CN110664718B - Whitening skin care product - Google Patents
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- CN110664718B CN110664718B CN201910966610.XA CN201910966610A CN110664718B CN 110664718 B CN110664718 B CN 110664718B CN 201910966610 A CN201910966610 A CN 201910966610A CN 110664718 B CN110664718 B CN 110664718B
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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
- A61K8/68—Sphingolipids, e.g. ceramides, cerebrosides, gangliosides
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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
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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
- 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
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/981—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of mammals or bird
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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
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/59—Mixtures
- A61K2800/592—Mixtures of compounds complementing their respective functions
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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 invention relates to a whitening skin care product, which comprises N-acetylneuraminic acid, and also comprises one or more compositions of B-phase substances, namely nicotinamide, arbutin, vitamin C and cubilose extract, wherein the proportion of the N-acetylneuraminic acid to the B phase is 1: 1-4, the whitening skin care product contains N-acetylneuraminic acid, has an obvious inhibition effect on tyrosinase, can effectively prevent the generation of melanin, and is safe and effective when being used on cells.
Description
Technical Field
The invention belongs to the field of daily chemical products, and particularly relates to a whitening skin care product.
Background
The skin is the biggest organ of human body, and direct with external environment contact, has effects such as protection, excretion, regulation body temperature and feeling external stimulus, and facial skin has the effect of promoting the outward appearance more concurrently. With the improvement of living standard of people, people pay more and more attention to their skin, and some efficacy products are more and more favored, and especially, whitening skin care products are almost the goals pursued by all women. The whitening products in the current market are mixed, most products with whitening effects are not in compliance, the phenomena of heavy metal exceeding standard, fluorescent agent exceeding standard and the like often occur, and the safe whitening effect cannot be achieved.
In melanocytes, tyrosine is oxidized into dopa and dopaquinone in sequence under the catalytic action of tyrosinase, and the dopaquinone is further converted into dopachrome; under the respective catalytic actions of DHI enzyme, dopachrome tautomerase and DHICA oxidase, dopachrome gradually forms melanin. Tyrosinase is therefore the first rate-limiting and key enzyme in melanin synthesis.
The existing whitening products are difficult to effectively inhibit tyrosinase, so that the whitening effect is poor, and a whitening skin care product capable of effectively inhibiting tyrosinase is needed.
Disclosure of Invention
In order to solve the problems, the invention provides a whitening skin care product.
A whitening skin care product comprises N-acetylneuraminic acid.
Further, the whitening skin care product also comprises one or more compositions of B-phase substances, namely nicotinamide, arbutin, vitamin C and cubilose extract;
the ratio of the N-acetylneuraminic acid to the B phase is 1:1 to 4.
Further, the phase B is any one of niacinamide, arbutin, vitamin C, bird's nest extract, a combination of niacinamide and arbutin, a combination of niacinamide and bird's nest extract and a combination of niacinamide, arbutin and bird's nest extract.
Further, in the combination of niacinamide and arbutin: the ratio of the nicotinamide to the arbutin is 1:0.6 to 1.2;
in the combination of niacinamide and bird's nest extract: the ratio of the nicotinamide to the cubilose extract is as follows: 1: 0.8-1.2;
the combination of the niacinamide, the arbutin and the cubilose extract comprises the following components: the proportion of the nicotinamide, the arbutin and the cubilose extract is as follows: 1: 0.6-1: 0.8-1.2.
Furthermore, the skin care product also comprises a humectant and/or an auxiliary material acceptable in the field of skin care products.
Further, the ratio of the N-acetylneuraminic acid to the humectant is 0.1-3: 1.
Further, the humectant comprises one or more of butylene glycol, hexylene glycol, squalane, hyaluronic acid, panthenol, beta-glucan, and sodium polyglutamate.
Further, the humectant is a composition of squalane, hyaluronic acid, panthenol, beta-glucan and sodium polyglutamate, and the composition comprises 10-20% of squalane, 10-20% of hyaluronic acid, 20-40% of panthenol, 20-40% of beta-glucan and 20-40% of sodium polyglutamate.
