Application of paris polyphylla extract in inhibiting melanin transfer and cosmetics

Abstract

The application discloses application of paris polyphylla extract in inhibition of melanin transfer and cosmetics, and belongs to the technical field of cosmetics. The paris polyphylla extract contains β-ecdysterone, and the β-ecdysterone can inhibit melanin transfer, thereby achieving a whitening effect. The cosmetics containing the paris polyphylla extract have good whitening effect.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, and more specifically, to the application of Paris polyphylla extract in inhibiting melanin transfer and cosmetics. Background Technology

[0002] Human skin color is primarily influenced by pigmentation. Pigmentation not only determines the color of skin, hair, and eyes, but also protects the skin from excessive ultraviolet radiation. The process of pigmentation is very complex, involving the assembly of melanosomes within melanocytes and melanin synthesis. Mature melanosomes are then transferred along the dendritic extension direction towards the distal dendrites. Subsequently, melanosomes detach from melanocytes and enter surrounding keratinocytes, where they are redistributed and degraded. The process of melanosomes moving from the perinuclear region to the dendritic terminals and then to adjacent keratinocytes is called melanosome transport. Studies have shown that melanosome transport plays a crucial role in skin pigmentation.

[0003] Paris polyphylla, a plant belonging to the genus Paris in the family Liliaceae, is also known as seven-leaf Paris or Double-flowered Paris. It gets its name from the fact that its leaves are usually arranged in whorls of seven at the top of the stem, while its flowers are solitary above the whorls of leaves. It is distributed in Sichuan, Yunnan, Guangxi, Guizhou, Jiangxi, Hunan, and other regions of my country. Paris polyphylla has a complex chemical composition. Currently, it is mainly used for anti-tumor, anti-inflammatory, analgesic, platelet aggregation induction, and hemolysis effects. There are no known technologies using it to inhibit melanin transfer.

[0004] In view of this, the present invention is proposed. Summary of the Invention

[0005] One of the objectives of this invention is to provide an application of Paris polyphylla extract in inhibiting melanin transfer.

[0006] The second objective of this invention is to provide a cosmetic containing the above-mentioned Paris polyphylla extract.

[0007] This application can be implemented as follows:

[0008] In a first aspect, this application provides the use of Paris polyphylla extract in inhibiting melanin transfer, wherein the Paris polyphylla extract includes β-ecdysterone.

[0009] In an optional embodiment, Paris polyphylla extract is used to prepare cosmetics with whitening effects.

[0010] In an optional implementation, inhibiting melanin transfer includes at least one of the following aspects:

[0011] a. Reduce the number of melanin dendritic structures;

[0012] b. Shorten the length of melanin dendrites;

[0013] c. Reduce melanin content;

[0014] d. Inhibits melanosome transfer.

[0015] In an optional embodiment, the Paris polyphylla extract is obtained by the following method: ultrasonic-assisted alcohol extraction of Paris polyphylla.

[0016] In an optional embodiment, ultrasound-assisted alcohol extraction includes at least one of the following features:

[0017] Feature 1: Ultrasonic power is 200-500W;

[0018] Feature 2: Ultrasound time is 30-60 minutes;

[0019] Feature 3: The alcohol extraction temperature is 30-60℃;

[0020] Feature 4: The alcohol solution used for alcohol extraction is an ethanol-water solution with an ethanol concentration of 60-85 vt%.

[0021] Feature 5: The ratio of Paris polyphylla medicinal material to alcohol solution is 1:5-1:20.

[0022] In an optional embodiment, the alcohol extract obtained by ultrasound-assisted alcohol extraction is purified; the chromatography column used for purification includes any one of macroporous resin chromatography column and reversed-phase silica gel chromatography column.

[0023] In an optional embodiment, the macroporous resin type used in the macroporous resin chromatography column includes at least one of AB-8, D101, HPD-100, and ADS600.

[0024] Reversed-phase silica gel chromatography columns include the RP-18 reversed-phase silica gel chromatography column or the C18 reversed-phase silica gel chromatography column.

