A skin whitening, rejuvenation and brightening composition and its preparation method and application

Abstract

The present invention provides a skin whitening and brightening composition and a preparation method and application thereof, and belongs to the technical field of skin care products. The skin whitening and brightening composition of the present invention comprises: niacinamide, phenethyl resorcinol nanoparticles, arbutin, lilac oil, pearl extract, purslane extract, allantoin and trehalose. The niacinamide, phenethyl resorcinol nanoparticles, arbutin and pearl extract of the present invention have the effects of reducing pigmentation, brightening skin tone and improving dullness; lilac oil can enhance the penetration of whitening ingredients into the deep layer of the skin, thereby achieving a whitening effect in a short time; purslane extract and allantoin have the effects of soothing the skin; trehalose can enhance the stress resistance of cells and reduce the damage of free radicals to the skin. The experimental results show that the various components of the composition of the present invention have synergistic effects, are mild and non-irritating to the skin, can reduce the generation and deposition of melanin in a relatively short time, and thus achieve the effect of whitening, brightening and brightening the skin.

Description

Technical Field

[0001] The invention relates to the technical field of skin care products, and in particular to a skin whitening, rejuvenation and brightening composition and a preparation method and application thereof. Background Art

[0002] In recent years, among all skin care needs, the demand for whitening has increased rapidly.

[0003] In terms of ingredients, among the current top whitening ingredients, the effects of popular ingredients such as fruit acid / salicylic acid, VC, and niacinamide are highly recognized and have a high degree of consumer awareness. However, there is always a game between concentration and safety of these ingredients.

[0004] There are many whitening products on the market that rarely meet the two major requirements of "mild + efficient". Therefore, it is urgent to invent a mild and efficient whitening product. Summary of the invention

[0005] The purpose of the present invention is to provide a skin whitening, rejuvenation and brightening composition and a preparation method and application thereof. The composition of the present invention is mild and non-irritating, can improve and reduce the generation and deposition of melanin in a relatively short period of time, and make the skin more lustrous, bright, smooth and delicate.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The invention provides a skin whitening, rejuvenating and brightening composition, comprising the following components in parts by weight: 10-20 parts of niacinamide, 10-20 parts of phenethylresorcinol nanoparticles, 8-12 parts of arbutin, 5-10 parts of lilac flower oil, 5-10 parts of pearl extract, 10-20 parts of purslane extract, 3-8 parts of allantoin and 3-8 parts of trehalose.

[0008] Preferably, the preparation method of the phenethylresorcinol nanoparticles comprises: dissolving phospholipids in methanol, adding a cyclodextrin aqueous solution and stirring, removing methanol to obtain an aqueous phase; dissolving phenethylresorcinol in anhydrous ethanol, adding the aqueous phase, removing ethanol, and freeze-drying to obtain the phenethylresorcinol nanoparticles.

[0009] More preferably, the phospholipids include one or two of soybean lecithin, hydrogenated soybean lecithin or egg yolk lecithin; and the cyclodextrin is water-soluble cyclodextrin.

[0010] More preferably, the mass ratio of the phospholipid, cyclodextrin and phenethylresorcinol is 1:2-4:1-3.

[0011] Preferably, the preparation method of the pearl extract comprises: crushing the pearls and mixing them with glucose, inoculating composite microorganisms and fermenting for 24-36 hours, filtering to obtain a fermentation liquid, adding neutral protease for enzymatic hydrolysis, filtering under ultra-high pressure, and freeze-drying to obtain the pearl extract.

[0012] More preferably, the composite microorganism is Bacillus terequina and Fusarium venezuelae.

[0013] Preferably, the preparation method of the purslane extract comprises: drying and crushing the purslane, performing ethanol microwave extraction, filtering and freeze-drying to obtain the purslane extract.

[0014] The present invention also provides a method for preparing the above-mentioned skin whitening, rejuvenating and brightening composition, comprising: mixing and stirring the auxiliary materials with the niacinamide, phenylethylresorcinol nanoparticles, arbutin, lilac flower oil, pearl extract, purslane extract, allantoin and trehalose to obtain the composition.

[0015] Preferably, the auxiliary materials include emulsifiers, emollients, humectants, thickeners, preservatives, flavors and deionized water.

