CN110731927A - Enhanced whitening cosmetic composition - Google Patents
Enhanced whitening cosmetic composition Download PDFInfo
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- CN110731927A CN110731927A CN201910694704.6A CN201910694704A CN110731927A CN 110731927 A CN110731927 A CN 110731927A CN 201910694704 A CN201910694704 A CN 201910694704A CN 110731927 A CN110731927 A CN 110731927A
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- birch juice
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- ascorbyl
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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]
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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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
- A61K8/44—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof
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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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/4973—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom
- A61K8/498—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom having 6-membered rings or their condensed derivatives, e.g. coumarin
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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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/60—Sugars; Derivatives thereof
- A61K8/602—Glycosides, e.g. rutin
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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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/67—Vitamins
- A61K8/673—Vitamin B group
- A61K8/675—Vitamin B3 or vitamin B3 active, e.g. nicotinamide, nicotinic acid, nicotinyl aldehyde
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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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/67—Vitamins
- A61K8/676—Ascorbic acid, i.e. vitamin C
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- 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
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- 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
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
Abstract
The present invention provides enhanced whitening cosmetic compositions comprising (a) concentrated birch juice, and (B) or more substances selected from arbutin, tranexamic acid, niacinamide, glabridin, ascorbic acid, ascorbyl glucoside, ascorbyl dipalmitate, and ascorbyl tetraisopalmitate, wherein the concentration of the concentrated birch juice is about 1.05-6 times, preferably about 1.1-5 times, more preferably about 1.2-4 times.
Description
Technical Field
The present invention relates to enhanced whitening cosmetic compositions comprising (A) concentrated birch juice and (B) or more substances selected from arbutin, tranexamic acid, niacinamide, glabridin, ascorbic acid, ascorbyl glucoside, ascorbyl dipalmitate and ascorbyl tetraisopalmitate, wherein the concentration of the concentrated birch juice is about 1.05-6 times, preferably about 1.1-5 times, more preferably about 1.2-4 times.
Background
Birch is a deciduous tree of the betulinaceae family, and currently, there are about 100 varieties worldwide, mainly distributed in the northern temperate zone and the cold temperate zone. Wherein, there are about 29 varieties in China, and the varieties are mainly distributed in the northeast, northwest and southwest. Birch trees are mostly grown in remote mountainous areas with less human intervention and no industrial pollution. Birch sap (also called birch sap) is fresh sap obtained by cutting bark or drilling trunk of birch, and is colorless or yellowish, has no precipitate or impurity, and has light birch fragrance. The birch juice contains abundant saccharide, amino acids, vitamins, biotin, cytokinin, trace mineral elements, aromatic oil, betulin, saponin, etc., and has good skin caring effects such as moisturizing and whitening effects.
Disclosure of Invention
, the present invention relates to the use of (A) concentrated birch juice with (B) or more selected from arbutin, tranexamic acid, niacinamide, glabridin, ascorbic acid, ascorbyl glucoside, ascorbyl dipalmitate and ascorbyl tetraisopalmitate in a whitening cosmetic composition, wherein the concentration of the concentrated birch juice is about 1.05-6 times, preferably about 1.1-5 times, more preferably about 1.2-4 times.
In another aspect, the present invention provides enhanced whitening cosmetic compositions comprising (a) concentrated birch juice, and (B) or more substances selected from arbutin, tranexamic acid, niacinamide, glabridin, ascorbic acid, ascorbyl glucoside, ascorbyl dipalmitate, and ascorbyl tetraisopalmitate, wherein the concentrated birch juice is concentrated about 1.05-6 times, preferably about 1.1-5 times, more preferably about 1.2-4 times.
