Recombinant albumin anti-aging composition as well as preparation method and application thereof

Abstract

The invention relates to the field of cosmetics, in particular to a recombinant albumin anti-aging composition as well as a preparation method and application thereof. The composition provided by the invention comprises the following components in parts by mass: 0.1-15.0 parts of recombinant human albumin; 0.1 to 10.0 parts of a grape wine vinasse polysaccharide extract; the content of the red raspberry extract is 0.1 to 5.0 parts. According to the composition disclosed by the invention, the recombinant human albumin, the wine vinasse polysaccharide extract and the red raspberry extract have a synergistic effect, so that the composition has the effects of inhibiting the activity of tyrosinase, promoting regeneration of collagen and elastin, tightening and resisting wrinkles and effectively resisting aging, and the composition disclosed by the invention is simple in component, good in biocompatibility and free of toxic and side effects. The sensitization of the wine vinasse extract and the red raspberry extract is improved, and the method can be widely applied to preparation of anti-aging products.

Description

Technical Field

[0001] The present invention relates to the field of cosmetics, and in particular to a recombinant albumin anti-aging composition and a preparation method and application thereof. Background Art

[0002] As we age, factors such as lifestyle and sun exposure lead to a decrease in skin collagen, which causes facial skin aging, dullness, loss of elasticity, spots, wrinkles, etc. Related cosmetics are emerging in an endless stream, and facial masks have become an indispensable item in people's daily lives because of their convenience and effectiveness. However, the active ingredients of most facial masks on the market are made through chemical synthesis, which may produce certain effects in the short term, but long-term use will endanger human health.

[0003] Grape lees: Grape lees refer to the grape residue filtered out during the winemaking process. Grape lees are mainly composed of dead yeast cells, grape seeds, grape skins, grape stems and insoluble tartaric acid, which are usually precipitated during the winemaking and aging process. Grape lees are rich in protein amino acids, active peptides, functional oligosaccharides, antioxidant phenols and flavonoids, as well as other functional ingredients and various trace elements.

[0004] Red raspberry extract: Red raspberry (Rubus idaeus L.) is a perennial small shrub fruit tree of the genus Rubus (Rubus spp) of the family Rosaceae, also known as raspberry, mountain raspberry, and raspberry. Red raspberry originated in Europe, and Russia is the earliest country to cultivate red raspberry. It is currently widely planted in Europe, Asia and America. It contains rich nutrients and bioactive ingredients, such as sugars, vitamins, minerals, anthocyanins, ellagic acid, raspberry ketones, etc. It has multiple functions such as anti-oxidation, anti-bacterial, anti-aging, anti-inflammatory, anti-cancer, immunity enhancement, heart protection, and weight loss. It is widely used in food, medicine, and health products.

[0005] However, wine lees extract and red raspberry extract are more often used in health products or food fields due to their allergenicity, and there is relatively little research on them in the cosmetics field. Summary of the invention

[0006] In view of the deficiencies in the prior art, the present invention applies wine lees and red raspberries to the cosmetics field, and provides an anti-aging composition and a preparation method and application thereof. The specific contents of the present invention are as follows: In a first aspect, the present invention provides an anti-aging composition, comprising: 0.1 to 15.0 parts of recombinant human albumin; 0.1 to 10.0 parts of wine lees polysaccharide extract; and 0.1 to 5.0 parts of red raspberry extract.

[0007] Furthermore, the composition also includes additives acceptable in the field of cosmetics. Further, the additives include at least one of preservatives, emulsifiers, humectants, thickeners, pH regulators, surfactants, stabilizers, antioxidants, and moisturizers.

[0008] Furthermore, the additive includes at least one of phenoxyethanol, parabens, vitamin E (tocopherol), butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), polysorbate (Tween series), glyceryl stearate, carbomer, xanthan gum, glycerin, citric acid, sodium hydroxide, sodium lauryl sulfate (SLS), cocamidopropyl betaine, ethylenediaminetetraacetic acid (EDTA), and ethylhexyl palmitate.

[0009] In a specific embodiment of the present invention, the additives include glycerol, carbomer, ethylhexyl palmitate, and polysorbate 80.

[0010] It should be noted that the present invention does not strictly limit the composition and weight fraction of additives acceptable in the field of cosmetics. All one or more additives with the functions of preservatives, emulsifiers, humectants, thickeners, pH adjusters, surfactants, stabilizers, antioxidants, and moisturizers that are harmless to the human body and are selected according to the dosage form of the product, or the weight fraction of the additives is routinely adjusted according to actual needs. All are protected by the present invention.

