WO2022218174A1 - 一种面膜液的制备方法及面膜产品和应用 - Google Patents

一种面膜液的制备方法及面膜产品和应用 Download PDF

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WO2022218174A1
WO2022218174A1 PCT/CN2022/084849 CN2022084849W WO2022218174A1 WO 2022218174 A1 WO2022218174 A1 WO 2022218174A1 CN 2022084849 W CN2022084849 W CN 2022084849W WO 2022218174 A1 WO2022218174 A1 WO 2022218174A1
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Prior art keywords
facial mask
hours
barley
liquid
pineapple
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PCT/CN2022/084849
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English (en)
French (fr)
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刘平平
吴迪
安全
霍彤
权强华
王昌涛
石秀芹
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云南白药集团健康产品有限公司
云南白药集团股份有限公司
北轻家化(黄山)科技有限公司
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Publication of WO2022218174A1 publication Critical patent/WO2022218174A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9794—Liliopsida [monocotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/73—Polysaccharides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/59—Mixtures
    • A61K2800/592—Mixtures of compounds complementing their respective functions
    • A61K2800/5922—At least two compounds being classified in the same subclass of A61K8/18
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95

Definitions

  • the invention belongs to the field of cosmetics, and in particular relates to a preparation method of a facial mask liquid, a facial mask product and application.
  • Natural cosmetics are more and more popular nowadays. Natural cosmetics, in addition to natural ingredients, do not add flavors, preservatives and other potentially irritating ingredients.
  • enzymolysis can transform macromolecular components into small molecules, which are accompanied by the production of other active substances such as polysaccharides and polypeptides.
  • Plants contain a variety of enzymes, including protease, cellulase, amylase, esterase, etc.
  • the enzymes contained in plants can be used to hydrolyze macromolecular proteins into oligopeptides, and starch and cellulose into oligosaccharides. , lipolysis into glycerol, fatty acid.
  • the application of these products in cosmetics is more conducive to the skin's absorption of functional ingredients, and finally achieves a certain effect.
  • the object of the present invention is to overcome the defects in the prior art, and provide a preparation method of facial mask liquid, facial mask product and application.
  • ROS reactive oxygen species
  • PBS refers to: Phosphate Buffered Saline.
  • t-test refers to: T-test.
  • MTT Cytotoxicity Test
  • VE Vitamin E
  • VC Vitamin C
  • MFI Mean Fluorescence Intensity
  • a first aspect of the present invention provides a preparation method of a facial mask liquid, the preparation method comprising the following steps:
  • step (1) the ratio of pineapple to water is 1:0.1 ⁇ 20g/mL, preferably 1:0.2 ⁇ 10g/mL, more preferably 1:0.5 ⁇ 5g/mL mL, more preferably 1:1 g/mL.
  • step (2) the oats, highland barley and barley are pulverized and passed through an 80-mesh sieve.
  • step (2) the proportion by weight of each raw material component in the pretreatment solution is:
  • oats Preferably, 1-6 parts of oats, 0.5-3 parts of highland barley, 0.5-3 parts of coix seed, and 100 parts of water;
  • step (2) the heating temperature is 20-50°C, preferably 25-100°C, more preferably 30-90°C, further preferably 65-80°C;
  • the heating time is 0.1 to 5 hours, preferably 0.1 to 2 hours, more preferably 0.5 to 1.5 hours, further preferably 0.5 to 1 hour; and/or
  • the stirring rate is 10-800 r/min, preferably 10-500 r/min, more preferably 50-400 r/min, further preferably 250-400 r/min.
  • step (3) the addition ratio of the pineapple treatment solution obtained in step (1) and the pretreatment solution obtained in step (2) is 1:1 ⁇ 500g/mL, preferably 1:1: 5 to 400 g/mL, more preferably 1:10 to 200 g/mL, still more preferably 1:50 to 200 g/mL.
  • step (3) the heating temperature is 20-150°C, preferably 25-100°C, more preferably 30-90°C, further preferably 30-60°C;
  • the heating time is 0.1-5 hours, preferably 0.5-3 hours, more preferably 0.5-2 hours, further preferably 1-2 hours; and/or
  • the stirring rate is 10-800 r/min, preferably 10-500 r/min, more preferably 20-300 r/min, and most preferably 100 r/min.
  • step (4) the ratio of Tremella polysaccharide to the enzymatic hydrolysis treatment solution obtained in step (3) is 1:1 ⁇ 2000g/mL, preferably 1:100 ⁇ 1500g/mL; More preferably 1:100-1000 g/mL, most preferably 1:100-500 g/mL; and/or
  • the homogenization time is 1 to 60 minutes, preferably 1 to 30 minutes, more preferably 1 to 20 minutes, and further preferably 5 to 10 minutes.
  • the sterilization temperature is 80-100°C, more preferably 80°C; and/or the sterilization time is 1-5 hours, more preferably 1 hour.
  • the second aspect of the present invention provides the facial mask liquid prepared according to the method of the first aspect.
  • a third aspect of the present invention provides a facial mask or a face-sticking product, which includes the facial mask liquid described in the second aspect.
  • the fourth aspect of the present invention provides the application of the facial mask liquid described in the second aspect in the preparation of an antioxidant product.
  • the antioxidant has the ability to significantly increase the elastin content of fibroblasts.
  • a fifth aspect of the present invention provides a method for improving skin antioxidant capacity, the method comprising: administering to a subject in need:
  • the invention provides a preparation method of facial mask liquid, the method comprises:
  • the ratio of the oats to water is 1:1 ⁇ 300g/mL, preferably 1:5 ⁇ 200g/mL, more preferably 1:20 ⁇ 150g/mL, most preferably 1:80g/mL mL;
  • the ratio of the barley to water is 1:1-500g/mL, preferably 1:10-300g/mL, more preferably 1:30-200g/mL, most preferably 1:100g/mL;
  • the ratio of coix seed and water is 1:1 ⁇ 200g/mL, preferably 1:30 ⁇ 150g/mL, more preferably 1:50 ⁇ 150g/mL, most preferably 1:100g/mL;
  • the ratio of the pineapple extract to the pretreatment solution in (2) is 1:1 to 500 g/mL, preferably 1:5 to 400 g/mL, more preferably 1:10 to 200 g/mL, and most preferably 1:1 to 200 g/mL. 50g/mL;
  • the heating temperature is 1-150°C, preferably 1-100°C, more preferably 10-90°C, most preferably 70°C;
  • the heating time is 0.1 to 5 hours, preferably 0.1 to 2 hours, more preferably 0.5 to 1.5 hours, and most preferably 1 hour
  • the stirring rate is 10-800 r/min, preferably 10-500 r/min, more preferably 50-400 r/min, and most preferably 300 r/min.
  • the heating temperature is 1-150°C, preferably 1-100°C, more preferably 20-90°C, most preferably 35°C;
  • the heating time is 0.1 to 5 hours, preferably 0.5 to 3 hours, more preferably 0.5 to 2 hours, and most preferably 1.5 hours
  • the stirring rate is 10-800 r/min, preferably 10-500 r/min, more preferably 20-300 r/min, and most preferably 100 r/min.
  • the amount of Tremella polysaccharide added is 1:1-2000, preferably 1:100-1500; more preferably 1:200-1000, most preferably 1:500;
  • the homogenization speed is 1000 ⁇ 20000r/min, preferably 2000 ⁇ 15000r/min, more preferably 2500 ⁇ 10000r/min; the best is 6000r/min;
  • the homogenization time is 1 to 60 minutes, preferably 1 to 30 minutes, more preferably 1 to 20 minutes, and most preferably 10 minutes.
  • the facial mask liquid of the present invention does not add any potentially irritating ingredients such as essence, preservatives, etc., in addition to the natural ingredients in the formula. Tests have proved that the facial mask liquid of the present invention has a promoting effect on the proliferation of fibroblasts, and has a certain transdermal absorption effect.
  • Figure 1 shows the HPLC profile of sample 1 at 220 nm in Example 1.
  • FIG. 2 shows the HPLC profile of sample 2 at 220 nm in Example 1.
  • FIG. 3 shows the HPLC profile of sample 3 at 220 nm in Example 1.
  • FIG. 4 shows the HPLC profile of the blank group at 220 nm in Example 1.
  • FIG. 5 shows the toxicity results of the facial mask liquid of the present invention with different concentrations on fibroblasts in Test Example 1.
  • FIG. 6 shows the transdermal absorption results of the mask liquid of the present invention in Test Example 2.
  • FIG. 7 shows the test results of the effect of the facial mask liquid of the present invention on the content of collagen in fibroblasts in Test Example 3.
  • FIG. 7 shows the test results of the effect of the facial mask liquid of the present invention on the content of collagen in fibroblasts in Test Example 3.
  • Tremella polysaccharide purchased from Shanghai Huiwen Biotechnology Co., Ltd.;
  • Oatmeal (fried and cooked), purchased from Zhangjiakou Jianjun Oat Food Co., Ltd.;
  • Fibroblasts were purchased from Saibaikang (Shanghai) Biotechnology Co., Ltd.
  • Human elastase ELISA kit purchased from Abcam;
  • PBS purchased from Beijing Soleibo Technology Co., Ltd.
  • Homogenizer purchased from IKA Co., Ltd., model: T25;
  • the medical centrifuge was purchased from Wuxi Ruijiang Analytical Instrument Co., Ltd., model RJ-TGL-10B.
  • pineapple enzyme solution of the present invention can reduce the molecular weight of the polypeptide.
  • the experimental method is as follows:
  • the pineapple is washed, peeled and cut into pieces, mixed with water in a mass ratio of 1:1 and beaten to obtain a pineapple enzyme solution (1).
  • the pineapple enzyme liquid (1) is added to the pretreatment liquid (2) according to different mass ratios.
  • the ratios of the pineapple enzyme solution (1) and the pretreatment solution (2) were 1:200, 1:100, and 1:50, respectively, and the numbers of the obtained mixed solutions were sample 1, sample 2, and sample 3, respectively, and the blank group was no.
  • Add the pretreatment solution (2) of the pineapple enzyme solution was 1:200, 1:100, and 1:50, respectively, and the numbers of the obtained mixed solutions were sample 1, sample 2, and sample 3, respectively, and the blank group was no.
  • Sample preparation use the mobile phase as the solvent to prepare a sample with a concentration of 5 mg/ml, and then filter it with a microporous membrane (0.45 ⁇ m) for injection.
  • Standard sample bovine serum albumin (Mr 67000), B12 (Mr 1335), and oxidized glutathione (Mr 614) were prepared into mixed standards, and the content of each substance was 5mg/ml.
  • Figures 1 to 4 show the HPLC profiles of sample 1, sample 2, sample 3 and blank group at 220 nm in Example 1, respectively.
  • the peak time, area and molecular weight of sample 1, sample 2, sample 3 and blank group are shown in Table 1 below.
  • This example is used to illustrate the preparation method of the facial mask liquid of the present invention.
  • Oats, highland barley, and barley crush the oats, barley, and barley to 80 mesh for use.
  • Pineapple washed, peeled, cut into pieces, mixed with water at 1:1 (g/ml) and beaten to obtain a pineapple treatment liquid, which was used as an enzyme liquid.
