Molecular weight gradient directional freeze-drying essence composition as well as preparation method and application thereof

Abstract

The invention relates to the field of compositions, in particular to a molecular weight gradient directional freeze-drying essence composition as well as a preparation method and application thereof. The molecular weight gradient directional freeze-dried essence composition provided by the invention is prepared from the following raw materials: small molecular weight polypeptide, medium molecular weight polysaccharide and high molecular weight polysaccharide; wherein the molecular weight of the polypeptide with the small molecular weight is 200-5000Da, the molecular weight of the polysaccharide with the medium molecular weight is 10000-1000000Da, and the molecular weight of the polysaccharide with the high molecular weight is 1050000-2300000Da; the preparation process relates to a pre-cooling stage, a phase separation stage and a deep freezing stage. The molecular weight gradient directional freeze-dried essence composition has excellent continuous permeability and high storage stability, and provides milder and more comfortable product use experience for consumers.

Description

Technical Field

[0001] The present invention relates to the field of compositions, and specifically, to a molecular weight gradient-directed freeze-dried essence composition, its preparation method and application. Background Art

[0002] As a new type of skin care technology, freeze-dried masks have been widely used in the cosmetics field in recent years. Compared with traditional mask liquids, freeze-dried masks efficiently preserve active ingredients through freeze-drying technology, avoiding the use of preservatives, and thus are safer for the skin. In addition, freeze-dried masks have good stability at room temperature, can significantly extend the shelf life of products, and maintain the integrity of active ingredients. These characteristics have enabled them to rapidly rise in the skin care product market and are especially favored by consumers who pursue efficient and safe skin care.

[0003] Although the freeze-dried mask technology has significant advantages, there are generally some technical bottlenecks in existing products. Ordinary freeze-dried masks mostly adopt a homogenized formulation design. For example, Chinese Patent Application CN119326680A discloses a freeze-dried mask, the preparation raw materials of which include deionized water, algae extract, mannitol, glycerol, hydrolyzed collagen, amino acid mixture, hydroxypropyltetrahydropyran triol, ectoine, and hyaluronic acid; the preparation method includes uniformly mixing all raw materials together to obtain a freeze-dried mask essence solution, and then freeze-drying. However, this freeze-drying process is difficult to achieve the technical effects of long-acting penetration, non-irritation, and continuous moisturization of the freeze-dried mask.

[0004] Therefore, there is an urgent need for a freeze-dried essence composition with long-acting penetration, continuous moisturization, non-irritation, and excellent beauty effects to fill the relevant technical gaps. Summary of the Invention

[0005] To solve the above technical problems, the first aspect of the present invention provides a molecular weight gradient-directed freeze-dried essence composition, the preparation raw materials of which include small molecular weight polypeptides, medium molecular weight polysaccharides, and high molecular weight polysaccharides; the molecular weight of the small molecular weight polypeptides is 200 - 5000 Da, the molecular weight of the medium molecular weight polysaccharides is 10000 - 1000000 Da, and the molecular weight of the high molecular weight polysaccharides is 1050000 - 2300000 Da.

[0006] As an implementable case, the mass ratio of the small molecular weight polypeptides, medium molecular weight polysaccharides, and high molecular weight polysaccharides is (0.01 - 0.05) : (0.05 - 0.2) : (0.05 - 0.2).

[0007] Furthermore, the small molecular weight polypeptides include at least two of carnosine, acetyl hexapeptide-8, arginine / lysine polypeptide, or palmitoyl pentapeptide-4.

[0008] Furthermore, the small molecular weight polypeptides include carnosine, acetyl hexapeptide-8, arginine / lysine polypeptide, and palmitoyl pentapeptide-4; the mass ratio of carnosine, acetyl hexapeptide-8, arginine / lysine polypeptide, and palmitoyl pentapeptide-4 is (0.001 - 0.01):(0.001 - 0.005):(0.0001 - 0.001):(0.0001 - 0.001).

[0009] Furthermore, the mass ratio of carnosine, acetyl hexapeptide-8, arginine / lysine polypeptide, and palmitoyl pentapeptide-4 is 0.01:0.003:0.0005:0.0005.

