WO2022218170A1 - 具有抗衰老作用的菠萝提取液组合物、其制备方法和应用 - Google Patents
具有抗衰老作用的菠萝提取液组合物、其制备方法和应用 Download PDFInfo
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- WO2022218170A1 WO2022218170A1 PCT/CN2022/084836 CN2022084836W WO2022218170A1 WO 2022218170 A1 WO2022218170 A1 WO 2022218170A1 CN 2022084836 W CN2022084836 W CN 2022084836W WO 2022218170 A1 WO2022218170 A1 WO 2022218170A1
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- pineapple
- extract composition
- pineapple extract
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- oil
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Classifications
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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]
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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]
- A61K8/9789—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
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
Definitions
- the invention belongs to the field of cosmetics, and in particular relates to a pineapple extract composition with anti-aging effect, a preparation method and application thereof.
- 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 purpose of the present invention is to overcome the defects in the prior art, and to provide a pineapple extract composition with anti-aging effect, its preparation method and application.
- GSH-Px refers to: glutathione peroxidase.
- TGF ⁇ refers to: Transforming Growth Factor- ⁇ .
- MDA malondialdehyde
- PBS refers to: Phosphate Buffered Saline.
- t-test refers to: T-test.
- MTT Fibroblast toxicity test
- VC Vitamin C
- the first aspect of the present invention provides a pineapple extract composition with anti-aging effect, characterized in that, the pineapple extract composition is prepared by extracting the raw materials of the following components: pineapple, oat, Papaya, Momordica grosvenori, Pueraria lobata, vegetable oil and water; and the pineapple extract composition is prepared by mixing the oat pretreatment solution prepared by oat and water with the mixed pretreatment solution containing pineapple, papaya, and Luo Han Guo, and then adding Pueraria lobata, vegetable oil be made of.
- the pineapple extract liquid composition according to the first aspect of the present invention wherein, the vegetable oil is selected from one or more of the following: olive oil, green thorn fruit oil, macadamia nut oil, peony seed oil, toon flower oil; and/or
- the parts by mass of each component in the pineapple extract composition are: 0.01-5 parts of pineapple, 0.1-10 parts of oats, 0.01-5 parts of papaya, 0.01-5 parts of Luo Han Guo, 0.5-10 parts of pueraria root powder, and 100 parts of water ;
- 0.1-1 part of pineapple 0.5-2 parts of oat, 0.1-0.5 part of papaya, 0.1-0.5 part of Luo Han Guo, 2-5 parts of Pueraria lobata, and 100 parts of water.
- a second aspect of the present invention provides a method for preparing the pineapple extract composition of the first aspect, the method comprising the following steps:
- step (3) the oat pretreatment solution obtained in step (1) and the mixed pretreatment solution obtained in step (2) are mixed, heated and stirred, and the supernatant is left by cooling and filtration;
- step (3) (4) adding pueraria root powder and vegetable oil to the supernatant obtained in step (3), and homogenizing to obtain the pineapple extract composition.
- step (1) the oats are crushed and pass through a 60-100 mesh, preferably a 80-mesh sieve;
- the mass ratio of the oats to water is 1:1-500, preferably 1:5-200, more preferably 1:20-100, most preferably 1:50; and/or
- the standing time is 8 to 60 hours, preferably 12 to 48 hours, and most preferably 24 hours.
- step (2) the mass ratio of the pineapple to water is 1:1-150, preferably 1:10-100, more preferably 1:20-80, and most Preferably it is 1:20 ⁇ 50;
- the mass ratio of the papaya to water is 1:1-200, preferably 1:10-150, more preferably 1:20-100, most preferably 1:50-80;
- the mass ratio of the Luo Han Guo and water is 1:1-200, preferably 1:10-150, more preferably 1:20-100, most preferably 1:50-80.
- step (3) the mass ratio of the mixed pretreatment solution obtained in step (2) to the oat pretreatment solution obtained in step (1) is 1:1 to 100, preferably 1:1: 1 to 50, more preferably 1:1 to 20, and most preferably 1:10.
- step (3) the heating temperature is 20-90°C, preferably 25-85°C, more preferably 30-80°C, further preferably 40-60°C;
- the heating time is 0.5 to 5 hours, preferably 0.5 to 4 hours, more preferably 1 to 3 hours, further preferably 1 to 2 hours; and/or
- the stirring rate is 50-500 r/min, preferably 50-300 r/min, more preferably 50-200 r/min, further preferably 50-150 r/min.
- step (4) the mass ratio of the pueraria root powder to the supernatant obtained in step (3) is 1:10-200, preferably 1:10-100, more preferably 1:20-80, most preferably 1:25-50; and/or
- the homogenization time is 1 to 60 minutes, preferably 5 to 50 minutes, more preferably 5 to 30 minutes, and further preferably 15 to 20 minutes.
- the sterilization treatment method is heat sterilization
- the heat sterilization temperature is 60-100°C, preferably 70-90°C, most preferably 80°C; and/or the heat-sterilization time is 1-5 hours, preferably 1-3 hours , most preferably 2 hours.
- a third aspect of the present invention provides an application of a pineapple extract composition in the preparation of anti-aging products, the pineapple extract composition comprising pineapple, oat, papaya, Luo Han Guo and Pueraria root powder;
- the pineapple extract composition is the pineapple extract composition of the first aspect or the pineapple extract composition prepared according to the method of the second aspect.
- a fourth aspect of the present invention provides a cosmetic product comprising the pineapple extract composition of the first aspect or the pineapple extract composition prepared by the method of the second aspect;
- the dosage form of the cosmetic product is selected from one or more of the following: cream, lotion, water, gel, oil, powder, mud, aerosol, patch, film;
- the cosmetic product is a cream, mask, lotion or essence.
- the cosmetic product according to the fourth aspect of the present invention wherein, when the cosmetic product is a cream, its preparation method comprises the following steps:
- step (c) adding the oil phase mixture prepared in step (b) to the water phase mixture prepared in step (a), and homogenizing to obtain the cosmetic product.
- the water phase ingredients include vegetable oil and an antibacterial agent; and/or in the step (b), the oil phase ingredients include vegetable oil and emulsifier;
- the vegetable oil is selected from one or more of the following: glycerin, green thorn fruit oil, olive oil, peony seed oil, macadamia nut oil, babassu oil, toona japonica oil;
- the antibacterial agent is selected from one or more of the following: hexanediol, methylparaben, propylparaben, phenoxyethanol; and/or
- the emulsifier is selected from one or more of the following: wheat emulsifier, polyglycerol-3 diisostearate, dipolyhydroxystearic acid, and yeoleyl glucoside.
- the ratio of the sterilized pineapple extract composition, vegetable oil and antibacterial agent is 80 ⁇ 95:1 ⁇ 10:0.01 ⁇ 1g /g/g, preferably 89 ⁇ 93:2 ⁇ 5:0.05 ⁇ 0.2g/g/g;
- the ratio of the vegetable oil to the emulsifier is 1 ⁇ 10:2 ⁇ 15g/g, preferably 2 ⁇ 5:5 ⁇ 10g/g; and/or
- the ratio of the oil phase mixture to the water phase mixture is 2-25:0.5-5g/g, preferably 5-10:1-2g/g;
- the heating temperature is 70-120°C, more preferably 70-100°C, further preferably 80-90°C; and /or
- the homogenization time is 15-45 minutes, more preferably 20-40 minutes, and even more preferably 25-35 minutes.
- a fifth aspect of the present invention provides a method for anti-aging, the method comprising: administering to a subject in need:
- the pineapple extract composition of the first aspect is
- the pineapple extract composition prepared according to the method described in the second aspect; or
- the preparation method of the pineapple extract composition of the present invention comprises the following steps:
- the ratio of the oats to water is 1:1 ⁇ 500g/mL, preferably 1:5 ⁇ 300g/mL, more preferably 1:20 ⁇ 200g/mL, most preferably 1:100g/mL mL;
- the ratio of the pineapple to water is 1:1 ⁇ 150g/mL, preferably 1:10 ⁇ 100g/mL, more preferably 1:20 ⁇ 80g/mL, most preferably 1:50g/mL mL;
- the ratio of papaya to water is 1:1 ⁇ 200g/mL, preferably 1:10 ⁇ 150g/mL, more preferably 1:20 ⁇ 100g/mL, most preferably 1:80g/mL;
- the ratio of the Luo Han Guo to water is 1:1 ⁇ 200g/mL, preferably 1:10 ⁇ 150g/mL, more preferably 1:50 ⁇ 100g/mL, most preferably 1:80g/mL;
- the heating temperature is 1-150°C, preferably 10-100°C, more preferably 20-100°C, most preferably 40°C;
- the heating time is 1-10 hours, preferably 1-8 hours, more preferably 1-5 hours, most preferably 4 hours
- the stirring rate is 10-600 r/min, preferably 10-500 r/min, more preferably 10-300 r/min, and most preferably 150 r/min.