  • Tremella polysaccharide added Tremella polysaccharide to the enzymatic hydrolysis treatment solution obtained in step (3) according to the ratio of Tremella polysaccharide to the enzymatic hydrolysis treatment solution obtained in step (3) of 1:100 (g/mL), and homogenizing it with a homogenizer for 5 minutes.
  • the mask liquid was put into a mask bag and sterilized at 80°C for 1 hour.
  • This example is used to illustrate the preparation method of the facial mask liquid of the present invention.
  • Oats, highland barley, and barley crush the oats, barley, and barley to 80 mesh for use.
  • Pineapple Wash, peel, and dice. 1:1 (g/ml) was mixed with water and beaten to obtain a pineapple treatment liquid, which was used as an enzyme liquid.
  • step (3) (4) adding tremella polysaccharide to the enzymatic hydrolysis treatment solution obtained in step (3) according to the ratio of Tremella polysaccharide to the enzymatic hydrolysis treatment solution obtained in step (3) of 1:500 (g/mL), and homogenizing it with a homogenizer for 10 minutes.
  • the mask liquid was put into a mask bag and sterilized at 80°C for 1 hour.
  • This example is used to illustrate the preparation method of the facial mask liquid of the present invention.
  • Oats, highland barley, and barley crush the oats, barley, and barley to 80 mesh for use.
  • Pineapple washed, peeled, cut into pieces, mixed with water at 1:1 (g/ml) and beaten to obtain a pineapple treatment liquid, which was used as an enzyme liquid.
  • Tremella polysaccharide was added to the enzymatic hydrolysis treatment solution obtained in step (3) according to the ratio of Tremella polysaccharide to the enzymatic hydrolysis treatment solution obtained in step (3) at 1:200 (g/mL), and homogenized by a homogenizer for 10 minutes.
  • the mask liquid was put into a mask bag and sterilized at 80°C for 1 hour.
  • This example is used to illustrate the preparation method of the facial mask liquid of the present invention.
  • Oats, highland barley, and barley crush the oats, barley, and barley to 80 mesh for use.
  • Pineapple washed, peeled, cut into pieces, mixed with water at 1:1 (g/ml) and beaten to obtain a pineapple treatment liquid, which was used as an enzyme liquid.
  • Tremella polysaccharide to the enzymolysis treatment solution obtained in step (3) according to the ratio of Tremella polysaccharide to the enzymolysis treatment solution obtained in step (3) at 1:300 (g/mL), and homogenize for 5 minutes with a homogenizer.
  • the mask liquid was put into a mask bag and sterilized at 80°C for 1 hour.
  • This example is used to illustrate the preparation method of the facial mask liquid of the present invention.
  • Oats, highland barley, and barley crush the oats, barley, and barley to 80 mesh for use.
  • Pineapple washed, peeled, cut into pieces, mixed with water at 1:5 (g/ml) and beaten to obtain a pineapple treatment liquid, which was used as an enzyme liquid.
  • the pineapple treatment liquid and the pretreatment liquid in (1) were mixed in a ratio of 1:200 (g/mL), and heated at 70° C. for 2 hours. Stir at 300 rad/min; cool to room temperature, filter to remove impurities, and leave the supernatant (opaque) to obtain an enzymatic hydrolysis treatment solution.
  • Tremella polysaccharide was added to the enzymatic hydrolysis treatment solution obtained in step (3) according to the ratio of Tremella polysaccharide to the enzymatic hydrolysis treatment solution obtained in step (3) 1:1000 (g/mL), and homogenized by a homogenizer for 20 minutes.
  • the mask liquid was put into a mask bag and sterilized at 90°C for 3 hours.
  • This example is used to illustrate the preparation method of the facial mask liquid of the present invention.
  • Oats, highland barley, and barley crush the oats, barley, and barley to 80 mesh for use.
  • Pineapple washed, peeled, cut into pieces, mixed with water at 1:10 (g/ml) and beaten to obtain a pineapple treatment liquid, which was used as an enzyme liquid.
  • Tremella polysaccharide and step (3) gained enzymolysis treatment solution 1:1500 (g/mL) ratio to step (3) gained Enzymolysis treatment solution, add Tremella polysaccharide, homogenizer homogenize 30 minutes.
  • the mask liquid was put into a mask bag and sterilized at 90°C for 2 hours.
  • This example is used to illustrate the preparation method of the facial mask liquid of the present invention.
  • Oats, highland barley, and barley crush the oats, barley, and barley to 80 mesh for use.
  • Pineapple washed, peeled, cut into pieces, mixed with water at 1:20 (g/ml) and beaten to obtain a pineapple treatment liquid, which was used as an enzyme liquid.
  • Tremella polysaccharide added Tremella polysaccharide to the enzymatic hydrolysis treatment solution obtained in step (3) according to the ratio of Tremella polysaccharide to the enzymatic hydrolysis treatment solution obtained in step (3) of 1:2000 (g/mL), and homogenizing it with a homogenizer for 60 minutes.
  • the mask liquid was put into a mask bag and sterilized at 90°C for 3 hours.
  • This test example is used to illustrate the effect of the extraction liquid mask prepared in Example 2 on the proliferation of fibroblasts.
  • Collect logarithmic phase cells adjust the concentration of cell suspension, add 100 ⁇ L to each well (96-well plate), and plate the cells to adjust the density to 5000 cells/well (the edge wells are filled with sterile PBS), 5% CO 2 , Incubate at 37°C until the cells grow to a certain density, change the medium and add the test drug with different concentration gradients, and use the culture medium without the test drug as a control. 5% CO 2 , incubated at 37°C for 24 hours, 20 ⁇ L of MTT solution (5 mg/mL, 0.5% MTT tetrazolium salt) was added to each well, and the culture was continued for 4 hours.
  • MTT solution 5 mg/mL, 0.5% MTT tetrazolium salt
  • the culture medium can be discarded after centrifugation, and the culture medium containing MTT (FM-Fibroblast Medium purchased from ScienCell) can be added after carefully washing with PBS for 2-3 times. The culture was terminated, and the medium in the well was carefully aspirated. 150 ⁇ L of dimethyl sulfoxide was added to each well and shaken at low speed on a shaker for 10 min to fully dissolve the crystals. The absorbance value of each well was measured at the OD value of 490 nm in an enzyme-linked immunosorbent assay.
  • MTT FM-Fibroblast Medium purchased from ScienCell
  • Cell viability (OD value of assay well-OD value of blank control)/(OD value of cell control group-OD value of blank control)*100%.
  • Results The test results of different concentration gradients of the samples were compared. To investigate the toxicity of different concentrations of facial mask liquid to fibroblasts. The results are shown in Figure 5. The experimental results show that the mask liquid has no obvious cytotoxicity, and when the sample concentration is 2.5% and 10%, the cell survival rate is significantly increased, about 140%. It shows that the mask liquid has a promoting effect on the proliferation of fibroblasts.
  • This test example is used to illustrate the transdermal absorption results of the facial mask liquid prepared in Example 3.
  • mice were sacrificed by decapsulation of their cervical vertebrae, their back hair was quickly shaved with a razor, the back skin was peeled off, and subcutaneous fat and blood vessels were removed, rinsed repeatedly with distilled water until clean, and then rinsed with normal saline several times, and placed in a -80°C refrigerator. Refrigerate for later use (use within 5 days).
  • Q is the cumulative permeation amount
  • S is the transdermal diffusion area
  • V is the volume of the receiving chamber of the improved Franz diffusion cell
  • Cn is the concentration of the receiving liquid during the nth sampling
  • Ci is the concentration of the receiving liquid during the i-th sampling
  • 0.5 is the sample size.
  • This test example is used to illustrate the effect of the facial mask liquid prepared in Example 3 on the elastin content of fibroblasts.
  • human elastase ELISA kit (cusabio, 96T) was used to detect the elastin content of fibroblasts.
  • Standard product configuration Take a standard product from the kit and centrifuge at 6000-10000rpm for 30s. Dissolve with 1ml of sample diluent, and use the pipette tip to align the bottom of the cryovial for 5 times to help dissolve, and mix well to obtain the standard S7. Arrange seven 1.5ml EP tubes (S0-S6) in sequence, and add 350 ⁇ l of sample diluent to each. Pipette 350 ⁇ l of standard S7 into the first centrifuge tube (S6), and mix by gently pipetting. Pipette 350 ⁇ l from S6 into the second EP tube (S5) and gently pipet evenly. And so on for doubling dilutions of the standards. S0 bit sample diluent.
  • washing working solution Dilute the concentrated washing solution with deionized water at a ratio of 1:25.
  • Biotin-labeled antibody working solution Dilute the biotin-labeled antibody solution by 1:100 times with the biotin-labeled antibody diluent, take 10 ⁇ l of the biotin-labeled antibody and add 990 ⁇ l of the biotin-labeled antibody diluent, mix gently, 10 Ready in minutes.
  • Horseradish peroxidase-labeled avidin working solution Dilute the horseradish peroxidase-labeled avidin by 1:100 times with horseradish peroxidase-labeled avidin dilution solution, and take 10 ⁇ l of horseradish peroxidase Add 990 ⁇ l of horseradish peroxidase-labeled avidin dilution solution to enzyme-labeled avidin, mix gently, and prepare within 10 minutes.
  • the test results of the effect of facial mask liquid on the content of elastin in fibroblasts are shown in Figure 7.
  • the model group was a cell group that was only irradiated with UVA and no sample was added, the blank group was a cell group that was not irradiated and no sample was added, and the positive control group was a cell group that added a positive control sample after UVA irradiation (add VC to the positive control sample. ; Action concentration: 0.15mg/ml).
  • This test example is used to illustrate the effect of the facial mask liquid prepared in Examples 2 to 8 on the content of reactive oxygen species (ROS) in fibroblasts.
  • ROS reactive oxygen species
  • Seeding Seeding fibroblasts in a 6-well plate at a seeding density of 2.2 ⁇ 10 5 cells/well, and incubating in an incubator (37° C., 5% CO 2 ) overnight.
  • UVA irradiation According to the test grouping, the UVA irradiation group was irradiated with 30J/cm 2 of UVA, the irradiation dose was 18.75mW/cm 2 , and the irradiation time was 26min 40s.
  • results The results of the effect of the facial mask liquid prepared in Examples 2 to 8 on the content of reactive oxygen species (ROS) in fibroblasts are shown in Table 10. It can be seen from the table that UVA irradiation can significantly increase the content of ROS in fibroblasts. content, after the sample prepared by the present invention is processed, ROS is significantly reduced. When the ROS content in the body is too high, it will cause a series of damages to the body. ROS is a commonly used indicator for detecting antioxidant and anti-aging activities. It can be seen from the experimental results that the facial mask liquid prepared by the present invention can significantly reduce the ROS content induced by UVA. , which proves that the mask liquid has certain antioxidant and anti-aging effects.
  • ROS reactive oxygen species

