Use of a compound containing dendrobium officinale leaf extract in the preparation of a whitening and freckle-removing preparation

Abstract

The application discloses application of a complex containing Dendrobium officinale leaf extract in preparation of a whitening and freckle-removing preparation, and the complex comprises the Dendrobium officinale leaf extract, a Rhizoma Polygonati extract and beta arbutin. After the components of the complex are mixed in a specific ratio and fully combined and synergized, the ability of inhibiting tyrosinase activity can be significantly improved by 300%, 175% and 87% respectively compared with the Dendrobium officinale leaf, the Rhizoma Polygonati and the beta arbutin, the melanin generation in zebra fish within 72 hours can be effectively inhibited, the melanin content can be obviously reduced, the melanin inhibition rate is increased by 394%, 532%, 24.14% and 37.39% respectively compared with the Dendrobium officinale leaf group, the Rhizoma Polygonati group, the beta arbutin group and the Dendrobium officinale leaf-Rhizoma Polygonati group, the Dendrobium officinale leaf-Rhizoma Polygonati complex can obviously improve the melanin inhibition rate in zebra fish, the combination of the two and the beta arbutin can make the melanin inhibition rate reach 98.70%, and the whitening and freckle-removing effect of skin can be greatly enhanced.

Description

Technical Field

[0001] This invention belongs to the field of Dendrobium officinale application technology, specifically relating to the application of a complex containing Dendrobium officinale leaf extract in the preparation of whitening and freckle-removing preparations. Background Technology

[0002] Due to various internal and external factors, such as genetic factors, ultraviolet radiation, and hormonal changes, people's skin often experiences pigmentation and dullness, losing its original radiance and affecting appearance, self-confidence, and quality of life. The main factor influencing skin color is the content and distribution of melanin. Melanin is produced by the oxidation of tyrosine under the action of tyrosinase. Therefore, inhibiting tyrosinase, the key rate-limiting enzyme in the melanin production pathway, is one of the main mechanisms for whitening and removing blemishes.

[0003] In recent years, researchers have gradually shifted their focus from chemically synthesized products to safer and more effective natural plant extracts in their search for ingredients with whitening and blemish-removing effects. Dendrobium officinale Kiumra et Migo, a plant belonging to the genus Dendrobium in the Orchidaceae family, is used medicinally for its stems. However, the leaves, being a non-medicinal part, are produced in huge quantities annually, and their utilization is often insufficient during harvesting and use, resulting in significant resource waste.

[0004] Polygonatum is the dried rhizome of Polygonatum sibiricum Red., Polygonatum kingianum Coll. et Hemsl., or Polygonatum cyrtonema Hua., all belonging to the Liliaceae family. It has various pharmacological effects and has been widely used in the medical and food fields. However, research on the whitening and freckle-removing effects of Polygonatum extract is scarce, indicating significant potential for development.

[0005] Beta-arbutin is a commonly used skin-whitening ingredient, but it can degrade into hydroquinone due to environmental factors and dosage. Long-term excessive use can lead to dry skin, redness, and white patches, thus affecting human health. Patent CN202310864944.2 discloses a cosmetic composition containing arbutin with skin-whitening effects and its preparation method, and measures the in vitro tyrosinase activity inhibition rate. However, the tyrosinase activity inhibition rate in its three embodiments is only 50%–54%.

[0006] Currently, there is a lack of methods in this field for the preparation and application of complexes of Dendrobium officinale leaves, Polygonatum sibiricum extract and β-arbutin. Summary of the Invention

[0007] To address the above-mentioned problems, the present invention aims to provide an application of a complex containing Dendrobium officinale leaf extract in the preparation of whitening and freckle-removing preparations.

[0008] The specific technical solution is as follows:

[0009] The application of a complex containing Dendrobium officinale leaf extract in the preparation of a whitening and freckle-removing preparation, the complex comprising Dendrobium officinale leaf extract, Polygonatum sibiricum extract and β-arbutin, wherein the mass ratio of Dendrobium officinale leaf extract to Polygonatum sibiricum extract is 1:0.04-22.60, and the mass ratio of the total mass of Dendrobium officinale leaf extract and Polygonatum sibiricum extract to β-arbutin is 0.6-15.2:1.