[0016] The present invention also provides application of the skin whitening, rejuvenation and brightening composition in the preparation of cosmetics.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] The present invention provides a skin whitening and brightening composition, comprising: niacinamide, phenethyl resorcinol nanoparticles, arbutin, lilac oil, pearl extract, purslane extract, allantoin and trehalose. Among them, niacinamide, phenethyl resorcinol nanoparticles, arbutin and pearl extract have the effects of inhibiting the transfer of melanin to epidermal cells, reducing pigmentation, brightening skin color and improving dullness; lilac oil can enhance the penetration of whitening ingredients into the deep layer of the skin, so as to achieve whitening effect in a short time; purslane extract has the effect of soothing the skin. Allantoin has a soothing and calming effect, making the skin smoother. Trehalose can enhance the stress resistance of cells, reduce the damage of free radicals to the skin, and protect skin cells from damage. Experimental results show that the various components of the composition of the present invention have synergistic effects, the composition is mild and non-irritating to the skin, can improve and reduce the generation and deposition of melanin in a relatively short time, and make the skin more shiny, bright, smooth and delicate. DETAILED DESCRIPTION

[0019] The invention provides a skin whitening, rejuvenating and brightening composition, comprising the following components in parts by weight: 10-20 parts of niacinamide, 10-20 parts of phenethylresorcinol nanoparticles, 8-12 parts of arbutin, 5-10 parts of lilac flower oil, 5-10 parts of pearl extract, 10-20 parts of purslane extract, 3-8 parts of allantoin and 3-8 parts of trehalose.

[0020] The niacinamide of the present invention can inhibit the transfer of melanin to epidermal cells, reduce pigmentation, brighten skin tone, and improve dullness; increase the production of collagen in the skin, improve the elasticity and firmness of the skin, reduce the appearance of wrinkles, prevent skin moisture loss, and keep the skin moisturized.

[0021] The preparation method of the phenethylresorcinol nanoparticles of the present invention preferably comprises: dissolving phospholipids in methanol, adding a cyclodextrin aqueous solution and stirring, removing methanol to obtain an aqueous phase; dissolving phenethylresorcinol in anhydrous ethanol, adding the aqueous phase, removing ethanol, and freeze-drying to obtain the phenethylresorcinol nanoparticles.

[0022] The phospholipids of the present invention preferably include one or two of soybean lecithin, hydrogenated soybean lecithin or egg yolk lecithin, more preferably hydrogenated soybean lecithin; the cyclodextrin is preferably water-soluble cyclodextrin, more preferably hydroxypropyl-β-cyclodextrin.

[0023] The mass volume ratio of the phospholipid to methanol of the present invention is 8-12:1 mg / mL, more preferably 10:1 mg / mL; the mass volume ratio of cyclodextrin to water in the cyclodextrin aqueous solution is preferably 1-3:1-3 mg / mL, more preferably 2:2 mg / mL; the stirring speed is preferably 400-600 rpm, more preferably 500 rpm; the mass volume ratio of phenethyl resorcinol to methanol is preferably 15-25 mg / mL, more preferably 20 mg / mL; the mass ratio of the phospholipid, cyclodextrin and phenethyl resorcinol is preferably 1:2-4:1-3, more preferably 1:3:2.

[0024] Phenylethyl resorcinol can reduce the production of melanin and reduce skin coloration caused by ultraviolet radiation. However, there are the following problems in its application: the solubility of phenethyl resorcinol in water is low, and its content is too low when used in cosmetic formulas, making it difficult to achieve a whitening effect. In addition, phenethyl resorcinol is photo-instable and will rapidly change from white to beige and finally to pink under light conditions, causing the phenethyl resorcinol content to drop rapidly, affecting the appearance of the cosmetics and the bioavailability of phenethyl resorcinol.

[0025] The phospholipids of the present invention enable it to self-assemble in the solution to form a lipid bilayer structure, providing a stable structure for the nanoparticles; have good biocompatibility, reduce the toxicity and immune response of the nanoparticles to the organism; improve the drug loading efficiency of the nanoparticles; can delay the release of phenylethyl resorcinol, can inhibit the skin coloring caused by ultraviolet irradiation of the skin for a long time, and achieve a long-lasting effect; can help the nanoparticles specifically target phenylethyl resorcinol to melanocytes, inhibit melanocytes from producing melanin, and then achieve a whitening effect. In addition, phospholipids can form a protective film on the surface of the skin to prevent water loss, enhance the skin's moisturizing ability, and keep the skin moist and soft; help repair damaged skin barriers, enhance the skin's resistance, and reduce the damage to the skin caused by external stimuli. The cyclodextrins of the present invention can improve the solubility of phenylethyl resorcinol in water; enhance the physical stability of nanoparticles, prevent nanoparticles from aggregating and precipitating; improve the solubility and permeability of the drug, make it easier to be absorbed and utilized by the body, and improve the bioavailability of the drug. In addition, the cyclodextrin of the present invention can protect phenethyl resorcinol from being oxidized, thereby improving the whitening effect of phenethyl resorcinol. The preparation method of phenethyl resorcinol nanoparticles of the present invention can encapsulate phenethyl resorcinol, thereby improving the solubility and stability, thereby improving the whitening effect of phenethyl resorcinol.