[0025] In an optional embodiment, when purification is performed using a macroporous resin chromatography column, the eluent is an ethanol-water solution; when purification is performed using a reversed-phase silica gel chromatography column, the eluent is a methanol-water solution.

[0026] In an optional implementation, a normal-phase silica gel chromatography column is used for fragmentation before purification using a reversed-phase silica gel chromatography column.

[0027] In an optional embodiment, the normal phase silica gel chromatography column is an 80-100 mesh normal phase silica gel chromatography column.

[0028] In an optional embodiment, the eluent used in the segmentation process is a chloroform-methanol solution.

[0029] In an optional embodiment, prior to purification, the alcohol extract is further subjected to extraction, and the resulting extract is then purified.

[0030] And / or, after purification, the process may also include concentrating the purified eluent.

[0031] Secondly, this application provides a cosmetic product whose ingredients include Paris polyphylla extract, which includes β-ecdysterone.

[0032] The beneficial effects of this application include:

[0033] This application proposes for the first time the application of Paris polyphylla extract containing β-ecdysterone in inhibiting melanocyte transfer. Specifically, β-ecdysterone can exert a whitening effect by inhibiting the ability of melanocytes to transfer. Cosmetics containing β-ecdysterone can correspondingly inhibit tyrosinase activity and melanocyte transfer, thus achieving a whitening effect and showing broad application prospects. Attached Figure Description

[0034] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0035] Figure 1 This is a graph showing the inhibitory effect of each sample on the melanocyte dendritic structure in Test Example 3 of this application;

[0036] Figure 2 This is a graph showing the results of the inhibitory effect of each sample on the number of melanocyte dendrites in Test Example 3 of this application. Detailed Implementation

[0037] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0038] The following is a detailed explanation of the application of the Paris polyphylla extract provided in this application in inhibiting melanin transfer and in cosmetics.

[0039] This application proposes the use of Paris polyphylla extract in inhibiting melanin transfer, wherein the Paris polyphylla extract includes β-ecdysterone.

[0040] The inventors have creatively discovered that Paris polyphylla extract containing β-ecdysterone can effectively inhibit melanocyte transfer. Further research revealed that β-ecdysterone in Paris polyphylla extract can: reduce the number of melanin dendritic structures, and / or shorten the length of melanin dendritic structures, and / or reduce melanin content, and / or inhibit melanosome transfer.

[0041] It should be emphasized that skin whitening effects are not only related to inhibiting melanin production, but also to the ability to inhibit the transfer of melanocytes. In this application, the Paris polyphylla extract achieves its skin whitening effect by inhibiting the transfer of melanocytes.

[0042] For reference, this application proposes that the above-mentioned Paris polyphylla extract can be used to prepare cosmetics with whitening effects, specifically, it can achieve whitening effects by inhibiting melanin transfer.

[0043] In this application, the extract of Paris polyphylla can be obtained by the following method: ultrasonic-assisted alcohol extraction of Paris polyphylla.

[0044] Among them, the medicinal material of Paris polyphylla can be Paris polyphylla from Yunnan, or it can be Paris polyphylla from other producing areas.

[0045] For reference, the ultrasonic power for ultrasonic-assisted alcohol extraction can be 200-500W, such as 200W, 250W, 300W, 350W, 400W, 450W or 500W, or any other value within the range of 200-500W.

[0046] The ultrasonic time for ultrasonic-assisted alcohol extraction can be 30-60 min, such as 30 min, 35 min, 40 min, 45 min, 50 min, 55 min or 60 min, or any other value within the range of 30-60 min.

[0047] The alcohol extraction temperature can be 30-60℃, such as 30℃, 35℃, 40℃, 45℃, 50℃, 55℃ or 60℃, or any other value within the range of 30-60℃.

[0048] The alcohol solution used for alcohol extraction can be an ethanol-water solution with an ethanol concentration of 60-85 vt% (e.g., 60 vt%, 65 vt%, 70 vt%, 75 vt%, 80 vt%, or 85 vt%). Furthermore, the use of ethanol-water solutions of other volume concentrations is not excluded, nor is the use of other organic reagents such as methanol for alcohol extraction.

