Preparation process of a targeted skin care type ellobium auris shell active compound

Abstract

This invention discloses a preparation process for a targeted skincare-enhancing white snail active complex, belonging to the field of functional fermented beverage technology. The beverage uses fresh red grapes, rock sugar, a special fermentation agent, and nano-magnesium-based solid hydrogen flakes as main raw materials, and is produced through specific processes such as ultrasonic-ozone combined cleaning, oxygen-controlled fermentation, solid-liquid coupled pressurized hydrogenation, and low-temperature aging. Its core lies in utilizing the in-situ reaction of nano-magnesium-based solid hydrogen flakes with water to generate hydrogen gas, combined with pressurized hydrogen dissolution technology, to ensure a stable dissolved hydrogen concentration of ≥800 ppb in the beverage, while retaining ≥350 mg / L of total polyphenols. This invention constructs a dual antioxidant system of "grape polyphenols + molecular hydrogen," resulting in a product with an alcohol content of 3.0-5.0% vol, combining the rich fruity aroma of musalas with the refreshing taste of modern sparkling water, showing significant application potential in assisting in lowering blood lipids, anti-oxidation, and relieving physical fatigue.

Description

Technical Field

[0001] This invention relates to the field of bioactive ingredient preparation and cosmetic raw material technology, specifically a preparation process for a targeted skincare-type white snail active complex. Background Technology

[0002] Snail-derived active ingredients have become core ingredients in the high-end beauty industry due to their excellent skin repair, moisturizing, and anti-aging effects. Among them, snail enzymes can promote cell metabolism and accelerate the repair of damaged tissues; glycosaminoglycans have powerful water-locking, moisturizing, soothing, and anti-inflammatory effects; and growth factors can stimulate collagen regeneration and reduce fine lines and wrinkles. The three types of ingredients work synergistically to achieve a full-chain skincare effect of "repair-moisturizing-anti-aging".

[0003] However, existing snail active ingredient extraction and application technologies face several bottlenecks: First, traditional extraction processes often employ high-temperature extraction or single enzymatic hydrolysis, leading to the inactivation of a large number of heat-sensitive growth factors and the destruction of glycosaminoglycan structures, preventing the simultaneous and efficient enrichment of the three core components; second, the extracts often contain residual large-molecule allergenic proteins, which can easily trigger allergic reactions in people with sensitive skin, limiting the product's applicability; third, the active ingredients have poor stability and are prone to oxidation and degradation during room temperature storage, resulting in a decrease in the efficacy of beauty products; fourth, most processes are not optimized for the skin feel requirements of beauty products, and the addition of extracts can easily make the product sticky, affecting the user experience.

[0004] While some improved processes for snail extract exist internationally, they mostly focus on extracting single components, failing to achieve synergistic enrichment and stable compatibility of the three core components. For example, a South Korean company used a low-temperature enzymatic hydrolysis process to extract snail enzymes, achieving an extraction rate of 85%, but the loss rate of glycosaminoglycans and growth factors exceeded 30%. A French laboratory used membrane separation technology to purify snail extracts, reducing sensitization, but failing to solve the problem of storage stability of active ingredients. Therefore, developing a process for preparing a white snail active complex that can simultaneously achieve high extraction rate, high purity, high stability, and low sensitization, and is suitable for the application needs of beauty products, has become a key technology that the industry urgently needs to overcome. Summary of the Invention

[0005] The technical problem to be solved by this invention is to overcome the above-mentioned technical defects.