[0026] The arbutin described in the present invention can effectively inhibit the activity of tyrosinase, reduce the production of melanin, thereby reducing skin pigmentation, making the skin color even and fair, and fading spots such as freckles and chloasma; it can resist the damage of free radicals to the skin, slow down the speed of skin aging, and keep the skin healthy; it helps to reduce the inflammatory response of the skin, and has a certain soothing effect on acne skin or skin sensitivity and inflammation; it can reduce the damage of ultraviolet rays to the skin to a certain extent and prevent excessive production of melanin.

[0027] The lilac flower oil of the present invention can promote blood circulation of the skin, enhance the penetration of whitening ingredients into the deep layers of the skin, and thus improve the whitening effect.

[0028] The preparation method of the pearl extract of the present invention preferably comprises: crushing the pearls and mixing them with glucose, inoculating composite microorganisms and fermenting for 24-36 hours, filtering to obtain the fermentation liquid, adding neutral protease for enzymatic hydrolysis, filtering under ultra-high pressure, and freeze-drying to obtain the pearl extract.

[0029] The particle size of the pearl powder crushed in the present invention is preferably 0.3-1 μm, and the obtained pearl powder has a higher specific surface area and better solubility, which is more conducive to the subsequent fermentation and enzymatic hydrolysis steps.

[0030] When the pearl powder of the present invention is mixed with glucose, water is also added; the mass ratio of the pearl powder to glucose is preferably 1:1-3:8-12, more preferably 1:2:10.

[0031] The composite microorganism of the present invention is preferably Bacillus tequila and Fusarium venezia, more preferably, the Bacillus tequila is Bacillus tequila S-37, with a deposit number of CCTCC NO: M2021205, which has been disclosed in the patent with application number 202110359359.8; the Fusarium venezia is Fusarium venezia TB02, with a deposit number of CGMCCNO.23057, which has been disclosed in the patent with application number 202310017398.9. The Bacillus tequila and Fusarium venezia of the present invention can decompose the macromolecular protein in the pearl powder into small molecular proteins, which can not only improve the antioxidant activity of the fermentation broth, but also greatly increase the amino acid content in the fermentation broth.

[0032] The composite microorganism of the present invention is inoculated into pearl powder and glucose solution in the form of seed liquid; the volume ratio of the tequila bacillus seed liquid to the venezuelae Fusarium seed liquid is 1-3:1-3; the inoculation amount is preferably 2%-8%, more preferably 5%; the fermentation temperature is preferably 28-35°C, more preferably 30°C.

[0033] The mass ratio of the neutral protease to the pearl powder of the present invention is preferably 1:100-200, more preferably 1:150; the pH value of the enzymatic hydrolysis is preferably 6.0-7.5, more preferably 7.0, the temperature is preferably 45-55°C, more preferably 50°C, the time is preferably 2-5h, more preferably 3h; the filter membrane of the ultra-pressure filtration is preferably 0.45μm.

[0034] The preparation method of the pearl extract of the present invention combines fermentation and enzymatic hydrolysis steps, which can fully decompose the pearl protein into small molecule proteins or peptides, making it easier for the skin to absorb, thereby improving and reducing the generation and deposition of melanin in a shorter period of time.

[0035] The preparation method of the purslane extract of the present invention preferably comprises: drying and crushing the purslane, performing ethanol microwave extraction, filtering and freeze-drying to obtain the purslane extract. The volume percentage of the ethanol is preferably 60%-70%, more preferably 65%, the solid-liquid ratio is 1:8-12g / mL, more preferably 1:10, the frequency is preferably 15-30kHz, more preferably 20kHz, the temperature is preferably 45-55°C, more preferably 50°C, and the time is preferably 60-90min, more preferably 75min.

[0036] The purslane extract prepared by the present invention has anti-inflammatory, antioxidant and anti-irritation properties, can reduce the inflammatory response of the skin, and relieve the discomfort symptoms of the skin such as redness, swelling, itching, burning, etc., thereby achieving the effect of soothing the skin.

[0037] The allantoin of the present invention can increase the moisture content of the skin, keep the skin moisturized, improve dry and rough skin texture; accelerate cell proliferation and the generation of new tissues; reduce the inflammatory response of the skin, play a soothing and calming role, and relieve the redness, swelling, itching and pain of the skin; make the skin stratum corneum soft, promote the shedding of aged keratin, and make the skin smoother.

[0038] The trehalose of the present invention can form a protective film on the cell surface to prevent water loss and keep the skin moisturized; protect the structure and function of biological macromolecules such as proteins, lipids and nucleic acids, and enhance the stress resistance of cells; reduce the damage of free radicals to the skin and delay skin aging; protect skin cells from damage; help repair and strengthen the skin barrier and improve the skin's resistance.