- the ratio of the pueraria root powder to the extract is 1:1 ⁇ 100g/mL, preferably 1:5 ⁇ 80g/mL, more preferably 1:10 ⁇ 70g/mL, most preferably 1:1: 50g/mL;
- the amount of vegetable oil added is 1-30%, preferably 1-20%; more preferably 1-10%, most preferably 3%;
- the homogenization speed is 1000 ⁇ 8000r/min, preferably 1000 ⁇ 7000r/min, more preferably 2000 ⁇ 6000r/min; the most optimal is 5000r/min;
- the homogenization time is 1 to 60 minutes, preferably 1 to 50 minutes, more preferably 10 to 40 minutes, and most preferably 15 minutes.
- the present invention provides the application of a pineapple extract composition in the preparation of products with anti-aging effect.
- the inventors have proved through experiments that the pineapple extract composition of the present invention can increase the degradation of intracellular related peroxides and reduce the After UVA irradiation, the content of MDA in cells increases the degradation of related peroxides in cells, thereby reducing the damage of peroxidation to cells and achieving anti-aging effects.
- FIG. 1 shows the effect on glutathione peroxidase (GSH-Px) in fibroblasts in Test Example 1.
- FIG. 1 shows the effect on glutathione peroxidase (GSH-Px) in fibroblasts in Test Example 1.
- FIG. 2 shows the effect of lipid peroxide content in fibroblasts in Test Example 2.
- FIG. 3 shows the hydrogen peroxide standard curve in Test Example 3.
- FIG. 4 shows the effect of catalase enzyme activity in Test Example 3.
- FIG. 5 is a graph showing an example of changes in wrinkles (forehead lines) of the volunteer 1 after using the cream 1 prepared in Example 8 in Test Example 5.
- FIG. 5 is a graph showing an example of changes in wrinkles (forehead lines) of the volunteer 1 after using the cream 1 prepared in Example 8 in Test Example 5.
- FIG. 6 is a graph showing an example of changes in wrinkles (canthus) of Volunteer 2 after using Cream 1 prepared in Example 8 in Test Example 5.
- FIG. 6 is a graph showing an example of changes in wrinkles (canthus) of Volunteer 2 after using Cream 1 prepared in Example 8 in Test Example 5.
- FIG. 7 is a graph showing an example of changes in wrinkles (pockmarks) of Volunteer 1 after using Cream 2 prepared in Example 9 in Test Example 5.
- FIG. 7 is a graph showing an example of changes in wrinkles (pockmarks) of Volunteer 1 after using Cream 2 prepared in Example 9 in Test Example 5.
- FIG. 8 is a graph showing an example of changes in wrinkles (pockmarks) of Volunteer 2 after using Cream 2 prepared in Example 9 in Test Example 5.
- FIG. 8 is a graph showing an example of changes in wrinkles (pockmarks) of Volunteer 2 after using Cream 2 prepared in Example 9 in Test Example 5.
- Figure 9 shows the effect of the sample and control on the skin R0 in Test Example 5, wherein, *, p ⁇ 0.05, indicating that there is a significant difference compared with the background; n.s., p>0.05, indicating that compared with the background , with no significant difference.
- Figure 10 shows the mean value R2 of the difference between the sample and the control group at each time point and the background in Test Example 5 (comparison between samples), wherein, *, p ⁇ 0.05, indicating that compared with the background, there is a significant Sexual difference; n.s., p>0.05, indicating that there is no significant difference compared with the background.
- Oats (fried and cooked) were purchased from Zhangjiakou Jianjun Oat Food Co., Ltd.;
- Pueraria lobata was purchased from Beijing Tongrentang (Group) Co., Ltd., Beijing, China.
- the kit was purchased from Beijing Soleibo Technology Co., Ltd.;
- Homogenizer purchased from IKA Ltd., model T25.
- the medical centrifuge was purchased from Wuxi Ruijiang Analytical Instrument Co., Ltd., model RJ-TGL-10B.
- Skin elasticity test instrument purchased from Beijing Jinhongfan Trading Co., Ltd., model MPA580 multi-probe.
- Facial image analyzer purchased from Beijing Jinhongfan Trading Co., Ltd., model VISIA CR.
- This example is used to illustrate the preparation method of the pineapple extract composition of the present invention.
- Papaya Wash, peel and cut into pieces.
- Oatmeal After crushing, pass through 80 mesh for use.
- the pretreatment solution and the step (1) oat pretreatment solution are mixed according to a ratio of 1:10g/mL, heated at 40°C for 2 hours, and stirred at 50rad/min; cooled to room temperature, filtered to remove impurities, Leave the supernatant (opaque).
- the pineapple extract liquid composition is filled into a mask bag (containing a mask cloth).
- This example is used to illustrate the preparation method of the pineapple extract composition of the present invention.
- Papaya Wash, peel and cut into pieces.
- Oatmeal After crushing, pass through 80 mesh for use.
- the pretreatment solution and the step (1) oat pretreatment solution are mixed in a ratio of 1:10g/mL, heated at 60°C for 1 hour, and stirred at 150rad/min; cooled to room temperature, filtered to remove impurities, Leave the supernatant (opaque).
- the pineapple extract liquid composition is filled into a mask bag (containing a mask cloth).
- This example is used to illustrate the preparation method of the pineapple extract composition of the present invention.
- Papaya Wash, peel, and dice.
- Oatmeal After crushing, pass through 80 mesh for use.
- oat flour and water are mixed in proportion to 1:50g/mL, and leave standstill at room temperature for 24 hours to obtain oat pretreatment solution.
- the pretreatment solution and the step (1) oat pretreatment solution are mixed according to a ratio of 1:10g/mL, heated at 50°C for 2 hours, and stirred at 100rad/min; cooled to room temperature, filtered to remove impurities, Leave the supernatant (opaque).
- the pineapple extract liquid composition is filled into a mask bag (containing a mask cloth).
- This example is used to illustrate the preparation method of the pineapple extract composition of the present invention.
- Papaya Wash, peel, and dice.
- Oatmeal After crushing, pass through 80 mesh for use.
- the pretreatment solution and the step (1) oat pretreatment solution are mixed in a ratio of 1:10g/mL, heated at 55°C for 1 hour, and stirred at 100rad/min; cooled to room temperature, filtered to remove impurities, Leave the supernatant (opaque).
- the pineapple extract liquid composition is filled into a mask bag (containing a mask cloth).
- This example is used to illustrate the preparation method of the pineapple extract composition of the present invention.
- Papaya Wash, peel, and dice.
- Oatmeal After crushing, pass through 60 mesh for use.
- the pretreatment solution and the step (1) oat pretreatment solution are mixed according to a ratio of 1:20g/mL, heated at 80°C for 3 hours, and stirred at 200rad/min; cooled to room temperature, filtered to remove impurities, Leave the supernatant (opaque).
- the pineapple extract liquid composition is filled into a mask bag (containing a mask cloth). Sterilize at 90°C for 3 hours.
- This example is used to illustrate the preparation method of the pineapple extract composition of the present invention.
- Papaya Wash, peel, and dice.
- Oatmeal After crushing, pass through 90 mesh for use.
- the pretreatment solution and the step (1) oat pretreatment solution are mixed according to a ratio of 1:50g/mL, heated at 85°C for 1 hour, and stirred at 300rad/min; cooled to room temperature, filtered to remove impurities, Leave the supernatant (opaque).
- the homogenizer was homogenized for 50 minutes to obtain the pineapple extract composition.
- the pineapple extract liquid composition is filled into a mask bag (containing a mask cloth).
- This example is used to illustrate the preparation method of the pineapple extract composition of the present invention.
- Papaya Wash, peel, and dice.
- Oatmeal After crushing, pass through 100 mesh for use.
- the pretreatment solution and the step (1) oat pretreatment solution are mixed in a ratio of 1:100g/mL, heated at 90°C for 4 hours, and stirred at 500rad/min; cooled to room temperature, filtered to remove impurities, Leave the supernatant (opaque).
- the homogenizer was homogenized for 60 minutes to obtain the pineapple extract composition.
- the pineapple extract liquid composition is filled into a mask bag (containing a mask cloth).
- This example is used to illustrate the method for preparing an anti-aging cream using the pineapple extract composition of Example 1 of the present invention.
- This example is used to illustrate the method for preparing an anti-aging cream using the pineapple extract composition of Example 7 of the present invention.
- This example is used to prepare anti-aging cream 3 which is compared with Example 8 and Example 9.
- composition of the pineapple extract is replaced with water, and the other ingredients, proportions and operation steps remain unchanged, and the face cream 3 is prepared.
- This test example is used to illustrate the effect of the extract prepared in Example 1 on the activity of glutathione peroxidase (GSH-Px) in fibroblasts.
- the A340 value was recorded every 2 minutes for 10 minutes continuously to obtain 6-point data.
- This test example is used to illustrate the effect of the extract prepared in Example 1 on the content of lipid peroxides in fibroblasts.
- Cell lysate is lysed. Use 0.1 ml of lysate or homogenate per 1 million cells. After lysis, the supernatant was collected by centrifugation at 10,000g-12,000g for 10 minutes for subsequent assays. Sample preparation steps such as lysis should be performed in an ice bath or at 4°C.
- TBA stock solution Preparation of TBA stock solution: Weigh an appropriate amount of TBA and prepare a TBA stock solution with a concentration of 0.37% with TBA preparation solution. For example, 18.5mg TBA is prepared with 5ml TBA preparation solution, and the final concentration is 0.37%.
