Preparation process of high-activity fermented food-grade snail stock solution and special moisturizing and repairing mask for pre-pregnancy and pregnancy

Abstract

The application discloses a high-activity fermentation type food-grade snail stock solution preparation process and a special moisturizing and repairing mask for pre-pregnancy and pregnancy, and belongs to the technical field of biological skin care raw material preparation. The application adopts a composite food-grade probiotic low-temperature segmented directional fermentation process: white jade snail mucus is collected in a sterile environment at 22-25 DEG C, a crude stock solution is mixed with plant lactobacillus and bifidobacterium compound bacterial liquid at 92:8, and three-stage temperature control treatment is carried out through 4-8 DEG C pre-fermentation, 22-26 DEG C directional zymolysis and 0-4 DEG C ripening, and then 0-5 DEG C ultrafiltration sterilization is carried out, so that a food-grade refined stock solution with an activity retention of greater than or equal to 95% and natural antibacterial capacity is obtained, and no chemical is added in the whole process. The stock solution is used to develop double-specification masks: the pre-pregnancy mask contains 75-85 wt% of the stock solution, is matched with sodium hyaluronate and glycerol, and focuses on repairing and improving dull and dry wrinkles. The application solves the short board of the traditional process, accurately adapts to fragile skin in special periods, can be mass-produced, and has a wide application prospect.

Description

Technical Field

[0001] This invention relates to the field of natural biological skin care raw material processing technology, and in particular to a preparation process of highly active fermented food-grade snail extract and a moisturizing and repairing face mask specially designed for preconception and pregnancy. Background Technology

[0002] In the current field of natural skincare ingredient processing, the resource utilization of snail mucus has reached a certain industrial scale. However, existing processes and end products face multiple insurmountable technical bottlenecks, making it impossible to meet the skincare needs of special groups such as those preparing for pregnancy and during pregnancy. Specific problems are as follows: 1. Extremely low active ingredient retention rate: Commercially available snail mucin generally uses physical pressing and room temperature extraction processes. The heat-sensitive and mechanically shear-sensitive functional components in snail mucin, such as glycoproteins, antibacterial peptides, allantoin, natural polysaccharides, and active antioxidant enzymes, are severely damaged by heat and mechanically sheared during processing. Tests show that the active nutrient retention rate is only 50% to 65%, and the utilization rate of raw material efficacy is less than two-thirds, which fails to achieve the expected skin care effect.

[0003] The risk of allergic reactions remains high: Unmodified snail mucus contains a large number of foreign macromolecular proteins. These substances are exogenous antigens to human skin and can easily trigger an immune response in sensitive skin. Clinical feedback shows that more than 32% of users experienced redness, itching, and papules after using regular snail products. The risk is even higher for people whose skin barrier is damaged and whose tolerance is reduced due to hormonal fluctuations during pregnancy.

[0004] Insufficient safety of preservative systems: To ensure the shelf life of snail extract and end products, the industry commonly adds chemical preservatives such as parabens and phenoxyethanol. Some products even use polyol compound preservative systems. At the same time, in order to highlight efficacy, highly irritating ingredients such as retinol, fruit acids, and salicylic acid are often added. None of the above substances are on the list of safe ingredients during pregnancy, and there are potential health risks for women who are trying to conceive or pregnant.

[0005] Lack of target audience suitability: Existing snail-based skincare products lack segmented design for different target groups. They do not consider the differences between the needs of those trying to conceive, who require improvement in dullness and early signs of aging, and the needs of those pregnant, who require minimalist, low-sensitivity moisturizing. The uniform formulas cannot adapt to the changes in skin condition at different stages, resulting in a lack of suitable products for the target audience.

[0006] In summary, there is a lack of existing technologies that can simultaneously achieve high activity retention, removal of allergenic proteins, natural self-antibacterial properties, no chemical additives, and precise suitability for pregnant women and women in the pregnancy period. The relevant technological gaps urgently need to be filled. Summary of the Invention

[0007] In view of this, the present invention provides a process for preparing highly active fermented food-grade snail extract and a moisturizing and repairing face mask specifically for those preparing for pregnancy and during pregnancy, in order to solve or alleviate one of the technical problems existing in the prior art, and at least provide a beneficial alternative.

