A composition for lightening dark circles under the eyes containing an extract of camellia japonica and a method for preparing and using the same

Abstract

The application discloses a composition for lightening dark circles under eyes containing camellia chrysantha extract and a preparation method and application thereof, and relates to the technical field of cosmetics.The composition comprises component A, component B and component C;the component A comprises andrographis paniculata extract and camellia chrysantha extract;the component B comprises ubiquinone and safflower extract;and the component C is menthol lactate.The andrographis paniculata extract, the camellia chrysantha extract, the ubiquinone, the safflower extract and the menthol lactate are combined to have a synergistic effect; the composition formed by the specific combination of the components can promote blood microcirculation, inhibit microvascular generation, increase epidermal thickness, relieve cell hypoxia and eye fatigue, reduce blood vessel type dark circles, relieve redness and strengthen skin, and is mild and non-irritating to skin, and is suitable for people with sensitive skin.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, and in particular to a composition for lightening dark circles containing Camellia chrysantha extract, its preparation method, and its application. Background Technology

[0002] Many young people today suffer from dark circles under their eyes due to factors such as staying up late, high stress, and excessive screen time. The majority of these are vascular dark circles. Vascular dark circles are mainly caused by factors such as rhinitis, lack of sleep, eye fatigue, endocrine disorders, and anemia, leading to stagnation of blood circulation around the eyes, or by thinning and increasing the transparency of the lower eyelid skin, making the blood vessels clearly visible. These dark circles appear blue, pink, or purple, and may be accompanied by swelling around the eyes.

[0003] Among existing technologies, cosmetic procedures such as phototherapy can effectively shrink and close blood vessels, improving vascular dark circles; injectable treatments can increase the thickness of the lower eyelids and reduce blood vessel exposure. While these methods offer quick results, they may carry risks, have side effects, and are expensive. Common active ingredients in eye creams, such as caffeine and vitamin K, promote blood circulation, but both have some skin irritation, offering only mild effects with short-lived results, making it difficult to fundamentally improve vascular dark circles. Therefore, many existing technologies and products suffer from drawbacks such as irritating ingredients, slow effects, limitations, and high costs, and are often unsuitable for different skin types.

[0004] Therefore, products that soothe and repair vascular dark circles have become a pressing technical problem to be solved in this field. Summary of the Invention

[0005] Based on this, the purpose of the present invention is to overcome the shortcomings of the prior art and provide a composition for lightening dark circles containing Camellia chrysantha extract and its application. This composition can reduce vascular dark circles, soothe redness, and strengthen the skin by promoting blood microcirculation, inhibiting microvascular formation, increasing epidermal thickness, relieving cell hypoxia and eye fatigue. Moreover, it is gentle and non-irritating to the skin and suitable for people with sensitive skin.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0007] In a first aspect, the present invention provides a composition for lightening dark circles, the composition comprising component A, component B and component C; component A comprising andrographis paniculata extract and Camellia chrysantha extract; component B comprising ubiquinone and saffron extract; and component C being menthol lactate.

[0008] Andrographis paniculata extract is rich in terpenoids such as andrographolide, which inhibit angiogenesis and dilation, thereby reducing microvessels under the eyes. Camellia chrysantha extract is rich in flavonoids and polyphenols, which promote skin cell repair and regeneration, increasing epidermal thickness; long-term use can reduce the visibility of blood vessels around the eyes. Ubiquinone, also known as coenzyme Q10, is a coenzyme for cellular aerobic respiration, enhancing cellular energy metabolism, promoting oxygen utilization, and alleviating hypoxia caused by poor blood circulation under the eyes. Saffron extract has the effects of promoting blood circulation, removing blood stasis, and relieving stagnation, promoting microcirculation, and alleviating blood stasis under the eyes caused by staying up late, stress, and emotions. Menthol lactate is a menthol derivative with a mild minty aroma, a long-lasting cooling sensation, and no burning or irritation; it provides a gentle and lasting cooling sensation to the skin, causing vasoconstriction and effectively reducing the burning and soreness in the eyes caused by fatigue.

[0009] This invention has found that when the above-mentioned plant extracts are combined with ubiquinone and menthol lactate, they can promote blood microcirculation, inhibit microvascular formation, increase epidermal thickness, relieve cellular hypoxia and eye fatigue, thereby reducing vascular dark circles, soothing redness, and strengthening the skin. Moreover, they are gentle and non-irritating to the skin, making them suitable for people with sensitive skin.

[0010] As a preferred embodiment of the first aspect, the mass ratio of component A, component B and component C in the composition is component A: component B: component C = (0.6-10):(0.11-5.5):(0.001-0.05).

[0011] As a preferred embodiment of the first aspect, the mass ratio of component A, component B and component C in the composition is component A: component B: component C = (1.5-7):(0.55-3.4):(0.005-0.03).

[0012] In a preferred embodiment of the first aspect, the mass ratio of component A, component B and component C in the composition is component A: component B: component C = 4:2.2:0.01.