[0010] As an implementable case, the medium molecular weight polysaccharides include one or more of: β-glucan, hyaluronic acid, sodium hyaluronate, tremella polysaccharide, seabuckthorn polysaccharide, yeast polysaccharide, ganoderma lucidum polysaccharide, or seaweed polysaccharide.

[0011] Further, the medium molecular weight polysaccharide is tremella polysaccharide, and its molecular weight is 1000000 Da.

[0012] As an implementable case, the high molecular weight polysaccharides include one or more of: β-glucan, hyaluronic acid, sodium hyaluronate, tremella polysaccharide, seabuckthorn polysaccharide, yeast polysaccharide, ganoderma lucidum polysaccharide, or seaweed polysaccharide.

[0013] Further, the high molecular weight polysaccharide is sodium hyaluronate, and its molecular weight is 1600000 Da.

[0014] In this application, small-molecular-weight polypeptides include carnosine, acetyl hexapeptide-8, arginine / lysine polypeptide, and palmitoyl pentapeptide-4, which can quickly penetrate deep into the skin, transmit biological signals, promote collagen synthesis, inhibit melanin production, etc., and exert anti-wrinkle and whitening effects; medium-molecular-weight polysaccharide tremella polysaccharide can form a network structure in the middle layer of the skin, lock in moisture, provide long-lasting moisturization, and at the same time regulate the skin microenvironment and enhance the skin's self-repair ability. High-molecular-weight sodium hyaluronate can form a protective film on the skin surface, lock in moisture, prevent moisture loss, and at the same time cooperate with other ingredients to enhance the overall moisturizing effect. Through the molecular weight gradient orientation system, small-molecular-weight polypeptides can quickly penetrate deep into the skin, medium-molecular-weight polysaccharides play a role in the middle layer, and high-molecular-weight polysaccharide sodium hyaluronate forms a protective film on the surface layer. The three cooperate to enable the active substances to penetrate in sequence, greatly improving the penetration efficiency, and the continuous penetration time reaches 6 hours; in addition, small-molecular-weight polypeptides as active substances, medium-molecular-weight polysaccharides and sodium hyaluronate as moisturizers, under their combined action, the moisturizing and penetration-promoting effects promote each other. The molecular weight gradient system can control the penetration speed and depth of the active substances, reduce the irritation reaction caused by premature penetration of small-molecular active substances, and avoid excessive accumulation of macromolecular moisturizers in the stratum corneum. At the same time, the compound system can also enable the active substances to better exert anti-wrinkle, moisturizing, soothing and other effects.

[0015] The second aspect of the present invention provides a preparation method of a molecular weight gradient orientation freeze-dried essence composition, including:

[0016] S1. Pre-cooling stage: Mix small-molecular-weight polypeptides, medium-molecular-weight polysaccharides, and high-molecular-weight polysaccharides evenly, then place them in a freeze dryer, cool down to -8 to -3°C, and then keep warm for 25 to 35 minutes;

[0017] S2. Phase separation stage: Continue to cool down to -30 to -20°C and keep warm for 10 to 14 minutes;

[0018] S3. Deep freezing stage: Continue to cool down to -50 to -42°C and keep warm for 5 to 7 minutes to obtain the molecular weight gradient orientation freeze-dried essence composition.

[0019] As an implementable case, in the step S1, the cooling rate is 1 - 3°C / min.

[0020] As an implementable case, in the step S2, the cooling rate is 3 - 5°C / min.

[0021] As an implementable case, in the step S3, the cooling rate is 8 - 10°C / min.

[0022] Furthermore, in the step S1, the cooling rate is 2°C / min.

[0023] Further, in the step S2, the cooling rate is 5 °C / min.

[0024] Further, in the step S3, the cooling rate is 10 °C / min.