[0011] Furthermore, the dosage form of the composition includes at least one of water, emulsion, essence, cream, ointment, gel, mask, and powder. In a specific embodiment of the present invention, the dosage form of the composition is a freeze-dried mask.

[0012] Furthermore, the composition can effectively inhibit the activity of tyrosinase and promote the regeneration of collagen and elastin, thereby having a firming and anti-wrinkle effect and being able to significantly resist aging.

[0013] The second aspect of the present invention provides the use of the composition in any of the following aspects: A1: Application in the preparation of anti-aging products; A2: Application in the preparation of firming and anti-wrinkle products; A3: Application in the preparation of products for promoting collagen and / or elastin regeneration; A4: Application in the preparation of products that inhibit tyrosinase activity.

[0014] Furthermore, the anti-aging product, firming and anti-wrinkle product, product for promoting collagen and / or elastin regeneration or product for inhibiting tyrosinase activity includes cosmetics.

[0015] Furthermore, the composition of the present invention is an anti-aging freeze-dried facial mask, which comprises: 0.1 to 15.0 parts by weight of recombinant human albumin; 0.1 to 10.0 parts by weight of wine lees polysaccharide extract; 0.1 to 5.0 parts by weight of red raspberry extract; 5.0 to 10.0 parts by weight of moisturizer; 0.1 to 0.8 parts by weight of thickener; 0.3 to 1.0 parts by weight of moisturizer; and 0.01 to 0.20 parts by weight of stabilizer.

[0016] In a specific embodiment of the present invention, the anti-aging freeze-dried facial mask comprises: 0.1 to 15.0 parts by weight of recombinant human albumin; 0.1 to 10.0 parts by weight of wine lees polysaccharide extract; 0.1 to 5.0 parts by weight of red raspberry extract; 5.0 to 10.0 parts by weight of glycerol; 0.1 to 0.8 parts by weight of carbomer; 0.3 to 1.0 parts by weight of ethylhexyl palmitate; and 0.01 to 0.20 parts by weight of polysorbate 80.

[0017] The third aspect of the present invention provides a method for preparing the composition, the method comprising the following steps: S1: Preparation of wine lees polysaccharide extract: sterilize wine lees, dry, crush, and pass through a 10-mesh sieve to obtain a crude extract powder of wine lees polysaccharide, extract wine lees polysaccharide by water extraction and alcohol precipitation, soak the crude extract with deionized water in a mass ratio of 1:(3-7) and then extract, concentrate the extract, and use an ethanol solution with a concentration of 85% (v / v) or more to precipitate, centrifuge, suspend with purified water, and then spray dry to obtain a wine lees polysaccharide extract. Preferably, the extraction step can be repeated 2-5 times, and the filtrates of multiple extractions are combined and concentrated; S2: Preparation of red raspberry extract: crush the dried red raspberries, pass through a 10-mesh sieve to obtain powder, soak in 75%-80% (v / v) ethanol for 6-8 hours, remove the ethanol by distillation, and then suspend in deionized water and spray dry to obtain red raspberry extract spray-dried powder; S3: Add spray-dried powder of wine sump polysaccharide extract and spray-dried powder of red raspberry extract in certain mass fractions to aqueous solution of recombinant human albumin or salt buffer, and use a high-pressure homogenizer at 500-1000 bar for 5-15 minutes at a temperature of 35°C-60°C to obtain a mixed solution of the anti-aging composition.

[0018] Furthermore, the spray drying conditions in steps S1 and S2 are: air inlet temperature: 120-160° C.; atomization pressure: 0.1-1.0 MPa; feed rate: 200-800 ml / h.

[0019] Furthermore, in step S1, the method for sterilizing the wine lees is high pressure sterilization at a temperature of 115-125° C. for 15-30 min.

[0020] Furthermore, the specific steps of centrifuging the wine lees polysaccharide extract in step S1 are: 5000-8000 r / min, centrifugation for 15-30 min.

[0021] Furthermore, the method further comprises the step of adding additives acceptable in the field of cosmetics to the anti-aging composition mixed solution obtained in step S3 and mixing the mixture.

