Composition prepared by taking acacia and licorice as main materials, preparation method and application

Abstract

The present application belongs to the technical field of traditional Chinese medicine composition preparation, and particularly relates to a composition prepared by using chebula and licorice as main materials, and also relates to a preparation method of the composition and application of the composition in bacteriostatic anti-inflammatory, antioxidant, whitening, anti-glycation and anti-itching products. The composition of the present application comprises chebula and licorice as main materials; the composition is obtained by crushing, enzymolysis, water extraction, concentration, alcohol extraction and purification of chebula and licorice. Different combination schemes of chebula and licorice are provided in the present application based on different effects, so that the effects of the combination of chebula and licorice in different proportions are more significant; and it is also verified that the combination of chebula and licorice in different proportions has certain differences when they are applied in different products. The present application provides preparation methods of products with different effects suitable for the above-mentioned reasonable combination of chebula and licorice.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine composition preparation technology, specifically relating to a composition prepared mainly from Terminalia chebula and Glycyrrhiza uralensis, a method for preparing the above composition, and the application of the composition in antibacterial, anti-inflammatory, antioxidant, whitening, anti-glycation, and antipruritic products. Background Technology

[0002] Terminalia chebula is a traditional Tibetan medicine containing active ingredients such as gallic acid, chebulic acid, and chebulic acid, as well as tannins and phenolic acids, triterpenoids such as ajanglamin, ajanglaminic acid, and fenestrated flavonoids such as rutin and quercetin. These active ingredients endow Terminalia chebula with anti-inflammatory, analgesic, antimicrobial, antioxidant, hypoglycemic, anticholinesterase activity, and anti-gastric ulcer effects.

[0003] Licorice, with its sweet and neutral properties, contains glycyrrhizic acid and glycyrrhizin, as well as various flavonoids such as glycyrrhizin, isoglycyrrhizin, and polysaccharides. Licorice has the effects of tonifying the spleen and replenishing qi, clearing heat and detoxifying, relieving spasms and pain, and harmonizing other medicines. It is mainly used for spleen and stomach weakness, fatigue, palpitations, shortness of breath, cough with excessive phlegm, abdominal and limb spasms and pain, carbuncles and boils, and mitigating the toxicity of other drugs. Modern pharmacological studies have shown that licorice has various effects, including anti-inflammatory, antiviral, antioxidant, immunomodulatory, anti-atherosclerotic, and anti-tumor properties.

[0004] Regarding the efficacy of combining Terminalia chebula and licorice, there is currently little literature reporting or disclosure on this topic. While both Terminalia chebula and licorice possess certain anti-inflammatory, whitening, or antibacterial effects, their individual effects are limited. Whether combining them will produce a synergistic effect is unknown and requires further investigation; or, in other words, whether combining them will generate new effects is also unknown. Furthermore, given that both Terminalia chebula and licorice contain multiple active ingredients with varying effects, concentrating the components possessing specific effects would yield more significant results. Research in this area is also scarce. Summary of the Invention

[0005] To address the aforementioned technical issues, this application conducted a detailed study on the combined extraction and use of Terminalia chebula and licorice. The results showed that the efficacy and effects varied depending on the combination ratio of the two. Therefore, the greatest contribution of this application is providing a formulation suitable for a specific efficacy. For example, the ratio of Terminalia chebula to licorice in whitening products differs significantly from that in antioxidant products.

[0006] In this invention, the proportions of Terminalia chebula and licorice are as follows, depending on the different uses of the composition:

[0007] A composition prepared mainly from Terminalia chebula and Glycyrrhiza uralensis, wherein the main ingredients of the composition include Terminalia chebula and Glycyrrhiza uralensis; the preparation method is to pulverize Terminalia chebula and Glycyrrhiza uralensis, enzymatically hydrolyze, extract with water, concentrate, decolorize, and dilute with polyol to obtain the composition product.

[0008] Although both use Terminalia chebula and licorice as the main ingredients and employ the same method to prepare the composition, the efficacy of the resulting product varies significantly depending on the ratio of Terminalia chebula to licorice. For example:

[0009] When the composition is used to prepare an antibacterial and anti-inflammatory product, the mass ratio of Terminalia chebula to licorice is 2.5:1.

[0010] When the composition is used to prepare an antioxidant product, the mass ratio of Terminalia chebula to licorice is 5:1.

[0011] When the composition is used to prepare skin whitening products, the mass ratio of Terminalia chebula and licorice is 1:5.

[0012] When the composition is used to prepare anti-glycation products, the mass ratio of Terminalia chebula to licorice is 7:1.

[0013] When the composition is used to prepare antipruritic products, the mass ratio of Terminalia chebula to licorice is 10:1. This application also emphasizes the protection that different ratios of Terminalia chebula and licorice have different efficacies.

[0014] The method for preparing the above composition includes the following steps:

[0015] S1. After pulverizing Terminalia chebula and Glycyrrhiza uralensis, the powder is sieved to obtain powdered raw materials; wherein the mass ratio of Terminalia chebula to Glycyrrhiza uralensis is 2.5:1; or 5:1; or 1:5; or 7:1; or 10:1.

[0016] S2 involves adding deionized water to the powdered raw material in S1, adjusting the pH value to 4.0–6.0, and soaking it at 35–65°C for 30–90 minutes.

[0017] S3 is prepared by adding cellulase to the soaking solution of S2 and then enzymatically hydrolyzing it.

[0018] S4 takes the enzymatic hydrolysate from S3 and directly boils and extracts it, then filters it to obtain the first filtrate; the residue is then added to deionized water, boiled and extracted, then filtered to obtain the second filtrate; the first filtrate and the second filtrate are combined and filtered through a 0.22μm to 10μm filter membrane to obtain the extract;

[0019] S5 concentrates the extract from S4 to obtain a concentrated solution;

[0020] S6 Add activated carbon to the concentrate, stir, remove the activated carbon, and dry the resulting supernatant to obtain the extract.

[0021] S7 involves adding deionized water and polyol to the extract from S6, stirring to dissolve the extract, letting it stand overnight, taking the supernatant, drying it, and obtaining the composition.

[0022] The amount of deionized water and polyol added is 1 to 2 times the weight of the powder raw material;

[0023] The polyol is selected from any one of 1,2-propanediol, 1,3-propanediol, butanediol, dipropylene glycol, and glycerol.

[0024] In the above method, the preferred parameters in each step are:

[0025] S1 passes through a 10-20 mesh sieve;

[0026] S2 was adjusted to pH 4.0–6.0 using a 10 wt% hydrochloric acid solution;

[0027] S3. Add cellulase to the soaking solution of S2. The amount of cellulase added is 0.2-1.0% of the mass of the medicinal powder raw material. The enzymatic hydrolysis reaction is carried out in a water bath at 35-65℃ for 30-120 minutes.

[0028] In S4, the enzymatic extract of S3 is boiled and extracted for 1-3 hours, filtered, and the filtrate is reserved. The residue is then added to 5-20 times the weight of the herbal powder raw material in deionized water, boiled and extracted for 1-3 hours, filtered, and the filtrates are combined and filtered through a 0.22μm-10μm filter membrane to obtain the filtrate.

[0029] In S5, the extract is concentrated at 60–80°C until a concentrate of 0.2–0.5 g crude drug / mL is obtained;

[0030] In S6, activated carbon is added at a ratio of 1 to 50 mg of activated carbon per liter of concentrated liquid, and the mixture is stirred at 40 to 80°C for 2 to 5 hours. Then, the activated carbon is removed, and the supernatant is dried to obtain the extract.

[0031] This invention also provides the application of the above-described composition or the composition prepared by the above-described method in the preparation of products with antibacterial, anti-inflammatory, antioxidant, whitening, antipruritic, and anti-glycation effects. These applications are also the focus of this invention. Specific application effects are shown in the effect verification section of the examples.