[0025] In the process of preparing the freeze-dried essence composition of the present application, a stepwise cooling procedure is adopted. In the pre-cooling stage, the freeze-dried essence composition is initially cooled to provide a mild starting environment for the active ingredients, prevent structural damage caused by rapid freezing, and at the same time help to initially fix the surface active ingredients to prepare for subsequent phase separation; in the phase separation stage, it is heat-insulated at -30 °C to -20 °C for 10 to 14 minutes to promote the phase separation of components with different molecular weights, form a molecular weight gradient distribution, small molecular weight polypeptides migrate to the deep layer, while medium molecular weight polysaccharide tremella polysaccharide and high molecular weight polysaccharide sodium hyaluronate aggregate in the middle layer and the surface layer respectively, realizing phase separation and forming a gradient distribution; finally, in the deep-freezing stage, the composition is rapidly deep-frozen at a cooling rate of 10 °C / min to fix the molecular weight gradient distribution, maintain the stability and biological activity of the active ingredients, and form a stable three-dimensional structure. The three-stage freezing procedure cooperates with each other. By precisely controlling the temperature and rate at different stages, not only a molecular weight gradient directional system is constructed to enable the active ingredients to penetrate in the expected order, but also the penetration efficiency and moisturizing effect are improved, and at the same time, the product stability and safety are significantly enhanced. After testing, the continuous penetration time of the freeze-dried essence composition can reach 6 hours, and the retention rate of the active substances is still above 95% after 6 months of storage.

[0026] The third aspect of the present invention provides an application of a molecular weight gradient directional freeze-dried essence composition, including the application in the preparation of a freeze-dried mask. Specifically, the freeze-dried essence composition, emulsifier, thickener and other raw materials are mixed evenly until the total mass fraction of the raw materials is 100 parts, and then attached to the dry film cloth to prepare the freeze-dried mask.

[0027] Beneficial effects

[0028] (1) By using three raw materials with different molecular weights, namely small molecular weight (200 - 5000 Da) polypeptides, medium molecular weight (10000 - 1000000 Da) polysaccharides and high molecular weight (1050000 - 2300000 Da) polysaccharides, the present application constructs an innovative molecular weight gradient directional system. This system can precisely control the penetration order of active substances, achieve the rapid penetration of small molecular weight polypeptides to the deep layer of the skin, the medium molecular weight polysaccharides play a role in the middle layer, and the high molecular weight polysaccharides form a protective film on the surface layer of the skin. The orderly penetration mechanism significantly improves the overall transdermal efficiency and ensures the effective delivery and utilization of active ingredients.

[0029] (2) The preparation process of the freeze-dried essence composition of the present invention includes three key stages: precooling, phase separation, and deep freezing. The gradient freeze-drying process can effectively protect the active ingredients from damage under different temperature and time conditions, ensuring the stability of the product under normal temperature storage conditions and the persistence of its efficacy. By precisely controlling the process parameters of each stage, not only the physicochemical properties of the product are optimized, but also its performance during long-term storage is enhanced.

[0030] (3) In the present invention, the design of the molecular weight gradient system effectively avoids the skin irritation problems that may be caused by the premature penetration of small molecular active substances, and at the same time prevents the excessive accumulation of macromolecular moisturizers in the stratum corneum, thus significantly improving the safety of the product and providing consumers with a more gentle and comfortable use experience.

[0031] (4) The freeze-dried essence composition of the present invention has excellent comprehensive efficacy, and can effectively anti-wrinkle, deeply moisturize, repair skin damage, and soothe skin discomfort. This is mainly due to the synergistic effect between the active ingredients, providing consumers with a comprehensive skin care solution.

[0032] (5) The freeze-dried essence composition of the present invention breaks through the technical bottleneck of the mutual restriction between the moisturizing and penetration-promoting effects in the traditional formula. It can not only achieve long-term moisturization, but also promote the penetration of active ingredients into the dermis layer, achieving the synergistic effect of both. This not only enables the product to moisturize while more efficiently delivering active ingredients, thus performing more excellently in improving the skin health state. Specific Embodiments

[0033] Example 1

[0034] In the first aspect of this example, a molecular weight gradient directional freeze-dried essence composition is provided. The preparation raw materials are as follows by mass fraction: 0.014 parts of small molecular weight polypeptides, 0.05 parts of medium molecular weight polysaccharides, and 0.05 parts of high molecular weight polysaccharides.