[0039] The present invention also provides a method for preparing the above-mentioned skin whitening, rejuvenating and brightening composition, comprising: mixing and stirring the auxiliary materials with the niacinamide, phenylethylresorcinol nanoparticles, arbutin, lilac flower oil, pearl extract, purslane extract, allantoin and trehalose to obtain the composition.

[0040] The auxiliary materials of the present invention include emulsifier, emollient, moisturizer, thickener, preservative, essence and deionized water.

[0041] The composition of the present invention is preferably a lotion, emulsion or cream.

[0042] The present invention also provides application of the skin whitening, rejuvenation and brightening composition in the preparation of cosmetics.

[0043] In the present invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0044] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0045] The preparation method of the composite microbial seed solution in the following embodiment is as follows:

[0046] Preparation of seed solution of Bacillus tequila S-37: inoculate Bacillus tequila S-37 onto a slant culture medium and culture at 30-35°C for 24 hours to obtain activated Bacillus tequila S-37; inoculate the activated Bacillus tequila S-37 into a seed solution culture medium and culture until the bacterial solution density reaches 2.5×10 8 CFU / mL, and the seed liquid of Bacillus tequila S-37 was obtained.

[0047] Preparation of Venezia Fusarium TB02 seed solution: inoculate Venezia Fusarium TB02 onto a slant culture medium and culture at 28°C-32°C for 36 h to obtain activated Venezia Fusarium TB02; inoculate the activated Venezia Fusarium TB02 into a seed culture medium and culture until the bacterial solution density reaches 2.5×10 8 CFU / mL, and the seed liquid of Fusarium venezuelae TB02 was obtained.

[0048] The above-mentioned Bacillus tergillosis S-37 seed solution and the Fusarium venezuelae TB02 seed solution were mixed in equal volumes to obtain a composite microbial seed solution.

[0049] Example 1

[0050] Preparation of skin whitening, rejuvenation and brightening composition (toner)

[0051] (1) Preparation of phenylethylresorcinol nanoparticles

[0052] 10 mg of hydroxypropyl-β-cyclodextrin was mixed with 1 mL of methanol, and then added to a hydrogenated soybean lecithin aqueous solution (mixed with 30 mg of hydrogenated soybean lecithin and 30 mL of water), stirred and mixed evenly, and methanol was removed by rotary evaporation to obtain an aqueous phase; a phenethylresorcinol ethanol solution (20 g of phenethylresorcinol was dissolved in 100 mL of anhydrous ethanol) was injected into the aqueous phase under a stirring condition of 500 rpm, and after rotary evaporation to remove ethanol, the solution was mixed with 10 mg of trehalose, and freeze-dried to obtain the phenethylresorcinol nanoparticles;

[0053] (2) Preparation of pearl extract

[0054] The pearls are crushed to 0.5 μm, 10 g of pearl powder, 20 g of glucose and 100 g of water are mixed, and then 5% by volume of composite microbial seed solution is inoculated, and fermented at 30° C. for 30 hours, filtered to obtain a fermentation liquid, 65 mg of neutral protease is added, and enzymolysis is carried out at pH 7.0 and 50° C. for 3 hours, filtered through a 0.45 μm membrane under ultra-pressure, 30 mg of trehalose is added, and freeze-dried to obtain the pearl extract;

[0055] (3) Preparation of Portulaca Oleracea Extract

[0056] Purslane is dried and crushed, passed through a 200-mesh sieve, 10 g of purslane powder is mixed with 100 mL of 65% by volume ethanol, microwave-extracted for 75 min at 20 kHz and 50° C., filtered, 30 mg of trehalose is added, and freeze-dried to obtain the purslane extract;

[0057] (4) Preparation of skin whitening and brightening composition

[0058] Accurately weigh 15 g of niacinamide, 15 g of phenethylresorcinol nanoparticles, 10 g of arbutin, 7 g of clove oil, 8 g of pearl extract, 15 g of purslane extract, 5 g of allantoin and 5 g of trehalose, mix them evenly to obtain effective ingredients, and mix them with 300 g of auxiliary materials to obtain the skin whitening, rejuvenation and brightening composition;

[0059] The auxiliary materials are: 18 g of sorbitol, 4 g of carbomer-934 and 278 g of deionized water.

[0060] Example 2

[0061] Preparation of skin whitening and brightening composition (emulsion)

[0062] The specific implementation is the same as that of Example 1, except that in the preparation of the skin whitening and brightening composition in step (4), the auxiliary materials are: 18 g glycerin, 1.7 g disodium EDTA, 0.3 g potassium sorbate, 1 g vitamin E, 6 g polyvinyl alcohol-2488 and 273 g deionized water.