[0032] For example, the mass ratio of Terminalia chebula to licorice is 2.5:1, which can be used in antibacterial and anti-inflammatory products. These products can be personal care products or oral hygiene products, such as facial cleanser, face cream, sunscreen, isolation cream, toothpaste, mouth spray, mouthwash, etc. Of course, these products are just examples.

[0033] When the mass ratio of Terminalia chebula to licorice is 5:1, it can be used in food or skin care products.

[0034] When the mass ratio of Terminalia chebula to licorice is 10:1, it can be used in antipruritic ointments or antipruritic shampoo and conditioner, such as antipruritic shampoos and conditioner solutions.

[0035] When the mass ratio of Terminalia chebula and licorice is 1:5, the composition can be used in whitening products, such as skin care products like face creams and masks.

[0036] When the mass ratio of Terminalia chebula to licorice is 7:1, the composition can be used in anti-glycation products.

[0037] The beneficial effects of this invention are as follows:

[0038] (1) Based on different effects, this invention provides different combination schemes of Terminalia chebula and Glycyrrhiza uralensis, so that their effects are more significant after being combined in different proportions; and it also verifies that after different proportions of Terminalia chebula and Glycyrrhiza uralensis are combined, they have certain differences when applied in different products; a composition with a certain effect corresponds to a specific proportion of Terminalia chebula and Glycyrrhiza uralensis; in this application, different effects were verified through effect examples 1-6.

[0039] (2) Based on the reasonable combination of Terminalia chebula and licorice, this application also provides a method for preparing products with different effects suitable for these formulas. In this application, the active ingredients in Terminalia chebula and licorice are fully dissolved by enzymatic hydrolysis, which helps to better exert the role of the active ingredients in both.

[0040] (3) Compared with complex formulas composed of dozens of medicinal materials, the formula in this application is simple, with only two medicinal materials, making the preparation process more convenient and simple. Attached Figure Description

[0041] Figure 1 The results of the inhibition zone determination for Staphylococcus aureus in Example 1 are shown.

[0042] Figure 2 The results of the inhibition zone determination for Trichophyton mentagrophytes in Example 1;

[0043] Figure 3 The results of the inhibition zone determination for Candida albicans in Example 1 are shown.

[0044] Figure 4 The results of the inhibition zone determination for Streptococcus mutans in Example 1 are shown.

[0045] Figure 5 The results of the inhibition zone assay for Porphyromonas gingivalis in Example 1 are shown.

[0046] Figure 6 The results of the DPPH removal rate test in Example 2;

[0047] Figure 7 The results of the ABTS clearance rate test in Example 2;

[0048] Figure 8 The anti-inflammatory test results are from Example 3;

[0049] Figure 9 The results are from the experiment on the tyrosinase inhibition rate in Example 4. Detailed Implementation

[0050] To further illustrate the solution and effects of the present invention, the following specific embodiments are provided.

[0051] I. Examples of Preparation Methods

[0052] Example 1

[0053] The main ingredients, Terminalia chebula and Glycyrrhiza uralensis, are in a mass ratio of 1:1.

[0054] The preparation method of this composition is as follows:

[0055] S1. After separately pulverizing Terminalia chebula and Glycyrrhiza uralensis, mix them in a 1:1 mass ratio and pass them through a 20-mesh sieve to obtain powder raw materials;

[0056] S2. Add the powdered raw materials in S2 to deionized water at a mass-to-volume ratio of 1:20, adjust the pH value to 4.0 with 10% hydrochloric acid solution, and soak in a 50℃ water bath for 60 minutes.

[0057] S3. Add cellulase to the soaking solution of S2. The amount of cellulase added is 0.6% of the mass of the medicinal powder raw material; and carry out the enzymatic hydrolysis reaction in a water bath at 50°C for 90 minutes.

[0058] S4. Boil the enzymatic extract from S3 for 1.5 hours, filter, and set aside the filtrate. Add 20 times the weight of the herbal powder in deionized water to the residue, boil for 1.5 hours, filter, combine the filtrates, and filter through a 5μm filter membrane to obtain the filtrate.

[0059] S5. Concentrate the filtrate from S4 under reduced pressure at 70℃ until the concentration of the drug solution is 0.5g crude drug / mL;

[0060] S6. Add 20 mg / mL of activated carbon to the concentrated solution of S5 according to the volume of the concentrated solution. Stir at 60°C for 3 hours, remove the activated carbon, and obtain the supernatant. After drying, obtain the extract.

[0061] S7. Add 1.5 times the amount of deionized water and 1.5 times the amount of 1,2-propanediol to the extract obtained in S6, stir to dissolve, let stand overnight, and take the supernatant to obtain the traditional Chinese medicine composition of the present invention.

[0062] Example 2

[0063] The main raw materials, Terminalia chebula and Glycyrrhiza uralensis, are in a mass ratio of 10:1, and can be used in the preparation of antipruritic products.

[0064] The preparation method is as follows:

[0065] S1. After separately pulverizing Terminalia chebula and Glycyrrhiza uralensis, mix them in a mass ratio of 10:1, pass them through a 10-mesh sieve, and obtain powder raw materials;

[0066] S2. Add the powdered raw materials in S2 to deionized water at a mass-volume ratio of 1:10, adjust the pH value to 5.0 with 10% hydrochloric acid solution, and soak in a 65℃ water bath for 30 minutes.

[0067] S3. Add cellulase to the soaking solution of S2. The amount of cellulase added is 0.2% of the mass of the medicinal powder raw material; and carry out the enzymatic hydrolysis reaction in a water bath at 65°C for 30 minutes.

[0068] S4. Boil the S3 enzymatic hydrolysis extract for 1 hour, filter, and set aside the filtrate. Add 10 times the weight of the herbal powder raw material in deionized water to the filter residue, boil for 1 hour, filter, combine the filtrates, and filter through a 10μm filter membrane to obtain the filtrate.

[0069] S5. Concentrate the filtrate from S4 under reduced pressure at 60℃ until the concentration of the drug solution is 0.2g crude drug / mL;

[0070] S6. Add 50 mg / mL of activated carbon to the concentrated solution of S5 according to the volume of the concentrated solution. Stir at 80°C for 2 hours, remove the activated carbon, and obtain the supernatant. After drying, obtain the extract.

[0071] S7. Add twice the amount of deionized water and twice the amount of 1,3-propanediol of the medicinal materials to the extract obtained in S6, stir to dissolve, let stand overnight, and take the supernatant to obtain the traditional Chinese medicine composition of the present invention.

[0072] Example 3

[0073] The main raw materials, Terminalia chebula and Glycyrrhiza uralensis, have a mass ratio of 2.5:1, and can be used in the preparation of antibacterial and anti-inflammatory products.

[0074] The preparation method of this composition is as follows:

[0075] S1. After separately pulverizing Terminalia chebula and Glycyrrhiza uralensis, mix them in a mass ratio of 2.5:1 and pass them through a 20-mesh sieve to obtain powder raw materials;

[0076] S2. Add the powdered raw materials from S1 to deionized water at a mass-volume ratio of 1:5, adjust the pH value to 4.0 with 10% hydrochloric acid solution, and soak in a 45℃ water bath for 60 minutes.

[0077] S3. Add cellulase to the soaking solution of S2. The amount of cellulase added is 1.0% of the mass of the medicinal powder raw material; and carry out the enzymatic hydrolysis reaction in a water bath at 45°C for 90 minutes.

[0078] S4. Boil the enzymatic hydrolysis extract from S3 for 2 hours, filter, and set aside the filtrate. Add 5 times the weight of the herbal powder raw material in deionized water to the residue, boil for 2 hours, filter, combine the filtrates, and filter through a 1μm filter membrane to obtain the filtrate.

[0079] S5. Concentrate the filtrate from S4 under reduced pressure at 70℃ until the concentration of the drug solution is 0.3g crude drug / mL;

[0080] S6. Add 20 mg / mL of activated carbon to the concentrated solution of S5 according to the volume of the concentrated solution. Stir at 60°C for 3.5 h, remove the activated carbon, and obtain the supernatant. After drying, obtain the extract.