[0035] The specific small molecular weight polypeptides are: carnosine, acetyl hexapeptide-8, arginine / lysine polypeptide, and palmitoyl pentapeptide-4; the mass ratio of carnosine, acetyl hexapeptide-8, arginine / lysine polypeptide, and palmitoyl pentapeptide-4 is 0.01:0.003:0.0005:0.0005.

[0036] The medium molecular weight polysaccharide is tremella polysaccharide.

[0037] The high molecular weight polysaccharide is sodium hyaluronate with a molecular weight of 1,600,000 Da.

[0038] In the second aspect of this example, a preparation method of a molecular weight gradient directional freeze-dried essence composition is provided, specifically as follows:

[0039] S1. Pre-cooling stage: Mix small molecular weight polypeptides, medium molecular weight polysaccharides, and high molecular weight polysaccharides evenly, then place them in a freeze dryer, cool down to -5°C at a rate of 2°C / min, and then hold for 30 min;

[0040] S2. Phase separation stage: Continue to cool down to -25°C at a rate of 5°C / min, and then hold for 12 min;

[0041] S3. Deep freezing stage: Continue to cool down to -42°C at a rate of 10°C / min, hold for 6 min, and thus obtain the molecular weight gradient directional freeze-dried essence composition.

[0042] In the third aspect of this example, an application of the molecular weight gradient directional freeze-dried essence composition is provided, specifically applied to the preparation of freeze-dried masks.

[0043] Comparative Example 1

[0044] In the first aspect of this example, a molecular weight gradient directional freeze-dried essence composition is provided. The preparation raw materials are as follows by mass fraction: 0.014 parts of small molecular weight polypeptides, 0.05 parts of medium molecular weight polysaccharides, and 0.05 parts of high molecular weight polysaccharides.

[0045] The specific small molecular weight polypeptides are: carnosine, acetyl hexapeptide-8, arginine / lysine polypeptide, and palmitoyl pentapeptide-4; the mass ratio of carnosine, acetyl hexapeptide-8, arginine / lysine polypeptide, and palmitoyl pentapeptide-4 is 0.01:0.003:0.0005:0.0005.

[0046] The medium molecular weight polysaccharide is tremella polysaccharide.

[0047] The high molecular weight polysaccharide is sodium hyaluronate with a molecular weight of 1,600,000 Da.

[0048] In the second aspect of this example, a preparation method of the molecular weight gradient directional freeze-dried essence composition is provided, specifically as follows:

[0049] Mix small molecular weight polypeptides, medium molecular weight polysaccharides, and high molecular weight polysaccharides evenly, then place them in a freeze dryer, cool down to -40°C at a rate of 15°C / min, and then hold for 30 min;

[0050] In the third aspect of this example, an application of the molecular weight gradient directional freeze-dried essence composition is provided, specifically applied to the preparation of freeze-dried masks.

[0051] Comparative Example 2

[0052] The specific implementation manner of this example is the same as that of Example 1, the difference is that the molecular weight of the sodium hyaluronate is 300,000 Da, belonging to medium molecular weight polysaccharides.

[0053] Comparative Example 3

[0054] In the first aspect of this example, a molecular weight gradient directional freeze-dried essence composition is provided, and the raw materials for its preparation are specifically 0.014 parts of low molecular weight polypeptides and 0.1 parts of medium molecular weight polysaccharides in terms of weight fractions.

[0055] The small molecular weight polypeptides are specifically: carnosine, acetyl hexapeptide-8, arginine / lysine polypeptide and palmitoyl pentapeptide-4; the mass ratio of carnosine, acetyl hexapeptide-8, arginine / lysine polypeptide and palmitoyl pentapeptide-4 is 0.01:0.003:0.0005:0.0005.

[0056] The medium molecular weight polysaccharide is Tremella polysaccharide.

[0057] The second aspect of this example provides a method for preparing a molecular weight gradient directional freeze-dried essence composition, specifically:

[0058] S1, pre-cooling stage: small molecular weight polypeptides and medium molecular weight polysaccharides are mixed evenly, then placed in a freeze dryer, cooled to -5°C at a rate of 2°C / min, and then kept warm for 30 minutes;

[0059] S2, phase separation stage: continue cooling to -25°C at a rate of 5°C / min, and then keep warm for 12 minutes;

[0060] S3, deep freezing stage: continue to cool to -42°C, the cooling rate is 10°C / min, and keep warm for 6 minutes to obtain a molecular weight gradient directional freeze-dried essence composition.