[0063] Example 3

[0064] Preparation of skin whitening, rejuvenation and brightening composition (face cream)

[0065] The specific implementation method is the same as that of Example 1, except that in step (4), the skin whitening and brightening composition is prepared, the auxiliary materials are 50g of dimethyl silicone oil, 35g of glycerin, 2g of vitamin C, 3g of phenoxyethanol, 35g of soybean lecithin, 25g of gelatin and 150g of deionized water.

[0066] Comparative Example 1

[0067] The specific implementation is the same as that of Example 1, except that the phenethylresorcinol nanoparticles are replaced with phenethylresorcinol.

[0068] Comparative Example 2

[0069] The specific implementation method is the same as that of Example 1, except that (2) the preparation of pearl extract: the pearls are crushed to 0.5 μm, 10 g of the pearls are mixed with 100 g of water, 65 mg of neutral protease is added, and enzymatic hydrolysis is carried out at pH 7.0 and 50° C. for 3 h, and the pearl extract is filtered through a 0.45 μm membrane under ultra-high pressure, 30 mg of trehalose is added, and the pearl extract is freeze-dried to obtain the pearl extract.

[0070] Comparative Example 3

[0071] The specific implementation method is the same as that of Example 1, except that (2) the preparation of pearl extract: the pearls are crushed to 0.5 μm, 10 g of pearl powder, 20 g of glucose and 100 g of water are mixed, inoculated with 5% composite microbial seed liquid, fermented at 30° C. for 30 h, filtered through a 0.45 μm membrane under ultra-pressure, 30 mg of trehalose is added, and freeze-dried to obtain the pearl extract.

[0072] Comparative Example 4

[0073] The specific implementation method is the same as that of Example 1, except that (3) Preparation of purslane extract: Purslane is dried and crushed, passed through a 200-mesh sieve, 10 g of purslane powder is mixed with 100 mL of water, and microwave extracted at 20 kHz and 50° C. for 75 min. After filtering, 30 mg of trehalose is added, and freeze-dried to obtain the purslane extract.

[0074] Comparative Example 5

[0075] The specific implementation method is the same as that of Example 1, except that (4) no phenethyl resorcinol nanoparticles are added in the preparation of the skin whitening, rejuvenation and brightening composition.

[0076] Comparative Example 6

[0077] The specific implementation is the same as that of Example 1, except that (4) pearl extract is not added in the preparation of the skin whitening, rejuvenation and brightening composition.

[0078] Comparative Example 7

[0079] The specific implementation method is the same as that of Example 1, except that (4) niacinamide, phenethylresorcinol nanoparticles, arbutin and pearl extract are not added in the preparation of the skin whitening, rejuvenation and brightening composition.

[0080] Comparative Example 8

[0081] The specific implementation method is the same as that of Example 1, except that (4) no clove flower oil, purslane extract, allantoin and trehalose are added in the preparation of the skin whitening, rejuvenation and brightening composition.

[0082] Comparative Examples 5-8 were carried out with the ratio of active ingredient to auxiliary material unchanged.

[0083] Experimental Example 1

[0084] Inhibitory effect of skin whitening, rejuvenation and brightening composition on tyrosinase in vitro

[0085] Tyrosinase is a key enzyme in the synthesis of melanin. By testing the ability of the composition to inhibit tyrosinase activity, its potential whitening effect can be preliminarily evaluated.

[0086] Tyrosinase activity detection experiment refers to Wei Chuyuan's "4-butylresorcinol microemulsion gel preparation and whitening antioxidant research" tyrosinase inhibition test: prepare 100U / mL tyrosinase solution and 1mg / mL L-tyrosine solution respectively. The compositions in Example 1 and Comparative Examples 1-8 (containing only active ingredients, excluding excipients) were prepared into 0.05, 0.1, 0.5, 1.0, 1.5, 3.0, 5.0, 10.0, 50.0, 300.0μg / mL solutions with PBS buffer respectively.

[0087] Take a group of EP tubes (A3), add 0.1mL of the test solution of different mass concentrations, 0.2mL of L-tyrosine solution with a mass fraction of 0.1%, add PBS buffer to 0.6mL, and mix; take another group of EP tubes (A4), add 0.1mL of the test solution of different mass concentrations, add PBS buffer to 0.6mL, and mix. Take another EP tube (A1), add 0.2mL of L-tyrosine solution with a mass fraction of 0.1%, add PBS buffer to 0.6mL; take an EP tube (A2), add PBS buffer to 0.6mL. Immediately add 0.1mL of tyrosinase solution, place in a 37℃ oven for reaction for 40min, and then quickly transfer to a 96-well plate, 200μL per well, use an enzyme reader to measure the absorbance at 475nm, and calculate the tyrosinase inhibition rate according to formula (1). The specific results are shown in Table 1.