Furthermore, the auxiliary materials acceptable in the field of skin care products comprise one or more of solubilizer, preservative, antioxidant, pH regulator, penetration enhancer, liposome, humectant, thickener, chelating agent, skin feel regulator, surfactant, emulsifier, essence and pigment.
Further, the skin care product is in the form of cream, emulsion, solution, film, aerosol or spray.
The whitening skin care product contains N-acetylneuraminic acid, has an obvious inhibition effect on tyrosinase, can effectively prevent the generation of melanin, and is safe and effective when the N-acetylneuraminic acid acts on cells. Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by the practice of the invention. The objectives and other advantages of the invention will be realized and attained by the structure particularly pointed out in the written description and claims hereof as well as the appended drawings.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and those skilled in the art can also obtain other drawings according to the drawings without creative efforts.
FIG. 1 is a graph showing the results of the inhibition experiment of tyrosinase by the groups of N-acetylneuraminic acid, nicotinamide, arbutin and bird's nest extract A7 at different concentrations according to the example of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention provides a whitening skin care product, which comprises N-acetylneuraminic acid.
The whitening skin care product also comprises one or more compositions of B-phase substances of nicotinamide, arbutin, vitamin C and cubilose extract;
the ratio of the N-acetylneuraminic acid to the B-phase substance is 1: 1-4, the whitening effect of the whitening skin care product can be enhanced through the matching of the N-acetylneuraminic acid and the phase B substance.
The phase B substance is any one of nicotinamide, arbutin, vitamin C, bird's nest extract, combination of nicotinamide and arbutin, combination of nicotinamide and bird's nest extract and combination of nicotinamide, arbutin and bird's nest extract;
preferably, the B phase substance is nicotinamide combined with arbutin and nidus Collocaliae extract.
In the combination of niacinamide and arbutin: the ratio of nicotinamide to arbutin is 1:0.6 to 1.2;
in the combination of amide and bird's nest extract: the proportion of the nicotinamide to the cubilose extract is as follows: 1: 0.8-1.2;
in the combination of niacinamide, arbutin and bird's nest extract: the proportion of the niacinamide, the arbutin and the cubilose extract is as follows: 1: 0.6-1: 0.8-1.2.
The whitening skin care product also comprises a humectant and/or an auxiliary material acceptable in the field of skin care products, wherein the humectant and/or the auxiliary material acceptable in the field of skin care products can ensure the stability and long-term effectiveness of N-acetylneuraminic acid and a B-phase substance in the whitening skin care product, and is beneficial to improving the skin feeling experience of the whitening skin care product.
In the whitening skin care product, the ratio of the N-acetylneuraminic acid to the humectant is 0.3-1: 1.
The humectant comprises one or more of butanediol, hexanediol, squalane, hyaluronic acid, panthenol, beta-glucan and sodium polyglutamate.
The humectant is preferably a composition of squalane, hyaluronic acid, panthenol, beta-glucan and sodium polyglutamate, and the humectant contains 10-20% of squalane, 10-20% of hyaluronic acid, 20-40% of panthenol, 20-40% of beta-glucan and 20-40% of sodium polyglutamate.
The acceptable auxiliary materials in the field of skin care products comprise one or more of solubilizer, preservative, antioxidant, pH regulator, penetration enhancer, liposome, humectant, thickener, chelating agent, skin feeling regulator, surfactant, emulsifier, essence and pigment.
The skin care product of the invention is in the form of cream, emulsion, solution, film, aerosol or spray.
The invention will be further illustrated with reference to specific examples, which are to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever. Test methods in which specific conditions are not specified in the following examples are generally carried out under conventional conditions or under conditions recommended by the manufacturer. All percentages and parts are by weight unless otherwise indicated.
Example 1 preparation of whitening skin Care products
The whitening component is N-acetylneuraminic acid.