[0061] The third aspect of this example provides an application of a molecular weight gradient directional freeze-dried essence composition, which is specifically applied to the preparation of a freeze-dried facial mask.

[0062] The molecular weight and source information of the raw materials in Example 1 and Comparative Examples 1-3 are shown in Table 1.

[0063] Table 1

[0064]

[0065] Performance Evaluation

[0066] 1. Penetration performance test

[0067] Test samples: molecular weight gradient directional freeze-dried essence compositions prepared in Example 1 and Comparative Examples 1-3.

[0068] Test object: human epidermis.

[0069] Test method: Franz diffusion cell method;

[0070] Apparatus: TK-20 Automatic Sampling Franz Diffusion Cell System;

[0071] Diffusion area: 2.0 cm 2 ;

[0072] Volume of the receiving cell: 12.5 mL;

[0073] Temperature: 32 ± 0.5 °C;

[0074] Stirring speed: 600 rpm;

[0075] Sampling time points: 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h;

[0076] Detection method: Quantitative analysis by UPLC-MS / MS;

[0077] Reference standards:

[0078] 1. "Regulations on the Administration of Cosmetics New Raw Material Registration and Filing Information";

[0079] 2. Appendix A of "Technical Specifications for Cosmetics Safety" (2015 Edition);

[0080] In this experiment, through the Franz diffusion cell method, the penetration characteristics of the molecular weight gradient-directed freeze-dried combined essence prepared in Example 1 and Comparative Examples 1-3 on the human epidermis were studied comparatively, including the total percutaneous amount in 8 h, the dermal layer arrival rate, the lag time of penetration, and the sustained penetration time, to evaluate the transdermal absorption effect of different formula compositions. The experimental results are shown in Table 2.

[0081] Table 2

[0082] Test Items Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 Total Transdermal Amount in 8h <![CDATA[682±45μg / cm 2 > <![CDATA[205±28μg / cm 2 > <![CDATA[385±32μg / cm 2 > <![CDATA[82±15μg / cm 2 > Reaching Rate of Dermis 58％ 12％ 24％ 6％ Penetration Lag Time 1.8 min 4.5 min 3.2 min 6.8 min Continuous Penetration Time 6h 2.1h 3.6h 1.1h

[0083] The experimental results in Table 2 show that the molecular weight gradient-directed freeze-dried essence composition prepared in Example 1 has excellent sustained penetration effect, the sustained penetration time can reach 6 h, and the total percutaneous amount in 8 h reaches 500 μg / cm 2 Above, the penetration effect is excellent.

[0084] 2. Moisturizing-Penetration Promoting Synergistic Effect Test

[0085] Test samples: The molecular weight gradient-directed freeze-dried essence compositions prepared in Example 1 and Comparative Examples 1-3.

[0086] Test instrument: Corneometer CM 825 Capacitance Method Moisture Tester;

[0087] Test environment: Temperature 22 ± 2 °C, relative humidity 50 ± 5%;

[0088] Test site: Specified areas of the face, 3 points are measured in each area;

[0089] Equilibration time: 20 min;

[0090] Recording time points: 0.5 h, 2 h, 6 h;

[0091] Test method for active substance concentration:

[0092] Test instrument: Agilent 1290 UPLC tandem AB SCIEX 4500 mass spectrometer;

[0093] Chromatographic column: Waters ACQUITY UPLC BEH C18 (2.1×100 mm, 1.7 μm);

[0094] Mobile phase: Phase A is 0.1 wt% formic acid aqueous solution, and phase B is acetonitrile;

[0095] Flow rate: 0.3 mL / min;

[0096] Column temperature: 40 °C;

[0097] Injection volume: 2 μL.

[0098] Reference standards:

[0099] 1. QB / T 2872-2017 "Test Method for Moisture in Cosmetics";

[0100] 2. GB / T 35892-2018 "High Performance Liquid Chromatography Method for the Determination of Active Ingredient Content in Cosmetics".

