A composition with a stable white function and a preparation method and application thereof

Abstract

This invention provides a composition with whitening function, its preparation method, and its application, belonging to the field of cosmetic technology. The composition comprises black tea extract and tranexamic acid in a mass ratio of 1:0.5-55. The two components in this composition have a synergistic effect, achieving enhanced whitening and inhibition of post-inflammatory hyperpigmentation by acting on multiple target sites, thus achieving a whitening effect. Both components are mild in nature, the preparation method is simple and convenient, and it has good practical value.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, specifically to a composition with whitening function, its preparation method, and its application. Background Technology

[0002] Dull skin is a common skin problem. Considering the pursuit of skin whitening by many women, the development of whitening products and raw materials is one of the most popular research directions in the cosmetics market, holding a significant position. Human skin's epidermis, dermis, and subcutaneous tissue all contain pigments, and the synthesis and deposition of these pigments affect skin color. Among them, melanin deposition in keratinocytes is the most significant factor leading to dull skin. Currently, whitening cosmetic raw materials mainly focus on inhibiting key processes of melanin production in melanocytes, such as inhibiting tyrosinase activity, inhibiting melanin synthesis signaling pathways, and interfering with melanosome transfer. Although some developed whitening raw materials, such as ascorbic acid and its derivatives, nicotinic acid derivatives, arbutin and its derivatives, and glycyrrhizin, have good whitening effects by inhibiting key processes of melanin production, raw materials that possess both whitening and whitening-stabilizing properties are still relatively scarce. Post-inflammatory hyperpigmentation, caused by excessive melanin secretion following skin inflammation, is a significant factor leading to skin darkening. Since photosensitive hyperplasia (PIH) is exacerbated by photodeposition, it primarily occurs on the face, negatively impacting patients' appearance and psychological well-being, thus reducing their quality of life. Therefore, developing cosmetic ingredients that can both whiten and inhibit PIH to achieve a "whitening-stabilizing" effect is of great significance.

[0003] Dark tea is a post-fermented tea obtained through microbial fermentation. It contains a variety of functional components, including catechins and their derivatives, flavonols, phenolic acids, alkaloids, amino acids, terpenes, polysaccharides, and vitamins. These functional components originate from compounds formed during the natural growth and metabolism of the tea plant and metabolic products produced by the microbial community during dark tea fermentation. Due to its unique processing techniques, excellent quality, and outstanding health benefits, dark tea is widely popular and used in the food industry. However, relatively little research has been conducted on its efficacy in cosmetics.

[0004] Tranexamic acid (also known as aminomethyl tranexamic acid or tranexamic acid) is a mild and stable skin-whitening ingredient. It can inhibit the production and deposition of melanin by competitively inhibiting the binding of plasminogen to keratinocytes or melanocytes. The activation of plasmin not only stimulates the release of inflammatory factors (such as prostaglandin PGE2, interleukin IL-β, IL-6, etc.), thereby activating tyrosinase (a key enzyme in melanin synthesis) and leading to melanin production, but also promotes the release of melanocyte-stimulating hormone (α-MSH) from keratinocytes, which then binds to the melanocortin-1 receptor (MC1R receptor) on melanocytes, activating tyrosinase.

[0005] Invention patent CN111544309B discloses a freeze-dried solid facial mask containing bioactive ingredients and its preparation method. The freeze-dried solid facial mask containing bioactive ingredients is prepared by drying a facial mask cloth impregnated with a mask liquid at low temperature. The mask liquid includes phase A, phase B, and phase C. Phase A includes water, glycerin, propylene glycol, plant extracts, sodium hyaluronate, tranexamic acid, raspberry ketone glucoside, sclerotium gum, erythritol, allantoin, thickener, and p-hydroxyacetophenone. Phase B includes olive fruit oil, grape seed extract, jojoba seed extract, sweet orange essential oil, vitamin E, dioctyl carbonate, and hydrogenated lecithin. Phase C is adipose-derived mesenchymal stem cell lysate.

[0006] There is a lack of research in the existing technology on the preparation of whitening-stabilizing compositions by combining black tea extract and tranexamic acid. In order to fill the gap in this field and study the interaction between the two, this invention proposes a composition containing black tea extract and tranexamic acid with whitening function, as well as its preparation method and application. Summary of the Invention

[0007] This invention discovers that the combination of black tea extract and tranexamic acid has a good effect on inhibiting melanin production and reducing PIH. Through synergistic effect, it inhibits inflammatory factors and melanin production, thus achieving the effect of stabilizing white skin.

