CN117297992A

CN117297992A - Whitening composition and preparation method and application thereof

Info

Publication number: CN117297992A
Application number: CN202311032728.8A
Authority: CN
Inventors: 贺青; 刘露; 钟小超
Original assignee: Baocui Biotechnology Co ltd
Current assignee: Baocui Biotechnology Co ltd
Priority date: 2023-08-16
Filing date: 2023-08-16
Publication date: 2023-12-29
Anticipated expiration: 2043-08-16
Also published as: CN117297992B

Abstract

The invention provides a whitening composition and a preparation method thereof, and belongs to the technical field of cosmetics. The whitening composition comprises the following components in parts by weight: 10-20 parts of nicotinamide mononucleotide/dipropylene glycol solution, 1-10 parts of dew grass extract and 50-90 parts of water, wherein the nicotinamide mononucleotide/dipropylene glycol solution consists of 1 part of nicotinamide mononucleotide, 10-50 parts of dipropylene glycol and 6 parts of water. The nicotinamide mononucleotide, the dew grass extract and the dipropylene glycol are combined in a specific proportion, the synergistic effect is achieved, the whitening effect of the whitening composition is improved, and the problems of rough skin, darkness and the like caused by skin micro-inflammation are solved.

Description

Whitening composition and preparation method and application thereof

Technical Field

The invention relates to the field of cosmetics, in particular to a whitening composition and a preparation method and application thereof.

Background

People with colored or dark skin generally want to whiten the skin. The external environment is a major factor affecting whitening, such as ultraviolet rays, pressure, stay up, etc., wherein the contribution of ultraviolet rays is the largest. The anti-ultraviolet sun-screening agent has the advantages of sun screening, avoiding ultraviolet rays, being not ignored, and having the effect of the current living habit on melanin. Modern people have great living environment pressure, various road anger syndrome, unstable emotion, stay up, take away and the like, and the skin of people who are not eroded is avoided. The "brain-intestine-skin" axis is thought to affect the condition of the skin, especially because cortisol is always present in the blood of people under the above-mentioned survival pressure, and this substance causes weak inflammation in the skin, and the skin is in a micro-inflammatory condition for a long time, so that the skin is rough, dull, or even aging-accelerating. In vivo and in vitro studies of nicotinamide mononucleotide (nicotinamide mononucleotide, NMN) show that the nicotinamide mononucleotide has certain effects of resisting aging, protecting light, resisting oxidation, relieving and the like on skin while influencing the level of nicotinamide adenine dinucleotide (nicotinamide adenine dinucleotide, NAD+) to improve aging-related diseases. However, it is not known whether these studies are carried out in a mouse model or in vitro biochemical or cellular experiments, and the related skin care effects can be achieved only by oral administration/injection, and whether the effects are the same as in vivo ingestion on human skin. Furthermore, NMN is a water-soluble ingredient, which has poor permeability in the skin, affecting its role in cosmetics.

Disclosure of Invention

The purpose of the present disclosure is to overcome the shortcomings of the prior art, and provide a whitening composition, a preparation method and an application thereof, wherein the whitening composition can well solve the rough and dull skin caused by skin micro-inflammation, and has a whitening effect when being applied to cosmetics for a long time.

In order to achieve the above purpose, the technical scheme adopted by the present disclosure is as follows:

in a first aspect, a whitening composition is provided, comprising the following components in parts by weight: 10-20 parts of nicotinamide mononucleotide/dipropylene glycol solution, 1-10 parts of dew grass extract and 50-90 parts of water, wherein the nicotinamide mononucleotide/dipropylene glycol solution consists of 1 part of nicotinamide mononucleotide, 10-50 parts of dipropylene glycol and 6 parts of water.

The extract of herba Cynomorii is the product extracted from Cyanotis arachnoides C.B.Clarke, which is a Commelinaceae plant, and contains 20-hydroxyecdysterone (beta-ecdysone) as main ingredient, and has effects of promoting cell growth, stimulating dermis cell division, promoting protein synthesis, dispelling pathogenic wind, activating collaterals, promoting diuresis, relieving swelling, removing asthenic fever, dredging channels, relieving pain, etc., and can be used for treating rheumarthritis, limb numbness, etc., promoting production of skin peeling of silkworm, shrimp and crab, reducing cholesterol in human body, and lowering blood sugar concentration.

The invention combines nicotinamide mononucleotide, dipropylene glycol and a water weed extract to obtain a whitening composition, wherein the dipropylene glycol can promote the transdermal absorptivity of the nicotinamide mononucleotide, enhance the anti-inflammatory and repairing activities of the nicotinamide mononucleotide, and the water weed extract can enhance the metabolism and activation of cells, and the three are combined in a specific proportion to synergistically improve the whitening effect of the whitening composition and solve the problems of rough skin, darkness and the like caused by skin micro-inflammation.

In one embodiment, the dipropylene glycol is 30 to 35 parts by weight;

and/or, the dew grass extract is 5-6 parts by weight;

and/or, the nicotinamide mononucleotide/dipropylene glycol solution is 15-17 parts by weight.

The contents of dipropylene glycol, dew grass extract and nicotinamide mononucleotide/dipropylene glycol solution in the whitening composition affect the whitening effect of the whitening composition, and when the contents of dipropylene glycol, dew grass extract and nicotinamide mononucleotide/dipropylene glycol solution are in the above parts by weight, the resulting whitening composition is better.

In a second aspect, a method for preparing the whitening composition is provided, which comprises the following steps:

Preparation of nicotinamide mononucleotide/dipropylene glycol solution: uniformly mixing nicotinamide mononucleotide, water and dipropylene glycol, and performing high-voltage pulse electric field treatment for 5-15 times under the condition of 5-15kV/cm of pulse electric field intensity, 5-30us of pulse time and 0.5-2Hz of pulse frequency to obtain nicotinamide mononucleotide/dipropylene glycol solution;

preparing a whitening composition: uniformly mixing nicotinamide mononucleotide/dipropylene glycol solution, a dew grass extract and water, and performing high-voltage pulse electric field treatment for 3-9 times under the conditions of 6-16kV/cm pulse electric field intensity, 5-15us pulse time and 0.1-1.0Hz pulse frequency to obtain the whitening composition.

The high-voltage pulse electric field technology is used as an effective non-thermal processing technology, and has incomparable advantages in the aspects of processing time, energy loss, environmental pollution, energy transfer, food functional nutrition and flavor maintenance. Its role in food processing is mainly as follows: sterilizing, inactivating enzyme, extracting functional active substances and degrading residual pesticide.

The invention firstly uses a high-voltage pulse electric field technology to prepare the whitening composition, firstly mixes nicotinamide mononucleotide, water and dipropylene glycol to prepare a nicotinamide mononucleotide/dipropylene glycol solution by the high-voltage pulse electric field technology, then mixes the nicotinamide mononucleotide/dipropylene glycol solution, the dew grass extract and water to prepare the whitening composition, and under the action of the high-voltage pulse electric field, the nicotinamide mononucleotide and the dipropylene glycol form supermolecules interacted by hydrogen bonds and Van der Waals force under the condition of water, so that the transdermal absorbability of the nicotinamide mononucleotide is obviously improved, and the whitening effect of the whitening composition is improved.

