintain the activity of the functional ingredients for a long time and continuously penetrate.

[0041] In summary, the present invention solves the problems of traditional facial mask ingredient irritation, active ingredient loss, and unstable fiber performance through streamlined ingredient design, efficient purification process and electrospinning technology, achieves synergistic optimization of high safety, high activity and high fit, and significantly improves the moisturizing effect and user experience of the facial mask. DETAILED DESCRIPTION

[0042] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0043] In the following specific embodiments: the "parts" mentioned below are all parts by mass, unless otherwise specified; Low molecular weight sodium hyaluronate has a molecular weight of 5-10kDa; medium molecular weight sodium hyaluronate has a molecular weight of 200-800kDa; high molecular weight sodium hyaluronate has a molecular weight of 1000-2000kDa.

[0044] The molecular weight of small molecule collagen is 20-80kDa; the molecular weight of large molecule collagen is 80-100kDa.

[0045] The cationic resin used is hexadecyltrimethylammonium bromide, CAS No. 57-09-0; the anionic resin used is diethylaminoethyl cellulose, CAS No. 9013-34-7; and the filter membrane used is a polyvinylidene fluoride membrane with a pore size of 0.45 μm, CAS No. 24937-79-9.

[0046] The activated carbon is powdered activated carbon with the brand name YX-28 and a particle size of 1-2 mm.

[0047] The freeze-drying conditions are: -50°C, vacuum degree <10Pa.

[0048] The fermentation conditions for low molecular weight sodium hyaluronate are as follows: temperature of 37°C, dissolved oxygen maintained at 40% of saturated dissolved oxygen; the culture medium formula in the stainless steel fermenter is: 30g / L glucose, 5g / L yeast extract powder, 5g / L peptone, 0.8g / L MgSO4·7H2O, and 1.5g / L KH2PO4.

[0049] The fermentation conditions for medium molecular weight sodium hyaluronate are as follows: temperature of 37°C, dissolved oxygen maintained at 20% of saturated dissolved oxygen; the culture medium formula in the stainless steel fermenter is: 20g / L glucose, 3g / L yeast extract powder, 3g / L peptone, 0.8g / L MgSO4·7H2O, and 1.5g / L KH2PO4.

[0050] The fermentation conditions for high molecular weight sodium hyaluronate are as follows: temperature of 37°C, dissolved oxygen maintained at 10% of saturated dissolved oxygen; the culture medium formula in the stainless steel fermenter is: 10 g / L glucose, 1 g / L yeast extract powder, 1 g / L peptone, 0.8 g / L MgSO4·7H2O, and 1.5 g / L KH2PO4.

[0051] Preparation process of small molecule collagen: During ultrafiltration, select ultrafiltration membranes with a molecular weight cutoff of 2kDa and 5kDa to retain the molecular weight in the target range; Preparation process of large molecule collagen: During ultrafiltration, select ultrafiltration membranes with a molecular weight cutoff of 80kDa and 100kDa to retain the molecular weight in the target range.

[0052] Example 1: A method for preparing a fiber mask containing sodium hyaluronate and collagen, comprising the following steps: S1-1 Spinning solution preparation: Sodium hyaluronate, collagen, and solvent are added in sequence according to the formula, and stirred at 200 rpm for 4 hours to obtain a mixed solution; the mixed solution is filtered through a 500-mesh filter cloth to obtain a spinning solution; the spinning solution comprises: 5 parts sodium hyaluronate, 1 part collagen, and 94 parts solvent; the sodium hyaluronate comprises: 25 parts low molecular weight sodium hyaluronate, 35 parts medium molecular weight sodium hyaluronate, and 40 parts high molecular weight sodium hyaluronate; the collagen comprises: 70 parts small molecular weight collagen and 30 parts large molecular weight collagen; the solvent is a mixture of deionized water and n-butanol in a volume ratio of 2:1; S1-2 Electrospinning: Inject the spinning solution into the syringe and connect it to the peristaltic pump and air pump; start the peristaltic pump, air pump and high-voltage power supply, and continue spinning for 60 minutes; after the spinning is completed, remove the fiber membrane on the receiving device, cut it according to the mask mold, and trim the edges to obtain a fiber mask; the electrospinning conditions are: negative pressure 45kPa, temperature 25℃, humidity 25%, spinning rate 0.2L / h, receiving distance 50cm, non-woven fiber membrane thickness of 0.2mm, and spinning solution propulsion rate of 0.2L / h.