[0008] This invention discloses a composition with whitening effects, the components of which are black tea extract and tranexamic acid. This composition with whitening effects may be named "StaWhite-T Whitening Factor" in this invention.

[0009] In one embodiment, the mass ratio of black tea extract to tranexamic acid in a composition having whitening effects is 1:0.5-55.

[0010] In one embodiment, the mass ratio of black tea extract to tranexamic acid in a composition with whitening effect is 1:0.5-25, 1:0.8-20, 1:1-15, 1:2-10, etc.

[0011] In one embodiment, the method for preparing the black tea extract is as follows: mixing black tea with a polyol solution, heating and extracting, and then filtering.

[0012] In one embodiment, the polyol includes at least one of ethylene glycol, propylene glycol, and butanediol.

[0013] In one embodiment, the polyol is butanediol.

[0014] In one embodiment, the polyol solution in the preparation method of black tea extract is composed of water and polyol in a volume ratio of (5-7):(2-3.5).

[0015] In one embodiment, the polyol solution in the preparation method of black tea extract is composed of water and polyol in a volume ratio of 7:3.

[0016] In one embodiment, in the method for preparing black tea extract, the heating and extraction temperature and time are 50-60℃ and 30-200 min.

[0017] In one embodiment, the preparation method of black tea extract includes: mixing black tea with a polyol solution (composed of deionized water and polyol in a volume ratio of (5-7):(2-3.5)) at a mass ratio of 1:(3-8), heating and extracting in a water bath at 50-60℃ for 30-120 min to obtain the material, then filtering with a filter screen to obtain a coarse filtrate, and then filtering with a filter membrane to obtain the filtrate, which is the black tea extract.

[0018] The present invention also provides a method for preparing a composition with whitening effect, comprising the steps of mixing raw materials and packaging.

[0019] The present invention also provides the application of a composition with whitening effect in the preparation of cosmetics, wherein the composition with whitening effect has a weight percentage of 1.5-8.0% in the cosmetics.

[0020] In one embodiment, the cosmetic also includes 2-10% Cornus officinalis fruit extract. The mass ratio of the Cornus officinalis fruit extract to the composition having whitening effects in the cosmetic is 1:0.6-0.85.

[0021] In one embodiment, the preparation method of the Cornus officinalis fruit extract includes: pulping Cornus officinalis fruit with water, sterilizing, adding sucrose and Bifidobacterium for fermentation, sterilizing, first concentration, then using 20-45wt% ethanol solution as extraction solvent for heating and extraction, and then filtering to obtain the filtrate, which is the Cornus officinalis fruit extract.

[0022] In one embodiment, during the preparation of the Cornus officinalis fruit extract, the weight of water added during pulping is 4-15 times the weight of the Cornus officinalis fruit. The fermentation process using Bifidobacterium includes: adding sucrose and then adding Bifidobacterium for fermentation. The amount of Bifidobacterium added is 10... 8 -10 11 One cell / g slurry. Fermentation temperature and time: 25-40℃, 50-120 hours. First concentration to 1-1.3 times the weight of Cornus officinalis fruit. Heating extraction temperature and time: 40-60℃, 5-10 hours.

[0023] The beneficial effects of this invention are:

[0024] The composition of the present invention also has a whitening effect: the black tea extract therein can inhibit key targets in the melanin production process, such as melanocortin-1 receptor (MC1R), microphthalmia-associated transcription factor (MITF), tyrosinase (TYR), tyrosinase-associated protein-1 (TYR-1), and tyrosinase-associated protein-2 (TYR-2), thereby inhibiting melanin production; at the same time, the black tea extract can also inhibit the production of UVB-induced inflammatory factors (IL-1β, IL-6, and PGE2) and paracrine melanin-producing factors, such as α-melanocyte-stimulating hormone (α-MSH), basic fibroblast growth factor (bFGF), and endothelin-1 (ET-1), thereby inhibiting PIH and achieving the whitening effect. Tranexamic acid can inhibit key factors interfering with melanin production—the activation of plasmin in the skin under ultraviolet radiation and the release of inflammatory factors and melanin-producing factors (IL-1β, IL-6, ET-1, arachidonic acid metabolites, and PGE2, etc.), thereby achieving the effect of inhibiting melanin production. The mechanisms of action of black tea extract and tranexamic acid are complementary. Experimental tests and calculations of the combination index using Compusyn software demonstrate that the combination of the two has a synergistic whitening effect.

