A whitening composition, preparation method and use

Abstract

The invention belongs to the technical field of skin care, and discloses a whitening composition, a preparation method and a use. The whitening composition comprises, by weight percentage, 8-25% of a white mulberry extract, 0.1-10% of a large-flowered cactus flower extract, 0.1-10% of a eucystis extract, 0.1-5% of decarboxylated carnosine hydrochloride, and 0.1-5% of cyclodextrin, and the balance is water. The invention discloses a whitening composition, which takes white mulberry as a main whitening component, is supplemented with eucystis, decarboxylated carnosine hydrochloride, and a large-flowered cactus flower extract for moisturizing, anti-inflammatory, anti-aging, and skin tightening effects, and comprehensively regulates the skin state while synergistically improving the whitening effect of the skin.

Description

Technical Field

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

[0002] As people pay more and more attention to the management of their appearance, whitening and translucent skin has also become a point that people pay more attention to. Skin whitening care is a research hotspot in the field of skin care.

[0003] Current skin care products mainly achieve the purpose of whitening from different angles by adding different functional ingredients.

[0004] For example, prior art 1: CN109893464A discloses a whitening and freckle-removing skin care product that blocks, inhibits and reduces melanin and a preparation method thereof, and in particular relates to a whitening and freckle-removing formula that blocks, inhibits and reduces melanin. The formula consists of niacinamide, betaine, β-glucan, iris extract, mulberry bark extract, filtrate of fermentation product of coated yeast, spirulina extract, chlorella extract, rice germ extract, barley malt extract, wheat malt extract, etc., and can block, inhibit and reduce melanin, accelerate cell metabolism, and speed up the shedding of melanin keratinocytes; eliminate skin aging, inhibit, delay and reduce the formation of wrinkles, and promote skin regeneration and skin conditioning effects.

[0005] Prior art 1 uses various components to whiten the skin from the perspective of inhibiting melanin formation and accelerating the shedding of melanin keratinocytes, while supplemented with anti-aging components to regulate the skin condition to achieve a comprehensive skin care effect.

[0006] Similarly, prior art 2 also uses the same skin care mechanism as prior art 1 to improve the whitening effect of the skin. Prior art 2: CN103919714B discloses a whitening composition, which is composed of the following components in mass percentage: niacinamide: 1-20%, arbutin: 1-5%, carnitine: 1-10%, angelica root extract: 0.1-1%, mulberry bark extract: 0.1-1%, peony root extract: 0.1-1%, Bletilla striata root extract: 0.1-1%, Poria root extract: 0.01-0.5%, Atractylodes macrocephala root extract: 0.01-0.5%, pearl extract: 0.01-0.5%, glycerin: 10-50%, and the balance is water.

[0007] Existing technology 2 uses niacinamide, carnitine, mulberry bark extract, pearl extract and other components to achieve whitening effects through mechanisms such as inhibiting melanin formation, inhibiting the transfer of melanin particles to keratinocytes, and accelerating the shedding of the stratum corneum. At the same time, it is supplemented with anti-aging components such as angelica root extract, peony root extract, and atractylodes root extract, and dark spot removal components such as bletilla root extract and Poria root extract to effectively synergize whitening.

[0008] It can be seen from the prior art 1 and the prior art 2 that the technical routes of the prior art are to compound components of whitening mechanisms such as inhibiting melanin formation, and then supplemented with anti-aging components to regulate the skin condition to further promote the whitening effect of skin care products, and the whitening effects they show are good; however, the regulation of skin condition by the prior art is not comprehensive enough, resulting in the synergistic improvement of skin whitening by skin care products is not high.

[0009] Therefore, the technical problem to be solved by the present invention is: how to compound a whitening composition that can effectively whiten the skin while comprehensively regulating the skin condition, and effectively and synergistically enhance the whitening effect of the composition. Summary of the invention

[0010] One of the purposes of the present invention is to provide a whitening composition that can comprehensively regulate the state of the skin, effectively improve the overall smoothness of the skin, and synergistically block, inhibit, and reduce melanin with whitening components, fully inhibiting tyrosinase activity while being able to inhibit B16 cell melanin.

[0011] Another object of the present invention is to provide a method for preparing a whitening composition, wherein the whitening composition prepared by the preparation method can maximize the whitening effect of the composition.

[0012] At the same time, the present invention also provides a use of a whitening composition, which can be used to prepare cosmetics or skin care products with whitening effects.

[0013] To achieve the above object, the present invention provides a whitening composition, which comprises, by weight percentage, 8-25% of a Bletilla striata extract, 0.1-10% of a large-flowered cactus flower extract, 0.1-10% of a Eucystis salsa extract, 0.1-5% of decarboxylated carnosine hydrochloride, 0.1-5% of cyclodextrin, and the balance is water.

[0014] As a traditional Chinese medicinal material, Baiwei has been recorded in herbal works of all dynasties, such as "Compendium of Materia Medica" and "Ming Yi Bie Lu", and has a long history of use. Experimental studies have shown that the polyphenols rich in Baiwei can delay the skin aging process and improve the damage caused by skin exposure to ultraviolet rays. At the same time, it can inhibit the production of melanin and has the effect of whitening and removing spots.