- the TBA preparation solution needs to be completely dissolved before use. It can be heated to 70°C to promote the dissolution.
- the TBA stock solution is difficult to dissolve and needs to be heated to 70°C and vigorously Vortexed to facilitate dissolution.
- the prepared TBA stock solution should be stored at room temperature away from light, and it is valid for at least 3 months.
- Detection times 1 time 10 times 20 times 50 times TBA Diluent 150 ⁇ l 1500 ⁇ l 3000 ⁇ l 7500 ⁇ l TBA stock solution 50 ⁇ l 500 ⁇ l 1000 ⁇ l 2500 ⁇ l Antioxidants 3 ⁇ l 30 ⁇ l 60 ⁇ l 150 ⁇ l
- Dilution of the standard substance take an appropriate amount of the standard substance and dilute it with distilled water to 1, 2, 5, 10, 20, 50 ⁇ M for subsequent preparation of the standard curve.
- MDA content The molar concentration of MDA can be directly obtained by calculating the sample according to the standard curve. After calculating the MDA content in the sample solution, the MDA content in the original sample can be expressed by the protein content per unit weight, such as ⁇ mol/mg protein. .
- the effect of the pineapple extract composition on the lipid peroxide content in fibroblasts is shown in FIG. 2 .
- the model group was a cell group that was irradiated with UVA only and no sample was added, and the blank group was a cell group that was not irradiated and no sample was added. It was found that UVA irradiation would increase the content of MDA in cells, and there was a statistical difference between the sample group and the model group. Analysis found that the pineapple extract composition can significantly reduce the content of MDA in cells after UVA irradiation, reduce the damage of excessive lipid peroxides to cells, and achieve the effect of anti-aging.
- This test example is used to illustrate the effect of the extraction liquid mask prepared in Experimental Example 2 on the activity of catalase in fibroblasts.
- a Prepare a 250 mM hydrogen peroxide solution.
- c. Prepare color developing working solution. Dissolve the chromogenic substrate on an ice bath, and then use it in appropriate aliquots. Avoid repeated freezing and thawing as much as possible. Other reagents were placed on an ice bath for later use. Take an appropriate amount of peroxidase and dilute it with a chromogenic substrate at a ratio of 1:1000 to prepare a chromogenic working solution. For example, take 5 ⁇ l of peroxidase, add 5 ml of chromogenic substrate, and mix well to obtain 5 ml of working solution for color development.
- Sample preparation lyse cells or tissues with an appropriate lysis buffer to obtain a sample.
- FIG. 3 shows the hydrogen peroxide standard curve in Test Example 3.
- catalase enzyme activity unit 1 enzyme activity unit (1 unit) can catalyze the decomposition of 1 micromolar hydrogen peroxide in 1 minute at 25°C and pH 7.0.
- sample catalase activity [H2O2 consumption in micromoles]X[Dilution factor]/([Reaction minutes]X[Sample volume]X[Protein concentration])
- sample catalase activity is in units/mg protein
- Example volume is X microliters in Table 2, expressed in milliliters is X/1000 milliliters.
- Protein concentration is the protein concentration in the sample when X microliters of sample is taken, and the unit is mg/ml.
- Figure 4 shows the effect of pineapple extract composition on catalase activity in fibroblasts.
- the model group was a cell group that was only irradiated with UVA and no sample was added, and the blank group was a cell group that was not irradiated and no sample was added. It was found that the activity of intracellular catalase decreased after UVA irradiation.
- the pineapple extract composition can significantly increase the enzyme activity of intracellular catalase, and the effect is equivalent to that of the positive control VC. It shows that the pineapple extract composition can increase the degradation of related peroxides in cells, thereby reducing the damage of peroxides to cells and achieving the effect of anti-aging.
- This test example is used to illustrate the effects of the compositions prepared in Examples 1 to 7 on the content of fibroblast collagen (COLI).
- Seeding Seed cells at a seeding density of 4.0 ⁇ 10 4 cells/well into a 24-well plate, and incubate overnight in an incubator (37° C., 5% CO 2 ).
- This test example is used to compare the anti-aging effects of the cream prepared in Example 8, Example 9 and the cream prepared in Example 10 when applied to human body.
- test site has not undergone skin treatment, cosmetic or other tests that may affect the results
- the specified content can be completed according to the requirements of the test plan.
- Test environment temperature: 20 ⁇ 2°C; humidity: (50 ⁇ 10)%, and real-time dynamic monitoring is performed.
- the left face uses the example sample
- the right face uses the comparative sample.
- the skin elasticity (R0/R2) of both cheeks is collected at 0d, 14d, and 28d, respectively, and VISIA CR takes pictures to analyze canthus wrinkles. , fine lines under the eyes, wrinkles on the forehead. Monitoring is required to ensure that subjects correctly distinguish between the test product and the control product on both sides and the continuous use sample.
- Figure 9 shows the effect of the sample and control on the skin R0 in Test Example 5, wherein, *, p ⁇ 0.05, indicating that there is a significant difference compared with the background; n.s., p>0.05, indicating that compared with the background , with no significant difference.
- R0 represents the firmness of the skin, which represents the maximum width of the curve, namely Uf, and the larger it is, the firmer it is. It can be seen from Figure 9 that after the 28-day sample use cycle, the R0 value of the skin of the sample group was significantly higher than the background. The skin R0 value of the control group was not significantly different from the background.
- Figure 10 shows the mean value R2 of the difference between the sample and the control group at each time point and the background in Test Example 5 (comparison between samples), wherein, *, p ⁇ 0.05, indicating that compared with the background, there is a significant Sexual difference; n.s., p>0.05, indicating that there is no significant difference compared with the background.
- R2 represents the total elastic-plasticity of the rebound part/the total elastic-plasticity of the tensile part. Compared with the elastic-plastic properties of the two processes, the closer to 1, the better.
- Figure 10 is the average change of the difference between the sample group (face cream 2) and the control group (face cream 3) and the background. The difference value at the same time point is analyzed for significance.
- FIG. 5 is a graph showing an example of changes in wrinkles (forehead lines) of the volunteer 1 after using the cream 1 prepared in Example 8 in Test Example 5.
- FIG. FIG. 6 is a graph showing an example of changes in wrinkles (canthus) of Volunteer 2 after using Cream 1 prepared in Example 8 in Test Example 5.
- FIG. 7 is a graph showing an example of changes in wrinkles (pockmarks) of Volunteer 1 after using Cream 2 prepared in Example 9 in Test Example 5.
- FIG. 8 is a graph showing an example of changes in wrinkles (pockmarks) of Volunteer 2 after using Cream 2 prepared in Example 9 in Test Example 5.
- FIG. It can be seen from Figure 7 and Figure 8 that the wrinkles were improved at 14d and 28d after using Cream 2.
- the results of the human efficacy test show that the face cream prepared by using the pineapple extract composition of the present invention has a very good anti-wrinkle and firming effect compared to the face cream prepared by adding only water in Example 10 (comparative example), which is embodied in the improvement of skin elasticity. Firmness and improvement of wrinkle levels, etc.