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Abstract

本发明提供了一种面膜液的制备方法，还提供了其面膜产品。本发明的面膜液除了配方成分天然以外，同时不添加香精、防腐剂等具有潜在刺激性成分。试验证明，本发明面膜液对成纤维细胞的增殖具有促进作用，具有一定的透皮吸收效果，本面膜液可显著提高弹性蛋白的含量，并显著降低由UVA诱发的ROS含量，达到抗衰老的功效。

Description

一种面膜液的制备方法及面膜产品和应用
相关申请的交叉引用
本申请要求2021年04月16日提交的第CN202110413456.0号中国发明专利申请的优先权，所述申请以引用的方式整体并入本文。
技术领域
本发明属于化妆品领域，具体涉及一种面膜液的制备方法及面膜产品和应用。
背景技术
天然化妆品如今越来越受到人们的青睐。天然化妆品，除了配方成分天然以外，同时不添加香精、防腐剂等具有潜在刺激性成分。
酶解作为一种广泛应用于食品、药品及化妆品领域的技术，可将大分子成分改造成小分子，并伴随多糖、多肽等其他活性物质产生。植物中含有的酶多种多样，有蛋白酶、纤维素酶、淀粉酶、酯酶等，利用植物中含有的酶可以将大分子蛋白质酶解成寡肽，将淀粉、纤维素酶解成寡糖，将脂肪酶解成甘油、脂肪酸。将这些产物应用在化妆品中，更有利于皮肤吸收功效成分，最终达到一定的功效。
发明内容
因此，本发明的目的在于克服现有技术中的缺陷，提供一种面膜液的制备方法及面膜产品和应用。
在阐述本发明内容之前，定义本文中所使用的术语如下：
术语“ROS”是指：活性氧。
术语“PBS”是指：磷酸缓冲盐溶液。
术语“t-test”是指：T检验。
术语“MTT”是指：细胞毒性实验。
术语“VE”是指：维生素E。
术语“VC”是指：维生素C。
术语“MFI”是指：平均荧光强度。
本发明的第一方面提供了一种面膜液的制备方法，所述制备方法包括以下步骤：
(1)将菠萝清洗去皮切块后与水混合打浆，制得菠萝处理液作为酶液；
(2)将燕麦、青稞、薏仁进行物理破碎后备用，与水混合，加热，得到预处理液；
(3)向步骤(2)所得预处理液中加入步骤(1)所得菠萝处理液，加热搅拌，过滤得上清液，制得酶解处理液；
(4)向步骤(3)所得上清液中加入银耳多糖，均质得所述面膜液。
根据本发明第一方面的方法，其中，步骤(1)中，菠萝与水的比例为1:0.1～20g/mL，优选为1:0.2～10g/mL，更优选为1:0.5～5g/mL，进一步优选为1:1g/mL。
根据本发明第一方面的方法，其中，步骤(2)中，所述燕麦、青稞、薏仁粉碎后过80目筛。
根据本发明第一方面的方法，其中，步骤(2)中，所述预处理液中各原料成分的重量份数配比为：
燕麦1～10份，青稞0.5～5份，薏仁0.5～5份，水100份；
优选地，燕麦1～6份，青稞0.5～3份，薏仁0.5～3份，水100份；
更优选地，燕麦2～5份，青稞1～2份，薏仁1～2份，水100。
根据本发明第一方面的方法，其中，步骤(2)中，所述加热温度为20～50℃，优选为25～100℃，更优选为30～90℃，进一步优选为65～80℃；
所述加热时间为0.1～5小时，优选为0.1～2小时，更优选为0.5～1.5小时，进一步优选为0.5～1小时；和/或
所述搅拌速率为10～800r/min，优选为10～500r/min，更优选为50～400r/min，进一步优选为250～400r/min。
根据本发明第一方面的方法，其中，步骤(3)中，步骤(1)所得菠萝处理液和步骤(2)所得预处理液的加入比例为1:1～500g/mL，优选为1:5～400g/mL，更优选为1:10～200g/mL，进一步优选为1:50～200g/mL。
根据本发明第一方面的方法，其中，步骤(3)中，所述加热温度为20～150℃，优选为25～100℃，更优选为30～90℃，进一步优选为30～60℃；
所述加热时间为0.1～5小时，优选为0.5～3小时，更优选为0.5～2小时，进一步优选为1～2小时；和/或
所述搅拌速率为10～800r/min，优选为10～500r/min，更优选为20～300r/min，最优选为100r/min。
根据本发明第一方面的方法，其中，步骤(4)中，银耳多糖与步骤(3)所得酶解处理液的比例为1:1～2000g/mL，优选为1:100～1500g/mL；更优选为1:100～1000g/mL，最优选为1:100～500g/mL；和/或
所述均质时间为1～60分钟，优选为1～30分钟，更优选为1～20分钟，进一步优选为5～10分钟。
根据本发明第一方面的方法，其中，所述方法进一步包括以下步骤：
(5)对步骤(4)所得面膜液进行灭菌；
优选地，所述灭菌温度为80～100℃，更优选为80℃；和/或所述灭菌时间为1～5小时，更优选为1小时。
本发明的第二方面提供了根据第一方面所述的方法制备的面膜液。
本发明的第三方面提供了一种面膜或面贴产品，所述面膜或面贴产品包括第二方面所述的面膜液。
本发明的第四方面提供了第二方面所述的面膜液在制备用于抗氧化产品中的应用。
根据本发明第四方面的应用，其中，所述抗氧化为具有显著提高成纤细胞弹性蛋白含量的能力。
根据本发明第四方面的应用，其中，所述抗氧化为具有显著降低由UVA诱发的ROS含量的能力。
本发明的第五方面提供了一种提高皮肤抗氧化能力的方法，所述方法包括：对有需要的受试者给予：
第二方面所述的面膜液；或
第三方面所述的面膜或面贴产品。
本发明提供了一种面膜液的制备方法，所述方法包括：
(1)将菠萝与水混合打浆制备预处理液，作为酶液。
(2)将燕麦、青稞、薏仁进行物理破碎后备用，与水混合，加热。
(3)(2)中加入(1)预处理液，加热搅拌制备酶解处理液。
(4)将(3)中提取液过滤除去杂质，加入银耳多糖，均质得所述面膜液。
(5)100℃2h灭菌，无菌条件下灌装。
步骤(2)中，所述燕麦和水的比例为1:1～300g/mL，优选为1:5～200g/mL，更优选为1:20～150g/mL，最优选为1:80g/mL；
所述青稞和水的比例为1:1～500g/mL，优选为1:10～300g/mL，更优选为1:30～200g/mL，最优选为1:100g/mL；
所述薏仁和水的比例为1:1～200g/mL，优选为1:30～150g/mL，更优选为1:50～150g/mL，最优选为1:100g/mL；
所述菠萝提取液和(2)中预处理液的比例为1:1～500g/mL，优选为1:5～400g/mL，更优选为1:10～200g/mL，最优选为1:50g/mL；
所述加热温度为1～150℃，优选为1～100℃，更优选为10～90℃，最优选为70℃；
所述加热时间为0.1～5小时，优选为0.1～2小时，更优选为0.5～1.5小时，最优选为1小时
所述搅拌速率为10～800r/min，优选为10～500r/min，更优选为50～400r/min，最优选为300r/min。
步骤(3)中，所述加热温度为1～150℃，优选为1～100℃，更优选为20～90℃，最优选为35℃；