[0101] In this experiment, instruments such as the Corneometer CM 825 capacitance method moisture tester and the Agilent 1290 UPLC tandem ABSCIEX 4500 mass spectrometer were used to conduct a test on the moisturizing and permeability-enhancing synergistic effects of the molecular weight gradient-directed freeze-dried essence compositions prepared in Example 1 and Comparative Examples 1-3. The test was carried out in an environment with a temperature of 22±2 °C and a relative humidity of 50±5%. Specified areas of the face and other parts were selected, 3 points were measured in each area. After a 20-min equilibration time, detections were carried out at the recording time points of 0.5 h, 2 h, and 6 h to analyze the changes in moisture content and the concentration of active substances, and then evaluate the moisturizing and permeability-enhancing effects of each composition. The experimental results are shown in Table 3.

[0102] Table 3

[0103]

[0104] The experimental results in Table 3 show that the molecular weight gradient - oriented freeze - dried essence composition prepared in Example 1 has excellent moisturizing - promoting penetration synergistic effect. The water content of the stratum corneum can reach 65% after 6 hours, and the concentration of active substances in the dermis layer can reach 9.8 μg / cm 3 .

[0105] 3. Safety Test

[0106] Test samples: The molecular weight gradient - oriented freeze - dried essence compositions prepared in Example 1, Comparative Examples 1 - 3.

[0107] Test method: Human patch test;

[0108] Subjects: 50 healthy volunteers (including 20 cases of sensitive skin);

[0109] Application method: Finn chamber closed application;

[0110] Application time: 24 h;

[0111] Observation time points: Immediately after application (0 h), 24 h, 48 h;

[0112] Scoring criteria: The standard of the International Contact Dermatitis Research Group (ICDRG);

[0113] Reference standard: GB / T 29665 - 2013 "Cosmetics Skin Irritation Test".

[0114] In this experiment, the human patch test was used to conduct safety tests on the molecular weight gradient - oriented freeze - dried essence compositions prepared in Example 1 and Comparative Examples 1 - 3. Fifty healthy volunteers (including 20 cases of sensitive skin) were selected as subjects. Using the Finn chamber closed application method, after application for 24 h, according to the standard of the International Contact Dermatitis Research Group (ICDRG), observations and scoring were carried out immediately after application (0 h), 24 h, and 48 h. Evaluation was based on the standard of GB / T 29665 - 2013 "Cosmetics Skin Irritation Test" to investigate the impact of the test substances on skin adverse reactions. The experimental results are shown in Table 4 for details.

[0115] Table 4

[0116] Stimulation Type / Incidence Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 Instantaneous Prick 4.7％ 18.3％ 12.5％ 25.6％ Persistent Erythema 0％ 6.2％ 3.8％ 8.9％ Irritant Dermatitis 0％ 2.8％ 1.2％ 4.5％

[0117] The experimental results in Table 4 show that the molecular weight gradient - oriented freeze - dried essence composition prepared in Example 1 has excellent use safety. The probability of instantaneous stinging is less than 5%, and the occurrence probabilities of persistent erythema and irritant dermatitis are 0%. The freeze - dried essence composition is also effective for sensitive skin; while the safety performance of the freeze - dried essence composition in Comparative Example 3 without high - molecular - weight polysaccharides is relatively the worst.

[0118] 4. Efficacy Test

[0119] Test samples: Molecular weight gradient-oriented freeze-dried essence compositions prepared in Example 1 and Comparative Examples 1-3.

