Tightening essence spray during coagulation

Abstract

The invention relates to the technical field of cosmeceuticals, in particular to a tightening essence spray. The anti-wrinkle mask is prepared from the following components in percentage by mass: 83.56% of bifidus yeast fermentation product filtrate, 6.5% of an anti-wrinkle agent A, 3.5% of an anti-wrinkle agent B, 2.5% of a preservative, 1.5% of nicotinamide, 1% of an anti-wrinkle agent C, 1% of Wailing enzyme, 0.11% of dipotassium glycyrrhizinate, 0.11% of allantoin, 0.11% of fullerene and 0.11% of hydrolyzed collagen. The anti-wrinkle agent A contains arginine / lysine polypeptide, and the molecular weight of the anti-wrinkle agent A is 1t; the 500Da oligopeptide chain can directly penetrate through the stratum corneum, and is competitively combined with the SNAP-25 protein to block the transmission of neuromuscular signals. The anti-wrinkle agent B contains acetyl hexapeptide-8, and forms a'nerve blocking-muscle relaxing 'closed loop with the anti-wrinkle agent A by inhibiting a calcium ion channel and reducing the depth of dynamic lines, and the effect is remarkable. The nicotinamide in the invention blocks melanosome transfer, activates an AMPK pathway to promote autophagy flow, and accelerates the degradation of generated melanin. The dipotassium glycyrrhizinate inhibits COX-2 inflammatory enzyme, reduces the level of PGE2 inflammatory factors induced by ultraviolet rays, and blocks inflammatory chromaticity.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of cosmeceutics, in particular to a firming essence spray. BACKGROUND

[0002] The firming essence spray is a kind of skin care product that combines the effects of firming and anti-aging with a spray type. Through the design of concentrated essence components and atomizing nozzles, it achieves rapid absorption and targeted anti-aging effects, meeting the needs of modern women for "minimalist skincare" and "on-the-go maintenance".

[0003] Most of the anti-aging firming products on the market contain nerve signal inhibitor ingredients (such as Moritek's XEP-018 and ECST-5 technology). However, different individuals have different reactions to nerve signal inhibitor ingredients, and some people may be allergic to these ingredients, causing skin redness, itching, and skin rash. In addition, long-term or excessive use of cosmeceutics containing nerve signal inhibitor ingredients may interfere with normal nerve conduction. If these ingredients are absorbed through the skin and enter the blood circulation, they may have adverse effects on the nervous system. Although the current research is not clear, the potential risks cannot be ignored. Finally, the compatibility of nerve signal inhibitors with other ingredients in the formulation of cosmeceutics is also a challenge. Adding too much concentration not only increases the cost of research and development for enterprises, but also may not be well compounded with other ingredients, affecting product stability and effectiveness.

[0004] Therefore, there is an urgent need for a safe and effective firming essence spray. SUMMARY

[0005] The purpose of the present application is to provide a firming essence spray to solve at least one of the above technical problems.

[0006] The technical solution adopted by the present application to solve its technical problems is: A firming essence spray, which comprises the following components in percentage by mass: 83.56% of Schizochytrium limacinum ferment filtrate, 6.5% of anti-wrinkle agent A, 3.5% of anti-wrinkle agent B, 2.5% of preservative, 1.5% of nicotinamide, 1% of anti-wrinkle agent C, 1% of winter essence, 0.11% of dipotassium glycyrrhizinate, 0.11% of allantoin, 0.11% of fullers sugar, and 0.11% of hydrolyzed collagen.