Unexpectedly, the inventor finds that compared with the single use of concentrated birch juice, arbutin, tranexamic acid, nicotinamide, glabridin, ascorbic acid, ascorbyl glucoside, ascorbyl dipalmitate or ascorbyl tetraisopalmitate, the combination of concentrated birch juice and arbutin, tranexamic acid, nicotinamide, glabridin, ascorbic acid, ascorbyl glucoside, ascorbyl dipalmitate and/or ascorbyl tetraisopalmitate has a remarkably better whitening effect which is far higher than the function superposition effect of the two, and the combination shows that the combination can inhibit the synthesis and the activity of tyrosinase, inhibit the synthesis and the transportation of melanin, reduce dark skin color and make the skin more white and bright, which shows that a synergistic effect is generated between the concentrated birch juice and the arbutin, the tranexamic acid, the nicotinamide, the ascorbyl dipalmitate and/or the ascorbyl tetraisopalmitat.
Birch juices involved in the present invention are obtained from Betula genus of betulinaceae family, which may be from four varieties of white birch (Betula alba), Betula luminifera (Betula pubescens), Betula pendula (Betula pendula) and birch asiana (Betula platyphylla). The birch juice is colorless, transparent, precipitate-free and impurity-free juice which is obtained by manually drilling and collecting at the base of a trunk of the birch between thawing and early spring leaf emergence and has birch faint scent and rich nutrition. The birch juice is commercially available and used as such, for example from greater Khingan over wild berry development, LLC.
The concentrated birch sap of the present invention is obtained by concentrating the above-mentioned commercially available products. Concentration methods are known in the art, such as heat concentration, low temperature vacuum concentration, membrane concentration, and the like. In the present invention, the concentration is preferably performed by a low-temperature freeze concentration or membrane concentration process, for example, commercially available birch juice stock solution is fed into a low-temperature drying device, cooled to-40 ℃ to-70 ℃, and vacuumized to 0.1 to 30Pa to perform low-temperature vacuum concentration, so as to obtain concentrated birch juice with different concentration times.
, the inventor has also found that the whitening efficacy of the concentrated birch juice is not simply linear with its concentration, but rather it shows a tendency to increase and then decrease as the concentration factor increases, therefore, it is critical to control the concentration factor of the birch juice, which in the present invention is about 1.05-6 times, preferably about 1.1-5 times, and more preferably about 1.2-4 times.
The concentrated birch juice is contained in an amount of about 10-98% by weight, preferably about 20-98% by weight, more preferably about 30-97% by weight, based on the total weight of the whitening cosmetic composition.
The component (B), arbutin, tranexamic acid, niacinamide, glabridin, ascorbic acid, ascorbyl glucoside, ascorbyl dipalmitate or ascorbyl tetraisopalmitate, are known in the art, and they are commercially available as such and used in the present invention.
The total content of the component (B) is about 0.0005 to 30% by weight, preferably about 0.001 to 10% by weight, more preferably about 0.1 to 5% by weight, most preferably about 0.5 to 3% by weight, based on the total weight of the whitening cosmetic composition.
The whitening cosmetic composition does not contain any added water, but does not exclude moisture inherently contained in the components.
Preferably, the whitening cosmetic composition does not contain a chelating agent such as EDTA salt, sodium polyphosphate, sodium metaphosphate, gluconic acid, and the like.
The whitening cosmetic composition may optionally include, in addition to the above-mentioned components (a) and (B), component (C), an ingredient commonly used in skin care cosmetic compositions. Examples of the component (C) include, but are not limited to, vehicles, active ingredients, and adjuvants, and the like. These ingredients are known in the art, and the type and amount of component (C) may be selected by those skilled in the art as desired, for example, the content of component (C) is generally about 0 to 70% by weight based on the total weight of the whitening cosmetic composition.
The vehicle includes, for example, diluents, dispersants or carriers and the like, examples of which include, but are not limited to, ethanol, dipropylene glycol, butylene glycol, and the like. The amount of said vehicle in said cosmetic composition is known in the art, and is for example generally comprised between 0.5 and 20% of the total weight of component (C).
Such active ingredients include, for example, emollients, humectants, whitening actives and the like.