所述加热时间为0.1～5小时，优选为0.5～3小时，更优选为0.5～2小时，最优选为1.5小时
所述搅拌速率为10～800r/min，优选为10～500r/min，更优选为20～300r/min，最优选为100r/min。
步骤(4)中，银耳多糖的加入量为1:1～2000，优选为1:100～1500；更优选为1:200～1000，最优选为1:500；
所述均质速度为1000～20000r/min，优选为2000～15000r/min，更优选为2500～10000r/min；最优为6000r/min；
均质时间为1～60分钟，优选为1～30分钟，更优选为1～20分钟，最优选为10分钟。
本发明的面膜液可以具有但不限于以下有益效果：
本发明的面膜液除了配方成分天然以外，同时不添加香精、防腐剂等具有潜在刺激性成分。试验证明，本发明面膜液对成纤维细胞的增殖具有促进作用，具有一定的透皮吸收效果，本面膜液可显著提高胶原蛋白的含量，达到抗衰老的功效。
附图的简要说明
以下，结合附图来详细说明本发明的实施方案，其中：
图1示出了实施例1中220nm样品1的HPLC图谱。
图2示出了实施例1中220nm样品2的HPLC图谱。
图3示出了实施例1中220nm样品3的HPLC图谱。
图4示出了实施例1中220nm空白组的HPLC图谱。
图5示出了试验例1中本发明不同浓度的面膜液对成纤维细胞的毒性结果。
图6示出了试验例2中本发明面膜液的透皮吸收结果。
图7示出了试验例3中本发明面膜液对成纤维细胞内胶原蛋白含量的影响测试结果。
实施发明的最佳方式
下面通过具体的实施例进一步说明本发明，但是，应当理解为，这些实施例仅仅是用于更详细具体地说明之用，而不应理解为用于以任何形式限制本发明。实施例中未注明具体技术或条件者，按照本领域内的文献所描述的技术或条件，或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者，均为可通过正规渠道商购获得的常规产品。
本部分对本发明试验中所使用到的材料以及试验方法进行一般性的描述。虽然为实现本发明目的所使用的许多材料和操作方法是本领域公知的，但是本发明仍然在此作尽可能详细描述。本领域技术人员清楚，在上下文中，如果未特别说明，本发明所用材料和操作方法是本领域公知的。
以下实施例中使用的试剂和仪器如下：
材料：
菠萝，购自广东省湛江市红星农场；
银耳多糖，购自上海辉文生物技术股份有限公司；
燕麦(炒制熟)，购自张家口建军燕麦食品有限公司；
青稞、薏仁，购自中国北京同仁堂(集团)有限责任公司；
橄榄油，购自云南誉成隆经贸有限公司；
VE，购自Sigma公司；
VC，购自碧云天生物技术有限公司；
成纤维细胞，购自赛百慷(上海)生物技术股份有限公司。
试剂：
人弹性蛋白酶ELISA试剂盒，购自Abcam公司；
MTT试剂盒，购自ScienCell有限公司；
PBS，购自北京索莱宝科技有限公司。
仪器：
均质机，购自IKA有限公司，型号：T25；
酶联免疫检测仪，购自Thermo Fisher Scientific OY有限公司、型号1510-00662C；
医用离心机，购自无锡市瑞江分析仪器有限公司，型号RJ-TGL-10B。
实施例1
本实施例用于说明本发明的菠萝酶液能够降低多肽分子量。
本实验探究了菠萝作为酶液、燕麦作为底物时，酶液与底物的比例对燕麦多肽分子量的影响。
实验方法为以下步骤：
(1)将菠萝清洗去皮切块后与水按照1：1的质量比混合打浆，制得菠萝酶液(1)。
(2)将燕麦与水按照质量比1：25混合，得到预处理液(2)。
(3)预处理液(2)中按照不同质量比加入菠萝酶液(1)。菠萝酶液(1)与预处理液(2)的比例分别为1：200、1：100、1：50，所得的混合液编号分别为样品1、样品2、样品3，其中空白组为不加入菠萝酶液的预处理液(2)。
(4)将(3)中的混合液于35℃保温1小时，后置于95℃水浴0.5小时使酶灭活。
(5)将(4)中的混合液于5000rpm离心20分钟得到上清液。
(6)利用高效液相色谱法对上清液的多肽分子量进行测定。具体测定方法如下：
(a)色谱条件
色谱柱：TsK gel 2000 SWXL 300mm×7.8mm
流动相：0.05mol/L磷酸盐缓冲液(pH 7)+0.3mol/LNaCl
检测波长：UV 220nm
流速：1ml/min
柱温：25℃
样品制备：以流动相为溶剂配制浓度为5mg/ml的样品，再在用微孔膜(0.45μm)过滤后供进样。
标准样品：将牛血清蛋白(Mr 67000)、B12(Mr 1335)、氧化型谷胱甘肽(Mr 614)配成混标，每中物质含量均为5mg/ml。
(b)标准曲线的测定
将三种标准样品按5mg/ml配制，按照上述色谱条件制作标准曲线，采用最小二乘法，求出直线的回归方程为：y＝-0.3913x+7.4563，其回归系数R 2＝0.9982。
由方程的回归系数R 2＝0.9982可知，混合标样的标准曲线线性关系较好，可以提高计算的精确程度。关系式中X代表洗脱时间，Y代表分子量对数。这样当面膜样品进行HPLC分析时，即可通过洗脱图谱中每种物质洗脱峰的出现时间计算其对应的分子量。
(c)分子量测定结果
图1至图4分别示出了实施例1中220nm样品1、样品2、样品3和空白组的HPLC图谱。样品1、样品2、样品3和空白组的出峰时间、面积和分子量如下表1所示。
表1样品1、样品2、样品3和空白组的出峰时间、面积和分子量
Figure PCTCN2022084849-appb-000001
Figure PCTCN2022084849-appb-000002
实验结果：空白组样品中，分子量为188618.64Da的多肽含量为28.89％，分子量为1994.77Da多肽含量为71.11％。而样品1中，分子量为1742.52Da的多肽含量为91.3％；样品2中，分子量为2032.69Da的多肽含量为99.64％；样品3中，分子量为2043.28Da的多肽含量为100％。结果表明，与空白组相比，加入菠萝酶液所得的上清液中，分子量为2000Da左右的多肽占比增加，尤其在样品3中，菠萝中的蛋白酶能够将空白组中28.89％的188618.64Da多肽分子降解为2043.28Da的多肽分子。由此说明，菠萝蛋白酶能够降解燕麦中的高分子量的多肽成为较低分子量多肽。
实施例2
本实施例用于说明本发明面膜液的制备方法。
1.原料处理(食品级)
燕麦、青稞、薏仁：将燕麦、青稞、薏仁粉碎后过80目备用。
菠萝：清洗、去皮、切块，1:1(g/ml)与水混合后打浆制得菠萝处理液，作为酶液使用。
2.面膜液制备
(1)燕麦、青稞、薏仁和水添加比例如表2所示。
表2预处理液的原料比例
原料 质量份数
水 100
燕麦 5
青稞 2
薏仁 2
(2)65℃加热1小时，250rad/min进行搅拌；冷却至室温。
(3)菠萝处理液与(1)中预处理液按照1：50(g/mL)的比例混合，30℃加热2小时。50rad/min进行搅拌；冷却至室温，过滤除去杂质，留上清液(不透明)，制得酶解处理液。
(4)按照银耳多糖与步骤(3)所得酶解处理液1：100(g/mL)比例向步骤(3)所得酶解处理液中加入银耳多糖，均质机均质5分钟。