The whitening skin care product comprises the following components in percentage by weight: 3% of N-acetylneuraminic acid, 3% of a humectant, 0.5% of a preservative, 4% of a thickening agent, 0.02% of a chelating agent, the pH value is adjusted to 6.0, and the balance is water.
The humectant is prepared from butanediol, hexanediol and hyaluronic acid according to a mass ratio of 9: 4: 2, the preservative is methyl hydroxybenzoate, the thickening agent is xanthan gum, the chelating agent is disodium ethylene diamine tetraacetate, and the pH regulator is triethanolamine.
Example 2 preparation of whitening skin Care products
The whitening component is a combination of N-acetylneuraminic acid and nicotinamide.
The whitening skin care product comprises the following components in percentage by weight: 2% of N-acetylneuraminic acid, 1% of B-phase nicotinamide composition, 3% of a humectant, 0.5% of a preservative, 4% of a thickening agent, 0.02% of a chelating agent, and the balance of water, wherein the pH value is adjusted to 6.0.
The humectant is prepared from butanediol, hexanediol and hyaluronic acid according to a mass ratio of 9: 4: 2, the preservative is methyl hydroxybenzoate, the thickening agent is xanthan gum, the chelating agent is disodium ethylene diamine tetraacetate, and the pH regulator is triethanolamine.
Example 3 preparation of whitening skin Care products
The whitening component is a combination of N-acetylneuraminic acid and arbutin.
The whitening skin care product comprises the following components in percentage by weight: 0.8% of N-acetylneuraminic acid, 1% of B-phase arbutin, 3% of humectant, 0.5% of preservative, 4% of thickening agent, 0.02% of chelating agent, pH adjusted to 6.0 and the balance of water.
The humectant is prepared from butanediol, hexanediol and hyaluronic acid according to a mass ratio of 9: 4: 2, the preservative is methyl hydroxybenzoate, the thickening agent is xanthan gum, the chelating agent is disodium ethylene diamine tetraacetate, and the pH regulator is triethanolamine.
Example 4 preparation of whitening skin Care products
The whitening component is a combination of N-acetylneuraminic acid and vitamin C.
The whitening skin care product comprises the following components in percentage by weight: 0.8% of phase A N-acetylneuraminic acid, 1.1% of B vitamin C, 4% of humectant, 0.5% of preservative, 4% of thickening agent, 0.02% of chelating agent, pH adjusted to 6.0 and the balance of water.
The humectant is prepared from butanediol, hexanediol and hyaluronic acid according to a mass ratio of 9: 4: 2, the preservative is methyl hydroxybenzoate, the thickening agent is xanthan gum, the chelating agent is disodium ethylene diamine tetraacetate, and the pH regulator is triethanolamine.
Example 5 preparation of whitening skin Care products
The whitening component is a combination of N-acetylneuraminic acid and an extract of bird's nest.
The whitening skin care product comprises the following components in percentage by weight: 1% of phase A N-acetylneuraminic acid, 2% of phase B cubilose extract, 3% of humectant, 0.5% of preservative, 4% of thickening agent, 0.02% of chelating agent, pH adjusted to 6.0 and the balance of water.
The humectant is prepared from butanediol, hexanediol and hyaluronic acid according to a mass ratio of 9: 4: 2, the preservative is methyl hydroxybenzoate, the thickening agent is xanthan gum, the chelating agent is disodium ethylene diamine tetraacetate, and the pH regulator is triethanolamine.
Example 6 preparation of whitening skin Care products
The whitening component is a combination of N-acetylneuraminic acid, nicotinamide and arbutin.
The whitening skin care product comprises the following components in percentage by weight: 1% of N-acetylneuraminic acid, 1% of nicotinamide, 1% of arbutin, 3% of humectant, 0.5% of preservative, 4% of thickening agent, 0.02% of chelating agent, pH adjusted to 6.0 and the balance of water.
The humectant is prepared from butanediol, hexanediol and hyaluronic acid according to a mass ratio of 9: 4: 2, the preservative is methyl hydroxybenzoate, the thickening agent is xanthan gum, the chelating agent is disodium ethylene diamine tetraacetate, and the pH regulator is triethanolamine.
