CN111686039B

CN111686039B - Whitening and skin-brightening composition and preparation method thereof

Info

Publication number: CN111686039B
Application number: CN202010637105.3A
Authority: CN
Inventors: 邓秀文; 徐华; 于忠国
Original assignee: Guangzhou Jichuang Yimei Biotechnology Co ltd
Current assignee: Guangzhou Jichuang Yimei Biotechnology Co ltd
Priority date: 2020-07-03
Filing date: 2020-07-03
Publication date: 2023-04-14
Anticipated expiration: 2040-07-03
Also published as: CN111686039A

Abstract

The invention provides a whitening and skin-brightening composition and a preparation method thereof. The whitening and skin-brightening composition comprises freeze-dried powder I, freeze-dried powder II and a solvent; the freeze-dried powder I comprises nicotinamide, the freeze-dried powder II comprises glutathione and ascorbic acid, and the solvent comprises tranexamic acid. The whitening and skin-brightening composition is characterized in that the effective components are prepared into freeze-dried powder I, freeze-dried powder II and a solvent which are matched for use, so that the effects of tyrosinase can be inhibited, a melanin generation pathway is cut off, the generation of melanin is reduced fundamentally, and the formed melanin can be dispersed; meanwhile, the whitening and skin-brightening composition can convert dopaquinone into pheomelanin, so that the conversion of eumelanin is reduced, free radicals can be eliminated, the normal oxygen delivery capability of blood is maintained, and the skin is enabled to be natural and glossy.

Description

Whitening and skin-brightening composition and preparation method thereof

Technical Field

The invention belongs to the field of cosmetic preparation, and relates to a whitening and skin-brightening composition and a preparation method thereof.

Background

With aging, the metabolism of human skin begins to slow down, the luster of the skin gradually fades, and melanin is continuously accumulated. Melanin is the most important pigment affecting the whitening of skin, and inhibiting the generation of melanin is the main purpose of whitening products. The commonly used whitening agent for inhibiting melanin generation mainly comprises AA2G, arbutin and derivatives thereof, kojic acid and derivatives thereof, vitamin C and derivatives thereof, endothelin antagonist, licoflavone, anthocyanin, and extracts of green tea, rhododendron, grape seed, rhodiola rosea and other plants.

The freeze-dried powder is sterile powder prepared by freezing liquid into a solid state in a sterile environment, and carrying out vacuum pumping to sublimate and dry water. The freeze-dried powder is prepared by adopting a vacuum freeze-drying method of a freeze dryer to freeze the water in the liquid medicine in advance, and then sublimating the frozen water in the liquid under a vacuum sterile environment, thereby obtaining freeze-dried powder. Generally speaking, the freeze-dried powder set comprises a bottle of liquid solvent and a bottle of powder, and the powder is dissolved in the solvent for use.

CN110227038A discloses a freeze-dried powder with whitening activity, a preparation method and an application thereof. The freeze-dried powder consists of tranexamic acid, nicotinamide, mannitol and ascorbyl glucoside, can effectively lighten melanin, can brighten skin color and whiten skin after long-term use, is not added with a chemical preservative, does not irritate the skin, is stable and does not change color, and is high in product activity, safe and effective. The freeze-dried powder can also be directly added into cosmetics of other liquid formulations, such as essence and moisturizing water for dissolving, or can be added into cosmetics of solid formulations after being mutually dissolved with a solvent, such as moisturizing cream for uniformly stirring for use, so as to enhance the effect.

However, the above lyophilized powder contains few effective whitening components, and the method for removing melanin is simple, so that the formation of melanin cannot be fundamentally cut off, and the formed melanin cannot be effectively targeted, and the whitening effect is not ideal.

Therefore, the development of a composition which is mild, non-irritant, and capable of permeating the effective components through the skin, reducing the generation of melanin, and dispersing the formed true melanin to achieve the effects of whitening and brightening the skin is a problem to be solved in the field.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to provide a whitening and skin-brightening composition and a preparation method thereof.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, the invention provides a whitening and skin-brightening composition, which comprises freeze-dried powder I, freeze-dried powder II and a solvent; the freeze-dried powder I comprises nicotinamide, the freeze-dried powder II comprises glutathione and ascorbic acid, and the solvent comprises tranexamic acid.