[0081] S7. Add one times the mass of deionized water and one times the mass of butylene glycol to the extract obtained in S6, stir to dissolve, let stand overnight, and take the supernatant to obtain the traditional Chinese medicine composition of the present invention.

[0082] Example 4

[0083] The main ingredients, Terminalia chebula and Glycyrrhiza uralensis, are in a mass ratio of 1:10.

[0084] The preparation method of this composition is as follows:

[0085] S1. After separately pulverizing Terminalia chebula and Glycyrrhiza uralensis, mix them in a mass ratio of 1:10 and pass them through a 20-mesh sieve to obtain powder raw materials;

[0086] S2. Add the powdered raw materials from S1 to deionized water at a mass-volume ratio of 1:25, adjust the pH value to 5.5 with 10% hydrochloric acid solution, and soak in a 55℃ water bath for 30 minutes.

[0087] S3. Add cellulase to the S2 soaking solution. The amount of cellulase added is 0.6% of the mass of the medicinal powder raw material. Perform enzymatic hydrolysis reaction in a 40℃ water bath for 110 minutes.

[0088] S4. Boil the S3 enzymatic hydrolysis extract for 1.5 hours, filter, and set aside the filtrate. Add 15 times the weight of the herbal powder raw material in deionized water to the filter residue, boil for 1 hour, filter, combine the filtrates, and filter through a 6μm filter membrane to obtain the filtrate.

[0089] S5. Concentrate the filtrate from S4 under reduced pressure at 65℃ until the concentration of the drug solution is 0.25g crude drug / mL;

[0090] S6. Add 35 mg / mL of activated carbon to the concentrated solution of S5 according to the volume of the concentrated solution. Stir at 45°C for 4 hours, remove the activated carbon, and obtain the supernatant. After drying, obtain the extract.

[0091] S7. Add 1.5 times the mass of the medicinal material to deionized water and 1.5 times the mass of the medicinal material to dipropylene glycol, stir to dissolve, let stand overnight, and take the supernatant to obtain the traditional Chinese medicine composition of the present invention.

[0092] Example 5

[0093] The main raw materials, Terminalia chebula and Glycyrrhiza uralensis, are in a mass ratio of 1:5, and can be used in the preparation of skin whitening products.

[0094] The preparation method of this composition is as follows:

[0095] S1. After separately pulverizing Terminalia chebula and Glycyrrhiza uralensis, mix them in a mass ratio of 1:5, pass them through a 20-mesh sieve, and obtain powder raw materials;

[0096] S2. Add the powdered raw materials from S1 to deionized water at a mass-to-volume ratio of 1:20, adjust the pH value to 4.5 with 10% hydrochloric acid solution, and soak in a 60℃ water bath for 70 minutes.

[0097] S3. Add cellulase to the soaking solution of S2. The amount of cellulase added is 0.3% of the mass of the medicinal powder raw material; and carry out the enzymatic hydrolysis reaction in a water bath at 55°C for 70 minutes.

[0098] S4. Boil the enzymatic extract from S3 for 2.5 hours, filter, and set aside the filtrate. Add 20 times the weight of the herbal powder in deionized water to the residue, boil for 2.5 hours, filter, combine the filtrates, and filter through a 0.45μm filter membrane to obtain the filtrate.

[0099] S5. Concentrate the filtrate from S4 under reduced pressure at 75°C until the concentration of the drug solution is 0.45 g crude drug / mL;

[0100] S6. Add 25 mg / mL of activated carbon to the concentrated solution of S5 according to the volume of the concentrated solution. Stir at 75°C for 5 hours, remove the activated carbon, and obtain the supernatant. After drying, obtain the extract.

[0101] S7. Add one times the mass of the medicinal material to deionized water and one times the mass of the medicinal material to the extract obtained in S6, stir to dissolve, let stand overnight, and take the supernatant to obtain the traditional Chinese medicine composition of the present invention.

[0102] Example 6

[0103] The main raw materials, Terminalia chebula and Glycyrrhiza uralensis, are in a mass ratio of 5:1, and can be used in the preparation of antioxidant products.

[0104] The preparation method of this composition is as follows:

[0105] S1. After separately pulverizing Terminalia chebula and Glycyrrhiza uralensis, mix them in a mass ratio of 5:1 and pass them through a 20-mesh sieve to obtain powder raw materials;

[0106] S2. Add the powdered raw materials from S1 to deionized water at a mass-to-volume ratio of 1:20, adjust the pH value to 4.0 with 10% hydrochloric acid solution, and soak in a 50°C water bath for 60 minutes.

[0107] S3. Add cellulase to the soaking solution of S2. The amount of cellulase added is 0.6% of the mass of the medicinal powder raw material; and carry out the enzymatic hydrolysis reaction in a water bath at 50°C for 90 minutes.

[0108] S4. Boil the enzymatic extract from S3 for 1.5 hours, filter, and set aside the filtrate. Add 20 times the weight of the herbal powder in deionized water to the residue, boil for 1.5 hours, filter, combine the filtrates, and filter through a 5μm filter membrane to obtain the filtrate.

[0109] S5. Concentrate the filtrate from S4 under reduced pressure at 70℃ until the concentration of the drug solution is 0.5g crude drug / mL;

[0110] S6. Add 20 mg / mL of activated carbon to the concentrated solution of S5 according to the volume of the concentrated solution. Stir at 60°C for 3 hours, remove the activated carbon, and obtain the supernatant. After drying, obtain the extract.

[0111] S7. Add 1.5 times the amount of deionized water and 1.5 times the amount of 1,2-propanediol to the extract obtained in S6, stir to dissolve, let stand overnight, and take the supernatant to obtain the traditional Chinese medicine composition of the present invention.

[0112] Example 7

[0113] The main ingredients, Terminalia chebula and Glycyrrhiza uralensis, are in a mass ratio of 7:1 and can be used in anti-glycation products.

[0114] The preparation method is as follows:

[0115] S1. After separately pulverizing Terminalia chebula and Glycyrrhiza uralensis, mix them in a mass ratio of 7:1 and pass them through a 10-mesh sieve to obtain powder raw materials;

[0116] S2. Add the powdered raw materials from S1 to deionized water at a mass-to-volume ratio of 1:10, adjust the pH value to 5.0 with 10% hydrochloric acid solution, and soak in a 65℃ water bath for 30 minutes.

[0117] S3. Add cellulase to the soaking solution of S2. The amount of cellulase added is 0.2% of the mass of the medicinal powder raw material; and carry out the enzymatic hydrolysis reaction in a water bath at 65°C for 30 minutes.

[0118] S4. Boil the S3 enzymatic hydrolysis extract for 1 hour, filter, and set aside the filtrate. Add 10 times the weight of the herbal powder raw material in deionized water to the filter residue, boil for 1 hour, filter, combine the filtrates, and filter through a 10μm filter membrane to obtain the filtrate.

[0119] S5. Concentrate the filtrate from S4 under reduced pressure at 60℃ until the concentration of the drug solution is 0.2g crude drug / mL;

[0120] S6. Add 50 mg / mL of activated carbon to the concentrated solution of S5 according to the volume of the concentrated solution. Stir at 80°C for 2 hours, remove the activated carbon, and obtain the supernatant. After drying, obtain the extract.

[0121] S7. Add twice the amount of deionized water and twice the amount of 1,3-propanediol of the medicinal materials to the extract obtained in S6, stir to dissolve, let stand overnight, and take the supernatant to obtain the traditional Chinese medicine composition of the present invention.

[0122] Example 8

[0123] The main ingredients, Terminalia chebula and Glycyrrhiza uralensis, are in a mass ratio of 1:1.5.

[0124] The preparation method of this composition is as follows:

[0125] S1. After separately pulverizing Terminalia chebula and Glycyrrhiza uralensis, mix them in a mass ratio of 1:1.5, pass them through a 20-mesh sieve, and obtain powder raw materials;

[0126] S2. Add the powdered raw materials from S1 to deionized water at a mass-volume ratio of 1:5, adjust the pH value to 4.0 with 10% hydrochloric acid solution, and soak in a 45℃ water bath for 60 minutes.

