A synergistic composition of Terminalia chebula extract and vitamin C and derivatives thereof, and a preparation method and application thereof

Abstract

The present application belongs to the technical field of cosmetics, and particularly relates to a synergistic composition of Terminalia chebula extract and vitamin C and derivatives thereof, and a preparation method and application thereof. The present application provides a composition containing Terminalia chebula extract and vitamin C / vitamin C ethyl ether. By adjusting the proportion of Terminalia chebula extract in the composition, the obtained composition exhibits a significant synergistic effect in terms of antioxidant effect and inhibition of the activities of multiple biological enzymes such as hyaluronidase, tyrosinase, elastase and collagenase, compared with single components such as Terminalia chebula extract or vitamin C or vitamin C ethyl ether. Therefore, the composition can be used as a core raw material for preparing drugs, foods and cosmetics with the effects of delaying aging and whitening, greatly expanding the application range of Terminalia chebula extract and improving the application effect of Terminalia chebula extract, vitamin C and derivatives thereof.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, and specifically relates to a synergistic composition of Terminalia chebula extract and vitamin C and its derivatives, as well as its preparation method and application. Background Technology

[0002] As the largest organ in direct contact with the external environment, the skin is subjected to multiple external stimuli over a long period, such as sunlight, air pollutants (e.g., PM2.5), and chemical irritants. These factors can lead to excessive accumulation of reactive oxygen species (ROS) in the skin, disrupting the balance between the body's oxidation and antioxidant systems and triggering oxidative stress. This stress response directly damages skin cell DNA, lipids, and cell membranes, activates inflammatory pathways and promotes the release of inflammatory factors, while inhibiting the synthesis of type I and type III collagen and inducing increased expression of matrix metalloproteinases. Ultimately, this results in aging and damage symptoms such as deepened wrinkles, decreased elasticity, pigmentation, and sensitivity with redness. Therefore, developing skincare ingredients and compositions with both high antioxidant and anti-aging properties has become a core demand in the cosmetics industry.

[0003] Vitamin C and its derivatives are recognized as classic antioxidants in the skincare field. Their efficacy in neutralizing free radicals, inhibiting melanin production, and promoting collagen synthesis has been fully verified. However, the efficacy of vitamin C and its derivatives when used alone is singular and limited, making it difficult to simultaneously address issues such as inflammatory responses and collagen loss caused by oxidative stress.

[0004] Terminalia chebula, a classic medicinal herb in traditional Tibetan and Chinese medicine, is rich in hydrolyzable tannins (chebulic acid, chebulic acid, etc.), phenolic acids, and other active ingredients, and has a long history of medicinal use. Modern research has confirmed that Terminalia chebula has significant antioxidant, anti-inflammatory, and antibacterial activities. In recent years, the application potential of Terminalia chebula in the field of skincare has been gradually discovered, especially in regulating skin barrier function, inhibiting the expression of inflammatory factors, and delaying photoaging, showing good results.

[0005] Although the skincare benefits of Terminalia chebula extract have been verified, its application in cosmetics is still in its early stages. It is mostly used in simple combinations with other plant extracts, and there are no research reports on its use in combination with vitamin C and its derivatives to achieve synergistic effects. Summary of the Invention

[0006] To address the above technical problems, this invention proposes a synergistic composition of Terminalia chebula extract and vitamin C and its derivatives, as well as its preparation method and application.

[0007] During the experiment, the inventors of this invention unexpectedly discovered that when Terminalia chebula extract is used in combination with vitamin C and its derivatives in a specific ratio, it can exhibit significant synergistic effects in multiple aspects such as anti-oxidation, anti-aging, and whitening. The combination is superior to the use of a single ingredient in improving skin elasticity, reducing wrinkles, inhibiting the release of inflammatory mediators, and repairing photodamage. However, if the ratio of the two is not appropriate, not only will it fail to produce a synergistic effect, but it may even produce an antagonistic effect, and the final effect of the combination may not be as good as that of a single ingredient.

[0008] Based on this, the present invention provides a composition containing a specific mass ratio of Terminalia chebula extract and vitamin C and its derivatives, which has excellent antioxidant and whitening properties and is suitable for various skin care products, especially skin care products that have the effects of improving photoaging, uneven skin tone and damaged skin barrier.

[0009] The technical solution provided by this invention is as follows:

[0010] The first aspect of the present invention is to provide a synergistic composition of Terminalia chebula extract and vitamin C and its derivatives, the synergistic composition comprising: Terminalia chebula extract, vitamin C or a derivative of vitamin C, wherein the mass ratio of Terminalia chebula extract to vitamin C or vitamin C derivative is (1~5):(1~5);

[0011] The Terminalia chebula extract is prepared by a method comprising the following steps:

[0012] S1. After crushing the Terminalia chebula, the powder is sieved to obtain Terminalia chebula powder;

[0013] S2: Add the Terminalia chebula powder obtained in S1 to 8-12 times its mass of deionized water, soak at room temperature for 3-5 hours, and then extract ultrasonically at 40-50℃ and 500-700 W for 40-50 minutes. Filter to obtain the first filtrate and the residue. Add 6-9 times its mass of deionized water to the residue and extract ultrasonically again for the same time under the same conditions. Filter to obtain the second filtrate and the residue. Combine the first and second filtrates to obtain the Terminalia chebula extract.