[0007] Further, the preparation method of the Schizochytrium limacinum ferment filtrate comprises the following steps: S1 inoculation culture: inoculate the activated strain in logarithmic growth phase into a seed tank that has been sterilized and filled with nitrogen gas to replace air, at an inoculation amount (volume ratio) of 5%-10%, and incubate at a constant temperature for 18-24h, until the viable cell count of the seed solution is ≥109 CFU / mL, OD 600 values reach 1.2-1.5; S2 fermentation: inoculate the seed liquid in step S1 into the main fermentation tank (1000-5000L) at an inoculation amount of 3%-5%, maintain the carbon dioxide concentration in the fermentation tank at 5%-8%, and use stepwise culture control: 1) control the pH value at 6.0-6.5, the glucose concentration at 1%-2%, and the temperature at 37±1℃ for 24h in the growth stage; 2) reduce the temperature to 35℃, adjust the pH to 5.8-6.0, and limit the glucose concentration to 0.5%-1% in the active substance synthesis stage to induce the synthesis of active substances, including polysaccharides and polypeptides, by the bacteria, and the fermentation period is 60-72h; S3 cell lysis and separation of filtrate: add 0.1%-0.2% lysozyme, treat at 37℃ for 1-2h, and then sequentially pass through centrifugal separation and microfiltration to obtain the Schizochytrium sp. fermentation product filtrate.

[0008] Further, the preparation method further comprises desalination and concentration, and the specific steps are as follows: desalinate the filtrate in step S3 by nanofiltration, the nanofiltration membrane has a molecular weight cut-off of 300-500Da, retains active small molecules, and removes salts; then, use low-temperature vacuum concentration, control the temperature ≤40℃, the vacuum degree is -0.08 to -0.09MPa, and concentrate the filtrate to 1 / 5-1 / 10 of the original volume.

[0009] Further, the anti-wrinkle agent A comprises the following components by mass fraction: water 60-80 parts butanediol 1-2 parts trehalose 0.1-0.5 parts 1,2-hexanediol 1-2 parts ethylhexyl glycerin 0.1-0.3 parts arginine / lysine polypeptide 0.1-0.5 parts.

[0010] Further, the anti-wrinkle agent B comprises the following components by mass fraction: water 60-84 parts glycerol 1-3 parts octanediol 0.1-0.8 parts acetyl hexapeptide-8 0.01-0.05 parts ethylhexyl glycerin 0.01-0.15 parts.

[0011] Further, the preservative comprises 1,2-pentanediol and p-hydroxyacetophenone, and the mass ratio of 1,2-pentanediol to p-hydroxyacetophenone is (2-3):1.

[0012] Further, the quality parts composition of the dongling enzyme is as follows: Schizosaccharomyces pombe fermentation product filtrate 85-95 parts Glycerol 2-10 parts 1,2-pentanediol 1-5 parts Ophiopogon root extract 2-3.5 parts Red Ganoderma extract 0.5-2 parts 4-hydroxyacetophenone 0.05-0.3 parts.

[0013] Further, the fullerenes include zinc gluconate and fullerene, and the mass ratio is (1-6):1.

[0014] The beneficial effects of the present application are: In the present application, Schizosaccharomyces pombe filtrate and dongling enzyme are added at the same time, and the two yeasts are synergistic to achieve "micro-ecological level" anti-aging. The Schizosaccharomyces pombe filtrate provides DNA repair enzymes, beta-glucan, organic acids and other micro-ecological nutrients, directly activates the Nrf2 / ARE antioxidant pathway of keratinocytes, and the dongling enzyme inhibits the TRPV1 nerve receptor by synergistic effect of Ophiopogon root extract (steroidal saponin) and Ganoderma lucidum extract (triterpenoids), reduces the pain signal transmission of sensitive skin, and realizes "instant relief + long-term repair", solving the micro-stimulation that may be caused by high-activity yeast.

[0015] The anti-wrinkle agent A in the present application contains arginine / lysine polypeptide, and the oligopeptide chain with a molecular weight of <500 Da can directly penetrate the stratum corneum, competitively bind with SNAP-25 protein, and block nerve-muscle signal transmission. The anti-wrinkle agent B contains acetyl hexapeptide-8, which can inhibit calcium ion channels and reduce dynamic wrinkle depth, and forms a "nerve block-muscle relaxation" closed loop with the anti-wrinkle agent A, and the effect is remarkable.

[0016] In the present application, nicotinamide blocks the transfer of melanosome, activates the AMPK pathway to promote autophagy flow, and accelerates the degradation of the generated melanin. Dipotassium glycyrrhizinate inhibits COX-2 inflammatory enzymes and reduces the level of PGE2 inflammatory factors induced by ultraviolet light, thereby blocking inflammatory pigmentation.