[0013] This invention has found that the mass ratio of components A, B, and C in a dark circle-lightening composition not only affects the VEGF reduction rate, filaggrin increase rate, and epidermal thickness increase rate, thus influencing the composition's ability to lighten blood vessels and strengthen the skin, but also its improvement on redness a* value, moisture content, and dark circle area reduction rate, thereby affecting the composition's ability to lighten dark circles, soothe redness, and moisturize. When components A, B, and C are further selected at (0.6-10):(0.11-5.5):(0.001-0.05), especially (1.5-7):(0.55-3.4):(0.005-0.03), the resulting dark circle-lightening composition exhibits superior functional effects in lightening blood vessels, strengthening the skin, lightening dark circles, soothing, and moisturizing; thus significantly enhancing the overall comprehensive effect of the composition. Research has shown that the overall effect of the composition of the present invention is optimal when the mass ratio of component A, component B and component C is 4:2.2:0.01.

[0014] In a preferred embodiment of the first aspect, the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract in component A is Andrographis paniculata extract: Camellia chrysantha extract = (0.1-5):(0.5-5).

[0015] In a preferred embodiment of the first aspect, the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract in component A is Andrographis paniculata extract: Camellia chrysantha extract = (0.5-3):(1-4).

[0016] In a preferred embodiment of the first aspect, the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract in component A is Andrographis paniculata extract: Camellia chrysantha extract = 2:2.

[0017] This invention has found that the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract in component A of the dark circle-lightening composition can, to some extent, affect the production of vascular endothelial growth factor (VEGF) and the growth of filaggrin and epidermis in the skin, thereby influencing the composition's ability to lighten blood vessels and strengthen the skin. Further selection of the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract at (0.1-5):(0.5-5), especially (0.5-3):(1-4), can enhance the composition's efficacy in lightening blood vessels and strengthening the skin, thus improving the overall comprehensive ability of the composition to improve vascular dark circles. Research has shown that when the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract is 2:2, the composition of this invention exhibits the best ability to lighten blood vessels and strengthen the skin.

[0018] In a preferred embodiment of the first aspect, the mass ratio of ubiquinone to saffron extract in component B is ubiquinone:saffron extract = (0.01-0.5):(0.1-5).

[0019] In a preferred embodiment of the first aspect, the mass ratio of ubiquinone to saffron extract in component B is ubiquinone:saffron extract = (0.05-0.4):(0.5-3).

[0020] In a preferred embodiment of the first aspect, the mass ratio of ubiquinone to saffron extract in component B is ubiquinone:saffron extract = 0.3:1.9.

[0021] This invention has found that the mass ratio of ubiquinone to saffron extract in component B of the composition for fading dark circles can, to some extent, affect blood circulation and blood stasis in the under-eye skin, thus impacting the composition's ability to improve dark circles. Further selection of the mass ratio of ubiquinone to saffron extract to (0.01-0.5):(0.1-5), especially (0.05-0.4):(0.5-3), enhances the composition's effects of promoting blood circulation, removing blood stasis, and relieving stagnation, thereby improving the microcirculation of the under-eye skin, fading dark circles, and strengthening the overall comprehensive ability of the composition to improve vascular dark circles. Research has shown that when the mass ratio of ubiquinone to saffron extract is 0.3:1.9, the composition of this invention exhibits the best ability to promote blood circulation, remove blood stasis, relieve stagnation, and fade dark circles.

[0022] As a preferred embodiment of the first aspect, the method for preparing the Camellia chrysantha extract includes the following steps: drying Camellia chrysantha flowers, pulverizing them, adding them to an aqueous solution of a eutectic solvent, ultrasonically extracting them, collecting the product, centrifuging, and taking the supernatant to obtain the Camellia chrysantha extract; wherein, the eutectic solvent is composed of choline chloride solution and 1,3-propanediol.

[0023] As a preferred embodiment of the first aspect, the method for preparing the Camellia chrysantha extract satisfies at least one of the following:

[0024] (a) The mass ratio of choline chloride to 1,3-propanediol is choline chloride: 1,3-propanediol = 1:(1-5);

[0025] (b) The aqueous solution of the eutectic solvent has a mass concentration of 60–90%;

[0026] (c) The temperature of the ultrasonic extraction is 40-60℃; or / and the power of the ultrasonic extraction is 120-200W, and the time of the ultrasonic extraction is 20-50min;

[0027] (d) The material-to-liquid ratio for ultrasonic extraction is 1g:(20-40)mL.

[0028] This invention has found that different extraction methods affect the extract of Camellia chrysantha and its synergistic effect with other components, thereby affecting the composition's ability to lighten blood vessels, improve skin blood circulation and strengthen the skin, and thus affecting the overall efficacy of the composition in lightening dark circles.

[0029] In a second aspect, the present invention provides the use of the dark circle-reducing composition as described in the first aspect in the preparation of cosmetics.

[0030] Thirdly, the present invention provides an eye cream comprising the following components by weight percentage: 1-15% of the composition, 0.05-0.5% of thickener, 0.5-2% of moisturizer, 5-20% of emulsifier, 0.5-3% of preservative, 0.01-0.3% of pH adjuster, and the balance being deionized water.