[0120] Wrinkle depth test method:

[0121] Test site: Human epidermis;

[0122] Test instrument: PRIMOS skin three-dimensional image analysis system;

[0123] Test parameters: Ra (average roughness), Rz (maximum profile height);

[0124] Scanning area: 30×40 mm;

[0125] Resolution: 0.1 μm vertically and 0.06 mm horizontally;

[0126] Transepidermal water loss test method:

[0127] Test site: Human epidermis;

[0128] Instrument: Tewameter TM300;

[0129] Environmental conditions: Temperature 22±2°C, relative humidity 50±5%;

[0130] Test time: 60 s per point;

[0131] Data collection: Once per second;

[0132] Skin elasticity test method:

[0133] Test site: Human epidermis;

[0134] Instrument: Cutometer MPA580;

[0135] Test mode: Suction pressure 450 mbar;

[0136] Suction time: 2 s, release time: 2 s;

[0137] Number of repetitions: 3 times per test point;

[0138] Reference standards:

[0139] 1. GB / T 38848-2020 "Test Method for Evaluating the Efficacy of Cosmetics in Improving Wrinkles";

[0140] 2. QB / T 2873-2017 "Test Method for Transepidermal Water Loss (TEWL) of Cosmetics";

[0141] 3. "Test Method for Skin Elasticity Evaluation of Cosmetics Efficacy" (GB / T 38847-2020).

[0142] In this experiment, the molecular weight gradient directional freeze-dried essence compositions of Example 1 and Comparative Examples 1-3 were tested for efficacy using a variety of instruments. For wrinkle depth testing, a PRIMOS skin three-dimensional image analysis system was used. The test site was the human epidermis, and the parameters were Ra (average roughness) and Rz (maximum profile height). The scanning area was 30×40 mm, with a vertical resolution of 0.1 μm and a horizontal resolution of 0.06 mm. For the measurement of transepidermal water loss, a Tewameter TM300 was used. The test site was also the human epidermis, and the environmental conditions were a temperature of 22±2°C and a relative humidity of 50±5%. Each point was measured for 60 s, and data was collected once per second. For skin elasticity testing, a Cutometer MPA580 was used. The test mode was an aspiration pressure of 450 mbar, an aspiration time of 2 s, and a release time of 2 s. Each test point was repeated 3 times. The tests were all carried out according to relevant standards, including "Test Method for Wrinkles Improvement Evaluation of Cosmetics Efficacy" (GB / T 38848-2020), "Test Method for Transepidermal Water Loss (TEWL) of Cosmetics" (QB / T 2873-2017), and "Test Method for Skin Elasticity Evaluation of Cosmetics Efficacy" (GB / T 38847-2020); the experimental results are shown in Table 5.

[0143] Table 5

[0144] Efficacy Index / Improvement Rate Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 Wrinkle Depth 43％↓ 12％↓ 18％↓ 5％↓ Transepidermal Water Loss 62％↓ 25％↓ 38％↓ 15％↓ Skin Elasticity 58％↑ 18％↑ 27％↑ 8％↑ Redness Relief 79％↓ 32％↓ 45％↓ 12％↓

[0145] The experimental results in Table 5 show that the molecular weight gradient directional freeze-dried essence composition prepared in Example 1 has excellent performance in use. After use, it can effectively improve wrinkle depth, transepidermal water loss, skin elasticity, and redness relief, and the efficacy improvement performance is significantly better than that of Comparative Examples 1-3.

[0146] 5. Stability Test

[0147] Test samples: The molecular weight gradient directional freeze-dried essence compositions prepared in Example 1 and Comparative Examples 1-3.

[0148] Test method: Accelerated test;

[0149] Storage conditions: 40±2°C, relative humidity 75±5%;

[0150] Test time points: 0, 1, 3, 6 months;

[0151] Test items: Appearance, pH value, active ingredient content;

[0152] Determination of active ingredient content: UPLC-MS / MS method;

[0153] Reference standards:

[0154] 1. "Accelerated Test Methods for Cosmetics" (GB / T 31603-2015);

[0155] 2. "Determination Method for the Content of Efficacy Ingredients in Cosmetics - High Performance Liquid Chromatography" (YBB00102003-2015).

[0156] In this experiment, stability tests were conducted on the molecular weight gradient - oriented freeze - dried essence compositions of Example 1 and Comparative Examples 1 - 3 using the accelerated test method. The samples were placed under the conditions of 40 ± 2°C and relative humidity of 75 ± 5%. The test time points were 0, 1, 3, and 6 months. The detection items included appearance, pH value, and active ingredient content, and the active ingredient content was determined by UPLC - MS / MS method. The tests were carried out in accordance with "Accelerated Test Methods for Cosmetics" (GB / T 31603-2015) and "Determination Method for the Content of Efficacy Ingredients in Cosmetics - High Performance Liquid Chromatography" (YBB00102003-2015) to evaluate the stability of each composition under specific environmental conditions; the experimental results are shown in Table 6 for details.