[0010] Furthermore, the mass ratio of Dendrobium officinale leaf extract to Polygonatum sibiricum extract was 1:0.26-13.80, and the mass ratio of the total mass of Dendrobium officinale leaf extract and Polygonatum sibiricum extract to β-arbutin was 0.18-25.50:1.

[0011] Furthermore, the mass ratio of Dendrobium officinale leaf extract, Polygonatum sibiricum extract, and β-arbutin was 25:78:10.

[0012] Furthermore, the preparation method of Dendrobium officinale leaf extract is as follows: Dendrobium officinale leaves are dried by forced air at 200℃, crushed by a pulverizer, soaked in a solvent (1%–80% ethanol aqueous solution) for 0.2–20 h, extracted by reflux at 30–100℃ for 1–10 h, and extracted 1–15 times. After filtration, the filtrate is concentrated under reduced pressure at 20–70℃, and the concentrate is freeze-dried to obtain Dendrobium officinale leaf extract.

[0013] Furthermore, the preparation method of Polygonatum extract is as follows: Polygonatum is dried by forced air at 200℃, crushed by a pulverizer, soaked in a solvent (5%–75% ethanol aqueous solution) for 0.5–18 hours, refluxed for extraction at 60–100℃ for 2–8 hours, and extracted 1–10 times. After filtration, the filtrate is concentrated under reduced pressure at 20–80℃, and the concentrate is freeze-dried to obtain Polygonatum extract.

[0014] The beneficial effects of this invention are as follows:

[0015] 1) Dendrobium officinale leaf extract is rich in polyphenols, flavonoids, polysaccharides, and amino acids, and has a good free radical scavenging ability. However, when used alone, its ability to inhibit tyrosinase activity is weak, and its whitening and spot-removing effects are not obvious. Polygonatum contains polysaccharides, saponins, flavonoids, and other chemical components, but its extract has a dark color, which affects its aesthetics. Existing decolorization methods will affect its whitening, spot-removing, and antioxidant effects. β-arbutin has a strong whitening effect, but it is irritating. Long-term and excessive use can cause dry skin, redness, and swelling. In severe cases, it can be damaged by the degradation product hydroquinone, resulting in white spots or cancer. Therefore, when using this ingredient, attention must be paid to its dosage, and the lowest amount should be used to achieve the best effect. 2) After the three ingredients are mixed in a specific ratio, they are fully organically combined and synergistically enhanced. Compared with their individual applications, their ability to inhibit tyrosinase activity is greatly improved. Through in vitro tyrosinase activity inhibition experiments and zebrafish whitening model verification, the whitening and spot-removing ability of Dendrobium officinale leaf extract, Polygonatum sibiricum extract, and β-arbutin complex at a specific ratio far exceeds that of single or two traditional Chinese medicine complexes and β-arbutin under the same total dosage conditions.

[0016] 3) While increasing safety and reducing product costs, this invention fully develops the non-medicinal parts of Dendrobium officinale, avoids resource waste, and has good prospects for market development and application. Attached Figure Description

[0017] Figure 1 The effect of different treatment groups on the inhibition of melanin on the surface of zebrafish embryos. Detailed Implementation

[0018] The present invention will be further described below with reference to the accompanying drawings and embodiments. However, the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. The present invention covers any substitutions, modifications, equivalent methods, and solutions made within the scope of the invention as defined in the claims. Where specific techniques or conditions are not specified in the embodiments, they shall be performed in accordance with the techniques or conditions described in the literature in the art or in accordance with the product manual. Materials or equipment whose manufacturers are not specified can be obtained through conventional channels.

[0019] Example 1

[0020] Preparation of Dendrobium officinale leaf extract: Dendrobium officinale leaves were dried in a forced-air oven at 200℃, crushed with a pulverizer, and then soaked in 5 times the volume of 35% ethanol-water for 1 hour. After reflux extraction, the extraction temperature was 80℃, and the solution was kept at a gentle boil for 8.5 hours. The extraction was repeated 10 times. After cooling and filtration, the filtrate was concentrated and freeze-dried at 70℃. The concentrated solution was pre-frozen and then freeze-dried in a freeze dryer to obtain Dendrobium officinale leaf freeze-dried powder extract.