[0088] Tyrosinase inhibition rate (%) = [(A1-A2)-(A3-A4)] / (A1-A2)×100% (1);

[0089] Wherein, A1 is the absorbance of the blank control group containing tyrosine, A2 is the absorbance of the blank control substance not containing tyrosine, A3 is the absorbance of the test substance group containing tyrosine, and A4 is the absorbance of the test substance group not containing tyrosine.

[0090] Table 1 Tyrosinase inhibition rate of different concentrations of the composition (%)

[0091]

[0092]

[0093] The results of Table 1 show that as the concentration of the active ingredient in each group increases, the tyrosinase inhibition rate increases significantly. At low concentrations, the tyrosinase inhibition rate of Comparative Example 1 is higher than that of Example 1 because the reaction time between the active ingredient and tyrosinase is only 40 minutes, while the carrier in Example 1 can continuously and continuously release phenylethyl resorcinol, thereby prolonging the action time of phenylethyl resorcinol. According to the data of Example 1 and Comparative Examples 2-3, when the pearl extract prepared by the fermentation + enzymolysis method is used in combination with other ingredients, the tyrosinase inhibition rate can be significantly increased, thereby improving the whitening effect. According to the data of Example 1 and Comparative Examples 5-7, the whitening active ingredient added by the present invention can significantly inhibit the tyrosinase and improve the whitening effect. According to the data of Example 1 and Comparative Example 8, the lilac oil, purslane extract, allantoin and trehalose added by the present invention have the effect of promoting the whitening ingredient to inhibit tyrosinase. From the above results, it can be seen that there is a synergistic effect between the components of the present invention, achieving a significant tyrosinase inhibition rate, thereby improving the whitening effect.

[0094] Experimental Example 2

[0095] Effects of skin whitening, rejuvenation and brightening composition on melanin in melanocytes

[0096] 1. Cell culture

[0097] Mouse melanoma B16F10 cells were grown in RPMI-1640 complete medium (containing 10% fetal bovine serum and 1% double antibody) at 37°C and 5% CO. 2 The culture was carried out in a saturated humidity environment.

[0098] When the cells grow to the logarithmic phase, wash the cells with PBS, add 1.5mL 0.25% trypsin and place in the incubator for digestion for 1min, aspirate the trypsin and add 3mL culture medium and gently blow to prepare a cell suspension. Take an appropriate amount of cells and inoculate them in a cell plate and place in the incubator for culture.

[0099] 2. Determination of cell proliferation inhibition rate

[0100] The density of mouse melanoma B16F10 cells was adjusted to 2.0 × 10 per ml. 5, inoculated in a 96-well plate, and cultured for 12 hours. The compositions in Example 1 and Comparative Examples 1-8 (containing only active ingredients, excluding excipients) were added to the complete medium at final concentrations of (0, 3.0, 5.0, 10.0, 50.0, 300.0 μg / mL), respectively, and added to the attached cells, 100 μL per well, and an equal volume of culture medium was set as a control group, with 6 replicates per group. After incubation for 48 hours, 10 μL of CCK8 solution was added to each well, placed in an incubator and incubated for 30 minutes, and the absorbance (A) value of each well was detected at 450 nm using an ELISA reader, and the cell proliferation inhibition rate was calculated according to formula (2).

[0101] Cell proliferation inhibition rate (%) = [1-(A treatment group-A blank well) / (A control group-A blank well)] × 100% (2).

[0102] Table 2 Inhibition rate of proliferation of B16F10 cells by different concentrations of the composition (%)

[0103]

[0104]

[0105] The results of Table 2 show that with the increase of the concentration of the effective ingredients in each group, the proliferation inhibition rate of B16F10 increases significantly. Compared with Example 1, Comparative Example 1 has a high proliferation inhibition rate of B16F10 cells because phenethyl resorcinol is more irritating to B16F10 cells than the phenethyl resorcinol nanoparticles of the present invention. According to the data of Example 1 and Comparative Examples 2-3, the pearl extract prepared by the fermentation + enzymolysis method can inhibit the proliferation of B16F10 cells when used in combination with other ingredients. According to the data of Example 1 and Comparative Examples 5-7, the whitening effective ingredients added by the present invention can inhibit the proliferation of B16F10 cells. According to the data of Example 1 and Comparative Example 8, the lilac oil, purslane extract, allantoin and trehalose added by the present invention have the effect of promoting the whitening ingredients to inhibit the proliferation of B16F10 cells. From the above results, it can be seen that the components of the present invention have a synergistic effect and can inhibit the proliferation of B16F10 cells.