[0022] In a specific embodiment of the present invention, the method for preparing the recombinant albumin anti-aging composition comprises the following steps: (1) The wine lees polysaccharide extract is prepared according to the following method: sterilize the wine lees under high pressure (121°C, 15-30min), dry the sterilized wine lees, dry and crush the dried wine lees, pass through a 10-mesh sieve to obtain a powder, extract with deionized water, soak for 1h at a solid-liquid ratio of 1:5, reflux extract at water bath temperature for 15min, extract 3 times, combine the filtrate and concentrate, add 3 times the amount of 95% ethanol to the concentrate and place it in a refrigerator for 24h, centrifuge the alcohol-precipitated polysaccharide precipitate at 5000-8000r / min for 15-30min, suspend the precipitate in purified water, and spray dry the suspended solution to obtain the polysaccharide extract; (2) The red raspberry extract is prepared by the following method: dried red raspberries are crushed, passed through a 10-mesh sieve to obtain a powder, soaked in 75% to 80% (v / v) ethanol for 6 to 8 hours, distilled to remove the ethanol, then suspended in deionized water, spray-dried, and a red raspberry extract spray-dried powder is obtained; (3) Inlet air temperature for spray drying: 120-160°C, preferably 150°C; atomization pressure: 0.1-1.0 MPa, preferably 0.5 MPa; feed rate: 200-800 ml / h, preferably 450 ml / h; (4) Take an appropriate amount of deionized water, add the spray-dried powder of wine lees polysaccharide extract and the spray-dried powder of red raspberry extract to the recombinant human albumin, and use a high-pressure homogenizer at 500-1000 bar for 5-15 minutes at a temperature of 35°C-60°C to obtain a composite solution.

[0023] The beneficial effects of the present invention include but are not limited to: 1. In the composition of the present invention, recombinant human albumin, wine lees polysaccharide extract, and red raspberry extract act synergistically to inhibit the activity of tyrosinase, promote the regeneration of collagen and elastin, have a firming and anti-wrinkle effect, and can effectively resist aging. In addition, the composition of the present invention has simple ingredients, good biocompatibility, no obvious irritation and sensitization to the human body, and can be widely used in the preparation of anti-aging products.

[0024] 2. The recombinant human albumin in the composition of the present invention effectively improves the allergenicity of wine lees polysaccharide extract and red raspberry extract, and expands the application of wine lees polysaccharide extract and red raspberry extract in the field of cosmetics.

[0025] 3. The preparation method of the composition of the present invention has a simple production process and has a high application prospect. DETAILED DESCRIPTION

[0026] The present invention is described in detail below with reference to examples, but the present invention is not limited to these examples. Unless otherwise specified, the raw materials and catalysts in the examples of the present invention are purchased through commercial channels.

[0027] Definition: Recombinant human albumin refers to albumin prepared using recombinant DNA technology for treatment and prevention.

[0028] Source of materials: Recombinant human albumin: Tonghua Anrui Special Biopharmaceutical Co., Ltd., catalog number: ART101S; Grape lees: provided by Tonghua Wine Co., Ltd. Red raspberry: provided by Tonghua Huinan raspberry industry base, equipment: Water bath (HH.S21-8, Shanghai Boxun Medical Biological Instrument Co., Ltd.); Multifunctional fluorescence microplate reader (SPMax 3500FL, Shanghai Shanpu Biotechnology Co., Ltd.); Analytical balance (BSA224S, Sartorius Scientific Instruments Beijing Co., Ltd.); Tyrosinase: (Lot No.: T885829-100KU, Shanghai MacLean Biochemical Technology Co., Ltd.); Professional skin tester: (Model: CBS-805, Taiwan CBS Electronics Co., Ltd., China) Hydrogel mask base fabric (Batch No.: 20231102, Shanghai Fulei New Material Technology Co., Ltd.); pH meter (PB-10, Sartorius Scientific Instruments Beijing Co., Ltd.); Unless otherwise specified, other reagents and instruments used were commercially available.

[0029] Example 1: Preparation of an anti-aging composition containing recombinant human albumin comprising the following components in parts by mass: The effective ingredients are shown in Table 1.