[0021] Preparation Example 1: Preparation of 0.001 mg / ml Dendrobium officinale leaf extract: Take 0.01 mg of the Dendrobium officinale leaf extract prepared above and dissolve it in 10 ml of 35% ethanol aqueous solution until fully dissolved;

[0022] Preparation Example 2: Preparation of 0.01 mg / ml Dendrobium officinale leaf extract: Take 0.1 mg of the Dendrobium officinale leaf extract prepared above and dissolve it in 10 ml of 35% ethanol aqueous solution until fully dissolved;

[0023] Preparation Example 3: Preparation of 0.1 mg / ml Dendrobium officinale leaf extract: Take 1 mg of the Dendrobium officinale leaf extract prepared above and dissolve it in 10 ml of 35% ethanol aqueous solution until fully dissolved;

[0024] Preparation Example 4: Preparation of 1 mg / ml Dendrobium officinale leaf extract: Take 10 mg of the Dendrobium officinale leaf extract prepared above, dissolve it in 10 ml of 35% ethanol aqueous solution, and dissolve it completely;

[0025] Preparation Example 5: Preparation of 10 mg / ml Dendrobium officinale leaf extract: Take 100 mg of the Dendrobium officinale leaf extract prepared above and dissolve it in 10 ml of 35% ethanol aqueous solution until fully dissolved.

[0026] Example 2

[0027] Preparation of Polygonatum extract:

[0028] The Polygonatum was dried at 200℃ by forced air drying, crushed by a pulverizer, and then soaked in 2-25 times its volume of 10%-65% ethanol-water for 2 hours. After reflux extraction, the extraction temperature was 90℃, and the solution was kept at a gentle boil for 7.5 hours. The extraction was repeated 7 times. After cooling and filtration, the filtrate was concentrated by rotary evaporation at 78℃. The concentrated solution was pre-frozen and then placed in a freeze dryer to obtain the freeze-dried Polygonatum extract.

[0029] Preparation Example 6: Preparation of 0.001 mg / ml Polygonatum extract: Take 0.01 mg of the Dendrobium officinale leaf extract prepared above, dissolve it in 10 ml of 35% ethanol aqueous solution, and dissolve it completely;

[0030] Preparation Example 7: Preparation of 0.01 mg / ml Polygonatum extract: Take 0.1 mg of the Dendrobium officinale leaf extract prepared above, dissolve it in 10 ml of 35% ethanol aqueous solution, and dissolve it completely;

[0031] Preparation Example 8: Preparation of 0.1 mg / ml Polygonatum extract: Take 1 mg of the Dendrobium officinale leaf extract prepared above, dissolve it in 10 ml of 35% ethanol aqueous solution, and dissolve it completely;

[0032] Preparation Example 9: Preparation of 1 mg / ml Polygonatum extract: Take 10 mg of Dendrobium officinale leaf extract prepared above, dissolve it in 10 ml of 35% ethanol aqueous solution, and dissolve it completely;

[0033] Preparation Example 10: Preparation of 10 mg / ml Polygonatum extract: Take 100 mg of the Dendrobium officinale leaf extract prepared above and dissolve it in 10 ml of 35% ethanol aqueous solution until fully dissolved.

[0034] Example 3

[0035] Preparation of complexes of Dendrobium officinale leaf extract and Polygonatum sibiricum extract in different proportions

[0036] Complexes were prepared by mixing Dendrobium officinale leaf extract solutions and Polygonatum sibiricum extract solutions of different concentrations obtained in Examples 1 and 2. The mass ratios of Dendrobium officinale leaf extract and Polygonatum sibiricum extract were 13:87 (Preparation Example 11), 20:80 (Preparation Example 12), 25:85 (Preparation Example 13), 40:60 (Preparation Example 14), 55:45 (Preparation Example 15), 48:52 (Preparation Example 16), 68:32 (Preparation Example 17), 88:12 (Preparation Example 18), and 95:5 (Preparation Example 19).

[0037] Example 4

[0038] Samples of Dendrobium officinale leaf extract, Polygonatum sibiricum extract, β-arbutin, Dendrobium officinale leaf-Polygonatum sibiricum extract complex, and Dendrobium officinale leaf extract, Polygonatum sibiricum extract, and β-arbutin complex at the same concentration were prepared, as well as samples of Dendrobium officinale leaf extract, Polygonatum sibiricum extract, and β-arbutin complex at different ratios.