[0015] Streptospira oleifera is a brown algae that grows in freshwater and saltwater environments. It has high nutritional value and multiple skin care benefits such as moisturizing, anti-oxidation, anti-inflammatory, and promoting skin cell regeneration.

[0016] Decarboxylated carnosine hydrochloride is a decarboxylated peptide in the form of hydrochloride salt, which has specific biological activity and mechanism of action. In the field of skin care, decarboxylated carnosine hydrochloride is usually used as an ingredient for anti-aging and skin repair. It is a peptide substance that can promote collagen synthesis and enhance skin elasticity. Decarboxylated peptide in the form of hydrochloride salt is more easily absorbed and utilized by the skin; therefore, decarboxylated carnosine hydrochloride can help reduce wrinkles and increase skin firmness, thereby improving skin texture and appearance, and playing an auxiliary whitening role.

[0017] The large-flowered cactus flower extract contains a variety of antioxidants that can help reduce the damage of free radicals to the skin. Combined with the inhibitory effect of cactus extract on the activity of androgen receptors, it can be used in acne cosmetics and has a preventive and therapeutic effect on acne caused by high androgen levels. At the same time, cactus extract can promote the production of skin growth factor and has a skin rejuvenating effect.

[0018] The present invention also discloses a method for preparing the whitening composition, comprising the following steps:

[0019] Step 1: extracting the white osmanthus, the flower of Opuntia grandiflora, and the Streptospira oleracea to obtain the white osmanthus extract, the flower of Opuntia grandiflora, and the Streptospira oleracea extract;

[0020] Step 2: stirring and dissolving the white mulberry extract, the large-flowered cactus flower extract, the Eucystis extract, decarboxylated carnosine hydrochloride, cyclodextrin and water at a temperature of 40-50° C. for 20-30 minutes to obtain a mixed solution;

[0021] Step 3: The mixed solution is subjected to ultrafiltration and sterilization treatment in sequence to obtain the whitening composition.

[0022] Furthermore, the Bletilla striata extract in step 1 is obtained by extracting through the following steps:

[0023] Step a: crush the white mulberry into powder;

[0024] Step b: adding 60-80% ethanol solution with a mass concentration of 10-20 times the weight of the powder, stirring and extracting at a temperature of 48-52° C. for 2-4 hours, and then centrifuging and filtering to obtain a filtrate;

[0025] Step c: Concentrate the filtrate to a weight of 2-4 times the weight of the concentrated juice powder to obtain a iris extract.

[0026] Furthermore, the large-flowered cactus flower extract in step 1 is obtained by extracting through the following steps:

[0027] Step d: putting the large-flowered cactus flowers into a homogenizer, adding water twice the weight of the large-flowered cactus flowers for homogenization, and filtering to obtain a filtrate;

[0028] Step e: adding water 10-14 times the weight of the large-flowered cactus flower to the filtrate, stirring and extracting at a temperature of 65-75° C. for 2-4 hours, and then centrifuging to obtain a supernatant;

[0029] Step f: Concentrating the supernatant to 2-4 times the weight of the large-flowered cactus flower to obtain a large-flowered cactus flower extract.

[0030] Furthermore, the Eucystis extract in step 1 is obtained by extracting through the following steps:

[0031] Step g: breaking the cell wall of Eucystis spp. to obtain a broken cell wall product;

[0032] Step h: adding water 10-14 times the weight of Eucystis to the broken cell wall material, inactivating the enzyme at 65-75°C for 30-60 minutes, cooling to 45-55°C, stirring and extracting for 2-4 hours, and then centrifuging to obtain a supernatant;

[0033] Step i: Concentrate the supernatant to 2-4 times the weight of Eucystis salsa to obtain Eucystis salsa extract.

[0034] Furthermore, during the ultrafiltration process in step 3, the ultrafiltration temperature is controlled below 30°C.

[0035] The invention also discloses a use of the whitening composition in preparing cosmetics or skin care products.

[0036] Beneficial Effects

[0037] Compared with the prior art, the present invention has at least the following advantages:

[0038] (1) The present invention discloses a whitening composition, which uses white mulberry as the main whitening component, supplemented by Euonymus japonicus, decarboxylated carnosine hydrochloride, and large-flowered cactus flower extract to achieve moisturizing, anti-inflammatory, anti-aging, and skin tightening effects, while comprehensively regulating the skin condition and synergistically improving the skin whitening effect;

[0039] (2) The whitening composition of the present invention is based on the extract of Prunus mume, the extract of Opuntia grandiflora, the extract of Euonymus japonicus and decarboxylated carnosine hydrochloride, and is combined with cyclodextrin to fully inhibit the activity of tyrosinase and inhibit the melanin of B16 cells, thereby synergistically improving the whitening effect of the composition;

[0040] (3) The present invention provides a method for preparing a whitening composition, which maximizes the synergistic whitening effect of the whitening composition through a specific extraction process. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] The present invention is further described below in conjunction with the accompanying drawings and embodiments;

[0042] Figure 1is a picture of a chicken embryo before using the whitening composition of Example 1 of the present invention;

[0043] Figure 2 This is a picture of a chicken embryo after using the whitening composition of Example 1 of the present invention;

[0044] Figure 3 It is a screenshot of the conclusion of the chicken embryo chorioallantoic membrane test report of Example 1 of the present invention.