[0127] S3. Add cellulase to the soaking solution of S2. The amount of cellulase added is 1.0% of the mass of the medicinal powder raw material; and carry out the enzymatic hydrolysis reaction in a water bath at 45°C for 90 minutes.

[0128] S4. Boil the enzymatic hydrolysis extract from S3 for 2 hours, filter, and set aside the filtrate. Add 5 times the weight of the herbal powder raw material in deionized water to the residue, boil for 2 hours, filter, combine the filtrates, and filter through a 1μm filter membrane to obtain the filtrate.

[0129] S5. Concentrate the filtrate from S4 under reduced pressure at 70℃ until the concentration of the drug solution is 0.3g crude drug / mL;

[0130] S6. Add 20 mg / mL of activated carbon to the concentrated solution of S5 according to the volume of the concentrated solution. Stir at 60°C for 3.5 h, remove the activated carbon, and obtain the supernatant. After drying, obtain the extract.

[0131] S7. Add one times the mass of deionized water and one times the mass of butylene glycol to the extract obtained in S6, stir to dissolve, let stand overnight, and take the supernatant to obtain the traditional Chinese medicine composition of the present invention.

[0132] Comparative Example 1

[0133] This comparative example provides a product made from Terminalia chebula alone. The amount of Terminalia chebula alone used in Comparative Example 1 is the sum of the amounts of Terminalia chebula and licorice in Example 2, and its preparation method is the same as in Example 1.

[0134] Comparative Example 2

[0135] This comparative example provides a product made from licorice alone. The amount of licorice alone used in Comparative Example 2 is the sum of the amounts of Terminalia chebula and licorice in Example 2, and its preparation method is the same as in Example 1.

[0136] Comparative Example 3

[0137] This comparative example provides a product made from the main raw materials Terminalia chebula and Glycyrrhiza uralensis in a mass ratio of 1:12, and its preparation method is the same as that in Example 1.

[0138] Comparative Example 4

[0139] This comparative example provides a product made from the main raw materials Terminalia chebula and Glycyrrhiza uralensis in a mass ratio of 12:1, and its preparation method is the same as that in Example 1.

[0140] The products of the embodiments and comparative examples are presented in the form of data tables, as follows:

[0141] Table 1. Raw material ratios used in the examples and comparative examples.

[0142]

[0143]

[0144] II. Verification of Experimental Results

[0145] Example of effect 1

[0146] This example demonstrates the antibacterial effects of the traditional Chinese medicine compositions prepared in Examples 1-8 and Comparative Examples 1-4 of the present invention.

[0147] 1. Experimental Methods

[0148] 1.1 Determination of inhibition zone

[0149] The diameter of the inhibition zone for each sample was determined using the agar perforation method. On a clean bench, sterilized tryptic soy agar (TSA) medium was cooled to approximately 60°C. 20 mL of the medium was poured into a petri dish and allowed to stand horizontally until completely solidified. An Oxford cup (6 mm inner diameter, 10 mm height) was placed on the surface of the agar medium, and a hole was gently punched under pressure. The medium was removed from the agar well using a sterile needle, and 0.1 mL of the test bacterial suspension was transferred to the plate and spread evenly using a spreader to prepare a bacterial plate. The prepared samples were then injected sequentially (80 μL per well). The petri dishes were then incubated at 37°C for 24 hours. The diameter of the inhibition zone was measured using calipers. A blank control group was prepared using 0.9% sterile sodium chloride solution.

[0150] 1.2 Method for determining the minimum inhibitory concentration (MIC)

[0151] The minimum inhibitory concentration (MIC) of each sample was determined using a two-fold serial dilution method. Solutions of different samples were serially diluted with TSB liquid medium to prepare solutions of specific concentrations. Indicator bacteria in the logarithmic growth phase were serially diluted with TSB liquid medium to 10⁻⁶. 5 For each CFU / mL bacterial suspension, 100 μL of the bacterial culture was added to wells 2-12 of a 96-well plate. An additional 100 μL of serial dilution was added to each well. The first well contained 100 μL of TSB liquid medium and 100 μL of bacterial suspension as a blank control. Each strain was tested three times. After mixing, the 96-well plate was incubated at 37°C for the specified time. Bacterial growth was observed. Each experiment was repeated three times, and the lowest concentration at which no colonies were observed in three trials was defined as the MIC.

[0152] 2. Experimental Results

[0153] The results of the inhibition zone assay are shown in Tables 1 and 2. Figures 1-5 The MIC measurement results are shown in Tables 3 and 4.

[0154] Table 2 Results of inhibition zone determination (n=3, )

[0155]

[0156] Note: Compared with Comparative Example 1 sample. # P<0.05, ## P<0.01, compared with the sample of Comparative Example 2, & P<0.05, && P<0.01, compared with comparative sample 3, * P<0.05, ** P<0.01, compared with comparative sample 4, $ P<0.05, $$ P<0.01.

[0157] Table 3. MIC experimental results (n=3)

[0158]

[0159] According to the results in Table 2, compared with Comparative Examples 1-4, the compositions of Examples 1-8 all showed significant antibacterial effects against Trichophyton mentagrophytes, Candida albicans, and Streptococcus mutans (P<0.01).

[0160] Similarly, the results in Table 3 show that the minimum inhibitory concentrations (MICs) of the compositions in Examples 1-8 were all lower than those of the comparative examples, indicating that these examples have significant antibacterial activity. Although the antibacterial effect of Comparative Example 1 was not significantly different from that of Example 5 in terms of Staphylococcus aureus and Porphyromonas gingivalis, this does not affect the positive evaluation of the overall antibacterial effect of the examples in this application. Combining the results in Tables 2 and 3, the examples showed significantly better performance than the comparative examples in terms of inhibition zone diameter and minimum inhibitory concentration in the antibacterial experiment.

[0161] The data above shows that the combination of Terminalia chebula and licorice has a synergistic effect compared to using either alone. When the weight ratio of Terminalia chebula and licorice is within a specific range, the combined effect is far superior to that of either alone, and also superior to cases outside that range. For example, in terms of antibacterial effect, when the ratio of Terminalia chebula to licorice is 2.5:1 (Example 3), its effect is better than that of the other sample examples.

[0162] Therefore, regarding the antibacterial effect, this invention screened an optimized formulation based on experimental results. Under the appropriate formulation, the combination of Terminalia chebula and Glycyrrhiza uralensis produced the best synergistic effect.

[0163] The synergistic effect of Terminalia chebula and licorice can be explained by their respective effective chemical components and their interaction mechanisms. Terminalia chebula is mainly rich in tannins and flavonoids, which are known for their strong antioxidant and antibacterial activities, especially in disrupting bacterial cell membranes and interfering with cell metabolism. The key active components of licorice are glycyrrhizic acid and glycyrrhizin flavonoids. The former inhibits bacterial enzyme activity or interferes with its protein synthesis pathways, while the latter directly inhibits bacterial growth through its anti-inflammatory and antibacterial properties.

[0164] When Terminalia chebula and licorice are combined in a specific ratio, their effective chemical components act on bacterial cells through different targets, forming a multi-target synergistic effect. The polyphenolic compounds of Terminalia chebula enhance the permeability of bacterial cell membranes, creating favorable conditions for the action of glycyrrhizic acid and glycyrrhizin flavonoids, thereby accelerating bacterial cell destruction. Simultaneously, glycyrrhizic acid further enhances the antibacterial effect of Terminalia chebula components by inhibiting bacterial metabolism and gene expression.

[0165] Therefore, the active ingredients of Terminalia chebula and licorice interact through multiple mechanisms, producing antibacterial effects far exceeding those of any single ingredient. This provides a solid theoretical foundation and promising application prospects for the development of antibacterial agents based on natural plants. This synergistic effect not only effectively enhances the antibacterial effect but also reduces the risk of bacteria developing drug resistance, possessing significant scientific and practical application value.