[0049] The ratio of Paris polyphylla medicinal material to alcohol solution can be 1:5-1:20, such as 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19 or 1:20, or any other value within the range of 1:5-1:20.

[0050] In some implementations, the Paris polyphylla herb may be first soaked in an alcohol solution, and then ultrasonic-assisted extraction may be performed.

[0051] Furthermore, the alcohol extract obtained by ultrasound-assisted alcohol extraction is purified.

[0052] In some embodiments, the alcohol extract can be centrifuged first, and the supernatant can be collected to remove impurities. The centrifugation speed can be 4500-5500 rpm, such as 4500 rpm, 4800 rpm, 5000 rpm, 5200 rpm, or 5500 rpm.

[0053] The obtained supernatant is extracted to obtain an extract, which is then purified. For example, the extractant used can be n-butanol, etc. The extraction can be performed once, twice, three times, or more. When the number of extractions is ≥2, the extracts obtained from each extraction are combined.

[0054] Alternatively, the obtained supernatant can be directly subjected to vacuum distillation until the alcohol in the alcohol extract is removed, followed by purification.

[0055] In this application, the chromatography column used for purification may include, for example, any one of a macroporous resin chromatography column and a reversed-phase silica gel chromatography column. The macroporous resin type used in the macroporous resin chromatography column includes at least one of AB-8, D101, HPD-100, and ADS600. The reversed-phase silica gel chromatography column includes an RP-18 reversed-phase silica gel chromatography column or a C18 reversed-phase silica gel chromatography column.

[0056] When purification is performed using a macroporous resin chromatography column, the eluent is an ethanol-water solution. In some embodiments, gradient elution is performed with ethanol in the eluent at successively 0 vt%, 30 vt%, 70 vt%, and 90 vt%, and the volume of eluent used for each elution gradient may, for example, but is not limited to, two beds. The eluent obtained from the 70 vt% ethanol-water solution is used for subsequent operations. In other embodiments, the elution conditions can be adjusted as appropriate.

[0057] When purification is performed using a reversed-phase silica gel chromatography column, the eluent is a methanol-water solution. In some embodiments, gradient elution is performed with methanol to water volume ratios of 10:90, 20:80, 30:70, 40:60, and 50:50, where each elution gradient corresponds to, for example, but not limited to, two bed layers of eluent. The eluent obtained from the 50:50 methanol-water ratio is used for subsequent operations. In other embodiments, the elution conditions can be adjusted as appropriate.

[0058] In some embodiments, before purification using a reversed-phase silica gel column, a normal-phase silica gel column can be used for fragmentation. For example, an 80-100 mesh normal-phase silica gel column can be used for fragmentation, and the eluent used in the fragmentation process can be, for example, a chloroform-methanol solution. More specifically, in some embodiments, fragmentation can be performed as follows: gradient elution with chloroform to methanol at volume ratios of 5:1, 4:1, 3:1, 2:1, 1:1, and 0:1, where the volume of eluent used for each elution gradient can be, for example, but is not limited to, two bed layers. The eluent obtained at a chloroform to methanol volume ratio of 5:1 is then used for subsequent reversed-phase silica gel column purification. In other embodiments, the elution conditions can be adjusted appropriately.

[0059] After purification, the eluent obtained from the purification process is concentrated.

[0060] For example, concentration can be carried out by vacuum distillation.

[0061] As mentioned above, the Paris polyphylla extract obtained by the preparation method provided in this application contains a high content of β-ecdysterone, which can effectively inhibit the activity of tyrosinase and inhibit the transfer of melanocytes, thus having a highly effective whitening effect.

[0062] It should be noted that the Paris polyphylla extract in this application is not limited to the method described above; any method that can extract Paris polyphylla extract containing β-ecdysterone is acceptable.

[0063] In addition, this application also provides a cosmetic product whose ingredients include Paris polyphylla extract, which includes β-ecdysterone.

[0064] The effects and preparation methods of Paris polyphylla extract can be found in the aforementioned content, and will not be elaborated upon here.