Examples of such emollients include, but are not limited to, olive oil, macadamia nut oil, sweet almond oil, grape seed oil, avocado oil, corn oil, sesame oil, soybean oil, peanut oil, meadowfoam seed oil, safflower seed oil, rosa canina oil, argan oil, jojoba seed oil, sunflower seed oil, palm kernel oil, squalane, ethylhexyl palmitate, isopropyl myristate, hydrogenated polyisobutene, isohexadecane, isododecane, diethylhexyl carbonate, dioctyl carbonate, isopropyl lauroyl sarcosinate, isononyl isononanoate, hydrogenated polydecene, tris (ethylhexanoate), cetyl ethylhexanoate, bis-diethoxydiol cyclohexane 1, 4-dicarboxylate, caprylic/capric triglyceride, oleyl erucate myristate, octyldodecanol, dimethicone, octylmethicone, cetyl dimethicone, cyclopentadecyl dimethicone, and the like, examples of solid emollients include, but are not limited to, stearyl alcohol, cetyl stearyl alcohol, shark alcohol, behenyl wax, cetearyl wax.
Examples of such moisturizers include, but are not limited to, or more of glycerin, diglycerin, butylene glycol, propylene glycol, 1, 3-propanediol, dipropylene glycol, 1, 2-pentanediol, polyethylene glycol-8, polyethylene glycol-32, methyl gluceth-10, methyl gluceth-20, PEG/PPG-17/6 copolymer, glyceryl polyether-7, glyceryl polyether-26, glyceryl glucoside, PPG-10 methyl glucose ether, PPG-20 methyl glucose ether, PEG/PPG/polytetramethylene glycol-8/5/3 glycerin, sucrose, trehalose, rhamnose, mannose, raffinose, betaine, erythritol, xylitol, urea, glyceryl polyether-5 lactate, sodium hyaluronate, hydrolyzed sodium hyaluronate, acetylated sodium hyaluronate, sodium polyglutamate, hydrolyzed sclerotium rolum, pullulan, tremella polysaccharides, tamarind seed polysaccharides, and the like.
The whitening active ingredients, including but not limited to of one or more of kojic acid, ascorbyl glucoside, arbutin, tranexamic acid, niacinamide, phytosterol/behenyl alcohol/octyldecanol lauroyl glutamate, phenethyl resorcinol, turmeric root extract, birch bark extract, ceramide 2, ceramide 3, acetyl phytosphingosine, resveratrol, bark extract of Pterocarpus marsupium, root extract of Coleus forskohlii, pepper seed extract, ubiquinone, cholesterol stearate, ascorbic acid, ascorbyl dipalmitate, tocopherol (vitamin E), tocopheryl acetate, bisabolol, tetraisopalmitate ascorbate, pyridoxine dicaprylate, pyridoxine dipalmitate, retinol palmitate, phytosterol/octyldodecanoyl glutamate, bis-behenyl alcohol/isostearyl alcohol/phytosterol dimer linoleate, phytosteryl dioleate, phytosteryl macadamia oleate, various peptides, various plant extracts, etc. are generally present in the cosmetic composition at a level of 0.01, for example.
Such adjuvants include, for example, emulsifiers, thickeners, preservatives, fragrances and the like.
Examples of such emulsifiers include, but are not limited to, cetearyl olivate, sorbitan olivate, polysorbate-60, polysorbate-80, methylgluco-sesquistearate, PEG-20 methylgluco-sesquistearate, PEG-40 hydrogenated castor oil, PPG-26-butanoeth-26, PEG-4 polyglyceryl-2 stearate, PEG-60 hydrogenated castor oil, steareth-2, steareth-21, PPG-13-decyltetraeth-24, cetearyl glucoside, PEG-100 stearate, glyceryl stearate SE, coco glucoside, ceteareth-25, PEG-40 stearate, polyglyceryl-3 methylgluco distearate, glyceryl stearate citrate, polyglyceryl-10 stearate, polyglyceryl-10 myristate, polyglyceryl-10 dioleate, polyglyceryl-10 laurate, polyglyceryl-10 isostearate, polyglyceryl-10 oleate, polyglyceryl-10 diisostearate, polyglyceryl-6 laurate, polyglyceryl-6 myristate, sucrose % of the various cosmetic components generally known in the art (e.g. 0.32% of the total weight of such emulsifiers, e.g. the cosmetic components are included in the cosmetic composition.