[0166] Example 2

[0167] This example demonstrates the antioxidant effects of the traditional Chinese medicine compositions prepared in Examples 1-8 and Comparative Examples 1-4 of the present invention.

[0168] 1. Experimental Methods

[0169] 1.1 Effect on DPPH radical scavenging

[0170] A certain amount of sample was weighed and dissolved in DMSO (analytical grade), and sonicated until fully dissolved. 80 μL of sample solutions of different concentrations and 80 μL of 1 mmol / L DPPH solution were added sequentially to a 96-well plate. After reacting at room temperature in the dark for 30 min, the absorbance was measured at 517 nm and recorded as A1. Three parallel measurements were performed, and the average value was taken. Vitamin C was used as a positive control.

[0171] The calculation formula is as follows:

[0172]

[0173] A2: Replace DPPH solution with the same volume of DMSO; A0: Replace the sample with the same volume of DMSO, and the procedure is the same as A1.

[0174] 1.2 ABTS + Effects of free radical scavenging

[0175] Accurately measure 1 mL each of 7.4 mmol / L ABTS solution and 2.6 mmol / L K₂S₂O₈ solution, mix well, and store at 4°C in the dark for 12–16 h to obtain ABTS stock solution. Then, mix this stock solution with distilled water at a specific ratio (to achieve an absorbance A = 0.700 ± 0.02) to obtain ABTS working solution. Add 10 μL of different concentrations of the sample and 200 μL of ABTS working solution sequentially to a 96-well plate. Incubate for 6 min at room temperature in the dark, and measure the absorbance at 734 nm, recording it as A1. Vitamin C was used as a positive control.

[0176] The calculation formula is as follows:

[0177]

[0178] A0: Replace the sample with the same volume of distilled water, and follow the same procedure as A1.

[0179] 2. Data Processing

[0180] Based on the linear fitting curves of DPPH and ABTS radical scavenging of different sample solutions, regression analysis using IBM SPSS Statistics 26 was used to calculate the amount of sample solution added when the original mass concentrations of DPPH and ABTS were reduced to 50% (steady state), which is the half-inhibitory amount (IC50). 50 According to half-inhibition level (IC50) 50 The size of the IC50 value can determine the strength of the sample solution's ability to scavenge free radicals. 50 The smaller the value, the stronger the ability to scavenge free radicals.

[0181] 3. Experimental Results

[0182] 3.1 In vitro antioxidant (DPPH, ABTS) performance analysis

[0183] The results are as follows Figures 6-7 As shown, in Example 6, the DPPH radical scavenging rate reached 50% at a concentration of 0.24 mg / mL, while the DPPH radical scavenging rate in the comparative samples was significantly less than 50%. The scavenging rate reached its maximum at approximately 3.125 mg / mL, stabilizing at around 94%, which was higher than the positive control, vitamin C. In scavenging ABTS radicals, the inhibition rate of Example 6 at a concentration of approximately 0.39 mg / mL was stable at around 94%, higher than the positive control. Other comparative samples also showed an increased antioxidant effect with increasing sample concentration. However, the in vitro antioxidant effect of the comparative samples was not as good as that of Example 6.

[0184] 3.2 Analysis of in vitro antioxidant assay results

[0185] Table 4. Results of in vitro antioxidant activity determination for Examples 1-8 and Comparative Examples 1-4 (n=3, )

[0186]

[0187] Note: Compared with comparative sample 1. & P<0.05, && P<0.01, compared with comparative sample 2, # P<0.05, ## P<0.01, compared with comparative sample 3, * P<0.05, ** P<0.01, compared with comparative sample 4, $ P<0.05, $$ P<0.01.

[0188] Based on the results in Table 4, it is evident that the samples from Examples 1-8 significantly outperformed the comparative samples in their ability to scavenge DPPH and ABTS free radicals (P<0.01). Among them, the sample from Example 6 (with a mass ratio of Terminalia chebula to licorice of 5:1) showed the best effect in scavenging DPPH and ABTS free radicals. Overall, the samples from the examples exhibited superior in vitro antioxidant effects, far exceeding those of the comparative samples.

[0189] The data above shows that the combination of Terminalia chebula and licorice has a synergistic effect compared to using either alone. When the weight ratio of Terminalia chebula and licorice is within a specific range, the combined effect is far superior to that of either ingredient alone, and also superior to cases outside that range. For example, in terms of antioxidant effect, when the ratio of Terminalia chebula to licorice is 5:1, its effect is better than the samples in other examples.

[0190] The synergistic effect of Terminalia chebula and licorice in antioxidant efficacy mainly stems from the complementarity of their respective active ingredients. The tannins and flavonoids of Terminalia chebula rapidly scavenge free radicals, while the glycyrrhizic acid and glycyrrhizin of licorice prolong the duration of antioxidant effects by inhibiting free radical generation and enhancing the activity of cellular antioxidant enzymes. When combined in a specific ratio, they not only significantly improve the scavenging capacity of DPPH and ABTS free radicals but also enhance the persistence and stability of the overall antioxidant effect. Experiments have verified that the optimized ratio of this combination effectively improves the antioxidant effect, demonstrating the scientific value and practical application potential of plant-based compound formulations in the field of antioxidants. Therefore, based on experimental results, this invention has screened an optimized ratio scheme for antioxidant effects. Under an appropriate ratio, the combination of Terminalia chebula and licorice produces the optimal synergistic effect.

[0191] Example 3

[0192] This example demonstrates the anti-inflammatory effects of the traditional Chinese medicine compositions prepared in Examples 1-8 and Comparative Examples 1-4 of this invention.

[0193] 1. Experimental Methods

[0194] Mouse monocyte / macrophage RAW264.7 cells were purchased from Wuhan Pronosei Biotechnology Co., Ltd. The RAW264.7 cell line was seeded in DMEM medium containing 10% FBS and 1% penicillin / streptomycin antibiotics. In the anti-inflammatory assay, the RAW264.7 cell line was seeded at a concentration of 1 × 10⁻⁶ cells / cm². 5RAW264.7 cells were seeded into 24-well plates (cells / well), with a blank control group, an LPS group, and various drug-treated groups, as well as a positive control group (dexamethasone). After culturing in a 5% CO2, 37°C incubator for 12 hours, the cells were pre-treated with samples from the embodiments of this invention and comparative samples (100 μg / mL) for 12 hours. Then, RAW264.7 cells were induced with lipopolysaccharide (LPS, 1 μg / mL) for 24 hours. After induction, the cell culture supernatant was collected, and the production of inflammatory factors was detected using an ELISA kit.

[0195] 2. Statistical methods

[0196] The experimental data were statistically analyzed using SPSS 22.0 software. The statistics were expressed as mean ± standard deviation (X ± SD), and one-way ANOVA was used for comparisons between groups.

[0197] 3. Experimental Results

[0198] The results of the levels of pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 are shown in Table 5.

[0199] Table 5. Results of Inflammatory Markers

[0200]

[0201] Note: Compared with the blank group, ## P < 0.01. Compared with the model group, ** P < 0.01. Compared with Comparative Example 1, $ P<0.05, $$ P<0.01. Compared with Comparative Example 2, & P<0.05, && P<0.01, compared with comparative sample 3, Compared with comparative sample 4, § P<0.05, §§ P<0.01.

[0202] Table 5 shows that LPS significantly stimulated the release of inflammatory factors in RAW264.7 cells, with a significant increase in the levels of inflammatory factors (such as TNF-α, IL-1β, and IL-6) compared to the control group (P<0.01). In each sample treatment group, the samples from Examples 1-8 and Comparative Examples 1-4 significantly reduced the levels of these inflammatory factors, exhibiting significant anti-inflammatory effects compared to the model group (P<0.01). Notably, the samples from each example were superior to the comparative examples in reducing inflammatory factor levels (P<0.01), indicating that the anti-inflammatory effect of this traditional Chinese medicine composition was significantly better than that of a single herbal formula.