Example 7 preparation of whitening skin Care products
The whitening component is N-acetylneuraminic acid, nicotinamide and nidus Collocaliae extract.
The whitening skin care product comprises the following components in percentage by weight: 1% of N-acetylneuraminic acid, 1% of nicotinamide, 1% of cubilose extract, 3% of humectant, 0.5% of preservative, 4% of thickening agent, 0.02% of chelating agent, pH adjusted to 6.0 and the balance of water.
The humectant is prepared from butanediol, hexanediol and hyaluronic acid according to a mass ratio of 9: 4: 2, the preservative is methyl hydroxybenzoate, the thickening agent is xanthan gum, the chelating agent is disodium ethylene diamine tetraacetate, and the pH regulator is triethanolamine.
Example 8 preparation of whitening skin Care products
The whitening component is a combination of niacinamide, arbutin and cubilose extract.
The whitening skin care product comprises the following components in percentage by weight: 1% of N-acetylneuraminic acid, 2% of a combination of nicotinamide, arbutin and cubilose extract, 3% of a humectant, 0.5% of a preservative, 4% of a thickening agent, 0.02% of a chelating agent, the pH is adjusted to 6.0, and the balance is water;
the humectant is prepared from butanediol, hexanediol and hyaluronic acid according to a mass ratio of 9: 4: 2, the preservative is methyl hydroxybenzoate, the thickening agent is xanthan gum, the chelating agent is disodium ethylene diamine tetraacetate, and the pH regulator is triethanolamine.
Skin care product test experiment
The skin care products prepared in examples 1-8 were all tested as follows:
the stability test is carried out, the materials are placed for 12 weeks at the temperature of minus 20 ℃, 4 ℃, room temperature and 48 ℃ respectively in a circulating way (four temperature conditions are changed continuously), and the color and the shape of each material are kept stable.
Secondly, skin safety test proves that the ointment has no stimulation to the skin, does not cause adverse reactions such as erythema, desquamation, stabbing pain, burning heat and the like, and can be used safely.
Using a sensory test, the evaluation result of the 20 female volunteers continuously using the prepared skin cream on the face for 8 weeks shows that after the skin cream containing the N-acetylneuraminic acid composition is used, the skin is obviously found to be white, the permeability of the skin is improved, the tender and plumpness of the skin is improved, and the skin looks younger.
Inhibition experiment of N-acetylneuraminic acid and composition thereof on tyrosinase at different concentrations
Firstly, raw material preparation:
testing raw materials: single-component N-acetylneuraminic acid is dissolved in a methanol solution to be prepared into the following concentration (%): 0.2%, 0.5%, 0.7% and 0.9%;
n-acetylneuraminic acid and nicotinamide a1 group: the N-acetylneuraminic acid and the nicotinamide are prepared according to the mass ratio of 1: 1;
n-acetylneuraminic acid and arbutin a2 group: the N-acetylneuraminic acid and the arbutin are prepared according to the mass ratio of 1: 1;
n-acetylneuraminic acid and vitamin C A3 group: the N-acetylneuraminic acid and the arbutin are prepared according to the mass ratio of 1: 1;
n-acetylneuraminic acid and cubilose extract group a 4: n-acetylneuraminic acid and the cubilose extract in a mass ratio of 1:1, preparation;
n-acetylneuraminic acid, niacinamide and arbutin group a 5: n-acetylneuraminic acid, nicotinamide and arbutin are added at a ratio of 1:1, preparation;
n-acetylneuraminic acid, nicotinamide and bird's nest extract group a 6: n-acetylneuraminic acid, nicotinamide and bird's nest extracts are mixed according to a ratio of 1:1, preparation;
n-acetylneuraminic acid, niacinamide, arbutin and bird's nest extract group a7, N-acetylneuraminic acid, niacinamide, arbutin and bird's nest extract are mixed according to a ratio of 1: 1: 1:1, preparation;
the compositions of the above groups are prepared and dissolved by methanol, and the following concentrations (%) are prepared for each group: 0.2%, 0.5%, 0.7% and 0.9%.