[0017] In the present application, the hydrolyzed collagen is obtained by enzyme cutting technology to obtain collagen tripeptide (GPH) with a molecular weight of <1000 Da, which can directly stimulate fibroblast proliferation and activate the TGF-β / Smad pathway to promote type I collagen synthesis.

[0018] In the present application, trehalose forms a "glassy" protective film on the skin surface, reduces transdermal water loss, and at the same time, through hydration, makes the collagen fibers more closely arranged, realizing "instant firming + long-term collagen neogenesis".

[0019] The preparation method of the Schizochytrium ferment product filtrate in the application can efficiently produce the Schizochytrium ferment product filtrate rich in active substances by accurately controlling the fermentation conditions and the post-treatment steps; by desalting and concentration treatment, the purity and concentration of the product are improved, and the stability and application effect of the product are enhanced; the whole process design is reasonable, the operation is simple, and the application prospect is high. DETAILED DESCRIPTION

[0021] The technical solutions of the application will be described clearly and completely in combination with the embodiments. Obviously, the described embodiments are only some of the embodiments of the application, rather than all the embodiments. Based on the embodiments of the application, all other embodiments obtained by those skilled in the art without creative work belong to the protection scope of the application.

[0022] The condensation time firming essence spray in the application has the following mass percentage composition: Schizochytrium ferment product filtrate 83.56%, anti-wrinkle agent A 6.5%, anti-wrinkle agent B 3.5%, preservative 2.5%, nicotinamide 1.5%, anti-wrinkle agent C 1%, winter essence 1%, glycyrrhizinate 0.11%, allantoin 0.11%, fuller sugar 0.11%, and hydrolyzed collagen 0.11%.

[0023] The mass parts of anti-wrinkle agent A are as follows: Water 60-80 parts Butylene glycol 1-2 parts Trehalose 0.1-0.5 parts 1,2-Hexanediol 1-2 parts Ethylhexylglycerin 0.1-0.3 parts Arginine / lysine polypeptide 0.1-0.5 parts.

[0024] The mass parts of anti-wrinkle agent B are as follows: Water 60-84 parts Glycerin 1-3 parts Octanediol 0.1-0.8 parts Acetyl hexapeptide-8 0.01-0.05 parts Ethylhexylglycerin 0.01-0.15 parts.

[0025] The mass parts of winter essence are as follows: Schizochytrium ferment product filtrate 85-95 parts Glycerin 2-10 parts 1,2-Pentanediol 1-5 parts Radix ophiopogonis extract 2-3.5 parts Red ganoderma extract 0.5-2 parts p-Hydroxyacetophenone 0.05-0.3 parts.

[0026] Example 1 A condensed firming essence spray has the following mass percentage composition: Schizochytrium ferment filtrate 83.56%, anti-wrinkle agent A 6.5%, anti-wrinkle agent B 3.5%, preservative 2.5%, nicotinamide 1.5%, anti-wrinkle agent C 1%, Dongling enzyme 1%, dipotassium glycyrrhizinate 0.11%, allantoin 0.11%, fuller sugar 0.11%, hydrolyzed collagen 0.11%.

[0027] Anti-wrinkle agent A has the following mass parts composition: Water 60 parts Butylene glycol 1 part Trehalose 0.1 part 1,2-Hexanediol 1 part Ethylhexylglycerin 0.1 part Arginine / lysine polypeptide 0.1 part.

[0028] Anti-wrinkle agent B has the following mass parts composition: Water 60 parts Glycerin 1 part Octanediol 0.1 part Acetyl hexapeptide-8 0.01 part Ethylhexylglycerin 0.01 part.

[0029] The preservative includes 1,2-pentanediol and p-hydroxyacetophenone, and the mass ratio of 1,2-pentanediol to p-hydroxyacetophenone is 2:1.

[0030] Dongling enzyme has the following mass parts composition: Schizochytrium ferment filtrate 85 parts Glycerin 2 parts 1,2-Pentanediol 1 part Radix ophiopogonis extract 2 parts Red ganoderma extract 0.5 part p-Hydroxyacetophenone 0.05 part.

[0031] Fuller sugar includes zinc gluconate and fullerene, and the mass ratio is 1:1.