[0014] S3 The Terminalia chebula extract obtained in S2 is filtered sequentially through a 0.5 μm ceramic microfiltration membrane and an organic ultrafiltration membrane with a molecular weight cutoff of 20 kDa. The microfiltration operation temperature is 40~50℃ and the operation pressure is 0.1~0.2 MPa. The ultrafiltration operation temperature is 30~40℃ and the operation pressure is 1.0~2.0 MPa. The resulting permeate is concentrated under reduced pressure and then freeze-dried to obtain the Terminalia chebula extract.

[0015] In the above-described synergistic composition, preferably, the vitamin C derivative is vitamin C ethyl ether.

[0016] As a further preferred embodiment, the Terminalia chebula extract is prepared by a method comprising the following steps:

[0017] S1. After crushing the Terminalia chebula, pass it through a 10-mesh sieve to obtain Terminalia chebula powder;

[0018] S2: The Terminalia chebula powder obtained in S1 is added to 10 times its mass of deionized water and soaked at room temperature for 4 hours. Then, it is ultrasonically extracted at 45℃ and 600 W for 45 minutes. After filtration, the first filtrate and the residue are obtained. Deionized water with a mass of 8 times the weight of the Terminalia chebula powder is added to the residue and ultrasonically extracted again at 45℃ and 600 W for 45 minutes. After filtration, the second filtrate and the residue are obtained. The first filtrate and the second filtrate are combined to obtain Terminalia chebula extract.

[0019] S3 The Terminalia chebula extract obtained in S2 was filtered sequentially through a 0.5 μm ceramic microfiltration membrane and an organic ultrafiltration membrane with a molecular weight cutoff of 20 kDa. The microfiltration operation temperature was 45℃ and the operating pressure was 0.1 MPa, and the ultrafiltration operation temperature was 35℃ and the operating pressure was 1.0 MPa. The resulting permeate was concentrated under reduced pressure and then freeze-dried to obtain the Terminalia chebula extract.

[0020] The second aspect of the present invention is to provide the application of the above-mentioned synergistic composition in the preparation of a product having any one of the following effects: antioxidant, inhibition of tyrosinase activity, inhibition of hyaluronidase activity, inhibition of collagenase activity, and inhibition of elastase activity. Preferably, the product is selected from cosmetics, food, pharmaceuticals, and health products. The synergistic composition is added to the product at a mass of 0.5% to 20%.

[0021] Preferably, in the application described, when the product has antioxidant effects, the synergistic composition consists of Terminalia chebula extract and vitamin C, with a mass ratio of Terminalia chebula extract to vitamin C of 5:1; or, the composition consists of Terminalia chebula extract and vitamin C ethyl ether, with a mass ratio of Terminalia chebula extract to vitamin C ethyl ether of 1:2.5.

[0022] Preferably, in the application described, when the product has the effect of inhibiting tyrosinase activity, the synergistic composition consists of Terminalia chebula extract and vitamin C, with a mass ratio of Terminalia chebula extract to vitamin C of 1:1; or, the composition consists of Terminalia chebula extract and vitamin C ethyl ether, with a mass ratio of Terminalia chebula extract to vitamin C ethyl ether of 5:1.

[0023] Preferably, in the application described, when the product has the effect of inhibiting hyaluronidase activity, the synergistic composition consists of Terminalia chebula extract and vitamin C, with a mass ratio of Terminalia chebula extract to vitamin C of 1:5; or, the synergistic composition consists of Terminalia chebula extract and vitamin C ethyl ether, with a mass ratio of Terminalia chebula extract to vitamin C ethyl ether of 2.5:1.

[0024] Preferably, in the application described, when the product has the effect of inhibiting collagenase activity, the synergistic composition consists of Terminalia chebula extract and vitamin C / vitamin C ethyl ether, wherein the mass ratio of Terminalia chebula extract to vitamin C / vitamin C ethyl ether is 1:5.

[0025] Preferably, in the application described, when the product has the effect of inhibiting elastase activity, the synergistic composition consists of Terminalia chebula extract and vitamin C / vitamin C ethyl ether, wherein the mass ratio of Terminalia chebula extract to vitamin C / vitamin C ethyl ether is 1:5.

[0026] The third aspect of the present invention is: providing a method for synergistically enhancing the antioxidant properties of vitamin C and its derivative vitamin C ethyl ether and its inhibitory effects on the activities of tyrosinase, hyaluronidase, collagenase, and elastase, specifically by adding Terminalia chebula extract to vitamin C and its derivative vitamin C ethyl ether, wherein the mass ratio of the added Terminalia chebula extract to the added vitamin C and its derivative vitamin C ethyl ether is 1~5:1~5.