Coagulant salt buffer solution (PH 6.8): first, 0.2M sodium dihydrogen phosphate and 0.2M disodium hydrogen phosphate were prepared. 0.2M sodium dihydrogen phosphate: weighing 71.6g Na2HPO4-12H2O, dissolved in 1000ml of deionized water. 0.2M disodium hydrogen phosphate: weighing 31.2g of NaH2PO4-12H2O, dissolved in 1000ml of deionized water. 51ml of sodium dihydrogen phosphate and 49ml of disodium hydrogen phosphate are taken to obtain 0.2M, PH-6.8 phosphoric acid buffer solution.
Tyrosinase solution: cleaning potato, and precooling at 4 deg.C for about 4 h. Peeling, cutting into 1.0cm3D-form, and freeze at-20 ℃ overnight. Weighing, adding 4 deg.C pre-cooled sodium sulfate buffer solution at a ratio of 1:1, homogenizing with tissue triturator, filtering with 3 layers of gauze, centrifuging the filtrate at 4000r/min for 10min to obtain supernatant as obtained tyrosinase crude enzyme solution, storing at 4 deg.C, and using up in 2 h.
The first experiment method comprises the following steps:
sample group: each group of the test materials comprises 1ml of test raw materials, 1ml of phosphate buffer solution and 1ml of tyrosinase solution;
first control group: 2ml of phosphate buffer solution and 1ml of tyrosinase solution;
second control group: 3ml of phosphoric acid buffer solution;
third control group: each test sample 1ml and phosphate buffer 2ml
After mixing, the test group and the control group were incubated at room temperature (28 ℃) for 20min and absorbance detection (475nm) was rapidly performed. The tyrosinase inhibition rate was calculated using the following formula:
wherein A is the absorbance measured by the first control group, B is the absorbance measured by the second control group, C is the absorbance measured by the sample group, and D is the absorbance measured by the third control group.
II, experimental results:
the tyrosinase inhibition rates for each sample set were as follows:
single component N-acetylneuraminic acid:
| n-acetylneuraminic acid concentration (%) | Inhibition ratio (%) |
| 0.20％ | 32.8％ |
| 0.50％ | 57.7％ |
| 0.70％ | 85.9％ |
| 0.90％ | 93.1％ |
N-acetylneuraminic acid and nicotinamide (1: 1) group A1
| Composition concentration (%) | Inhibition ratio (%) |
| 0.20％ | 30.2％ |
| 0.50％ | 49.2％ |
| 0.70％ | 79.5％ |
| 0.90％ | 92.2％ |
N-acetylneuraminic acid and arbutin (1: 1) group A2
| Composition concentration (%) | Inhibition ratio (%) |
| 0.20％ | 48.9％ |
| 0.50％ | 60.3％ |
| 0.70％ | 83.5％ |
| 0.90％ | 95.2％ |
N-acetylneuraminic acid and vitamin C (1: 1) group A3
| Composition concentration (%) | Inhibition ratio (%) |
| 0.20％ | 36.3％ |
| 0.50％ | 45.5％ |
| 0.70％ | 79.6％ |
| 0.90％ | 90.3％ |
N-acetylneuraminic acid and cubilose extract (1: 1) group A4
| Composition concentration (%) | Inhibition ratio (%) |
| 0.20％ | 28.4％ |
| 0.50％ | 43.3％ |
| 0.70％ | 63.5％ |
| 0.90％ | 86.2％ |
N-acetylneuraminic acid, nicotinamide and arbutin (1: 1: 1) group A5
| Composition concentration (%) | Inhibition ratio (%) |
| 0.20％ | 35.2％ |
| 0.50％ | 48.7％ |
| 0.70％ | 81.2％ |
| 0.90％ | 94.0％ |
N-acetylneuraminic acid, nicotinamide and bird's nest extract (1: 1: 1) group A6
N-acetylneuraminic acid, nicotinamide, arbutin and cubilose extract (1: 1: 1: 1) group A7
| Composition concentration (%) | Inhibition ratio (%) |
| 0.20％ | 48.3％ |
| 0.50％ | 67.9％ |
| 0.70％ | 86.5％ |
| 0.90％ | 98.2％ |
And the detection of group a7 is shown in figure 1.