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Abstract
本发明提供了一种具有抗衰老作用的菠萝提取液组合物,其特征在于,所述菠萝提取液组合物由以下成分的原料提取制备得到:菠萝、燕麦、木瓜、罗汉果、葛根粉、植物油和水,还提供了其美容产品、制备方法和应用。本发明人通过试验证明,本发明的菠萝提取液组合物能够增加细胞内相关过氧化物的降解,减少UVA照射后细胞内的MDA含量,增加细胞内相关过氧化物的降解,增加胶原蛋白的含量,从而减少过氧化对细胞的损伤,达到抗衰老的功效。
Description
相关申请的交叉引用
本申请要求2021年04月16日提交的第CN202110412291.5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。
本发明属于化妆品领域,具体涉及一种具有抗衰老作用的菠萝提取液组合物、其制备方法和应用。
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,提供一种具有抗衰老作用的菠萝提取液组合物、其制备方法和应用。
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“GSH-Px”是指:谷胱甘肽过氧化物酶。
术语“TGFβ”是指:转化生长因子-β。
术语“MDA”是指:丙二醛。
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:成纤维细胞毒性实验。
术语“VC”是指:维生素C。
为实现上述目的,本发明的第一方面提供了一种具有抗衰老作用的菠萝提取液组合 物,其特征在于,所述菠萝提取液组合物由以下成分的原料提取制备得到:菠萝、燕麦、木瓜、罗汉果、葛根粉、植物油和水;且所述菠萝提取液组合物通过将燕麦和水制备的燕麦预处理液与含有菠萝、木瓜、罗汉果的混合预处理液混合后再加入葛根粉、植物油制得。
根据本发明第一方面的菠萝提取液组合物,其中,所述植物油选自以下一种或多种:橄榄油、青刺果油、澳洲坚果油、牡丹籽油、椿花油;和/或
所述菠萝提取液组合物中各成分的质量份数为:菠萝0.01~5份,燕麦0.1~10份,木瓜0.01~5份,罗汉果0.01~5份,葛根粉0.5~10份、水100份;
优选地,菠萝0.05~2份,燕麦0.5~5份,木瓜0.1~1份,罗汉果0.1~1份,葛根粉1~8份、水100份;
更优选地,菠萝0.1~1份,燕麦0.5~2份,木瓜0.1~0.5份,罗汉果0.1~0.5份,葛根粉2~5份、水100份。
本发明的第二方面提供了第一方面所述菠萝提取液组合物的制备方法,所述方法包括以下步骤:
(1)将燕麦和水混合静置,得到燕麦预处理液;
(2)菠萝、木瓜、罗汉果分别清洗、去皮、切块,与水混合打浆,室温静置,得到混合预处理液;
(3)将步骤(1)所得燕麦预处理液与步骤(2)所得混合预处理液混合加热搅拌,冷却过滤留上清液;
(4)向步骤(3)中所得上清液中加入葛根粉和植物油,均质得到所述菠萝提取液组合物。
根据本发明第二方面的方法,其中,步骤(1)中,所述燕麦粉碎后过60~100目优选为80目筛;
所述燕麦和水的质量比为1:1~500,优选为1:5~200,更优选为1:20~100,最优选为1:50;和/或
所述静置时间为8~60小时,优选为12~48小时,最优选为24小时。
根据本发明第二方面的方法,其中,步骤(2)中,所述菠萝和水的质量比为1:1~150,优选为1:10~100,更优选为1:20~80,最优选为1:20~50;
所述木瓜和水的质量比为1:1~200,优选为1:10~150,更优选为1:20~100,最优选为1:50~80;
所述罗汉果和水的质量比为1:1~200,优选为1:10~150,更优选为1:20~100,最优选 为1:50~80。
根据本发明第二方面的方法,其中,步骤(3)中,步骤(2)所得混合预处理液与步骤(1)所得燕麦预处理液的质量比为1:1~100,优选为1:1~50,更优选为1:1~20,最优选为1:10。
根据本发明第二方面的方法,其中,步骤(3)中,所述加热温度为20~90℃,优选为25~85℃,更优选为30~80℃,进一步优选为40~60℃;
所述加热时间为0.5~5小时,优选为0.5~4小时,更优选为1~3小时,进一步优选为1~2小时;和/或
所述搅拌速率为50~500r/min,优选为50~300r/min,更优选为50~200r/min,进一步优选为50~150r/min。
根据本发明第二方面的方法,其中,步骤(4)中,所述葛根粉与步骤(3)所得上清液的质量比为1:10~200,优选为1:10~100,更优选为1:20~80,最优选为1:25~50;和/或
所述均质时间为1~60分钟,优选为5~50分钟,更优选为5~30分钟,进一步优选为15~20分钟。
根据本发明第二方面的方法,其中,所述方法还包括以下步骤:
(5)对步骤(4)所得菠萝提取液组合物进行灭菌处理;
优选地,所述灭菌处理方式为加热灭菌;
更优选地,所述加热灭菌温度为60~100℃,优选为70~90℃,最优选为80℃;和/或所述加热灭菌时间为1~5小时,优选为1~3小时,最优选为2小时。
本发明的第三方面提供了一种菠萝提取液组合物在制备抗衰老产品中的应用,所述菠萝提取液组合物包含菠萝、燕麦、木瓜、罗汉果和葛根粉;
优选地,所述菠萝提取液组合物为第一方面所述的菠萝提取液组合物或根据第二方面所述方法制备的菠萝提取液组合物。
本发明的第四方面提供了一种美容产品,所述美容产品包括第一方面所述的菠萝提取液组合物或根据第二方面所述方法制备的菠萝提取液组合物;
优选地,所述美容产品的剂型选自以下一种或多种:膏霜、乳液、水剂、凝胶、油剂、粉剂、泥、气雾剂、贴、膜;
更优选地,所述美容产品为膏霜、面膜、乳液或精华。
根据本发明第四方面的美容产品,其中,当所述美容产品为膏霜时,其制备方法包括以下步骤:
(a)在灭菌后的菠萝提取液组合物中加入水相成分,加热搅拌得水相混合物;
(b)将油相组分混合,加热溶解后得油相混合物;
(c)将步骤(b)制得的油相混合物加入到步骤(a)制备的水相混合物中,均质得所述美容产品。
根据本发明第四方面的美容产品,其中,所述步骤(a)中,所述水相成分包括植物油和抗菌剂;和/或所述步骤(b)中,所述油相成分包括植物油和乳化剂;
优选地,所述植物油选自以下一种或多种:甘油、青刺果油、橄榄油、牡丹籽油、澳洲坚果油、巴巴苏子油、椿花油;
优选地,所述抗菌剂选自以下一种或多种:己二醇、羟苯甲酯、羟苯丙酯、苯氧乙醇;和/或
优选地,所述乳化剂选自以下一种或多种:小麦乳化剂、聚甘油-3二异硬脂酸酯、二聚羟基硬脂酸、耶油基葡糖苷。
根据本发明第四方面的美容产品,其中,所述步骤(a)中,所述灭菌后的菠萝提取液组合物、植物油和抗菌剂的比例为80~95:1~10:0.01~1g/g/g,优选为89~93:2~5:0.05~0.2g/g/g;
所述步骤(b)中,所述植物油和乳化剂的比例为1~10:2~15g/g,优选为2~5:5~10g/g;和/或
所述步骤(c)中,所述油相混合物和所述水相混合物的比例为2~25:0.5~5g/g,优选为5~10:1~2g/g;
优选地,所述步骤(a)、所述步骤(b)和所述步骤(c)中,加热温度为70~120℃,更优选为70~100℃,进一步优选为80~90℃;和/或
优选地,所述步骤(c)中,所述均质的时间为15~45min,更优选为20~40min,进一步优选为25~35min。
本发明的第五方面提供了一种用于抗衰老的方法,所述方法包括:对有需要的受试者给予:
第一方面所述的菠萝提取液组合物;
按照第二方面所述的方法制备的菠萝提取液组合物;或
第四方面所述的美容产品。
本发明菠萝提取液组合物的制备方法包括以下步骤:
(1)将燕麦和水混合制备预处理液。室温静置24小时。
(2)将菠萝、木瓜、罗汉果与水混合打浆,加入(1)中,加热搅拌,过滤。
(3)将葛根粉加入(2)提取液中。再加入植物油均质得到所述菠萝提取液组合物。
(4)100℃ 2h灭菌,无菌条件下灌装。
步骤(1)中,所述燕麦和水的比例为1:1~500g/mL,优选为1:5~300g/mL,更优选为1:20~200g/mL,最优选为1:100g/mL;
步骤(2)中,所述菠萝和水的比例为1:1~150g/mL,优选为1:10~100g/mL,更优选为1:20~80g/mL,最优选为1:50g/mL;
所述木瓜和水的比例为1:1~200g/mL,优选为1:10~150g/mL,更优选为1:20~100g/mL,最优选为1:80g/mL;
所述罗汉果和水的比例为1:1~200g/mL,优选为1:10~150g/mL,更优选为1:50~100g/mL,最优选为1:80g/mL;