In one embodiment, the conditions of the high voltage pulsed electric field treatment in the preparation of nicotinamide mononucleotide/dipropylene glycol solution are as follows: the pulse electric field strength is 8-10kV/cm, the pulse time is 18-20us, and the pulse frequency is 0.7-1Hz;

and/or, in the preparation of the whitening composition, the conditions of the high-voltage pulse electric field treatment are as follows: the pulse electric field intensity is 9-10kV/cm, the pulse time is 9-10us, and the pulse frequency is 0.5-0.6Hz.

In the process of preparing nicotinamide mononucleotide/dipropylene glycol solution and whitening composition, the treatment parameters of the high-voltage pulsed electric field are critical, and when the treatment conditions of the high-voltage pulsed electric field are within the above preferred ranges, the obtained whitening composition has better whitening effect.

In a third aspect, there is provided the use of the whitening composition in the preparation of a cosmetic.

In a fourth aspect, there is provided a cosmetic comprising the above-described whitening composition.

In one embodiment, the whitening composition has a mass of 1-10% of the cosmetic mass.

In the present invention, the mass percentage of the whitening composition in the cosmetic may be 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5%, 10.0%.

The whitening composition with a specific proportion is added into the cosmetics, so that good whitening effect can be brought to the cosmetics. When the cosmetic of the present application contains less than 2% (w/w) of the above-described whitening composition of the present application, a sufficient whitening effect to enhance skin cannot be expected; and when it is contained in an amount exceeding 10% (w/w), an undesired reaction such as allergy or the like may occur or there is a problem in skin safety.

The compositions of the present invention may be incorporated into all types of carriers. Non-limiting examples include emulsions (e.g., water-in-oil-in-water, silicone-in-water, water-in-silicone, oil-in-water-in-oil, oil-in-silicone-in-water emulsions), creams, lotions, solutions (aqueous or water-ethanol solutions), anhydrous vehicles (e.g., lipsticks and powders), gels, and ointments. Variants and other suitable vectors will be apparent to any of ordinary skill in the art and are suitable for use in the present invention. In certain aspects, the concentrations and combinations of compounds, ingredients, and reagents are selected in the following manner: in such a way that the combination is chemically compatible and does not form complexes that precipitate out of the final product.

The cosmetic product of the present invention may also comprise any number of combinations of additional ingredients described throughout this specification (e.g., pigments or additional cosmetic or pharmaceutical ingredients). In the cosmetic, the concentration of the additional ingredient may vary. For example, in non-limiting embodiments, the composition can comprise, consist essentially of, or consist of, in its final form: for example, at least about 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.0010%, 0.5000%, 1.0%, 5.0%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99% or any derivable range therebetween. In a non-limiting aspect, the percentages can be calculated by weight or volume of the entire composition. It will be appreciated by those of ordinary skill in the art that the concentration in a given cosmetic product may vary depending on the addition, substitution, and/or reduction of ingredients.

In addition to the combinations of ingredients disclosed herein, the cosmetic may also contain additional ingredients, which are cosmetic ingredients, non-limiting examples of which are described in the subsection below.

CTFA international cosmetic ingredient dictionary and handbook (2004 and 2008) describe a wide variety of non-limiting cosmetic ingredients that can be used in the context of the present invention. Examples of such component species include: fragrances (artificial and natural), dyes and coloring components (e.g., titanium dioxide, D & C blue No. 4, D & C green No. 5, D & C orange No. 4, D & C red No. 33, D & C violet No. 2, D & C yellow No. 10), adsorbents, lubricants, solvents, humectants (including, for example, emollients, humectants, film formers, occlusive agents and agents that affect the natural moisturizing mechanism of the skin), water repellents, ultraviolet absorbers (physical and chemical absorbers, such as para-aminobenzoic acid ("PABA") and corresponding PABA derivatives, titanium dioxide, zinc oxide, and the like), essential oils, vitamins (e.g., A, B, C, D, E and K), trace metals (e.g., zinc, calcium, and selenium), anti-irritants (e.g., steroids and non-steroidal anti-inflammatory drugs), plant extracts (e.g., aloe vera, citrus, cucumber extract, ginkgo biloba, ginseng, and rosemary), antimicrobial agents, antioxidants (e.g., BHT and tocopherol), chelating agents (e.g., disodium edetate and tetrasodium edetate), preservatives (e.g., methylparaben and propylparaben), pH adjusting agents (e.g., sodium hydroxide and citric acid), absorbents (e.g., aluminum starch octenyl succinate, kaolin, corn starch, oat starch, cyclodextrin, talc, and zeolite), skin bleaching and brightening agents (e.g., hydroquinone and nicotinamide lactate), humectants (e.g., sorbitol, urea, methylglucitol polyether-20, and mannitol), exfoliants, preservatives, and, waterproofing agents (e.g., sodium hydroxide magnesium/aluminum stearate), skin conditioning agents (e.g., aloe vera extract, allantoin, bisabolol, ceramides, polydimethylsiloxane, sodium hyaluronate, bioglycol-1, ethylhexyl glycerol, pentanediol, hydrogenated polydecene, octyldodecanol oleate, and dipotassium glycyrrhizinate). Several non-limiting examples of these additional ingredients are provided in the following subsections.

UV absorbers that may be used in combination with the compositions of the present invention include chemical and physical sunscreens. Non-limiting examples of chemical sunscreens that may be used include para-aminobenzoic acid (PABA), PABA esters, isoamyl p-methoxycinnamate, octocrylene, octyl triazone, terephthalylidene dicarbanone sulfonic acid. Non-limiting examples of physical sunscreens include kaolin, talc, petrolatum and metal oxides (e.g., titanium dioxide and zinc oxide).

Non-limiting examples of humectants that can be used with the compositions of the present invention include amino acids, chondroitin sulfate, glycerin, butylene glycol, propylene glycol, xylitol, erythritol, fructose, rhamnose, oleic acid, olive (olea europaea) oil, chondroitin sodium sulfate, sodium hyaluronate, sodium lactate, sodium palmitate, sodium PCA, sodium polyglutamate, soluble collagen, tocopherol, tocopheryl acetate, tocopherol, and mixtures thereof.

Non-limiting examples of antioxidants that may be used with the compositions of the present invention include acetylcysteine, ascorbyl polypeptide, ascorbyl dipalmitate, ascorbyl methylsilanol pectate, ascorbyl palmitate, ascorbyl stearate, BHA, BHT, t-butylhydroquinone, cysteine HCI, dipentylhydroquinone, di-t-butylhydroquinone, dicetyl thiodipropionate, thiosalicylic acid, tocopheryl polyoxyethylene-5 ether, tocopheryl polyoxyethylene-10 ether, tocopheryl polyoxyethylene-12 ether, tocopheryl polyoxyethylene-18 ether, and mixtures thereof.

Non-limiting examples of emulsifiers that may be used with the compositions of the present invention include esters of glycerin, esters of propylene glycol, fatty acid esters of polyethylene glycol, fatty acid esters of polypropylene glycol, esters of sorbitol dehydrate, carboxylic acid copolymers, esters and ethers of glucose, ethoxylated ethers, ethoxylated alcohols, alkyl phosphates, polyoxyethylene fatty ether phosphates, PPG-5 cetyl polyether-20, bis-PEG/PPG-20/20 dimethoxysilane, cetyl polyoxyethylene-10 ether, polysorbate-80, and mixtures thereof.