Examples of the thickener include, but are not limited to, kinds or more of high molecular polymers such as carbomers, acrylates and derivatives thereof, xanthan gum, arabic gum, polyethylene glycol-14M, polyethylene glycol-90M, succinoglycan, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, etc. the content of the thickener in the cosmetic composition is known in the art, and for example, it is usually 0.1 to 10% by weight based on the total weight of the component (C).
Examples of such preservatives include, but are not limited to, of methyl paraben, ethyl paraben, propyl paraben, phenoxyethanol, benzyl alcohol, phenyl ethanol, bis (hydroxymethyl) imidazolidinyl urea, potassium sorbate, sodium benzoate, chlorphenesin, sodium dehydroacetate, and the like, the level of such preservatives in the cosmetic composition is known in the art, e.g., it is typically 0.01 to 30% by weight of component (C).
The whitening cosmetic composition of the present invention may be prepared by any suitable method known in the art. For example, it is prepared using a dissolving tank, an emulsifying pot, a disperser, a transfer pump, etc., which are commonly used in the cosmetic field. The preparation method comprises putting water soluble substance into water phase dissolving kettle, putting oil soluble substance into oil phase dissolving kettle, heating the two kettles to about 80 deg.C, wherein the raw material easy to agglomerate can be pre-dispersed with disperser. After the dissolution is finished, the oil phase and the water phase are conveyed into an emulsifying pot, and homogenized and emulsified for about 5-15 minutes. After emulsification is finished, the temperature of the material body is reduced to normal temperature, optional essence, preservative and the like are added, and the pH of the product is adjusted according to needs. After the relevant detection indexes are qualified, the products can be filled and delivered.
The preparation method can be deleted or adjusted according to the requirements of dosage forms. The whitening cosmetic composition may be formulated into various forms such as cream, milky lotion, essence, etc., as required.
Examples
The invention is described in further detail at with reference to the following examples, however, it should be understood that these examples and comparative examples are intended only to illustrate the invention in more detail and should not be construed as limiting in any way the scope of the appended claims.
Example 1: influence on expression of genes related to melanin synthesis and transportation
In this example, the effects of birch juice and arbutin, tranexamic acid, and combinations thereof on melanin synthesis and transport-related gene expression at different concentration ratios were tested and compared.
1. Concentration of birch sap
Fresh birch juice stock solution purchased from Daxingan mountain surpassing wild berry development Limited company is input into a low-temperature drying device, cooled to-65 ℃, vacuumized to 0.1Pa, and concentrated to 2 times and 8 times respectively.
2. Testing
An experimental instrument: a fluorescent quantitative PCR instrument (Roche), a super clean bench (Sujing), a carbon dioxide incubator (Binder), a microplate reader (BIO-TEK) and a micro oscillator.
Experimental reagents and consumables: human primary melanocytes, 6-well plates, melanocyte culture solution, RNA extraction kit, reverse transcription kit, Trizol lysate and the like.
The sample loading information comprises that the loading amount of the single raw material is parts of the whole raw material, and the compound raw material is half of the birch juice raw material and half of 0.1 percent of other active raw materials.