[0025] Tests on anti-inflammatory and melanin-inhibiting effects show that, in this invention, tranexamic acid and black tea extract exhibit a synergistic effect within a weight ratio of 1:0.5-55, effectively enhancing the whitening effect of cosmetic products. Attached Figure Description

[0026] Figure 1 Effects of 0.5 μg / mg dark tea extract on UVB-induced paracrine melanin-producing factors (α-MSH, bFGF, and ET-1).

[0027] Figure 2 The effect of 0.5 μg / mg black tea extract on key targets for regulating melanin production: MC1R, MITF, TYR, TYR-1, and TYR-2. Detailed Implementation

[0028] The present invention will be further illustrated by the following specific solutions, which are part of the specific content of the present invention.

[0029] Dark tea: Anhua dark tea. Polyol: 1,3-Butanediol.

[0030] 1. Preparation of dark tea extract

[0031] Black tea was mixed with a polyol solution (composed of deionized water and polyol in a volume ratio of 7:3) at a mass ratio of 1:5. The mixture was heated in a water bath at 55°C and stirred at 60 rpm for 100 min to obtain an extract. The extract was then filtered through a 600-mesh filter to obtain a coarse filtrate. This coarse filtrate was then filtered sequentially through 10 μm, 0.8 μm, 0.45 μm, and 0.22 μm filter membranes to obtain the filtrate, which is the black tea extract.

[0032] 2. Effects of dark tea extract on UVB-induced paracrine melanin-producing factors

[0033] HaCat cells were taken at a rate of 2 × 10⁻⁶. 5 Cells were seeded at 0.5 mL DMEM medium per well in 24-well plates, with a density of 1 / mL. Cells were allowed to adhere and confluence to 80%, followed by two washes with PBS. The control group received no radiation treatment and was cultured directly in 0.5 mL DMEM medium for 24 hours. The control and experimental groups were treated with PBS, exposing the cells to 30 mJ / cm². 2 Irradiate the cells under UVB with the lid off. After irradiation, discard the PBS. Add 0.5 mL of DMEM medium to the control group and 0.5 μg / mg DMEM medium containing black tea extract to the experimental group, respectively. Continue culturing for 24 hours. Collect the supernatant and measure the levels of paracrine melanin-producing factors α-MSH, bFGF, and ET-1 secreted by the cells according to the α-MSH, bFGF, and ET-1 kit. Results are shown below. Figure 1 .

[0034] from Figure 1 It can be seen that the secretion of paracrine melanin-producing factors, including α-MSH, bFGF, and ET-1, in HaCaT cells under UVB irradiation was significantly enhanced, indicating that UVB induced the production of paracrine factors in HaCaT cells. These paracrine factors can induce melanin production in melanocytes through different mechanisms. In contrast, the production of paracrine factors in cells incubated with black tea extract was significantly reduced compared to the UVB stimulation group, and the expression levels of α-MSH, bFGF, and ET-1 were significantly decreased.

[0035] 3. The effect of dark tea extract on key targets regulating melanin production

[0036] Take B16 cells at a rate of 2 × 10 5 Cells were seeded at a density of 2.0 mL / well in 6-well plates (DMEM medium + cells). After 24 h of culture, the blank control group and the experimental group were respectively treated with DMEM medium and DMEM medium containing 0.5 μg / mg black tea extract, with 2.0 mL added to each group. Cells were cultured for another 24 h. The DMEM medium was discarded, and 400 μL of trypsin was added to each well for 2 min of digestion. 800 μL of DMEM medium was added to stop the digestion, and the cells were gently pipetted until suspended. The cells were then transferred to centrifuge tubes and centrifuged at 4°C (2000 r / min, 5 min). The supernatant was discarded, and the cells were resuspended in PBS. The cells were centrifuged at 4°C (2000 r / min, 5 min), and the supernatant was discarded. The cell pellet was collected.