所述加热温度为1~150℃,优选为10~100℃,更优选为20~100℃,最优选为40℃;
所述加热时间为1~10小时,优选为1-8小时,更优选为1~5小时,最优选为4小时
所述搅拌速率为10~600r/min,优选为10~500r/min,更优选为10~300r/min,最优选为150r/min。
步骤(3)中,所述葛根粉和提取液的比例为1:1~100g/mL,优选为1:5~80g/mL,更优选为1:10~70g/mL,最优选为1:50g/mL;
植物油的加入量为1~30%,优选为1~20%;更优选为1~10%,最优选为3%;
所述均质速度为1000~8000r/min,优选为1000~7000r/min,更优选为2000~6000r/min;最优为5000r/min;
均质时间为1~60分钟,优选为1~50分钟,更优选为10~40分钟,最优选为15分钟。
本发明的菠萝提取液组合物可以具有但不限于以下有益效果:
本发明提供了一种菠萝提取液组合物在制备具有抗衰老作用的产品中的应用,本发明人通过试验证明,本发明的菠萝提取液组合物能够增加细胞内相关过氧化物的降解,减少UVA照射后细胞内的MDA含量,增加细胞内相关过氧化物的降解,从而减少过氧化对细胞的损伤,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示出了试验例1中对成纤维细胞内谷胱甘肽过氧化物酶(GSH-Px)的影响。
图2示出了试验例2中成纤维细胞内脂质过氧化物含量的影响。
图3示出了试验例3中过氧化氢标准曲线。
图4示出了试验例3中过氧化氢酶酶活力的影响。
图5示出了试验例5中使用实施例8制得的面霜1后的志愿者1的皱纹(抬头纹)变化个例图。
图6示出了试验例5中使用实施例8制得的面霜1后的志愿者2的皱纹(眼角纹)变化个例图。
图7示出了试验例5中使用实施例9制得的面霜2后的志愿者1的皱纹(眼袋纹)变化个例图。
图8示出了试验例5中使用实施例9制得的面霜2后的志愿者2的皱纹(眼袋纹)变化个例图。
图9示出了试验例5中样品和对照对皮肤R0的影响,其中,*,p<0.05,表示与本底相比,具有显著性差异;n.s.,p>0.05,表示与本底相比,无显著性差异。
图10示出了试验例5中样品和对照组各时间点下与本底的差值平均值R2(样品之间比较),其中,*,p<0.05,表示与本底相比,具有显著性差异;n.s.,p>0.05,表示与本底相比,无显著性差异。
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、木瓜、罗汉果,购自广东省湛江市红星农场;
燕麦(炒制熟)购自张家口建军燕麦食品有限公司;
葛根(干燥粉)购自中国北京同仁堂(集团)有限责任公司。
试剂:
试剂盒,购自北京索莱宝科技有限公司;
VC,购自碧云天生物技术有限公司;
成纤维细胞,赛百慷(上海)生物技术股份有限公司。
仪器:
酶联免疫检测仪,购自Thermo Fisher Scientific OY有限公司、型号1510-00662C。
均质机,购自IKA有限公司,型号T25。
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。
皮肤弹性测试仪器,购自北京金宏帆商贸有限公司,型号MPA580多探头。
面部图像分析仪,购自北京金宏帆商贸有限公司,型号VISIA CR。
实施例1
本实施例用于说明本发明菠萝提取液组合物的制备方法。
1、原料处理
菠萝:清洗、去皮、切块。
木瓜:清洗、去皮、切块。
罗汉果:清洗、去皮、切块。
燕麦:粉碎后过80目备用。
2、菠萝提取液组合物制备
(1)燕麦粉与水按照比例为1:50g/mL混合,室温静置24小时,得到燕麦预处理液。
(2)制备混合预处理液,其中,菠萝、木瓜、罗汉果与水添加比例如表1所示,将菠萝、木瓜、罗汉果与水混合打浆,静置2小时,即为混合预处理液。
表1混合预处理液的添加比例
原料 | 质量份数 |
水 | 100 |
菠萝 | 5 |
木瓜 | 2 |
罗汉果 | 2 |
(3)步骤(2)中预处理液与步骤(1)燕麦预处理液按照比例为1:10g/mL混合,40℃加热2小时,50rad/min进行搅拌;冷却至室温,过滤去除杂质,留上清液(不透明)。
(4)加入与上清液比例为1:25g/mL的葛根粉,质量比为2%橄榄油,均质机均质15分钟,得到所述菠萝提取液组合物。
3、灭菌处理
菠萝提取液组合物灌装入面膜袋(含面膜布)。
80℃灭菌2小时。
实施例2
本实施例用于说明本发明菠萝提取液组合物的制备方法。
1、原料处理(食品级)
菠萝:清洗、去皮、切块。
木瓜:清洗、去皮、切块。
罗汉果:清洗、去皮、切块。
燕麦:粉碎后过80目备用。
2、菠萝提取液组合物制备
(1)燕麦粉与水按照比例为1:100g/mL混合,室温静置24小时,得到燕麦预处理液。
(2)制备混合预处理液,其中,菠萝、木瓜、罗汉果与水添加比例如表2所示,将菠萝、木瓜、罗汉果与水混合打浆,静置2小时,即为混合预处理液。
表2预处理液的添加比例
原料 | 质量份数 |
水 | 100 |
菠萝 | 2 |
木瓜 | 1.25 |
罗汉果 | 1.25 |
(3)步骤(2)中预处理液与步骤(1)燕麦预处理液按照比例为1:10g/mL混合,60℃加热1小时,150rad/min进行搅拌;冷却至室温,过滤去除杂质,留上清液(不透明)。
(4)加入与上清液比例为1:50g/mL的葛根粉,3%橄榄油,均质机均质20分钟。
3、灭菌处理
菠萝提取液组合物灌装入面膜袋(含面膜布)。
80℃灭菌2小时。
实施例3
本实施例用于说明本发明菠萝提取液组合物的制备方法。
1、原料处理
菠萝:清洗、去皮、切块。
木瓜:清洗、去皮、切块。
罗汉果:清洗、去皮、切块。
燕麦:粉碎后过80目备用。
2、菠萝提取液组合物制备
(1)燕麦粉与水按照比例为1:50g/mL混合,室温静置24小时,得到燕麦预处理 液。
(2)制备混合预处理液,其中,菠萝、木瓜、罗汉果与水添加比例如表3所示,将菠萝、木瓜、罗汉果与水混合打浆,静置2小时,即为混合预处理液。
表3混合预处理液添加比例
原料 | 质量份数 |
水 | 100 |
菠萝 | 4 |
木瓜 | 1.25 |
罗汉果 | 1.25 |
(3)步骤(2)中预处理液与步骤(1)燕麦预处理液按照比例为1:10g/mL混合,50℃加热2小时,100rad/min进行搅拌;冷却至室温,过滤去除杂质,留上清液(不透明)。
(4)加入与上清液比例为1:30g/mL的葛根粉,质量比为4%青刺果油,
均质机均质20分钟,得到所述菠萝提取液组合物。
3、灭菌处理
菠萝提取液组合物灌装入面膜袋(含面膜布)。
80℃灭菌2小时。
实施例4
本实施例用于说明本发明菠萝提取液组合物的制备方法。
1、原料处理
菠萝:清洗、去皮、切块。
木瓜:清洗、去皮、切块。
罗汉果:清洗、去皮、切块。
燕麦:粉碎后过80目备用。
2、菠萝提取液组合物制备
(1)燕麦粉与水按照比例为1:50g/mL混合,室温静置24小时,得到燕麦预处理液。
(2)制备混合预处理液,其中,菠萝、木瓜、罗汉果与水添加比例如表4所示,将菠萝、木瓜、罗汉果与水混合打浆,静置2小时,即为混合预处理液。
表4混合预处理液添加比例
原料 | 质量份数 |
水 | 100 |
菠萝 | 3 |
木瓜 | 2 |
罗汉果 | 2 |
(3)步骤(2)中预处理液与步骤(1)燕麦预处理液按照比例为1:10g/mL混合,55℃加热1小时,100rad/min进行搅拌;冷却至室温,过滤去除杂质,留上清液(不透明)。
(4)加入与上清液比例为1:40g/mL的葛根粉,质量比为2%澳洲坚果油,
均质机均质15分钟,得到所述菠萝提取液组合物。
3、灭菌处理
菠萝提取液组合物灌装入面膜袋(含面膜布)。
80℃灭菌2小时。
实施例5
本实施例用于说明本发明菠萝提取液组合物的制备方法。
1、原料处理
菠萝:清洗、去皮、切块。
木瓜:清洗、去皮、切块。
罗汉果:清洗、去皮、切块。
燕麦:粉碎后过60目备用。
2、菠萝提取液组合物制备
(1)燕麦粉与水按照比例为1:20g/mL混合,室温静置48小时,得到燕麦预处理液。
(2)制备混合预处理液,其中,菠萝、木瓜、罗汉果与水添加比例如表5所示,将菠萝、木瓜、罗汉果与水混合打浆,静置3小时,即为混合预处理液。
表5混合预处理液添加比例
原料 | 质量份数 |
水 | 100 |
菠萝 | 5 |
木瓜 | 5 |
罗汉果 | 5 |
(3)步骤(2)中预处理液与步骤(1)燕麦预处理液按照比例为1:20g/mL混合,80℃加热3小时,200rad/min进行搅拌;冷却至室温,过滤去除杂质,留上清液(不透明)。