In the invention, the whitening and skin-brightening composition is divided into freeze-dried powder I, freeze-dried powder II and a solvent, wherein the freeze-dried powder I contains nicotinamide which can inhibit the synthesis of melanin; the ascorbic acid is combined with the glutathione in the freeze-dried powder II, so that the special raw material taste of the glutathione can be reduced, the property of the ascorbic acid which is easily denatured by illumination and oxygen is improved, the tranexamic acid in the solvent can help the effective components in the freeze-dried powder to permeate into the skin and promote absorption, and the freeze-dried powder I, the freeze-dried powder II and the solvent are matched with each other to achieve the effects of whitening and brightening the skin.

According to the invention, the main active ingredients are respectively prepared into the freeze-dried powder I and the freeze-dried powder II, so that on one hand, the storage is convenient, the skin whitening and brightening effects are prevented from being influenced by the denaturation caused by excessively complex formula ingredients in the storage process, and the irritation of the product is effectively reduced; on the other hand, the freeze-dried powder I and the freeze-dried powder II can be conveniently used by consumers, and the consumers can selectively choose the products according to the requirements of the consumers and the skin conditions, for example, the freeze-dried powder I or the freeze-dried powder II can be singly chosen to be mutually mixed with the solvent, or the two freeze-dried powders can be simultaneously chosen to be mutually mixed with the solvent.

On the contrary, if the freeze-dried powder is not prepared separately, the components of the freeze-dried powder are more complex, and the components are mutually influenced in the storage process to generate irritation to the skin of the human body; meanwhile, the two freeze-dried powder formulas contain the effective components for whitening and brightening the skin, so that the freeze-dried powder is used for normal skin, and the whitening effect is obvious. However, the epidermis layer of a user with sensitive skin is thin and sensitive, and a product with too strong efficacy cannot be used, so that the two types of freeze-dried powder are arranged, different using methods are designed, and the skin-sensitive skin-care product is suitable for any skin personnel to use.

As the preferable technical scheme of the invention, the volume ratio of the raw materials of the freeze-dried powder I, the raw materials of the freeze-dried powder II and the raw materials of the solvent is as follows: 1 (2.5-3.5), which can be, for example, 1.

As a preferable technical scheme of the invention, the freeze-dried powder I comprises 0.5-5% of nicotinamide, 3-8% of mannitol, 0.01-1% of nonapeptide-1, 0.05-0.5% of freeze-drying protective agent, 0.0005-0.01% of yeast extract, 0.1-1% of glycine, 0.1-0.3% of pH regulator and the balance of water, wherein the mass fraction of the raw materials of the freeze-dried powder I is 100%.

Wherein nonapeptide-1 and yeast extract can inhibit tyrosinase, cut off melanin generation pathway, and improve whitening effect by combining with nicotinamide. In the present invention, the mass fraction of nicotinamide may be 0.5 to 5%, for example, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, or 5%; the mass fraction of mannitol is 3 to 8%, and may be, for example, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, or the like; the mass fraction of the nonapeptide-1 is 0.01 to 1%, and may be, for example, 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, or 1%; the mass fraction of the lyoprotectant may be 0.05 to 0.5%, for example, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, or the like; the yeast extract may be present in an amount of 0.0005 to 0.01% by mass, for example, 0.0005%, 0.001%, 0.0015%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, or 0.01%; the glycine may be contained in an amount of 0.1 to 1% by mass, for example, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, or 1%; the mass fraction of the pH adjuster is 0.1 to 0.3%, and may be, for example, 0.1%, 0.12%, 0.15%, 0.18%, 0.2%, 0.22%, 0.25%, 0.28%, or 0.3%.

Preferably, the lyoprotectant includes trehalose.

Preferably, the pH adjuster comprises disodium hydrogen phosphate and/or sodium dihydrogen phosphate.

As a preferable technical scheme of the invention, the freeze-dried powder II comprises 2.5-7% of glutathione, 3-5% of ascorbic acid, 4-8% of mannitol and the balance of water by taking the mass fraction of the raw materials of the freeze-dried powder II as 100%.

The ascorbic acid has strong reducibility, is easy to be oxidized and denatured and turns yellow; glutathione is reduced glutathione, has antioxidant property, but has special odor. The combination of the ascorbic acid and the glutathione reduces the special raw material taste of the glutathione, improves the property of the ascorbic acid which is easy to be modified by illumination and oxygen, and has better product properties, whitening and antioxidant effects.

In the present invention, the mass fraction of glutathione may be 2.5 to 7%, for example, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, or the like; the ascorbic acid may be 3 to 5% by mass, for example, 3%, 3.2%, 3.5%, 3.8%, 4%, 4.2%, 4.5%, 4.6%, 4.8%, or 5%; the mannitol may be present in a mass fraction of 4 to 8%, for example, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, or 8%.