[0203] The data above shows that the combination of Terminalia chebula and licorice has a synergistic effect compared to using either alone. When the weight ratio of Terminalia chebula and licorice is within a specific range, the combined effect is far superior to that of either alone, and also superior to cases outside that range. For example, in terms of anti-inflammatory effect, when the ratio of Terminalia chebula to licorice is 2.5:1 (Example 3), its effect is better than that of the other sample examples.

[0204] The synergistic effect of Terminalia chebula and Glycyrrhiza uralensis in anti-inflammatory efficacy stems from the interaction of their respective anti-inflammatory components through different mechanisms. The polyphenolic compounds in Terminalia chebula effectively inhibit the activation of early inflammatory signals and reduce the release of pro-inflammatory factors; while glycyrrhizic acid in Glycyrrhiza uralensis further slows the spread and persistence of the inflammatory response by inhibiting the NF-κB and MAPK pathways. When the two are combined in a specific ratio, they exhibit significantly better anti-inflammatory effects than either alone, demonstrating a synergistic effect in multi-target inflammation suppression. This optimized ratio not only enhances the anti-inflammatory effect but also provides theoretical support and application prospects for the development of natural anti-inflammatory drugs.

[0205] Example of effect 4

[0206] This example provides the whitening effect (evaluation of tyrosinase inhibition effect) of the traditional Chinese medicine compositions prepared in Examples 1-8 and Comparative Examples 1-4 of the present invention.

[0207] 1. Experimental Methods

[0208] First, prepare 368 U / mL tyrosinase solution and 1.0 mg / mL tyrosine solution, with a total reaction volume of 100 μL. The composition of the reaction solution is shown in Table 6. Using a pipette, accurately pipette the substrate, solvent, and reagent from solutions a, b, c, and d into the corresponding wells of a 96-well plate, performing three replicates for each concentration. Mix thoroughly and incubate at 25°C for 10 min. After adding 368 U / mL tyrosinase solution, incubate the entire 96-well plate at 37°C for 20 min. Immediately place the plate in a microplate reader and measure the absorbance (A) at 475 nm, labeling them Aa, Ab, Ac, and Ad respectively. Inhibition rate = [1 - (Ac - Ad) / (Aa - Ab)] × 100%. α-Arbutin was used as a positive control.

[0209] Aa: Absorbance at 475 nm of the mixture containing tyrosinase and without sample.

[0210] Ab: Absorbance at 475 nm of the mixture without sample and tyrosinase.

[0211] Ac: Absorbance of the mixture of sample and tyrosinase at 475 nm

[0212] Ad: Absorbance at 475 nm of the mixture with sample but without tyrosinase.

[0213] Table 6 Tyrosinase Inhibition Rate (IC50) 50

[0214]

[0215] Note: Compared with comparative sample 1. & P<0.05, && P<0.01, compared with comparative sample 2, # P<0.05, ## P<0.01, compared with comparative sample 3, * P<0.05, ** P<0.01, compared with comparative sample 4, $ P<0.05, $$ P<0.01.

[0216] As shown in Table 6, the samples from Examples 1-8 exhibited significantly better inhibitory effects on tyrosinase than those from Comparative Examples 1-4 (P<0.01). Compared to the comparative samples, the sample examples demonstrated stronger tyrosinase inhibitory activity, as evidenced by their lower IC50 values. This indicates that the sample examples possess superior tyrosinase inhibitory effects. Therefore, the sample examples can be added as cosmetic ingredients to effectively enhance the whitening efficacy of the final product. In contrast, while the sample examples from Comparative Examples 1-2 showed some inhibitory effects on tyrosinase, their IC50 values ​​were relatively high. Applying them to the final product would increase costs and result in poor whitening effects.

[0217] The data above shows that the combination of Terminalia chebula and licorice has a synergistic effect compared to using either alone. When the weight ratio of Terminalia chebula and licorice is within a specific range, the combined effect is far superior to that of either ingredient alone, and also superior to cases outside that range. For example, in terms of whitening effect, when the ratio of Terminalia chebula to licorice is 1:5 (Example 5), the effect is better than that of the other sample examples.

[0218] Example 5

[0219] This example provides an evaluation test of the anti-glycation effect of the traditional Chinese medicine compositions prepared in Examples 1-8 and Comparative Examples 1-4 of this invention.

[0220] 1. Experimental Methods

[0221] 1.1 Establishment of an in vitro protein AGE formation system

[0222] A solution of bovine serum albumin (BSA) and 0.5 mol / L glucose with a final concentration of 20 mg / mL was prepared using phosphate buffer (pH 7.4) as the matrix. The solutions were mixed 1:1, sterilized by filtration through a 0.45 μm filter, and aliquoted into 1.5 mL conical centrifuge tubes. The tubes were incubated at 37°C and 60°C under aerobic and light-protected conditions for 24 h, respectively. The OD value of the formed AGE was measured using a microplate reader (detection wavelength 370 nm, reference wavelength 440 nm).

[0223] 1.2 Assay for in vitro inhibition of AGEs formation

[0224] A BSA-glucose model reaction system was used. Examples 3, 1, and 2 were sequentially mixed with bovine serum albumin (BSA)-glucose model solution at a 1:1 ratio to form sample groups (Group A). ​​Examples 3, 1, and 2 were sequentially mixed with phosphate buffer at a 1:1 ratio to form sample groups (Group B). The model solution was mixed with phosphate buffer at a 1:1 ratio to form the negative control group (Group C). Phosphate buffer solution was used as the blank control group (Group D). Aminoguanidine was used as the positive control. The reaction was incubated at 60°C in the dark for 5 hours, then terminated at 4°C for 10 minutes. Fluorescence intensity was detected using a multi-mode microplate reader at an excitation wavelength of 370 nm and an emission wavelength of 440 nm. The AGEs inhibition rate was calculated using the following formula.

[0225]

[0226] Table 7 Anti-glycation effect

[0227]

[0228] 2. Experimental Results

[0229] According to the data in Table 7, the traditional Chinese medicine compositions obtained in Examples 1-8 all exhibited good anti-glycation effects. Compared with the comparative examples, the inhibition rates of AGEs by the traditional Chinese medicine compositions obtained in the examples were all superior, indicating that the traditional Chinese medicine compositions had excellent effects in inhibiting AGE formation. This suggests that in the compositions studied, the combination of two components can effectively reduce the level of non-enzymatic glycation end products (AGEs) and lower the expression level of advanced glycation end products (CML), exhibiting significant anti-glycation effects, thereby achieving the effects of skin whitening and brightening, and delaying aging.

[0230] The data above shows that the combination of Terminalia chebula and licorice has a synergistic effect compared to using either alone. When the weight ratio of Terminalia chebula and licorice is within a specific range, the combined effect is far superior to that of either alone, and also superior to cases outside that range. For example, in terms of anti-glycation effect, when the ratio of Terminalia chebula to licorice is 7:1 (Example 7), its effect is better than the samples in other examples.

[0231] Example 6

[0232] This example demonstrates the antipruritic effect of the traditional Chinese medicine compositions prepared in Examples 1-8 and Comparative Examples 1-4 of this invention.

[0233] 1. Experimental Methods

[0234] Mouse mast cell line MC / 9 was purchased from the American Type Organisms Collection (ATCC). MC / 9 cells were seeded in IMEM medium containing 10% FBS and 1% penicillin / streptomycin antibiotics. Cells were cultured at 37°C in a 5% CO2 incubator, and cells in the logarithmic growth phase were used for experiments. In the histamine production assay, cells were seeded in 24-well plates at a density of 1 × 10⁶ cells / well. 5 Cells were cultured at a concentration of 100 μg / mL for 12 hours. Then, they were pre-treated and protected for 12 hours with samples from Example 1, Example 2, Comparative Example 1, Comparative Example 2, Comparative Example 3, and Comparative Example 4 (100 μg / mL). Following this, MC / 9 cells were induced for 24 hours with propylene glycol methyl ether acetate (PMA, 50 nM) and calcium iontophoresis agent A23187 (1 μM). After induction, the cell culture supernatant was collected, and histamine production was detected using an ELISA kit.