[0031] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0032] This invention discovers that extracts of Andrographis paniculata, Camellia chrysantha, and Saffron, when used in combination with ubiquinone and menthol lactate, have a synergistic effect. The composition formed by these extracts through specific formulation can promote blood microcirculation, inhibit microvascular formation, increase epidermal thickness, relieve cellular hypoxia and eye fatigue, thereby reducing vascular dark circles, soothing redness, and strengthening the skin. Moreover, it is gentle and non-irritating to the skin, making it suitable for people with sensitive skin. Attached Figure Description

[0033] Figure 1 This is a comparison chart showing the improvement in the area of ​​vascular dark circles under the eyes of volunteers before and after use in the blank application example and application example 15 of this invention. Detailed Implementation

[0034] To better illustrate the purpose, technical solution, and advantages of this invention, the invention will be further described below with reference to specific embodiments. The purpose is to provide a detailed understanding of the invention, not to limit it. All other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention. Unless otherwise specified, the experimental reagents and instruments designed in the embodiments and comparative examples of this invention are commonly used reagents and instruments, all of which are commercially available. Unless otherwise specified, the experimental methods used in the embodiments and comparative examples are conventional methods; and unless otherwise specified, the raw materials used in parallel experiments are from the same batch. Further explanation of the raw materials used in this invention:

[0035] Andrographis paniculata extract was purchased from Shaanxi Huike Plant Development Co., Ltd., product number: HKY002;

[0036] Saffron extract was purchased from Guangzhou Fanhua Biotechnology Co., Ltd., product number: 20220321;

[0037] Ubiquinone was purchased from Shanxi Qixin Biotechnology Co., Ltd., product number: QX-0516-034;

[0038] Menthol lactate was purchased from Luyuan Natural Fragrance Oil Refinery in Qingyuan District, Ji'an City, item number: WS-23;

[0039] White water lily extract was purchased from Shaanxi Shize Tiancheng Biotechnology Co., Ltd., product number: BSLTQW01;

[0040] Camellia extract was purchased from Xi'an Wanfang Biotechnology Co., Ltd., product number: WFSW-03;

[0041] Safflower extract was purchased from Shaanxi Shize Tiancheng Biotechnology Co., Ltd., product number: HHTQY01;

[0042] Menaquinone was purchased from Shandong Yijia Trading Co., Ltd., item number: 198;

[0043] Menthol was purchased from Luyuan Natural Fragrance Oil Refinery in Qingyuan District, Ji'an City, product number: Menthol Ice;

[0044] The preparation method of Camellia chrysantha extract-1 is as follows:

[0045] (1) Prepare a eutectic solvent:

[0046] Dissolve 30g of choline chloride in 90g of 1,3-propanediol to obtain a eutectic solvent; dissolve 70g of the eutectic solvent in 30g of deionized water to obtain an aqueous solution of the eutectic solvent with a mass concentration of 70%.

[0047] (2) Preparation of Camellia chrysantha extract:

[0048] Flowers of Camellia chrysantha were collected, cleaned, and dried in a 50°C constant temperature oven until constant weight. After grinding and pulverizing, the powder was passed through an 80-mesh sieve. 1g of the powder was added to an aqueous solution of the above-mentioned eutectic solvent at a ratio of 1g:30mL. Ultrasonic extraction was performed at a temperature of 40°C, a power of 150W, and an extraction time of 30min. The product was collected, centrifuged, and the supernatant was taken to obtain Camellia chrysantha extract-1.

[0049] The preparation method of Camellia chrysantha extract-2 is as follows:

[0050] (1) Prepare a eutectic solvent:

[0051] Dissolve 30g of choline chloride in 30g of 1,3-propanediol to obtain a eutectic solvent. Dissolve 60g of the eutectic solvent in 40g of deionized water to obtain an aqueous solution of the eutectic solvent with a mass concentration of 60%.

[0052] (2) Preparation of Camellia chrysantha extract:

[0053] Flowers of *Camellia chrysantha* were collected, cleaned, and dried in a 50°C oven until constant weight. The powder was then ground and passed through an 80-mesh sieve. 1 g of the powder was added to an aqueous solution of the aforementioned eutectic solvent at a ratio of 1 g to 40 mL. Ultrasonic extraction was performed at 50°C, 120 W, and for 50 min. The product was collected, centrifuged, and the supernatant was taken to obtain *Camellia chrysantha* extract-2.

[0054] The preparation method of Camellia chrysantha extract-3 is as follows:

[0055] (1) Prepare a eutectic solvent:

[0056] Dissolve 30g of choline chloride in 150g of 1,3-propanediol to obtain a eutectic solvent. Dissolve 90g of the eutectic solvent in 10g of deionized water to obtain an aqueous solution of the eutectic solvent with a mass concentration of 90%.

[0057] (2) Preparation of Camellia chrysantha extract:

[0058] Flowers of *Camellia chrysantha* were collected, cleaned, and dried in a 50°C oven until constant weight. The powder was then ground and passed through an 80-mesh sieve. 1 g of the powder was added to an aqueous solution of the aforementioned eutectic solvent at a ratio of 1 g to 20 mL. Ultrasonic extraction was performed at 60°C, 200 W, and for 20 min. The product was collected, centrifuged, and the supernatant was taken to obtain *Camellia chrysantha* extract-3.

[0059] The preparation method of Camellia chrysantha extract-4 is as follows:

[0060] Flowers of Camellia chrysantha were collected, cleaned, and dried in a 50°C constant temperature oven until constant weight. After grinding and pulverizing, the powder was passed through an 80-mesh sieve. 1g of the powder was added to a 75% ethanol aqueous solution at a material-to-liquid ratio of 1g:30mL. Ultrasonic extraction was performed at 50°C, 180W, and 30min. The product was collected, centrifuged, and the supernatant was collected to obtain Camellia chrysantha extract-4.