[0053] The method for extracting sodium hyaluronate comprises the following steps: S2-1 bacterial culture: Streptococcus was inoculated into liquid culture medium and cultured at 30°C and 150 rpm for 12 hours. 600 When the value reaches 0.7, the seed solution is obtained; the composition of the liquid culture medium is: 30g / L glucose, 5g / L yeast extract powder, 5g / L peptone, 0.8g / L MgSO4·7H2O, 1.5g / L KH2PO4; S2-2 Fermentation: The seed solution was inoculated into a stainless steel fermenter at a 10% inoculum size for fermentation for 24 hours. S2-3 crude extraction: The fermentation broth was collected and centrifuged at 8000 rpm for 10 minutes, and the supernatant was collected. 2 volumes of anhydrous ethanol were added to the supernatant, stirred, and allowed to stand for 2 hours. The hyaluronic acid precipitate was collected by centrifugation at 8000 rpm for 10 minutes. Impurity cations were removed by cationic resin, and then eluted with NaCl solution to replace the sodium salt form to produce sodium hyaluronate. S2-4 purification: add trypsin at 0.5 wt% of the mass of sodium hyaluronate, incubate at 45°C for 2 hours, add deoxyribonuclease at 0.5 wt% of the mass of sodium hyaluronate, ribonuclease at 0.5 wt% of the mass of sodium hyaluronate, and activated carbon at 0.5 wt% of the mass of sodium hyaluronate, and stir for 30 minutes; obtain a purified solution by anion exchange resin; filter the purified solution through a 0.45 μm sterilizing filter membrane, aliquot, and freeze-dry. By adjusting the dissolved oxygen concentration during fermentation and the concentration of the culture medium, sodium hyaluronate with different molecular weights can be obtained.

[0054] The method for extracting collagen comprises the following steps: S3-1‌ Gene Design and Synthesis: The natural collagen gene COL1A1 was obtained from a database; the target gene was chemically synthesized using a DNA synthesizer; the target gene was cloned into the expression vector pET28a, and a His tag was added to simplify subsequent purification; S3-2 gene introduction technology: The recombinant plasmid is introduced into E. coli BL21 by electroporation; the high voltage pulse of the electroporation method is 1.5kV; S3-3 expression and fermentation: Induction with 0.5 mM IPTG was performed for 48 hours, followed by fermentation at 37°C for 72 hours; S3-4 separation and purification process: Centrifuge at 8000 rpm for 15 minutes to remove bacterial debris. The supernatant is then concentrated by ultrafiltration to remove small molecule impurities. Affinity chromatography and ion exchange chromatography are then performed. Affinity chromatography uses a nickel column with 200 mM imidazole as the eluent. Ion exchange chromatography uses a cation exchange column with SP Sepharose HP as the eluent and 250 mM NaCl as the eluent. S3-5 Preparation of final product: Collagen powder was prepared by vacuum freeze drying.

[0055] Example 2: A method for preparing a fiber mask containing sodium hyaluronate and collagen, comprising the following steps: S1-1 Spinning solution preparation: Sodium hyaluronate, collagen, and solvent are added in sequence according to the formula, and stirred at 300 rpm for 5 hours to obtain a mixed solution; the mixed solution is filtered through a 500-mesh filter cloth to obtain a spinning solution; the spinning solution comprises: 2 parts sodium hyaluronate, 0.1 parts collagen, and 97.9 parts solvent; the sodium hyaluronate comprises: 20 parts low molecular weight sodium hyaluronate, 30 parts medium molecular weight sodium hyaluronate, and 50 parts high molecular weight sodium hyaluronate; the collagen comprises: 60 parts small molecular weight collagen and 40 parts large molecular weight collagen; the solvent is a mixture of deionized water and propanol in a volume ratio of 1:1; S1-2 Electrospinning: Inject the spinning solution into the syringe and connect it to the peristaltic pump and air pump; start the peristaltic pump, air pump and high-voltage power supply, and continue spinning for 120 minutes; after the spinning is completed, remove the fiber membrane on the receiving device, cut it according to the mask mold, and trim the edges to obtain a fiber mask; the electrospinning conditions are: negative pressure 60kPa, temperature 30℃, humidity 30%, spinning rate 0.3L / h, receiving distance 50cm, non-woven fiber membrane thickness of 0.4mm, and spinning solution propulsion rate of 0.3L / h.

[0056] The method for extracting sodium hyaluronate comprises the following steps: S2-1 bacterial culture: Acidogenic bacteria were inoculated into a liquid culture medium containing glucose, yeast extract powder, and peptone, and cultured at 34°C and 175 rpm for 18 hours. 600 When the value reaches 0.6, the seed solution is obtained; the composition of the liquid culture medium is: 10g / L glucose, 1g / L yeast extract powder, 1g / L peptone, 0.5g / L MgSO4·7H2O, 1g / L KH2PO4; S2-2 Fermentation: The seed solution was inoculated into a stainless steel fermenter at a 15% inoculum size for fermentation for 36 hours. S2-3 crude extraction: The fermentation broth was collected and centrifuged at 9000 rpm for 12 minutes, and the supernatant was collected. 2.5 volumes of anhydrous ethanol were added to the supernatant, stirred, and allowed to stand for 2.5 hours. The hyaluronic acid precipitate was then centrifuged at 9000 rpm for 15 minutes, and the hyaluronic acid precipitate was collected. Impurity cations were removed by cationic resin, and then eluted with NaCl solution to replace the sodium salt form to produce sodium hyaluronate. S2-4 purification: add trypsin at 0.7 wt% of the mass of sodium hyaluronate, incubate at 45°C for 2 hours, add deoxyribonuclease at 0.7 wt% of the mass of sodium hyaluronate, ribonuclease at 0.7 wt% of the mass of sodium hyaluronate, and activated carbon at 0.7 wt% of the mass of sodium hyaluronate, and stir for 40 minutes; obtain a purified solution by anion exchange resin; filter the purified solution through a 0.45 μm sterilizing filter membrane, aliquot, and freeze-dry. By adjusting the dissolved oxygen concentration during fermentation and the concentration of the culture medium, sodium hyaluronate with different molecular weights can be obtained.