[0032] The preparation method of the Schizochytrium ferment filtrate includes the following steps: S1 inoculation culture: the activated strain in logarithmic growth phase is inoculated into the seed tank which is sterilized in advance and into which nitrogen gas is introduced to replace air, and incubated at constant temperature for 18h, until the viable cell count of the seed liquid ≥10 9 CFU / mL, OD 600 value reaches 1.2; S2 fermentation: the seed liquid in step S1 is inoculated into the main fermentation tank (1000L) at an inoculation amount of 3%, and the carbon dioxide concentration in the fermentation tank is maintained at 5%, and the fermentation is controlled by stages: 1) The pH value is controlled at 6.0-6.5, the glucose concentration is 1%, the temperature is 37±1℃, and the cultivation is carried out for 24h in the growth stage; 2) In the active substance synthesis stage, the temperature is reduced to 35℃, the pH is adjusted to 5.8-6.0, and the glucose concentration is limited to 0.5%, so as to induce the synthesis of active substances, including polysaccharides and polypeptides, by the bacteria, and the fermentation period is 60h; S3 cell lysis and filtrate separation: 0.1% lysozyme is added, treated at 37℃ for 2h, and then separated by centrifugation and microfiltration in sequence to obtain the Schizochytrium fermentum filtrate.

[0033] Example 2 A condensed firming essence spray, the mass percentage composition of which is as follows: Schizochytrium fermentum filtrate 83.56%, anti-wrinkle agent A 6.5%, anti-wrinkle agent B 3.5%, preservative 2.5%, nicotinamide 1.5%, anti-wrinkle agent C 1%, winter essence 1%, glycyrrhizic acid dipotassium 0.11%, allantoin 0.11%, fuller sugar 0.11%, hydrolyzed collagen 0.11%.

[0034] The mass parts of anti-wrinkle agent A are as follows: Water 60 parts Butanediol 1 part Trehalose 0.1 part 1,2-hexanediol 1 part Ethylhexylglycerin 0.1 part Arginine / lysine polypeptide 0.1 part.

[0035] The mass parts of anti-wrinkle agent B are as follows: Water 60 parts Glycerin 1 part Octanediol 0.1 part Acetyl hexapeptide-8 0.01 part Ethylhexylglycerin 0.01 part.

[0036] The preservative includes 1,2-pentanediol and p-hydroxyacetophenone, and the mass ratio of 1,2-pentanediol to p-hydroxyacetophenone is 2:1.

[0037] The mass parts of winter essence are as follows: Schizochytrium fermentum filtrate 85 parts Glycerin 2 parts 1,2-pentanediol 1 part Radix ophiopogonis extract 2 parts Red ganoderma extract 0.5 part P-hydroxyacetophenone 0.05 part.

[0038] The fullerenes sugar comprises zinc gluconate and fullerene, and the mass ratio is 2:1.

[0039] The preparation method of the Schizosaccharomyces ferment filtrate comprises the following steps: S1 inoculation culture: the activated strain in logarithmic growth phase is inoculated into the seed tank in advance sterilized and with nitrogen gas replacing air at an inoculation amount of 5% (volume ratio), and constant temperature culture is carried out for 18 h, until the viable cell count of the seed liquid is ≥10 9 CFU / mL, OD 600 value reaches 1.2; S2 fermentation: the seed liquid in step S1 is inoculated into the main fermentation tank 1000L at an inoculation amount of 3%, the carbon dioxide concentration in the fermentation tank is maintained at 5%, and the following segmented culture control is adopted: 1) the growth stage controls the pH value to be 6.0-6.5, the glucose concentration is 1%, and the temperature is 37±1℃, and the cultivation is carried out for 24 h; 2) the active substance synthesis stage: the temperature is reduced to 35℃, the pH is adjusted to 5.8-6.0, the glucose concentration is limited to 0.5%, the bacteria are induced to synthesize active substances including polysaccharides and polypeptides, and the fermentation period is 60 h; S3 cell lysis and filtrate separation: 0.1% lysozyme is added, 37℃ treatment is carried out for 2 hours, and then centrifugal separation and microfiltration are carried out in sequence to obtain the Schizosaccharomyces ferment filtrate.