[0030] Table 1 Ingredients of various anti-aging freeze-dried facial masks containing recombinant human albumin

[0031] Table 2 Information of control group

[0032] The specific steps for preparing the recombinant human albumin anti-aging composition are as follows: (1) The wine lees polysaccharide extract is prepared according to the following method: the wine lees are sterilized by high pressure (121°C, 15-30min), the sterilized wine lees are dried, the dried wine lees are dried and then crushed, and passed through a 10-mesh sieve to obtain a powder, which is extracted with deionized water, soaked for 1h at a solid-liquid ratio of 1:5, refluxed for 15min at a water bath temperature, extracted 3 times, the filtrate is combined and concentrated, 3 times the amount of 95% ethanol is added to the concentrate, and the concentrate is placed in a refrigerator for 24h, the alcohol-precipitated polysaccharide precipitate is centrifuged at 5000-8000r / min for 15-30min, the precipitate is suspended in purified water, and the suspended solution is spray-dried to obtain the polysaccharide extract.

[0033] (2) The red raspberry extract is prepared by the following method: dried red raspberries are crushed, passed through a 10-mesh sieve to obtain a powder, soaked in 75% to 80% (v / v) ethanol for 6 to 8 hours, distilled to remove the ethanol, then suspended in deionized water, spray-dried, and a red raspberry extract spray-dried powder is obtained; (3) Inlet air temperature for spray drying: 120-160°C, preferably 150°C; atomization pressure: 0.1-1.0 MPa, preferably 0.5 MPa; feed rate: 200-800 ml / h, preferably 450 ml / h.

[0034] (4) Take an appropriate amount of deionized water, add the spray-dried powder of wine lees polysaccharide extract and the spray-dried powder of red raspberry extract to the recombinant human albumin, and use a high-pressure homogenizer at 500-1000 bar for 5-15 minutes at a temperature of 35°C-60°C to obtain a composite solution.

[0035] Test Example 1: Tyrosinase Activity Inhibition Test 1 ml of the combined solution, 0.9 ml of Tris-HCl buffer solution with a pH of 7.6 and 1 ml of tyrosine with a mass fraction of 0.03% were placed in a test tube, placed in a constant temperature water bath at 37°C for 10 min, 0.1 ml of tyrosinase aqueous solution (1070 U / ml) was added, the solution was mixed evenly, placed in a constant temperature water bath at 37°C for 30 min, and its absorbance A at 475 nm was measured using an ultraviolet spectrophotometer. 供试品 .

[0036] Take 1 ml of Tris buffer, 0.9 ml of Tris-HCl buffer (pH 7.6) and 1 ml of 0.03% tyrosine in a test tube, place it in a constant temperature water bath at 37°C for 10 min, add 0.1 ml of tyrosinase aqueous solution (1070 U / ml), mix the solution evenly, place it in a constant temperature water bath at 37°C for 30 min, and measure its absorbance A at 475 nm using a UV spectrophotometer. 阴性 .

[0037] 1 ml of the combined solution, 0.9 ml of Tris hydrochloric acid buffer solution with a pH value of 7.6, and 1 ml of tyrosine with a mass fraction of 0.03% were placed in a test tube, and placed in a constant temperature water bath at 37°C for 10 min. 0.1 ml of Tris hydrochloric acid buffer solution with a pH value of 7.6 was added, and the solution was mixed evenly, and then placed in a constant temperature water bath at 37°C for 30 min. The absorbance A at 475 nm was measured using an ultraviolet spectrophotometer. 空白 . Calculate the inhibition rate of the test sample on tyrosinase. The data results are shown in Table 3.

[0038] Inhibition rate (%) = [(A 阴性 -A 空白 )-(A 供试品 -A 空白 ) / (A 阴性 -A 空白 )×100% Table 3 Tyrosinase activity inhibition test results

[0039] As can be seen from the data in Table 3, the experimental results of the recombinant human albumin anti-aging composition inhibiting tyrosinase show that the tyrosinase inhibition rate is above 69%, indicating that the prepared recombinant human albumin anti-aging composition has an inhibitory effect on tyrosinase.