[0065] For reference, the dosage forms of cosmetics in this application may include, for example, creams, lotions, liquids, gels, oils, powders, sheets, films, freeze-dried products, block powders or solids, clays, aerosols, organic solvents, wax-based products, and any other applicable dosage forms of cosmetics that act directly or indirectly on the skin. For example, they may include, for instance, whitening essences, moisturizing essences, sunscreen essences, anti-wrinkle essences, toners, lotions, makeup removers, cleansing milks, moisturizing creams, sunscreens, sunblocks, sunscreen lotions, body lotions, cold creams, moisturizing creams, day creams, night creams, moisturizing gels, face masks, facial cleansers, facial foams, facial washes, foundations, hand creams, shower gels, cosmetic soaps, bar soaps, setting sprays, or moisturizing sprays, etc.

[0066] Besides Paris polyphylla extract, cosmetic ingredients can also include any functional ingredients and excipients available in the cosmetic field, which will not be listed here.

[0067] For reference, the content of β-ecdysterone in cosmetics can be 0.0000005-0.01%, preferably 0.0000005-0.001%.

[0068] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0069] Example 1

[0070] This embodiment provides a method for preparing Paris polyphylla extract, as detailed below:

[0071] The raw material of Paris polyphylla purchased from Yunnan Xicao was dried in a forced-air drying oven at 55°C and then pulverized into 10-mesh powder using a traditional Chinese medicine pulverizer.

[0072] Take the above powder and use 85% ethanol-water solution as the extraction solvent to soak for 1 hour, with a material-liquid ratio of 1:10. Set the extraction temperature to 50℃, the ultrasonic power to 500W, and the ultrasonic time to 50min for ultrasonic extraction. Centrifuge at 5000rpm for 10min, take the supernatant (i.e., ethanol extract), and filter to remove impurities.

[0073] The alcohol extract was extracted twice with n-butanol, and the extracts were combined.

[0074] The extract was divided into segments using an 80-mesh normal-phase silica gel column and then eluted with a chloroform-methanol solution at volume ratios of 5:1, 4:1, 3:1, 2:1, 1:1, and 0:1. Each elution gradient corresponded to two bed volumes of eluent.

[0075] The eluent obtained from the 5:1 (chloroform:methanol) elution was purified using a C18 reversed-phase silica gel column. Gradient elution was then performed with methanol-water solution at methanol:water ratios of 10:90, 20:80, 30:70, 40:60, and 50:50 (methanol:water volume ratios), with each gradient corresponding to two bed volumes of eluent. 200 mL of the 50:50 (methanol:water) elution eluent was evaporated to dryness to obtain the Paris polyphylla extract powder.

[0076] Example 2

[0077] This embodiment provides a method for preparing Paris polyphylla extract, as detailed below:

[0078] The raw material of Paris polyphylla purchased from Xicao, Yunnan was made into powder, and the steps were the same as in Example 1.

[0079] Take the above powder and use 70 wt% ethanol-water solution as the extraction solvent, with a material-to-liquid ratio of 1:10. Set the extraction temperature to 50℃, the ultrasonic power to 200W, and the ultrasonic time to 30 min for ultrasonic extraction. Centrifuge and filter at 8000 rpm for 2 min and take the supernatant (i.e., ethanol extract). Take the filter residue and repeat the above steps once. Mix the filtrate and filter to remove impurities.

[0080] The alcohol extract was distilled under reduced pressure until no alcohol odor was detected. It was then purified using AB-8 macroporous resin. Gradient elution was performed with ethanol in the eluent (ethanol-water solution) at volume percentages of 0 vt%, 30 vt%, 70 vt%, and 90 vt%, respectively. Each elution gradient corresponded to two bed volumes of eluent.

[0081] Take 200 mL of the eluent obtained by washing with 70 VT% ethanol-water, evaporate to dryness to obtain the 70 ethanol fraction of Paris polyphylla extract powder.

[0082] Example 3

[0083] This embodiment provides a method for preparing Paris polyphylla extract, as detailed below:

[0084] The raw material of Paris polyphylla purchased from Xicao, Yunnan was made into powder, and the steps were the same as in Example 1.