[0027] Similarly, the present invention also provides a method for synergistically enhancing the antioxidant properties of Terminalia chebula extract and its inhibitory effects on tyrosinase, hyaluronidase, collagenase, and elastase activity, namely by adding vitamin C and its derivative vitamin C ethyl ether to Terminalia chebula extract, wherein the mass ratio of the added Terminalia chebula extract to the added vitamin C and its derivative vitamin C ethyl ether is 1~5:1~5.

[0028] The present invention has the following advantages and effects compared with the prior art:

[0029] (1) This invention provides a synergistic composition of Terminalia chebula extract and vitamin C and its derivatives. By adjusting the proportion of Terminalia chebula extract in the composition, the obtained composition shows a significant synergistic effect in terms of antioxidant effect and inhibition of the activities of various biological enzymes such as hyaluronidase, tyrosinase, elastase and collagenase compared with single components such as Terminalia chebula extract, vitamin C and vitamin C ethyl ether. Therefore, it can be used as a core raw material for preparing pharmaceuticals, food and cosmetics with anti-aging and whitening effects, which greatly expands the application range of Terminalia chebula extract and significantly improves the application effect of Terminalia chebula extract, vitamin C and its derivatives.

[0030] (2) The synergistic composition of Terminalia chebula extract and vitamin C and its derivatives provided by the present invention can be obtained by two ultrasonic-assisted extractions and membrane filtration. The preparation process is simple and the formula is simple, which is especially suitable for large-scale industrial production. Detailed Implementation

[0031] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0032] Unless otherwise specified, all reagents, pharmaceuticals, and instruments used in the following examples were obtained commercially.

[0033] The Terminalia chebula extracts used in all embodiments and comparative examples of this invention were obtained according to the following uniform preparation method:

[0034] S1 crushes Terminalia chebula into powder and passes it through a 10-mesh sieve to obtain Terminalia chebula powder raw material;

[0035] S2: The Terminalia chebula powder obtained in S1 is added to 10 times its mass of deionized water and soaked at room temperature for 4 hours to obtain a Terminalia chebula powder soaking solution. The Terminalia chebula powder soaking solution is subjected to ultrasonic extraction at a temperature of 45°C, a power of 600 W, and a time of 45 min. After filtration, a first filtrate and a filter residue are obtained. Deionized water with a mass of 8 times that of the Terminalia chebula powder is added to the filter residue, and ultrasonic extraction is performed again at a temperature of 45°C and a power of 600 W for 45 min. After filtration, a second filtrate and a filter residue are obtained. The first filtrate and the second filtrate are combined to obtain a Terminalia chebula extract.

[0036] S3 involves sequentially filtering the Terminalia chebula extract obtained in S2 through a 0.5 μm ceramic microfiltration membrane and an organic ultrafiltration membrane with a molecular weight cutoff of 20 kDa. After filtration, a permeate is obtained. The permeate is then concentrated under reduced pressure and freeze-dried to obtain the Terminalia chebula extract. The microfiltration membrane is operated at a temperature of 45°C and a pressure of 0.1 MPa, while the ultrafiltration membrane is operated at a temperature of 35°C and a pressure of 1.0 MPa.

[0037] Example 1

[0038] A synergistic skincare composition of Terminalia chebula extract and vitamin C, wherein the mass ratio of Terminalia chebula extract to vitamin C is 1:1, and the composition is obtained by mixing the components.

[0039] Example 2

[0040] A synergistic skincare composition of Terminalia chebula extract and vitamin C, wherein the mass ratio of Terminalia chebula extract to vitamin C is 1:2.5.

[0041] Example 3

[0042] A synergistic skincare composition of Terminalia chebula extract and vitamin C, wherein the mass ratio of Terminalia chebula extract to vitamin C is 2.5:1.

[0043] Example 4

[0044] A synergistic skincare composition of Terminalia chebula extract and vitamin C, wherein the mass ratio of Terminalia chebula extract to vitamin C is 1:5.

[0045] Example 5

[0046] A synergistic skincare composition of Terminalia chebula extract and vitamin C, wherein the mass ratio of Terminalia chebula extract to vitamin C is 5:1.

[0047] Example 6

[0048] A synergistic skincare composition of Terminalia chebula extract and vitamin C ethyl ether, wherein the mass ratio of Terminalia chebula extract to vitamin C ethyl ether is 1:1.

[0049] Example 7

[0050] A synergistic skincare composition of Terminalia chebula extract and vitamin C ethyl ether, wherein the mass ratio of Terminalia chebula extract to vitamin C is 1:2.5.

[0051] Example 8

[0052] A synergistic skincare composition of Terminalia chebula extract and vitamin C ethyl ether, wherein the mass ratio of Terminalia chebula extract to vitamin C ethyl ether is 2.5:1.

[0053] Example 9

[0054] A synergistic skincare composition of Terminalia chebula extract and vitamin C ethyl ether, wherein the mass ratio of Terminalia chebula extract to vitamin C ethyl ether is 1:5.

[0055] Example 10

[0056] A synergistic skincare composition of Terminalia chebula extract and vitamin C ethyl ether, wherein the mass ratio of Terminalia chebula extract to vitamin C ethyl ether is 5:1.