[0008] The technical solution of this invention is implemented as follows: a process for preparing a highly active fermented food-grade snail extract, comprising the following steps: (1) Aseptic collection of mucus from live snails: In a sterile environment at a constant temperature of 22~25℃, the mucus from live white snails is naturally induced by physical gentle induction. The collected mucus is immediately sealed and collected at low temperature, and then filtered with a 200-mesh sterile filter to remove mud, sand, tissue debris and impurities to obtain crude snail stock solution. (2) Preparation of compound fermentation broth: Food-grade Lactobacillus plantarum and Bifidobacterium were selected as the fermentation strains and prepared with sterile pure water as the solvent to obtain a viable bacteria content of 1×10⁻⁶. 7 ~1×10 8 Fermentation broth with CFU / mL; (3) Three-stage gradient closed temperature-controlled fermentation: The crude snail stock solution and the fermentation liquid were mixed evenly at a volume ratio of 92:8 and fermented in a closed, light-proof, and sterile environment throughout the process. The fermentation process went through three stages in sequence: low temperature pre-fermentation, medium temperature directional fermentation, and ultra-low temperature live-locking maturation. The temperature of the low temperature pre-fermentation stage was controlled at 4~8℃ and the static culture was carried out for 18~24h; the temperature of the medium temperature directional fermentation stage was controlled at 22~26℃ and the fermentation was carried out for 48~72h; the temperature of the ultra-low temperature live-locking maturation stage was controlled at 0~4℃ and the maturation was carried out for 12~16h. (4) Low-temperature precision filtration sterilization: Ultrafiltration is used at a low temperature of 0~5℃ to remove bacterial residues and residual impurities, resulting in food-grade refined snail extract with an activity retention rate of ≥95% and natural antibacterial properties.

[0009] Furthermore, the compound fermentation liquid in step (2) is made by mixing Lactobacillus plantarum and Bifidobacterium at a ratio of 3:2 based on the dry weight of the bacteria. No chemical preservatives, acid-base extraction aids, or organic solvents are added during the entire fermentation process.

[0010] Furthermore, the preferred parameters for the low-temperature pre-fermentation stage in step (3) are 6°C and 22h; the preferred parameters for the mesophilic directional fermentation stage are 24°C and 62h; and the preferred parameters for the ultra-low temperature activation maturation stage are 2°C and 15h.

[0011] Furthermore, the ultrafiltration process in step (4) uses a hollow fiber membrane with a molecular weight cutoff of 10 kDa and controls the transmembrane pressure to be 0.1~0.2 MPa. The resulting refined snail stock solution has an antibacterial rate of ≥99% against Staphylococcus aureus and an antibacterial rate of ≥98% against Candida albicans.

[0012] Furthermore, the content of allergenic macromolecular foreign proteins in the obtained refined snail stock solution is ≤0.15mg / g, and the oral acute toxicity test reaches the practically non-toxic level, which meets the food contact safety standard.

[0013] Furthermore, the highly active fermented food-grade snail extract prepared by the process described in any one of claims 1 to 5 is used in a face mask with two independent formulas: a formula for preparing for pregnancy and a simplified formula for pregnancy.

[0014] Furthermore, the formula for preparing for pregnancy contains the following components by weight percentage: 75-85 wt% refined fermented snail extract, 1-3 wt% food-grade sodium hyaluronate, 2-4 wt% vegetable glycerin, and the balance being sterile deionized water. No additional preservatives, fragrances, or irritating additives are added to the formula.

[0015] Furthermore, the simplified pregnancy formula contains the following components by weight percentage: 60-70 wt% refined fermented snail extract, 1-2 wt% medical-grade small molecule moisturizing dextran, 1-3 wt% vegetable glycerin, and the balance being sterile deionized water. All unnecessary functional additives have been removed from the formula.