Thickeners may also increase the stability of the cosmetic product of the present invention. Non-limiting examples of additional thickeners that may be used include carboxylic acid polymers, crosslinked polyacrylate polymers, polyacrylamide polymers, polysaccharides, gums, hydrogenated polyisobutene, glycerol tristearate, ammonium acryloyldimethyl taurate/VP copolymer, or mixtures thereof.

Non-limiting examples of preservatives that may be used include quaternary ammonium salt preservatives (e.g., polyquaternium-1 and benzalkonium halides (e.g., benzalkonium chloride ("BAC") and benzalkonium bromide)), parabens (e.g., methyl and propyl parabens), phenoxyethanol, benzyl alcohol, chlorobutanol, phenol, sorbic acid, or combinations thereof.

In one embodiment, the cosmetic product is a cream comprising the following components in weight percent: 3-5% of glycerol, 2-8% of butanediol, 0.1-0.5% of cellulose, 0.05-0.2% of sodium hyaluronate, 0.1-1% of acrylic acid (esters)/C10-30 alkanol acrylate cross-linked polymer, 2-8% of butter fruit tree fruit fat, 2-8% of caprylic acid/capric acid triglyceride, 3-6% of polydimethylsiloxane and 5-10% of glycerol stearate; 2-10% of a whitening composition; 0.1-1% of 1,2 hexanediol, 0.1-1% of p-hydroxyacetophenone and the balance of water.

In one embodiment, the cosmetic product is a facial mask comprising the following components in percentage by weight: 3-5% of glycerin, 0.1-0.3% of xanthan gum, 0.05-0.2% of sodium hyaluronate, 1-5% of whitening composition, 0.1-1% of 1,2 hexanediol, 0.1-1% of p-hydroxyacetophenone and the balance of water.

Compared with the prior art, the beneficial effects of the present disclosure are: the invention combines nicotinamide mononucleotide, dipropylene glycol and the dew grass extract to obtain the whitening composition, wherein the dipropylene glycol can promote the transdermal absorptivity of the nicotinamide mononucleotide, enhance the activity of the nicotinamide mononucleotide, and the dew grass extract can enhance the metabolism and activation of cells, and the three are combined in a specific proportion to synergistically improve the whitening effect of the whitening composition, so that the problems of rough skin, darkness and the like caused by skin micro-inflammation are solved.

Detailed Description

For a better understanding of the objects, technical solutions and advantages of the present disclosure, the present disclosure will be further described with reference to specific examples and comparative examples, which are intended to be in detail, not to be limiting of the present disclosure. All other embodiments, which can be made by one of ordinary skill in the art without making any inventive effort, are intended to be within the scope of the present disclosure. The experimental reagents and apparatus to which the present disclosure is directed are common reagents and apparatus unless otherwise indicated.

In the following examples and comparative examples, unless otherwise specified, the parts are parts by weight; nicotinamide mononucleotide is abbreviated as NMN, nicotinamide mononucleotide aqueous solution is abbreviated as NMN aqueous solution, and nicotinamide mononucleotide/dipropylene glycol solution is abbreviated as NMN/dipropylene glycol solution; the extract is prepared from herba Avenae Fatuae powder below 40 mesh.

Example 1

The invention relates to a whitening composition, a whitening mask and a whitening cream.

The preparation method of the whitening composition comprises the following steps:

preparing an aqueous NMN solution: adding 1 part of NMN into 6 parts of deionized water, and dissolving at normal temperature to obtain an NMN aqueous solution;

Preparing NMN/dipropylene glycol solution: adding NMN aqueous solution into 30 parts of dipropylene glycol, uniformly mixing, and introducing the obtained mixed solution into a high-voltage pulser, and performing high-voltage pulsed electric field treatment for 9 times under the conditions of the pulsed electric field strength of 10kV/cm, the pulse time of 18us and the pulse frequency of 1Hz to obtain NMN/dipropylene glycol solution;

preparing a whitening composition: and uniformly mixing 15 parts of NMN/dipropylene glycol solution, 5 parts of dew grass extract and 75 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument to perform high-voltage pulse electric field treatment for 6 times under the conditions of 9kV/cm of pulse electric field intensity, 10us of pulse time and 0.5Hz of pulse frequency to obtain the whitening composition.

The preparation method of the mask comprises the following steps:

uniformly mixing 4 parts of glycerin, 0.2 part of xanthan gum, 0.1 part of sodium hyaluronate and 80 parts of deionized water, heating the obtained mixed solution to 85 ℃, preserving heat for 30min, stirring and cooling to 60 ℃, adding 0.5 part of p-hydroxyacetophenone and 0.5 part of 1, 2-hexanediol, stirring and dissolving, cooling to 45 ℃, adding 3 parts of the whitening composition of the embodiment, uniformly stirring, adding 100 parts of deionized water, and uniformly stirring to obtain a mask solution;

and adding 25mL of mask liquid into the mask bag filled with the mask cloth, and enabling the mask cloth to fully contact the mask liquid to obtain the whitening mask.

The composition and preparation method of the face cream are as follows:

phase A: 4 parts of glycerin, 5 parts of butanediol, 0.2 part of cellulose, 0.1 part of sodium hyaluronate, 0.5 part of acrylic acid (ester)/C10-30 alkanol acrylate cross-linked polymer and 60 parts of deionized water;

and B phase: 6 parts of butter tree fruit fat, 5.5 parts of caprylic/capric triglyceride, 4.5 parts of polydimethylsiloxane and 6.7 parts of glycerol stearate;

and C phase: 6 parts of a whitening composition;

and D phase: 0.5 part of 1,2 hexanediol, 0.5 part of p-hydroxyacetophenone and 5 parts of deionized water;

weighing phase A according to a proportion, uniformly mixing, heating to 85 ℃, and preserving heat for 30min for standby;

weighing phase B according to a proportion, uniformly mixing, heating to 85 ℃, and preserving heat for 30min for standby;

weighing the phase D according to a proportion, and uniformly mixing for later use;

pouring the phase B into the phase A at a speed of 5g/min at a homogenizing speed of 800r/min, homogenizing for 5min at a speed of 1500r/min, stirring and cooling to 60 ℃, adding the phase D, stirring uniformly, cooling to 45 ℃, adding the phase C, adding deionized water to 100 parts, and stirring uniformly to obtain the whitening cream.

Example 2

The preparation method of the whitening composition of this example is different from that of the whitening composition of example 1 only in that: in the preparation of NMN/dipropylene glycol solution, 10 parts of dipropylene glycol was used.

The whitening mask was prepared according to the preparation method of the whitening mask of example 1 using the whitening composition of this example.

The whitening cream was prepared according to the preparation method of the whitening cream of example 1 using the whitening composition of this example.

Example 3

The preparation method of the whitening composition of this example is different from that of the whitening composition of example 1 only in that: in the preparation of NMN/dipropylene glycol solution, 50 parts of dipropylene glycol was used.