[0040] The preparation method further comprises desalination and concentration, and the specific steps are as follows: the filtrate in step S3 is desalted by nanofiltration, the nanofiltration membrane has a molecular weight cut-off of 300-500 Da, the active small molecules are retained, and the salt is removed; then low-temperature vacuum concentration is adopted, the temperature is controlled to be ≤40℃, the vacuum degree is-0.08 to-0.09 MPa, and the filtrate is concentrated to 1 / 5 of the original volume.

[0041] Example 3 A condensed firming essence spray, which comprises the following components in mass percentage: Schizosaccharomyces ferment filtrate 83.56%, anti-wrinkle agent A 6.5%, anti-wrinkle agent B 3.5%, preservative 2.5%, nicotinamide 1.5%, anti-wrinkle agent C 1%, winter essence 1%, di-potassium glycyrrhizinate 0.11%, allantoin 0.11%, fullerenes sugar 0.11%, and hydrolyzed collagen 0.11%.

[0042] The mass parts of anti-wrinkle agent A are as follows: Water 70 parts Butylene glycol 1.5 parts Trehalose 0.3 parts 1,2-hexanediol 1.5 parts Ethylhexylglycerin 0.2 parts Arginine / lysine polypeptide 0.3 parts.

[0043] The mass parts composition of anti-wrinkle agent B is as follows: Water 72 parts Glycerin 2 parts Octanediol 0.5 parts Acetyl hexapeptide-8 0.03 parts Ethylhexylglycerin 0.08 parts.

[0044] The preservative includes 1,2-pentanediol and p-hydroxyacetophenone, and the mass ratio of 1,2-pentanediol to p-hydroxyacetophenone is 2.5:1.

[0045] The mass parts composition of Dongling enzyme is as follows: Schizosaccharomyces fermentate filtrate 90 parts Glycerin 6 parts 1,2-pentanediol 3 parts Radix ophiopogonis extract 3 parts Red Ganoderma extract 1.2 parts P-hydroxyacetophenone 0.16 parts.

[0046] Fuller sugar includes zinc gluconate and fullerene, and the mass ratio is 3:1.

[0047] The preparation method of the Schizosaccharomyces fermentate filtrate includes the following steps: S1 inoculation culture: inoculate the activated strain in the logarithmic growth phase into the seed tank which is sterilized in advance and into which nitrogen is introduced to replace air at an inoculation amount of 8% (volume ratio), and incubate at constant temperature for 20h, so that the viable cell count of the seed liquid is ≥10 9C FU / mL, OD 600 value reaches 1.3; S2 fermentation: inoculate the seed liquid in step S1 into a main fermentation tank 1000L at an inoculation amount of 4%, maintain the carbon dioxide concentration in the fermentation tank at 6%, and use stepwise culture control: 1) Control the pH value to be 6.0-6.5, the glucose concentration to be 1.5%, and the temperature to be 37±1℃ in the growth stage, and cultivate for 24h; 2) Active substance synthesis stage: reduce the temperature to 35℃, adjust the pH to 5.8-6.0, and limit the glucose concentration to 0.8% to induce the synthesis of active substances, including polysaccharides and polypeptides, by the bacterial cells, and the fermentation period is 65h; S3 bacterial lysis and filtrate separation: add 0.15% lysozyme, treat at 37℃ for 1.5h, and then sequentially pass through centrifugal separation and microfiltration to obtain the Schizosaccharomyces fermentate filtrate.

[0048] The preparation method further comprises desalination and concentration, and the specific steps are as follows: the filtrate in step S3 is desalted by nanofiltration, the nanofiltration membrane has a molecular weight cut-off of 300-500 Da, the active small molecules are retained, and the salts are removed; then low-temperature vacuum concentration is used again, the temperature is controlled to be less than or equal to 40 DEG C, the vacuum degree is-0.08 to-0.09 MPa, and the filtrate is concentrated to 1 / 8 of the original volume.