[0040] The experimental data of the experimental groups (groups 14-16) that only added recombinant human albumin, wine lees polysaccharide extract and red raspberry extract were the lowest in tyrosinase inhibition rate compared with the experimental groups (groups 1-13, 17-18), indicating that the single component has the weakest tyrosinase inhibition ability; at the same time, the data results of the experimental groups (11-13) with the combination of two functional ingredients were much lower than those of groups 1-7, indicating that the anti-aging facial mask liquid can only maintain a strong enzyme inhibition rate under the premise of the coexistence of three groups of active ingredients; the optimal formula ratio, The experimental results of the experimental groups (Groups 1-7) were higher than those of the soy protein group, bovine serum albumin group and egg serum albumin group, indicating that the inhibitory effect of recombinant human albumin on tyrosinase was better than that of serum albumin from plants and animals. Compared with other compatible groups (Groups 8-18), the tyrosinase inhibition rate was >93%. At the same time, compared with the blank control group (Group 19), Group 4 had significant differences, indicating that the recombinant human albumin anti-aging composition prepared by this method has significant inhibitory effect on tyrosinase and can play an anti-aging role.

[0041] Test Example 2: Human skin irritation test According to the "Technical Specifications for Safety of Cosmetics" (2015 edition), human patch tests were conducted. 45 males and 45 females aged 20 to 30 were selected as subjects (without skin diseases and iatrogenic skin sensitivity) and randomly divided into nine groups, with 5 males and 5 females in each group for patch tests. Patch tests were conducted on the inner forearms and behind the ears of the subjects. The test patch tester was removed after 30 minutes of the test, and the compositions prepared by the experimental groups (groups 1, 4, 7, 11, 12, 13, 14, 15, and 16) were selected for testing. The test patch tester was removed after 0.5, 12, and 24 hours of the test, and the skin reaction was observed after the test was completed, and the evaluation was performed with reference to Table 4.

[0042] Table 4 Evaluation criteria for skin patch test

[0043] The experimental results showed that the patch test on the subjects showed that 4 of the subjects using group 12 had weak positive reactions at 24 hours, and the symptoms disappeared after stopping use; 2 of the subjects using groups 14 and 15 had weak positive reactions at 24 hours, and the symptoms disappeared after stopping use; 1 subject using groups 11 and 13 had a suspected erythema reaction; the patch test results of the subjects using groups 1, 4, and 7 were all negative. The above experimental results show that the use of groups 12, 14, and 15 may produce sensitization effects, especially wine lees. The probability of sensitization of polysaccharide extract and red raspberry extract is higher, indicating that the subject's immune system has produced a certain degree of immune response to the test substance, but the degree of reaction is relatively weak. After stopping use, the corresponding symptoms disappear immediately; one person in group 11 had a suspected erythema reaction; the subjects who used the combination of recombinant human albumin, red raspberry extract and wine lees polysaccharide extract all showed negative reactions. According to the evaluation system of the patch test, the negative results show that under the current experimental conditions, the skin of the subjects in these groups did not show detectable immune or inflammatory reactions to the test substances they contacted. The negative patch test results of groups 1, 4, and 7 were verified, indicating that the designed composition may have changed the characteristics of wine lees polysaccharide extract and red raspberry extract that may cause allergies. The encapsulation effect of recombinant human albumin or the specific binding with certain receptors of recombinant human albumin may reduce the sensitization of wine lees extract and red raspberry extract.

[0044] The composition prepared based on the invention patent has no obvious irritation or sensitization to human skin in the actual application scenario simulated in this experiment. From the perspective of safety assessment indicators, the composition has good safety performance.

[0045] Test Example 3: Anti-wrinkle and firming test Elastin and collagen have the function of preventing skin aging and promoting regeneration, making the skin elastic and firm. CBS-805 professional skin tester is used to test elastin and collagen.

[0046] According to the "Technical Specifications for Safety of Cosmetics" (2015 edition), the subjects were screened and 90 Chinese women aged 35 to 65 were selected and randomly divided into three groups. The compositions prepared by the experimental groups (groups 1, 4 and 7) were selected. The subjects used the composition once a day after cleansing their faces and evenly applied the corresponding composition to the cheeks of the face. Other anti-wrinkle products were stopped during the experiment. Skin tests were conducted at week 0, 2, 4 and 8. Before the test, the facial skin was cleaned with water and the subjects were seated in a constant temperature and humidity room for 30 minutes after cleaning. After 30 minutes, the subjects who participated in the experiment were tested for elastin and collagen, and the control was selected as the control before and after. The elastin and collagen test data of all subjects before using the product were used as the control group indicators, and the indicators of the test subjects after using the product for 2 weeks, 4 weeks and 8 weeks were used as the experimental group indicators. The experimental results are shown in Tables 5 and 6.

[0047] Table 5 Elastin test results

[0048] Note: Compared with week 0, P≤0.05, * has significant difference.