[0061] The preparation method of Camellia chrysantha extract-5 is as follows:

[0062] Flowers of Camellia chrysantha were collected, cleaned, and dried in a 50°C constant temperature oven until constant weight. After grinding and pulverizing, the powder was passed through an 80-mesh sieve. 1g of the powder was taken and pure water was added, with a material-to-liquid ratio of 1g:40mL. Ultrasonic extraction was performed at a temperature of 50°C, a power of 180W, and an extraction time of 30min. The product was collected, centrifuged, and the supernatant was taken to obtain Camellia chrysantha extract-5.

[0063] The preparation method of the composition for lightening dark circles of the present invention is as follows: weigh each component by weight, stir and mix evenly, and sonicate for 10 minutes to obtain the composition for lightening dark circles.

[0064] Examples and Comparative Examples

[0065] The examples and comparative examples provide a composition for lightening dark circles, the components and their weight parts of which are shown in Table 1:

[0066] Table 1:

[0067]

[0068]

[0069] Application examples

[0070] This application example provides an eye cream, the components of which (mass percentage) are shown in Table 2. The compositions used in Application Examples 1-19 are the dark circle-lightening compositions prepared in Examples 1-19. The compositions used in Comparative Application Examples 1-12 are the compositions prepared in Comparative Examples 1-12. For example, the dark circle-lightening composition used in Application Example 1 is the dark circle-lightening composition from Example 1, and so on. The compositions used in Application Examples 20-21 are the dark circle-lightening compositions prepared in Example 15. The compositions used in Comparative Application Examples 1-12 are the dark circle-lightening compositions prepared in Comparative Examples 1-12. The difference between the blank application examples and the application examples is that the compositions of this invention are not added. See Table 2 for details.

[0071] Table 2:

[0072]

[0073] This application example also provides a method for preparing the above-mentioned eye cream, including the following steps:

[0074] (1) Mix the humectant and thickener with water and stir. Heat to 85±2℃ and homogenize at 1200rpm for 4min. After homogenization, keep warm for later use to obtain the pre-prepared phase A.

[0075] (2) Mix the emulsifier, heat to 85±2℃, and homogenize at 1200rpm for 4min. After homogenization, keep warm for later use to obtain the pre-prepared phase B.

[0076] (3) Mix the preservatives and heat to 60±2℃ to melt them to obtain the pre-prepared C phase;

[0077] (4) Heat the pre-prepared phase A to 80±2℃, add the pre-prepared phase B at 250 rpm, stir and mix, then cool down to 60±2℃, add the pre-prepared phase C at 250 rpm and stir and mix, then cool down to 40℃, add the dark circle lightening composition and continue stirring for 8 minutes, finally add the remaining pH adjuster to adjust the pH to 6.0, then stop stirring, discharge the material, and obtain the eye cream.

[0078] Example 1: Cell experiments demonstrated that the composition for lightening dark circles can inhibit vasodilation.

[0079] This study investigates the angiogenesis inhibition effect of the dark circle-lightening compositions prepared in Examples 1-19 and Comparative Examples 1-12:

[0080] Experimental Methods: The cell line used was human skin capillary endothelial cells (Beina Biotechnology). The test conditions were: incubator temperature 37±1℃, saturated humidity, and carbon dioxide 5±1%. Cells were cultured and treated according to groups, followed by testing. The specific test was the determination of vascular endothelial growth factor (VEGF) content, and the test method is as follows:

[0081] (1) The cell suspension was seeded into a 96-well cell culture plate at a density of 2000 cells / well. 100 μL of HDMEC cell culture medium (Beina Biotechnology) was added to each well and cultured for 24 h.

[0082] (2) Discard the supernatant, add 100 μL of culture medium to the control group, and add 100 μL of culture medium containing 0.1% of the corresponding sample (Examples 1-19 and Comparative Examples 1-12) to the experimental group, and incubate for 24 h;

[0083] (3) Collect cells separately, wash 3 times with PBS buffer, add 50 μL of RIPA cell lysis buffer containing protease inhibitor to each well, centrifuge the lysed sample at 12000g for 5 minutes, and take the supernatant.

[0084] (4) Determination of vascular endothelial growth factor (VEGF) content: The VEGF content in the cell supernatant of each group was detected using a human vascular endothelial growth factor A (VEGF-A) ELISA kit (catalog number BLL108277E, Baililai Biotechnology); the reduction rate of VEGF in each example and comparative example was calculated using the following formula:

[0085] VEGF reduction rate (%) = (content of blank group - content of sample group) / content of blank group * 100%. The higher the VEGF reduction rate, the stronger the ability of the composition to inhibit blood vessels. The results are shown in Table 3.

[0086] Table 3

[0087]

[0088]

[0089] As can be seen from Table 3, the composition for lightening dark circles provided by the present invention reduces VEGF secretion by more than 47%, which indicates that the composition of the present invention has a strong effect in inhibiting angiogenesis.

[0090] As can be seen from Examples 1-15 and Comparative Examples 1-2, the mass ratio of the components in the composition for lightening dark circles affects the composition's ability to inhibit VEGF, thereby affecting the composition's effect on inhibiting angiogenesis.