[0049] Example 4 A condensed firming essence spray has the following mass percentage composition: Schizochytrium limulus fermentation product filtrate 83.56%, anti-wrinkle agent A 6.5%, anti-wrinkle agent B 3.5%, preservative 2.5%, nicotinamide 1.5%, anti-wrinkle agent C 1%, winter essence 1%, glycyrrhizic acid dipotassium 0.11%, allantoin 0.11%, fuller sugar 0.11%, hydrolyzed collagen 0.11%.

[0050] The mass parts of anti-wrinkle agent A are as follows: Water 75 parts Butanediol 1.8 parts Trehalose 0.4 parts 1,2-hexanediol 1.8 parts Ethylhexylglycerin 0.25 parts Arginine / lysine polypeptide 0.45 parts.

[0051] The mass parts of anti-wrinkle agent B are as follows: Water 82 parts Glycerin 2.8 parts Octanediol 0.7 parts Acetyl hexapeptide-8 0.04 parts Ethylhexylglycerin 0.13 parts.

[0052] The preservative comprises 1,2-pentanediol and p-hydroxyacetophenone, and the mass ratio of 1,2-pentanediol to p-hydroxyacetophenone is 2.5:1.

[0053] The mass parts of winter essence are as follows: Schizochytrium limulus fermentation product filtrate 90 parts Glycerin 8 parts 1,2-pentanediol 4.5 parts Radix ophiopogonis extract 3.2 parts Red ganoderma extract 1.8 parts P-hydroxyacetophenone 0.28 parts.

[0054] The fuller sugar comprises zinc gluconate and fullerene, and the mass ratio is 5:1.

[0055] The preparation method of the Schizochytrium limulus fermentation product filtrate comprises the following steps: S1 inoculation culture: the activated strain in logarithmic growth phase is inoculated into the seed tank in advance sterilized and replaced with nitrogen gas at an inoculation amount of 5%-10% (volume ratio), and incubated at constant temperature for 24 h, until the viable cell count of the seed liquid is ≥10 9 CFU / mL, OD 600 value reaches 1.4; S2 fermentation: the seed liquid in step S1 is inoculated into the main fermentation tank 1000L at an inoculation amount of 4.8-5%, and the carbon dioxide concentration in the fermentation tank is maintained at 7.2%, and the cultivation is controlled in stages: 1) the pH value is controlled at 6.0-6.5, the glucose concentration is 1.8%, the temperature is 37±1℃, and the cultivation is carried out for 24 h; 2) active substance synthesis stage: the temperature is reduced to 35℃, the pH is adjusted to 5.8-6.0, the glucose concentration is limited to 0.8%, and the strain is induced to synthesize active substances, including polysaccharides and polypeptides, and the fermentation period is 72 h; S3 cell lysis and filtrate separation: 0.18% lysozyme is added, treated at 37℃ for 1.8 h, then separated by centrifugation and microfiltration in turn to obtain the Schizochytrium fermentum filtrate.

[0056] The preparation method further comprises desalting and concentration, and the specific steps are as follows: the filtrate in step S3 is desalted by nanofiltration, the nanofiltration membrane has a molecular weight cut-off of 300-500 Da, retains active small molecules, and removes salts; then low-temperature vacuum concentration is used, the temperature is controlled at ≤40℃, the vacuum degree is -0.08 to -0.09 MPa, and the filtrate is concentrated to 1 / 5 of the original volume.

[0057] Example 5 A condensed firming essence spray, the mass percentage composition of which is as follows: Schizochytrium fermentum filtrate 83.56%, anti-wrinkle agent A 6.5%, anti-wrinkle agent B 3.5%, preservative 2.5%, nicotinamide 1.5%, anti-wrinkle agent C 1%, winter essence 1%, di-potassium glycyrrhizinate 0.11%, allantoin 0.11%, fuller sugar 0.11%, and hydrolyzed collagen 0.11%.

[0058] The mass parts of anti-wrinkle agent A are as follows: Water 80 parts Butanediol 2 parts Trehalose 0.5 parts 1,2-hexanediol 2 parts Ethylhexylglycerin 0.3 parts Arginine / lysine polypeptide 0.5 parts.