[0057] The method for extracting collagen comprises the following steps: S3-1 gene design and synthesis: The natural collagen gene COL3A1 was obtained from a database; the target gene was chemically synthesized using a DNA synthesizer; the target gene was cloned into the expression vector pGEX, and a His tag was added to simplify subsequent purification; S3-2 gene introduction technology: The recombinant plasmid is introduced into E. coli BL21 by electroporation; the high voltage pulse of the electroporation method is 2kV; S3-3 expression and fermentation: 1 mM IPTG was used for induction. After 60 hours of induction, the cells were fermented at 34°C for 72 hours. S3-4 separation and purification process: Centrifuge at 10,000 rpm for 20 minutes to remove bacterial debris. The supernatant is then concentrated by ultrafiltration to remove small molecule impurities. Affinity chromatography and ion exchange chromatography are then performed. Affinity chromatography uses a nickel column with 250 mM imidazole as the eluent. Ion exchange chromatography uses a cation exchange column with SP Sepharose HP as the eluent and 200 mM NaCl as the eluent. S3-5 Preparation of final product: Collagen powder was prepared by vacuum freeze drying.

[0058] Example 3: A method for preparing a fiber mask containing sodium hyaluronate and collagen, comprising the following steps: S1-1 Spinning solution preparation: Sodium hyaluronate, collagen, and solvent are added in sequence according to the formula, and stirred at 400 rpm for 6 hours to obtain a mixed solution; the mixed solution is filtered through a 500-mesh filter cloth to obtain a spinning solution; the spinning solution comprises: 10 parts sodium hyaluronate, 2 parts collagen, and 88 parts solvent; the sodium hyaluronate comprises: 30 parts low molecular weight sodium hyaluronate, 50 parts medium molecular weight sodium hyaluronate, and 20 parts high molecular weight sodium hyaluronate; the collagen comprises: 80 parts small molecular weight collagen and 20 parts large molecular weight collagen; the solvent is a mixture of deionized water and 1,2-butanediol in a volume ratio of 1.5:1; S1-2 Electrospinning: Inject the spinning solution into the syringe and connect it to the peristaltic pump and air pump; start the peristaltic pump, air pump and high-voltage power supply, and continue spinning for 100 minutes; after the spinning is completed, remove the fiber membrane on the receiving device, cut it according to the mask mold, and trim the edges to obtain a fiber mask; the conditions for electrospinning are: negative pressure 30kPa, temperature 20℃, humidity 20%, spinning rate 0.4L / h, receiving distance 50cm, non-woven fiber membrane thickness of 0.5mm, and spinning solution propulsion rate of 0.4L / h.

[0059] The method for extracting sodium hyaluronate comprises the following steps: S2-1 bacterial culture: The fermentation yeast was inoculated into a liquid culture medium containing glucose, yeast extract powder, and peptone, and cultured at 37°C and 200 rpm for 24 hours. 600 When the value reaches 0.8, the seed solution is obtained; the composition of the liquid culture medium is: 50g / L glucose, 10g / L yeast extract powder, 10g / L peptone, 1g / L MgSO4·7H2O, 2g / L KH2PO4; S2-2 Fermentation: The seed solution was inoculated into a stainless steel fermenter at a 20% inoculum size for fermentation for 48 hours. S2-3 crude extraction: The fermentation broth was collected and centrifuged at 10,000 rpm for 15 minutes, and the supernatant was collected. Three volumes of anhydrous ethanol were added to the supernatant, stirred, and allowed to stand for 3 hours. The hyaluronic acid precipitate was collected by centrifugation at 10,000 rpm for 20 minutes. Impurity cations were removed by cationic resin, and then eluted with NaCl solution to replace the sodium salt form to produce sodium hyaluronate. S2-4 purification: add trypsin at 1 wt% of the mass of sodium hyaluronate, incubate at 45°C for 2 hours, add deoxyribonuclease at 1 wt% of the mass of sodium hyaluronate, ribonuclease at 1 wt% of the mass of sodium hyaluronate, and activated carbon at 1 wt% of the mass of sodium hyaluronate, and stir for 45 minutes; obtain a purified solution by anion exchange resin; filter the purified solution through a 0.45 μm sterilizing filter membrane, aliquot, and freeze-dry. By adjusting the dissolved oxygen concentration during fermentation and the concentration of the culture medium, sodium hyaluronate with different molecular weights can be obtained.