[0091] Examples 1-5 show that the mass ratio of the components in the composition for lightening dark circles affects the composition's ability to inhibit VEGF secretion. When the mass fractions of the components are limited to the range of (0.6-10):(0.11-5.5):(0.001-0.05) for component A, component B, and component C, the resulting composition exhibits good anti-angiogenic effects. When the mass ratio of component A, component B, and component C is further selected to be within the range of (1.5-7):(0.55-3.4):(0.005-0.03), the anti-angiogenic effect of the resulting composition is even better. The best effect is achieved when the mass ratio of component A, component B, and component C is 4:2.2:0.01 (Example 5).

[0092] This invention investigated the effect of the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract in component A on the ability of the composition to inhibit VEGF secretion through Examples 6-8. Examples 6-8 show that the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract affects the composition's effect on inhibiting VEGF secretion, thereby affecting its anti-angiogenic efficacy. When the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract is limited to the range of (0.1-5):(0.5-5), the composition's ability to inhibit VEGF secretion is improved. Further optimization by limiting the mass ratio to the range of (0.5-3):(1-4) yields an even better composition. The optimal effect is achieved when the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract is 2:2 (Example 7).

[0093] This invention investigated the effects of different extraction methods of Camellia chrysantha extract obtained from component A on the ability of the composition to inhibit VEGF secretion through Examples 15-19. Examples 15-19 show that the extraction method of Camellia chrysantha extract has a certain influence on the effect of inhibiting VEGF secretion, thus affecting the composition's anti-angiogenic efficacy. The composition obtained using Camellia chrysantha extract-1 showed the best anti-angiogenic effect (Example 15).

[0094] The VEGF reduction rate of Comparative Examples 1-2 was 19-23%. This is because the mass ratio of each component in the compositions of Comparative Examples 1-2 was not within the range of the present invention, which affected the synergistic effect between the components and reduced the efficacy of the composition.

[0095] The VEGF reduction rate of Comparative Examples 3-7 was 8-21%, which is because the absence of one of the components of the present invention in the compositions of Comparative Examples 3-7 affected the synergistic effect between the components and reduced the efficacy of the composition.

[0096] The VEGF reduction rate of Comparative Examples 8-12 was 5-17%. This is because the compositions of Comparative Examples 8-12 replaced one of the components in the composition of the present invention with other plant extracts, which did not produce a synergistic effect with the specific components selected in the present invention, and also affected the synergistic effect between other components of the present invention, thus reducing the efficacy of the composition.

[0097] Example 2: Cell experiments demonstrated that the prepared eye cream could increase epidermal thickness.

[0098] This study investigates the repairing effects of eye creams prepared using Examples 1-21, compared with Examples 1-12, and the blank example.

[0099] Experimental Methods: The cell model used was a human 3D epidermal skin model (Guangdong Boxi Biotechnology Co., Ltd.). The test conditions were: incubator temperature 37±1℃, saturated humidity, and carbon dioxide 5±1%. The 3D epidermal models were cultured and treated according to groups, followed by testing. Specifically, the filaggrin (FLG) content and epidermal thickness were measured using the following methods:

[0100] (1) The 3D epidermal skin model was cultured using EpiGrowth medium (Guangdong Boxi Biotechnology Co., Ltd.);

[0101] (2) Apply 25 μL of 0.1% sodium dodecyl sulfate (Sigma) to the surface of the 3D epidermal skin model and incubate for 30 minutes;

[0102] (3) In the blank control group, 12.5 μL of the eye cream of the blank application example was evenly applied to the surface of the epidermal model. In the sample group, 12.5 μL of the eye cream of application example 1-21 and control application example 1-18 were applied and cultured for 24 hours.

[0103] (4) After the culture was completed, the test substance remaining on the model surface was cleaned with PBS solution (Biosharp), fixed with 4% paraformaldehyde (Biosharp) for 24 h and then circumcised.

[0104] (5) Detection of filaggrin FLG content: The tissue was lysed with RIPA cell lysis buffer containing protease inhibitor. The lysed sample was centrifuged at 12000g for 5 minutes. The supernatant was collected and the filaggrin FLG content in the supernatant of each group of tissues was detected using a human filaggrin (FLG) ELISA kit (catalog number BLL107115E, Baililai Biotechnology).

[0105] Filaggrin (FLG) is an important molecule in the stratum corneum of human skin that connects keratin fibers. With the assistance of FLG monomers, keratin fibers aggregate regularly, forming a solid physical barrier on the outermost layer of the epidermis, thus strengthening the skin.

[0106] (6) Epidermal thickness measurement: The tissue sections were stained with H&E (Beyotime), photographed and observed under a microscope, and the images were collected and analyzed.

[0107] The increase rate of filaggrin and epidermal thickness in each example and comparative example was calculated using the following formula:

[0108] Increase rate (%) = (Sample group - Blank group) / Blank group * 100%. A higher increase rate in filaggrin indicates a higher increase in epidermal thickness, signifying a stronger skin-strengthening effect of the eye cream. Results are shown in Table 4.

[0109] Table 4:

[0110]

[0111]

[0112] As can be seen from Table 4, the eye cream for reducing vascular dark circles provided by the present invention increases filaggrin by more than 43% and increases epidermal thickness by more than 19%. This indicates that the eye cream with the added composition of the present invention can increase epidermal thickness and strengthen the skin.