[0016] Furthermore, the mask base material is made of natural cottonseed sheet, and the product relies on the natural antibacterial properties of snail extract to achieve shelf-life stability without the need for additional preservatives. The finished product has been verified to be non-irritating through skin irritation tests, with a sensitization rate of 0%.

[0017] The embodiments of the present invention have the following advantages due to the adoption of the above technical solutions: 1. Breakthrough improvement in active ingredient retention: Compared to the traditional physical extraction process with an active ingredient retention rate of only 50%~65%, this process, through low-temperature segmented fermentation and low-temperature treatment throughout the process, stably increases the retention rate of active ingredients to ≥95%. All the functional components in snail mucus, such as natural glycoproteins, antibacterial peptides, allantoin, natural polysaccharides, and active antioxidant enzymes, are completely preserved. Cell experiments have verified that its repair effect in promoting the migration of keratinocytes is 2.8 times that of traditional essences, and the moisturizing time is extended to more than 72 hours.

[0018] Safety is upgraded in all dimensions: On the one hand, more than 90% of allergenic macromolecular foreign proteins are removed through probiotic-directed fermentation. Patch tests have verified that the allergy rate has been reduced from 32% in traditional products to 0.8%. On the other hand, the organic acids, bacteriocins and other endogenous metabolites produced during the fermentation process endow the original liquid with natural self-antibacterial ability. No chemical preservatives need to be added. The original liquid meets the food contact safety standard in oral toxicity tests, and the antibacterial rates against Staphylococcus aureus and Candida albicans reach 99.2% and 98.7% respectively.

[0019] Precise segmentation of target groups: The dual-formula design perfectly matches the differentiated needs of the preconception period and pregnancy. The preconception version combines high activity and multiple functions to solve problems such as dull skin and fine lines before pregnancy; the pregnancy version uses a minimalist formula, retaining only basic moisturizing and repairing ingredients and avoiding all risky substances during pregnancy. It has been tested on 300 pregnant volunteers and no irritation reactions have occurred.

[0020] High feasibility for industrialization: The process parameters are stable and controllable, all raw materials are food-grade compliant materials, there are no high temperature, high pressure and highly toxic reagents used, it can be directly connected to existing cosmetic production lines to achieve large-scale mass production, the unit raw material cost is 42% lower than that of imported similar high-activity snail extract, and it has broad market application prospects.

[0021] The above overview is for illustrative purposes only and is not intended to be limiting in any way. In addition to the illustrative aspects, embodiments, and features described above, further aspects, embodiments, and features of the invention will become readily apparent from the accompanying drawings and the following detailed description. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0023] Figure 1 This is a flow chart of the preparation process of the highly active fermented food-grade snail stock solution of the present invention; Figure 2 This is a flowchart illustrating the preparation process of the pre-pregnancy moisturizing and repairing facial mask of the present invention. Figure 3 This is a flowchart illustrating the preparation process of the simplified moisturizing and repairing facial mask for pregnancy according to the present invention. Figure 4 This is a detailed flowchart of the three-stage gradient fermentation process of the present invention. Detailed Implementation

[0024] In the following description, only certain exemplary embodiments are briefly described. As those skilled in the art will recognize, the described embodiments can be modified in various ways without departing from the spirit or scope of the invention. Therefore, the drawings and description are considered to be exemplary in nature and not restrictive.

[0025] The embodiments of the present invention will now be described in detail with reference to the accompanying drawings.