[0045] Figure 4 It is a screenshot of the conclusion of the tyrosinase inhibition rate detection report of Example 1 of the present invention.

[0046] Figure 5 This is a screenshot of the conclusion of the B16 cell melanin synthesis inhibition rate test report of Example 1 of the present invention. DETAILED DESCRIPTION

[0047] The present invention will be further described below in conjunction with the embodiments, but it does not constitute any limitation to the present invention. Any limited number of modifications made within the scope of the claims of the present invention are still within the scope of the claims of the present invention.

[0048] In order to explain the technical content of the present invention in detail, further description is given below in conjunction with the implementation modes.

[0049] In the following comparative examples, the Bonat Spirulina extract was purchased from New Daily Chemical Trade Development (Hong Kong) Co., Ltd., the brown algae extract was purchased from Shaanxi Tianshengyuan Biotechnology Co., Ltd., the palmitoyl tetrapeptide-7 was purchased from Guangzhou Hemiao Biotechnology Co., Ltd., the purslane extract was purchased from Guangzhou Hemiao Biotechnology Co., Ltd., the seabuckthorn fruit extract was purchased from Zhencui (Guangdong) Innovation Technology Co., Ltd., the nonapeptide-1 was purchased from Guangzhou Hemiao Biotechnology Co., Ltd., and the decarboxylated carnosine hydrochloride was purchased from Guangzhou Hemiao Biotechnology Co., Ltd.

[0050] In the following examples and comparative examples, the Bletilla striata extract was prepared by the following steps:

[0051] Step a: crush the white mulberry into powder;

[0052] Step b: adding 70% ethanol solution with a mass concentration of 15 times the weight of the powder, stirring and extracting at a temperature of 50° C. for 3 hours, and then centrifuging and filtering to obtain a filtrate;

[0053] Step c: Concentrate the filtrate to 3 times the weight of the concentrated juice powder to obtain the iris extract.

[0054] In the following examples and comparative examples, the cactus flower extract was extracted by the following steps:

[0055] Step d: putting the large-flowered cactus flowers into a homogenizer, adding water twice the weight of the large-flowered cactus flowers for homogenization, and filtering to obtain a filtrate;

[0056] Step e: adding water 10 times the weight of the large-flowered cactus flower to the filtrate, stirring and extracting at 70° C. for 3 hours, and then centrifuging to obtain a supernatant;

[0057] Step f: Concentrating the supernatant to twice the weight of the large-flowered cactus flower to obtain the large-flowered cactus flower extract.

[0058] In the following examples and comparative examples, the Eucystis extract was obtained by extraction through the following steps:

[0059] Step g: breaking the cell wall of Eucystis spp. to obtain a broken cell wall product;

[0060] Step h: adding water 10 times the weight of Eucystis to the broken cell wall material, inactivating the enzyme at 70° C. for 40 min, cooling to 50° C. and stirring for extraction for 3 h, and then centrifuging to obtain a supernatant;

[0061] Step i: Concentrate the supernatant to twice the weight of Eucystis salsa to obtain Eucystis salsa extract.

[0062] In the following examples and comparative examples, unless otherwise specified, the parts are parts by weight and the % are percentages by weight.

[0063] The following examples and comparative examples were prepared by the following steps:

[0064] Stir and dissolve the iris extract (or purslane extract, seabuckthorn fruit extract), the large-flowered cactus flower extract, the Euonymus extract (or the spirulina extract, the brown algae extract), decarboxylated carnosine hydrochloride (or palmitoyl tetrapeptide-7, nonapeptide-1), cyclodextrin and water at a temperature of 40° C. for 30 minutes to obtain a mixed solution;

[0065] Step 3: ultrafiltration the mixed solution at an ultrafiltration temperature below 30° C., and sterilization is performed after the ultrafiltration is completed to obtain the whitening composition.

[0066] Example 1

[0067] A whitening composition comprises 15% of iris oleifera extract, 1% of large-flowered cactus flower extract, 1% of Euonymus japonicus extract, 0.3% of decarboxylated carnosine hydrochloride, 0.5% of cyclodextrin, and the balance is water.

[0068] Example 2

[0069] A whitening composition comprises 10% of iris oleifera extract, 1.5% of opuntia grandiflora flower extract, 1.5% of euonymus extract, 0.1% of decarboxylated carnosine hydrochloride, 0.6% of cyclodextrin, and the balance is water.

[0070] Example 3

[0071] A whitening composition comprises 10% of iris oleifera extract, 0.5% of large-flowered cactus flower extract, 0.5% of Eucystis schrenckii extract, 0.5% of decarboxylated carnosine hydrochloride, 0.6% of cyclodextrin, and the balance is water.

[0072] Example 4

[0073] A whitening composition comprises 20% of iris oleifera extract, 1.5% of opuntia grandiflora flower extract, 0.5% of euonymus extract, 0.1% of decarboxylated carnosine hydrochloride, 0.4% of cyclodextrin, and the balance is water.