[0060] The method for extracting collagen comprises the following steps: S3-1‌ Gene Design and Synthesis: The natural collagen gene COL4A1 was obtained from a database; the target gene was chemically synthesized using a DNA synthesizer; the target gene was cloned into the expression vector pPICZαA, and a His tag was added to simplify subsequent purification; S3-2 gene introduction technology: The recombinant plasmid was introduced into Pichia pastoris GS115 by electroporation; the high voltage pulse of the electroporation method was 2.5kV; S3-3 expression and fermentation: 0.5% methanol was used for induction, and after 72 hours of induction, fermentation was carried out at 38°C for 72 hours; S3-4 separation and purification process: Centrifuge at 12,000 rpm for 10 minutes to remove bacterial debris. The supernatant was then concentrated by ultrafiltration to remove small molecule impurities. Affinity chromatography and ion exchange chromatography were performed. Affinity chromatography used a nickel column with 150 mM imidazole as the eluent. Ion exchange chromatography used a cation exchange column with SP Sepharose HP as the column material and 150 mM NaCl as the eluent. S3-5 Preparation of final product: Collagen powder was prepared by vacuum freeze drying.

[0061] Comparative Example 1: Based on Example 1, the spinning solution formula was adjusted to adjust the sodium hyaluronate compound solution to medium molecular weight sodium hyaluronate, specifically comprising the following steps: A method for preparing a fiber mask containing sodium hyaluronate and collagen comprises the following steps: S1-1 Spinning solution preparation: Sodium hyaluronate, collagen, and solvent are added in sequence according to the formula, and stirred at 200 rpm for 4 hours to obtain a mixed solution; the mixed solution is filtered through a 500-mesh filter cloth to obtain a spinning solution; the spinning solution comprises: 5 parts sodium hyaluronate, 1 part collagen, and 94 parts solvent; the sodium hyaluronate comprises: 5 parts medium molecular weight sodium hyaluronate, 1 part collagen, and 94 parts solvent; the collagen comprises: 70 parts small molecular weight collagen and 30 parts large molecular weight collagen; the solvent is a mixture of deionized water and n-butanol in a volume ratio of 2:1; S1-2 Electrospinning: Inject the spinning solution into the syringe and connect it to the peristaltic pump and air pump; start the peristaltic pump, air pump and high-voltage power supply, and continue spinning for 60 minutes; after the spinning is completed, remove the fiber membrane on the receiving device, cut it according to the mask mold, and trim the edges to obtain a fiber mask; the electrospinning conditions are: negative pressure 45kPa, temperature 25℃, humidity 25%, spinning rate 0.2L / h, receiving distance 50cm, non-woven fiber membrane thickness of 0.2mm, and spinning solution propulsion rate of 0.2L / h.

[0062] The method for extracting sodium hyaluronate comprises the following steps: S2-1 bacterial culture: Streptococcus was inoculated into liquid culture medium and cultured at 30°C and 150 rpm for 12 hours. 600 When the value reaches 0.7, the seed solution is obtained; the composition of the liquid culture medium is: 30g / L glucose, 5g / L yeast extract powder, 5g / L peptone, 0.8g / L MgSO4·7H2O, 1.5g / L KH2PO4; S2-2 Fermentation: The seed solution was inoculated into a stainless steel fermenter at a 10% inoculum size for fermentation for 24 hours. S2-3 crude extraction: The fermentation broth was collected and centrifuged at 8000 rpm for 10 minutes, and the supernatant was collected. 2 volumes of anhydrous ethanol were added to the supernatant, stirred, and allowed to stand for 2 hours. The hyaluronic acid precipitate was collected by centrifugation at 8000 rpm for 10 minutes. Impurity cations were removed by cationic resin, and then eluted with NaCl solution to replace the sodium salt form to produce sodium hyaluronate. S2-4 purification: add trypsin at 0.5 wt% of the mass of sodium hyaluronate, incubate at 45°C for 2 hours, add deoxyribonuclease at 0.5 wt% of the mass of sodium hyaluronate, ribonuclease at 0.5 wt% of the mass of sodium hyaluronate, and activated carbon at 0.5 wt% of the mass of sodium hyaluronate, and stir for 30 minutes; obtain a purified solution by anion exchange resin; filter the purified solution through a 0.45 μm sterilizing filter membrane, aliquot, and freeze-dry. By adjusting the dissolved oxygen concentration during fermentation and the concentration of the culture medium, sodium hyaluronate with different molecular weights can be obtained.