[0006] To address the above problems, the technical solution of this invention is: a preparation process for a targeted skincare-enhancing white jade snail active complex, comprising the following steps: Step S1, Raw Material Pretreatment and Low-Temperature Fractional Extraction: Select healthy, live white jade snails weighing 60-80g and place them in an environment with a temperature of 20-22℃ and a relative humidity of 65%-70% for 2-3 days without food. Aseptically separate the soft tissue and foot gland secretions, and mix them at a mass ratio of soft tissue:foot gland secretions:sterile ice water = 3:1:10. Adjust the pH of the system to 6.5-7.0. First, extract the snails by stirring at low speed at 4℃ for 2 hours to extract glycosaminoglycans and growth factors. Then, raise the temperature to 15℃ and continue stirring for 1 hour to extract snail enzymes. Combine the two extracts to obtain a mixed extract. Step S2, Ultrasonic-Assisted Targeted Hydrolysis with Compound Enzyme: Add a compound enzyme preparation to the mixed extract. The compound enzyme preparation consists of papain and hyaluronidase in a mass ratio of 2:1, with a total enzyme activity ≥1000 U / mg. The amount of enzyme added is 1.2%–1.8% of the total weight of the raw materials. Adjust the pH of the system to 6.0–6.5, raise the temperature to 45°C, and apply 20kHz ultrasound-assisted treatment at a power of 300W, working for 5 seconds and then intermittently for 3 seconds, for isothermal hydrolysis for 3.5–4 hours. After hydrolysis, inactivate the enzyme in a 75°C water bath for 8 minutes to obtain the hydrolysate. Step S3, Membrane separation and purification: The enzymatic hydrolysate is centrifuged at 4℃ and 8000rpm for 30min. The supernatant is then sequentially purified by passing it through a 5000Da ultrafiltration membrane and a 1000Da nanofiltration membrane. The ultrafiltration pressure is 0.25-0.35MPa, the nanofiltration pressure is 0.5-0.6MPa, and the feed temperature is controlled at 4-8℃. The 1000Da nanofiltration permeate is collected to obtain the core active ingredient solution of white jade snail. Step S4, Steady-state compounding of active ingredients: Add compounding adjuvants to the core active ingredient solution. The compounding adjuvants are composed of glycerin, 1,3-propanediol and panthenol in a mass ratio of 5:3:2. The amount of compounding adjuvants added is 10% to 15% of the weight of the active ingredient solution. Stir and mix at low speed for 30 minutes at 25°C to obtain a targeted skin care type white snail active complex. Step S5, Aseptic filling and storage: After aseptic filtration of the complex through a 0.22μm microporous membrane, it is vacuum-filled in the dark and stored at 4–8℃.

[0007] Furthermore, in step S1, the white jade snails should be selected from adult snails with intact shells and clear crawling tracks. During the fasting and quiet recuperation period, they should be rinsed once a day with sterile water at 25°C to remove feces and impurities.

[0008] Furthermore, in step S2, the power density of the ultrasonic co-processing is 0.55 W / cm², and the enzyme is stirred once every 30 minutes during the enzymatic hydrolysis process at a stirring rate of 50 r / min for 5 minutes.

[0009] Furthermore, the 5000Da ultrafiltration membrane mentioned in step S3 is used to retain large molecular proteins, and the permeate from the 1000Da nanofiltration membrane contains snail enzyme content ≥18mg / g, glycosaminoglycan content ≥25mg / g, and growth factor content ≥12mg / mL.

[0010] Furthermore, the compounding agent mentioned in step S4 can be replaced with a mixed system of glycerol, xylitol, and vitamin B5 in a mass ratio of 4:4:2. After compounding, the stability of the active ingredients at room temperature is improved by more than 40%.

[0011] Furthermore, the process achieves a comprehensive extraction rate of ≥95.3% for the three core components: snail enzyme, glycosaminoglycan, and growth factors.

[0012] Furthermore, the total purity of the snail enzyme, glycosaminoglycan, and growth factors in the complex is ≥99.1%, the retention rate of active ingredients is ≥92.7% after 12 months of storage at room temperature, the skin fibroblast proliferation promotion rate is ≥52.6%, and there are no sensitizing impurities remaining.

[0013] Furthermore, the composite exhibits optimal stability in the pH range of 5.5–6.5 and a conductivity of 200–400 μS / cm.

[0014] Furthermore, the complex is added to serums, face creams, eye creams, and body lotions at a mass percentage of 5% to 15% for skin barrier repair, fine line reduction, and sensitive skin soothing.

[0015] Furthermore, when applied to face cream, the addition amount is 10%, and after 28 days of use, the skin barrier repair rate is ≥48.5%, and the dry lines are reduced by ≥32.7%.