[0037] Next, total RNA was extracted from the treated cells using Trizol reagent, and its purity (260 / 280 nm absorbance ratio) was measured and quantified using a nucleic acid and protein analyzer. Subsequently, the RNA was reverse transcribed into cDNA using the PrimeScript RT Master Mix reverse transcription kit. Using the cDNA as a template, quantitative RT-PCR analysis was performed using SYBR Premix Ex Taq premixed buffer in a real-time fluorescence quantitative PCR instrument. The reaction program was set to 40 cycles: 95℃ pre-denaturation for 30 s; each cycle of the amplification phase included 95℃ denaturation for 5 s, followed by 60℃ annealing / extension for 34 s, with fluorescence signals acquired during the extension phase. The primer sequences used for the test are as follows: MC1R, forward primer: 5'-CTCATTGACGTGCTCATCTGTGG-3', reverse primer: 5'-TGCTTGTAGTAGGTGATAAAGAGGGT-3'; MITF, forward primer: 5'-GCCCTATGGCTATGCTCACTCTT-3', reverse primer: 5'-TGTTCATACCTGGGCACTCACTC-3'; TYR, forward primer: 5'-ATCCTAACTTACTCAGCCCAGCA-3', reverse primer: 5'-CTCAGGTGTTCCATCGCATAAA-3'; TYRP-1, forward primer: 5'-TTCGTTGGAGCTGTGATTGTTG-3', reverse primer: 5'-AGGAATAATGTTGAAAGGTGGGG-3'; TYRP-2, Forward primer: 5'-CAGAAATAATGAGAAACTGCCAACC-3', Reverse primer: 5'-TCCGTCTGCTTTATCAAACCCT-3'. Quantitative PCR was performed using fluorescence 2... -△△CtRelative expression levels were calculated using the method (compared to the expression level of the blank control group, which was set to 1.0). The mRNA levels of each target gene were normalized to GAPDH levels, and the results are shown in [Figure number missing]. Figure 2 .

[0038] from Figure 2 It can be seen that dark tea extract significantly reduced the expression levels of key regulators of melanin production, including MC1R, MITF, TYR, TYRP-1, and TYRP-2. This is because dark tea extract can inhibit the secretion of UVB-induced α-MSH, reducing its binding to MC1R, thus inhibiting the key target of melanin production at its source. Subsequently, it also inhibited the expression and activity of MITF, thereby downregulating the expression of its downstream proteins such as TYR, TYRP-1, and TYRP-2 genes, ultimately inhibiting melanin production.

[0039] 4. Anti-inflammatory test

[0040] We tested single components (black tea extract or tranexamic acid) and composite materials (a mixture of black tea extract and tranexamic acid) with different proportions. After dissolving the raw materials in DMEM medium to form a certain content, we used a kit to test the effect of different components on the secretion of inflammatory factor (PGE2) in HaCaT cells under UVB stimulation.

[0041] HaCat cells were processed at a rate of 2 × 10 5 Cells were seeded at 0.5 mL / well in 24-well plates using DMEM medium and water, and allowed to adhere to the plates until 80% confluence. After washing twice with PBS, cells in the control and experimental groups were exposed to PBS at a concentration of 30 mJ / cm². 2 Irradiate the samples under UVB with the lid off. After irradiation, discard the PBS. Add pure DMEM medium to the control group and DMEM medium containing different amounts of a single component (black tea extract or tranexamic acid) or DMEM medium containing different proportions of a combination (black tea extract + tranexamic acid) to the experimental group (the composition and concentration are shown in Table 1). As shown in Table 3, the amount added to each group was 0.5 mL. The content of inflammatory factors secreted in the cell supernatant was measured according to the PGE2 kit procedure (standard curve method), and the PGE2 inhibition rate was obtained. The formula for calculating the PGE2 inhibition rate is as follows:

[0042]

[0043] In formula (1): E1 and E0 represent the mean PGE2 content of the sample group and the blank control group, respectively.

[0044] Based on the inhibitory effect test data of single components and compositions in different proportions, the combination index was calculated using Compusyn software. CI>1 indicates antagonistic effect, CI=1 indicates additive effect, and CI<1 indicates synergistic effect.