3.面膜液灭菌处理
面膜液装入面膜袋，80℃灭菌1小时。
实施例3
本实施例用于说明本发明面膜液的制备方法。
1.原料处理(食品级)
燕麦、青稞、薏仁：将燕麦、青稞、薏仁粉碎后过80目备用。
菠萝：清洗、去皮、切块。1:1(g/ml)与水混合后打浆制得菠萝处理液，作为酶液使用。
2.面膜液制备
(1)燕麦、青稞、薏仁和水添加比例如表3所示。
表3预处理液的原料比例
原料 质量份数
水 100
燕麦 2
青稞 1
薏仁 1
(2)80℃加热0.5小时，400rad/min进行搅拌；冷却至室温。
(3)菠萝处理液与(2)中预处理液按照1：50(g/mL)的比例混合，60℃加热1小时。200rad/min进行搅拌；冷却至室温，过滤除去杂质，留上清液(不透明)，制得酶解处理液。
(4)按照银耳多糖与步骤(3)所得酶解处理液1：500(g/mL)比例向步骤(3)所得酶解处理液中加入银耳多糖，均质机均质10分钟。
3.面膜液灭菌处理
面膜液装入面膜袋，80℃灭菌1小时。
实施例4
本实施例用于说明本发明面膜液的制备方法。
1.原料处理(食品级)
燕麦、青稞、薏仁:将燕麦、青稞、薏仁粉碎后过80目备用。
菠萝:清洗、去皮、切块，1:1(g/ml)与水混合后打浆制得菠萝处理液，作为酶液使用。
2.面膜液的制备
(1)燕麦、青稞、薏仁和水的添加比例如表4所示。
表4预处理液的原料比例
原料 质量份数
水 100
燕麦 3
青稞 1.5
薏仁 1.5
(2)70℃加热1小时，300rad/min进行搅拌；冷却至室温。
(3)菠萝处理液与(1)中预处理液按照1:90(g/mL)的比例混合，55℃加热2小时。100rad/min进行搅拌；冷却至室温，过滤除去杂质，留上清液(不透明)，制得酶解处理液。
(4)按照银耳多糖与步骤(3)所得酶解处理液1:200(g/mL)比例向步骤(3)所得酶解处理液中加入银耳多糖，均质机均质10分钟。
3.面膜液灭菌处理
面膜液装入面膜袋，80℃灭菌1小时。
实施例5
本实施例用于说明本发明面膜液的制备方法。
1.原料处理(食品级)
燕麦、青稞、薏仁:将燕麦、青稞、薏仁粉碎后过80目备用。
菠萝:清洗、去皮、切块，1:1(g/ml)与水混合后打浆制得菠萝处理液，作为酶液使用。
2.面膜液的制备
(1)燕麦、青稞、薏仁和水的添加比例如表5所示。
表5预处理液的原料比例
原料 质量份数
水 100
燕麦 4
青稞 2
薏仁 1.5
(2)80℃加热1小时，400rad/min进行搅拌；冷却至室温。
(3)菠萝处理液与(1)中预处理液按照1:70(g/mL)的比例混合，45℃加热2小时。100rad/min进行搅拌；冷却至室温，过滤除去杂质，留上清液(不透明)，制得酶解处理液。
(4)按照银耳多糖与步骤(3)所得酶解处理液1:300(g/mL)比例向步骤(3)所得酶解处理液中加入银耳多糖，均质机均质5分钟。
3.面膜液灭菌处理
面膜液装入面膜袋，80℃灭菌1小时。
实施例6
本实施例用于说明本发明面膜液的制备方法。
1.原料处理(食品级)
燕麦、青稞、薏仁:将燕麦、青稞、薏仁粉碎后过80目备用。
菠萝:清洗、去皮、切块，1:5(g/ml)与水混合后打浆制得菠萝处理液，作为酶液使用。
2.面膜液的制备
(1)燕麦、青稞、薏仁和水的添加比例如表6所示。
表6预处理液的原料比例
原料 质量份数
水 100
燕麦 6
青稞 3
薏仁 3
(2)90℃加热1小时，400rad/min进行搅拌；冷却至室温。
(3)菠萝处理液与(1)中预处理液按照1:200(g/mL)的比例混合，70℃加热2小时。300rad/min进行搅拌；冷却至室温，过滤除去杂质，留上清液(不透明)，制得酶解处理液。
(4)按照银耳多糖与步骤(3)所得酶解处理液1:1000(g/mL)比例向步骤(3)所得酶解处理液中加入银耳多糖，均质机均质20分钟。
3.面膜液灭菌处理
面膜液装入面膜袋，90℃灭菌3小时。
实施例7
本实施例用于说明本发明面膜液的制备方法。
1.原料处理(食品级)
燕麦、青稞、薏仁:将燕麦、青稞、薏仁粉碎后过80目备用。
菠萝:清洗、去皮、切块，1:10(g/ml)与水混合后打浆制得菠萝处理液，作为酶液使用。
2.面膜液的制备
(1)燕麦、青稞、薏仁和水的添加比例如表7所示。
表7预处理液的原料比例
原料 质量份数
水 100
燕麦 10
青稞 5
薏仁 5
(2)100℃加热0.5小时，500rad/min进行搅拌；冷却至室温。
(3)菠萝处理液与(1)中预处理液按照1:400(g/mL)的比例混合，100℃加热1小时。500rad/min进行搅拌；冷却至室温，过滤除去杂质，留上清液(不透明)，制得酶解处理液。
(4)按照银耳多糖与步骤(3)所得酶解处理液1:1500(g/mL)比例向步骤(3)所得 酶解处理液中加入银耳多糖，均质机均质30分钟。
3.面膜液灭菌处理
面膜液装入面膜袋，90℃灭菌2小时。
实施例8
本实施例用于说明本发明面膜液的制备方法。
1.原料处理(食品级)
燕麦、青稞、薏仁:将燕麦、青稞、薏仁粉碎后过80目备用。菠萝:清洗、去皮、切块，1:20(g/ml)与水混合后打浆制得菠萝处理液，作为酶液使用。
2.面膜液的制备
(1)燕麦、青稞、薏仁和水的添加比例如表8所示。
表8预处理液的原料比例
原料 质量份数
水 100
燕麦 6
青稞 3
薏仁 2
(2)40℃加热4小时，800rad/min进行搅拌；冷却至室温。
(3)菠萝处理液与(1)中预处理液按照1:500(g/mL)的比例混合，100℃加热0.5小时。800rad/min进行搅拌；冷却至室温，过滤除去杂质，留上清液(不透明)，制得酶解处理液。
(4)按照银耳多糖与步骤(3)所得酶解处理液1:2000(g/mL)比例向步骤(3)所得酶解处理液中加入银耳多糖，均质机均质60分钟。
3.面膜液灭菌处理
面膜液装入面膜袋，90℃灭菌3小时。
试验例1
本试验例用于说明实施例2制备提取液面膜的对成纤维细胞的增殖的影响。
成纤维细胞毒性实验(MTT)
收集对数期细胞，调整细胞悬液浓度，每孔加入100μL(96孔板),铺板使待测细胞调密度至5000个/孔，(边缘孔用无菌PBS填充)，5％CO 2，37℃孵育，至细胞生长至一定密度，换液加入不同浓度梯度的待测药品，以不含待测药品的培养液为对照。5％CO 2，37℃孵育24小时，每孔加入20μL MTT溶液(5mg/mL，即0.5％MTT四唑盐)，继续培养4h。使药物与MTT充分反应，可先离心后弃去培养液，小心用PBS冲2-3遍后，再加入含MTT的培养液(FM-Fibroblast Medium购自ScienCell)。终止培养，小心吸去 孔内培养液。每孔加入150μL二甲基亚砜，置摇床上低速振荡10min，使结晶物充分溶解。在酶联免疫检测仪OD值490nm处测量各孔的吸光值。
细胞活率＝(测定孔OD值-空白对照OD值)/(细胞对照组OD值-空白对照OD值)*100％。