[0057] Comparative Example 1

[0058] This comparative example provides a product made from vitamin C.

[0059] Comparative Example 2

[0060] This comparative example provides a product made solely from Terminalia chebula extract, prepared using the same method as in Example 1.

[0061] Comparative Example 3

[0062] This comparative example provides a product made from vitamin C ethyl ether.

[0063] Example 1

[0064] This example demonstrates the antioxidant effects of the products prepared in the various embodiments and comparative examples of this invention.

[0065] This example demonstrates the effects of each sample on DPPH and ABTS. + Free radical scavenging effect, verification method and IC 50 The methods for obtaining the materials are completely consistent with those disclosed in patent CN119385891A, and will not be repeated in this invention.

[0066] In this invention, the Loewe equivalent linear additive model was used to evaluate the interaction type between Terminalia chebula extract and vitamin C and its derivatives. First, the IC50 values ​​of vitamin C (Comparative Example 1), Terminalia chebula extract (Comparative Example 2), and vitamin C ethyl ether (Comparative Example 3) were determined. 50 Based on this, the IC values ​​of each embodiment were measured. 50 Calculate the Combination Index (CI).

[0067] .

[0068] In the formula, d A d B These are the actual concentrations of Terminalia chebula extract and vitamin C or its derivatives when used in combination; IC50 of Terminalia chebula extract 50 , For vitamin C IC 50 CI<1, CI=1, and CI>1 were respectively determined as synergistic, additive, and antagonistic effects.

[0069] Furthermore, this invention employs the Bliss independent model and the King's method (Q-value method) to evaluate the interaction type of the combined use of Terminalia chebula extract and vitamin C: assuming that the effects of A and B are independent, the expected effect of the combined use is:

[0070] ;

[0071] In the formula, E A E BThese represent the actual effects (inhibition rates) of A and B individually at their respective doses when used in combination. If the actual combined effect... > If they are equal, they are synergistic; if they are equal, they are additive; if they are less than, they are antagonistic.

[0072] Based on the Bliss model, the synergy index Q is calculated for determination: Q = E comb / E bliss ;

[0073] A synergistic effect is defined as Q > 1.15, an additive effect is defined as 0.85 ≤ Q ≤ 1.15, and an antagonistic effect is defined as Q < 0.85.

[0074] Tables 1 and 2 show the DPPH radical scavenging rates of samples from Examples 1-5 and Comparative Examples 1-2, and Examples 6-10 and Comparative Examples 2-3, respectively; Tables 3 and 4 show the ABTS rates of samples from Examples 1-5 and Comparative Examples 1-2, and Examples 6-10 and Comparative Examples 2-3, respectively. + Free radical scavenging rate.

[0075] Table 1. DPPH radical scavenging rates of samples from Examples 1-5 and Comparative Examples 1-2 Example 1 1:1 19.38 0.342 1.768 Example 2 1:2.5 32.21 0.601 1.277 Example 3 2.5:1 12.07 0.201 2.682 Example 4 1:5 39.38 0.756 1.145 Example 5 5:1 5.63 0.090 5.037 Comparative Example 1 0:1 50.02 —— —— Comparative Example 2 1:0 65.45 —— ——

[0076] Table 1 shows the IC50 values ​​of the samples obtained from Comparative Example 1 and Comparative Example 2 for DPPH radical scavenging. 50 The concentrations were 50.02 μg / mL and 65.45 μg / mL, respectively, while the IC50 values ​​of the samples in each example were... 50 The concentrations ranged from 5.63 to 39.38 μg / mL, indicating that the combination of Terminalia chebula extract and vitamin C exhibited a definite synergistic effect. Furthermore, the CI values ​​of the compositions calculated using the Loewe model ranged from 0.090 to 0.756, all less than 1. The Bliss independent model and the King's method further validated this result; the actual combined effect of all formulations was greater than the expected effect, with Q values ​​ranging from 1.145 to 5.037. Except for Example 4, the Q values ​​of the other examples were all greater than the synergistic threshold of 1.15. Among them, Example 5 showed the most significant synergistic effect (CI = 0.090, Q = 5.037).

[0077] Table 2. DPPH radical scavenging rates of samples from Examples 6-10 and Comparative Examples 2-3

[0078] Table 2 shows that the CI values ​​of all compositions in each example were less than 1 (range 0.352~0.934), indicating that the combination of Terminalia chebula extract and Vc ethyl ether has a synergistic effect. Furthermore, the Q-value results using the King's method show that, except for Example 6 with a Q value of 0.934 (ranging from 0.85 to 1.15, indicating an additive effect), the Q values ​​of Examples 7 to 10 were all greater than 1.15, demonstrating a synergistic effect. Among them, Example 8 showed the strongest synergistic effect (CI=0.352, Q=1.844), while the equal-proportion mixture only exhibited an additive effect.