(4)加入与上清液比例为1:80g/mL的葛根粉,质量比为2%牡丹籽油,
均质机均质30分钟,得到所述菠萝提取液组合物。
3、灭菌处理
菠萝提取液组合物灌装入面膜袋(含面膜布)。90℃灭菌3小时。
实施例6
本实施例用于说明本发明菠萝提取液组合物的制备方法。
1、原料处理
菠萝:清洗、去皮、切块。
木瓜:清洗、去皮、切块。
罗汉果:清洗、去皮、切块。
燕麦:粉碎后过90目备用。
2、菠萝提取液组合物制备
(1)燕麦粉与水按照比例为1:200g/mL混合,室温静置60小时,得到燕麦预处理液。
(2)制备混合预处理液,其中,菠萝、木瓜、罗汉果与水添加比例如表6所示,将菠萝、木瓜、罗汉果与水混合打浆,静置5小时,即为混合预处理液。
表6混合预处理液添加比例
原料 | 质量份数 |
水 | 100 |
菠萝 | 10 |
木瓜 | 10 |
罗汉果 | 10 |
(3)步骤(2)中预处理液与步骤(1)燕麦预处理液按照比例为1:50g/mL混合,85℃加热1小时,300rad/min进行搅拌;冷却至室温,过滤去除杂质,留上清液(不透明)。
(4)加入与上清液比例为1:100g/mL的葛根粉,质量比为4%橄榄油,
均质机均质50分钟,得到所述菠萝提取液组合物。
3、灭菌处理
菠萝提取液组合物灌装入面膜袋(含面膜布)。
100℃灭菌1小时。
实施例7
本实施例用于说明本发明菠萝提取液组合物的制备方法。
1、原料处理
菠萝:清洗、去皮、切块。
木瓜:清洗、去皮、切块。
罗汉果:清洗、去皮、切块。
燕麦:粉碎后过100目备用。
2、菠萝提取液组合物制备
(1)燕麦粉与水按比例为1:500g/mL混合,室温静置50小时,得到燕麦预处理液。
(2)制备混合预处理液,其中,菠萝、木瓜、罗汉果与水添加比例如表7所示,将菠萝、木瓜、罗汉果与水混合打浆,静置5小时,即为混合预处理液。
表7混合预处理液添加比例
原料 | 质量份数 |
水 | 100 |
菠萝 | 50 |
木瓜 | 30 |
罗汉果 | 25 |
(3)步骤(2)中预处理液与步骤(1)燕麦预处理液按照比例为1:100g/mL混合,90℃加热4小时,500rad/min进行搅拌;冷却至室温,过滤去除杂质,留上清液(不透明)。
(4)加入与上清液比例为1:200g/mL的葛根粉,质量比为1.5%椿花油,
均质机均质60分钟,得到所述菠萝提取液组合物。
3、灭菌处理
菠萝提取液组合物灌装入面膜袋(含面膜布)。
85℃灭菌2小时。
实施例8
本实施例用于说明利用本发明实施例1菠萝提取液组合物制备抗衰老面霜的方法。
1、制备抗衰老面霜,其中面霜的成分及比例如表8所示。
表8面霜成分及添加比例
2、将表8中水相成分混合,在85℃加热30min,250rad/min进行搅拌,得到混合物1。
3、将表8中油相组分混合,在85℃下加热至溶解,得到混合物2。
4、在85℃环境下将混合物2加入混合物1中,均质30min,得到抗衰老面霜1。
实施例9
本实施例用于说明利用本发明实施例7菠萝提取液组合物制备抗衰老面霜的方法。
1、制备抗衰老面霜,其中面霜的成分及比例如表9所示。
表9面霜成分及添加比例
2、将表9中水相成分混合,在85℃加热30min,250rad/min进行搅拌,得到混合物1。
3、将表9中油相组分混合,在85℃下加热至溶解,得到混合物2。
4、在85℃环境下将混合物2加入混合物1中,均质30min,得到抗衰老面霜2。
实施例10
本实施例用于制备和实施例8、实施例9做对比的抗衰老面霜3。
区别在于把菠萝提取液组合物替换成水,其他成分及比例和操作步骤不变,制得面霜3。
试验例1
本试验例用于说明实施例1制备提取液对成纤维细胞内谷胱甘肽过氧化物酶(GSH-Px)活力的影响。
1)细胞裂解液上清制备
取对数生长期状态良好的成纤维细胞计数并接种于6孔培养板中,控制每孔细胞数5x10
5个细胞。在37℃,5%CO
2环境下培养过夜,弃培养基,加入少量的PBS(pH=7.4)以刚好覆盖细胞为宜,随后以UVA刺激细胞,UV辐射量为7J/cm
2,空白组不照射。吸弃PBS,加入不同浓度(使成纤维细胞存活率为50%、80%、100%的样品浓度)样品作用细胞24小时(模型组以及空白对照组加入无血清DMEM培养液)。取出,置于冰上,PBS洗涤2次;以细胞刮刀刮下细胞,收集至离心管中,5000r/min离心5min弃上清得到细胞沉淀,后加入200μL裂解液裂解细胞,12000g 4℃下离心5min,取上清得到细胞 裂解上清液。
2)试剂盒的准备工作:
a.配制30mM NADPH。在本试剂盒提供的15mg NADPH中加入600微升超纯水,溶解,混匀。
b.84mM GSH溶液的配制。在14mg GSH中加入550微升超纯水,溶解并混匀。溶液。
c.GPx检测工作液的配制。
表10 GPx检测工作液的配制
可测定样品数(含对照) | 1个样品 |
30mM NADPH | 5μL |
84mM GSH | 5μL |
谷胱甘肽还原酶 | 1.6μL |
GPx检测工作液体积 | 11.6μL |
d.15mM过氧化物试剂溶液的配制。取21.5微升过氧化物试剂(t-Bu-OOH)加入10毫升超纯水,混匀,即配制成15mM过氧化物试剂溶液。
3)样品测定
参考下表4,使用96孔板,依次加入检测缓冲液、待测样品和GPx检测工作液,混匀。加入4微升15mM过氧化物试剂溶液后,反应开始。需适当混匀。
表11各溶液加入量
空白对照(blank) | 样品本底对照 | 样品 | |
谷胱甘肽还原酶 | 185μL | 179-187μL | 175-183μL |
待测样品 | — | 2-10μL | 2-10μL |
GPx检测工作液 | 11μL | 11μL | 11μL |
15mM过氧化物试剂溶液 | 4μL | — | 4μL |
总体积 | 200μL | 200μL | 200μL |
使用荧光酶标仪,每隔2分钟记录A 340值,连续记录10分钟,获得6个点的数据。
结果:菠萝提取液组合物对成纤维细胞内谷胱甘肽过氧化物酶酶活力的影响如图1所示。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,经试验发现UVA照射使细胞的谷胱甘肽过氧化物酶活力下降,样品组与模型组做统计学差异后发现,本菠萝提取液组合物能够显著提高成纤维细胞的谷胱甘肽过氧化物酶活力,甚至高于阳性对照VC,说明发明提取液能够增加细胞内相关过氧化物的降解,从而减少过氧化对细胞的损伤,达到抗衰老的功效。
试验例2
本试验例用于说明实施例1制备提取液对成纤维细胞内脂质过氧化物含量的影响。
1)样品的准备:
a.细胞裂解液进行裂解。每100万细胞使用0.1ml裂解液或匀浆液。裂解后,10,000g-12,000g离心10分钟取上清用于后续测定。裂解等样品制备步骤宜在冰浴或4℃进行操作。
2)试剂盒的准备工作:
a.TBA储存液的配制:称取适量TBA,用TBA配制液配制成浓度为0.37%的TBA储存液。例如18.5mg TBA用5ml TBA配制液配制,最终浓度即为0.37%。TBA配制液需完全溶解后再使用,可以加热到70℃以促进溶解。TBA储存液较难溶解,需加热到70℃,并通过剧烈Vortex以促进溶解。配制好的TBA储存液室温避光保存,至少3个月内有效。
b.MDA检测工作液的配制:根据待测定的样品数(含对照),参考下表12在临检测前新鲜配制适量的MDA检测工作液。
表12 MDA检测工作液的配制
检测次数 | 1次 | 10次 | 20次 | 50次 |
TBA稀释液 | 150μl | 1500μl | 3000μl | 7500μl |
TBA储存液 | 50μl | 500μl | 1000μl | 2500μl |
抗氧化剂 | 3μl | 30μl | 60μl | 150μl |
c.标准品的稀释:取适量标准品用蒸馏水稀释至1、2、5、10、20、50μM,用于后续制作标准曲线。
3)样品测定:
a.在离心管或其它适当容器内加入0.1ml匀浆液、裂解液或PBS等适当溶液作为空白对照,加入0.1ml上述不同浓度标准品用于制作标准曲线,加入0.1ml样品用于测定;随后加入0.2ml MDA检测工作液。参考下表设置检测反应体系:
表13反应体系的设置
空白对照 | 标准品 | 样品 | |
匀浆液、裂解液或PBS | 0.1ml | - | - |
标准品 | - | 0.1ml | - |
待测样品 | - | - | 0.1ml |
MDA检测工作液 | 0.2ml | 0.2ml | 0.2ml |
b.混匀后,100℃沸水浴加热15分钟。加热时务必注意避免液体暴沸溅出。使用沸水浴,则需使用可把盖子锁死的离心管或螺旋盖离心管,或用Parafilm封住离心管口,用针头刺一小孔。