Melanocyte-based gene expression analysis procedures were as follows:
(1) inoculation: cells were seeded into 6-well plates at a seeding density of 5E 5/well at 37 ℃ and 5% CO2Incubating in an incubator overnight;
(2) administration: when the cell plating rate in the 6-hole plate reaches about 60%, adding the test substances of each group, and arranging 6 multiple holes in each group;
(3) collecting a sample: at 37 ℃ and 5% CO2After 24 hours in the incubator, discarding the culture solution, adding 1mL of Trizol into each hole, blowing and beating the lysed cells, and collecting samples;
(4) and (3) PCR detection: extracting RNA, carrying out reverse transcription to cDNA, and carrying out fluorescent quantitative PCR detection;
(5) and (3) analysis: by using 2-△△CTThe method carries out result calculation and adopts a T-Test method for statistical analysis.
The test results are shown in the following table:
the results show that in the aspect of inhibiting melanin synthesis and transferring gene transcription, the effect of the combination of the 2-time concentrated birch juice and the arbutin or the tranexamic acid is obviously better than that of the combination of the single birch juice stock solution, the 2-time concentrated birch juice, the arbutin and the tranexamic acid, and simultaneously, the combination of the birch juice stock solution and the arbutin or the tranexamic acid, and the transcription of melanin synthesis related genes TYR and PMEL17 and melanin transfer related genes Rab-27a and MyosinVa is obviously inhibited. When the birch juice is concentrated to 8 times, the effect of combining the birch juice with other active matters is obviously weakened, even the effect of combining the stock solution birch juice with other active matter raw materials is not as good.
Example 2: influence on expression of melanin synthesis and transport related protein
In this example, the effect of birch juice and its niacinamide, glabridin, and combinations thereof at different fold concentrations on the expression of proteins associated with melanin synthesis and transport was tested and compared.
1. Concentration of birch sap
Fresh birch juice stock solution purchased from Daxingan mountain surpassing wild berry development Limited company is input into a low-temperature drying device, cooled to-65 ℃, vacuumized to 0.1Pa, and concentrated to 2 times and 8 times respectively.
2. Testing
An experimental instrument: super clean bench (Sujing), plate washing machine (BIO-RAD), enzyme labeling instrument (BIO-TEK), carbon dioxide incubator (Binder)
Experimental reagents and consumables: human primary melanocytes, 12-hole plates, melanocyte culture solution, ELISA detection kits with different indexes and the like.
The sample loading information comprises that the loading amount of the single raw material is parts of the whole raw material, and the compound raw material is half of the birch juice raw material and half of 0.1 percent of other active raw materials.
The test procedure was as follows:
(1) inoculation: inoculating at 2E 5/well in 12-well culture plate at 37 deg.C and 5% CO2Culturing in an incubator, and changing the culture medium for times every two days;
(2) administration: when the cell fusion reaches more than 60 percent again, adding the test substances of different groups, and arranging 6 compound holes in each group;
(3) collecting a sample: at 37 ℃ and 5% CO2After 48 hours in the incubator, discarding the culture solution, adding 1mL of Trizol into each hole, blowing and beating the lysed cells, and collecting the samples;
(4) and (3) detection: performing index measurement according to a measurement method of the ELISA kit;
(5) and (3) analysis: and carrying out statistical analysis by adopting a T-Test method.
The test results are shown in the following table.
The results show that the effect of the combination of the 2-time concentrated birch juice and the nicotinamide or the glabridin is obviously better than that of the single combination of the birch juice stock solution, the 2-time concentrated birch juice, the nicotinamide and the glabridin in the aspect of inhibiting the melanin synthesis and the expression of the transport protein, and the expression levels of the melanin synthesis related proteins TYR, TYR1, TYR2 and the melanin transport related protein Rab-27a are all obviously reduced. When the birch juice is concentrated to 8 times, the effect of combining the birch juice with other active matters is obviously weakened, even the effect of combining the stock solution birch juice with other active matter raw materials is not as good.
Example 3: effect on melanogenesis in melanocytes
In this example, the effect of birch juice and its ascorbic acid, ascorbyl glucoside, ascorbyl dipalmitate or ascorbyl tetraisopalmitate, and combinations thereof, at different concentrations on melanin production in melanocytes was tested and compared.