Under the same detection concentration, the whitening effect of the single-component N-acetylneuraminic acid under a certain concentration is better, and in the composition, the whitening effect of the A7 group under the same concentration is obviously better than that of other combined formulas. Whitening test of monocomponent N-acetylneuraminic acid and N-acetylneuraminic acid, nicotinamide, arbutin and cubilose extract group A7
First, experiment principle
The whitening effect of the test substance is evaluated by utilizing a keratinocyte and melanocyte co-culture system, and the test substance is considered to have the skin whitening effect if the test substance can reduce the generation of the total amount of melanin by co-culturing the keratinocyte and the melanocyte in a ratio simulating two cells in normal skin.
When the total melanin generation amount is detected, the melanin particles are dissolved in a high-temperature alkaline solution, and the generation amount of the melanin particles is indirectly reflected by utilizing the fact that the melanin particles have larger light absorption at 405 nm.
Second, Experimental methods
1. Cell culture and drug treatment
Prepared keratinocytes (Hacat cells) and melanocytes (B16 cells) were seeded at a ratio of 22:1 in 6-well cell plates (blank wells were set, only Hacat cells at the corresponding concentration were added to the blank wells). After 24 hours, 2ml of fresh PBS culture was added to each well of the negative control group, and 1ml of single-component N-acetylneuraminic acid of various concentrations tested and sample solution of group A7 (in PBS culture) was added to each well of the sample group. Monocomponent N-acetylneuraminic acid and group a7 were formulated at the following concentrations: 0.5% (0.5mg/ml), 0.7% (0.7mg/ml), 0.9% (0.9 mg/ml). 0.5mg/ml kojic acid was used as a positive control.
Each group is provided with 3 multiple holes for placing CO2Incubator (37 ℃, 5% CO)2) And (4) incubating. A second sample was added 48 hours later, 2ml of solution per well, as above. After another 48 hours of incubation, a third sample was added, 2ml of solution per well, as above. After the third sample addition for 24 hours, each group of cells was collected by digestion, centrifuged, and the supernatant was discarded.
2. Melanin content detection
To the collected cells, 200. mu.l of a melanin extract (1mol/L, NaOH + 10% DMSO) was added, and the mixture was uniformly pipetted and washed in a water bath at 95 ℃ for 1 hour. And after cooling, moderately centrifuging the tube wall liquid drops, uniformly blowing, sucking 150 mu l of solution in each centrifuge tube, transferring into a 96-well plate, and detecting the Abs405 value of each well by using an enzyme-labeling instrument.
The inhibition rate of melanin production was calculated:
in the formula: t is the sample well Abs; c is negative control well Abs; c0Blank wells Abs.
3. Determination of results
And judging whether the sample has the whitening effect according to the light absorption value measured at 405 nm.
In the melanin detection experiment, the OD value of the sample group is smaller than that of the negative control group, and the significant difference relative to the negative control group indicates that the sample has the effect of inhibiting the melanin generation.
In a cell tyrosinase activity detection experiment, the OD value of a sample group is smaller than that of a negative control group, and the significant difference of the OD value relative to the negative control group indicates that the sample has the effect of inhibiting tyrosinase.
Third, experimental results
1. The results of the assay of N-acetylneuraminic acid inhibition of melanin production by melanocytes are shown in the following table:
| experimental concentration (mg/ml) | 0.5 | 0.7 | 0.9 | Control group |
| Inhibition ratio (%) | 40 | 50 | 55 | 20 |
| Cell survival rate (%) | 100 | 100 | 100 | 100 |
The experimental result shows that the N-acetylneuraminic acid composition effectively inhibits the generation of melanin (compared with a control, the result is obviously different, P < 0.05.), and has the whitening effect.