[0085] Take the above powder and use 70 wt% ethanol-water solution as the extraction solvent, with a material-to-liquid ratio of 1:20. Set the extraction temperature to 60℃, the ultrasonic power to 500W, and the ultrasonic time to 50min for ultrasonic extraction. Centrifuge and filter at 8000rpm for 2min and take the supernatant (i.e., ethanol extract). Take the filter residue and repeat the above steps once. Mix the filtrate and filter to remove impurities.

[0086] The alcohol extract was distilled under reduced pressure until no alcohol odor was detected. It was then purified using AB-8 macroporous resin. Gradient elution was performed with ethanol in the eluent (ethanol-water solution) at volume percentages of 0 vt%, 30 vt%, 70 vt%, and 90 vt%, respectively. Each elution gradient corresponded to two bed volumes of eluent.

[0087] Take 200 mL of the eluent obtained by washing with 70 VT% ethanol-water, evaporate to dryness to obtain the 70 ethanol fraction of Paris polyphylla extract powder.

[0088] Example 4

[0089] This embodiment provides a method for preparing Paris polyphylla extract, as detailed below:

[0090] The raw material of Paris polyphylla purchased from Xicao, Yunnan was made into powder, and the steps were the same as in Example 1.

[0091] Take the above powder and use 60 wt% ethanol-water solution as the extraction solvent, with a material-liquid ratio of 1:5. Set the extraction temperature to 30℃, the ultrasonic power to 200W, and the ultrasonic time to 30min for ultrasonic extraction. Centrifuge and filter at 8000rpm for 2min and take the supernatant (i.e., ethanol extract). Take the filter residue and repeat the above steps once. Mix the filtrate and filter to remove impurities.

[0092] The alcohol extract was distilled under reduced pressure until no alcohol odor was detected. It was then purified using AB-8 macroporous resin. Gradient elution was performed with ethanol in the eluent (ethanol-water solution) at volume percentages of 0 vt%, 30 vt%, 70 vt%, and 90 vt%, respectively. Each elution gradient corresponded to two bed volumes of eluent.

[0093] Take 200 mL of the eluent obtained by washing with 70 VT% ethanol-water, evaporate to dryness to obtain the 70 ethanol fraction of Paris polyphylla extract powder.

[0094] Test Example 1

[0095] Determination of active ingredients in extracts

[0096] Using the Paris polyphylla extracts obtained in Examples 1-2 as test samples, the β-ecdysterone content in the Paris polyphylla extracts was determined by high performance liquid chromatography, and the results are shown in Table 1.

[0097] The determination methods and conditions are as follows:

[0098] Each sample was prepared into a 4 mg / mL methanol solution;

[0099] One vial of β-ecdysterone (98% purity, purchased from the National Institutes for Food and Drug Control) standard was prepared into a stock solution with a mass concentration of 1 mg / mL using 95% ethanol. The stock solution was then diluted with purified water to prepare a series of β-ecdysterone solutions with concentrations of 0.10 mg / mL, 0.15 mg / mL, 0.20 mg / mL, 0.25 mg / mL, and 0.30 mg / mL. These solutions were then filtered through a 0.22 μm needle membrane for later use.

[0100] Chromatographic column: Agilent Extend-C18 ODS column (4.6×150mm, particle size 5μm);

[0101] Mobile phase: Acetonitrile: Methanol: Water = 1:3:4;

[0102] Column temperature: 30℃;

[0103] Detection wavelength: 243nm;

[0104] Flow rate: 1.00 mL / min;

[0105] Injection volume: 10 μL.

[0106] Table 1. β-ecdysterone content

[0107] sample β-ecdysterone (%) Example 1 34.7 Example 2 2.5

[0108] As can be seen from Table 1, the Paris polyphylla extracts prepared in Examples 1-2 all contain β-ecdysterone, and the β-ecdysterone content of the Paris polyphylla extract prepared in Example 1 exceeds 30 wt%.