[0059] The mass parts of anti-wrinkle agent B are as follows: Water 84 parts Glycerol 3 parts Octanediol 0.8 parts Acetyl hexapeptide-80.05 parts Ethylhexylglycerin 0.15 parts.

[0060] The preservative includes 1,2-pentanediol and p-hydroxyacetophenone, and the mass ratio of 1,2-pentanediol to p-hydroxyacetophenone is 3:1.

[0061] The mass parts of Dongling enzyme are as follows: Schizosaccharomyces ferment filtrate 95 parts Glycerol 10 parts 1,2-pentanediol 5 parts Ophiopogon root extract 3.5 parts Red Ganoderma extract 2 parts P-hydroxyacetophenone 0.3 parts.

[0062] Fuller sugar includes zinc gluconate and fullerene, and the mass ratio is 6:1.

[0063] The preparation method of the Schizosaccharomyces ferment filtrate includes the following steps: S1 inoculation culture: the activated strain in the logarithmic growth phase is inoculated into the seed tank which is sterilized in advance and into which nitrogen is introduced to replace air, and is cultured at constant temperature for 18-24h, until the viable cell count of the seed liquid is ≥10 9 CFU / mL, OD 600 value reaches 1.2-1.5; S2 fermentation: the seed liquid in step S1 is inoculated into the main fermentation tank (1000-5000L) at an inoculation amount of 3%-5%, the carbon dioxide concentration in the fermentation tank is maintained at 5%-8%, and the following segmented culture control is adopted: 1) The pH value is controlled at 6.0-6.5, the glucose concentration is 1%-2%, and the temperature is 37±1℃, and the cultivation is carried out for 24h in the growth stage; 2) In the active substance synthesis stage, the temperature is reduced to 35℃, the pH is adjusted to 5.8-6.0, and the glucose concentration is limited to 0.5%-1%, so as to induce the synthesis of active substances, including polysaccharides and polypeptides, by the bacteria, and the fermentation period is 60-72h; S3 cell lysis and filtrate separation: 0.1%-0.2% lysozyme is added, treated at 37℃ for 1-2h, and then separated by centrifugation and microfiltration in sequence to obtain the Schizosaccharomyces ferment filtrate.

[0064] The preparation method further includes desalination and concentration, and the specific steps are as follows: the filtrate in step S3 is desalted by nanofiltration, the nanofiltration membrane has a molecular weight cut-off of 300-500Da, the active small molecules are retained, and the salt is removed; then low-temperature vacuum concentration is adopted, the temperature is controlled to be ≤40℃, the vacuum degree is-0.08 to-0.09MPa, and the filtrate is concentrated to 1 / 5-1 / 10 of the original volume.

[0065] A clinical evaluation model of randomized, double-blind, self- left and right face control was used to test the effect of the firming serum spray prepared in Example 3.

[0066] I. Clinical experiment (n = 42, Asian women aged 35-55, Fitzpatrick type III-IV, 50% sensitive skin) Placebo: 83.56% deionized water + preservative + glycol system, appearance, pH, and viscosity consistent with the spray.

[0067] Positive control: 3% bovine serum solution.

[0068] Primary endpoints (D0, D7, D14, D28) II. In vitro experiment (3D recombined human epidermis + fibroblast co-culture model) 2.1 Synergistic antioxidant effect of double yeast Conclusion: Synergy index CI = 1.31 (> 1.1 is synergistic).

[0069] 2.2 Anti-wrinkle agents A + B nerve block 2.3 Melanin pathway 2.4 Collagen neogenesis III. Microecological safety (16S rDNA sequencing, n = 10) Conclusion: The product does not destroy the diversity of skin flora and significantly inhibits potential pathogenic bacteria.

[0070] IV. Stability and activity retention 40°C / 75%RH accelerated for 90 days: Acetyl hexapeptide-8 retention rate 96.5% (UPLC) Arginine / lysine polypeptide retention rate 97.2% pH, appearance, and microorganisms meet the "Cosmetic Safety Technology Specifications" 2022.