[0113] As can be seen from Application Examples 1-15 and Comparative Application Examples 1-2, the mass ratio of the components in the composition affects the composition's ability to generate skin filaggrin and increase epidermal thickness, thereby affecting the composition's skin-strengthening effect.

[0114] As can be seen from Application Examples 1-5, the mass ratio of the components in the composition affects the composition's ability to promote filaggrin formation and contributes to the increase in epidermal thickness. When the mass fractions of the components are limited to the range of (0.6-10):(0.11-5.5):(0.001-0.05) for component A, component B, and component C, the resulting composition has a good effect on promoting filaggrin formation. When the mass ratio of component A, component B, and component C is further selected to be in the range of (1.5-7):(0.55-3.4):(0.005-0.03), the effect on promoting filaggrin formation is even better. The best effect is achieved when the mass ratio of component A, component B, and component C is 4:2.2:0.01 (Application Example 5).

[0115] This invention investigated the effect of the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract in component A on the composition's ability to promote filaggrin formation through Application Examples 6-8. Application Examples 6-8 show that the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract affects the composition's effect on promoting filaggrin formation, thus influencing its skin-strengthening efficacy. When the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract is limited to the range of (0.1-5):(0.5-5), the composition's ability to promote filaggrin formation is improved. Further optimization by limiting the mass ratio to the range of (0.5-3):(1-4) yields an even better composition. The optimal effect is achieved when the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract is 2:2 (Application Example 7).

[0116] This invention investigated the effects of different extraction methods of *Camellia chrysantha* extracts obtained from component A on the ability of the composition to increase polyprotein content and epidermal thickness through Application Examples 15-19. Application Examples 15-19 show that the extraction method of *Camellia chrysantha* extract has a certain influence on the effect of increasing polyprotein content and epidermal thickness, thus affecting the composition's efficacy in increasing polyprotein content and epidermal thickness. The composition obtained using *Camellia chrysantha* extract-1 showed the best effect in increasing polyprotein content and epidermal thickness (Application Example 15).

[0117] Compared with Application Examples 1-2, the increase rate of filaggrin was 12%, and the increase rate of epidermal thickness was 3-7%. This is because the mass ratio of each component in the composition of Comparative Examples 1-2 was not within the scope of the present invention, which affected the synergistic effect between the components and reduced the efficacy of the composition.

[0118] Compared with the 6-18% increase in filaggrin and 2-8% increase in epidermal thickness in Comparative Examples 3-7, this is because the absence of one of the components of the present invention in the compositions of Comparative Examples 3-7 affected the synergistic effect between the components and reduced the efficacy of the composition.

[0119] Compared with the 6-20% increase in filaggrin and the 2-7% increase in epidermal thickness in Comparative Examples 8-12, this is because the compositions of Comparative Examples 8-12 replaced one of the components in the composition of the present invention with other plant extracts, which did not produce a synergistic effect with the specific components selected in the present invention, and also affected the synergistic effect between other components of the present invention, thus reducing the efficacy of the composition.

[0120] Example 3: Human skin patch test.

[0121] Thirty volunteers were recruited, 15 men and 15 women, aged 20-50 years. A closed patch test method was used. Equal volumes (0.020 mL-0.025 mL) of test samples (eye creams prepared for the application example, control example, and blank example) were placed in a specific patch applicator. The patch was then applied to the volunteer's arm with hypoallergenic adhesive tape, and gently pressed to ensure even application to the skin. The patch was left on for 24 hours. The blank control group used distilled water, and the blank application examples used eye creams without a dark circle-lightening composition. After 24 hours, the patch applicator was removed, and skin reactions were observed and recorded at 0.5 h, 24 h, and 48 h. The severity of adverse skin reactions is shown in Table 5 below.

[0122] Table 5:

[0123]

[0124] After testing, the eye creams for reducing dark circles provided in Application Examples 1-21 and Comparative Application Examples 1-12 of this invention showed negative reactions after human patch testing, indicating that they are safe and non-irritating to human skin.

[0125] Example 4: Human efficacy test for reducing vascular dark circles

[0126] This study investigates the effect of eye creams prepared in Application Examples 1-21, Comparative Application Examples 1-12, and Blank Application Examples on reducing vascular dark circles.

[0127] Experimental methods:

[0128] One hundred and seventy Asian adults aged 18-30 with sensitive skin and vascular dark circles were selected and randomly divided into 34 groups of five. Volunteers applied the sample to the eye area twice daily, morning and evening, and data were collected on day 0 and day 28.

[0129] After arriving, volunteers washed their faces with facial cleanser and sat quietly for 30 minutes in an air-conditioned room with a temperature of 21±1℃ and a humidity of 50±10%. Researchers used VISIA-CR to photograph and analyze dark circles under the eyes, used the CK skin color test probe CL400 to measure the redness a* value of the skin under the eyes, and used the skin moisture test probe Corneometer to measure the moisture content of the skin under the eyes.