[0063] The method for extracting collagen comprises the following steps: S3-1‌ Gene Design and Synthesis: The natural collagen gene COL1A1 was obtained from a database; the target gene was chemically synthesized using a DNA synthesizer; the target gene was cloned into the expression vector pET28a, and a His tag was added to simplify subsequent purification; S3-2 gene introduction technology: The recombinant plasmid is introduced into E. coli BL21 by electroporation; the high voltage pulse of the electroporation method is 1.5kV; S3-3 expression and fermentation: Induction with 0.5 mM IPTG was performed for 48 hours, followed by fermentation at 37°C for 72 hours; S3-4 separation and purification process: Centrifuge at 8000 rpm for 15 minutes to remove bacterial debris. The supernatant is then concentrated by ultrafiltration to remove small molecule impurities. Affinity chromatography and ion exchange chromatography are then performed. Affinity chromatography uses a nickel column with 200 mM imidazole as the eluent. Ion exchange chromatography uses a cation exchange column with SP Sepharose HP as the eluent and 250 mM NaCl as the eluent. S3-5 Preparation of final product: Collagen powder was prepared by vacuum freeze drying.

[0064] Comparative Example 2: Based on Example 1, the spinning solution formula was adjusted to adjust the collagen compound solution to small molecule collagen, specifically comprising the following steps: A method for preparing a fiber mask containing sodium hyaluronate and collagen comprises the following steps: S1-1 Spinning solution preparation: Sodium hyaluronate, collagen, and solvent are added in sequence according to the formula and stirred at 200 rpm for 4 hours to obtain a uniform solution; the mixed solution is filtered through a 500-mesh filter cloth to obtain a spinning solution; the spinning solution comprises: 5 parts sodium hyaluronate, 1 part low-molecule collagen, and 94 parts solvent; the sodium hyaluronate comprises: 25 parts low-molecular-weight sodium hyaluronate and 35 parts medium-molecular-weight sodium hyaluronate; the solvent is a mixture of deionized water and n-butanol in a volume ratio of 2:1; S1-2 Electrospinning: Inject the spinning solution into the syringe and connect it to the peristaltic pump and air pump; start the peristaltic pump, air pump and high-voltage power supply, and continue spinning for 60 minutes; after the spinning is completed, remove the fiber membrane on the receiving device, cut it according to the mask mold, and trim the edges to obtain a fiber mask; the electrospinning conditions are: negative pressure 45kPa, temperature 25℃, humidity 25%, spinning rate 0.2L / h, receiving distance 50cm, non-woven fiber membrane thickness of 0.2mm, and spinning solution propulsion rate of 0.2L / h.

[0065] The method for extracting sodium hyaluronate comprises the following steps: S2-1 bacterial culture: Streptococcus was inoculated into liquid culture medium and cultured at 30°C and 150 rpm for 12 hours. 600 When the value reaches 0.7, the seed solution is obtained; the composition of the liquid culture medium is: 30g / L glucose, 5g / L yeast extract powder, 5g / L peptone, 0.8g / L MgSO4·7H2O, 1.5g / L KH2PO4; S2-2 Fermentation: The seed solution was inoculated into a stainless steel fermenter at a 10% inoculum size for fermentation for 24 hours. S2-3 crude extraction: The fermentation broth was collected and centrifuged at 8000 rpm for 10 minutes, and the supernatant was collected. 2 volumes of anhydrous ethanol were added to the supernatant, stirred, and allowed to stand for 2 hours. The hyaluronic acid precipitate was collected by centrifugation at 8000 rpm for 10 minutes. Impurity cations were removed by cationic resin, and then eluted with NaCl solution to replace the sodium salt form to produce sodium hyaluronate. S2-4 purification: add trypsin at 0.5 wt% of the mass of sodium hyaluronate, incubate at 45°C for 2 hours, add deoxyribonuclease at 0.5 wt% of the mass of sodium hyaluronate, ribonuclease at 0.5 wt% of the mass of sodium hyaluronate, and activated carbon at 0.5 wt% of the mass of sodium hyaluronate, and stir for 30 minutes; obtain a purified solution by anion exchange resin; filter the purified solution through a 0.45 μm sterilizing filter membrane, aliquot, and freeze-dry. By adjusting the dissolved oxygen concentration during fermentation and the concentration of the culture medium, sodium hyaluronate with different molecular weights can be obtained.

[0066] The method for extracting collagen comprises the following steps: S3-1‌ Gene Design and Synthesis: The natural collagen gene COL1A1 was obtained from a database; the target gene was chemically synthesized using a DNA synthesizer; the target gene was cloned into the expression vector pET28a, and a His tag was added to simplify subsequent purification; S3-2 gene introduction technology: The recombinant plasmid is introduced into E. coli BL21 by electroporation; the high voltage pulse of the electroporation method is 1.5kV; S3-3 expression and fermentation: Induction with 0.5 mM IPTG was performed for 48 hours, followed by fermentation at 37°C for 72 hours; S3-4 separation and purification process: Centrifuge at 8000 rpm for 15 minutes to remove bacterial debris. The supernatant is then concentrated by ultrafiltration to remove small molecule impurities. Affinity chromatography and ion exchange chromatography are then performed. Affinity chromatography uses a nickel column with 200 mM imidazole as the eluent. Ion exchange chromatography uses a cation exchange column with SP Sepharose HP as the eluent and 250 mM NaCl as the eluent. S3-5 Preparation of final product: Collagen powder was prepared by vacuum freeze drying.