[0049] According to the indicators of the CBS-805 professional skin tester, the elastin value of the subjects was detected. The higher the value, the tighter the skin. The value detected by this instrument was in the range of 59~70, and the skin condition was firm. According to the test results in Table 5: the elastin content of the subjects using groups 1, 4 and 7 increased significantly after 2 weeks, 4 weeks and 8 weeks of testing, among which the effect of group 4 was significantly higher than that of the subjects using groups 1 and 7, indicating that at different time points, group 4 had a more significant effect on improving the elastin value. However, although the elastin of the subjects in groups 11-16 using a single component or a combination of two components also increased, the effect was significantly lower than that of groups 1, 4 and 7 using the synergistic combination of the three, indicating that the specific synergistic effect of the recombinant human albumin, wine lees extract and red raspberry extract in the anti-aging composition prepared by this method can significantly promote the regeneration of elastin.

[0050] Table 6 Collagen test results

[0051] Note: Compared with week 0, P≤0.05, * has significant difference.

[0052] According to the CBS-805 professional skin tester index, when the test value is in the range of 25-50, the collagen fibers are missing, when it is 50-65, the collagen fibers are sparse, and when it is greater than 65, the collagen fibers are normal. According to the test results in Table 6: In terms of collagen index, the collagen content of the subjects using groups 1, 4 and 7 increased significantly after 2 weeks, 4 weeks and 8 weeks of testing, among which the collagen content of the subjects in group 4 was significantly higher than that of the subjects using groups 1 and 7, indicating that the combination method used in group 4 has a more prominent effect in increasing the collagen content. However, although the elastin of the subjects in groups 11-16 using a single component or a combination of two components also increased, the effect was significantly lower than that of groups 1, 4 and 7 using the synergistic combination of the three, indicating that the specific synergistic effect of the recombinant human albumin, wine lees extract and red raspberry extract in the anti-aging composition prepared by this method can effectively increase the collagen content in the skin, thereby improving the collagen fiber state of the skin and helping to maintain the normal structure and function of the skin.

[0053] In summary, the anti-aging composition prepared by this method can significantly promote the regeneration of collagen and elastin, increase the content of collagen and elastin, and has anti-wrinkle and firming effects.

[0054] Example 2: Preparation of an anti-aging freeze-dried facial mask containing recombinant human albumin comprising the following components in parts by mass: The effective ingredients are shown in Table 7.

[0055] Table 7 Ingredients of various anti-aging freeze-dried facial masks containing recombinant human albumin

[0056] The specific steps of preparing an anti-aging freeze-dried facial mask containing recombinant human albumin are as follows: (1) The wine lees polysaccharide extract is prepared according to the following method: the wine lees are sterilized by high pressure (121°C, 15-30min), the sterilized wine lees are dried, the dried wine lees are dried and then crushed, and passed through a 10-mesh sieve to obtain a powder, which is extracted with deionized water, soaked for 1h at a solid-liquid ratio of 1:5, refluxed for 15min at a water bath temperature, extracted 3 times, the filtrate is combined and concentrated, 3 times the amount of 95% ethanol is added to the concentrate, and the concentrate is placed in a refrigerator for 24h, the alcohol-precipitated polysaccharide precipitate is centrifuged at 5000-8000r / min for 15-30min, the precipitate is suspended in purified water, and the suspended solution is spray-dried to obtain the polysaccharide extract.

[0057] (2) The red raspberry extract is prepared by the following method: dried red raspberries are crushed, passed through a 10-mesh sieve to obtain a powder, soaked in 75% to 80% (v / v) ethanol for 6 to 8 hours, distilled to remove the ethanol, then suspended in deionized water, spray-dried, and a red raspberry extract spray-dried powder is obtained; (3) Inlet air temperature for spray drying: 120-160°C, preferably 150°C; atomization pressure: 0.1-1.0 MPa, preferably 0.5 MPa; feed rate: 200-800 ml / h, preferably 450 ml / h.

[0058] (4) Take an appropriate amount of deionized water, add the moisturizer, stabilizer and thickener into the beaker in turn, use a digital constant temperature magnetic stirrer to stir until uniform and free of insoluble matter, then heat to about 80°C, maintain for 20 minutes, and then cool to below 40°C as phase A. Take the moisturizer and stir at 35-80rpm, temperature of 55°C-85°C, time of 15-45 minutes to obtain phase B. Add phase B, phase A, spray-dried powder of wine lees polysaccharide extract and spray-dried powder of red raspberry extract to recombinant human albumin, use a high-pressure homogenizer at 500-1500 bar, high-pressure homogenization for 30 minutes, and temperature of 25°C-50°C to obtain a facial mask solution.