[0039] Preparation Example 20: Preparation of Dendrobium officinale leaf extract sample: Take 0.015 mg of Dendrobium officinale leaf extract from Example 1, add 10 ml of 35% ethanol aqueous solution to dissolve, and prepare the target sample.

[0040] Preparation Example 21: Preparation of Polygonatum extract sample: Take 0.015 mg of Polygonatum extract from Example 2, add 10 ml of 35% ethanol aqueous solution to dissolve, and prepare the target sample.

[0041] Preparation Example 22: Preparation of β-arbutin solution: Take 0.015 mg of β-arbutin, add 10 ml of 35% ethanol aqueous solution to dissolve it, and prepare the target sample.

[0042] Preparation Example 23: Preparation of a complex sample of Dendrobium officinale leaf and Polygonatum sibiricum extract: Take 0.015 mg of a mixture of Dendrobium officinale leaf and Polygonatum sibiricum extract in a ratio of 68:32, add 10 ml of 35% ethanol aqueous solution to dissolve, and prepare the target sample.

[0043] Preparation of Dendrobium officinale leaf, Polygonatum sibiricum and β-arbutin complex: Take Dendrobium officinale leaf extract solution, Polygonatum sibiricum extract solution and β-arbutin solution, mix them, and make Dendrobium officinale leaf extract, Polygonatum sibiricum extract and β-arbutin in the following ratios: 30:60:45 (Preparation Example 24), 25:78:10 (Preparation Example 25), 10:57:5 (Preparation Example 26), 10:57:89 (Preparation Example 27) and 5:45:80 (Preparation Example 28).

[0044] Example 5

[0045] Samples of blank and the same concentration of Dendrobium officinale leaf, Polygonatum sibiricum, β-arbutin, Dendrobium officinale leaf-Polygonatum sibiricum complex, and Dendrobium officinale leaf-Polygonatum sibiricum-β-arbutin complex were prepared for subsequent evaluation of melanin content in zebrafish models.

[0046] Preparation Example 29: Without adding extract, add 100 ml of zebrafish culture medium;

[0047] Preparation Example 30: 0.001 mg of Dendrobium officinale leaf extract lyophilized powder was dissolved in 100 ml of zebrafish culture medium to obtain the target sample;

[0048] Preparation Example 31: 0.001 mg of lyophilized Polygonatum extract powder was dissolved in 100 ml of zebrafish culture medium to obtain the target sample;

[0049] Preparation Example 32: Dissolve 0.001 mg of β-arbutin in 100 ml of zebrafish culture medium to obtain the target sample;

[0050] Preparation Example 33: Take 0.001 mg of freeze-dried powder of Dendrobium officinale leaf-Polygonatum extract in a ratio of 68:32, dissolve it in 100 ml of zebrafish culture medium to obtain the target sample;

[0051] Preparation Example 34: Take 0.001 mg of Dendrobium officinale leaf-Polygonatum sibiricum-β-arbutin in a ratio of 25:78:10 and dissolve it in 100 ml of zebrafish culture medium to obtain the target sample;

[0052] Example 6

[0053] Tyrosinase activity inhibition experiment

[0054] Experimental Method: Using a pipette, accurately pipette the four reaction solutions according to the volumes in the table below into 96-well plates. The inhibitors were different concentrations of Polygonatum sibiricum, Dendrobium officinale leaf extract, β-arbutin, Dendrobium officinale leaf-Polygonatum sibiricum compound, and Dendrobium officinale leaf-Polygonatum sibiricum-β-arbutin. Add 80 μL of L-tyrosine solution, vortex to mix, and incubate at 37℃ for 10 min. Then, add 50 μL of tyrosinase to both the sample tube and the enzyme reaction tube. After reacting for 10 min, immediately measure the reaction using a microplate reader. A 样 A 样品本底 A酶反应管 A 溶剂本底 The absorbances at a wavelength of 475 nm are A1, A0, B1, and B0, respectively. The inhibition rate of the sample on tyrosinase activity is calculated using formula (1-1):

[0055] Inhibition rate (%) = [1 - (A1 - A0) / (B1 - B0)] × 100%

[0056] Table 1. Sample dosage for determining the inhibition rate of tyrosinase activity.