[0067] Comparative Example 3: Based on Example 1, the preparation method of the fiber mask was adjusted by changing the electrospinning to wet spinning, specifically comprising the following steps: A method for preparing a fiber mask containing sodium hyaluronate and collagen comprises the following steps: S1-1 Spinning solution preparation: Sodium hyaluronate, collagen, and solvent are added in sequence according to the formula, and stirred at 200 rpm for 4 hours to obtain a mixed solution; the mixed solution is filtered through a 500-mesh filter cloth to obtain a spinning solution; the spinning solution comprises: 5 parts sodium hyaluronate, 1 part collagen, and 94 parts solvent; the sodium hyaluronate comprises: 25 parts low molecular weight sodium hyaluronate, 35 parts medium molecular weight sodium hyaluronate, and 40 parts high molecular weight sodium hyaluronate; the collagen comprises: 70 parts small molecular weight collagen and 30 parts large molecular weight collagen; the solvent is a mixture of deionized water and n-butanol in a volume ratio of 2:1; S1-2 Wet Spinning: The spinning solution is transported to a spinning machine for wet spinning. The spinning solution is extruded through a spinneret with a pore size of 0.075 mm to form a thin stream. After entering the coagulation bath, solvent-coagulant double diffusion occurs to form nascent fibers. The spinning speed is 50 m / min. 70% ethanol is used as the coagulation bath at a temperature of 20°C. The distance between the coagulation bath liquid surface and the spinneret is 10 cm. S1-3 Removal of organic solvent: Place the fiber membrane in a fume hood for 1 hour; Place the fiber membrane that has undergone preliminary ventilation and volatilization in an oven, set the temperature to 40°C, and place it for 2 hours; Soak the fiber membrane in an appropriate amount of ethanol for 1 hour, stirring appropriately during the period to allow the organic solvent to fully dissolve in the replacement solvent, remove the fiber membrane, and air dry it at room temperature again to remove the replacement solvent; Place the fiber membrane that has been treated previously in a vacuum drying oven, evacuate to -0.08 MPa, set the temperature to 40°C, and maintain for 1 hour to further remove the residual organic solvent; S1-4 Moisturizing pretreatment: Immerse the fiber membrane in a deionized water solution containing 10% glycerol for 5 minutes, remove and drain; S1-5 Cross-linking treatment: The fiber membrane obtained in the previous step is irradiated with ultraviolet light to enhance the tensile strength of the fiber membrane and obtain a fiber mask. The conditions of ultraviolet radiation are: wavelength of 254 nm, irradiation time of 10 minutes, and distance of 35 cm.

[0068] The method for extracting sodium hyaluronate comprises the following steps: S2-1 bacterial culture: Streptococcus was inoculated into liquid culture medium and cultured at 30°C and 150 rpm for 12 hours. 600 When the value reaches 0.7, the seed solution is obtained; the composition of the liquid culture medium is: 30g / L glucose, 5g / L yeast extract powder, 5g / L peptone, 0.8g / L MgSO4·7H2O, 1.5g / L KH2PO4; S2-2 Fermentation: The seed solution was inoculated into a stainless steel fermenter at a 10% inoculum size for fermentation for 24 hours. S2-3 crude extraction: The fermentation broth was collected and centrifuged at 8000 rpm for 10 minutes, and the supernatant was collected. 2 volumes of anhydrous ethanol were added to the supernatant, stirred, and allowed to stand for 2 hours. The hyaluronic acid precipitate was collected by centrifugation at 8000 rpm for 10 minutes. Impurity cations were removed by cationic resin, and then eluted with NaCl solution to replace the sodium salt form to produce sodium hyaluronate. S2-4 purification: add trypsin at 0.5 wt% of the mass of sodium hyaluronate, incubate at 45°C for 2 hours, add deoxyribonuclease at 0.5 wt% of the mass of sodium hyaluronate, ribonuclease at 0.5 wt% of the mass of sodium hyaluronate, and activated carbon at 0.5 wt% of the mass of sodium hyaluronate, and stir for 30 minutes; obtain a purified solution by anion exchange resin; filter the purified solution through a 0.45 μm sterilizing filter membrane, aliquot, and freeze-dry. By adjusting the dissolved oxygen concentration during fermentation and the concentration of the culture medium, sodium hyaluronate with different molecular weights can be obtained.