In a preferred embodiment of the present invention, the solvent comprises, based on 100% by mass of the raw materials of the solvent, 3 to 10% of tranexamic acid, 2 to 5% of propylene glycol, 0.3 to 1% of sodium hyaluronate, 0.05 to 0.2% of hydroxyethyl cellulose, and the balance of water.

Melanocytes producing melanin are present in the basal layer of the skin epidermis, and the vehicle can help the effective components penetrate into the skin to reach the basal layer for efficacy. In the present invention, the tranexamic acid may be present in a mass fraction of 3 to 10%, for example, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, or the like; the propylene glycol may be present in an amount of 2 to 5% by mass, for example, 2%, 2.2%, 2.5%, 2.8%, 3%, 3.5%, 4%, 4.2%, 4.8%, or 5%; the mass fraction of the sodium hyaluronate may be 0.3 to 1%, for example, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, or 1%; the mass fraction of the hydroxyethyl cellulose is 0.05 to 0.2%, and may be, for example, 0.05%, 0.08%, 0.1%, 0.12%, 0.15%, 0.18%, or 0.2%.

In a second aspect, the present invention also provides a method for preparing the whitening and skin-lightening composition according to the first aspect, the method comprising: and respectively preparing freeze-dried powder I, freeze-dried powder II and a solvent according to the formula amount to obtain the whitening and skin-brightening composition.

As a preferred technical scheme of the invention, the preparation method of the freeze-dried powder I comprises the following steps: mixing mannitol, nicotinamide, freeze-drying protective agent, glycine, pH regulator and water according to the formula, stirring, cooling, adding yeast extract and nonapeptide-1, filtering for sterilization, filling in a penicillin bottle, and freeze-drying to obtain freeze-dried powder I.

The temperature of the freeze-dried powder I during preparation is 45-55 ℃, and can be 45 ℃, 46 ℃, 47 ℃, 48 ℃, 49 ℃,50 ℃, 51 ℃, 52 ℃, 53 ℃, 54 ℃ or 55 ℃ and the like.

Preferably, the stirring speed is 300-500r/min, such as 300r/min, 320r/min, 350r/min, 380r/min, 400r/min, 420r/min, 450r/min, 480r/min or 500r/min.

Preferably, the temperature is reduced to 20-40 ℃, for example, 20 ℃, 22 ℃, 24 ℃, 25 ℃, 28 ℃,30 ℃, 32 ℃,35 ℃, 36 ℃, 38 ℃ or 40 ℃.

Preferably, the sterilization method is to perform sterilization by using a filter with a pore size of 0.22 μm.

As a preferable technical scheme of the invention, the preparation method of the freeze-dried powder II comprises the following steps: mixing mannitol, glutathione, ascorbic acid and water according to the formula ratio, stirring, filtering for sterilization, filling in a penicillin bottle, and freeze-drying to obtain the freeze-dried powder II.

The temperature of the freeze-dried powder II during preparation is 45-55 ℃, and can be, for example, 45 ℃, 46 ℃, 47 ℃, 48 ℃, 49 ℃,50 ℃, 51 ℃, 52 ℃, 53 ℃, 54 ℃ or 55 ℃ and the like.

As a preferable technical scheme of the invention, the preparation method of the solvent comprises the following steps: mixing sodium hyaluronate, tranexamic acid, hydroxyethyl cellulose, propylene glycol and water according to the formula ratio, stirring, sterilizing and filling in a penicillin bottle to obtain the solvent.

The solvent is prepared at a temperature of 80 to 90 ℃ during mixing, and may be, for example, 80 ℃, 81 ℃, 82 ℃, 83 ℃, 84 ℃, 85 ℃, 86 ℃, 87 ℃, 88 ℃, 89 ℃ or 90 ℃.

Preferably, the sterilization is performed in an autoclave.

As a preferable technical scheme of the invention, the preparation method comprises the following steps:

(1) Mixing mannitol, nicotinamide, a freeze-drying protective agent, glycine, a pH regulator and water according to the formula ratio at 45-55 ℃, stirring at 300-500r/min, cooling to 20-40 ℃, adding a yeast extract and nonapeptide-1, performing filtration sterilization by using a filter with the pore diameter of 0.22 mu m, filling into a penicillin bottle, and performing freeze drying to obtain freeze-dried powder I;

(2) Mixing mannitol, glutathione, ascorbic acid and water according to the formula ratio at 45-55 ℃, stirring at 300-500r/min, performing filtration sterilization by using a filter with the pore diameter of 0.22 mu m, filling into a penicillin bottle, and performing freeze drying to obtain freeze-dried powder II;

(3) Mixing sodium hyaluronate, tranexamic acid, hydroxyethyl cellulose, propylene glycol and water according to the formula ratio at the temperature of 80-90 ℃, stirring, sterilizing and filling to obtain the solvent;

the volume ratio of the raw materials of the freeze-dried powder I to the raw materials of the freeze-dried powder II to the raw materials of the solvent is 1 (0.8-1.2) to 2.5-3.5, and the whitening and skin brightening composition is obtained after the freeze-dried powder I, the freeze-dried powder II and the solvent are packaged.