[0057]

[0058] Table 2. Inhibition rate of Dendrobium officinale leaf extract on tyrosinase activity

[0059] Concentration (mg / mL) Inhibition rate / (%) 0.001 23.25±5.15 0.01 31.83±1.65 0.1 37.21±1.86 1 46.18±5.66 10 63.66±4.31

[0060] Table 3. Inhibition rate of Polygonatum extract on tyrosinase activity

[0061] Concentration (mg / mL) Inhibition rate / (%) 0.001 35.35±4.95 0.01 44.11±1.50 0.1 56.45±4.26 1 68.56±2.76 10 81.53±3.50

[0062] As shown in Table 2, the inhibitory effect of Dendrobium officinale leaf extract on tyrosinase activity increases with increasing concentration, exhibiting a dose-dependent effect. However, at high concentrations, the inhibitory effect on tyrosinase activity only reaches about 64%, indicating a less significant inhibitory effect. As shown in Table 3, the inhibitory effect of Polygonatum sibiricum extract on tyrosinase activity is dose-dependent, with the inhibitory effect increasing with increasing concentration. However, excessively high concentrations can lead to a darker solution color, affecting the appearance of the subsequent products. Therefore, it is necessary to reduce the dosage while preserving the effect.

[0063] Table 4. Inhibition rate of tyrosinase activity by the combination of Dendrobium officinale leaf and Polygonatum sibiricum extract.

[0064]

[0065] As shown in Table 4, compared with the single use of Dendrobium officinale leaf extract and Polygonatum sibiricum extract, the combination of the two in different proportions improved the tyrosinase activity inhibition ability of Dendrobium officinale leaf extract at the same concentration to a certain extent, but the improvement rate was not significant. The tyrosinase activity inhibition ability of Polygonatum sibiricum extract was slightly reduced. Among the nine ratios, within the same concentration gradient, the tyrosinase activity inhibition rates increased by 43.47%, 49.7%, 40.85%, 34.48%, 31.65%, 33.84%, 69.57%, 54.45%, and 35.6%, respectively. The inhibition rate of Preparation Example 17 was the largest. Therefore, the ratio of Preparation Example 17 was used for subsequent experiments. Based on the results of each ratio, the combination of the two still has limitations compared with the single use, and further optimization of the compound and ratio is needed.

[0066] Table 5. Inhibition rate of tyrosinase activity by Dendrobium officinale leaf extract, Polygonatum sibiricum extract, β-arbutin and their compound extracts.

[0067] Preparation Example Inhibition rate / (%) Preparation Example 20 23.25±5.15 Preparation Example 21 35.35±4.95 Preparation Example 22 51.98±3.48 Preparation Example 23 37.19±12.07 Preparation Example 24 85.26 ± 2.36 ##** ]] Preparation Example 25 97.06 ± 4.56 ##** ]] Preparation Example 26 69.48 ± 2.28 ##* ]] Preparation Example 27 63.89±11.39 Preparation Example 28 91.96 ± 3.18 ##** ]]

[0068] Note: Compared with the use of Dendrobium officinale leaf-Polygonatum complex alone: # P<0.05, ## P<0.01;

[0069] Compared to using β-arbutin alone: * P<0.05, ** P<0.01

[0070] As shown in Table 5, at the same concentration, β-arbutin has a stronger inhibitory effect on tyrosinase activity than Polygonatum sibiricum and Dendrobium officinale leaves. When the three are combined in different proportions, they interact and combine organically, resulting in a significant enhancement of the tyrosinase inhibitory effect in Preparation Example 25. Compared with the same concentration, the inhibitory effects of Dendrobium officinale leaves, Polygonatum sibiricum, and β-arbutin on tyrosinase increased from 23.25%, 35.35%, and 51.98% to 97.06%, respectively, representing increases of approximately 300%, 175%, and 87%, respectively. The improvement effect is extremely significant.