[0079] Table 3. ABTS of Samples from Examples 1-5 and Comparative Examples 1-2 + Free radical scavenging rate Example 1 1:1 35.66 0.444 1.515 Example 2 1:2.5 66.85 0.874 0.994 Example 3 2.5:1 29.31 0.347 1.836 Example 4 1:5 69.18 0.928 0.986 Example 5 5:1 16.20 0.186 3.105 Comparative Example 1 0:1 72.03 —— —— Comparative Example 2 1:0 90.69 —— ——

[0080] Table 3 shows that the CI values ​​of the samples in each embodiment ranged from 0.186 to 0.928, all less than 1, indicating that the formulations in each embodiment exhibited a synergistic effect. Furthermore, the Q values ​​of Examples 1, 3, and 5 were 1.515, 1.836, and 3.105 (all > 1.15), respectively, demonstrating a synergistic effect, while the Q values ​​of Examples 2 and 4 were 0.994 and 0.986 (within the range of 0.85 to 1.15), respectively, indicating an additive effect. Combining the two models, the synergistic effect of the composition in Example 5 was the most significant.

[0081] Table 4. ABTS of Samples from Examples 6-10 and Comparative Examples 2-3 + Free radical scavenging rate

[0082] Table 4 shows that the CI values ​​of all examples were less than 1 (range 0.506~0.884), indicating that each ratio in the examples had a synergistic effect. Furthermore, the Q values ​​of Examples 7-9 were all >1.15, indicating a synergistic effect, while the Q values ​​of Examples 6 and 10 were 0.970 and 1.144 respectively (within the range of 0.85~1.15), indicating an additive effect. Combining the two models, the Terminalia chebula extract and Vc ethyl ether showed a clear synergistic effect in scavenging ABTS at ratios of 1:2.5, 2.5:1, and 1:5. + The free radical effect was observed, with Example 7 showing the strongest synergistic effect, while the ratios in Examples 6 and 10 only exhibited additive effects.

[0083] Based on the results in Tables 1 to 4 above, it can be seen that the synergistic composition exhibits the best in vitro antioxidant effect when it is composed of Terminalia chebula extract and vitamin C in a mass ratio of 5:1, or when it is composed of Terminalia chebula extract and vitamin C ethyl ether in a mass ratio of 1:2.5.

[0084] Example 2

[0085] This example provides the whitening effect (evaluation of tyrosinase inhibition effect) of the samples prepared by various embodiments and comparative examples of the present invention.

[0086] The specific procedures for the tyrosinase inhibition rate experiment are as follows:

[0087] Preparation of sample solutions: The samples of each example and comparative example were prepared into test sample solutions of corresponding concentrations using DMSO;

[0088] Preparation of L-tyrosine substrate solution: L-tyrosine substrate was prepared into a 0.05% L-tyrosine solution using phosphate buffer solution at pH 6.8.

[0089] Preparation of tyrosinase solution: Tyrosinase was prepared into a 100 U / mL solution using phosphate buffer solution at pH 6.8.

[0090] Determination of tyrosinase inhibition rate: Accurately pipette the four reaction sample solutions (C1, C2, T1, and T2, excluding the tyrosinase solution) according to Table 5. Incubate at 37℃ for 10 min, then add the tyrosinase solution and react for another 10 min. Using C2 as the reference for C1, measure the absorbance A1 at 475 nm. Using T2 as the reference for T1, measure the absorbance A2. To compensate for systematic errors caused by parallel samples, 1 mL of DMSO was added to both C1 and C2. To eliminate absorbance errors at 475 nm, 1 mL of sample solution was also added to T2. Calculate the tyrosinase inhibition rate using the following formula and calculate the IC50 for each sample. 50 The results are shown in Tables 6 and 7. The formula for calculating the tyrosinase inhibition rate is as follows:

[0091] ;

[0092] Where A1 represents the change in absorbance of the reaction solution without the sample, and A2 represents the change in absorbance of the reaction solution after the sample is added.

[0093] Table 5. Reagent dosage for tyrosinase activity inhibition experiment

[0094] Table 6. Tyrosinase activity inhibition rate of samples from Examples 1-5 and Comparative Examples 1-2 Example 1 1:1 0.060 0.208 2.878 Example 2 1:2.5 0.131 0.629 1.284 Example 3 2.5:1 0.235 0.502 1.445 Example 4 1:5 0.054 0.299 2.165 Example 5 5:1 0.161 0.225 2.642 Comparative Example 1 0:1 0.152 —— —— Comparative Example 2 1:0 2.771 —— ——

[0095] Table 6 shows that the CI values ​​of all examples were less than 1 (range 0.208~0.629), and the Q values ​​of the King's Law were all greater than 1.15 (range 1.284~2.878), indicating that the ratios of all examples exhibited a synergistic inhibitory effect. Among them, Example 1 showed the most significant synergistic effect, followed by Example 5. That is, the combined use of Terminalia chebula extract and vitamin C at all ratios can significantly enhance tyrosinase inhibitory activity, with Examples 1 and 5 showing the most prominent synergistic effects.