The application method of the whitening and skin-brightening composition provided by the invention can be as follows: mixing solvent and lyophilized powder I, shaking gently until the lyophilized powder is completely dissolved, mixing the dissolved liquid and lyophilized powder II, shaking gently until the solution is completely dissolved, and applying the dissolved liquid on face.

The numerical ranges set forth herein include not only the points recited above, but also any points between the numerical ranges not recited above, and are not exhaustive of the particular points included in the ranges for reasons of brevity and clarity.

Compared with the prior art, the invention has the following beneficial effects:

(1) The effective components in the whitening and skin-lightening composition comprise nicotinamide, glutathione, ascorbic acid, yeast extract and nonapeptide-1, and the obtained composition can inhibit the action of tyrosinase, cut off a melanin generation pathway, radically reduce the generation of melanin, and disperse the formed melanin; meanwhile, the composition can convert dopaquinone into pheomelanin, so that the conversion of eumelanin is reduced, free radicals can be eliminated, the normal oxygen delivery capacity of blood is maintained, the skin circulation is accelerated, and the skin is lightened naturally, wherein after the whitening and skin-brightening composition is treated, the relative tyrosinase activity is 35.6-40.1%, the relative melanin content is 50.1-59.6%, and the melanin reduction amount of a subject is 21.9-25.5%;

(2) The whitening and skin-brightening composition provided by the invention is prepared by mixing and freeze-drying nicotinamide, yeast extract and nonapeptide-1 which are effective components, and then mixing and freeze-drying glutathione and ascorbic acid, on one hand, the ascorbic acid is combined with the glutathione, so that the special raw material taste of the glutathione is reduced, and the property of the ascorbic acid which is easily denatured by illumination and oxygen is also improved, so that the property is stable; on the other hand, the products obtained by separate combination can be matched according to the population without skin texture, and two kinds of freeze-dried powder are recommended to be matched with a solvent for the general population with normal skin texture; aiming at the people with sensitive skin, the application proportion of the freeze-dried powder is reasonably matched according to the requirements, and different application methods are designed, so that the freeze-dried powder is suitable for any skin-type person.

Detailed Description

The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitation of the present invention.

Example 1

The embodiment provides a whitening and skin-brightening composition, wherein raw materials for preparing lyophilized powder I, lyophilized powder II and a solvent respectively comprise the following components (wherein the percentages all represent mass fractions):

the preparation method comprises the following steps:

(1) Mixing mannitol, nicotinamide, trehalose, glycine, disodium hydrogen phosphate, sodium dihydrogen phosphate and water at 50 deg.C, stirring at 500r/min, cooling to below 40 deg.C, adding yeast extract and nonapeptide-1, filtering with 0.22 μm filter for sterilization, bottling in penicillin bottle, and freeze drying to obtain lyophilized powder I;

(2) Mixing mannitol, glutathione, ascorbic acid and water at 50 deg.C, stirring at 500r/min, filtering for sterilization, bottling in penicillin bottle, and freeze drying to obtain lyophilized powder II;

(3) Mixing sodium hyaluronate, tranexamic acid, hydroxyethyl cellulose, propylene glycol and water according to the formula amount at 80 ℃, stirring, sterilizing and filling to obtain the solvent;

the volume ratio of the freeze-dried powder I to the freeze-dried powder II to the solvent before freeze-drying is 1.