[0130] The ability to reduce vascular dark circles is represented by the improvement in the area of ​​dark circles; the soothing ability is represented by the improvement in the redness (a*) value of the skin under the eyes; and the moisturizing ability is represented by the improvement in the moisture content of the skin under the eyes. The formulas are as follows:

[0131] Dark circle area improvement rate = (T0) 样品组 -T28 样品组 ) / T0 样品组 *100%-(T0) 空白应用例 -T28 空白应用例 ) / T0 空白应用例 *100%;

[0132] Redness a* Improvement Rate = (T0) 样品组 -T28 样品组 ) / T0 样品组 *100%-(T0) 空白应用例 -T28 空白应用例 ) / T0 空白应用例 *100%;

[0133] Moisture content improvement rate = (T28) 样品组 -T0 样品组 ) / T0 样品组 *100%-(T28) 空白应用例 -T0 空白应用例 ) / T0 空白应用例 *100%;

[0134] The higher the improvement rate of dark circle area, the stronger the eye cream's ability to improve or lighten dark circles; the higher the improvement rate of redness (a*), the stronger the eye cream's soothing ability; and the higher the improvement rate of moisture content, the stronger the eye cream's moisturizing ability. The results are shown in Table 6.

[0135] Table 6:

[0136] Group Dark circle area improvement rate % a* value improvement rate % Moisture content improvement rate % Application Example 1 29.2 15.5 23.3 Application Example 2 29.9 15.1 22.4 Application Example 3 34.1 17.2 26.2 Application Example 4 33.9 17.4 26.1 Application Example 5 36.7 18.5 28.2 Application Example 6 38.1 19.1 29.1 Application Example 7 40.0 20.1 30.5 Application Example 8 36.6 18.5 28.0 Application Example 9 35.5 17.8 27.2 Application Example 10 40.0 20.1 30.4 Application Example 11 34.9 17.6 26.8 Application Example 12 35.3 17.8 27.3 Application Example 13 34.1 17.2 26.3 Application Example 14 33.9 17.2 26.3 Application Example 15 46.4 23.9 35.5 Application Example 16 45.1 23.3 34.6 Application Example 17 45.1 23.4 34.7 Application Example 18 43.2 22.2 33.0 Application Example 19 42.2 21.6 32.3 Application Example 20 30.3 19.2 26.1 Application Example 21 43.4 20.3 34.5 Comparative Application Example 1 14.0 5.9 8.5 Comparative Application Example 2 9.6 4.9 4.3 Comparative Application Example 3 10.9 5.3 7.8 Comparative Application Example 4 12.7 6.5 11.2 Comparative Application Example 5 12.7 6.0 4.5 Comparative Application Example 6 10.3 3.3 3.8 Comparative Application Example 7 14.3 5.9 9.9 Comparative Application Example 8 9.8 4.8 5.9 Comparative Application Example 9 10.1 2.1 11.4 Comparative Application Example 10 8.0 2.5 9.2 Comparative Application Example 11 8.8 4.3 11.0 Comparative Application Example 12 5.3 7.3 2.5

[0137] As can be seen from Table 6, the eye cream for reducing vascular dark circles provided by the present invention reduced the dark circle area by more than 29%, the redness a* value by more than 15%, and the skin moisture content by more than 22% after 28 days of use. This indicates that the eye cream with the added composition of the present invention can play a comprehensive role in reducing dark circles, promoting skin blood circulation, soothing and moisturizing.

[0138] As can be seen from Application Examples 1-15 and Comparative Application Examples 1-2, the mass ratio of the components in the dark circle lightening composition affects the composition's ability to improve the area, moisture content, and a* value of dark circles, thereby affecting the composition's effects of lightening dark circles, soothing, and moisturizing.

[0139] As can be seen from Application Examples 1-5, the mass ratio of the components in the composition for reducing dark circles affects the composition's ability to reduce dark circles, promote skin blood circulation, soothe, and moisturize. When the mass ratio of components A, B, and C is limited to the range of (0.6-10):(0.11-5.5):(0.001-0.05), the resulting composition exhibits good effects in reducing dark circles, promoting skin blood circulation, soothing, and moisturizing. When the mass ratio of components A, B, and C is further selected to be within the range of (1.5-7):(0.55-3.4):(0.005-0.03), the effect of the resulting composition is even better. The best effect is achieved when the mass ratio of components A, B, and C is 4:2.2:0.01 (Application Example 5).

[0140] This invention investigated the effect of the mass ratio of Andrographis paniculata extract and Camellia chrysantha extract in component A on the composition's efficacy in reducing dark circles, soothing, and moisturizing through Application Examples 6-8. Application Examples 6-8 show that the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract affects the composition's ability to reduce dark circles, soothe, and moisturize. When the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract is limited to the range of (0.1-5):(0.5-5), the improvement rate of dark circle area, the improvement rate of a* value, and the moisture content are increased, thus enhancing the composition's ability to reduce dark circles, soothe, and moisturize. Further optimization by limiting the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract to the range of (0.5-3):(1-4) yields even better results. The optimal effect is achieved when the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract is 2:2 (Application Example 7).

[0141] This invention investigated the effect of the mass ratio of ubiquinone to saffron extract in component B on the composition's efficacy in reducing dark circles, soothing, and moisturizing through Application Examples 9-11. Application Examples 9-11 show that the mass ratio of ubiquinone to saffron extract affects the composition's ability to reduce dark circles, soothe, and moisturize. When the mass ratio of ubiquinone to saffron extract is limited to the range of (0.01-0.5):(0.1-5), the improvement rate of dark circle area, the improvement rate of a* value, and the moisture content are increased, thus improving the composition's ability to reduce dark circles, soothe, and moisturize. Further optimization by limiting the mass ratio of ubiquinone to saffron extract to the range of (0.05-0.4):(0.5-3) yields even better results. The optimal effect is achieved when the mass ratio of ubiquinone to saffron extract is 0.3:1.9 (Application Example 10).