[0096] Table 7. Tyrosinase activity inhibition rates of samples from Examples 6-10 and Comparative Examples 2-3

[0097] Table 7 shows the results: in the tyrosinase activity inhibition experiment, the IC50 of Comparative Example 3 was [missing information]. 50 The concentration was 0.390 mg / mL, while the IC50 of the compositions in each example was... 50 The CI values, ranging from 0.120 to 0.341, and the fact that all ratios in each example had CI values ​​less than 1, demonstrate a synergistic effect in the inhibition of tyrosinase activity. Furthermore, the Q values ​​of all examples were greater than 1.15, indicating synergy. Example 10 (Terminalia chebula extract:Vc ethyl ether = 5:1) showed the most significant synergistic effect, followed by Example 8.

[0098] The results from both models show that Terminalia chebula extract, vitamin C, or vitamin C ethyl ether exhibits a synergistic effect in inhibiting tyrosinase activity at all ratios. When Terminalia chebula extract and vitamin C are combined at a mass ratio of 1:1 or at a relatively high ratio (5:1, 2.5:1), the synergistic effect of the resulting composition is stronger, and the inhibitory effect on tyrosinase activity is most prominent.

[0099] Example 3

[0100] This example demonstrates the anti-aging effects (hyaluronidase inhibition effect evaluation) of the samples prepared in the various embodiments and comparative examples of this invention. Specific experimental procedures are as follows:

[0101] Take 50 μL of sample and mix it with 50 μL of hyaluronidase solution (15 μg / mL) and 12 μL of phosphate buffer (0.3 mol / L, pH 5.35). Add 50 μL of sodium hyaluronate solution (0.3 mg / mL) and shake to mix. Incubate at 37℃ for 45 min. Then add 100 μL of acidic albumin solution (1% BSA, dissolved in 0.1 mol / L acetate-sodium acetate buffer, pH 3.75), let stand at room temperature for 10 min, and measure the absorbance at 600 nm. At the same time, set up a sample background control (using buffer instead of enzyme solution), a blank control (using DMSO instead of sample), and a blank background control, and label them A, A0, B, and B0, respectively. Perform three parallel measurements and calculate the inhibition rate (%) as [1-(A-A0) / (B-B0)]×100%.

[0102] Table 8. Hyaluronidase activity inhibition rate of samples from Examples 1-5 and Comparative Examples 1-2 Example 1 1:1 0.490 0.392 1.775 Example 2 1:2.5 0.420 0.192 3.102 Example 3 2.5:1 0.561 0.641 1.280 Example 4 1:5 0.499 0.133 4.255 Example 5 5:1 0.617 0.824 1.107 Comparative Example 1 0:1 NA —— —— Comparative Example 2 1:0 0.625 —— ——

[0103] Table 8 shows that the extract of Terminalia chebula has an inhibitory effect on hyaluronidase, and its IC50 value is [missing information]. 50 The concentration was 0.625 mg / mL, while vitamin C did not show significant inhibitory activity (IC50). 50 (denoted as NA). When used together, the CI values ​​of each example are less than 1, and the Q values ​​of Examples 1 to 4 are all greater than 1.15, indicating that the combination of the two in the ratios of Examples 1 to 4 exhibits a synergistic inhibitory effect on hyaluronidase. The Q value of Example 5 is 1.107, which is between 0.85 and 1.15, indicating an additive effect.

[0104] Combining the two models, vitamin C alone does not inhibit hyaluronidase activity, but when used in combination, the addition of vitamin C can enhance the hyaluronidase inhibitory ability of Terminalia chebula extract, with the synergistic effect being most significant in Example 4.

[0105] Table 9. Hyaluronidase activity inhibition rate of samples from Examples 6-10 and Comparative Examples 2-3

[0106] Table 9 shows that Vc ethyl ether had no significant inhibitory activity within the tested concentration range (IC50). 50 The CI values ​​of all examples (denoted as NA) were less than 1 (range 0.147~0.235), and the Q values ​​of the King's method were all greater than 1.15 (range 2.626~3.891), indicating that each ratio showed a synergistic inhibitory effect. The synergistic effect of the composition of Example 8 (Terminalia chebula:Vc ethyl ether = 2.5:1) was the most significant, followed by Example 9.

[0107] Example of effect 4

[0108] This example demonstrates the anti-aging effects (evaluation of collagenase inhibition effect) of the samples prepared in the various embodiments and comparative examples of the present invention. The specific operating methods are as follows:

[0109] Collagenase solution (0.5 U / mL, enzyme activity defined using FALGPA as substrate) and substrate FALGPA solution (2 mM, dissolved in the same buffer) were prepared using Tris-HCl buffer (50 mM, pH 7.5, containing 5 mM CaCl2). Three groups were set up for the experiment: Sample group (S) – 50 μL of sample and 50 μL of enzyme solution were pre-incubated at 37℃ for 5 min, followed by the addition of 50 μL of substrate; Enzyme control group (C) – 50 μL of buffer was used instead of the sample, with the rest the same; Substrate blank group (B) – 50 μL of buffer was used instead of the enzyme, 50 μL of buffer was used instead of the sample, and then 50 μL of substrate was added. After mixing, the mixture was incubated at 37℃ for 90 min (the incubation time could be adjusted according to enzyme activity, ensuring the absorbance decrease ΔOD in the enzyme control group was between 0.2 and 0.5). Since the absorbance at 345 nm decreased linearly after FALGPA hydrolysis, no stop solution or chromogenic agent was needed; the absorbance of each well was directly measured at 345 nm. The assay was performed in triplicate. The inhibition rate (%) was calculated using the formula: [1 - (OD200%)] S -OD B ) / (OD C -OD B Calculated as 100% × 100%. If the sample has background absorption at 345 nm, a background control sample (using buffer instead of enzyme) needs to be added.