[0235] 2. Statistical methods

[0236] The experimental data were statistically analyzed using SPSS 22.0 software, and the statistical measures were expressed as mean ± standard deviation. This indicates that one-way ANOVA was used for comparisons between groups.

[0237] 3. Experimental Results

[0238] Table 8. Experimental results on inhibition of histamine production (n=3, )

[0239]

[0240]

[0241] Note: Compared with the blank group, ## P<0.01. Compared with the PMA / A23187 group, & P<0.05,&& P < 0.01. Compared with Comparative Example 1, * P<0.05, ** P<0.01. Compared with Comparative Example 2, $ P<0.05, $$ P<0.01, compared with comparative sample 3, Compared with comparative sample 4, § P<0.05, §§ P<0.01.

[0242] Table 8 shows that PMA / A23187 stimulation significantly promoted histamine release in MC / 9 cells, with a significantly increased histamine content compared to the control group (P<0.01). In all sample treatment groups, the herbal compositions of Examples 1-8 significantly inhibited histamine release, demonstrating good anti-allergic effects (P<0.01). Furthermore, all examples showed better results than comparative examples 1-4, exhibiting more significant inhibitory effects.

[0243] The data above shows that the combination of Terminalia chebula and licorice has a synergistic effect compared to using either alone. When the weight ratio of Terminalia chebula and licorice is within a specific range, the combined effect is far superior to that of either alone, and also superior to cases outside that range. For example, in terms of antipruritic effect, when the ratio of Terminalia chebula to licorice is 10:1 (Example 2), its effect is better than that of the other sample examples.

[0244] Example 7

[0245] In addition to the synergistic effect of the aforementioned efficacy, the combination of Terminalia chebula and Glycyrrhiza uralensis can also improve the stability of the active ingredients in Terminalia chebula at different pH values. This example provides a stability test of the total phenolic, gallic acid, and glycyrrhizic acid content of the sample of Example 3 of the present invention and Terminalia chebula alone (Comparative Example 1) and Glycyrrhiza uralensis alone (Comparative Example 2) under different pH conditions.

[0246] 1. Experimental Methods

[0247] The Elite Hypersil BDS C18 column (4.6 × 250 mm, 5 μm) was used. High-performance liquid chromatography (HPLC) detection conditions were as follows: 0.1% phosphoric acid aqueous solution-chromatographic grade methanol was used as the mobile phase, with gradient elution: 0–5 min, 5%–10% methanol; 5–10 min, 10%–11% methanol; 10–15 min, 11%–15% methanol; 15–20 min, 15–25% methanol; 20–30 min, 25%–40% methanol; 30–40 min, 40%–45% methanol; 40–50 min, 45%–70% methanol; 50–60 min, 70%–80% methanol; column temperature was 25℃, detection wavelength was 254 nm, flow rate was 1 mL / min, and injection volume was 20 μL.

[0248] The total phenol content of Terminalia chebula was determined using the Folin-Ciocalteu colorimetric method, with gallic acid from Terminalia chebula as a standard.

[0249] 2. Experimental Results

[0250] Table 9. Experimental results of changes in total phenols and gallic acid content of Terminalia chebula extract at different pH values ​​in Example 3.

[0251]

[0252] Table 10. Experimental results of changes in total phenols and gallic acid content of licorice extract at different pH levels in Example 3.

[0253]

[0254] Table 9-10 shows the experimental results. Within the pH range of 5.0–7.5, the total phenol and gallic acid contents in Example 3 fluctuated relatively little. At pH 5.0, the gallic acid content in Example 3 was 10.68 mg / mL, and the total phenol content was 50.36 mg / mL; at pH 7.5, the gallic acid content was 10.38 mg / mL, and the total phenol content was 49.01 mg / mL. These results indicate that increasing pH has little effect on the total phenol and gallic acid contents in Example 3.

[0255] In the Terminalia chebula single-ingredient sample (Comparative Example 1), within this pH range, the contents of total phenols and gallic acid decreased significantly with increasing pH. At pH 5.0, the gallic acid content in the Terminalia chebula single-ingredient sample was 10.29 mg / mL, and the total phenol content was 47.21 mg / mL. At pH 7.5, the gallic acid content was 7.07 mg / mL, and the total phenol content was 32.11 mg / mL. These results indicate that with increasing pH, the stability of gallic acid and total phenols in the Terminalia chebula single-ingredient sample was affected, and their contents decreased significantly.

[0256] Within the pH range of 5.0 to 7.5, the glycyrrhizic acid content in Example 3 and the licorice extract (Comparative Example 2) showed little change, indicating that pH fluctuations within this range have little impact on the glycyrrhizic acid content.

[0257] The above results indicate that when Terminalia chebula and licorice are used in combination, the total phenolic components exhibit better stability under different pH conditions. This may be related to the interactions between the components in the formulation, particularly those in licorice. These interactions may enhance the stability of the active ingredients, thereby maintaining high bioactivity under various environmental conditions. This finding is of great significance for developing products with good stability.

[0258] These experimental results demonstrate that the traditional Chinese medicine composition of this invention exhibits a significant synergistic effect through scientific formulation. The antibacterial, anti-inflammatory, antioxidant, whitening, antipruritic, and anti-glycation effects of all embodiments are superior to those of the comparative sample group. This indicates that the present invention optimizes the relevant properties of the traditional Chinese medicine composition during formulation and preparation, further proving the importance of the preparation method of this invention in enhancing the antibacterial, anti-inflammatory, antioxidant, whitening, antipruritic, and anti-glycation effects of the traditional Chinese medicine composition.

[0259] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions or improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. Use of a composition prepared from main materials of Terminalia chebula and Glycyrrhiza glabra in the preparation of antibacterial and anti-inflammatory products, characterized in that, The main materials include Terminalia chebula and Glycyrrhiza uralensis, and the mass ratio of Terminalia chebula to Glycyrrhiza uralensis is 2.5:1; The preparation method of the bacteriostatic and anti-inflammatory product is as follows: S1, Terminalia chebula and Glycyrrhiza uralensis are respectively crushed, mixed according to the mass ratio of 2.5:1, and passed through a 20-mesh sieve to obtain powder raw materials; S2, the powder raw materials in S1 are added into deionized water according to the mass-volume ratio of 1:5, and 10% hydrochloric acid solution is used to adjust the pH value to 4.0, and then soaked in a water bath at 45°C for 60 minutes; S3, cellulase is added into the soaking solution in S2, and the addition amount of cellulase is 1.0% of the mass of the medicinal material powder raw materials; and enzymatic reaction is carried out in a water bath at 45°C for 90 minutes; S4, the enzymatic extraction liquid in S3 is decocted and extracted for 2 hours, filtered, and the filtrate is prepared for use; and the filter residue is added into deionized water with a mass of 5 times of the medicinal material powder raw materials, decocted and extracted for 2 hours, and filtered; The two filtrates are combined, filtered through a filter membrane with a pore size of 1 μm to obtain a filtrate; S5, the filtrate in S4 is concentrated under reduced pressure at 70°C, and the concentration of the medicinal liquid is 0.3 g of crude drug per mL; S6, 20 mg / mL of activated carbon is added into the concentrated liquid in S5 according to the volume of the concentrated liquid, stirred at 60°C for 3.5 hours, and then the activated carbon is removed to obtain a supernatant, which is dried to obtain an extract; S7, the extract obtained in S6 is added into deionized water with a mass of 1 times of the medicinal material raw materials and 1,2-propanediol with a mass of 1 times of the medicinal material raw materials, stirred and dissolved, and then left to stand overnight, and the supernatant is obtained, which is the composition.