[0109] Test Example 2

[0110] B16 tyrosinase activity inhibition test

[0111] Taking β-ecdysterone (98% purity, purchased from Huzhou Aino Company) as an example, it was diluted with ultrapure water to prepare samples A, B, and C. The concentration of sample A was 0.2 ppm, the concentration of sample B was 1 ppm, and the concentration of sample C was 5 ppm. Taking the Paris polyphylla extract containing 34.7% β-ecdysterone prepared in Example 1 as an example, it was diluted with ultrapure water to prepare sample D with a concentration of 5 ppm. α-Arbutin was used as a positive control and diluted with ultrapure water to a concentration of 500 μmol / L.

[0112] B16 cells in the logarithmic growth phase were seeded in 96-well plates at a certain number of cells and incubated in a CO2 incubator at 37°C and 5% CO2 concentration until they reached 70% confluence. Starvation medium was then added overnight. The next day, the above-mentioned sample was added, and the cells were incubated for another 24 hours. A solvent control group was also set up. After sample processing, the cells were washed twice with PBS, and tyrosinase activity detection solution was added. The cells were then incubated at 37°C for 0.5 hours, and the absorbance (OD value) at 490 nm was measured. The results were calculated and analyzed. The results showed that none of the tested samples were toxic. The inhibition rates of B16 tyrosinase activity in each group are shown in Table 2.

[0113] Table 2B16 Tyrosinase Activity Inhibition Rate

[0114] sample Tyrosinase activity inhibition rate (%) A 2.55 B 7.31 C 11.89 D 1.18 α-Arbutin 16.17 NT (solvent control) 0

[0115] As shown in Table 2, even low concentrations of β-ecdysterone have a strong tyrosinase activity inhibition rate, and the Paris polyphylla extract prepared in Example 1 containing 34.7% also has a tyrosinase activity inhibition rate.

[0116] Test Example 3

[0117] Tyrosinase activity inhibition test

[0118] The Paris polyphylla extract containing 34.7% β-ecdysterone prepared in Example 1 and the Paris polyphylla extract containing 2.5% β-ecdysterone prepared in Example 2 were used as test samples. All test samples were diluted with methanol to 0.1 wt%.

[0119] Using 100 μg / mL ascorbic acid (Vc) as a positive control, DOPA solution (1 mg / mL), phosphate buffer (pH = 6.8), and the test sample were extracted and preheated at 37℃ for 10 min. Tyrosinase (106 U / mL) was preheated for 2 min. 1 mL of sample solution and 0.5 mL of tyrosinase solution were transferred to a T-group test tube, 1 mL of sample solution and 0.5 mL of phosphate buffer were transferred to a T0-group test tube, 1 mL of phosphate buffer and 0.5 mL of tyrosinase solution were transferred to a C-group test tube, and 1.5 mL of phosphate buffer was transferred to a C0-group test tube. All test tubes were preheated at 37℃ for 2 min. 2 mL of DOPA solution was transferred to all reaction tubes. After reacting at 37℃ for 10 min, the absorbance at 517 nm was measured.

[0120] The clearance rate is calculated using the formula below. The higher the clearance rate, the stronger the whitening effect. The results are shown in Table 3.

[0121] Sweep rate I (%) = [1 - (T - T0) / (C - C0)] × 100;

[0122] In the formula, T represents the absorbance of 1 mL sample solution plus 0.5 mL tyrosinase solution plus 2 mL dopa solution; T0 represents the absorbance of 1 mL sample solution plus 0.5 mL phosphate buffer plus 2 mL dopa solution; C0 represents the absorbance of 1.5 mL phosphate buffer plus 2 mL dopa solution; and C represents the absorbance of 1 mL phosphate buffer plus 0.5 mL phosphate buffer plus 2 mL dopa solution.

[0123] Table 3 Tyrosinase activity inhibition rate

[0124] sample Tyrosinase activity inhibition rate (%) Example 1 38.30 Example 2 29.03 Vc 28.73

[0125] As shown in Table 3, at a test sample concentration of 0.1 wt%, the Paris polyphylla extracts containing β-ecdysterone obtained in Examples 1-2 all exhibited higher tyrosinase activity inhibition rates than vitamin C, indicating that the Paris polyphylla extract has a strong whitening effect.