[0110] Table 10. Collagenase activity inhibition rate of samples from Examples 1-5 and Comparative Examples 1-2 Example 1 1:1 0.310 0.460 1.587 Example 2 1:2.5 0.750 0.636 1.286 Example 3 2.5:1 0.330 0.699 1.215 Example 4 1:5 0.364 0.180 3.277 Example 5 5:1 0.771 1.907 0.762 Comparative Example 1 0:1 NA —— —— Comparative Example 2 1:0 0.337 —— ——

[0111] Table 10 shows that Terminalia chebula extract has an inhibitory effect on collagenase activity (IC50). 50 The concentration of the active ingredient was 0.337 mg / mL, while vitamin C showed no significant inhibitory activity against the enzyme. When used in combination, the results of the Loewe equivalent line addition model and the King's method were consistent. The CI values ​​for Examples 1-4 were all less than 1, and the Q values ​​were all greater than 1.15, indicating a synergistic effect. Among these, Example 4 showed the most significant synergistic effect. In Example 5, the CI value was 1.907 (>1), and the Q value was 0.762 (<0.85), indicating an antagonistic effect when used together.

[0112] The above results indicate that an appropriate amount of vitamin C can act as a highly effective synergist to significantly enhance the inhibitory activity of Terminalia chebula extract on collagenase. However, if the proportion of Terminalia chebula extract is too high, it will reduce the efficacy of the combination of the two.

[0113] Table 11. Collagenase activity inhibition rate of samples from Examples 6-10 and Comparative Examples 2-3

[0114] Table 11 shows that Terminalia chebula extract has an inhibitory effect on collagenase activity (IC50). 50 The concentration of the active ingredient was 0.337 mg / mL, while vitamin C ethyl ether had no significant inhibitory activity against the enzyme. In addition, the CI values ​​of Examples 6 to 9 were all less than 1, and the Q values ​​of Examples 6 to 9 were all greater than 1.15, indicating that the two had a synergistic effect when combined in the ratios of Examples 6 to 9. The CI value of Example 10 was 1.056 > 1, and the Q value was 0.973 (ranging from 0.85 to 1.15), showing an antagonistic or additive effect.

[0115] Combining the two models, Terminalia chebula extract and Vc ethyl ether exhibit synergistic inhibitory effects on hyaluronidase activity at ratios of 1:1, 1:2.5, 2.5:1, and 1:5. The synergistic effect is strongest under the ratio conditions of Example 9, while a high ratio of Terminalia chebula extract (5:1) shows antagonistic or additive effects. This suggests that appropriately increasing the ratio of Vc ethyl ether or keeping the ratio of Terminalia chebula extract below 2.5:1 is beneficial to improving the inhibitory effect on hyaluronidase activity.

[0116] Example 5

[0117] This example demonstrates the anti-aging effects (evaluation of elastase inhibition) of the samples prepared in the various embodiments and comparative examples of this invention. Specific experimental procedures are as follows:

[0118] Prepare 1.015 mmol / L AAAPAN and 6 U / mL elastase solutions using 50 mmol / L Tris-HCl buffer (pH 8.0). Two groups were set up: Sample Group (B) – 15 μL of sample solutions of different concentrations were added to 25 μL of AAAPAN solution, mixed, and incubated at 25°C for 10 min. Then, 25 μL of elastase solution was added to initiate the reaction, and incubation continued at 25°C for another 10 min. Control Group (A) – 15 μL of buffer solution was used instead of the sample solution, with the other procedures identical. Absorbance was measured at 410 nm immediately after the reaction. Each group was measured in triplicate, and the average value was taken. The inhibition rate was calculated using the formula: Inhibition rate (%) = (1-B / A) × 100%, where A is the absorbance of the control group and B is the absorbance of the sample group. If the sample solution itself absorbs at 410 nm, a background control (using buffer solution instead of enzyme solution) should be added to subtract the absorption.