[0074] Example 5

[0075] A whitening composition comprises 17.5% of a hollyhock extract, 0.75% of a large-flowered cactus flower extract, 0.8% of a euonymus extract, 0.2% of decarboxylated carnosine hydrochloride, 0.55% of cyclodextrin, and the balance is water.

[0076] Example 6

[0077] A whitening composition comprises 13.5% of a hollyhock extract, 1.25% of a large-flowered cactus flower extract, 1.2% of a euonymus extract, 0.4% of decarboxylated carnosine hydrochloride, 0.45% of cyclodextrin, and the balance is water.

[0078] Example 7

[0079] A whitening composition comprises 25% of iris oleifera extract, 5% of cactus flower extract, 3% of Eucystis schrenckii extract, 2% of decarboxylated carnosine hydrochloride, 5% of cyclodextrin, and the balance is water.

[0080] Example 8

[0081] A whitening composition comprises 15% of iris oleifera extract, 10% of opuntia grandiflora flower extract, 10% of euonymus extract, 1% of decarboxylated carnosine hydrochloride, 1% of cyclodextrin, and the balance is water.

[0082] Comparative Example 1

[0083] A whitening composition comprises 15% of a hollyhock extract, 1% of a large-flowered cactus flower extract, 0.3% of decarboxylated carnosine hydrochloride, 0.5% of cyclodextrin, and the balance is water.

[0084] Comparative Example 2

[0085] A whitening composition comprises 15% of a hollyhock extract, 1% of a large-flowered cactus flower extract, 1% of a euonymus extract, 0.5% of cyclodextrin, and the balance is water.

[0086] Comparative Example 3

[0087] A whitening composition comprises 1% of a large-flowered cactus flower extract, 1% of a Eucystis extract, 0.3% of decarboxylated carnosine hydrochloride, 0.5% of cyclodextrin, and the balance is water.

[0088] Comparative Example 4

[0089] A whitening composition comprises 15% of a smilax glabra extract, 1% of a euonymus extract, 0.3% of decarboxylated carnosine hydrochloride, 0.5% of cyclodextrin, and the balance is water.

[0090] Comparative Example 5

[0091] A whitening composition comprises 15% of iris leucophylla extract, 1% of large-flowered cactus flower extract, 1% of spirulina bonaparte extract, 0.3% of decarboxylated carnosine hydrochloride, 0.5% of cyclodextrin, and the balance is water.

[0092] Comparative Example 6

[0093] A whitening composition comprises 15% of iris liliifolia extract, 1% of large-flowered cactus flower extract, 1% of brown algae extract, 0.3% of decarboxylated carnosine hydrochloride, 0.5% of cyclodextrin, and the balance is water.

[0094] Comparative Example 7

[0095] A whitening composition comprises 15% of iris leucophylla extract, 1% of opuntia grandiflora flower extract, 1% of euonymus extract, 0.3% of palmitoyl tetrapeptide-7, 0.5% of cyclodextrin, and the balance is water.

[0096] Comparative Example 8

[0097] A whitening composition comprises 15% of iris leucophylla extract, 1% of opuntia grandiflora flower extract, 1% of euonymus extract, 0.3% of nonapeptide-1, 0.5% of cyclodextrin, and the balance is water.

[0098] Comparative Example 9

[0099] A whitening composition comprises 15% of purslane extract, 1% of large-flowered cactus flower extract, 1% of Eucystis ulmoides extract, 0.3% of decarboxylated carnosine hydrochloride, 0.5% of cyclodextrin, and the balance is water.

[0100] Comparative Example 10

[0101] A whitening composition comprises 15% of seabuckthorn fruit extract, 1% of large-flowered cactus flower extract, 1% of Eucystis ulmoides extract, 0.3% of decarboxylated carnosine hydrochloride, 0.5% of cyclodextrin, and the balance is water.

[0102] Performance Testing

[0103] 1. Security testing

[0104] Test method: Chicken embryo chorioallantoic membrane test

[0105] 1. Purpose and principle of the test

[0106] The chicken embryo chorioallantoic membrane test is an early in vitro eye irritation assessment method. The chorioallantoic membrane (CAM) is a respiratory membrane that surrounds the chicken embryo. This test uses the characteristics of the intact, clear and transparent vascular system of the chorioallantoic membrane of the mid-stage incubated chicken embryo. A certain amount of the test substance is directly exposed to the chicken embryo chorioallantoic membrane. After a period of action, the changes in the toxic effect indicators of the chorioallantoic membrane (such as bleeding, coagulation and vascular melting) are observed. These indicators reflect the changes in the morphological structure, color and permeability of the blood vessels and vascular networks, as well as the phenomena such as the denaturation of the chorioallantoic membrane proteins and the degree of damage. Then, a score is obtained by combining them to evaluate the eye irritation of the test substance.

[0107] The purpose of this test is to test the ability of the test substance to cause toxic changes in the chicken embryo chorioallantoic membrane and to evaluate the elements and processes of the potential eye irritation of the evaluated substance.

[0108] 2. Experimental Group Treatment and Reagents

[0109] Sample group: The whitening compositions prepared in Examples 1 to 8 and Comparative Examples 1 to 10 were diluted into 2% aqueous solutions.