结果：对比样品不同浓度梯度的试验结果。考察不同浓度的面膜液对成纤维细胞的毒性。结果如图5所示。实验结果表明该面膜液未检出明显的细胞毒性，且在样品浓度为2.5％、10％时，细胞存活率明显增加，约为140％。说明本面膜液对成纤维细胞的增殖具有促进作用。
试验例2
本试验例用于说明实施例3制备面膜液的透皮吸收结果。
(1)鼠皮的制备
将裸鼠脱颈椎处死，迅速将其背部毛用剃须刀剃净，剥离背部皮肤并除去皮下脂肪，血管，用蒸馏水反复冲洗至净，再用生理盐水冲洗数遍，置-80℃冰箱中冷藏备用(在5天内用完)。
(2)Franz扩散池进行体外透皮吸收实验
实验过程:向体外渗透扩散装置恒温槽中加入适量水。开启电源和恒温槽磁力搅拌，设定恒温槽中水温为37±0.1℃。将制备好的鼠皮用铁夹子固定在两室扩散池之间，向垂直扩散池的接受池内加入5mL接受液，放入体外渗透扩散装置恒温槽中预热，设定接受池搅拌速度为400r/min。向供给池内分别加入供给液，以保鲜膜封住上口。开始记时，当样品渗透0、2、4、6、8、24小时(根据实际样品确定具体时间间隔)，分别取样500μl置具塞离心管中，每次取样的同时向接受池中补充等量的接受液并排除池中的气泡。将测定样品透明质酸钠含量。再根据一下公式计算出累积透过量Q(mg/cm 2)。其中，扩散池底部直径为1.50cm，样品接触面积为1.77cm 2。
Figure PCTCN2022084849-appb-000003
注:Q为累积透过量，S为透皮扩散面积，V为改进Franz扩散池接受室的体积，Cn为第n次取样时接受液的浓度，Ci为第i次取样时接受液的浓度，0.5为取样量。通过以上公式计算后，做出累积量与时间的曲线图。接受液为浓度为0.9％的NaCl溶液(生理盐水)；供给液分别为待检测样品。
结果：以多糖为考察指标，测试结果如图6。在保证待测液总糖含量一致的情况下，在0-24h面膜液作用时间内，多糖透过率随作用时间延长而不断增加。说明本面膜液具有一定的透皮吸收效果。
试验例3
本试验例用于说明实施例3制备面膜液对成纤维细胞弹性蛋白含量的影响。
本实施例中采用人弹性蛋白酶ELISA试剂盒(cusabio，96T)成纤维细胞弹性蛋白含量进行检测。
(1)实验准备：
a.标准品配置：从试剂盒中取一支标准品，于6000-10000rpm离心30s。用1ml样本稀释液溶解，并用枪头对准冻存管底部反复吸打5次以助溶解，充分混匀得到标准品S7。取7只1.5ml EP管(S0-S6)依次排列，各加入350μl样本稀释液。吸取350μl标准品S7到第一个离心管(S6)，轻轻吹打混匀。从S6中吸取350μl到第二个EP管中(S5)，轻轻吹打均匀。以此类推进行标准品的倍比稀释。S0位样本稀释液。
b.洗涤工作液：浓洗涤液按1：25倍用去离子水进行稀释。
c.生物素标记抗体工作液：生物素标记抗体液按1：100倍用生物素标记抗体稀释液进行稀释，取10μl生物素标记抗体加990μl生物素标记抗体稀释液，轻轻混匀，10分钟内配妥。
d.辣根过氧化物酶标记亲和素工作液：辣根过氧化酶标记亲和素按1：100倍用辣根过氧化酶标记亲和素稀释液进行稀释，取10μl辣根过氧化物酶标记亲和素加990μl辣根过氧化物酶标记亲和素稀释液，轻轻混匀，10分钟内配妥。
(2)具体实验操作流程
a.将各种试剂移至室温(18-25℃)平衡至少30min，按前述方法配置试剂，备用。
b.加样：分别设标准孔、待测样品孔。每孔分别加标准品或待测样本100μl，轻轻晃动摇匀，覆上板贴，37℃温育2小时。
c.弃去液体，甩干，不用洗涤。
d.每孔加生物素标记抗体工作液100μl，覆上新的板贴，37℃温育1小时。
e.弃去孔内液体，甩干，洗板3次。每次浸泡2min，200μl每孔，甩干。
f.每孔加辣根过氧化物酶标记亲和素工作液100μl，覆上新的板贴，37℃温育1小时
g.弃去孔内液体，甩干，洗板5次。每次浸泡2分钟，200μl/每孔，甩干。
h.依次每孔加底物溶液90μl，37℃避光显色15-30分钟。
i.依序每孔加终止溶液50μl，终止反应。
j.在反应终止后5min内用酶标仪在450nm波长依序测量各孔的OD值。
k.计算：根据浓度和OD值算出标准曲线和回归方程，用专用软件ELISACalc进行计算，拟合模型选用logistic曲线1。
结论：面膜液对成纤维细胞内弹性蛋白含量的影响测试结果如图7。模型组是只通过UVA照射、未添加样品的细胞组，空白组为未照射且未添加样品的细胞组，阳性对照组 为UVA照射后，添加阳性对照样品的细胞组(阳性对照样品中添加VC；作用浓度为：0.15mg/ml)。
经试验发现UVA照射会严重损伤细胞的弹性蛋白，样品组与模型组、空白组和阳性对照组对比，本面膜液极其显著提高成纤维细胞弹性蛋白的含量，达到抗衰老的作用。
试验例4
本试验例用于说明实施例2～8制备的面膜液对成纤维细胞内活性氧(ROS)含量的影响。
实验方案设计如表9所示
表9实验方案
Figure PCTCN2022084849-appb-000004
具体操作步骤如下：
(1)接种:按2.2×10 5个/孔的接种密度接种成纤维细胞至6孔板，培养箱(37℃、5％CO 2)中孵育过夜。
(2)给药:根据表9测试方案，待6孔板中细胞铺板率达到40％～60％时，进行分组给药，每孔加样2mL，每组设3个复孔。给药完成后将6孔板放置在培养箱(37℃、5％CO 2)中培养24h。
(3)UVA辐照:根据试验分组，对有UVA照射的组别进行30J/cm 2的UVA辐照，辐照剂量为18.75mW/cm 2，辐照时间为26min 40s。
(4)辐照结束后，直接用PBS清洗每孔细胞3次；
(5)流式检测:每孔加入1mL浓度为10μM DCFH-DA探针，培养箱(37℃、5％CO2)中孵育30min；弃掉含DCFH-DA的培养液，用PBS清洗3次，胰酶(0.25％)消化细胞后，PBS清洗细胞1次，加入一定量新鲜PBS，流式检测；
(6)结果分析:汇总各组MFI(平均荧光强度)，用t-test方法进行统计分析。
结果：实施例2～8制备的面膜液对成纤维细胞内活性氧(ROS)含量的影响结果如表10所示，从表中可以看出，UVA的照射会显著增加成纤维细胞体内ROS的含量，本发明制得得样品处理后，ROS显著下降。当体内ROS含量过高时会造成机体的一系列损伤，ROS是检测抗氧化和抗衰老活性常用的指标，从实验结果可以看出，本发明制备的面膜液可以显著降低由UVA诱发的ROS含量，从而证明该面膜液具有一定的抗氧化和抗衰老功效。
表10细胞内ROS含量变化结果
Figure PCTCN2022084849-appb-000005
备注：用t-test方法进行统计分析时，NC组与BC组相比，显著性以#表示，P-value＜0.05表示为#，P-value＜0.01表示为##；PC组、样品组与NC组相比，显著性以*表示，P-value＜0.05表示为*，P-value＜0.01表示为**。
尽管本发明已进行了一定程度的描述，明显地，在不脱离本发明的精神和范围的条件下，可进行各个条件的适当变化。可以理解，本发明不限于所述实施方案，而归于权利要求的范围，其包括所述每个因素的等同替换。