2. Use of a composition prepared from main ingredients of Terminalia chebula and Glycyrrhiza glabra in the preparation of antioxidant products, characterized in that, The main materials include Terminalia chebula and Glycyrrhiza uralensis, and the mass ratio of Terminalia chebula to Glycyrrhiza uralensis is 5:1; The preparation method of the antioxidant product is as follows: S1, Terminalia chebula and Glycyrrhiza uralensis are respectively crushed, mixed according to the mass ratio of 5:1, and passed through a 20-mesh sieve to obtain powder raw materials; S2, the powder raw materials in S1 are added into deionized water according to the mass-volume ratio of 1:20, and 10% hydrochloric acid solution is used to adjust the pH value to 4.0, and then soaked in a water bath at 50°C for 60 minutes; S3, cellulase is added into the soaking solution in S2, and the addition amount of cellulase is 0.6% of the mass of the medicinal material powder raw materials; and enzymatic reaction is carried out in a water bath at 50°C for 90 minutes; S4, the enzymatic extraction liquid in S3 is decocted and extracted for 1.5 hours, filtered, and the filtrate is prepared for use; and the filter residue is added into deionized water with a mass of 20 times of the medicinal material powder raw materials, decocted and extracted for 1.5 hours, and filtered, and the filtrates are combined and filtered through a filter membrane with a pore size of 5 μm to obtain a filtrate; S5, the filtrate in S4 is concentrated under reduced pressure at 70°C, and the concentration of the medicinal liquid is 0.5 g of crude drug per mL; S6, 20 mg / mL of activated carbon is added into the concentrated liquid in S5 according to the volume of the concentrated liquid, stirred at 60°C for 3 hours, and then the activated carbon is removed to obtain a supernatant, which is dried to obtain an extract; S7, the extract obtained in S6 is added into deionized water with a mass of 1.5 times of the medicinal material and 1,2-propanediol with a mass of 1.5 times of the medicinal material, stirred and dissolved, and then left to stand overnight, and the supernatant is obtained, which is the composition.

3. Use of a composition prepared from main materials of Terminalia chebula and Glycyrrhiza glabra in the preparation of whitening products, characterized in that, The main materials include Terminalia chebula and Glycyrrhiza uralensis, and the mass ratio of Terminalia chebula to Glycyrrhiza uralensis is 1:5; The preparation method of the whitening product is as follows: S1, Terminalia chebula and Glycyrrhiza uralensis are respectively crushed, mixed according to the mass ratio of 1:5, and passed through a 20-mesh sieve to obtain powder raw materials; S2, the powder raw material in S1 is added into deionized water according to the mass-volume ratio of 1:20, and 10% hydrochloric acid solution is used to adjust the pH value to 4.5, and then the mixture is soaked in a water bath at 60℃ for 70 minutes; S3, cellulase is added into the soaking solution of S2, and the addition amount of cellulase is 0.3% of the mass of the medicinal material powder raw material; and the enzymolysis reaction is carried out in a water bath at 55℃ for 70 minutes; S4, the enzymolysis extraction solution of S3 is decocted and extracted for 2.5 hours, and then filtered, and the filtrate is reserved; and the filter residue is added with deionized water in an amount of 20 times of the mass of the medicinal material powder raw material, and then decocted and extracted for 2.5 hours, and then filtered, and the filtrate is combined, and then filtered through a filter membrane with a pore size of 0.45 μm to obtain a filtrate; S5, the filtrate of S4 is concentrated under reduced pressure at 75℃ until the concentration of the medicinal liquid is 0.45 g of crude drug per mL; S6, 25 mg / mL of activated carbon is added into the concentrated solution of S5 according to the volume of the concentrated solution, and then stirred at 75℃ for 5 hours, and then the activated carbon is removed to obtain a supernatant, and then the supernatant is dried to obtain an extract; S7, the extract obtained in S6 is added with deionized water in an amount of 1 times of the mass of the medicinal material raw material and 1,3-propanediol in an amount of 1 times of the mass of the medicinal material raw material, and then stirred and dissolved, and then left to stand overnight, and then the supernatant is taken to obtain the composition.

4. Use of a composition prepared from main ingredients of guggal and liquorice in the preparation of anti-glycation products, characterized in that, The main materials include Terminalia chebula and Glycyrrhiza uralensis, and the mass ratio of Terminalia chebula to Glycyrrhiza uralensis is 7:1; The preparation method of the anti-glycation product is as follows: S1, Terminalia chebula and Glycyrrhiza uralensis are respectively crushed, mixed according to the mass ratio of 7:1, and then passed through a 10-mesh sieve to obtain a powder raw material; S2, the powder raw material in S1 is added into deionized water according to the mass-volume ratio of 1:10, and 10% hydrochloric acid solution is used to adjust the pH value to 5.0, and then the mixture is soaked in a water bath at 65℃ for 30 minutes; S3, cellulase is added into the soaking solution of S2, and the addition amount of cellulase is 0.2% of the mass of the medicinal material powder raw material; and the enzymolysis reaction is carried out in a water bath at 65℃ for 30 minutes; S4, the enzymolysis extraction solution of S3 is decocted and extracted for 1 hour, and then filtered, and the filtrate is reserved; and the filter residue is added with deionized water in an amount of 10 times of the mass of the medicinal material powder raw material, and then decocted and extracted for 1 hour, and then filtered, and the filtrate is combined, and then filtered through a filter membrane with a pore size of 10 μm to obtain a filtrate; S5, the filtrate of S4 is concentrated under reduced pressure at 60℃ until the concentration of the medicinal liquid is 0.2 g of crude drug per mL; S6, 50 mg / mL of activated carbon is added into the concentrated solution of S5 according to the volume of the concentrated solution, and then stirred at 80℃ for 2 hours, and then the activated carbon is removed to obtain a supernatant, and then the supernatant is dried to obtain an extract; S7, the extract obtained in S6 is added with deionized water in an amount of 2 times of the mass of the medicinal material raw material and 1,3-propanediol in an amount of 2 times of the mass of the medicinal material raw material, and then stirred and dissolved, and then left to stand overnight, and then the supernatant is taken to obtain the composition.

5. Use of a composition prepared from main ingredients of gugal and liquorice in the preparation of an anti-itch product, characterized in that, The main materials include Terminalia chebula and Glycyrrhiza uralensis, and the mass ratio of Terminalia chebula to Glycyrrhiza uralensis is 10:1; The preparation method of the anti-glycation product is as follows: S1, Terminalia chebula and Glycyrrhiza uralensis are respectively crushed, mixed according to the mass ratio of 10:1, and then passed through a 10-mesh sieve to obtain a powder raw material; S2, the powder raw material in S1 is added into deionized water according to the mass-volume ratio of 1:10, and 10% hydrochloric acid solution is used to adjust the pH value to 5.0, and then the mixture is soaked in a water bath at 65℃ for 30 minutes; S3, cellulase is added to the soaking solution of S2, and the amount of cellulase added is 0.2% of the mass of the medicinal powder raw material; enzymatic reaction is carried out in a water bath at 65℃ for 30 minutes; S4, the enzymatic extraction solution of S3 is decocted and extracted for 1 hour, filtered, and the filtrate is reserved; the filter residue is added with deionized water in an amount of 10 times the mass of the medicinal powder raw material, decocted and extracted for 1 hour, filtered, and the filtrate is combined; the combined filtrate is filtered through a filter membrane with a pore size of 10 μm to obtain a filtrate; S5, the filtrate of S4 is concentrated under reduced pressure at 60℃ until the concentration of the medicinal liquid is 0.2g crude drug / mL; S6, to the concentrated solution of S5, 50mg / mL of activated carbon is added according to the volume of the concentrated solution; after stirring at 80℃ for 2 hours, the activated carbon is removed to obtain a supernatant, which is dried to obtain an extract; S7, the extract obtained in S6 is added with deionized water in an amount of 2 times the mass of the medicinal raw material and 1,3-propanediol in an amount of 2 times the mass of the medicinal raw material, stirred and dissolved, and left to stand overnight; the supernatant is taken to obtain the composition.

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