[0110] Negative control: 0.9% saline.

[0111] Positive control: 1% sodium dodecyl sulfate (SDS) solution.

[0112] 3. Experimental steps

[0113] In this test, 6 chicken embryos were selected for each group, and the conditions of the chorioallantoic membrane were recorded with a camera. A polytetrafluoroethylene resin ring was placed on the chorioallantoic membrane of the chicken embryo and photographed. The sample to be tested was added to the polytetrafluoroethylene resin ring, the time of adding the sample was recorded, and the air chamber was covered with a moistened plastic wrap. The chicken embryo was moved to a constant temperature and humidity chamber for culture, and the degree of change of each toxic effect was observed.

[0114] 4. Calculation of results

[0115] The endpoint evaluation method was used to perform the test and the endpoint evaluation (ES) was calculated with two decimal places. Each chicken embryo was scored as the sum of the bleeding, coagulation and vascular melting observed in each chicken embryo. The average value of the mathematical sum obtained for ES-6 chicken embryos was taken.

[0116] The average ES score was calculated using the following formula.

[0117]

[0118] 5. Result judgment criteria

[0119] ES≤4, non-irritating;

[0120] 4<ES≤12, mild irritation;

[0121] 12<ES<16, moderate irritation;

[0122] ES ≥ 16, strong irritant / corrosive.

[0123] 2. Test Results

[0124] According to the above test method, samples of Examples 1-8 and Comparative Examples 1-10 were tested, and the ES values ​​were all less than 4, and the results were all judged to be non-irritating.

[0125] Among them, the results of chicken embryos before and after use in Example 1 are as follows Figure 1-2 As shown, the sample test report conclusion screenshot of Example 1 is shown in Figure 3 ;

[0126] This proves that the technical solution of the present invention is mild and non-irritating.

[0127] 2. Tyrosinase inhibition rate test

[0128] Test method:

[0129] 1. Experimental Group Treatment and Reagents

[0130] Sample group: The whitening compositions prepared in Examples 1 to 8 and Comparative Examples 1 to 10 were diluted into 2% aqueous solutions respectively.

[0131] Positive control: 1 mg / mL, 0.2 mg / mL, 0.04 mg / mL, 0.008 mg / mL kojic acid solution (using pH 6.8 disodium hydrogen phosphate-citrate buffer).

[0132] Negative control: pure water.

[0133] Reagents: Tyrosinase solution: prepared with pH 6.8 disodium hydrogen phosphate-citric acid buffer, 100u / mL; L-dopa solution: prepared with pH 6.8 disodium hydrogen phosphate-citric acid buffer, 1mg / mL.

[0134] 2. Experimental steps

[0135] The test was conducted according to the standard T / SHRH 015-2018 "Cosmetics - Tyrosinase Activity Inhibition Test Method". Referring to Table 1, 10mL test tubes were used to set up sample tubes (T), sample background (T0), enzyme reaction tubes (C) and solvent background (C0). Three parallel tubes were required for each sample tube (T) of each tested concentration, and three parallel tubes were also required for the enzyme reaction tube (C).

[0136] Table 1 Sample addition requirements

[0137]

[0138] Add 1 mL of sample solution of the same concentration to the sample tube (T) and sample background (T0), and add 1 mL of disodium hydrogen phosphate-citric acid buffer to the enzyme reaction tube (C) and solvent background (C0). Add 0.5 mL of tyrosinase solution to the sample tube (T) and enzyme reaction tube (C), replace the sample background (T0) and solvent background (C0) with 0.5 mL of disodium hydrogen phosphate-citric acid buffer, mix the sample and tyrosinase thoroughly, and incubate in a 37°C water bath for 10 minutes. Add 2 mL of L-DOPA solution to each tube in turn, control the reaction time of each tube to 5 minutes, and immediately transfer the reaction solution of each tube into a cuvette, and measure the absorbance at 475 nm.

[0139] 3. Calculation of results

[0140] The tyrosinase inhibition rate was calculated according to the following formula.

[0141]

[0142] In the formula

[0143] T—is the absorbance of the sample tube, i.e., the absorbance of the solution after the sample reacts with tyrosinase;

[0144] T0—is the background absorbance value of the sample;

[0145] C—is the average of three absorbance values ​​of the enzyme reaction tube, that is, the absorbance value of the tyrosinase and DOPA reaction when no sample is added;

[0146] C0—is the background absorbance of the solvent.

[0147] 4. Result judgment criteria

[0148] The positive control tyrosinase inhibition rate is greater than 50%, and the reaction system is effective. When the sample tyrosinase inhibition rate is significantly different from the negative control (P < 0.05), it can be considered that the test sample has a certain whitening effect.