Example 4

The preparation method of the whitening composition of this example is different from that of the whitening composition of example 1 only in that: in the preparation of NMN/dipropylene glycol solution, 35 parts of dipropylene glycol was used.

Example 5

The preparation method of the whitening composition of this example is different from that of the whitening composition of example 1 only in that: different steps are used to prepare NMN/dipropylene glycol solutions;

the procedure for preparing the NMN/dipropylene glycol solution in this example was: adding NMN aqueous solution into 30 parts of dipropylene glycol, uniformly mixing, introducing the obtained mixed solution into a high-voltage pulser, and carrying out high-voltage pulsed electric field treatment for 5 times under the conditions of 5kV/cm pulsed electric field intensity, 30us pulsed time and 2Hz pulsed frequency to obtain NMN/dipropylene glycol solution.

Example 6

the procedure for preparing the NMN/dipropylene glycol solution in this example was: adding NMN aqueous solution into 30 parts of dipropylene glycol, uniformly mixing, introducing the obtained mixed solution into a high-voltage pulser, and performing high-voltage pulsed electric field treatment for 15 times under the conditions of pulsed electric field intensity of 15kV/cm, pulse time of 5us and pulse frequency of 0.5Hz to obtain NMN/dipropylene glycol solution.

Example 7

the procedure for preparing the NMN/dipropylene glycol solution in this example was: adding NMN aqueous solution into 30 parts of dipropylene glycol, uniformly mixing, introducing the obtained mixed solution into a high-voltage pulser, and performing high-voltage pulsed electric field treatment for 12 times under the conditions of 8kV/cm of pulsed electric field intensity, 20us of pulsed time and 0.7Hz of pulsed frequency to obtain NMN/dipropylene glycol solution.

Example 8

The preparation method of the whitening composition of this example is different from that of the whitening composition of example 1 only in that: the steps for preparing the whitening composition are different;

the steps for preparing the whitening composition in this example are: uniformly mixing 10 parts of NMN/dipropylene glycol solution, 1 part of dew grass extract and 50 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 6 times under the conditions of 9kV/cm of pulse electric field intensity, 10us of pulse time and 0.5Hz of pulse frequency to obtain the whitening composition.

Example 9

the steps for preparing the whitening composition in this example are: and uniformly mixing 20 parts of NMN/dipropylene glycol solution, 10 parts of dew grass extract and 90 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 6 times under the conditions of 9kV/cm of pulse electric field intensity, 10us of pulse time and 0.5Hz of pulse frequency to obtain the whitening composition.

Example 10

the steps for preparing the whitening composition in this example are: and (3) uniformly mixing 17 parts of NMN/dipropylene glycol solution, 6 parts of dew grass extract and 80 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 6 times under the conditions of 9kV/cm of pulse electric field intensity, 10us of pulse time and 0.5Hz of pulse frequency to obtain the whitening composition.

Example 11

the steps for preparing the whitening composition in this example are: and uniformly mixing 15 parts of NMN/dipropylene glycol solution, 5 parts of dew grass extract and 75 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 9 times under the conditions of 6kV/cm of pulse electric field intensity, 15us of pulse time and 0.1Hz of pulse frequency to obtain the whitening composition.

Example 12

the steps for preparing the whitening composition in this example are: and (3) uniformly mixing 15 parts of NMN/dipropylene glycol solution, 5 parts of dew grass extract and 75 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 3 times under the conditions of 6kV/cm of pulse electric field intensity, 15us of pulse time and 1.0Hz of pulse frequency to obtain the whitening composition.

Example 13

the steps for preparing the whitening composition in this example are: and (3) uniformly mixing 15 parts of NMN/dipropylene glycol solution, 5 parts of dew grass extract and 75 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 3 times under the conditions of pulse electric field intensity of 16kV/cm, pulse time of 5us and pulse frequency of 1.0Hz to obtain the whitening composition.

Example 14

the steps for preparing the whitening composition in this example are: and uniformly mixing 15 parts of NMN/dipropylene glycol solution, 5 parts of dew grass extract and 75 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 9 times under the conditions of pulse electric field intensity of 16kV/cm, pulse time of 5us and pulse frequency of 0.1Hz to obtain the whitening composition.

Example 15

the steps for preparing the whitening composition in this example are: and (3) uniformly mixing 15 parts of NMN/dipropylene glycol solution, 5 parts of dew grass extract and 75 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 5 times under the conditions of 10kV/cm of pulse electric field intensity, 9us of pulse time and 0.6Hz of pulse frequency to obtain the whitening composition.

Example 16

The invention relates to a whitening mask and an embodiment of whitening cream.

The whitening mask of this embodiment differs from the whitening mask of embodiment 1 only in that: the whitening composition is 1 part.

The whitening cream of this embodiment differs from the whitening cream of embodiment 1 only in that: the whitening composition is 2 parts.

Example 17

The invention relates to a whitening mask and an embodiment of whitening cream.

The whitening mask of this embodiment differs from the whitening mask of embodiment 1 only in that: the whitening composition is 5 parts.

The whitening cream of this embodiment differs from the whitening cream of embodiment 1 only in that: the whitening composition is 10 parts.

Example 18

The invention relates to a whitening mask and an embodiment of whitening cream.

The whitening mask of this embodiment differs from the whitening mask of embodiment 1 only in that: the whitening composition is 2 parts.

The whitening cream of this embodiment differs from the whitening cream of embodiment 1 only in that: the whitening composition is 7 parts.

Comparative example 1

A comparative example of the whitening composition according to the present invention, the preparation method of the whitening composition according to the present comparative example is different from that of the whitening composition according to example 1 only in that: different steps for preparing the NMN aqueous solution;

The procedure for preparing the aqueous NMN solution in this comparative example was: 1 part of NMN is added into 5 parts of deionized water, and the mixture is dissolved at normal temperature to obtain an NMN aqueous solution.

Comparative example 2

the procedure for preparing the aqueous NMN solution in this comparative example was: 1 part of NMN is added into 7 parts of deionized water, and the mixture is dissolved at normal temperature to obtain an NMN aqueous solution.

Comparative example 3

A comparative example of the whitening composition according to the present invention, the preparation method of the whitening composition according to the present comparative example is different from that of the whitening composition according to example 1 only in that: in the preparation of NMN/dipropylene glycol solution, 8 parts of dipropylene glycol was used.

Comparative example 4

A comparative example of the whitening composition according to the present invention, the preparation method of the whitening composition according to the present comparative example is different from that of the whitening composition according to example 1 only in that: in the NMN/dipropylene glycol solution, 52 parts of dipropylene glycol was prepared.

Comparative example 5

A comparative example of the whitening composition according to the present invention, the preparation method of the whitening composition according to the present comparative example is different from that of the whitening composition according to example 1 only in that: different steps are used to prepare NMN/dipropylene glycol solutions;

The procedure for preparing NMN/dipropylene glycol solution in this comparative example was: adding NMN aqueous solution into 30 parts of dipropylene glycol, uniformly mixing, introducing the obtained mixed solution into a high-voltage pulser, and performing high-voltage pulsed electric field treatment for 4 times under the conditions of 4kV/cm of pulsed electric field intensity, 32us of pulsed time and 2.5Hz of pulsed frequency to obtain NMN/dipropylene glycol solution.