c.水浴冷却至室温,1000g室温离心10分钟。取200微升上清加入到96孔板中,随后用酶标仪在532nm测定吸光度。如果不方便测定532nm的吸光度,也可以测定530-540nm之间的吸光度。
d.MDA含量的计算:样品可以直接根据标准曲线计算获得MDA的摩尔浓度,计算出样品溶液中的MDA含量后,可以通过单位重量的蛋白含量表示最初样品中的MDA含量,例如μmol/mg蛋白。
结果:菠萝提取液组合物对成纤维细胞内的脂质过氧化物含量的影响如图2所示。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,经试验发现UVA照射会增加细胞内MDA的含量,样品组与模型组做统计学差异分析发现,本菠萝提取液组合物能极其显著的减少UVA照射后细胞内的MDA含量,减少过量的脂质过氧化物对细胞的损伤,达到抗衰老的功效。
试验例3
本试验例用于说明实验例2制备提取液面膜对成纤维细胞内过氧化氢酶活力的影响。
1)准备工作:
a.配制250mM过氧化氢溶液。试剂盒提供的过氧化氢浓度约为1M。由于过氧化氢不是非常稳定,使用前需自行测定过氧化氢的实际浓度。把浓度约为1M的过氧化氢用试剂盒提供的过氧化氢酶检测缓冲液稀释100倍,使过氧化氢的浓度约为10mM。测定A240。过氧化氢浓度(mM)=22.94×A240。从而计算出本试剂盒提供的过氧化氢的实际浓度。然后再根据实际的过氧化氢浓度配制250mM过氧化氢溶液。
b.配制5mM过氧化氢溶液。根据测定得到的实际过氧化氢浓度配制5mM过氧化氢溶液。
c.配制显色工作液。在冰浴上溶解显色底物,适当分装后再使用,尽量避免反复冻融。其它试剂放置在冰浴上备用。取适当量的过氧化物酶,按照1:1000的比例用显色底物稀释,配制成显色工作液。例如取5μl过氧化物酶,加入5ml显色底物,混匀即得到5ml显色工作液。
2)样品准备:用适当的裂解液裂解细胞或组织,得到样品。
3)标准曲线测定:
a.取0、12.5、25、50、75微升配制好的5mM过氧化氢溶液至1.5ml塑料离心管中,分别加入过氧化氢酶检测缓冲液至最后体积为100微升,混匀。
b.各取4微升,加入到96孔板中的一个孔内。加入200μl显色工作液。25℃至少孵育15分钟后测定A520,但孵育时间不宜超过45分钟。
4)样品测定
表14各溶液的添加量
空白对照(blank) | 样品(sample) | |
样品体积 | 0μl | 4μl |
过氧化氢酶检测缓冲液 | 40μl | 36μl |
250mM过氧化氢溶液 | 10μl | 10μl |
a.参考表14,取4微升样品至1.5ml塑料离心管中,加入过氧化氢酶检测缓冲液至体积为40微升,混匀。再加入10微升250mM过氧化氢溶液,用移液器迅速混匀。参考表1,25℃反应5分钟。
b.加入450微升过氧化氢酶反应终止液,颠倒混匀或Vortex混匀以终止反应。需在终止反应后15分钟内完成下面的步骤c和步骤d。
c.在一洁净的塑料离心管内加入40微升过氧化氢酶检测缓冲液,再加入10微升已终止并混匀的上述反应体系,混匀。
d.从上一步骤的50微升体系中取10微升加入到96孔板中的一个孔内。加入200微升显色工作液。
e.25℃至少孵育15分钟后测定A520,但孵育时间不宜超过45分钟。
5)样品中过氧化氢酶酶活力的计算:
a.计算出标准曲线。A520=k[过氧化氢微摩尔数]+b,由标准曲线计算出k和b的值。图3示出了试验例3中过氧化氢标准曲线。
b.计算出样品中残余的过氧化氢微摩尔数。
残余过氧化氢微摩尔数=(A520-b)/k
c.过氧化氢酶酶活力单位的定义:1个酶活力单位(1unit)在25℃,pH7.0的条件下,在1分钟内可以催化分解1微摩尔过氧化氢。
d.对于细胞或组织样品的过氧化氢酶活力计算:
[样品过氧化氢酶酶活力]=[消耗过氧化氢微摩尔数]X[稀释倍数]/([反应分钟数]X[样品体积]X[蛋白浓度])
[样品过氧化氢酶酶活力]的单位为units/mg蛋白
[消耗过氧化氢微摩尔数]=[空白对照残余过氧化氢微摩尔数]-[样品残余过氧化氢微摩尔数]
[稀释倍数]=250
[反应分钟数]即为实际的反应分钟数
[样品体积]为表2中的X微升,以毫升来表示即为X/1000毫升。
[蛋白浓度]为取X微升样品时,样品中的蛋白浓度,单位为mg/ml。
结果:菠萝提取液组合物对成纤维细胞内过氧化氢酶酶活力的影响如图4所示。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,经试验发现UVA照射后,细胞内过氧化氢酶酶活力会降低,样品组与模型组做统计学差异分析后可知,本菠萝提取液组合物能够极其显著的增加细胞内过氧化氢酶的酶活力,效果与阳性对照VC相当。说明本面菠萝提取液组合物能够增加细胞内相关过氧化物的降解,从而减少过氧化对细胞的损伤,达到抗衰老的功效。
试验例4
本试验例用于说明实施例1~7制备的组合物液对成纤维细胞胶原蛋白(COLI)含量的影响。
测试方案如表15所示
表15测试方案
具体操作步骤如下:
(1)接种:按4.0×10
4个/孔的接种密度接种细胞至24孔板,培养箱(37℃、5%CO
2)中孵育过夜。
(2)给药:根据表15测试方案,待24孔板中细胞铺板率达到40%~60%时,进行分组给药,每孔加样1mL,每组设3个复孔。给药完成后将24孔板放置在培养箱(37℃、5%CO
2)中培养24h。
(3)收样:孵育培养结束后,吸弃旧液,1mL/孔PBS清洗三次,每孔加1mL 4%的多聚甲醛室温固定15min。
(4)封闭:滴加山羊血清200μL/孔,室温封闭60min。
(5)孵育一抗:弃掉山羊血清封闭液,滴加适当比例稀释后的一抗(200μL/孔)(山羊血清稀释),4℃孵育过夜。
(6)孵育二抗:用PBS清洗3次/5min,滴加适当比例稀释后的荧光二抗(200μL/孔),室温避光孵育1h。
(7)核染:用PBS清洗3次/5min,滴加Hochest33342(200μL/孔),室温孵育5min。
(8)封片:用针头挑出爬片,在载玻片上滴加一滴抗淬灭剂并将爬片反放与载玻片上。
(9)观察拍照:24h内荧光显微镜拍照。
结果:实施例制备的面膜液对成纤维细胞胶原蛋白(COLI)积分光密度(IOD)值 如表16所示。从表中可以看出,UVA辐射后成纤维细胞中COLI含量显著下降,经实施例制得得样品处理后COLI含量显著增加,说明本发明制备的组合物具有较好的抗衰老功效。
表16胶原蛋白(COLI)积分光密度(IOD)
样品名称 | 相对IOD/细胞数平均值 | SD | P-value |
空白对照BC | 1.00 | 0.07 | / |
阴性对照NC | 0.77 | 0.03 | 0.007## |
阳性对照PC(TGFβ) | 1.27 | 0.04 | 0.000** |
实施例1-2.5% | 1.74 | 0.25 | 0.000** |
实施例2-10% | 1.95 | 0.24 | 0.001** |
实施例3-2.5% | 3.36 | 0.52 | 0.001** |
实施例4-10% | 1.99 | 0.30 | 0.001** |
实施例5-10% | 1.65 | 0.12 | 0.000** |
实施例6-10% | 1.35 | 0.06 | 0.000** |
实施例7-10% | 1.28 | 0.12 | 0.002** |
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**。
试验例5(对比例)
本试验例用于对比实施例8、实施例9制备的面霜和实施例10制备的面霜应用于人体的抗衰老功效。
1、基本原则
根据赫尔辛基宣言,志愿者的选择遵循人体测试的医学和伦理学标准,所有志愿者的测试都必须出于志愿者本人自愿,并在测试前签署知情同意书。志愿者在签署知情同意书之前,测试人员需要告知志愿者本次测试的目的、可能的利益、潜在的风险和问题以及相关的权利和义务。
2、志愿者筛选
符合下列所有条件的志愿者将被入选。
(1)年龄45~65岁的志愿者,干性皮肤、皮肤松弛、皱纹等级3级及以上,左右两侧颧骨下方F4平均值高于6,R2平均值低于0.65,男女均可,其中妊娠或哺乳期妇女,或六个月内预备怀孕妇女除外;
(2)受试部位没有接受过皮肤治疗、美容以及其他可能影响结果的测试;
(3)近一个月内未曾使用激素类药物及免疫抑制剂者;
(4)现在或最近三个月受试部位未参加其他临床试验者。
(5)自愿参加并签署知情同意书。
(6)能按测试方案要求完成规定内容。
3、实验设计
(1)测试环境:温度:20±2℃;湿度:(50±10)%,并且进行实时动态监测。
(2)有效人数:45~65岁60人,30人使用面霜1和面霜3,30人使用面霜2和面霜3.