[0059] (5) Preparation of anti-aging freeze-dried mask: The mask solution is evenly dispersed on the hydrogel membrane cloth and freeze-dried by freeze-drying flash release technology. The freeze-drying procedure is to perform 2 to 4 sequential addition sublimation steps under the pre-freezing condition of -196°C and vacuum degree of 195 to 205 Pa.

[0060] Test Example 4: Freeze-dried mask tyrosinase activity inhibition test The supernatants of each group of facial masks prepared in Example 2 were used to replace the experimental steps in Test Example 1. The results are shown in Table 8.

[0061] Table 8 Tyrosinase activity inhibition test results

[0062] Note: Compared with group 8, P≤0.05, * has significant difference.

[0063] It can be seen from the data in Table 8 that the tyrosinase activity inhibition rate of the freeze-dried facial masks in each group is >93%. Compared with the results in Table 3, the tyrosinase activity inhibition rate of the freeze-dried facial mask in Example 2 is slightly higher than that of the anti-aging composition in Example 1, indicating that the recombinant human albumin anti-aging freeze-dried facial mask liquid prepared by this method has significant inhibitory effect on tyrosinase and can play an anti-aging role.

[0064] Test Example 5: Freeze-dried facial mask hygiene index determination experiment and heavy metal determination According to the microbiological testing method of the "Safety Technical Specifications for Cosmetics" (2015 edition), the total colony count, heat-resistant Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, mold and yeast in a recombinant human albumin mask liquid (groups 1 to 7) prepared in Example 2 were tested and heavy metals were determined. The microbiological index data are shown in Table 9, and the heavy metal detection experimental results are shown in Table 10.

[0065] Table 9 Results of microbiological index determination in each group

[0066] Table 10 Determination results of heavy metal indicators in each group

[0067] It can be seen from the data in Tables 9 and 10 that the test results of various indicators in the hygiene of the recombinant human albumin anti-aging freeze-dried facial mask are in compliance with the regulations.

[0068] Test Example 6: Freeze-dried facial mask human skin irritation test The supernatants of groups 1, 4, and 7 of freeze-dried facial masks prepared in Example 2 were used to replace the experimental steps in Test Example 2.

[0069] The experimental results showed that the patch test on the skin of the subjects showed that the patch test results of all 30 subjects were negative, which showed that the facial cream prepared by the invention patent had no obvious irritation and allergenicity to the human body and had good safety performance.

[0070] Test Example 7: Freeze-dried facial mask anti-wrinkle and firming experiment The supernatants of each group of facial masks prepared in Example 2 were used to replace the experimental steps in Test Example 3. The results are shown in Tables 11 and 12.

[0071] Table 11 Elastin test results

[0072] Note: Compared with week 0, P≤0.05, * has significant difference.

[0073] According to the CBS-805 professional skin tester, the elastin value of the subjects was detected. The higher the value, the tighter the skin. The value detected by this instrument was in the range of 59~70, and the skin condition was tight. According to the test results in Table 11: In terms of elastin index, the elastin content of the subjects using groups 1, 4 and 7 increased significantly after 2 weeks, 4 weeks and 8 weeks of testing. Among them, the elastin of the subjects using group 4 was significantly higher than that of the subjects using group 1 and group 7 after 2 weeks, 4 weeks and 8 weeks of testing, indicating that group 4 has a more significant effect on improving the elastin value at different time points. At the same time, the elastin content after using this product for 8 weeks was significantly different from that of 0 weeks. At the same time, compared with the mask without additives, the elastin content was also significantly increased, indicating that the freeze-dried mask prepared by this method can increase the elastin content.

[0074] Table 12 Collagen test results

[0075] Note: Compared with week 0, P≤0.05, * has significant difference.