Example 2

The embodiment provides a whitening and skin-brightening composition, wherein raw materials for preparing freeze-dried powder I, freeze-dried powder II and a solvent respectively comprise the following components (wherein the percentages represent mass fractions):

the preparation method comprises the following steps:

(1) Mixing mannitol, nicotinamide, trehalose, glycine, disodium hydrogen phosphate, sodium dihydrogen phosphate and water at 45 deg.C, stirring at 300r/min, cooling to below 40 deg.C, adding yeast extract and nonapeptide-1, filtering with 0.22 μm filter for sterilization, bottling in penicillin bottle, and freeze drying to obtain lyophilized powder I;

(2) Mixing mannitol, glutathione, ascorbic acid and water in a formula amount at 55 ℃, stirring at 500r/min, filtering for sterilization, filling into a penicillin bottle, and freeze-drying to obtain lyophilized powder II;

Example 3

the preparation method comprises the following steps:

(1) Mixing mannitol, nicotinamide, trehalose, glycine, disodium hydrogen phosphate, sodium dihydrogen phosphate and water at 55 deg.C, stirring at 450r/min, cooling to below 40 deg.C, adding yeast extract and nonapeptide-1, filtering with 0.22 μm filter for sterilization, bottling in penicillin bottle, and freeze drying to obtain lyophilized powder I;

(2) Mixing mannitol, glutathione, ascorbic acid and water according to the formula ratio at 45 ℃, stirring at 350r/min, filtering for sterilization, filling in a penicillin bottle, and freeze-drying to obtain lyophilized powder II;

(3) Mixing sodium hyaluronate, tranexamic acid, hydroxyethyl cellulose, propylene glycol and water according to the formula amount at 85 ℃, stirring, sterilizing and filling to obtain the solvent;

wherein the volume ratio of the freeze-dried powder I to the freeze-dried powder II before freeze-drying to the solvent is 1.

Example 4

The difference from the example 1 is that the lyophilized powder I does not contain the nonapeptide-1, the mass fraction of the nicotinamide is increased to 5.5%, and the rest components and the mass fraction are the same as the example 1.

Example 5

The difference from the example 1 is that the freeze-dried powder I does not contain yeast extract, the mass fraction of nicotinamide is increased to 5.005%, and the rest components and the mass fraction are the same as the example 1.

Comparative example 1

The difference from example 1 is that the freeze-dried powder I does not contain nicotinamide, the mass fractions of nonapeptide-1 and glycine are respectively increased to 3% and 3%, and the rest components and mass fractions are the same as example 1.

Comparative example 2

The difference from the embodiment 1 is that the solvent does not contain tranexamic acid, the mass fraction of the propylene glycol is increased to 13 percent, and the rest components and the mass fraction are the same as the embodiment 1.

Comparative example 3

The difference from example 1 is that the raw materials for preparing the lyophilized powder II do not contain ascorbic acid, and the rest raw materials and preparation conditions are the same as those in example 1.

Comparative example 4

The difference from example 1 is that the raw materials for preparing the freeze-dried powder II do not contain ascorbic acid, the dosage of the glutathione is increased to 9%, and the rest raw materials and the preparation conditions are the same as those in example 1.

Comparative example 5

The comparative example provides a composition, which comprises freeze-dried powder and a solvent, and the preparation raw materials are as follows:

the preparation method comprises the following steps:

(1) Mixing mannitol, nicotinamide, trehalose, glycine, disodium hydrogen phosphate, sodium dihydrogen phosphate, mannitol, glutathione, ascorbic acid, tranexamic acid and water at 50 deg.C, stirring at 500r/min, cooling to below 40 deg.C, adding yeast extract and nonapeptide-1, filtering with 0.22 μm filter for sterilization, and freeze drying to obtain lyophilized powder;

(2) Mixing the sodium hyaluronate, the hydroxyethyl cellulose, the propylene glycol and the water according to the formula ratio at the temperature of 80 ℃, stirring, sterilizing and filling to obtain the solvent; wherein the volume ratio of the freeze-dried powder to the solvent before freeze-drying is 2.

Comparative example 6

the preparation method comprises the following steps:

(1) Mixing mannitol, nicotinamide, trehalose, glycine, disodium hydrogen phosphate, sodium dihydrogen phosphate, mannitol, glutathione, ascorbic acid and water at 50 deg.C, stirring at 500r/min, cooling to below 40 deg.C, adding yeast extract and nonapeptide-1, filtering with 0.22 μm filter for sterilization, and freeze drying to obtain lyophilized powder;

(2) Mixing sodium hyaluronate, tranexamic acid hydroxyethyl cellulose, propylene glycol and water according to the formula amount at 80 ℃, stirring, sterilizing and filling to obtain the solvent; wherein the volume ratio of the freeze-dried powder to the solvent before freeze-drying is 2.

Comparative example 7

The difference from example 1 is that nicotinamide and ascorbic acid are replaced in the preparation starting materials, namely: the preparation raw materials of the freeze-dried powder I do not contain nicotinamide, nicotinamide with equal mass is added into the freeze-dried powder II, meanwhile, the preparation raw materials of the freeze-dried powder II do not contain ascorbic acid, ascorbic acid with equal mass is added into the freeze-dried powder I, and the rest raw materials and the preparation conditions are the same as those in the embodiment 1.