1. Concentration of birch sap
Fresh birch juice stock solution purchased from Daxingan mountain surpassing wild berry development Limited company is input into a low-temperature drying device, cooled to-65 ℃, vacuumized to 0.1Pa, and concentrated to 2 times and 8 times respectively.
2. Testing
An experimental instrument: clean bench (Sujing), carbon dioxide incubator (Binder), water bath, enzyme labeling instrument (BIO-TEK).
Experimental reagents and consumables: human primary melanocytes, 12-well plates, melanocyte culture solution and NaOH lysate.
The sample loading information comprises that the loading amount of the single raw material is parts of the whole raw material, and the compound raw material is half of the birch juice raw material and half of 0.1 percent of other active raw materials.
The experimental method comprises the following steps:
(1) inoculation: inoculating at 2E 5/well density in 12-well culture plate at 37 deg.C and 5% CO2The culture box was used for culturing, and the culture medium was changed times every two days.
(2) Administration: when the cell fusion reaches more than 60% again, adding the test substances of different groups, and simultaneously setting a negative control group (without adding drugs) and setting 6 multiple holes in each group.
(3) Collecting a sample: 37 ℃ and 5% CO2After 48 hours in the incubator, the culture medium was discarded, washed with PBS solution 3 times, and then 100. mu.l of 1mol/L NaOH was added thereto, and the mixture was subjected to 80 ℃ water bath for 1 hour.
(4) And (3) detection: after completion of the water bath, centrifugation was carried out at 10000rpm for 10 minutes, and the supernatant was aspirated and the absorbance (A) was measured at 460nm with a microplate reader, and the melanin inhibition was calculated according to the following formula: melanin inhibition (%) was 100% (a negative control group-a administration group)/a negative control group%
(5) And (3) analysis: statistical analysis was performed using the T-Test method.
Sample (I) | Melanin inhibition% |
Stock solution of birch juice | 10.6%±1.2% |
2-fold concentrated birchJuice product | 23.6%±2.2% |
8 times concentrated birch juice | 12.3%±1.3% |
Ascorbic acid | 3.7%±1.7% |
Ascorbic acid glucoside | 5.3%±1.9% |
Ascorbic acid dipalmitate | 8.2%±1.2% |
Ascorbyl tetraisopalmitate | 3.4%±1.1% |
Birch juice stock solution and ascorbic acid | 16.4%±2.2% |
Birch juice stock solution and ascorbic acid glucoside | 17.7%±1.9% |
Birch juice stock solution and ascorbyl dipalmitate | 15.3%±1.4% |
Stock solution of birch juice and ascorbyl tetraisopalmitate | 15.9%±1.6% |
2 times of concentrated birch juice stock solution and ascorbic acid | 25.7%±1.5% |
2 times concentrated birch juice stock solution and ascorbic acid glucoside | 28.2%±2.3% |
2 times of concentrated birch juice stock solution and ascorbyl dipalmitate | 27.6%±1.2% |
2 times concentrated birch juice stock solution and ascorbic acid tetraisopalmitate | 24.8%±1.6% |
8 times concentrated birch juice stock solution and ascorbic acid | 14.6%±1.2% |
8 times concentrated birch juice stock solution and ascorbic acid glucoside | 15.1%±1.5% |
8 times of concentrated birch juice stock solution and ascorbyl dipalmitate | 13.9%±1.8% |
8 times concentrated birch juice stock solution and ascorbic acid tetraisopalmitate | 15.4%±0.9% |
The above results indicate that the effect of using the combination of the 2-fold concentrated birch juice and ascorbic acid, ascorbyl glucoside, ascorbyl dipalmitate or ascorbyl tetraisopalmitate is significantly better than that of using the combination of the birch juice stock solution, the 2-fold concentrated birch juice and ascorbic acid, ascorbyl glucoside, ascorbyl dipalmitate, ascorbyl tetraisopalmitate alone, and that the melanin production inhibition rate of the melanocytes is significantly increased than that of using the combination of the birch juice stock solution and ascorbic acid, ascorbyl glucoside, ascorbyl dipalmitate or ascorbyl tetraisopalmitate. When the birch juice is concentrated to 8 times, the effect of combining the birch juice with other active matters is obviously weakened, even the effect of combining the stock solution birch juice with other active matter raw materials is not as good.