[0076] According to the CBS-805 professional skin tester index, when the test value is in the range of 25~50, the collagen fibers are missing, when it is 50~65, the collagen fibers are sparse, and when it is greater than 65, the collagen fibers are normal. According to the test results in Table 12: In terms of collagen indicators, the elastin content of the subjects using groups 1, 4 and 7 increased significantly after 2 weeks, 4 weeks and 8 weeks of testing, among which the collagen content of the subjects in group 4 was significantly higher than that of the subjects using groups 1 and 7 after 2 weeks, 4 weeks and 8 weeks of testing, indicating that the combination method used in group 4 has a more prominent effect in increasing the collagen content. At the same time, the elastin content after using this product for 8 weeks is significantly different from that of 0 weeks. At the same time, when this product is compared with a mask without additives, the elastin content of the users of this product is significantly increased. The mask prepared by this method can effectively increase the content of skin elastin, thereby improving skin firmness.

[0077] In summary, the anti-aging freeze-dried mask prepared by this method can significantly promote the regeneration of collagen and elastin, increase the content of collagen and elastin, and has anti-wrinkle and firming effects.

[0078] The above description is only an embodiment of the present invention. The protection scope of the present invention is not limited by these specific embodiments, but is determined by the claims of the present invention. For those skilled in the art, the present invention may have various changes and variations. Any modification, equivalent substitution, improvement, etc. made within the technical ideas and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. An anti-aging composition, characterized in that The composition comprises: The quantity of recombinant human albumin is 0.1~15.0; 0.1-10.0 parts of wine lees polysaccharide extract; The amount of red raspberry extract is 0.1 to 5.0 parts.

2. The composition according to claim 1, characterized in that The composition may further include additives acceptable in the cosmetic field.

3. The composition according to claim 2, characterized in that The additives include at least one of a preservative, an emulsifier, a humectant, a thickener, a pH adjuster, a surfactant, a stabilizer, an antioxidant, and a moisturizer.

4. The composition according to any one of claims 1 to 3, characterized in that The dosage form of the composition includes at least one of water, emulsion, essence, cream, ointment, gel, mask and powder.

5. Use of the composition according to any one of claims 1 to 3 in any of the following aspects: A1: Application in the preparation of anti-aging products; A2: Application in the preparation of firming and anti-wrinkle products; A3: Application in the preparation of products for promoting collagen and / or elastin regeneration; A4: Application in the preparation of products that inhibit tyrosinase activity.

6. The use according to claim 5, characterized in that: The anti-aging product, firming and anti-wrinkle product, product for promoting collagen and / or elastin regeneration or product for inhibiting tyrosinase activity includes cosmetics.

7. An anti-aging freeze-dried facial mask, characterized in that: The freeze-dried facial mask comprises: The composition of claim 1; The mass fraction of moisturizer is 5.0~10.0 parts; The mass fraction of thickener is 0.1~0.8 parts; The mass fraction of the moisturizer is 0.3~1.0 parts; The mass fraction of the stabilizer is 0.01~0.20 parts.

8. A method for preparing the composition according to any one of claims 1 to 3, characterized in that: The method comprises the following steps: S1: Preparation of wine lees polysaccharide extract: sterilize the wine lees, dry, crush, and pass through a 10-mesh sieve to obtain a crude extract powder of wine lees polysaccharide, soak the crude extract in deionized water at a mass ratio of 1:(3-7) and then extract, concentrate the extract, and use an ethanol solution with a concentration of 85% (v / v) or more to precipitate, centrifuge, suspend in purified water, and spray dry to obtain a wine lees polysaccharide extract; S2: Preparation of red raspberry extract: crush the dried red raspberries, pass through a 10-mesh sieve to obtain powder, soak in 75%-80% (v / v) ethanol for 6-8 hours, remove the ethanol by distillation, and then suspend in deionized water and spray dry to obtain red raspberry extract spray-dried powder; S3: Add spray-dried powder of wine sump polysaccharide extract and spray-dried powder of red raspberry extract in certain mass fractions to aqueous solution of recombinant human albumin or salt buffer, and use a high-pressure homogenizer at 500-1000 bar for 5-15 minutes at a temperature of 35°C-60°C to obtain a mixed solution of the anti-aging composition.

9. The method according to claim 8, characterized in that The spray drying conditions in steps S1 and S2 are: air inlet temperature: 120-160° C.; atomization pressure: 0.1-1.0 MPa; feed rate: 200-800 ml / h.

10. The method according to claim 8, characterized in that The method further comprises the step of adding additives acceptable in the field of cosmetics to the anti-aging composition mixed solution obtained in step S3 and mixing the mixture.

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