[0106] 3. Effect of the whitening, rejuvenation and brightening composition on melanin synthesis in B16F10 cells

[0107] Select B16F10 cells in the logarithmic growth phase and add 2.0×10 5The cells were inoculated in a 6-well culture plate (2 mL per well), and 100 μM α-MSH and working solutions of the compositions in Example 1 and Comparative Examples 1-8 (containing only active ingredients, excluding excipients) with final concentrations of (0, 3.0, 5.0, 10.0, 50.0, 300.0 μg / mL) were added and cultured in an incubator for 72 h. After incubation, the cells were washed twice with ice PBS, and the cells were collected by trypsin digestion. 100 μL of 0.1 mol / L potassium phosphate buffer (pH=6.8) containing 1% Triton X-100 was added to each tube and the cells were lysed in an ice bath for 1 hour. The supernatant was then collected in a new tube for BCA protein and tyrosinase activity measurement. 500 μL of 1 mol / L NaOH solution containing 10% DMSO was added to the remaining black cell precipitate in each tube and incubated in an 80°C water bath for 1 hour. After incubation, the cells were centrifuged at 3000 r / min for 5 minutes (radius of 5 cm), and 200 μL of the supernatant was transferred to a 96-well plate to determine the melanin content at 475 nm. The relative content of melanin was determined according to formula (3). The specific results are shown in Table 3.

[0108] Melanin relative content (%) = 1 - (A sample / A blank) × 100% (3);

[0109] Wherein, blank refers to the addition of only B16F10 cells in the logarithmic growth phase.

[0110] After testing, the relative content of melanin was 170% when only 100 μM α-MSH was added without adding the composition, and the intracellular melanin content increased significantly, indicating that the melanin high-expressing cell model was successfully established.

[0111] Table 3 Effect of different concentrations of the composition on the relative content of melanin in B16F10 cells (%)

[0112]

[0113] The results of Table 3 show that as the concentration of each group of active ingredients increases, the relative content of melanin in B16F10 cells shows a downward trend. Compared with Comparative Example 1, the relative content of melanin in Example 1 is lower because the phenethyl resorcinol nanoparticles of the present invention can continuously release phenethyl resorcinol due to the encapsulation of the carrier, and can act on B16F10 cells for a long time, reducing and inhibiting the production of melanin. According to the data of Example 1 and Comparative Examples 2-3, it can be seen that the pearl extract prepared by the fermentation + enzymolysis method can further reduce the content of melanin in B16F10 cells when used in combination with other ingredients. According to the data of Example 1 and Comparative Examples 5-7, it can be seen that the whitening active ingredients added by the present invention can further reduce the content of melanin in B16F10 cells. According to the data of Example 1 and Comparative Example 8, the lilac oil, purslane extract, allantoin and trehalose added by the present invention have the effect of promoting whitening ingredients to inhibit the production of melanin in B16F10 cells. From the above results, it can be seen that the components of the present invention have a synergistic effect and can reduce the content of melanin in B16F10 cells.

[0114] Experimental Example 3

[0115] Evaluation of the efficacy of the composition on whitening, rejuvenation and brightening of human skin

[0116] 1. Human skin patch test

[0117] According to the requirements of the closed patch test in the "Technical Specification for Safety of Cosmetics", human skin patch tests were performed on the compositions of Examples 1-3 and Comparative Examples 1-8.

[0118] A total of 360 subjects aged 35-50 years (without chronic diseases, major diseases or allergies, and not taking any medications, with a male to female ratio of 1:1) were selected and divided into 12 groups on average. There were no significant differences among the groups.

[0119] The human skin patch test was performed on the above subjects: gauze soaked with lotions of Examples 1-3 and Comparative Examples 1-8 was applied to the normal skin on the inner side of the forearm of the subject, then covered with cellophane and fixed with gauze, and the results of the inner forearm skin were observed after 24-72 hours. A blank control (without any treatment) was used as a negative control.

[0120] The experimental results show that during the 0-72h patch test, except for Comparative Example 1 (the score levels at 24h and 72h were both 2 weak positive reactions (erythema reaction); erythema, infiltration, edema, and papules may be present), no positive reactions occurred in any of the groups (the reaction results were all negative reactions 0), that is, the lotions of Examples 1-3 and Comparative Examples 2-8 did not cause adverse reactions to the skin.

[0121] Compared with Comparative Example 1, Example 1 of the present invention uses phenethyl resorcinol nanoparticles, and uses a suitable carrier to encapsulate phenethyl resorcinol, thereby reducing the irritation of phenethyl resorcinol to the skin. It can be seen that the composition of the present invention is mild and non-irritating, and will not cause adverse reactions to the skin.