2. The results of the tests of the N-acetylneuraminic acid, niacinamide, arbutin and bird's nest extract group A7 are shown in the following table:
| experimental concentration (mg/ml) | 0.5 | 0.7 | 0.9 | Control group |
| Inhibition ratio (%) | 45 | 53 | 67 | 20 |
| Cell survival rate (%) | 100 | 100 | 100 | 100 |
And (4) experimental conclusion: the N-acetylneuraminic acid has a good whitening effect, and can be used for realizing a synergistic effect with other whitening substances, so that the whitening effect of the N-acetylneuraminic acid composition is more remarkable. And the N-acetylneuraminic acid composition is a very safe whitening agent, and does not have any adverse effect on the normal growth of melanocytes under the effective dose.
Although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (7)
1. A whitening skin care product is characterized by comprising N-acetylneuraminic acid and a B-phase substance, wherein the B-phase substance is nicotinamide, arbutin and a cubilose extract; the ratio of the N-acetylneuraminic acid to the B-phase substance is 1:1 to 4.
2. The whitening skin care product of claim 1, further comprising a moisturizer and/or other acceptable adjuvants in the field of skin care products.
3. The whitening skin care product according to claim 2, wherein the ratio of the N-acetylneuraminic acid to the moisturizing agent is 0.1-3: 1.
4. The whitening skin care product of claim 2, wherein the moisturizer comprises one or more of butylene glycol, hexylene glycol, squalane, hyaluronic acid, panthenol, beta-glucan, and sodium polyglutamate.
5. The whitening skin care product of claim 4, wherein the humectant is a composition of squalane, hyaluronic acid, panthenol, beta-glucan and sodium polyglutamate, and the composition comprises 10-20% of squalane, 10-20% of hyaluronic acid, 20-40% of panthenol, 20-40% of beta-glucan and 20-40% of sodium polyglutamate.
6. The whitening skin care product according to claim 2, wherein the other acceptable adjuvants in the field of skin care products comprise one or more of solubilizing agents, preservatives, antioxidants, pH regulators, penetration enhancers, thickening agents, chelating agents, skin feel regulators, surfactants, emulsifiers, fragrances and pigments.
7. The whitening skin care product of claim 2, wherein the skin care product is in the form of a cream, emulsion, solution, film, aerosol or spray.
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| CN111743810A (en) * | 2020-06-29 | 2020-10-09 | 合肥中科华燕生物技术有限公司 | Moisturizing skin care product containing N-acetylneuraminic acid and preparation method thereof |
| CN112353807A (en) * | 2020-11-04 | 2021-02-12 | 嘉必优生物技术（武汉）股份有限公司 | Application of sialic acid in preparation of melanin pigmentation inhibitor |
| CN114344191A (en) * | 2021-12-27 | 2022-04-15 | 武汉中科光谷绿色生物技术有限公司 | Moisturizing mask liquid and mask |
| CN114668682B (en) * | 2022-04-29 | 2023-07-07 | 广州睿森生物科技有限公司 | Moisturizing and skin-brightening composition and application thereof |
| CN116370323A (en) * | 2023-02-21 | 2023-07-04 | 上海臻臣化妆品有限公司 | Whitening composition, skin care product and whitening liquid |
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| CN103025311B (en) * | 2010-05-25 | 2016-03-16 | 西姆莱斯有限公司 | As the carbamic acid menthyl ester compound of skin and/or hair whitening active thing |
| KR101386979B1 (en) * | 2010-10-20 | 2014-04-18 | 주식회사 베네비오 | Compositions for Hair Removal |
| CN104083291A (en) * | 2014-06-16 | 2014-10-08 | 武汉中科光谷绿色生物技术有限公司 | Application of N-acetylneuraminic acid monomer or its hydrate in cosmetics |
| CN108524416A (en) * | 2018-05-07 | 2018-09-14 | 安徽金龙山葛业有限公司 | A kind of skin care face cream containing pueraria lobata ingredient |
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