[0026] Example 1: Preparation of Highly Active Fermented Food-Grade Snail Stock Solution Fresh white jade snails Shell height 3-4cm, healthy and free from disease Shandong snail breeding base Lactobacillus plantarum Food grade, strain number CGMCC No. 18234 China General Microbiological Culture Collection Center Bifidobacteria Food grade, strain number CGMCC No. 17562 China General Microbiological Culture Collection Center sterile deionized water Conductivity ≤10μS / cm Lab-made sterile cleanroom Cleanliness level ISO 7 Own workshop Hollow fiber ultrafiltration system 10kDa molecular weight cutoff Tianjin Membrane Technology Co., Ltd. 2. Preparation steps (1) Aseptic collection of mucus from live snails: The white jade snails were transferred to a 22°C constant temperature aseptic clean room and fasted for 12 hours. The mucus was then physically induced by a soft brush and collected immediately in a sterile glass container. The mucus was then coarsely filtered through a 200-mesh sterile stainless steel filter to remove mud, sand and snail tissue debris, resulting in 920 kg of crude snail stock solution. The pH of the stock solution after coarse filtration was 6.8 and the viscosity was 120 mPa·s.

[0027] (2) Preparation of compound fermentation broth: Weigh Lactobacillus plantarum and Bifidobacterium freeze-dried powder at a ratio of 3:2 by dry weight of the bacteria, add sterile deionized water to reconstitute, shake to mix well, and then adjust the viable bacteria concentration to 5×10 using a hemocytometer. 7 A total of 100 kg of fermentation broth was prepared using CFU / mL.

[0028] Low-temperature pre-fermentation 6℃ 22h <![CDATA[The viable cell count increased to 1.2×10 8 CFU / mL, and the pH of the original solution decreased to 6.4]]> Medium-temperature directional fermentation 24℃ 62h The content of macromolecular proteins decreased to 0.12 mg / g, while the content of reducing sugars increased to 2.3 mg / g. Cryogenic Cryogenic Maturation 2℃ 15h <![CDATA[The viable count is stable at 1.1×10 8 CFU / mL, and the pH is stable at 5.8]]> (4) Low-temperature precision filtration and sterilization: The original liquid after fermentation is pumped into a 0~5℃ low-temperature hollow fiber ultrafiltration system, the transmembrane pressure is controlled at 0.15MPa, the molecular weight cutoff is 10kDa, the bacterial residue and macromolecular impurities are removed, and finally 987kg of refined fermented snail original liquid is obtained with a yield of 98.7%.

[0029] Bioactive retention rate Guidelines for Efficacy Evaluation of Cosmetic Raw Materials (QB / T 5668-2021) 95.7% ≥95% Allergenic macromolecular protein content Coomassie Brilliant Blue 0.11mg / g ≤0.15mg / g Staphylococcus aureus inhibition rate GB 15979-2002 99.3% ≥99% Candida albicans inhibition rate GB 15979-2002 98.8% ≥98% Chemical preservatives detected GB / T 39993-2021 Not detected Not detected acute oral toxicity test GB 15193.3-2014 Practically non-toxic Food contact grade Example 2: Moisturizing and Repairing Face Mask for Pregnancy Preparation (Pregnancy Preparation Version) Refined Fermented Snail Extract 80% Preparation in Example 1 self made Food-grade small molecule sodium hyaluronate 2% Molecular weight 8000~10000 Da Bloomage Biotechnology Co., Ltd. Plant-derived glycerin 3% Purity ≥ 99.5% Malaysian Coconut Palm Group sterile deionized water margin Conductivity ≤10μS / cm Lab-made Natural cottonseed membrane cloth Facial patch, 25g / piece 45g / m² Hangzhou Reliable Nursing Products Co., Ltd. 2. Preparation process (1) Heat sterile deionized water to 40°C, add plant-derived glycerin and stir until completely dissolved, cool to 30°C and add food-grade small molecule sodium hyaluronate, stir at low speed for 30 minutes until evenly dispersed; (2) Add the refined fermented snail stock solution prepared in Example 1, stir at 200 r / min for 20 min, mix evenly and let stand for 1 h to defoam; (3) After the mixed liquid is sterilized by filtering through a 0.22μm filter cartridge, it is quantitatively injected into sterile aluminum foil bags, each bag containing 25g of liquid, and at the same time, folded natural cottonseed membrane cloth is placed in it and heat-sealed. (4) The finished products are placed in a constant temperature warehouse at 25℃ for 7 days. After passing the random inspection, they are packed and stored.