[0119] Table 12. Elastase activity inhibition rate of samples from Examples 1-5 and Comparative Examples 1-2 Example 1 1:1 0.892 0.549 1.410 Example 2 1:2.5 0.444 0.156 3.699 Example 3 2.5:1 1.414 1.244 0.902 Example 4 1:5 0.409 0.084 6.463 Example 5 5:1 1.205 1.236 0.904 Comparative Example 1 0:1 NA —— —— Comparative Example 2 1:0 0.812 —— ——

[0120] Table 13 Elastase activity inhibition rate of samples from Examples 6-10 and Comparative Examples 2-3

[0121] The results in Tables 12-13 show that Terminalia chebula extract inhibits the IC50 of elastase. 50 The concentration was 0.812 mg / mL, and vitamin C and its derivatives had no significant inhibitory activity against this enzyme (IC50). 50 (denoted as NA). When used in combination, the results of the Loewe equivalent line addition model and the King's method for elastase inhibition were basically consistent. Specifically, when Terminalia chebula extract was combined with vitamin C, it showed a significant synergistic effect at the ratios in Examples 1, 2, and 4, while antagonistic effects were observed at the ratios in Examples 3 and 5. When Terminalia chebula extract was combined with vitamin C derivatives, the ratios in Examples 6-10 all showed a synergistic effect. The optimal synergistic ratios for elastase inhibition were the ratios in Examples 4 and 9, respectively, for the composition of Terminalia chebula extract with vitamin C and vitamin C ethyl ether. A high proportion of Terminalia chebula extract was detrimental to the synergistic effect of elastase inhibition.

[0122] The above are merely preferred embodiments of the present invention and do not limit the scope of the patent. All equivalent changes and modifications made within the scope of the present invention should still fall within the scope of the present invention.

Claims

1. A synergistic composition of Terminalia chebula extract and vitamin C and its derivatives, characterized in that, include: Terminalia chebula extract, vitamin C or a derivative of vitamin C, wherein the mass ratio of the Terminalia chebula extract to vitamin C or a derivative of vitamin C is (1~5):(1~5). The Terminalia chebula extract is prepared by a method comprising the following steps: S1. After crushing the Terminalia chebula, the powder is sieved to obtain Terminalia chebula powder; S2: Add the Terminalia chebula powder obtained in S1 to 8-12 times its mass of deionized water, soak at room temperature for 3-5 hours, and then extract ultrasonically at 40-50℃ and 500-700 W for 40-50 minutes. Filter to obtain the first filtrate and the residue. Add 6-9 times its mass of deionized water to the residue and extract ultrasonically again for the same time under the same conditions. Filter to obtain the second filtrate and the residue. Combine the first and second filtrates to obtain the Terminalia chebula extract. S3 The Terminalia chebula extract obtained in S2 is filtered sequentially through a 0.5 μm ceramic microfiltration membrane and an organic ultrafiltration membrane with a molecular weight cutoff of 20 kDa. The microfiltration operation temperature is 40~50℃ and the operation pressure is 0.1~0.2 MPa. The ultrafiltration operation temperature is 30~40℃ and the operation pressure is 1.0~2.0 MPa. The resulting permeate is concentrated under reduced pressure and then freeze-dried to obtain the Terminalia chebula extract.

2. The synergistic composition as claimed in claim 1, characterized in that, The vitamin C derivative is vitamin C ethyl ether.

3. The use of the synergistic composition as described in claim 1 in the preparation of a product having any one of the following effects: antioxidant, inhibition of tyrosinase activity, inhibition of hyaluronidase activity, inhibition of collagenase activity, and inhibition of elastase activity; The product is selected from any one of cosmetics, food, medicine, and health products; the synergistic combination is added to the product at a mass of 0.5% to 20%.

4. The application as described in claim 3, characterized in that, When the product has antioxidant effects, the synergistic composition consists of Terminalia chebula extract and vitamin C, with a mass ratio of Terminalia chebula extract to vitamin C of 5:1; or, the synergistic composition consists of Terminalia chebula extract and vitamin C ethyl ether, with a mass ratio of Terminalia chebula extract to vitamin C ethyl ether of 1:2.

5.

5. The application as described in claim 3, characterized in that, When the product has the effect of inhibiting tyrosinase activity, the synergistic composition consists of Terminalia chebula extract and vitamin C, with a mass ratio of Terminalia chebula extract to vitamin C of 1:1; or, the synergistic composition consists of Terminalia chebula extract and vitamin C ethyl ether, with a mass ratio of Terminalia chebula extract to vitamin C ethyl ether of 5:

1.

6. The application as described in claim 3, characterized in that, When the product has the effect of inhibiting hyaluronidase activity, the synergistic composition consists of Terminalia chebula extract and vitamin C, with a mass ratio of Terminalia chebula extract to vitamin C of 1:5; or, the synergistic composition consists of Terminalia chebula extract and vitamin C ethyl ether, with a mass ratio of Terminalia chebula extract to vitamin C ethyl ether of 2.5:

1.

7. The application as described in claim 3, characterized in that, When the product has the effect of inhibiting collagenase activity, the synergistic composition consists of Terminalia chebula extract and vitamin C / vitamin C ethyl ether, wherein the mass ratio of Terminalia chebula extract to vitamin C / vitamin C ethyl ether is 1:

5.

8. The application as described in claim 3, characterized in that, When the product has the effect of inhibiting elastase activity, the synergistic composition consists of Terminalia chebula extract and vitamin C / vitamin C ethyl ether, wherein the mass ratio of Terminalia chebula extract to vitamin C / vitamin C ethyl ether is 1:5.

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