[0149] The whitening compositions obtained in Examples 1-8 and Comparative Examples 1-10 were tested according to the above test method, and the results are shown in Table 2;

[0150] Table 2 Tyrosinase inhibition test results of whitening compositions of Examples 1-8 and Comparative Examples 1-10

[0151] Group Tyrosinase inhibition rate (%) P-value Example 1 81.14 ＜0.050 Example 2 52.47 ＜0.050 Example 3 60.73 ＜0.050 Example 4 78.82 ＜0.050 Example 5 67.62 ＜0.050 Example 6 54.53 ＜0.050 Example 7 88.77 ＜0.050 Example 8 84.56 ＜0.050 Comparative Example 1 38.58 ＜0.050 Comparative Example 2 29.36 ＜0.050 Comparative Example 3 6.75 ＜0.050 Comparative Example 4 43.18 ＜0.050 Comparative Example 5 39.23 ＜0.050 Comparative Example 6 43.69 ＜0.050 Comparative Example 7 35.22 ＜0.050 Comparative Example 8 33.73 ＜0.050 Comparative Example 9 44.31 ＜0.050 Comparative Example 10 50.67 ＜0.050 Negative control -2.18 / Positive Control 94.48 ＜0.050

[0152] According to the results in Table 2:

[0153] According to the data of Examples 1-8, the compound of the present invention using the extract of Psoralea corylifolia, the extract of Opuntia grandiflora, the extract of Euonymus japonicus and decarboxylated carnosine hydrochloride can fully inhibit the activity of tyrosinase;

[0154] According to the data comparison of Example 1 and Comparative Examples 1-4, in the absence of any one of the white osmanthus extract, the large-flowered cactus flower extract, the eucystis extract and the decarboxylated carnosine hydrochloride, the tyrosinase activity inhibition rate shown in Comparative Examples 1-4 will be greatly reduced, indicating that the synergistic effect of improving the tyrosinase activity inhibition effect of the present invention is based on the white osmanthus extract, the large-flowered cactus flower extract, the eucystis extract and the decarboxylated carnosine hydrochloride.

[0155] However, since Comparative Example 3 lacks the whitening ingredient, the Bletilla striata extract, it does not show any obvious effect of inhibiting tyrosinase activity. The only single-digit tyrosinase inhibition rate may be due to the fact that some components of the Euonymus japonicus extract or the Opuntia grandiflora flower extract have a certain effect of inhibiting tyrosinase activity. This further proves that the addition of the Opuntia grandiflora flower extract, Euonymus japonicus extract and decarboxylated carnosine hydrochloride can synergistically improve the whitening effect of the Bletilla striata extract.

[0156] According to the data comparison of Example 1 and Comparative Examples 5 and 6, it can be seen that the use of other algae extracts to replace the Eucystis extract cannot produce a synergistic effect of improving the tyrosinase inhibition rate of the composition with the white lily extract, the large-flowered cactus flower extract and decarboxylated carnosine hydrochloride; and among them, Comparative Example 6 uses a brown algae extract of the same family and similar efficacy as the Eucystis extract. It is found that although the tyrosinase inhibition rate is improved to a certain extent compared with the technical scheme of Comparative Example 5, the improvement is significantly reduced compared with the technical scheme of Example 1, indicating that the use of the Eucystis extract, the white lily extract, the large-flowered cactus flower extract and the decarboxylated carnosine hydrochloride in the present invention can produce a significant effect of improving the tyrosinase inhibition rate.

[0157] According to the data comparison between Example 1 and Comparative Examples 7 and 8, it can be seen that the use of other peptides instead of decarboxylated carnosine hydrochloride cannot produce a synergistic effect of improving the tyrosinase inhibition rate of the composition with the Bletilla striata extract, the Opuntia grandiflora flower extract and the Euonymus japonicus extract.

[0158] According to the data comparison between Example 1 and Comparative Examples 9 and 10, it can be seen that the use of other whitening functional ingredients such as the purslane extract of Comparative Example 9 and the sea buckthorn fruit extract of Comparative Example 10 cannot produce a synergistic effect of improving the tyrosinase inhibition rate of the composition with the large-flowered cactus flower extract, the Eucystis extract and the decarboxylated carnosine hydrochloride, indicating that the large-flowered cactus flower extract, the Eucystis extract and the decarboxylated carnosine hydrochloride of the present invention improve the whitening effect of the white lily extract through the synergistic relationship between the functional ingredients, and do not have the effect of improving the tyrosinase inhibition rate of all whitening functional ingredients.

[0159] 3. B16 cell melanin synthesis inhibition rate test

[0160] Test method:

[0161] 1. Purpose and principle of the test

[0162] B16 mouse melanoma cells are a commonly used cell model for studying melanogenesis. The whitening effect of cosmetics is evaluated by measuring the changes in melanin synthesis after whitening ingredients act on melanocytes.

[0163] This test refers to the laboratory method and compares the results of the cell melanin synthesis inhibition rate determination of the test sample with that of the negative control. If the test sample can detect the effect of inhibiting B16 cell melanin synthesis and there is a significant difference compared with the negative control (P<0.05), it can be considered that the test sample has a certain whitening effect.

[0164] 2. Test indicators

[0165] Melanin synthesis inhibition rate, judgment standard: If the test sample can detect the effect of inhibiting B16 cell synthesis of melanin, and there is a significant difference with the negative control, it reflects that the sample has a certain whitening effect.