(3)实验仪器:MPA580(德国CK公司)多探头(Cutometer皮肤弹性测试仪器参数设置为:负压45kPa(450mbar),10个循环周期,负压加载和清除时间均为2s),VISIA CR
(4)测试区域:面部
(5)测试时间点及指标:
连续4周使用,早晚各一次,左脸使用实施例样品,右脸使用对比例样品,分别在0d,14d,28d时采集两侧脸颊的皮肤弹性(R0/R2),VISIA CR拍照分析眼角皱纹、眼下细纹、抬头纹。需采用能够确保受试者正确区分两侧试验产品和对照产品和连续使用样品的监控措施。
(6)数据分析:
统计学方法:正态数据用均值、标准差进行描述,比较用t检验;非正态数据用中位数和四分位数间距进行描述,比较用非参数检验。以p<0.05表示差异有统计学意义。
结果:
图9示出了试验例5中样品和对照对皮肤R0的影响,其中,*,p<0.05,表示与本底相比,具有显著性差异;n.s.,p>0.05,表示与本底相比,无显著性差异。R0表示皮肤紧实度,代表曲线的最大宽度,即Uf,越大表示越紧实。由图9可知,经过28天的样品使用周期,样品组皮肤R0值显著性高于本底。对照组皮肤R0值与本底相比无显著性差异。
图10示出了试验例5中样品和对照组各时间点下与本底的差值平均值R2(样品之间比较),其中,*,p<0.05,表示与本底相比,具有显著性差异;n.s.,p>0.05,表示与本底相比,无显著性差异。R2表示回弹部分的弹塑性总量/拉伸部分的弹塑性总量,两个过程的弹塑性性能相比,越接近1越好。图10为样品组(面霜2)和对照组(面霜3)与本底的差值平均值变化,相同时间点下差值进行显著性分析,如与对照组相比具有显著性差异,表示样品具有显著改变R2的作用。由图可知,经过28天的样品使用周期,样品组的各时间点R2与本底的差值>0,表示样品使用后各个时间点R2值升高;对照组的各时间点R2与本底的差值<0,表示对照使用后各个时间点R2值降低。通过对相同时 间点下差值进行统计学分析发现,14D时样品与对照两组差值无显著性差异,28D时样品组差值显著高于对照组差值(P<0.05),表示样品与对照相比,具有提升R2值的作用。
图5示出了试验例5中使用实施例8制得的面霜1后的志愿者1的皱纹(抬头纹)变化个例图。图6示出了试验例5中使用实施例8制得的面霜1后的志愿者2的皱纹(眼角纹)变化个例图。从图5和图6中可以看出,使用面霜2后14d,28d皱纹都得到了改善。
图7示出了试验例5中使用实施例9制得的面霜2后的志愿者1的皱纹(眼袋纹)变化个例图。图8示出了试验例5中使用实施例9制得的面霜2后的志愿者2的皱纹(眼袋纹)变化个例图。从图7和图8可以看出,使用面霜2后14d,28d皱纹都得到了改善。
人体功效试验结果说明本发明的使用菠萝提取液组合物制备的面霜相较于实施例10(对比例)仅添加水制备的面霜,具有很好的抗皱紧致功效,具体表现在改善皮肤弹性提升紧实度和改善皱纹等级等方面。
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)
- 一种具有抗衰老作用的菠萝提取液组合物,其特征在于,所述菠萝提取液组合物由以下成分的原料提取制备得到:菠萝、燕麦、木瓜、罗汉果、葛根粉、植物油和水;且所述菠萝提取液组合物通过将燕麦和水制备的燕麦预处理液与含有菠萝、木瓜、罗汉果的混合预处理液混合后再加入葛根粉、植物油制得。
- 根据权利要求1所述的菠萝提取液组合物,其特征在于,所述植物油选自以下一种或多种:橄榄油、青刺果油、澳洲坚果油、牡丹籽油、椿花油;和/或所述菠萝提取液组合物中各成分的质量份数为:菠萝0.01~5份,燕麦0.1~10份,木瓜0.01~5份,罗汉果0.01~5份,葛根粉0.5~10份、水100份;优选地,菠萝0.05~2份,燕麦0.5~5份,木瓜0.1~1份,罗汉果0.1~1份,葛根粉1~8份、水100份;更优选地,菠萝0.1~1份,燕麦0.5~2份,木瓜0.1~0.5份,罗汉果0.1~0.5份,葛根粉2~5份、水100份。
- 根据权利要求1或2所述的菠萝提取液组合物的制备方法,其特征在于,所述菠萝提取液组合物通过以下方法制备:(1)将燕麦和水混合静置,得到燕麦预处理液;(2)菠萝、木瓜、罗汉果分别清洗、去皮、切块,与水按比例混合后打浆静置,得到混合预处理液;优选地,所述静置时间为1~5小时,更优选为1~3小时,最优选为2小时;(3)将步骤(1)所得燕麦预处理液与步骤(2)所得混合预处理液混合加热搅拌,冷却过滤留上清液;(4)向步骤(3)中所得上清液中加入葛根粉和植物油,均质得到所述菠萝提取液组合物。
- 根据权利要求3所述的方法,其特征在于,步骤(1)中,所述燕麦粉碎后过60~100目优选为80目筛;所述燕麦和水的质量比为1:1~500,优选为1:5~200,更优选为1:20~100,最优选为1:50;和/或所述静置时间为8~60小时,优选为12~48小时,最优选为24小时。
- 根据权利要求3或4所述的方法,其特征在于,步骤(2)中,所述菠萝和水的质量比为1:1~150,优选为1:10~100,更优选为1:20~80,最优选为1:20~50;所述木瓜和水的质量比为1:1~200,优选为1:10~150,更优选为1:20~100,最优选为1:50~80;所述罗汉果和水的质量比为1:1~200,优选为1:10~150,更优选为1:20~100,最优选为1:50~80。
- 根据权利要求3至5中任一项所述的方法,其特征在于,步骤(3)中,步骤(2)所得混合预处理液与步骤(1)所得燕麦预处理液的质量比为1:1~100,优选为1:1~50,更优选为1:1~20,最优选为1:10。
- 根据权利要求3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20~90℃,优选为25~85℃,更优选为30~80℃,进一步优选为40~60℃;所述加热时间为0.5~5小时,优选为0.5~4小时,更优选为1~3小时,进一步优选为1~2小时;和/或所述搅拌速率为50~500r/min,优选为50~300r/min,更优选为50~200r/min,进一步优选为50~150r/min。
- 根据权利要求3至7中任一项所述的方法,其特征在于,步骤(4)中,所述葛根粉与步骤(3)所得上清液的质量比为1:10~200,优选为1:10~100,更优选为1:20~80,最优选为1:25~50;和/或所述均质时间为1~60分钟,优选为5~50分钟,更优选为5~30分钟,进一步优选为15~20分钟。
- 根据权利要求3至8中任一项所述的方法,其特征在于,所述方法还包括以下步骤:(5)对步骤(4)所得菠萝提取液组合物进行灭菌处理;优选地,所述灭菌处理方式为加热灭菌;更优选地,所述加热灭菌温度为60~100℃,优选为70~90℃,最优选为80℃;和/或所述加热灭菌时间为1~5小时,优选为1~3小时,最优选为2小时。
- 一种菠萝提取液组合物在制备抗衰老产品中的应用,其特征在于,所述菠萝提取液组合物包含菠萝、燕麦、木瓜、罗汉果和葛根粉;优选地,所述菠萝提取液组合物为权利要求1或2所述的菠萝提取液组合物或根据权利要求3至9中任一项所述方法制备的菠萝提取液组合物。
- 一种美容产品,其特征在于,所述美容产品包括权利要求1或2所述的菠萝提取液组合物或根据权利要求3至9中任一项所述方法制备的菠萝提取液组合物;优选地,所述美容产品的剂型选自以下一种或多种:膏霜、乳液、水剂、凝胶、油剂、粉剂、泥、气雾剂、贴、膜;更优选地,所述美容产品为膏霜、面膜、乳液或精华。
- 根据权利要求11所述的美容产品,其特征在于:当所述美容产品为膏霜时,其制备方法包括以下步骤:(a)在灭菌后的菠萝提取液组合物中加入水相成分,加热搅拌得水相混合物;(b)将油相组分混合,加热溶解后得油相混合物;(c)将步骤(b)制得的油相混合物加入到步骤(a)制备的水相混合物中,均质得所述美容产品。
- 根据权利要求12所述的美容产品,其特征在于:所述步骤(a)中,所述水相成分包括植物油和抗菌剂;和/或所述步骤(b)中,所述油相成分包括植物油和乳化剂;优选地,所述植物油选自以下一种或多种:甘油、青刺果油、橄榄油、牡丹籽油、澳洲坚果油、巴巴苏子油、椿花油;优选地,所述抗菌剂选自以下一种或多种:己二醇、羟苯甲酯、羟苯丙酯、苯氧乙醇;和/或优选地,所述乳化剂选自以下一种或多种:小麦乳化剂、聚甘油-3二异硬脂酸酯、二聚羟基硬脂酸、耶油基葡糖苷。
- 根据权利要求12或13所述的美容产品,其特征在于:所述步骤(a)中,所述灭菌后的菠萝提取液组合物、植物油和抗菌剂的比例为80~95:1~10:0.01~1g/g/g,优选为89~93:2~5:0.05~0.2g/g/g;所述步骤(b)中,所述植物油和乳化剂的比例为1~10:2~15g/g,优选为2~5:5~10g/g;和/或所述步骤(c)中,所述油相混合物和所述水相混合物的比例为2~25:0.5~5g/g,优选为5~10:1~2g/g;优选地,所述步骤(a)、所述步骤(b)和所述步骤(c)中,加热温度为70~120℃,更优选为70~100℃,进一步优选为80~90℃;和/或优选地,所述步骤(c)中,所述均质的时间为15~45min,更优选为20~40min,进一步优选为25~35min。
- 一种用于抗衰老的方法,其特征在于,所述方法包括:对有需要的受试者给予:权利要求1或2所述的菠萝提取液组合物;按照权利要求3至9中任一项所述的方法制备的菠萝提取液组合物;或权利要求11至14中任一项所述的美容产品。
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CN111067851A (zh) * | 2020-01-17 | 2020-04-28 | 云南白药集团股份有限公司 | 一种松茸面膜液的制备方法 |
CN111110615A (zh) * | 2020-01-17 | 2020-05-08 | 云南白药集团股份有限公司 | 一种松茸面膜液、其制备方法及应用 |
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 |
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 |
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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 |
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