Claims (15)

  1. 一种面膜液的制备方法，其特征在于，所述制备方法包括以下步骤：
    (1)将菠萝清洗去皮切块后与水混合打浆，制得菠萝处理液作为酶液；
    (2)将燕麦、青稞、薏仁进行物理破碎后备用，与水混合，加热，得到预处理液；
    (3)向步骤(2)所得预处理液中加入步骤(1)所得菠萝处理液，加热搅拌，过滤得上清液，制得酶解处理液；
    (4)向步骤(3)所得酶解处理液中加入银耳多糖，均质得所述面膜液。
  2. 根据权利要求1所述的方法，其特征在于，步骤(1)中，菠萝与水的比例为1:0.1～20g/mL，优选为1:0.2～10g/mL，更优选为1:0.5～5g/mL，进一步优选为1:1g/mL。
  3. 根据权利要求1或2所述的方法，其特征在于，步骤(2)中，所述燕麦、青稞、薏仁粉碎后过80目筛。
  4. 根据权利要求1至3中任一项所述的方法，其特征在于，步骤(2)中，所述预处理液中各原料成分的重量份数配比为：
    燕麦1～10份，青稞0.5～5份，薏仁0.5～5份，水100份；
    优选地，燕麦1～6份，青稞0.5～3份，薏仁0.5～3份，水100份；
    更优选地，燕麦2～5份，青稞1～2份，薏仁1～2份，水100份。
  5. 根据权利要求1至4中任一项所述的方法，其特征在于，步骤(2)中，所述加热温度为20～50℃，优选为25～100℃，更优选为30～90℃，进一步优选为65～80℃；
    所述加热时间为0.1～5小时，优选为0.1～2小时，更优选为0.5～1.5小时，进一步优选为0.5～1小时；和/或
    所述搅拌速率为10～800r/min，优选为10～500r/min，更优选为50～400r/min，进一步优选为250～400r/min。
  6. 根据权利要求1至5中任一项所述的方法，其特征在于，步骤(3)中，步骤(1)所得菠萝处理液和步骤(2)所得预处理液的加入比例为1:1～500g/mL，优选为1:5～400g/mL， 更优选为1:10～200g/mL，进一步优选为1:50～200g/mL。
  7. 根据权利要求1至6中任一项所述的方法，其特征在于，步骤(3)中，所述加热温度为20～150℃，优选为25～100℃，更优选为30～90℃，进一步优选为30～60℃；
    所述加热时间为0.1～5小时，优选为0.5～3小时，更优选为0.5～2小时，进一步优选为1～2小时；和/或
    所述搅拌速率为10～800r/min，优选为10～500r/min，更优选为20～300r/min，最优选为100r/min。
  8. 根据权利要求1至7中任一项所述的方法，其特征在于，步骤(4)中，银耳多糖与步骤(3)所得酶解处理液的比例为1:1～2000g/mL，优选为1:100～1500g/mL；更优选为1:100～1000g/mL，最优选为1:100～500g/mL；和/或
    所述均质时间为1～60分钟，优选为1～30分钟，更优选为1～20分钟，进一步优选为5～10分钟。
  9. 根据权利要求1至8中任一项所述的方法，其特征在于，所述方法进一步包括以下步骤：
    (5)对步骤(4)所得面膜液进行灭菌；
    优选地，所述灭菌温度为80～100℃，更优选为80℃；和/或所述灭菌时间为1～5小时，更优选为1小时。
  10. 根据权利要求1至9中任一项所述方法制备的面膜液。
  11. 一种面膜或面贴产品，其特征在于，所述面膜或面贴产品包括根据权利要求10所述的面膜液。
  12. 根据权利要求10所述的面膜液在制备用于抗氧化产品中的应用。
  13. 根据权利要求12所述的应用，其特征在于，所述抗氧化为具有显著提高成纤细胞弹性蛋白含量的能力。
  14. 根据权利要求12所述的应用，其特征在于，所述抗氧化为具有显著降低由UVA诱发的ROS含量的能力。
  15. 一种提高皮肤抗氧化能力的方法，其特征在于，所述方法包括：对有需要的受试者给予：
    根据权利要求10所述的面膜液；或
    根据权利要求11所述的面膜或面贴产品。
PCT/CN2022/084849 2021-04-16 2022-04-01 一种面膜液的制备方法及面膜产品和应用 WO2022218174A1 (zh)