[0166] 3. Experimental Materials and Methods

[0167] 3.1 Instruments and Equipment

[0168] RT-6100 ELISA Analyzer

[0169] 3.2 Cells used in the experiment

[0170] B16 mouse melanoma cells

[0171] 3.3 Test methods

[0172] 3.3.1 Treatment of test samples: The whitening compositions of Examples 1-8 and Comparative Examples 1-10 were diluted with pure water to a solution with a volume fraction of 4%, and then filtered with a 0.22 μm filter, and the filtrate was collected as the sample mother solution.

[0173] Negative control: basal medium.

[0174] 3.3.2 Test operation steps

[0175] Cell activity test: B16 cells were plated in 96-well plates. After 24 hours, the culture medium was discarded and basal culture medium containing test samples at different concentrations was added. After 24 hours, OD490nm was detected by MTT method and the effect of test samples on B16 cell activity was analyzed by t-test.

[0176] Determination of the inhibition rate of melanin synthesis in B16 cells: B16 cells were plated on a 6-well plate, cultured for 24 hours, and then replaced with a basal medium containing different concentrations of test samples. After two changes of medium, the cells were washed twice with PBS, 200 μL of 0.25% trypsin was added to each well to digest the cells, and the cells were collected into a centrifuge tube and centrifuged for 5 minutes. 200 μL of melanin extract was added to each tube, shaken evenly, and transferred to a 96-well plate. The absorbance of each well at 405 nm was detected by an ELISA instrument.

[0177] 4. Calculation formula

[0178]

[0179] Where: T—absorbance of test sample hole;

[0180] C—average of three absorbance values ​​of the negative control group;

[0181] C0—Background absorbance of melanin extract.

[0182] 5. Data Analysis

[0183] The statistical analysis software was SPSS, and the comparison between the test sample and the negative control was performed using the independent sample t test. The above statistical analyses were all two-tailed tests, and the significance level was α = 0.05. P>0.05 indicated that there was no significant difference between the two groups; P<0.05 indicated that there was a significant difference between the two groups.

[0184] The whitening compositions obtained in Examples 1-8 and Comparative Examples 1-10 were tested according to the above test method. When the sample concentration was 0.25%, the results were shown in Table 3.

[0185] Table 3 Test results of the inhibition rate of melanin synthesis in B16 cells of the whitening compositions of Examples 1-8 and Comparative Examples 1-10

[0186] Group B16 cell melanin synthesis inhibition rate (%) P-value Example 1 36.44 ＜0.050 Example 2 30.26 ＜0.050 Example 3 34.65 ＜0.050 Example 4 35.48 ＜0.050 Example 5 34.34 ＜0.050 Example 6 32.15 ＜0.050 Example 7 38.12 ＜0.050 Example 8 37.41 ＜0.050 Comparative Example 1 19.30 ＜0.050 Comparative Example 2 12.85 ＜0.050 Comparative Example 3 0.64 ＜0.050 Comparative Example 4 26.21 ＜0.050 Comparative Example 5 24.73 ＜0.050 Comparative Example 6 26.27 ＜0.050 Comparative Example 7 23.84 ＜0.050 Comparative Example 8 22.48 ＜0.050 Comparative Example 9 27.42 ＜0.050 Comparative Example 10 28.08 ＜0.050 Positive Control 49.97 ＜0.050

[0187] According to the results in Table 3:

[0188] According to the data of Examples 1-8, the compound of the present invention using the extract of Psoralea corylifolia, the extract of Opuntia grandiflora, the extract of Streptocarpus oleraceus and decarboxylated carnosine hydrochloride can fully inhibit the synthesis of melanin in B16 cells;

[0189] According to the data comparison of Example 1 and Comparative Examples 1-4, in the absence of any one of the white osmanthus extract, the large-flowered cactus flower extract, the eucystis extract and the decarboxylated carnosine hydrochloride, the B16 cell melanin synthesis inhibition rate shown in Comparative Examples 1-4 will be greatly reduced, indicating that the synergistic effect of improving the inhibition of B16 cell melanin synthesis of the present invention is based on the white osmanthus extract, the large-flowered cactus flower extract, the eucystis extract and the decarboxylated carnosine hydrochloride.

[0190] However, since Comparative Example 3 lacks the whitening ingredient, the Bletilla striata extract, it does not show any obvious effect of inhibiting B16 cell melanin synthesis. The only single-digit inhibition rate of B16 cell melanin synthesis may be due to the fact that some components of the Bletilla striata extract or the Opuntia grandiflora flower extract have a certain effect of inhibiting B16 cell melanin synthesis. This further proves that the addition of the Bletilla striata extract, the Bletilla striata extract and the decarboxylated carnosine hydrochloride can synergistically improve the whitening effect of the Bletilla striata extract.

[0191] According to the data comparison of Example 1 and Comparative Examples 5 and 6, it can be seen that the use of other algae extracts to replace the Eucystis extract cannot produce a synergistic effect with the white lily extract, the large-flowered cactus flower extract and decarboxylated carnosine hydrochloride to improve the B16 cell melanin synthesis inhibition rate of the composition; and Comparative Example 6 uses a brown algae extract of the same genus and similar efficacy as the Eucystis extract. It is found that although the B16 cell melanin synthesis inhibition rate is improved to a certain extent compared with the technical scheme of Comparative Example 5, the improvement is significantly reduced compared with the technical scheme of Example 1, indicating that the use of the Eucystis extract, the white lily extract, the large-flowered cactus flower extract and the decarboxylated carnosine hydrochloride in the present invention can produce a significant effect of improving the B16 cell melanin synthesis inhibition rate.