[0069] The method for extracting collagen comprises the following steps: S3-1‌ Gene Design and Synthesis: The natural collagen gene COL1A1 was obtained from a database; the target gene was chemically synthesized using a DNA synthesizer; the target gene was cloned into the expression vector pET28a, and a His tag was added to simplify subsequent purification; S3-2 gene introduction technology: The recombinant plasmid is introduced into E. coli BL21 by electroporation; the high voltage pulse of the electroporation method is 1.5kV; S3-3 expression and fermentation: Induction with 0.5 mM IPTG was performed for 48 hours, followed by fermentation at 37°C for 72 hours; S3-4 separation and purification process: Centrifuge at 8000 rpm for 15 minutes to remove bacterial debris. The supernatant is then concentrated by ultrafiltration to remove small molecule impurities. Affinity chromatography and ion exchange chromatography are then performed. Affinity chromatography uses a nickel column with 200 mM imidazole as the eluent. Ion exchange chromatography uses a cation exchange column with SP Sepharose HP as the eluent and 250 mM NaCl as the eluent. S3-5 Preparation of final product: Collagen powder was prepared by vacuum freeze drying.

[0070] Experiment: 120 people aged 18-50 years old, half male and half female, were randomly divided into 6 groups. Each group used the facial masks of Examples 1-3 and Comparative Examples 1-3 respectively. No drugs or cosmetics unrelated to the experiment were applied during the experiment.

[0071] Before the experiment, the subjects' skin moisture content, skin brightness and skin elasticity were tested as baseline values. After cleansing their faces every night, each group of subjects used the corresponding fiber mask on their faces.

[0072] One hour after using the fiber mask, skin moisture was measured using the MY-808S Skin Moisture Analyzer; skin brightness was measured using the VISIA-CR Facial Skin Image Analyzer; and skin elasticity was measured using the MPA580 Skin Elasticity Tester. Data are averages for all data within the same test group. The experimental results are shown in Tables 1, 2, and 3.

[0073] Table 1 Skin moisture content test results

[0074] Conclusion: As shown in Table 1, the skin moisture content of the subjects increased with the use time of the fiber masks of Examples 1-3 and Comparative Examples 1-3. However, a comparison of the data shows that in Comparative Example 1, where the compounded sodium hyaluronate was adjusted to medium molecular weight sodium hyaluronate, the increase in skin moisture content was lower than that of the Example; in Comparative Example 2, where the compounded collagen was adjusted to low molecular weight collagen, the increase in skin moisture content was lower than that of the Example; and in Comparative Example 3, where the electrospinning was adjusted to wet spinning, the increase in skin moisture content was lower than that of the Example. This fully demonstrates that the fiber mask of the present invention can effectively utilize the efficacy of each component of the hydrogel matrix, achieving comprehensive moisturizing care by reducing transepidermal water loss, enhancing internal cellular water circulation, and strengthening the surface water barrier.

[0075] Table 2 Skin brightness test results

[0076] As shown in Table 2, the skin brightness of the subjects increased with the use time of the fiber masks of Examples 1-3 and Comparative Examples 1-3. However, a comparison of the data shows that in Comparative Example 1, the increase in skin brightness was lower than that of the Example, as the sodium hyaluronate was adjusted to a medium molecular weight sodium hyaluronate; in Comparative Example 2, the increase in skin brightness was lower than that of the Example, as the collagen was adjusted to a low molecular weight collagen; and in Comparative Example 3, the increase in skin brightness was lower than that of the Example, as the electrospinning was adjusted to a wet spinning method. This indicates that the fiber mask of the present invention has a highly effective whitening effect.

[0077] Table 3 Skin elasticity test results

[0078] As can be seen from the results in Table 3, with the extension of the use time, the skin elasticity of the subjects increased after using the fiber masks of Examples 1-3 and Comparative Examples 1-3. However, data comparison shows that Comparative Example 1 adjusts the compounded sodium hyaluronate to medium molecular weight sodium hyaluronate, and the increase in skin elasticity is lower than that of the embodiment; Comparative Example 2 adjusts the compounded collagen to low molecular weight collagen, and the increase in skin elasticity is lower than that of the embodiment; Comparative Example 3 adjusts electrospinning to wet spinning, and the increase in skin elasticity is lower than that of the embodiment. After using the fiber mask of the present invention, the skin elasticity of the subjects increased, indicating that the fiber mask of the present invention improves skin nutrient metabolism, maintains skin elasticity, and repairs epidermal tissue at the same time, thereby achieving multiple effects of moisturizing, anti-wrinkle and anti-aging.

[0079] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations that come within the meaning and range of equivalents of the claims be embraced therein.

Claims

1. A fiber mask containing sodium hyaluronate and collagen, characterized by: The spinning solution is prepared by electrostatic spinning, wherein the spinning solution comprises the following components in parts by weight: 2-10 parts of sodium hyaluronate, 0.1-2 parts of collagen, and 88-97.9 parts of solvent.