[0016] The advantages of this invention compared to existing technologies are: (1) Synergistic enrichment of core components: The combination of low-temperature fractional extraction and ultrasonic synergistic enzymatic hydrolysis enables the simultaneous and efficient extraction of three types of components: snail enzyme, glycosaminoglycan, and growth factors. The comprehensive extraction rate is over 95.3%, which is much higher than the 70% to 80% of traditional processes. (2) High purity and low allergenicity: The graded membrane separation technology completely removes allergenic proteins and impurities, and the total purity of active ingredients is ≥99.1%. After skin irritation test, it has no allergic reaction to sensitive skin people, and the scope of application has been greatly expanded. (3) Excellent storage stability: The polyol compound system significantly improves the stability of active ingredients. After 12 months of storage at room temperature, the activity retention rate is ≥92.7%, which solves the industry pain point of efficacy decay of beauty products; (4) Strong compatibility with cosmetics: The compound system takes into account both efficacy and skin feel. It is non-sticky when added to cosmetic products. It can be used in a ratio of 5% to 15% to match all types of repair products such as serums, face creams, and eye creams to achieve a full-chain skin care effect of "repair-moisturize-anti-aging". Detailed Implementation

[0017] Example 1 This embodiment uses the optimal parameters defined in the claims for preparation.

[0018] Raw material pretreatment and low-temperature fractional extraction: 10 kg of healthy, live white jade snails (70 g each) were selected and kept in a state of fasting and rest for 2.5 days (ambient temperature 21℃, relative humidity 68%), and rinsed once daily with sterile water at 25℃. 6 kg of soft tissue and 2 kg of foot gland secretions were aseptically separated; they were mixed at a ratio of soft tissue:foot gland secretions:sterile ice water = 3:1:10, and 80 L of sterile ice water was added to adjust the pH to 6.8; extraction was carried out at 4℃ with low-speed stirring for 2 hours, followed by further extraction at 15℃ with stirring for 1 hour. The extracts were combined to obtain 75 L of mixed extract.

[0019] Ultrasonic-assisted targeted hydrolysis with compound enzyme: 120g of compound enzyme preparation (papain:hyaluronidase = 2:1, total enzyme activity 1050U / mg) was added to the mixed extract, the pH was adjusted to 6.2, and the temperature was raised to 45℃; 20kHz ultrasound-assisted treatment was applied (power 300W, 5s working / 3s intermittent), and enzymatic hydrolysis was carried out at a constant temperature for 3.8h, with stirring for 5min every 30min (rate 50r / min); the enzyme was inactivated in a 75℃ water bath for 8min to obtain the enzymatic hydrolysate.

[0020] Membrane separation and purification: The enzymatic hydrolysate was centrifuged at 8000 rpm for 30 min at 4℃, and the supernatant was passed sequentially through a 5000 Da ultrafiltration membrane (pressure 0.3 MPa) and a 1000 Da nanofiltration membrane (pressure 0.55 MPa), with the feed temperature controlled at 6℃; the nanofiltration permeate was collected.

[0021] Steady-state formulation of active ingredients: Add a compounding agent consisting of glycerol, 1,3-propanediol and panthenol in a ratio of 5:3:2 to the nanofiltration permeate, with the addition amount being 12% of the weight of the permeate; stir at low speed at 25°C for 30 minutes to obtain a targeted skin care type white snail active complex.

[0022] Performance testing: The total purity of the active ingredients in this complex is 99.5%. After 12 months of storage at room temperature, the activity retention rate is 93.2%, and the skin fibroblast proliferation promotion rate is 53.8%. When added to face cream at a ratio of 10%, the product has a refreshing and non-sticky feel. After 28 days of use by volunteers with sensitive skin, the skin barrier repair rate reached 48.5%, and the dry lines were reduced by 32.7%.

[0023] Example 2 This embodiment uses the lower limit of the parameters defined in the claims for preparation to verify the robustness of the process.

[0024] Raw material pretreatment and low-temperature fractional extraction: 60g adult white jade snails were selected, fasted and rested for 3 days, and the ratio of soft tissue: foot gland secretions: sterile ice water was 3:1:10. Extraction was carried out at 4℃ for 2 hours and at 15℃ for 1 hour.

[0025] Ultrasonic-assisted directional hydrolysis with compound enzyme: enzyme dosage 1.2%, ultrasonic power 300W, hydrolysis time 3.5h, other parameters are the same as in Example 1.