Example 4: effect on melanogenesis in 3D melanocyte models
In this example, the effect of birch juice and its ascorbic acid, as well as their compositions, at different fold concentrations on melanin production in a 3D melanocyte model was tested and compared.
1. Concentration of birch sap
Fresh birch juice stock solution purchased from Daxingan mountain surpassing wild berry development Limited company is input into a low-temperature drying device, cooled to-65 ℃, vacuumized to 0.1Pa, and concentrated to 2 times and 8 times respectively.
2. Test method
An experimental instrument: clean bench (Sujing), carbon dioxide incubator (Binder), UVB irradiator, water bath, enzyme labeling instrument (BIO-TEK).
Experimental reagents and consumables: 3D melanoderm model (self-made in laboratory), model culture solution, NaOH lysate.
The sample loading information comprises that the loading amount of the single raw material is parts of the whole raw material, and the compound raw material is half of the birch juice raw material and half of 0.1 percent of other active raw materials.
The experimental method comprises the following steps:
(1)3D model construction: the 3D skin model was constructed using keratinocytes and melanocytes.
(2) Molding and administration: UVB irradiation treatment (UVB: 50 mJ/cm) was performed every day when the model was shipped from the factory, i.e., 0 day2) Then, quantitative samples were applied to the corresponding model surfaces, and model control groups were applied with model culture solution alone, each group consisting of 6 duplicate wells, applications per day, for a total duration of 4 days.
(3) Collecting and detecting: after the reaction of the sample, the model was taken out, washed 3 times with PBS and placed in a centrifuge tube, then 100. mu.l of 1mol/L NaOH was added, water bath at 80 ℃ was carried out for 1 hour, after the water bath was completed, centrifugation was carried out at 10000rpm for 10 minutes, the supernatant was aspirated and the absorbance (A) was measured at 460nm with a microplate reader, and the melanin inhibition was calculated according to the following formula:
melanin inhibition (%) was (a model control group-a administration group)/a model control group x 100%
(4) And (3) analysis: statistical analysis was performed using the T-Test method.
The test results are shown in the following table.
Sample (I) | Melanin inhibition% |
Stock solution of birch juice | 7.7%±0.6% |
2 times concentrated birch juice | 18.3%±1.1% |
8 times concentrated birch juice | 8.5%±0.7% |
Ascorbic acid | 4.5%±0.8% |
Birch juice stock solution and ascorbic acid | 14.6%±1.2% |
2 times of concentrated birch juice stock solution and ascorbic acid | 24.2%±1.8% |
8 times concentrated birch juice stock solution and ascorbic acid | 12.9%±1.3% |
The results show that the effect of the combination of the 2-time concentrated birch juice and the ascorbic acid is obviously better than that of the single birch juice stock solution, the 2-time concentrated birch juice and the ascorbic acid in a 3D skin model, and simultaneously the composition of the birch juice stock solution and the ascorbic acid is also obviously better than that of the composition of the birch juice stock solution and the ascorbic acid, and the melanin inhibition rate of the 3D skin model is obviously improved. When the birch juice is concentrated to 8 times, the effect of combining the birch juice with other active matters is obviously weakened, even the effect of combining the stock solution birch juice with other active matter raw materials is not as good.