[0192] According to the data comparison between Example 1 and Comparative Examples 7 and 8, it can be seen that the use of other peptides instead of decarboxylated carnosine hydrochloride cannot produce a synergistic effect of improving the B16 cell melanin synthesis inhibition rate of the composition with the Bletilla striata extract, the large-flowered cactus flower extract and the Euonymus japonicus extract.

[0193] According to the data comparison between Example 1 and Comparative Examples 9 and 10, it can be seen that the use of other whitening functional ingredients such as the Portulaca oleracea extract of Comparative Example 9 and the Hippophae rhamnoides fruit extract of Comparative Example 10 cannot produce a synergistic effect of improving the B16 cell melanin synthesis inhibition rate of the composition with the Opuntia grandiflora flower extract, the Eucystis ulmoides extract and the decarboxylated carnosine hydrochloride, indicating that the Opuntia grandiflora flower extract, the Eucystis ulmoides extract and the decarboxylated carnosine hydrochloride of the present invention improve the whitening effect of the Brussels sprout extract through the synergistic relationship between the functional ingredients, and do not have the effect of improving the B16 cell melanin synthesis inhibition rate of all whitening functional ingredients.

[0194] Application Examples

[0195] A whitening composition essence liquid consists of phase A, phase B, phase C and phase D.

[0196] Table 4 A whitening composition essence raw material ratio

[0197]

[0198] A whitening composition essence is prepared by the following steps: phase B is completely dissolved in advance, phase C is completely heated in advance and dissolved, phase A is added to an emulsifying pot, heated to 85°C, stirred and homogenized, the pre-dissolved phase B is added, then the temperature is lowered to 45°C, the pre-dissolved phases C and D are added, and stirred at 30r / min for 10min to obtain the whitening composition essence.

[0199] The embodiments presented herein are only embodiments selected according to the combination of all possible embodiments. The appended claims should not be limited by the embodiments describing the present invention. Some numerical ranges used in the claims include sub-ranges therein, and the changes in these ranges should also be covered by the appended claims.

Claims

1. A whitening composition, characterized in that: The composition comprises, by weight percentage, 8-25% of a Bletilla striata extract, 0.1-10% of a large-flowered cactus flower extract, 0.1-10% of a Eucystis fasciata extract, 0.1-5% of a decarboxylated carnosine hydrochloride, 0.1-5% of a cyclodextrin, and the balance of water; The white osmanthus extract is obtained by extracting through the following steps: Step a: crush the white mulberry into powder; Step b: adding 60-80% ethanol solution with a mass concentration of 10-20 times the weight of the powder, stirring and extracting at a temperature of 48-52°C for 2-4h, and then centrifuging and filtering to obtain a filtrate; Step c: Concentrating the filtrate to a weight of 2-4 times the weight of the concentrated juice powder to obtain a iris extract; The large-flowered cactus flower extract is obtained by extracting through the following steps: Step d: putting the large-flowered cactus flowers into a homogenizer, adding water twice the weight of the large-flowered cactus flowers for homogenization, and filtering to obtain a filtrate; Step e: adding water 10-14 times the weight of the large-flowered cactus flower to the filtrate, stirring and extracting at a temperature of 65-75° C. for 2-4 hours, and then centrifuging to obtain a supernatant; Step f: concentrating the supernatant to 2-4 times the weight of the large-flowered cactus flower to obtain a large-flowered cactus flower extract; The Eucystis extract is obtained by extracting through the following steps: Step g: breaking the cell wall of Eucystis spp. to obtain a broken cell wall product; Step h: adding water 10-14 times the weight of Eucystis to the broken cell wall material, inactivating the enzyme at 65-75°C for 30-60 minutes, cooling to 45-55°C, stirring and extracting for 2-4 hours, and then centrifuging to obtain a supernatant; Step i: Concentrate the supernatant to 2-4 times the weight of Eucystis salsa to obtain Eucystis salsa extract.

2. A method for preparing the whitening composition according to claim 1, characterized in that: The steps include: Step 1: extracting the white osmanthus, the flower of Opuntia grandiflora, and the Streptospira oleracea to obtain the white osmanthus extract, the flower of Opuntia grandiflora, and the Streptospira oleracea extract; Step 2: stirring and dissolving the white mulberry extract, the large-flowered cactus flower extract, the Eucystis extract, decarboxylated carnosine hydrochloride, cyclodextrin and water at a temperature of 40-50° C. for 20-30 minutes to obtain a mixed solution; Step 3: The mixed solution is subjected to ultrafiltration and sterilization treatment in sequence to obtain the whitening composition.

3. The method for preparing the whitening composition according to claim 2, characterized in that: During the ultrafiltration treatment in step 3, the ultrafiltration temperature is controlled below 30°C.

4. Use of the whitening composition according to claim 1 in preparing cosmetics or skin care products.

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