[0142] This invention investigated the effect of the mass ratio of menthol lactate in component C on the soothing and redness-improving ability of the composition through application examples 12-14. Application examples 12-14 show that the mass ratio of menthol lactate can increase the a* value improvement rate, thereby affecting the soothing and redness-improving effect of the composition. When the mass ratio of components A, B, and menthol lactate is limited to the range of (0.6-10):(0.11-5.5):(0.001-0.05), the soothing and redness-improving ability of the composition is enhanced. Further optimization by limiting the mass ratio of components A, B, and menthol lactate to the range of (1.5-7):(0.55-3.4):(0.005-0.03) yields an even better composition. The optimal effect is achieved when the mass ratio of components A, B, and menthol lactate is 4:2.2:0.01.

[0143] Therefore, the optimal ratio of each component in the composition of the present invention is: Andrographis paniculata extract: Camellia chrysantha extract: Ubiquinone: Saffron extract: Menthol lactate = 2:2:0.3:1.9:0.01 (Application Example 15).

[0144] This invention investigated the effects of different extraction methods of Camellia chrysantha extract obtained from component A on the composition's ability to lighten dark circles, soothe, and moisturize through Application Examples 15-19. Application Examples 15-19 show that the extraction method of Camellia chrysantha extract has a certain influence on the effects of lightening dark circles, soothing, and moisturizing. The composition obtained using Camellia chrysantha extract-1 showed the best effects in lightening dark circles, soothing, and moisturizing.

[0145] Compared with Application Examples 1-2, the improvement rate of dark circle area was 9-14%, the improvement rate of a* value was less than 6%, and the improvement rate of moisture content was less than 10%. This is because the mass ratio of each component in the composition of Application Examples 1-2 is not within the scope of the present invention, which affects the synergistic effect between components and reduces the efficacy of the composition.

[0146] Compared with Application Examples 3-7, the improvement rate of dark circle area was 10-14%, the improvement rate of a* value was 3-6%, and the improvement rate of moisture content was 3-11%. This is because the composition of Application Examples 3-7 lacked one of the components of the present invention, which affected the synergistic effect between the components and reduced the efficacy of the composition.

[0147] Compared with the comparison application examples 8-12, the improvement rate of dark circle area was 5-10%, the improvement rate of a* value was 2-7%, and the improvement rate of moisture content was 2-11%. This is because the composition of the comparison application examples 8-12 replaced one of the components in the composition of the present invention with other plant extracts, which did not produce a synergistic effect with the specific components selected by the present invention, and also affected the synergistic effect between other components of the present invention, thus reducing the efficacy of the composition.

[0148] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A composition for reducing dark circles under the eyes, characterized in that, The composition comprises component A, component B and component C; Component A consists of Andrographis paniculata extract and Camellia chrysantha extract; Component B consists of ubiquinone and saffron extract; Component C is menthol lactate; The mass ratio of components A, B, and C in the composition is component A: component B: component C = (0.6-10):(0.11-5.5):(0.001-0.05); In component A, the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract is Andrographis paniculata extract: Camellia chrysantha extract = (0.1-5):(0.5-5); In component B, the mass ratio of ubiquinone to saffron extract is ubiquinone:saffron extract = (0.01-0.5):(0.1-5). The preparation method of the Camellia chrysantha extract includes the following steps: After drying the flowers of Camellia chrysantha, the powder was added to an aqueous solution of a eutectic solvent and extracted by ultrasonication. The product was collected, centrifuged, and the supernatant was taken to obtain the Camellia chrysantha extract. The eutectic solvent consisted of choline chloride solution and 1,3-propanediol. The preparation method of the Camellia chrysantha extract meets the following conditions: (a) The mass ratio of choline chloride to 1,3-propanediol is choline chloride: 1,3-propanediol = 1: (1~5). (b) The mass concentration of the eutectic solvent in the aqueous solution of the eutectic solvent is 60-90%; (c) The temperature of the ultrasonic extraction is 40~60℃; the power of the ultrasonic extraction is 120~200W; and the ultrasonic extraction time is 20~50 min. (d) The material-to-liquid ratio for ultrasonic extraction is 1g:(20~40)mL.

2. The composition for lightening dark circles as described in claim 1, characterized in that, The mass ratio of components A, B, and C in the composition is component A: component B: component C = (1.5-7):(0.55-3.4):(0.005-0.03).

3. The composition for lightening dark circles as described in claim 1, characterized in that, In component A, the mass ratio of Andrographis paniculata extract to Camellia chrysantha extract is Andrographis paniculata extract: Camellia chrysantha extract = (0.5-3):(1-4).

4. The composition for lightening dark circles as described in claim 1, characterized in that, In component B, the mass ratio of ubiquinone to saffron extract is ubiquinone:saffron extract = (0.05-0.4):(0.5-3).

5. The use of the composition for reducing dark circles as described in any one of claims 1-4 in the preparation of cosmetics.

6. An eye cream, characterized in that, The eye cream comprises the following components by weight percentage: The composition according to any one of claims 1-4 comprises 1-15% thickener, 0.05-0.5% humectant, 0.5-2% emulsifier, 5-20% preservative, 0.5-3% pH adjuster, and the balance being deionized water.

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