[0030] Appearance Visual inspection The liquid was clear and transparent, with no sediment. qualified pH value GB / T 13531.1-2008 5.7 Matches the skin's physiological pH Total bacterial count Cosmetic Safety Technical Specifications (2015 Edition) <10 CFU / g qualified Pathogenic bacteria Cosmetic Safety Technical Specifications (2015 Edition) Not detected qualified Pregnancy risk components Cosmetic Safety Technical Specifications (2015 Edition) Not detected qualified Skin hydration rate (after 2 hours of use) Corneometer CM 825 An increase of 47.2% Superior to similar products on the market 4. Results of human trials skin moisture content 38.2% 56.7% 48.4% Skin radiance 62.3 81.5 30.8% Number of fine lines (around the eyes) Article 4.2 Article 1.8 57.1% Adverse reaction incidence - 0% - Example 3: Moisturizing and Repairing Face Mask for Preparing for Pregnancy and During Pregnancy (Simplified Version for Pregnancy) Refined Fermented Snail Extract 65% Preparation in Example 1 self made Medical small molecule moisturizing glucan 1.5% Molecular weight 5000~7000 Da Yantai Andeli Group Co., Ltd. Vegetable glycerin 2% Purity ≥ 99.5% Malaysian Coconut Palm Group sterile deionized water 31.5% Conductivity ≤10μS / cm Lab-made Natural cottonseed membrane cloth Facial patch, 25g / piece 45g / m² Hangzhou Reliable Nursing Products Co., Ltd. 2. Preparation process (1) Add sterile deionized water, vegetable glycerin and medical small molecule moisturizing dextran to the mixing tank in sequence, and stir at 150 r / min for 40 min at 25℃ until all components are completely dissolved. (2) Add the refined fermented snail stock solution prepared in Example 1, continue stirring for 15 min, mix evenly, and then filter through a 0.22 μm filter cartridge for sterilization; (3) The liquid is filled into aluminum foil bags using an aseptic filling line, 25g per bag, and natural cottonseed film is placed in the bags at the same time. After heat sealing, the bags are sterilized by irradiation (dose 8kGy). (4) Store the finished product in a cool and dry place and put it on the market after passing the inspection.

[0031] Appearance Visual inspection The liquid is colorless, transparent, and free of impurities. qualified pH value GB / T 13531.1-2008 5.6 Matches the skin's physiological pH Irritation (rabbit skin test) Cosmetic Safety Technical Specifications (2015 Edition) No erythema, no edema Non-irritating Sensitization (guinea pig maximization test) Cosmetic Safety Technical Specifications (2015 Edition) The sensitization rate was 0. Non-allergenic Shelf life prediction (accelerated test 40℃ / 75%RH) 6 months Active retention rate: 94.2% Meets 24-month shelf life 4. Results of human trials Incidence of skin redness 0 17 cases (14.2%) P＜0.05 Incidence of skin itching 0 12 cases (10.0%) P＜0.05 Increase in skin hydration 32.7% 18.4% P＜0.01 Subject satisfaction 96.7% 78.3% P＜0.01 The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any person skilled in the art can easily conceive of various variations or substitutions within the technical scope disclosed in the present invention, and these should all be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A process for preparing a highly active fermented food-grade snail extract, characterized in that, Includes the following steps: (1) Aseptic collection of mucus from live snails: In a sterile environment at a constant temperature of 22~25℃, the mucus from live white snails is naturally induced by physical gentle induction. The collected mucus is immediately sealed and collected at low temperature, and then filtered with a 200-mesh sterile filter to remove mud, sand, tissue debris and impurities to obtain crude snail stock solution. (2) Preparation of compound fermentation broth: Food-grade Lactobacillus plantarum and Bifidobacterium were selected as the fermentation strains and prepared with sterile pure water as the solvent to obtain a viable bacteria content of 1×10⁻⁶. 7 ~1×10 8 Fermentation broth with CFU / mL; (3) Three-stage gradient closed temperature-controlled fermentation: The crude snail stock solution and the fermentation liquid were mixed evenly at a volume ratio of 92:8 and fermented in a closed, light-proof, and sterile environment throughout the process. The fermentation process went through three stages in sequence: low temperature pre-fermentation, medium temperature directional fermentation, and ultra-low temperature live-locking maturation. The temperature of the low temperature pre-fermentation stage was controlled at 4~8℃ and the static culture was carried out for 18~24h; the temperature of the medium temperature directional fermentation stage was controlled at 22~26℃ and the fermentation was carried out for 48~72h; the temperature of the ultra-low temperature live-locking maturation stage was controlled at 0~4℃ and the maturation was carried out for 12~16h. (4) Low-temperature precision filtration sterilization: Ultrafiltration is used at a low temperature of 0~5℃ to remove bacterial residues and residual impurities, resulting in food-grade refined snail extract with an activity retention rate of ≥95% and natural antibacterial properties.