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Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113230166A (zh) * 2021-04-16 2021-08-10 云南白药集团股份有限公司 一种面膜液、其制备方法及应用
CN113274334B (zh) * 2021-04-16 2022-07-22 云南白药集团股份有限公司 具有抗衰老作用的菠萝提取液组合物、其制备方法和应用
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CN113274333B (zh) * 2021-04-16 2022-07-26 云南白药集团股份有限公司 具有抗氧化作用的菠萝提取液组合物、其制备方法和应用

Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2925329A1 (fr) * 2007-12-21 2009-06-26 Vincience Sa Utilisation d'un hydrolysat d'avoine en tant que principe actif activateur de la synthese des aquaporines
CN102247306A (zh) * 2010-05-18 2011-11-23 北京工商大学 一种燕麦提取物及其制备方法与应用
CN104069045A (zh) * 2014-06-24 2014-10-01 樊世菊 一种祛斑防皱面膜
CN104434647A (zh) * 2014-11-17 2015-03-25 上海应用技术学院 一种含有青稞β-葡聚糖的滋养调理润肤液及其制备方法
CN109646318A (zh) * 2019-01-29 2019-04-19 廖敏超 一种美白抗衰老净肤水及其制备方法
CN113197817A (zh) * 2021-04-16 2021-08-03 云南白药集团股份有限公司 一种菠萝提取液组合物的美白应用
CN113197816A (zh) * 2021-04-16 2021-08-03 云南白药集团股份有限公司 一种面膜液的制备方法及其产品
CN113197818A (zh) * 2021-04-16 2021-08-03 云南白药集团股份有限公司 一种面膜液的制备方法及面膜产品
CN113230166A (zh) * 2021-04-16 2021-08-10 云南白药集团股份有限公司 一种面膜液、其制备方法及应用
CN113274334A (zh) * 2021-04-16 2021-08-20 云南白药集团股份有限公司 具有抗衰老作用的菠萝提取液组合物、其制备方法和应用
CN113274333A (zh) * 2021-04-16 2021-08-20 云南白药集团股份有限公司 具有抗氧化作用的菠萝提取液组合物、其制备方法和应用

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2850277B1 (fr) * 2003-01-24 2005-04-08 Lanatech Composition comprenant un extrait d'aphanizomenon flos-aquae, son utilisation et sa preparation
DE102005021805B3 (de) * 2005-05-04 2006-10-05 Lancaster Group Gmbh Kosmetisches Verfahren zur Körpermodellierung mit Sonnenschutz und Modellierungs-Set

Patent Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2925329A1 (fr) * 2007-12-21 2009-06-26 Vincience Sa Utilisation d'un hydrolysat d'avoine en tant que principe actif activateur de la synthese des aquaporines
CN102247306A (zh) * 2010-05-18 2011-11-23 北京工商大学 一种燕麦提取物及其制备方法与应用
CN104069045A (zh) * 2014-06-24 2014-10-01 樊世菊 一种祛斑防皱面膜
CN104434647A (zh) * 2014-11-17 2015-03-25 上海应用技术学院 一种含有青稞β-葡聚糖的滋养调理润肤液及其制备方法
CN109646318A (zh) * 2019-01-29 2019-04-19 廖敏超 一种美白抗衰老净肤水及其制备方法
CN113197817A (zh) * 2021-04-16 2021-08-03 云南白药集团股份有限公司 一种菠萝提取液组合物的美白应用
CN113197816A (zh) * 2021-04-16 2021-08-03 云南白药集团股份有限公司 一种面膜液的制备方法及其产品
CN113197818A (zh) * 2021-04-16 2021-08-03 云南白药集团股份有限公司 一种面膜液的制备方法及面膜产品
CN113230166A (zh) * 2021-04-16 2021-08-10 云南白药集团股份有限公司 一种面膜液、其制备方法及应用
CN113274334A (zh) * 2021-04-16 2021-08-20 云南白药集团股份有限公司 具有抗衰老作用的菠萝提取液组合物、其制备方法和应用
CN113274333A (zh) * 2021-04-16 2021-08-20 云南白药集团股份有限公司 具有抗氧化作用的菠萝提取液组合物、其制备方法和应用

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
FENG BING, WANG CHANGTAO, LEI FANG, DONG YINMAO: "Research on Hydrolysis of Oat Bran Protein Under the Action of Different Proteases", SCIENCE AND TECHNOLOGY OF FOOD INDUSTRY, GAI KAN BIANJIBU , BEIJING, CN, no. 10, 31 May 2007 (2007-05-31), CN , pages 120 - 122, XP055977595, ISSN: 1002-0306, DOI: 10.13386/j.issn1002-0306.2007.10.026 *
WANG SU, CHEN JIWANG, TIAN XIANGDONG, LIU YING: "Enzymatic Modification of Oat Protein Isolate and Product Properties", ZHONGGUO LIANGYOU XUEBAO - CHINESE CEREALS AND OILS ASSOCIATION.JOURNAL, ZHONGGUO LIANGYOU XUEHUI, BEIJING, CN, vol. 25, no. 12, 31 December 2010 (2010-12-31), CN , pages 32 - 36, XP055977599, ISSN: 1003-0174 *

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