Comparative example 8

The difference from example 1 is that nicotinamide and glutathione are substituted for each other in the preparation raw material, namely: the preparation raw materials of the freeze-dried powder I do not contain nicotinamide, the nicotinamide with equal mass is added into the freeze-dried powder II, meanwhile, the preparation raw materials of the freeze-dried powder II do not contain glutathione, the glutathione with equal mass is added into the freeze-dried powder I, and the rest raw materials and the preparation conditions are the same as those in the embodiment 1.

Performance test 1

The compositions provided in examples 1 to 5 and comparative examples 1 to 8 were subjected to a safety performance test by the following method:

(1) Haemolysis test of erythrocytes

Preparation of erythrocyte suspension: selecting healthy rabbit, taking 9mL of blood from heart, adding 1mL of 2% potassium oxalate solution, centrifuging, discarding supernatant, diluting the precipitate to 20mL with 20mmol/L PBS solution, and storing at 4 ℃ for later use. Selected samples were diluted to different concentrations with PBS solution and 5 concentration gradients were set for each sample. Adding 200 μ L of the above erythrocyte suspension (final concentration of the sample is controlled to be 5, 10, 20, 50, 100mg/mL respectively) into 10mL of diluent of the sample to be tested, taking distilled water as total blood-dissolving control, taking PBS solution as negative control, mixing gently, incubating at 37 deg.C for 30min, centrifuging at 2000r/min for 10min, collecting supernatant, and testing its absorbance at 560nm with spectrophotometer (A) ₅₆₀ ) Calculating the hemolysis rate according to the following formula;

a standard curve of hemolysis rate vs. sample concentration was plotted, and the sample concentration at which hemolysis occurred in 50% erythrocytes (HD) was calculated ₅₀ )。

(2) Protein denaturation experiments:

diluting the sample to 10g/L with PBS solution, collecting 10mL dilution of the sample to be tested, adding 200 μ L of the erythrocyte suspension, using distilled water as blank control, 1mg/mL Sodium Dodecyl Sulfate (SDS) solution as positive control, mixing gently, incubating at 37 deg.C for 30min, centrifuging at 2000r/min for 10min, collecting supernatant, and testing absorbance A at 540nm and 575nm with spectrophotometer ₅₄₀ And A ₅₇₅ Calculating a protein Denaturation Index (DI) according to the following formula;

wherein R is ₁ = blank control group a ₅₇₅ Blank control group A ₅₄₀ ，R ₂ = Experimental group A ₅₇₅ Experimental group A ₅₄₀ ，R ₃ = positive control group a ₅₇₅ Positive control group A ₅₄₀ 。

Evaluating the irritation of the sample to be tested according to the L/D value, wherein the L/D value is HD ₅₀ DI, erythrocyte hemolysis assay irritation grading criteria are shown in Table 1 below:

TABLE 1

L/D	Grading
		>100	Has no irritation
10<L/D≤100	Micro-stimulation property
		1<L/D≤10	Mild irritation
0.1<L/D≤1	Moderate irritation

The results of the above-described erythrocyte hemolysis test and protein denaturation test are shown in the following table 2:

TABLE 2

As can be seen from the safety performance test, the L/D values of the whitening and skin-brightening composition provided by the invention are all larger than 100, which indicates that the whitening composition is mild and has no stimulation.

Performance test 2

The compositions provided in examples 1 to 5 and comparative examples 1 to 8 were subjected to a whitening effect test by the following method:

(1) Tyrosinase activity inhibition assay

Mouse melanoma B16 cells in logarithmic growth phase were inoculated into 6-well cell culture plates and cultured overnight. The test samples with a final volume fraction of 1% were added to each of the test samples, and the untreated group was used as a cell control group, with 2 duplicate wells per group. After 48h of culture, the cells were washed 1 time with PBS, 100. Mu.L of lysate was added to each well, the cells were scraped off and collected, and the supernatant was centrifuged. 50 μ L of cell supernatant was applied to a 96-well plate, 50 μ L of 1-dopa solution was added, incubation was carried out at 37 ℃ for 1h, and absorbance was read at 475nm using an M3 plate reader.

Relative tyrosinase activity (%) = (assay well absorbance value-blank control absorbance value)/(cell control absorbance value-blank control absorbance value) × 100%.