Example 5: preparation of whitening essence emulsion composition
The formula of the whitening essence emulsion composition is shown in the following table:
the whitening essence composition is prepared as follows:
1, A, B phases are respectively heated to 80 ℃ to be dissolved uniformly;
2. keeping the temperature at 80 ℃, slowly adding the phase B into the phase A under stirring, stirring for 10 minutes, and homogenizing at 10000 rpm/minute for 5 minutes;
3. slowly stirring and cooling to 60 ℃ after defoaming, adding the pre-dispersed phase C into the phase AB, and homogenizing for 3 minutes at 10000 rpm/minute;
4. adding the phase D into the phase ABC, stirring and cooling to 40 ℃ to discharge.
Using the half-face control test method, 20 volunteers were tested before and 8 weeks after using the product as follows:
1) measuring the whiteness value L of facial skin by using a CK-skinncolor instrument;
2) the melanin index of the skin at the same location was measured using a Maxmeter MX18 reflectance spectrometer.
3) The facial patterns of the subjects were collected by VISIA-CR under different light sources at different times and the skin gloss, whiteness L and visible spot area of the subject's face were analyzed by IPP software at different test time points and measurement areas.
The results showed that 8 weeks using the whitening essence cream composition comprising a combination of 1.5 times concentrated birch juice and arbutin was able to increase the facial skin brightness L value by 5.23%, reduce the melanin content by 17.46%, increase the skin brightness by 20.37%, and significantly reduce the area of facial stains in 16 out of 20 subjects.
The technical solutions of the above-described embodiments are preferred embodiments of the present invention, and several modifications and changes can be made without departing from the principle of the present invention, and these modifications and changes should also be considered as being within the protection scope of the present invention.
Claims (6)
- Use of (a) concentrated birch juice with B) or more selected from arbutin, tranexamic acid, niacinamide, glabridin, ascorbic acid, ascorbyl glucoside, ascorbyl dipalmitate and ascorbyl tetraisopalmitate in a whitening cosmetic composition, wherein the concentrated birch juice is concentrated 1.05-6 times, preferably 1.1-5 times, more preferably 1.2-4 times.
- 2. The use of claim 1, wherein the whitening cosmetic composition does not comprise any added water.
- whitening cosmetic composition comprising (A) concentrated birch juice and (B) or more substances selected from arbutin, tranexamic acid, niacinamide, glabridin, ascorbic acid, ascorbyl glucoside, ascorbyl dipalmitate and ascorbyl tetraisopalmitate, wherein the concentrated birch juice is concentrated 1.05-6 times, preferably 1.1-5 times, more preferably 1.2-4 times.
- 4. The whitening cosmetic composition of claim 3, wherein the whitening cosmetic composition does not include any added water.
- 5. The whitening cosmetic composition of claim 3 or 4, wherein the concentrated birch juice of the component (A) is contained in an amount of 10 to 98% by weight, preferably 20 to 98% by weight, more preferably 30 to 97% by weight, based on the total weight of the whitening cosmetic composition.
- 6. The whitening cosmetic composition of any of claims 3-5, wherein the component (B) is present in an amount of 0.0005 to 30% by weight, preferably about 0.001 to 10% by weight, more preferably about 0.1 to 5% by weight, most preferably about 0.5 to 3% by weight, based on the total weight of the whitening cosmetic composition.
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CN112263496A (en) * | 2020-10-26 | 2021-01-26 | 广州环亚化妆品科技有限公司 | Whitening agent, whitening oil gel containing whitening agent and preparation method of whitening oil gel |
WO2021017835A1 (en) * | 2019-07-30 | 2021-02-04 | 浙江养生堂天然药物研究所有限公司 | Enhanced whitening cosmetic composition |
WO2021017911A1 (en) * | 2019-07-30 | 2021-02-04 | 浙江养生堂天然药物研究所有限公司 | Enhanced hair loss prevention and hair growth promotion composition |
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CN113975215A (en) * | 2021-11-18 | 2022-01-28 | 广州梵之容化妆品有限公司 | Whitening cream and preparation method thereof |
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