[0045] Table 1. PGE2 inhibition rate of black tea extracts with different concentrations

[0046] Content of dark tea extract (μg / mg) PGE2 inhibition rate (%) 0.50 5.1 1.00 20.5 1.50 32.8 2.00 45.7 2.50 57.4 3.00 69.6 3.50 74.7 4.00 81.9 4.50 88.6

[0047] Table 2. PGE2 inhibition rate of different concentrations of tranexamic acid

[0048] Tranexamic acid content (μg / mg) PGE2 inhibition rate (%) 0.50 3.9 1.00 8.8 2.00 15.6 4.00 25.4 8.00 37.9 16.00 56.5 24.00 68.7 30.00 83.4

[0049] Table 3. PGE2 inhibition rate and combination index (CI) of compositions with different proportions

[0050] Content of dark tea extract (μg / mg) Tranexamic acid content (μg / mg) PGE2 inhibition rate (%) CI value 0.50 0.50 11.8 0.952 0.50 1.00 17.3 0.942 0.50 2.00 28.6 0.858 0.50 4.00 49.4 0.681 0.50 8.00 69.1 0.580 0.50 16.00 79.8 0.623 0.50 24.00 83.5 0.708 0.50 30.00 85.9 0.733

[0051] The results showed that both black tea extract and tranexamic acid, as single components, could inhibit the release of the inflammatory factor PGE2 in UVB-induced keratinocytes. The combination index of the compositions with different ratios (1:1~1:60) calculated by Compusyn software was <1, indicating that the compositions have synergistic effects, and that the combination of the two is necessary. Moreover, the ratio of black tea extract to tranexamic acid in the composition in the range of 1:8~1:32 had a more efficient anti-inflammatory effect (with a lower combination index CI value).

[0052] 5. Tests affecting melanin production

[0053] Human normal keratinocytes and melanocytes were seeded in 96-well plates at a ratio of 10:1, with a total seeding density of 1×10⁶ cells per well. 5 Cells / mL, cell volume 100 μL. After culturing for 24 h in a 37℃, 5% CO2 incubator, 100 µL of DMEM medium was added as a blank group, and samples containing single components or combinations of different proportions dissolved in DMEM medium were added as sample groups and cultured for 24 h. Then, they were exposed to 30 mJ / cm². 2 After UVB irradiation, the cells were cultured in fresh DMEM medium for 12 h.

[0054] After cell culture was completed, the cells were collected into centrifuge tubes and centrifuged at 1500 r / min for 10 min. The supernatant was discarded, and the cell pellet was washed with PBS. After centrifugation at 1500 r / min for 10 min, the supernatant was discarded, and 1.0 mol / L NaOH solution containing 10% DMSO was added. After incubation at 80℃ for 1 h, the melanin solution in the centrifuge tubes was transferred to a 96-well culture plate. The absorbance of each well was measured at a wavelength of 405 nm using a microplate reader to obtain the melanin inhibition rate.

[0055]

[0056] In equation (2): A1 and A2 represent the absorbance of the sample group and the blank group at 405 nm, respectively.

[0057] Based on the inhibitory effect test data of single components and compositions in different proportions, the combination index was calculated using Compusyn software. CI>1 indicates antagonistic effect, CI=1 indicates additive effect, and CI<1 indicates synergistic effect.

[0058] Table 4. Effects of dark tea extract on melanin production

[0059] Content of dark tea extract (μg / mg) Melanin inhibition rate (%) 0.5 3.4 1.0 14.3 1.5 24.8 2.0 36.9 2.5 47.9 3.0 55.7 3.5 64.9 4.0 70.8 4.5 79.5

[0060] Table 5. Effects of tranexamic acid on melanin production

[0061] Tranexamic acid content (μg / mg) Melanin inhibition rate (%) 0.5 1.9 1.0 5.7 2.0 9.2 4.0 16.6 8.0 23.5 16.0 36.5 24.0 48.7 30.0 53.9

[0062] Table 6. Effects of the composition on melanin production

[0063] Content of dark tea extract (μg / mg) Tranexamic acid content (μg / mg) Melanin inhibition rate (%) CI value 0.5 0.5 8.2 0.898 0.5 1.0 12.8 0.805 0.5 2.0 20.7 0.707 0.5 4.0 35.3 0.563 0.5 8.0 45.9 0.586 0.5 16.0 57.8 0.615 0.5 24.0 62.5 0.697 0.5 30.0 65.8 0.725

[0064] The results showed that both black tea extract and tranexamic acid alone could inhibit UVB-induced melanin production in keratinocytes. The combined index of different ratios (1:1~1:60) of the compositions calculated using Compusyn software was all <1, indicating that the compositions have synergistic effects, and that the combination of the two is necessary. Moreover, the ratio of black tea extract to tranexamic acid in the compositions of 1:8~1:32 had a better inhibitory effect on melanin production, which was roughly consistent with the results of the anti-inflammatory effects of different ratios of the compositions.