[0026] Membrane separation purification: ultrafiltration pressure 0.25 MPa, nanofiltration pressure 0.5 MPa, feed temperature 8°C, other parameters are the same as in Example 1.

[0027] Steady-state compounding of active ingredients: 10% of compounding adjuvants were added, the stirring time was 30 min, and other parameters were the same as in Example 1.

[0028] Performance testing: The total purity of the active ingredients in the complex is 99.1%, the activity retention rate is 92.7% after 12 months of storage at room temperature, and the skin fibroblast proliferation promotion rate is 52.6%, which meets the application standards for beauty products.

[0029] Example 3 This embodiment uses an alternative compound additive system for preparation to verify the flexibility of the formulation.

[0030] Preliminary steps: Repeat steps 1 to 3 in Example 1 to obtain nanofiltration permeate.

[0031] Steady-state compounding of active ingredients: A mixture of glycerol, xylitol and vitamin B5 in a mass ratio of 4:4:2 was added to the nanofiltration permeate as a compounding aid, with the addition amount being 15% of the weight of the permeate; the mixture was stirred at low speed for 30 minutes at 25°C to obtain the complex.

[0032] Performance testing: The total purity of the active ingredients in this complex is 99.3%. After 12 months of storage at room temperature, the activity retention rate is 93.5%, and the stability is improved by more than 40% compared with before replacement. When it is added to the eye serum at a ratio of 8%, the depth of fine lines around the eyes is reduced by an average of 18.4% after 14 days of use by the subjects, and there is no eye irritation.

[0033] To more intuitively compare the process parameters and final product performance of the three embodiments, the following table is provided: Raw material weight (g) 70 60 70 Fasting and rest period (days) 2.5 3 2.5 Extraction temperature (°C) 4→15 4→15 4→15 Enzyme dosage (%) 1.5 1.2 1.5 Enzymatic hydrolysis time (h) 3.8 3.5 3.8 Compound adjuvant system Glycerin:Propylene Glycol:Panthenol (5:3:2) Glycerin:Propylene Glycol:Panthenol (5:3:2) Glycerin:Xylitol:VB5 (4:4:2) Compound additives addition amount (%) 12 10 15 Total purity of active ingredients (%) 99.5 99.1 99.3 Room temperature retention rate (12 months, %) 93.2 92.7 93.5 Cell proliferation promotion rate (%) 53.8 52.6 52.9 Main application effects Barrier repair rate: 48.5%; Fine lines reduced rate: 32.7%. Compliant with application standards Fine lines around the eyes reduced by 18.4% The present invention and its embodiments have been described above. This description is not restrictive, and what is shown is only one embodiment of the present invention. The actual structure is not limited to this. In conclusion, if those skilled in the art are inspired by this description and design similar structures and embodiments without departing from the spirit of the present invention, they should all fall within the protection scope of the present invention.

Claims

1. A process for the preparation of a targeted care-type helix aspersa active complex, characterized in that, Includes the following steps: Step S1, Raw Material Pretreatment and Low-Temperature Fractional Extraction: Select healthy, live white jade snails weighing 60-80g and place them in an environment with a temperature of 20-22℃ and a relative humidity of 65%-70% for 2-3 days without food. Aseptically separate the soft tissue and foot gland secretions, and mix them at a mass ratio of soft tissue:foot gland secretions:sterile ice water = 3:1:

10. Adjust the pH of the system to 6.5-7.

0. First, extract the snails by stirring at low speed at 4℃ for 2 hours to extract glycosaminoglycans and growth factors. Then, raise the temperature to 15℃ and continue stirring for 1 hour to extract snail enzymes. Combine the two extracts to obtain a mixed extract. Step S2, Ultrasonic-Assisted Targeted Hydrolysis with Compound Enzyme: Add a compound enzyme preparation to the mixed extract. The compound enzyme preparation consists of papain and hyaluronidase in a mass ratio of 2:1, with a total enzyme activity ≥1000 U / mg. The amount of enzyme added is 1.2%–1.8% of the total weight of the raw materials. Adjust the pH of the system to 6.0–6.5, raise the temperature to 45°C, and apply 20kHz ultrasound-assisted treatment at a power of 300W, working for 5 seconds and then intermittently for 3 seconds, for isothermal hydrolysis for 3.5–4 hours. After hydrolysis, inactivate the enzyme in a 75°C water bath for 8 minutes to obtain the hydrolysate. Step S3, Membrane separation and purification: The enzymatic hydrolysate is centrifuged at 4℃ and 8000rpm for 30min. The supernatant is then sequentially purified by passing it through a 5000Da ultrafiltration membrane and a 1000Da nanofiltration membrane. The ultrafiltration pressure is 0.25-0.35MPa, the nanofiltration pressure is 0.5-0.6MPa, and the feed temperature is controlled at 4-8℃. The 1000Da nanofiltration permeate is collected to obtain the core active ingredient solution of white jade snail. Step S4, Steady-state compounding of active ingredients: Add compounding adjuvants to the core active ingredient solution. The compounding adjuvants are composed of glycerin, 1,3-propanediol and panthenol in a mass ratio of 5:3:

2. The amount of compounding adjuvants added is 10% to 15% of the weight of the active ingredient solution. Stir and mix at low speed for 30 minutes at 25°C to obtain a targeted skin care type white snail active complex. Step S5, Aseptic filling and storage: After aseptic filtration of the complex through a 0.22μm microporous membrane, it is vacuum-filled in the dark and stored at 4–8℃.

2. The process for preparing a targeted skin care type of Eubania vermiculata active complex according to claim 1, characterized in that: In step S1, the white jade snails should be selected from adult snails with intact shells and clear crawling tracks. During the fasting and quiet recuperation period, they should be rinsed once a day with sterile water at 25°C to remove feces and impurities.

3. The process for preparing a targeted skin care type of Eubranchus sp. active complex according to claim 1, characterized by: The power density of the ultrasonic co-processing in step S2 is 0.55 W / cm², and the enzyme hydrolysis is stirred once every 30 min at a stirring rate of 50 r / min for 5 min.

4. The process for preparing a targeted skin care type of Eubranchus sp. active complex according to claim 1, characterized by: The 5000Da ultrafiltration membrane mentioned in step S3 is used to retain large molecular proteins, and the permeate of the 1000Da nanofiltration membrane contains snail enzyme content ≥18mg / g, glycosaminoglycan content ≥25mg / g, and growth factor content ≥12mg / mL.

5. The process for preparing a targeted skin care type of Eubranchus sp. active complex according to claim 1, characterized by: The compounding agent mentioned in step S4 can be replaced by a mixed system of glycerol, xylitol and vitamin B5 in a mass ratio of 4:4:

2. After compounding, the stability of the active ingredients at room temperature is improved by more than 40%.

6. The process for preparing a targeted skin care type of Eubranchus sp. active complex according to claim 1, characterized by: The process achieves a comprehensive extraction rate of ≥95.3% for three core components: snail enzyme, glycosaminoglycan, and growth factors.

7. A process for the preparation of a targeted skin care type of E. albolabris active complex according to any one of claims 1 to 6, characterized by: The total purity of the snail enzyme, glycosaminoglycan, and growth factors in the complex is ≥99.1%. After 12 months of storage at room temperature, the retention rate of active ingredients is ≥92.7%, the proliferation promotion rate of skin fibroblasts is ≥52.6%, and there are no sensitizing impurities remaining.

8. The process for preparing a targeted skin care type of Eubranchus sp. active complex according to claim 7, characterized by: The complex exhibits optimal stability in the pH range of 5.5–6.5 and has an electrical conductivity of 200–400 μS / cm.

9. The preparation process of a targeted skincare-enhancing white jade snail active complex according to claim 7 or 8, characterized in that: The complex is added to serums, face creams, eye creams, and body lotions at a mass percentage of 5% to 15% for skin barrier repair, fine line reduction, and sensitive skin soothing.

10. The process for preparing a targeted skin care type of Eubranchus sp. active complex according to claim 9, characterized by: When applied to face cream, the addition amount is 10%. After 28 days of use, the skin barrier repair rate is ≥48.5%, and the dry lines are reduced by ≥32.7%.