[0071] It should be pointed out finally that the above embodiments are only used to illustrate the technical solutions of the present application, but not to limit the same; and although the present application has been described in detail with reference to the foregoing embodiments, it should be appreciated by those skilled in the art that the technical solutions recorded in the foregoing embodiments can be modified, or some technical features thereof can be replaced equivalently; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. A time-condensed firming serum spray, characterized in that: The mass percentage composition is as follows: Schizosaccharomyces pombe fermentation product filtrate 83.56%, anti-wrinkle agent A 6.5%, anti-wrinkle agent B 3.5%, preservative 2.5%, nicotinamide 1.5%, anti-wrinkle agent C 1%, winter essence 1%, dipotassium glycyrrhizinate 0.11%, allantoin 0.11%, fuller sugar 0.11%, hydrolyzed collagen 0.11%, and water 0.11%.

2. The serum-in-water emulsion according to claim 1, wherein: The preparation method of the Schizosaccharomyces pombe fermentation product filtrate comprises the following steps: S1 inoculation culture: the activated strain in logarithmic growth phase is inoculated into the seed tank which is sterilized in advance and replaced with nitrogen instead of air at 5-10% inoculation amount (volume ratio), and incubated at constant temperature for 18-24h, until the viable cell count of the seed liquid is ≥10 9 CFU / mL, OD 600 value reaches 1.2-1.5; S2 fermentation: inoculate the seed liquid in step S1 into a main fermentation tank (1000-5000L) at an inoculation amount of 3%-5%, maintain the carbon dioxide concentration in the fermentation tank at 5%-8%, and use stepwise culture control: 1) In the growth stage, control the pH value to be 6.0-6.5, the glucose concentration to be 1%-2%, and the temperature to be 37±1℃, and cultivate for 24h; 2) In the active substance synthesis stage, reduce the temperature to 35℃, adjust the pH to 5.8-6.0, limit the glucose concentration to 0.5%-1%, and induce the bacteria to synthesize active substances, including polysaccharides and polypeptides, and the fermentation cycle is 60-72h; S3 cell lysis and filtrate separation: add 0.1%-0.2% lysozyme, treat at 37℃ for 1-2h, and then sequentially perform centrifugal separation and microfiltration to obtain the Schizosaccharomyces pombe fermentation product filtrate.

3. The serum-in-hydrogel according to claim 2, wherein: The preparation method further comprises desalination and concentration, and the specific steps are as follows: desalinate the filtrate in step S3 by nanofiltration, the nanofiltration membrane has a molecular weight cut-off of 300-500Da, retains active small molecules, and removes salt; then, perform low-temperature vacuum concentration, control the temperature to be ≤40℃, the vacuum degree to be -0.08 to -0.09MPa, and concentrate the filtrate to 1 / 5-1 / 10 of the original volume.

4. The serum-in-hydrogel according to claim 1, wherein: The mass parts of the anti-wrinkle agent A are as follows: Water 60-80 parts Butanediol 1-2 parts Trehalose 0.1-0.5 parts 1,2-hexanediol 1-2 parts Ethylhexyl glycerin 0.1-0.3 parts Arginine / lysine polypeptide 0.1-0.5 parts.

5. The serum according to claim 1, wherein the serum is a serum-in-hydrogel. The mass parts of the anti-wrinkle agent B are as follows: Water 60-84 parts Glycerin 1-3 parts Octanediol 0.1-0.8 parts Acetyl hexapeptide-8 0.01-0.05 parts Ethylhexyl glycerin 0.01-0.15 parts.

6. The serum-in-hydrogel according to claim 1, wherein: The preservative comprises 1,2-pentanediol and p-hydroxyacetophenone, and the mass ratio of 1,2-pentanediol to p-hydroxyacetophenone is (2-3):

1.

7. The serum-in-hydrogel according to claim 1, wherein: The mass parts of the winter essence are as follows: Schizosaccharomyces pombe fermentation product filtrate 85-95 parts Glycerin 2-10 parts 1,2-pentanediol 1-5 parts Radix ophiopogonis extract 2-3.5 parts Red ganoderma extract 0.5-2 parts P-hydroxyacetophenone 0.05-0.3 parts.

8. The serum-in-hydrogel according to claim 1, wherein: The fuller sugar comprises zinc gluconate and fullerene, and the mass ratio is (1-6):1.

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