2. The preparation process of the highly active fermented food-grade snail stock solution according to claim 1, characterized in that, The compound fermentation liquid in step (2) is made by mixing Lactobacillus plantarum and Bifidobacterium at a ratio of 3:2 based on the dry weight of the bacteria. No chemical preservatives, acid-base extraction aids, or organic solvents are added during the entire fermentation process.

3. The preparation process of the highly active fermented food-grade snail stock solution according to claim 1, characterized in that, The preferred parameters for the low-temperature pre-fermentation stage in step (3) are 6°C and 22h; the preferred parameters for the mesophilic directional fermentation stage are 24°C and 62h; and the preferred parameters for the ultra-low temperature activation maturation stage are 2°C and 15h.

4. The preparation process of the highly active fermented food-grade snail stock solution according to claim 1, characterized in that, The ultrafiltration process described in step (4) uses a hollow fiber membrane with a molecular weight cutoff of 10 kDa and controls the transmembrane pressure to be 0.1~0.2 MPa. The resulting refined snail stock solution has an antibacterial rate of ≥99% against Staphylococcus aureus and an antibacterial rate of ≥98% against Candida albicans.

5. The preparation process of the highly active fermented food-grade snail stock solution according to claim 1, characterized in that, The refined snail extract contained ≤0.15mg / g of allergenic macromolecular foreign proteins, and the oral acute toxicity test showed that it reached the practically non-toxic level, meeting the food contact safety standard.

6. A moisturizing and repairing facial mask specifically for use during preconception and pregnancy, characterized in that, The product contains highly active fermented food-grade snail extract prepared by the process described in any one of claims 1 to 5. The face mask has two independent formulas: a formula for preparing for pregnancy and a simplified formula for pregnancy.

7. The moisturizing and repairing facial mask specifically for pregnancy preparation according to claim 6, characterized in that, The formula for preparing for pregnancy contains the following components by weight percentage: 75-85 wt% refined fermented snail extract, 1-3 wt% food-grade sodium hyaluronate, 2-4 wt% vegetable glycerin, and the balance being sterile deionized water. No additional preservatives, fragrances, or irritating additives are added to the formula.

8. The moisturizing and repairing facial mask specifically for pregnancy preparation and repair according to claim 6, characterized in that, The simplified formula for pregnancy contains the following components by weight percentage: 60-70 wt% refined fermented snail extract, 1-2 wt% medical-grade small molecule moisturizing dextran, 1-3 wt% vegetable glycerin, and the balance being sterile deionized water. All unnecessary functional additives have been removed from the formula.

9. The moisturizing and repairing facial mask specifically for pregnancy preparation according to claim 6, characterized in that, The mask base material is made of natural cottonseed sheet. The product relies on the natural antibacterial properties of snail extract to achieve shelf-life stability, without the need for additional preservatives. The finished product has been verified to be non-irritating through skin irritation testing, with a sensitization rate of 0%.

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