[0157] Table 6

[0158] Storage Time / Active Ingredient Retention Rate Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 1 month 99.2％ 85.3％ 92.7％ 78.4％ 3 months 97.8％ 76.1％ 88.5％ 65.2％ 6 months 95.1％ 68.9％ 82.3％ 53.7％

[0159] The experimental results in Table 6 show that the molecular weight gradient - oriented freeze - dried essence composition prepared in Example 1 has excellent storage stability. After being stored for 6 months under the environment of 40 ± 2°C and relative humidity of 75 ± 5%, the retention rate of the active ingredient is still greater than 95%, while the storage stability of Comparative Examples 1 - 3 is relatively poor.

Claims

1. A molecular weight gradient-directed freeze-dried essence composition, characterized in that, The preparation raw materials include small molecular weight polypeptides, medium molecular weight polysaccharides, and high molecular weight polysaccharides; The molecular weight of the small molecular weight polypeptide is 200 - 5000 Da, the molecular weight of the medium molecular weight polysaccharide is 10000 - 1000000 Da, and the molecular weight of the high molecular weight polysaccharide is 1050000 - 2300000 Da.

2. The molecular weight gradient-directed freeze-dried essence composition according to claim 1, characterized in that, The mass ratio of the small molecular weight polypeptide, medium molecular weight polysaccharide, and high molecular weight polysaccharide is (0.01 - 0.05):(0.05 - 0.2):(0.05 - 0.2).

3. The molecular weight gradient-directed freeze-dried essence composition according to claim 1, wherein The small molecular weight polypeptide includes at least two of carnosine, acetyl hexapeptide - 8, arginine / lysine polypeptide, or palmitoyl pentapeptide - 4.

4. The molecular weight gradient-directed freeze-dried essence composition according to claim 1, characterized in that, The medium molecular weight polysaccharide includes one or more of β - glucan, hyaluronic acid, sodium hyaluronate, tremella polysaccharide, seabuckthorn polysaccharide, yeast polysaccharide, ganoderma lucidum polysaccharide, or seaweed polysaccharide.

5. The molecular weight gradient-directed freeze-dried essence composition according to claim 1, characterized in that, The high molecular weight polysaccharide includes one or more of β - glucan, hyaluronic acid, sodium hyaluronate, tremella polysaccharide, seabuckthorn polysaccharide, yeast polysaccharide, ganoderma lucidum polysaccharide, or seaweed polysaccharide.

6. A preparation method of the molecular weight gradient-directed freeze-dried essence composition according to any one of claims 1-5, characterized in that, It includes the following steps: S1. Pre - cooling stage: Mix the small molecular weight polypeptide, medium molecular weight polysaccharide, and high molecular weight polysaccharide evenly, then place them in a freeze - dryer, cool down to - 8~ - 3℃, and then keep warm for 25~35 min; S2. Phase separation stage: Continue to cool down to - 30~ - 20℃ and keep warm for 10~14 min; S3. Deep - freezing stage: Continue to cool down to - 50~ - 42℃ and keep warm for 5~7 min to obtain the molecular weight gradient - oriented freeze - dried essence composition.

7. The preparation method of the molecular weight gradient oriented freeze-dried essence composition according to claim 6, characterized in that, In the S1 step, the cooling rate is 1 - 3℃ / min.

8. The preparation method of the molecular weight gradient-oriented freeze-dried essence composition according to claim 6, characterized in that, In the S2 step, the cooling rate is 3 - 5℃ / min.

9. The preparation method of the molecular weight gradient-directed freeze-dried essence composition according to claim 6, wherein In the S3 step, the cooling rate is 8 - 10℃ / min.

10. Use of the molecular weight gradient-directed freeze-dried essence composition according to any one of claims 1-5, characterized in that, It is applied to the preparation of freeze - dried facial masks.

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