[0065] 6. Human efficacy test

[0066] Thirty-three volunteers (aged 18–50 years, with Individual Type Angle (ITA°) values ​​at the test sites ranging from 20° to 41°) were selected after informed consent. Exclusion criteria included: ① pregnant or breastfeeding women; ② individuals with skin allergies to cosmetics; ③ individuals with dermatitis or other skin conditions at the test site; ④ individuals who participated in other cosmetic efficacy tests within the past two months; ⑤ individuals who received any form of skin treatment within the past six months; ⑥ individuals who used corticosteroids or immunosuppressants at the test site or systemically during the trial; and ⑦ individuals who could not strictly adhere to the trial requirements or cooperate with the trial.

[0067] The VISIA-CR facial imaging system (CK GmbH, Germany) was used to collect data on the percentage of red area on the cheekbone region of volunteers before (D0) and after 14 (D14) and 28 (D28) days of use.

[0068] The whitening efficacy test was conducted under the conditions of (21±2)℃ and (50±10)% RH relative humidity. The whitening efficacy test method using the ultraviolet-induced human skin melanization model was adopted, namely: the first method of the whitening efficacy test method for cosmetics in the human efficacy evaluation test method newly added after the revision of the "Cosmetic Safety Technical Specifications" (2015 edition) on May 1, 2021 [Announcement of the National Medical Products Administration on Incorporating 7 Test Methods including the Test Method for Preservatives in Cosmetics into the Cosmetic Safety Technical Specifications (2015 Edition) (No. 17 of 2021)].

[0069] The test results are shown in the table. Table 9, of which Table 7 The composition in Table 9 is composed of black tea extract and tranexamic acid in a weight ratio of 1:16.

[0070] Table 7. Results of facial red area percentage measurement (mean ± standard deviation)

[0071]

[0072] Table 8. Results of melanin MI value measurement (mean ± standard deviation)

[0073]

[0074] Table 9. Skin Color ITA 。 Value measurement results (mean ± standard deviation)

[0075]

[0076] According to Table 7-9 of the human trial results, after 4 weeks of using a basic formula whitening and spot-removing essence containing single ingredients and combinations (the formula composition is: wt% 3% 1,3-butanediol, 3% glycerin, 1% black tea extract or tranexamic acid or a combination of both, 0.5% ethylhexylglycerin, 0.2% carbomer-941, 0.3% triethanolamine and the balance deionized water), the proportion of facial redness area was significantly reduced compared to using an essence containing a single ingredient, indicating that the combination has a better soothing effect; compared with the darkened skin, the individual type angle (ITA°) value increased and the melanin index (MI) value decreased, indicating that using a whitening and spot-removing essence containing combinations has a better whitening and spot-removing effect than using a single component.

[0077] 7. Tests on the combination of Cornus officinalis fruit extract and other compounds for melanin removal.

[0078] Cornus officinalis fruit extract: Fresh Cornus officinalis fruit was mixed with 5.5 times its weight of water to obtain a pulp, which was then sterilized. 1 wt% sucrose and Bifidobacterium animalis CICC 21711 (added at a rate of 2.5 × 10⁻⁶) were added to the pulp. 10 The mixture was fermented anaerobicly at 35°C for 65 hours (1 cell / g slurry), then sterilized to obtain the fermented product. The product was first concentrated under reduced pressure to 1.12 times the weight of the Cornus officinalis fruit. Then, a 35wt% ethanol solution of 15 times the weight of the concentrate was added, and the mixture was heated to 50°C and stirred at 100 rpm for 6 hours to obtain the extract. The extract was filtered through a 600-mesh filter, and the resulting coarse filtrate was filtered successively through 10, 0.8, 0.45, and 0.22 μm filter membranes to obtain the filtrate, which is the Cornus officinalis fruit extract.

[0079] Composition 1: Composed of black tea extract and tranexamic acid in a weight ratio of 1:8.

[0080] The effects of different test samples 1-9 (Cornus officinalis fruit extract, composition 1, and combinations of Cornus officinalis extract and composition 1 in different proportions) on inhibiting melanin production were compared. The composition of each test sample group is shown in Table 10. The A value in Table 10 represents the weight ratio of Cornus officinalis extract to composition 1, and the total concentration in the culture medium represents the total concentration of the active ingredients (Cornus officinalis extract and / or composition 1) in the culture medium.