2. The fiber mask containing sodium hyaluronate and collagen according to claim 1, characterized in that: The sodium hyaluronate is compounded by sodium hyaluronates of different molecular weights, specifically including the following components in parts by weight: low molecular weight sodium hyaluronate: 20-30 parts; medium molecular weight sodium hyaluronate: 30-50 parts; high molecular weight sodium hyaluronate: 20-50 parts.

3. The fiber mask containing sodium hyaluronate and collagen according to claim 2, characterized in that: The collagen is compounded by collagens of different molecular weights, and specifically includes the following components in parts by weight: small molecule collagen: 60-80 parts; large molecule collagen: 20-40 parts.

4. The fiber mask containing sodium hyaluronate and collagen according to claim 3, characterized in that: The solvent formula is: deionized water and organic solvent are compounded in a volume ratio of 1-2:

1.

5. The fiber facial mask containing sodium hyaluronate and collagen according to claim 4, characterized in that: The organic solvent is selected from one of methyl ethyl ketone, N-methyl pyrrolidone, 1,3-propylene glycol, 1,2-butanediol, 1,4-butanediol, 2,3-butanediol, diacetone alcohol, acetone, n-butanol, and propanol.

6. A method for preparing a fiber facial mask containing sodium hyaluronate and collagen, characterized in that: The method for making the fiber mask comprises the following steps: S1-1 Spinning solution preparation: Sodium hyaluronate, collagen, and solvent are added in sequence according to the formula, and stirred at 200-400 rpm for 4-6 hours to obtain a mixed solution; the mixed solution is filtered through a 500-mesh filter cloth to obtain a spinning solution; S1-2 Electrospinning: Inject the spinning solution into the syringe and connect it to the peristaltic pump and air pump; start the peristaltic pump, air pump and high-voltage power supply, and continue spinning for 60-120 minutes; after the spinning is completed, remove the fiber membrane from the receiving device, cut it according to the mask mold, and trim the edges to obtain a fiber mask.

7. The method for preparing a fiber mask containing sodium hyaluronate and collagen according to claim 6, wherein: The electrospinning conditions are: negative pressure 30-60 kPa, temperature 20-30° C., humidity 20-30%, spinning rate 0.2-0.4 L / h, and receiving distance 50 cm.

8. The method for preparing a fiber mask containing sodium hyaluronate and collagen according to claim 7, wherein: The spinning solution advancing rate is 0.2-0.4 L / h, and the fiber membrane thickness is 0.1-0.5 mm.

9. The method for preparing a fiber facial mask containing sodium hyaluronate and collagen according to claim 6, wherein: The method for extracting sodium hyaluronate comprises the following steps: S2-1 Bacterial culture: Inoculate the bacterial strain into liquid culture medium, shake and culture at 30-37°C and 150-200 rpm for 12-24 hours to obtain seed solution; S2-2 Fermentation culture: The seed liquid is inoculated into a stainless steel fermentation tank for fermentation culture, and the fermentation time is 24-48 hours; S2-3 crude extraction: The fermentation broth is collected and centrifuged at 8000-10000 rpm for 10-15 minutes, and the supernatant is collected; 2-3 volumes of anhydrous ethanol are added to the supernatant, stirred, and allowed to stand for 2-3 hours, and centrifuged at 8000-10000 rpm for 10-20 minutes to collect the hyaluronic acid precipitate; impure cations are removed by cationic resin, and then eluted with NaCl solution to replace the sodium salt form to produce sodium hyaluronate; S2-4 purification: add trypsin, incubate at 45-55°C for 2-4 hours, add deoxyribonuclease, ribonuclease and activated carbon, and stir for 30-45 minutes; pass through anion exchange resin to obtain a purified solution; filter the purified solution through a 0.45 μm sterilizing filter membrane, divide into portions and freeze-dry to obtain sodium hyaluronate.

10. The method for preparing a fiber facial mask containing sodium hyaluronate and collagen according to claim 6, characterized in that: The collagen extraction method comprises the following steps: S3-1 gene design and synthesis: Obtain a verified natural collagen gene template from a database; chemically synthesize the target gene using a DNA synthesizer; clone the target gene into an expression vector and add a His tag to simplify subsequent purification; S3-2 gene introduction technology: electroporation is used to introduce recombinant plasmids into host cells; S3-3 expression and fermentation: Pichia pastoris was induced with methanol, and Escherichia coli was induced with IPTG; the induction time was 48-72 hours, and the fermentation time was 72-96 hours; S3-4 separation and purification process: centrifuge at 8000-12000 rpm for 10-20 minutes to remove bacterial fragments, and then perform ultrafiltration and concentration of the supernatant to remove small molecule impurities; perform affinity chromatography and ion exchange chromatography; S3-5 Preparation of final product: Collagen powder was prepared by vacuum freeze drying.

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