Comparative example 6

the procedure for preparing NMN/dipropylene glycol solution in this comparative example was: adding NMN aqueous solution into 30 parts of dipropylene glycol, uniformly mixing, introducing the obtained mixed solution into a high-voltage pulser, and carrying out high-voltage pulsed electric field treatment for 17 times under the conditions of 17kV/cm of pulsed electric field intensity, 4us of pulsed time and 0.4Hz of pulsed frequency to obtain NMN/dipropylene glycol solution.

Comparative example 7

The procedure for preparing NMN/dipropylene glycol solution in this comparative example was: adding NMN aqueous solution into 30 parts of dipropylene glycol, uniformly mixing, introducing the obtained mixed solution into a high-voltage pulser, and performing high-voltage pulsed electric field treatment for 17 times under the conditions of 4kV/cm of pulsed electric field intensity, 4us of pulsed time and 2.5Hz of pulsed frequency to obtain NMN/dipropylene glycol solution.

Comparative example 8

the procedure for preparing NMN/dipropylene glycol solution in this comparative example was: adding NMN aqueous solution into 30 parts of dipropylene glycol, uniformly mixing, introducing the obtained mixed solution into a high-voltage pulser, and performing high-voltage pulsed electric field treatment for 4 times under the conditions of 17kV/cm of pulsed electric field intensity, 32us of pulsed time and 2.5Hz of pulsed frequency to obtain NMN/dipropylene glycol solution.

Comparative example 9

A comparative example of the whitening composition according to the present invention, the preparation method of the whitening composition according to the present comparative example is different from that of the whitening composition according to example 1 only in that: the steps for preparing the whitening composition are different;

the procedure for preparing the whitening composition in this comparative example is: and uniformly mixing 9 parts of NMN/dipropylene glycol solution, 0.8 part of dew grass extract and 48 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 6 times under the conditions of 9kV/cm of pulse electric field intensity, 10us of pulse time and 0.5Hz of pulse frequency to obtain the whitening composition.

Comparative example 10

the procedure for preparing the whitening composition in this comparative example is: and (3) uniformly mixing 22 parts of NMN/dipropylene glycol solution, 12 parts of dew grass extract and 93 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 6 times under the conditions of 9kV/cm of pulse electric field intensity, 10us of pulse time and 0.5Hz of pulse frequency to obtain the whitening composition.

Comparative example 11

the procedure for preparing the whitening composition in this comparative example is: and (3) uniformly mixing 9 parts of NMN/dipropylene glycol solution, 0.8 part of dew grass extract and 93 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 6 times under the conditions of 9kV/cm of pulse electric field intensity, 10us of pulse time and 0.5Hz of pulse frequency to obtain the whitening composition.

Comparative example 12

the procedure for preparing the whitening composition in this comparative example is: and (3) uniformly mixing 22 parts of NMN/dipropylene glycol solution, 12 parts of dew grass extract and 48 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 6 times under the conditions of 9kV/cm of pulse electric field intensity, 10us of pulse time and 0.5Hz of pulse frequency to obtain the whitening composition.

Comparative example 13

the procedure for preparing the whitening composition in this comparative example is: uniformly mixing 15 parts of NMN/dipropylene glycol solution, 5 parts of dew grass extract and 75 parts of deionized water, and magnetically stirring the obtained mixed solution at 60 ℃ for 3 hours to obtain the whitening composition.

Comparative example 14

The procedure for preparing the whitening composition in this comparative example is: and uniformly mixing 15 parts of NMN/dipropylene glycol solution, 5 parts of dew grass extract and 75 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 10 times under the conditions of 5kV/cm of pulse electric field intensity, 16us of pulse time and 0.09Hz of pulse frequency to obtain the whitening composition.

Comparative example 15

the procedure for preparing the whitening composition in this comparative example is: and uniformly mixing 15 parts of NMN/dipropylene glycol solution, 5 parts of dew grass extract and 75 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 2 times under the conditions of 5kV/cm of pulse electric field intensity, 16us of pulse time and 1.1Hz of pulse frequency to obtain the whitening composition.

Comparative example 16

The procedure for preparing the whitening composition in this comparative example is: and uniformly mixing 15 parts of NMN/dipropylene glycol solution, 5 parts of dew grass extract and 75 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 10 times under the conditions of 17kV/cm of pulse electric field intensity, 4us of pulse time and 1.1Hz of pulse frequency to obtain the whitening composition.

Comparative example 17

the procedure for preparing the whitening composition in this comparative example is: and uniformly mixing 15 parts of NMN/dipropylene glycol solution, 5 parts of dew grass extract and 75 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 2 times under the conditions of 17kV/cm of pulse electric field intensity, 4us of pulse time and 0.09Hz of pulse frequency to obtain the whitening composition.

Comparative example 18

A comparative example of the whitening composition of the present invention, the preparation method of the whitening composition of the present comparative example is as follows:

preparation of dipropylene glycol solution: adding 7 parts of deionized water into 30 parts of dipropylene glycol, uniformly mixing, and introducing the obtained mixed solution into a high-voltage pulser, and carrying out high-voltage pulsed electric field treatment for 9 times under the condition of the pulsed electric field strength of 10kV/cm and the pulse time of 18us and the pulse frequency of 1Hz to obtain a dipropylene glycol solution;

Preparing a whitening composition: and uniformly mixing 15 parts of dipropylene glycol solution, 5 parts of the dew grass extract and 75 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 6 times under the conditions of 9kV/cm of pulse electric field intensity, 10us of pulse time and 0.5Hz of pulse frequency to obtain the whitening composition.

Comparative example 19

preparing a whitening composition: and uniformly mixing 15 parts of NMN aqueous solution, 5 parts of the dew grass extract and 75 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulse instrument, and performing high-voltage pulse electric field treatment for 6 times under the conditions of 9kV/cm of pulse electric field intensity, 10us of pulse time and 0.5Hz of pulse frequency to obtain the whitening composition.

Comparative example 20

Preparing a whitening composition: and (3) uniformly mixing 15 parts of NMN/dipropylene glycol solution and 80 parts of deionized water, and introducing the obtained mixed solution into a high-voltage pulser, and performing high-voltage pulsed electric field treatment for 6 times under the conditions of 9kV/cm of pulsed electric field intensity, 10us of pulse time and 0.5Hz of pulse frequency to obtain the whitening composition.

Comparative example 21

A comparative example of the whitening composition according to the present invention, the preparation method of the whitening composition according to the present comparative example is different from that of the whitening composition according to example 1 only in that: in preparing the whitening composition, 5 parts of rose powder is used instead of 5 parts

Dew grass extract.

Comparative example 22

A comparative example of the whitening composition according to the present invention, the preparation method of the whitening composition according to the present comparative example is different from that of the whitening composition according to example 1 only in that: in preparing the whitening composition, 5 parts of lotus powder is used for replacing 5 parts of lotus powder

Dew grass extract.

Comparative example 23

A comparative example of the whitening composition according to the present invention, the preparation method of the whitening composition according to the present comparative example is different from that of the whitening composition according to example 1 only in that: in preparing the whitening composition, 5 parts of the extract of the dew grass was replaced with 5 parts of ascorbyl glucoside.

Comparative example 24

A comparative example of the whitening composition according to the present invention, the preparation method of the whitening composition according to the present comparative example is different from that of the whitening composition according to example 1 only in that: in preparing the NMN/dipropylene glycol solution, 30 parts of dipropylene glycol was replaced with 30 parts of 1, 2-propanediol.

Comparative example 25

A comparative example of the whitening composition according to the present invention, the preparation method of the whitening composition according to the present comparative example is different from that of the whitening composition according to example 1 only in that: in preparing the NMN/dipropylene glycol solution, 30 parts of dipropylene glycol was replaced with 30 parts of 1, 3-propanediol.

Comparative example 26

A comparative example of the whitening composition according to the present invention, the preparation method of the whitening composition according to the present comparative example is different from that of the whitening composition according to example 1 only in that: in preparing the NMN/dipropylene glycol solution, 30 parts of dipropylene glycol was replaced with 30 parts of glycerol.

Comparative example 27

A comparative example of the whitening composition according to the present invention, the preparation method of the whitening composition according to the present comparative example is different from that of the whitening composition according to example 1 only in that: in preparing the NMN/dipropylene glycol solution, 30 parts of dipropylene glycol was replaced with 30 parts of 1, 3-butanediol.

Comparative example 28

A comparative example of the whitening composition according to the present invention, the preparation method of the whitening composition according to the present comparative example is different from that of the whitening composition according to example 1 only in that: in preparing the NMN/dipropylene glycol solution, 30 parts of dipropylene glycol was replaced with 30 parts of polyglycerol-10.

Comparative example 29

A comparative example of the whitening mask and the whitening cream of the invention.

The whitening mask of this comparative example differs from the whitening mask of example 1 only in that: the whitening composition is 0.8 part.

The whitening cream of this comparative example differs from the whitening cream of example 1 only in that: the whitening composition is 1 part.

Comparative example 30

The whitening mask of this comparative example differs from the whitening mask of example 1 only in that: the whitening composition is 6 parts.

The whitening cream of this comparative example differs from the whitening cream of example 1 only in that: the whitening composition is 12 parts.

Comparative example 31

Comparative example 32

Effect example 1

This effect example tests the performance of the NMN/dipropylene glycol solution obtained in example 1, the whitening compositions obtained in examples 1 to 15 and comparative examples 1 to 28 as follows:

1. NMN transdermal absorption rate: respectively adding equal volumes of the whitening compositions of examples 1-15 and comparative examples 1-28 into a container, taking fresh treated pigskin, cutting into a proper size according to the caliber of a diffusion tank, and fully contacting the dermis layer with a receiving liquid after the surface (cuticle layer) of the pigskin faces upwards after the detection and no damage; removing bubbles, contacting the bright white factor with stratum corneum, setting the temperature to 33 ℃, and taking out 0.5mL of receiving solution (PBS) at 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12h respectively, wherein the volume of the receiving solution (PBS) is 4.5 mL; and simultaneously supplementing an equal volume of PBS and removing bubbles, detecting the NMN content according to an HPLC method, and calculating the accumulated transdermal rate.

2. Inflammation of the bodyInhibition rate of disease factor: according to the test group setup, 0.9mL of epidermal cell TA culture solution was added to a 6-well plate, and then a 3D epidermal model was appliedTransferring to a 6-hole plate, and marking test group numbers on the 6-hole plate; wherein, the epidermal cell TA culture solution and the 3D epidermal model +.>All purchased from Shaanxi Boxi biotechnology Co. Taking PBS as solvent of sample group (examples 1-15 and comparative examples 1-28), adding prepared working solution 25uL with different concentrations on the surface of 3D epidermis model, gently shaking the 3D epidermis model to uniformly distribute the samples on the surface of the 3D epidermis model, placing in CO ₂ Incubator (37 ℃ C., volume fraction 5% CO) ₂ 95% relative humidity) for 24h. After the incubation, the test substance remained on the surface of the 3D epidermis model is washed by a washing bottle filled with sterile PBS solution, and the inner and outer residual liquid of the 3D epidermis model is gently wiped by a sterile cotton swab.

(1) Tissue viability assay the washed 3D epidermis model was placed in 24 well plates containing 1mg/mL MTT working solution. Transfer of the 24-well plate to CO ₂ In incubator (37 ℃, volume fraction 5% CO) ₂ 95% relative humidity) for 3h. After the MTT incubation was completed, the 3D skin model was removed with forceps, the MTT liquid remaining on the bottom surface was wiped with absorbent paper, transferred to a new 24-well plate, 2mL of isopropyl alcohol was added, the 24-well plate was sealed with a sealing film, and left to stand at 4℃overnight. After leaching, the 3D skin model was pierced with a 200 pipette tip and isopropyl alcohol leaching solution was flowed from the 3D skin model into a 24-well plate. The pierced 3D epidermis model was discarded, and the isopropanol extract in each well was blown 3 times and thoroughly mixed. After mixing, 2 parts of 200uL of isopropanol extract was extracted from each well and added to the corresponding wells of the 96-well plate, respectively, and labeled. The absorbance value is read by a microplate reader at 570nm wavelength. Based on the safe concentration of MTT, a blank group (PBS solution), examples 1 to 15, and comparative examples 1 to 28 were set to perform an inflammatory factor inhibition experiment at the same experimental concentration.

(2) After the incubation for detecting inflammatory factors is finished, collecting model culture solution in an EP tube, and storing in a refrigerator at-80 ℃. Detection analysis was performed according to the instructions of the Elisa detection kit for IL-1. Alpha. Inflammatory factors. IL-1 a inhibition = (blank-sample) blank/100%.

3. Tyrosinase inhibition rate: with minor modifications to the reference T/SHRH 015-2018, "cosmetic-tyrosinase inhibition assay method", a sample (whitening compositions of examples 1-15 and comparative examples 1-28) solution was added to a 10mL volumetric flask, diluted to 0.01, 0.05, 0.20, 0.50, 0.80mg/mL with PBS at pH 6.8, and loaded: respectively sucking 0.5mL of sample solution and 0.5mL of 100 mu g/mL of tyrosinase solution into a 5mL centrifuge tube, uniformly mixing, then carrying out water bath at 37 ℃ for 10min, adding 2mL of 1.0mg/mL of levodopa solution into the centrifuge tube, standing for 5min, immediately measuring the absorbance at 475nm, and marking as Ai; the tyrosinase solution was replaced with 0.5mL of PBS, and the absorbance was measured and designated Aj; the sample solution was replaced with 0.5mL of PBS, and the absorbance was measured and recorded as Bi; 1mL of PBS was pipetted in place of the sample and tyrosine solution and the absorbance was measured and recorded as Bj. Tyrosinase inhibition rate/% = (1- (Ai-Aj)/(Bi-Bj)) @ 100%.

4. MITF (microphthalmia-associated transcription factor) expression inhibition rate:

1) Cell culture human A375 melanoma cells were cultured in DMEM medium containing 10% fetal bovine serum, 100U/mL penicillin and 100. Mu.g/mL streptomycin at 37℃with 5% CO ₂ Culturing in incubator, and changing 1 time of culture medium every 3 d.

2) Western blot detection of protein expression safe dose concentrations of the sample groups (examples 1-15 and comparative examples 1-28) were determined using MTT, and the same experimental concentrations were set within safe concentration ranges. The vehicle control and the same concentration (examples 1-15 and comparative examples 1-28) dosing groups were set with 3 replicates per concentration. Cells were collected 48h after administration, total cellular proteins were extracted, and protein concentration was measured. Protein loading was 50. Mu.g, and after electrophoresis, transfer and blocking, MITF antibody (1:1000) was added and incubated overnight at 4 ℃. After buffer elution, the mixture was incubated in horseradish peroxidase-labeled goat anti-rabbit IgG (1:2000) at room temperature for 2h, and ECL reaction was performed with MITF antibody as an internal reference. The ratio of the absorbance of the MITF fragment bands was compared with the relative expression level. MITF expression inhibition = (sample group-blank group)/sample group x 100%.

5. Beta-ecdysone content: beta-ecdysone is used as a standard substance, beta-ecdysone standard solutions of 0.005, 0.01, 0.02, 0.04 and 0.08mg/mL are accurately prepared, the content of beta-ecdysone is calculated according to the peak area in a chromatogram, the experiment is repeated 3 times, the average value is calculated, the peak area in the chromatogram is taken as an ordinate y, a standard curve is drawn by taking the mass concentration of a sample to be detected as an abscissa x, and the content of ecdysone is calculated according to the standard curve. Detecting by high performance liquid chromatography, and purifying by chromatographic column: XDB-C18 (4.6X105 mm,5 um); mobile phase: methanol: water = 40:60; flow rate: 0.8mL/min; detection wavelength: 246nm; column temperature: 31 ℃; sample injection amount: 10 mu L.

The test results are shown in Table 1.

TABLE 1

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As shown in the test results of Table 1, the transdermal absorption rate, tyrosinase inhibition rate and MITF expression inhibition rate of the whitening composition obtained by the present invention are all more than 50%.

As can be seen from a comparison of example 1 and comparative examples 1-2, NMN and deionized water were present in a mass ratio of 1:6 can form NMN saturated water solution in proportion, if NMN and deionized water form unsaturated or supersaturated water solution, the transdermal absorption rate, tyrosinase inhibition rate and MITF expression inhibition rate of the obtained whitening composition are obviously reduced.

Comparative examples 1 to 4 and comparative examples 3 to 4 show that the content of dipropylene glycol affects the performance of the whitening composition, and when the content of dipropylene glycol is out of the range of the present invention, the transdermal absorption rate, tyrosinase inhibition rate and MITF expression inhibition rate of the obtained whitening composition are significantly reduced.

Comparative example 1, examples 5 to 7 and comparative examples 5 to 8 show that when the parameters for preparing the NMN/dipropylene glycol solution deviate from the protective scope of the present invention, the transdermal absorption rate, tyrosinase inhibition rate and MITF expression inhibition rate of the obtained whitening composition are significantly reduced.

As is apparent from comparative examples 1, 8-10 and 9-12, the contents of the respective components in the whitening composition affect the performance of the whitening composition, and when the dew grass extract is 5-6 parts by weight and the nicotinamide mononucleotide/dipropylene glycol solution is 15-17 parts by weight, the transdermal absorption rate, tyrosinase inhibition rate and MITF expression inhibition rate of the obtained whitening composition are higher.

Comparative examples 1 and 13 show that the whitening composition treated with the high-voltage pulse electric field has higher transdermal absorption rate, tyrosinase inhibition rate and MITF expression inhibition rate than the whitening composition prepared by conventional heating.

Comparative example 1, examples 11 to 15 and comparative examples 14 to 17 revealed that when the high-voltage pulsed electric field treatment parameters for preparing the whitening composition deviate from the scope of the present invention, the transdermal absorption rate, tyrosinase inhibition rate and MITF expression inhibition rate of the resulting whitening composition are significantly reduced.

Comparative example 1 and comparative examples 18 to 20 show that the three components of the present invention synergistically enhance the whitening effect of the whitening composition.

Comparative example 1 and comparative examples 21 to 28 revealed that there was no synergy between the other components, except for the synergy between dipropylene glycol, the dew grass extract and NMN.

The data of comparative examples 1,8-15 and comparative examples 9-17 show that the best levels of beta-ecdysone are obtained only at the ratios of the present protective NMN/dipropylene glycol solution, the dew grass extract; under the condition of the pulse electric field, the beta-ecdysone can be relatively more dissolved in the whitening composition, so that the whitening composition has good whitening effect.

Effect example 2

This effect example tests the masks and creams prepared in examples 1, 16-18 and comparative examples 29-32; wherein, the whitening mask of example 1 was used alone for test group 1; the whitening cream of example 1 was used alone in test group 2.

Whitening facial mask and whitening facial cream.

The whitening efficacy testing method comprises the following steps:

selection of subjects: and selecting qualified subjects according to the inclusion and exclusion criteria, and ensuring that the number of the effective cases finally completed in each test area is not less than 30.

1.1 criteria for enrollment

1.1.1 volunteers aged 25-35 years, women with sun protection habits;

1.1.2 the skin color ITA degree value of the test part is 20-41 degrees;

1.1.3 no allergic diseases, no allergic history of cosmetics or other external preparations;

1.1.4 past history of no light susceptibility disease, no drugs affecting light susceptibility are used in recent times;

1.1.5 skin at the test site should be free of pigmentation, inflammation, scar, mole, and hair;

1.1.6 can receive a person whose skin is tanned in the test area using an artificial light source;

1.1.7 can understand the testing procedure, voluntarily attend the test and sign written informed consent.

1.2 exclusion criteria

1.2.1 pregnant or lactating women, or those having a pregnancy-preparing plan in the near future;

1.2.2 patients with skin history such as psoriasis, eczema, atopic dermatitis, severe acne, etc.;

1.2.3 oral or topical anti-inflammatory agents such as corticosteroids in approximately 1 month;

1.2.4 oral or topical administration over a period of approximately 2 months with any product or drug that affects skin color (e.g., hydroquinone-based formulations);

1.2.5 a similar test was performed within approximately 3 months or a similar test was performed 3 months ago, but the skin blackened marks at the test sites did not completely fade;

1.2.6 other clinical trials were enrolled in approximately 2 months;

3.1.2.7 other clinical evaluations were considered unsuitable for the participating test subjects.

2 test article

2.1 test product: masks and creams prepared in examples 1, 16-18 and comparative examples 29-32.

2.2 negative control: blackened area blank.

2.3 positive control: 7% ascorbic acid (vitamin C) product (refrigerated at 4deg.C, aluminum tube preserved in dark place) formulated according to whitening efficacy test appendix I formulation of cosmetic safety Specification 2015.

2.4 application of test substance

The staff smears the test object according to the random table corresponding to the test area, the sample smearing area should be not less than 6cm ² The coating amount is 2.00+/-0.05 mg/cm ² . The spacing between each test zone should be no less than 1.0cm. The product frequency should be according to the product instruction, if need to smear many times a day, each time smear interval is not less than 4 hours.

3 test site: the back is preferably selected as the test site, and non-exposure sites such as the thigh, upper arm, etc. may be selected. Each blackening test area should be not less than 0.5cm ² And should be located within each of the application areas.

4 test instrument

4.1 sunlight simulator: a xenon arc lamp solar simulator capable of generating ultraviolet rays of uva+uvb wavelengths with continuous spectral radiation is used. Wavelengths below 290nm are removed using a suitable filter system and the output spectrum is subjected to metrology or calibration.

4.2 skin colorimeter: there is an instrument that can measure L x a x b x color space data formulated by the international commission on illumination (CIE).

4.3 skin melanin detector: there is an instrument for detecting skin MI values based on the principle of spectral absorption.

5 environmental conditions

In the test process, the visual evaluation and the instrument test link are carried out under the environment that the temperature is 21+/-1 ℃ and the relative humidity is 50+/-10% RH, the visual evaluation is carried out under the condition of constant illumination (fluorescent tube or LED illumination with the color temperature of 5500-6500K), and the test subject can carry out the evaluation and the test after adapting for at least 30 minutes under the environment condition.

6 test procedure

6.1 enrolled in groups of volunteer subjects on demand, signed written informed consent. Before the group is put into the group, the subjects are inquired about a series of problems about disease history, health condition and the like according to the inclusion and exclusion criteria and the like, and meanwhile, the skin of the tested part is subjected to compliance evaluation and skin color test screening and recorded.

And 6.2, enabling the qualified subjects to enter a stage of establishing a human skin blackening model. The ultraviolet biological dose (MED) at the test site of each subject should first be determined. Then, each test area was selected at the test site, and irradiated 1 time per day with 0.75 times MED at the same irradiation point by a solar simulator for 4 days.

6.3 skin darkening period was 4 days after the end of irradiation, without any treatment.

6.4 on day 5 after the irradiation, visual evaluation and skin color instrument detection are carried out on the skin color of each test area, and the test area with poor consistency (the area with ITA degree value differing from the average value of all the test areas by more than 5) is removed. The corresponding test subjects were smeared according to a random table at each blackening test zone beginning the day.

6.5 continuous application of the test substance for at least 4 weeks, visual assessment and instrumental detection of skin color at 1 week, 2 weeks, 3 weeks and 4 weeks after application, and recording.

6.5.1 visual assessment

The skin color of each test area is evaluated by a dermatologist by means of a color chart from light to dark, and the scores are recorded in time. Comparing the skin color changes before and after the experiment, and calculating and scoring the skin color changes before and after the experiment according to a ten-step system (the more obvious the change is, the larger the score is);

consumers self-score according to the change condition of the skin color before and after the test, and self-score with ten scores (the more obvious the change is, the larger the score is).

6.5.2 skin colorimeter measurement

At each visit time point, the L, a and b values of each test area are measured by a skin colorimeter respectively, each area is tested three times, the ITA value is recorded and calculated, the larger the ITA value is, the lighter the skin color is, and otherwise, the darker the skin color is.

6.5.3 measurement by skin melanin detector

At each visit point, respectively measuring the MI value of each test area by using a skin melanin detector, testing each test area for three times, and recording; the smaller the MI value, the lower the skin melanin content, and conversely the higher the skin melanin content.

7 data analysis

Statistical analysis of the data was performed using statistical analysis software. The metering data are expressed as: the mean value is +/-standard deviation, normal distribution inspection is carried out, normal distribution requirements are met, paired t inspection is adopted for comparison before and after the mean value is self, and otherwise, two related sample rank sum inspection is adopted; comparing the grade data before and after use, and adopting two related sample ranks and tests; the comparison between the test area and the control area adopts independent sample t test or rank sum test; meanwhile, the regression coefficient (slope k value) of each parameter with time is calculated, and the significance level is P < 0.05.

The test results are shown in Table 2.

TABLE 2

As is clear from examples 1, 16 to 18 and comparative examples 29 to 32, the whitening composition added within the scope of the present invention has a good whitening effect. The addition amount of the whitening composition is insufficient, and the whitening effect is poor; the excessive addition amount of the whitening composition may cause heavy skin burden and poor final effect, and the whitening effect is only optimal when the whitening composition is used in the scope of the invention;

As can be seen from the comparison of example 1 and test groups 1-2, the use of the whitening mask together with the whitening cream can significantly increase the whitening effect of the cosmetic.

Finally, it should be noted that the above embodiments illustrate the technical solution of the present disclosure and not limit the scope of the present disclosure, and although the present disclosure has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the present disclosure may be modified or equally substituted without departing from the spirit and scope of the technical solution of the present disclosure.

Claims

1. The whitening composition is characterized by comprising the following components in parts by weight: 10-20 parts of nicotinamide mononucleotide/dipropylene glycol solution, 1-10 parts of dew grass extract and 50-90 parts of water, wherein the nicotinamide mononucleotide/dipropylene glycol solution consists of 1 part of nicotinamide mononucleotide, 10-50 parts of dipropylene glycol and 6 parts of water.

2. The whitening composition according to claim 1, wherein the dipropylene glycol is 30 to 35 parts by weight;

and/or, the dew grass extract is 5-6 parts by weight;

3. A method for preparing the whitening composition according to any one of claims 1 to 2, comprising the steps of:

4. The method of claim 3, wherein the conditions of the high-voltage pulsed electric field treatment in the nicotinamide mononucleotide/dipropylene glycol solution are as follows: the pulse electric field strength is 8-10kV/cm, the pulse time is 18-20us, and the pulse frequency is 0.7-1Hz;

5. Use of a whitening composition according to any of claims 1-2 for the preparation of cosmetics.

6. A cosmetic product, characterized in that it contains the whitening composition according to any one of claims 1 to 2.

7. The cosmetic according to claim 6, wherein the whitening composition has a mass of 1 to 10% of the mass of the cosmetic.

8. The cosmetic according to claim 6 or 7, wherein the cosmetic is at least one of a lotion, an emulsion, a cream, a mask, a serum, and a spray.

9. The cosmetic product of claim 8, wherein the cosmetic product is a face cream, wherein the face cream comprises the following components in percentage by weight: 3-5% of glycerol, 2-8% of butanediol, 0.1-0.5% of cellulose, 0.05-0.2% of sodium hyaluronate, 0.1-1% of acrylic acid (esters)/C10-30 alkanol acrylate cross-linked polymer, 2-8% of butter fruit tree fruit fat, 2-8% of caprylic acid/capric acid triglyceride, 3-6% of polydimethylsiloxane and 5-10% of glycerol stearate; 2-10% of a whitening composition; 0.1-1% of 1,2 hexanediol, 0.1-1% of p-hydroxyacetophenone and the balance of water.

10. The cosmetic product of claim 8, wherein the cosmetic product is a facial mask comprising the following components in weight percent: 3-5% of glycerin, 0.1-0.3% of xanthan gum, 0.05-0.2% of sodium hyaluronate, 1-5% of whitening composition, 0.1-1% of 1,2 hexanediol, 0.1-1% of p-hydroxyacetophenone and the balance of water.