(2) Experiment for inhibiting melanin synthesis

Mouse melanoma B16 cells in logarithmic growth phase were inoculated into T25 cell culture flasks and cultured overnight. The test samples with a final volume fraction of 1% were added to the untreated group as a cell control group. After 48h of culture, washing the cells with PBS for 1 time, adding 1mL of 1mol/L NaOH solution, scraping and collecting the cells, putting the cells into a water bath at 80 ℃ for 30min, adding the supernatant into a 96-well plate, and reading the absorbance value at 475nm by an M3 plate reader.

Relative melanin content (%) = (determination well absorbance value-blank control absorbance value)/(cell control absorbance value-blank control absorbance value) × 100%.

The results of the tyrosinase activity and melanin synthesis inhibition experiments are shown in table 3, wherein the tyrosinase activity of the control group is 100%, and the melanin content of the control group is 100%.

(3) Human body evaluation method for whitening effect

100 female volunteers between 25-50 years of age were selected and the compositions provided in examples 1-5 and comparative examples 1-8 were applied to the face 1 time each day in the morning and evening for a total of 4 weeks.

The evaluation method comprises the following steps: the change in melanin content of the skin before and after the application of the whitening composition to the subject was evaluated using a skin red melanin tester (Hexameter MX 18). The measurement range of the used instrument is 0-999, and the higher the measurement value, the higher the melanin content in the skin is.

The amount of melanin reduction in the skin of the back and front skin before the use of the whitening composition prepared according to the present invention is shown in table 3.

TABLE 3

The results in the table show that the whitening and skin-brightening composition provided by the invention can obviously inhibit the activity of tyrosinase and reduce the production of melanin, and can also obviously reduce the content of melanin on the face of a subject, wherein after the whitening and skin-brightening composition is treated, the activity of tyrosinase is 35.6-40.1%, the content of melanin is 50.1-59.6%, and the melanin reduction amount of the subject is 21.9-25.5%;

meanwhile, as can be seen from comparison between example 1 and comparative examples 1 to 4, the ingredients have a synergistic interaction, and if one of the ingredients is absent, the whitening effect is reduced;

as can be seen from comparison between example 1 and comparative examples 5 to 8, the whitening effect of example 1 is slightly superior to that of comparative examples 5 to 8, and therefore in order to further prove the advantages of the whitening and skin-lightening composition provided by the present invention, volunteer evaluation tests were performed in the present invention with respect to example 1 and comparative examples 5 to 8.

Performance test 3

Aiming at the people with sensitive skin, the whitening and skin-brightening composition provided by the invention can be reasonably matched with the use proportion of the freeze-dried powder according to the requirements, and different use methods are designed.

Selecting 100 female volunteers between 25-50 years of age, randomly dividing into 5 groups, and evaluating the compositions provided in example 1 and comparative examples 5-8; wherein the volunteers using example 1 were matched appropriately according to the individual skin type.

Wherein, the full score of the irritation score is 5, and the evaluation standard is as follows:

0.1-2 minutes, the upper face has a tingling sensation, and the skin is hot; 2.1-4 points, slight tingling sensation on the face, and skin fever; 4.1-4.5 minutes, no abnormal shape exists on the face, but the skin has slight fever; 4.5-5 points, no obvious abnormal shape on the face;

the full score of the experience feeling is 5, and the evaluation standard is as follows:

0.1-2 minutes, the upper face has sticky feeling, the texture is thick and heavy, the smearing is not easy to be even, and the skin problems such as acne or mouth closing and the like are easy to occur after the face-beautifying cream is used; 2.1 to 4 minutes, the face has sticky feeling, but the paste is easy to smear, and the skin problems such as slight mouth closing and the like occur after the paste is used; 4.1-4.5 points, fresh texture and no foreign body sensation on the face; 4.5-5 points, fresh and cool texture and comfortable face;

the score of the whitening effect is fully divided into 5 points, and the evaluation standard is as follows:

0.1-2 points, no obvious whitening effect but blackened; 2.1-4 minutes, without obvious whitening effect; 4.1-4.5 points, has whitening effect, but is not obvious; 4.5 to 5 minutes, and the color becomes white obviously after the use;

the scores of the data are shown in table 4:

TABLE 4

The above table shows that the whitening and skin-brightening composition provided by the invention can be reasonably matched with the use proportion of the freeze-dried powder according to requirements, so that the composition is friendly to the skin of a volunteer, is not easy to generate irritation, and has good experience, the evaluation of the volunteer on comparative examples 5-8 is obviously low, particularly the experience is one item, the composition is sticky to the volunteer, and the skin problems of slight mouth closing and the like occur after the composition is used.