[0081] Human normal keratinocytes and melanocytes were seeded in 96-well plates at a ratio of 10:1, with a total seeding density of 1×10⁶ cells per well. 5 Cells / mL, cell volume 100 μL. After culturing in a 37℃, 5% CO2 incubator for 24 h, samples containing DMEM medium (samples 1-9) were added and cultured for another 24 h. Then, they were exposed to 30 mJ / cm³. 2After UVB irradiation, the cells were cultured in fresh DMEM medium for 12 h.

[0082] After cell culture was completed, the cells were collected into centrifuge tubes and centrifuged at 1500 r / min for 10 min. The supernatant was discarded, and the cell pellet was washed with PBS. After centrifugation at 1500 r / min for 10 min, the supernatant was discarded, and 1.0 mol / L NaOH solution containing 10% DMSO was added. After incubation at 80℃ for 1 h, the melanin solution in the centrifuge tubes was transferred to a 96-well culture plate. The absorbance of each well was measured at a wavelength of 405 nm using a microplate reader to obtain the melanin inhibition rate. The results are shown in Table 10.

[0083] Table 10. Effects on melanin production

[0084] Sample number A value Total concentration in culture medium / μg / mg Melanin inhibition rate (%) Sample 1 1:0 0.45 32.5 Sample 2 0:1 0.45 35.3 Sample 3 1:0.1 0.45 33.1 Sample 4 1:0.3 0.45 35.8 Sample 5 1:0.6 0.45 39.7 Sample 6 1:0.7 0.45 45.2 Sample 7 1:0.85 0.45 42.4 Sample 8 1:1.7 0.45 36.6 Sample 9 1:4 0.45 34.0

[0085] According to the test results in Table 10, while maintaining a consistent total concentration, samples 4-8 (combination of composition 1 and Cornus officinalis extract) showed better melanin inhibition than samples 1-2 (cornus officinalis extract and composition 1). This indicates that the combination of black tea extract and tranexamic acid, when combined with Cornus officinalis extract, can synergistically enhance the melanin inhibition effect. Samples 5-7, which combine composition and Cornus officinalis extract, showed better results than samples 3-4 and 8-9. Therefore, when combining black tea extract and tranexamic acid with Cornus officinalis, a weight ratio of 1:0.6-0.85 yields the optimal melanin inhibition effect.

[0086] The specific embodiments described above are merely preferred embodiments to fully illustrate the present invention, and the scope of protection of the present invention is not limited thereto. Equivalent substitutions or modifications made by those skilled in the art based on the present invention are all within the scope of protection of the present invention. The scope of protection of the present invention is defined by the claims.

Claims

1. A composition having a whitening effect, characterized by comprising: The composition consists of black tea extract and tranexamic acid; The mass ratio of dark tea extract to tranexamic acid is 1:0.8-20; The preparation method of dark tea extract is as follows: mix dark tea with a polyol solution, heat and extract, and then filter. The polyol solution is composed of water and polyol in a volume ratio of (5-7):(2-3.5); The polyol is 1,3-butanediol.

2. The composition with whitening effect according to claim 1, characterized in that, The polyol solution is composed of water and polyol in a volume ratio of 7:

3.

3. The composition with whitening effect according to claim 1, characterized in that, The temperature and time for heating and extraction are 50-60℃ and 30-200 min.

4. A method for preparing a composition with whitening effect according to any one of claims 1-3, characterized in that, This includes the steps of mixing and packaging the raw materials.

5. The application of a composition with whitening effect according to any one of claims 1-3 in the preparation of cosmetics, characterized in that, The composition having whitening effect is present in the cosmetic at a weight percentage of 1.5-8.0%.

6. The application according to claim 5, characterized in that, The cosmetic also includes 2-10% Cornus officinalis fruit extract, with the mass ratio of the Cornus officinalis fruit extract to the composition having whitening effect being 1:0.6-0.

85. The Cornus officinalis fruit extract is obtained by pulping Cornus officinalis fruit with water, sterilizing, adding sucrose and Bifidobacterium for fermentation, sterilizing again, concentrating for the first time, and then using 20-45wt% ethanol solution as the extraction solvent for heating and extraction, followed by filtration to obtain the filtrate, which is the Cornus officinalis fruit extract.

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