[0126] Test Example 4

[0127] Melanocyte dendrite test

[0128] Taking the Paris polyphylla extract containing 34.7% β-ecdysterone prepared in Example 1 as an example, it was diluted with DMSO to form Sample 1, Sample 2, Sample 3 and Sample 4, wherein the concentration of Sample 1 was 0.0005 wt%, the concentration of Sample 2 was 0.0001 wt%, the concentration of Sample 3 was 0.00005 wt%, and the concentration of Sample 4 was 0.00001 wt%.

[0129] A mixture of revived primary human keratinocytes and primary human melanocytes was seeded into wells at a specific cell count and incubated at 37°C in a 5% CO2 incubator. When the cells reached 60%–70% confluence, they were treated with the above-mentioned sample and incubated for another 24 hours. After exposure, the active agent was discarded, and the cells were fixed with 4% paraformaldehyde, stained with phosphate-dopa staining solution, and photographed under the same exposure. The dendritic structure (number of melanocyte dendrites / number of melanocytes) was statistically analyzed. The results are shown below. Figure 1 and Figure 2 As shown.

[0130] Depend on Figure 1 and Figure 2 The results showed that, compared with the control group with added DMSO, the extracts of Paris polyphylla at concentrations of 0.0005wt%, 0.0001wt%, 0.00005wt%, and 0.00001wt% all significantly inhibited melanocyte growth, and the melanin content in melanocytes was reduced in all cases, as observed by staining. At the same time, it was observed that the dendritic structures of melanocytes were shrunken under the above four concentrations, and the number of dendritic structures was significantly reduced, indicating that the extract of Paris polyphylla has a significant inhibitory effect on melanocyte transfer.

[0131] In summary, this application proposes the application of Paris polyphylla extract containing β-ecdysterone in inhibiting melanocyte transfer. β-ecdysterone can exert a whitening effect by inhibiting the ability of melanocytes to transfer. Cosmetics containing β-ecdysterone correspondingly have the effect of inhibiting tyrosinase activity and inhibiting melanocyte transfer, thus achieving a whitening effect.

[0132] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. The application of a Paris polyphylla extract in the preparation of products with whitening effects, wherein, The Paris polyphylla extract includes β-ecdysterone; The Paris polyphylla extract was obtained by the following method: Yunnan Paris polyphylla raw material purchased from Yunnan Xicao was placed in a forced-air drying oven and dried at 55°C, and then pulverized with a traditional Chinese medicine pulverizer to make 10-mesh powder. Take the above powder and use 85 wt% ethanol-water solution as the extraction solvent to soak for 1 hour, with a material-to-liquid ratio of 1:

10. Set the extraction temperature to 50℃, the ultrasonic power to 500W, and the ultrasonic time to 50 min for ultrasonic extraction. Centrifuge at 5000 rpm for 10 min, take the supernatant, and filter to remove impurities. The alcohol extract was extracted twice with n-butanol, and the extracts were combined. The extract was divided into segments using an 80-mesh normal-phase silica gel column and then eluted with a chloroform-methanol solution at volume ratios of 5:1, 4:1, 3:1, 2:1, 1:1 and 0:

1. Each elution gradient corresponded to two bed volumes of eluent. The eluent obtained from the chloroform-methanol mixture at a volume ratio of 5:1 was purified using a C18 reversed-phase silica gel column. Gradient elution was then performed using methanol-water solution at methanol-water volume ratios of 10:90, 20:80, 30:70, 40:60, and 50:50, with each elution gradient corresponding to two bed volumes of eluent. 200 mL of the eluent obtained from the 50:50 methanol-water mixture was taken and evaporated to dryness to obtain the Paris polyphylla extract powder.

2. The application according to claim 1, characterized in that, The Paris polyphylla extract is used to prepare cosmetics with whitening effects.

3. The application according to claim 1 or 2, characterized in that, The Paris polyphylla extract is used to inhibit melanin transfer; inhibition of melanin transfer includes at least one of the following aspects: a. Reduce the number of melanin dendritic structures; b. Shorten the length of melanin dendrites; c. Reduce melanin content; d. Inhibits melanosome transfer.

4. The application according to claim 1, characterized in that, After purification, the process also includes concentrating the purified eluent.

Images