[0122] 2. Evaluation of human skin whitening, rejuvenation and brightening effects

[0123] The compositions of Examples 1-3 and Comparative Examples 2-8 were used as test samples, and the excipients in each group were used as placebos, and a double-blind, open, randomized, self-controlled before-after observation study was conducted.

[0124] 300 subjects were selected from 1 and randomly divided into 10 groups (no significant difference between the groups). Each group of subjects applied the test sample on one side of the face and the placebo on the other side for 28 days, twice a day, once in the morning and once in the evening. Facial tests were performed on the 0th, 14th and 28th days to evaluate the whitening, tenderizing and brightening effects of the product.

[0125] The skin color probe of the skin analyzer was used to detect and record the skin brightness ITA° values ​​of the specified parts of the subject's face, and the improvement rate of ITA° was calculated according to formula (4). The specific results are shown in Table 4.

[0126] ITA° improvement rate (%) = (ITA° value on day N - ITA° value on day 0) / ITA° value on day 0 × 100% (4); wherein N is 14 or 28.

[0127] ITA° refers to the individual type angle, which is a parameter that characterizes human skin color by measuring the skin Lab color space data through a skin colorimeter or a reflectance spectrophotometer. The larger the ITA° value, the lighter the skin color, and vice versa, the darker the skin color.

[0128] Table 4 Improvement rate of skin brightness ITA° in each group (%)

[0129] Group Day 14 Day 28 Example 1 8.6 38.3 Example 2 9.5 36.5 Example 3 8.9 39.1 Comparative Example 2 4.3 20.2 Comparative Example 3 4.0 19.3 Comparative Example 4 3.9 21.7 Comparative Example 5 3.0 14.3 Comparative Example 6 2.8 13.7 Comparative Example 7 0.9 3.5 Comparative Example 8 1.6 10.1

[0130] As can be seen from the results in Table 4, each group has a positive effect on improving the brightness of the skin. Examples 1-3 of the present invention have been significantly improved after 14 days of use, and the effect is better than that of comparative examples 2-8. Therefore, the composition of the present invention is highly effective and mild, and can achieve the effect of whitening, rejuvenating and brightening the skin in a short time.

[0131] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A skin whitening and brightening composition, characterized in that: The invention comprises the following components in parts by weight: 10-20 parts of niacinamide, 10-20 parts of phenethylresorcinol nanoparticles, 8-12 parts of arbutin, 5-10 parts of lilac flower oil, 5-10 parts of pearl extract, 10-20 parts of purslane extract, 3-8 parts of allantoin and 3-8 parts of trehalose; The preparation method of the phenethyl resorcinol nanoparticles comprises: dissolving phospholipids in methanol, adding a cyclodextrin aqueous solution and stirring, removing methanol to obtain a water phase; dissolving phenethyl resorcinol in anhydrous ethanol, adding the anhydrous ethanol to the water phase, removing ethanol, and freeze-drying to obtain the phenethyl resorcinol nanoparticles; The preparation method of the pearl extract comprises: crushing the pearls and mixing them with glucose, inoculating composite microorganisms for fermentation for 24-36 hours, filtering to obtain a fermentation liquid, adding neutral protease for enzymolysis, filtering under ultra-high pressure, and freeze-drying to obtain the pearl extract; the composite microorganisms are Bacillus tequila and Fusarium venezuelae; The preparation method of the purslane extract comprises: drying and crushing the purslane, performing ethanol microwave extraction, filtering and freeze-drying to obtain the purslane extract; the volume percentage of the ethanol is preferably 60%-70%.

2. The skin whitening and brightening composition according to claim 1, characterized in that: The phospholipids include one or two of soybean lecithin, hydrogenated soybean lecithin or egg yolk lecithin; and the cyclodextrin is water-soluble cyclodextrin.

3. The skin whitening and brightening composition according to claim 2, characterized in that: The mass ratio of the phospholipid, cyclodextrin and phenethylresorcinol is 1:2-4:1-3.

4. A method for preparing the skin whitening, rejuvenation and brightening composition according to any one of claims 1 to 3, characterized in that: include: The auxiliary materials are mixed and stirred with the niacinamide, phenylethyl resorcinol nanoparticles, arbutin, lilac flower oil, pearl extract, purslane extract, allantoin and trehalose to obtain the composition.

5. The preparation method according to claim 4, characterized in that: The auxiliary materials include emulsifier, emollient, moisturizer, thickener, preservative, essence and deionized water.

6. Use of the skin whitening, rejuvenation and brightening composition according to claims 1 to 3 or the skin whitening, rejuvenation and brightening composition obtained by the preparation method according to any one of claims 4 to 5 in the preparation of cosmetics.

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