[0071] Example 7

[0072] Zebrafish model for evaluating the inhibitory effect of the compound on melanin production

[0073] Fifteen zebrafish embryos, aged 6–8 hours and developing normally, were added to each well using 6-well plates. After the culture water was aspirated with plastic pipettes, 4.0 mL of a diluted sample solution of *Dendrobium officinale* leaf-*Polygonatum sibiricum*-β-arbutin was added to each well. The plates were incubated at a constant temperature of 28 ± 0.5 °C, with the culture medium changed every 12 hours. Three parallel control groups, a *Dendrobium officinale* leaf group, a *Polygonatum sibiricum* group, a β-arbutin group, and a *Dendrobium officinale* leaf-*Polygonatum sibiricum* group were also set up. The distribution of melanin granules in the embryos was observed and recorded under a microscope at 72 hours. The melanin content was statistically analyzed using Image-Pro Plus software. All experimental procedures followed the guidelines for laboratory animal care and use.

[0074] Table 6. Statistical table of relative melanin content in different groups of zebrafish.

[0075]

[0076]

[0077] like Figure 1As shown in the results, compared with the blank group, the melanin inhibition rates of the groups treated with Dendrobium officinale leaf, Polygonatum sibiricum, β-arbutin, Dendrobium officinale leaf-Polygonatum sibiricum, and Dendrobium officinale leaf-Polygonatum sibiricum-β-arbutin were 19.96%, 15.61%, 79.51%, 71.84%, and 98.70%, respectively. The compound can significantly inhibit melanin production compared with the blank group.

[0078] The above results show that the *Dendrobium officinale* leaf-*Polygonatum sibiricum*-β-arbutin complex of the present invention has a strong inhibitory ability on tyrosinase activity, which is significantly better than the individual use of the three extracts. Verification using a zebrafish model showed that it can effectively inhibit melanin production in zebrafish within 72 hours, significantly reducing melanin content. The melanin inhibition rate was 394%, 532%, 24.14%, and 37.39% higher than that of the *Dendrobium officinale* leaf group, the *Polygonatum sibiricum* group, the β-arbutin group, and the *Dendrobium officinale* leaf-*Polygonatum sibiricum* group, respectively. The *Dendrobium officinale* leaf-*Polygonatum sibiricum* complex can significantly improve the melanin inhibition rate in zebrafish, achieving the inhibitory effect of β-arbutin. The combination of these two with β-arbutin can achieve a melanin inhibition rate of 98.70%, exhibiting whitening and spot-removing effects.

[0079] The main features and advantages of the present invention have been described above. The above embodiments are some, but not all, of the embodiments of the present invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

Claims

1. The application of a complex containing Dendrobium officinale leaf extract in the preparation of a whitening and freckle-removing agent, characterized in that, The complex includes Dendrobium officinale leaf extract, Polygonatum sibiricum extract and β-arbutin, wherein the mass ratio of Dendrobium officinale leaf extract to Polygonatum sibiricum extract is 1:0.04-22.60, and the mass ratio of the total mass of Dendrobium officinale leaf extract and Polygonatum sibiricum extract to β-arbutin is 0.6-15.2:

1. The preparation method of Dendrobium officinale leaf extract is as follows: Dendrobium officinale leaves are dried in a forced-air dryer at 200℃, crushed by a pulverizer, soaked in a solvent (1%~80% ethanol aqueous solution) for 0.2~20 h, refluxed for extraction at 30~100℃ for 1~10 h, and extracted 1~15 times. After filtration, the filtrate is concentrated under reduced pressure at 20~70℃, and the concentrate is freeze-dried to obtain Dendrobium officinale leaf extract. The preparation method of Polygonatum extract is as follows: Polygonatum is dried at 200℃ by forced air drying, crushed by a pulverizer, soaked in a solvent (5%~75% ethanol aqueous solution) for 0.5~18 h, refluxed for extraction at 60~100℃ for 2~8 h, and extracted 1~10 times. After filtration, the filtrate is concentrated under reduced pressure at 20~80℃, and the concentrate is freeze-dried to obtain Polygonatum extract.

2. The application of the complex containing Dendrobium officinale leaf extract as described in claim 1 in the preparation of a whitening and freckle-removing preparation, characterized in that, The mass ratio of Dendrobium officinale leaf extract to Polygonatum sibiricum extract is 1:0.26-13.

80.

3. The application of the complex containing Dendrobium officinale leaf extract as described in claim 2 in the preparation of a whitening and freckle-removing preparation, characterized in that, The mass ratio of Dendrobium officinale leaf extract, Polygonatum sibiricum extract and β-arbutin was 25:78:10.

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