In conclusion, the main active ingredients of the whitening and skin-brightening composition provided by the invention are respectively prepared into the freeze-dried powder I and the freeze-dried powder II, so that the storage is convenient, the whitening and skin-brightening effects are prevented from being influenced due to the denaturation caused by excessively complex formula ingredients in the storage process, the irritation of the product is effectively reduced, the use by consumers is facilitated, and the consumers can selectively select the product according to the requirements and skin conditions of the consumers through the formula ingredients of the freeze-dried powder I and the freeze-dried powder II, so that the whitening and skin-brightening effects are achieved.

The applicant declares that the above description is only a specific embodiment of the present invention, but the scope of the present invention is not limited thereto, and it should be understood by those skilled in the art that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention disclosed herein fall within the scope and disclosure of the present invention.

Claims

1. The whitening and skin-brightening composition is characterized by comprising freeze-dried powder I, freeze-dried powder II and a solvent;

the freeze-dried powder I comprises, by mass, 100% of raw materials of 0.5-5% of nicotinamide, 3-8% of mannitol, 0.01-1% of nonapeptide-1, 0.05-0.5% of a freeze-drying protective agent, 0.0005-0.01% of a yeast extract, 0.1-1% of glycine, 0.1-0.3% of a pH regulator and the balance of water;

the raw materials of the freeze-dried powder II are calculated by 100 percent in mass, and comprise 2.5 to 7 percent of glutathione, 3 to 5 percent of ascorbic acid, 4 to 8 percent of mannitol and the balance of water;

the solvent comprises, by mass, 100% of raw materials of tranexamic acid 3-10%, propylene glycol 2-5%, sodium hyaluronate 0.3-1%, hydroxyethyl cellulose 0.05-0.2%, and the balance of water.

2. The whitening and skin-brightening composition of claim 1, wherein the volume ratio of the raw materials of the lyophilized powder I, the lyophilized powder II and the solvent is 1 (0.8-1.2) to 2.5-3.5).

3. The whitening and skin-brightening composition of claim 2, wherein the volume ratio of the raw materials of the lyophilized powder I, the raw materials of the lyophilized powder II and the raw materials of the solvent is 1.

4. The whitening and skin-lightening composition of claim 1, wherein the lyoprotectant comprises trehalose.

5. The whitening and skin-lightening composition of claim 1, wherein the pH adjusting agent comprises disodium hydrogen phosphate and/or sodium dihydrogen phosphate.

6. The method of preparing the whitening and skin-lightening composition according to any one of claims 1 to 5, comprising:

and respectively preparing freeze-dried powder I, freeze-dried powder II and a solvent according to the formula amount to obtain the whitening and skin-brightening composition.

7. The preparation method of claim 6, wherein the preparation method of the lyophilized powder I comprises the following steps:

mixing mannitol, nicotinamide, freeze-drying protective agent, glycine, pH regulator and water according to the formula, stirring, cooling, adding yeast extract and nonapeptide-1, filtering for sterilization, filling in a penicillin bottle, and freeze-drying to obtain freeze-dried powder I.

8. The method according to claim 7, wherein the temperature during the mixing is 45 to 55 ℃.

9. The method of claim 7, wherein the stirring speed is 300 to 500r/min.

10. The method according to claim 7, wherein the temperature is reduced by adjusting the temperature to 20 to 40 ℃.

11. The method of claim 7, wherein the sterilization is performed by filtration using a filter having a pore size of 0.22 μm.

12. The preparation method of claim 6, wherein the preparation method of the lyophilized powder II comprises the following steps:

mixing mannitol, glutathione, ascorbic acid and water according to the formula ratio, stirring, filtering for sterilization, filling in a penicillin bottle, and freeze-drying to obtain freeze-dried powder II.

13. The method according to claim 12, wherein the temperature during the mixing is 45 to 55 ℃.

14. The method of claim 12, wherein the stirring speed is 300 to 500r/min.

15. The method of claim 12, wherein the sterilization is performed by filter sterilization using a filter having a pore size of 0.22 μm.

16. The method according to claim 6, wherein the solvent is prepared by the following steps:

mixing sodium hyaluronate, tranexamic acid, hydroxyethyl cellulose, propylene glycol and water according to the formula ratio, stirring, sterilizing and filling to obtain the solvent.

17. The method of claim 16, wherein the mixing temperature is 80 to 90 ℃.

18. The method of claim 16, wherein the stirring speed is 300 to 500r/min.

19. The method of claim 16, wherein the sterilization is performed in an autoclave.

20. The production method according to any one of claims 6 to 19, characterized by comprising the steps of: