Chinese medicine composition with bacteriostatic, anti-inflammatory, antioxidant and whitening effects, preparation method and application thereof

Abstract

This invention belongs to the field of composition technology, and specifically relates to a traditional Chinese medicine composition with antibacterial, anti-inflammatory, antioxidant, and whitening effects, its preparation method, and its application. The composition of this application comprises the following raw materials in parts by weight: 1-5 parts Rhodiola rosea; 5-10 parts Glycyrrhiza uralensis; and 1-5 parts Phyllanthus emblica. The composition is obtained by pulverizing Rhodiola rosea, Glycyrrhiza uralensis, and Phyllanthus emblica into powder, adding them to an extraction solvent, and then extracting. Compared with single-purpose compositions, the composition of this application simultaneously possesses antibacterial, anti-inflammatory, antioxidant, and whitening effects, thus simultaneously satisfying the above four efficacy requirements of the product.

Description

Technical Field

[0001] This invention belongs to the field of composition technology, and in particular relates to a traditional Chinese medicine composition that simultaneously has antibacterial, anti-inflammatory, antioxidant and whitening effects, as well as its preparation method and application. Background Technology

[0002] With social development, continuous improvement in lifestyles, and constant changes in the living environment, people's health awareness is also constantly increasing. Therefore, multi-functional daily chemical products have become a new demand in people's daily consumption. The skin is the largest organ of the human body, undertaking multiple functions such as protecting the body, perspiration, and sensing hot, cold, and pressure. Under normal circumstances, numerous microorganisms exist on the surface of human skin, maintaining a microecological balance with the human body, thereby safeguarding the body's health. Once this balance is disrupted, it can cause skin inflammation, redness, itching, and even more serious diseases. Therefore, effectively inhibiting pathogenic bacteria on the skin while maintaining microecological balance is a basic requirement for skin cleansing and skincare products. Furthermore, if these products also possess anti-inflammatory, antioxidant, and whitening effects, it will expand the scope of application and increase consumer comfort.

[0003] Traditional Chinese medicine (TCM) and its compound preparations are primarily used internally, with external application as a secondary method. Therefore, water is the most commonly used extraction solvent, with only a very few plaster preparations using plant oils. What active effects would be produced by using cosmetic base oils as solvents to extract TCM? Due to a lack of necessary research data, related reports are scarce. Therefore, it is necessary to provide a TCM skincare ingredient using base oils as solvents. Through extensive experiments, this invention has determined that using base oils as solvents, combined with Rhodiola rosea, licorice, and Phyllanthus emblica, exhibits significant antibacterial, anti-inflammatory, antioxidant, and whitening effects, making it suitable for use in finished cosmetic products. Summary of the Invention

[0004] To address the aforementioned technical problems, this invention provides a traditional Chinese medicine composition that simultaneously possesses antibacterial, anti-inflammatory, antioxidant, and whitening effects. This composition offers advantages such as antibacterial, anti-inflammatory, antioxidant, and whitening properties, overcoming the shortcomings of the aforementioned raw materials having only one function. Furthermore, the composition of this invention uses a base oil as a solvent, exhibiting strong activity and a light color, making it particularly suitable for skincare products such as lotions, creams, makeup removers, and serums.

[0005] The composition of the present invention comprises the following raw materials in parts by weight: 1 to 5 parts Rhodiola rosea; 5 to 10 parts Glycyrrhiza uralensis; 1 to 5 parts Phyllanthus emblica.

[0006] The preparation method of the above-mentioned traditional Chinese medicine composition includes the following steps:

[0007] (1) Pulverize 1-5 parts of Rhodiola rosea, 5-10 parts of licorice and 1-5 parts of Phyllanthus emblica, and pass them through a 10-20 mesh sieve to obtain powder raw materials;

[0008] (2) Add the powdered raw material from (1) to the extraction solvent at a mass-volume ratio of 1g:(2-20)mL, extract at 50℃-180℃ for 0.5h-3.0h, filter through a 60-100 mesh filter cloth, discard the residue, collect the filtrate, and obtain the filtrate; the extraction solvent is selected from at least one of the following: coconut oil alcohol-caprylate / caprylate, propyl heptyl caprylate, phytosterol / octyldodecyl lauroyl glutamate, triolein, meadowfoam seed oil, squalane, coconut oil, olive oil, sweet almond oil, grape seed oil, jojoba oil, rosehip oil, sesame oil, wheat germ oil, camellia seed oil, caprylic acid / caprylic acid triglyceride;

[0009] (3) Filter the filtrate obtained in (2) through a filter plate of 2μm to 10μm until the filtrate is clear, and obtain the clear filtrate and the traditional Chinese medicine composition.

[0010] The above-mentioned traditional Chinese medicine composition is used in the preparation of antibacterial products, anti-inflammatory products, antioxidant products, and whitening products.

[0011] Rhodiola rosea has a bitter taste and cold properties, and is believed to have the effects of clearing heat and drying dampness, and strengthening the body. Modern research shows that Rhodiola rosea contains abundant chemical components, including flavonoids, organic acids, and trace elements. Its main active ingredient is rhodioloside, and it also contains tyrosol, quercetin, and other compounds. Pharmacological activity studies have shown that Rhodiola rosea has various medicinal effects, including anti-hypoxia, anti-fatigue, antiviral, anti-radiation, and anti-aging properties.

[0012] Licorice has a sweet and neutral flavor and functions to tonify the spleen and replenish qi, clear heat and detoxify, relieve spasms and pain, and harmonize the effects of 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 to alleviate the toxicity of other drugs. In addition to glycyrrhizic acid and glycyrrhizin, licorice also contains various flavonoids such as glycyrrhizin, isoglycyrrhizin, and polysaccharides. Modern pharmacological studies have shown that licorice has various effects, including anti-inflammatory, antiviral, antioxidant, immunomodulatory, anti-atherosclerotic, and anti-tumor properties.

[0013] Phyllanthus emblica has a sour, sweet, and astringent taste, and is cool in nature. It possesses the effects of quenching thirst, clearing heat and detoxifying, moistening the lungs and relieving cough, and nourishing and promoting tissue regeneration. Modern research shows that Phyllanthus emblica contains various chemical components, including polyphenols, organic acids, tannins, vitamin C, and trace elements. Its main active ingredients are ellagic acid, gallic acid, and vitamin C. Pharmacological studies show that Phyllanthus emblica has powerful antioxidant, anti-inflammatory, antiviral, anti-aging, immunomodulatory, and hepatoprotective effects.

[0014] Based on the characteristics of traditional Chinese medicine compound formulas with "multiple components, multiple targets, and multiple pathways" and combined with the comprehensive needs of skin cleansing and care products, this invention focuses on multiple effects such as antibacterial, anti-inflammatory, antioxidant, and whitening. It combines Rhodiola rosea, licorice, and Phyllanthus emblica into a compound formula, which has a significant synergistic effect, can take into account multiple functions, and has high safety. It can be used as a functional additive in daily skin chemical products.

[0015] In cosmetics, base oils are essential ingredients in formulation. They are typically oils extracted from natural plants or animals, but can also be obtained synthetically. Commonly used base oils in cosmetics include plant oils (such as jojoba oil, sweet almond oil, and olive oil), animal oils (such as lanolin, squalane, and beeswax), and synthetic oils (such as caprylic / capric triglycerides, isononyl isononanoate, and isopropyl palmitate). The main functions of base oils in cosmetics include moisturizing, nourishing the skin, dissolving active ingredients, promoting ingredient absorption, and improving the stability of the formulation. Therefore, base oils are widely used in cosmetic products such as lotions, creams, serums, and lipsticks. As a raw material, base oils are also important solvents, used for the extraction and dissolution of other ingredients, thereby enhancing the skincare efficacy of the product.

[0016] The beneficial effects of this invention are as follows:

[0017] (1) Compared with single-purpose compositions, the compositions in this application have antibacterial, anti-inflammatory, antioxidant and whitening effects, and can simultaneously satisfy the above four effects of the product.

[0018] (2) The results showed that the average value of the inhibition zone of Staphylococcus aureus, Propionibacterium acnes and Trichophyton mentagrophytes was greater than 18 mm, indicating that the composition of this application had a significant antibacterial effect.

[0019] (3) Compared with single medicinal materials, the composition of this application has a stronger effect in inhibiting pro-inflammatory factors such as TNF-α, IL-1β, and IL-6;

[0020] (4) The most preferred formulation of the product of this application has free radical scavenging rates of 80.7% and 83.2% for DPPH and ABTS, respectively, indicating that the composition has high activity in scavenging free radicals;

[0021] (5) The sample in this application has excellent inhibitory effect on tyrosinase, that is, it has excellent whitening effect. Attached Figure Description

[0022] Figure 1 The results of the inhibition zone determination for Staphylococcus aureus in Example 1 are shown.

[0023] Figure 2The results of the inhibition zone assay for Propionibacterium acnes in Example 1 are shown.

[0024] Figure 3 The results of the inhibition zone determination for Trichophyton mentagrophytes in Example 1 are shown. Detailed Implementation

[0025] To enable those skilled in the art to better understand the present invention, the present invention will now be further described in conjunction with specific embodiments.

[0026] Example 1

[0027] This invention provides a traditional Chinese medicine composition with antibacterial, anti-inflammatory, antioxidant and whitening effects, which is prepared from Rhodiola rosea, licorice and Phyllanthus emblica in a mass ratio of 2:5:2.

[0028] The preparation method is as follows:

[0029] S1. Rhodiola rosea, licorice, and amla are pulverized and passed through a 20-mesh sieve to obtain powdered raw materials;

[0030] S2. Add the powdered raw material in S1 to caprylic / capric triglyceride at a mass-to-volume ratio of 1:10, extract at 110℃ for 1.0 h, filter through an 80-mesh filter cloth, discard the residue, collect the filtrate, and obtain the filtrate.

[0031] S3. Filter the filtrate obtained in S2 through a filter plate until the filtrate is clear. The filter plate has a pore size of 5 μm. The clear filtrate is the traditional Chinese medicine composition of the present invention.

[0032] Example 2

[0033] This invention provides a traditional Chinese medicine composition with antibacterial, anti-inflammatory, antioxidant and whitening effects, which is prepared from Rhodiola rosea, licorice and Phyllanthus emblica in a mass ratio of 5:10:5.

[0034] The preparation method is as follows:

[0035] S1. Rhodiola rosea, licorice, and amla are pulverized and passed through a 20-mesh sieve to obtain powdered raw materials;

[0036] S2. Add the powdered raw material from S1 to coconut oil alcohol-octanoate / decanoate at a mass-volume ratio of 1:2, extract at 180℃ for 0.5h, filter through a 100-mesh filter cloth, discard the residue, collect the filtrate, and obtain the filtrate.

[0037] S3. Filter the filtrate obtained in S2 through a filter plate until the filtrate is clear. The filter plate has a pore size of 10 μm. The clear filtrate is the traditional Chinese medicine composition of the present invention.

[0038] Example 3

[0039] This invention provides a traditional Chinese medicine composition with antibacterial, anti-inflammatory, antioxidant and whitening effects, which is prepared from Rhodiola rosea, licorice and Phyllanthus emblica in a mass ratio of 1:10:1.

[0040] The preparation method is as follows:

[0041] S1. Rhodiola rosea, licorice, and amla are pulverized and passed through a 10-mesh sieve to obtain powdered raw materials;

[0042] S2. Add the powdered raw material from S1 to olive oil at a mass-volume ratio of 1:20, extract at 50℃ for 3.0h, filter through an 80-mesh filter cloth, discard the residue, collect the filtrate, and obtain the filtrate.

[0043] S3. Filter the filtrate obtained in S2 through a filter plate until the filtrate is clear. The filter plate has a pore size of 2μm. The clear filtrate is the traditional Chinese medicine composition of the present invention.

[0044] Example 4

[0045] This invention provides a traditional Chinese medicine composition with antibacterial, anti-inflammatory, antioxidant and whitening effects, which is prepared from Rhodiola rosea, licorice and Phyllanthus emblica in a mass ratio of 1:9:4.

[0046] The preparation method is as follows:

[0047] S1. Rhodiola rosea, licorice, and amla are pulverized and passed through a 20-mesh sieve to obtain powdered raw materials;

[0048] S2. Add the powdered raw materials in S1 to phytosterol / octyldodecyl lauroyl glutamate at a mass-volume ratio of 1:8, extract at 100℃ for 1.5h, filter through a 100-mesh filter cloth, discard the residue, collect the filtrate, and obtain the filtrate.

[0049] S3. Filter the filtrate obtained in S2 through a filter plate until the filtrate is clear. The filter plate has a pore size of 8 μm. The clear filtrate is the traditional Chinese medicine composition of the present invention.

[0050] Example 5

[0051] This invention provides a traditional Chinese medicine composition with antibacterial, anti-inflammatory, antioxidant and whitening effects, which is prepared from Rhodiola rosea, licorice and Phyllanthus emblica in a mass ratio of 2:8:3.

[0052] The preparation method is as follows:

[0053] S1. Rhodiola rosea, licorice, and amla are pulverized and passed through a 10-mesh sieve to obtain powdered raw materials;

[0054] S2. Add the powdered raw material from S1 to camellia seed oil at a mass-volume ratio of 1:12, extract at 160℃ for 2.5 hours, filter through an 80-mesh filter cloth, discard the residue, collect the filtrate, and obtain the filtrate.

[0055] S3. Filter the filtrate obtained in S2 through a filter plate until the filtrate is clear. The filter plate has a pore size of 8 μm. The clear filtrate is the traditional Chinese medicine composition of the present invention.

[0056] Example 6

[0057] This invention provides a traditional Chinese medicine composition with antibacterial, anti-inflammatory, antioxidant and whitening effects, which is prepared from Rhodiola rosea, licorice and Phyllanthus emblica in a mass ratio of 2:6:3.

[0058] The preparation method is as follows:

[0059] S1. Rhodiola rosea, licorice, and amla are pulverized and passed through a 10-mesh sieve to obtain powdered raw materials;

[0060] S2. Add the powdered raw material from S1 to sweet almond oil at a mass-volume ratio of 1:16, extract at 150℃ for 1.5 hours, filter through a 100-mesh filter cloth, discard the residue, collect the filtrate, and obtain the filtrate.

[0061] S3. Filter the filtrate obtained in S2 through a filter plate until the filtrate is clear. The filter plate has a pore size of 10 μm. The clear filtrate is the traditional Chinese medicine composition of the present invention.

[0062] Comparative Example 1

[0063] This comparative example provides a product made from Rhodiola rosea alone. The amount of Rhodiola rosea used in Comparative Example 1 is the sum of the amounts of Rhodiola rosea, licorice, and Phyllanthus emblica used in Example 3. The preparation method is the same as in Example 3.

[0064] Comparative Example 2

[0065] This comparative example provides a product made from licorice alone. The amount of licorice used in Comparative Example 2 is the sum of the amounts of Rhodiola rosea, licorice, and Phyllanthus emblica used in Example 3, and its preparation method is the same as in Example 3.

[0066] Comparative Example 3

[0067] This comparative example provides a product made from Phyllanthus emblica alone. The amount of Phyllanthus emblica used in Comparative Example 3 is the sum of the amounts of Rhodiola rosea, licorice, and Phyllanthus emblica used in Example 3, and its preparation method is the same as in Example 3.

[0068] Comparative Example 4

[0069] This comparative example provides a product made from a traditional Chinese medicine composition consisting of Rhodiola rosea and licorice, prepared by Rhodiola rosea and licorice in a mass ratio of 1:10. The amount of Rhodiola rosea and licorice used in Comparative Example 4 is the sum of the amounts of Rhodiola rosea, licorice, and Phyllanthus emblica used in Example 3, and its preparation method is the same as in Example 3.

[0070] Comparative Example 5

[0071] This comparative example provides a product made from a traditional Chinese medicine composition consisting of licorice and amla, prepared by mixing licorice and amla in a mass ratio of 10:1. The amount of licorice and amla used in Comparative Example 5 is the sum of the amounts of rhodiola rosea, licorice, and amla used in Example 3, and its preparation method is the same as in Example 3.

[0072] Comparative Example 6

[0073] This comparative example provides a product made from a traditional Chinese medicine composition consisting of Rhodiola rosea and Phyllanthus emblica, prepared from Rhodiola rosea and Phyllanthus emblica in a mass ratio of 1:1. The amount of Rhodiola rosea and Phyllanthus emblica used in Comparative Example 6 is the sum of the amounts of Rhodiola rosea, Glycyrrhiza uralensis, and Phyllanthus emblica used in Example 3, and its preparation method is the same as in Example 3.

[0074] Comparative Example 7

[0075] This comparative example provides a product made from a traditional Chinese medicine composition consisting of Rhodiola rosea, licorice, and Phyllanthus emblica, prepared from Rhodiola rosea, licorice, and Phyllanthus emblica in a mass ratio of 1:10:1. The preparation method is basically the same as in Example 1, except that deionized water is used instead of caprylic / capric triglycerides during the extraction of the raw materials.

[0076] Comparative Example 8

[0077] This comparative example provides a product made from a traditional Chinese medicine composition consisting of Rhodiola rosea, licorice, and Phyllanthus emblica, prepared from Rhodiola rosea, licorice, and Phyllanthus emblica in a mass ratio of 1:10:1. The preparation method is basically the same as in Example 1, except that 70% ethanol is used instead of caprylic / capric triglycerides during the extraction of the raw materials.

[0078] Experimental results verification

[0079] Example 1

[0080] This example demonstrates the antibacterial effects of the traditional Chinese medicine compositions prepared in Examples 1-6 and Comparative Examples 1-8 of this invention.

[0081] 1. Experimental Methods

[0082] 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.

[0083] 2. Experimental Results

[0084] The results of the inhibition zone assay are shown in Table 1 and Figures 1-3 .

[0085] Table 1. Results of inhibition zone determination (n=3, )

[0086]

[0087]

[0088] Note: 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 Example 3, $ P<0.05, $$ P<0.01. Compared with Comparative Example 4, & P<0.05, && P < 0.01. Compared with comparative sample 5, Compared with comparative sample 6, § P<0.05, §§ P < 0.01. Compared with comparative sample 7, ￥ P<0.05, ￥￥ P<0.01. Compared with Comparative Example 8, α P<0.05, αα P<0.01.

[0089] According to the results in Table 1, compared with Comparative Examples 1-8, the compositions of Examples 1-6 showed significant antibacterial effects against Staphylococcus aureus, Propionibacterium acnes, and Trichophyton mentagrophytes (P<0.01), while Comparative Example 1 showed poor antibacterial effect and had no antibacterial effect against any of the three pathogens. Among them, the antibacterial effect of Example 3 was particularly outstanding, significantly better than the other examples and comparative examples. Each example showed the best inhibitory effect against Propionibacterium acnes. Furthermore, the examples showed significantly better effects than the comparative examples in terms of the diameter of the inhibition zone in the antibacterial experiment, both individually and in pairs. The specific ratio of Rhodiola rosea, licorice, and Phyllanthus emblica significantly improved the overall antibacterial efficacy of the traditional Chinese medicine composition, which was superior to the single application of each herb and their pairwise combinations. The experimental results show that the compatibility of the examples can effectively enhance the synergistic effect between the herbs, thus showing a significant advantage in antibacterial performance. In addition, the comparison results between Comparative Examples 7 and 8 and Example 3 show that although the formulation ratios of the three are the same, the difference in the extraction solvent leads to significantly different antibacterial effects. Oil solvents can better dissolve fat-soluble active ingredients in medicinal materials during extraction, such as certain flavonoids, which may play an important role in antibacterial mechanisms. Therefore, the oil solvent extraction system of this traditional Chinese medicine composition has potential application value as an antibacterial raw material in the field of skin care, especially suitable for inhibiting pathogens associated with skin infections.

[0090] Example 2

[0091] This example demonstrates the anti-inflammatory effects of the traditional Chinese medicine compositions prepared in Examples 1-6 and Comparative Examples 1-8 of this invention.

[0092] 1. Experimental Methods

[0093] 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. 5 RAW264.7 cells were seeded into 24-well plates (cells / well), with a blank control group, an LPS group, various drug-treated groups, and a positive control group (dexamethasone). After culturing for 12 h in a 5% CO2, 37°C incubator, the cells were pre-treated and protected for 12 h with samples from each embodiment of the present invention (20 μg / mL), comparative samples (20 μg / mL), and positive control (dexamethasone, 20 μg / mL). Then, RAW264.7 cells were induced with lipopolysaccharide (LPS, 1 μg / mL) for 24 h. After induction, the cell culture supernatant was collected, and the production of inflammatory factors was detected using an ELISA kit.

[0094] 2. Statistical methods

[0095] 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.

[0096] 3. Experimental Results

[0097] The results of the levels of pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 are shown in Table 2.

[0098] Table 2. Results of inflammatory markers (n=3, )

[0099]

[0100] Note: Compared with the blank group, Compared with the model group, 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 Example 3, $ P<0.05, $$ P<0.01. Compared with Comparative Example 4, & P<0.05, && P < 0.01. Compared with comparative sample 5, Compared with comparative sample 6, § P<0.05, §§ P < 0.01. Compared with comparative sample 7, ￥ P<0.05, ￥￥ P<0.01. Compared with Comparative Example 8, α P<0.05, αα P<0.01.

[0101] According to the results in Table 2, LPS stimulation significantly induced the expression of pro-inflammatory cytokines such as TNF-α, IL-1β, and IL-6 in RAW264.7 cells, and the levels of inflammatory factors in the model group were significantly increased (P<0.01), confirming that LPS successfully established an inflammation model. The traditional Chinese medicine compositions in Examples 1-6 all significantly inhibited the expression of these inflammatory factors (P<0.01), exhibiting significant anti-inflammatory effects. Example 3, in particular, showed the most outstanding performance in reducing TNF-α, IL-1β, and IL-6, which may be related to the unique ratio and synergistic effect of the medicinal materials.

[0102] Compared with Comparative Examples 1-8, the Example group showed a more significant effect in inhibiting inflammatory factors (P<0.01), indicating that the ratio and interaction of the medicinal materials in this invention greatly enhance the anti-inflammatory activity of the composition, which is superior to that of a single medicinal material or a combination of two medicinal materials. In particular, the direct comparison of Comparative Example 7 (water extraction), Comparative Example 8 (70% ethanol extraction), and Example 3 (oil solvent extraction) further highlights the influence of the extraction solvent on the anti-inflammatory properties. Although Comparative Examples 7, 8, and Example 3 have the same formulation, the effects of Comparative Examples 7 and 8 in inhibiting pro-inflammatory factors such as TNF-α, IL-1β, and IL-6 are significantly weaker than those in Example 3 because Comparative Examples 7 and 8 used water and 70% ethanol as extraction solvents. These results provide strong scientific evidence for the development of highly effective anti-inflammatory skin products and have broad application potential.

[0103] Example 3

[0104] This example demonstrates the antioxidant effects of the traditional Chinese medicine compositions prepared in Examples 1-6 and Comparative Examples 1-8 of the present invention.

[0105] 1. Experimental Methods

[0106] 1.1 Effect on DPPH free radical scavenging

[0107] Taking the sample prepared in Example 1 as an example, its DPPH radical scavenging rate was tested as follows:

[0108] The control group was set up as follows: 3 mL of DPPH solution was thoroughly mixed with 1 mL of pure water to obtain a mixed solution, and left to stand for 30 min. Then, 200 μL of the mixed solution was transferred into a 96-well plate using a pipette, and the results were repeated for 3 replicates. The absorbance at 517 nm was then measured using an ELISA reader, and the average value was taken as A1.

[0109] The experimental setup was as follows: 3 mL of DPPH solution was thoroughly mixed with 1 mL of the sample prepared in Example 1 to obtain a mixed solution. After standing for 30 min, 200 μL of the mixed solution was transferred into a 96-well plate using a pipette. This was repeated for 3 replicates. The absorbance at 517 nm was then measured using an ELISA reader, and the average value was taken as A2.

[0110] The blank group is set as follows: no sample is placed in the 96-well plate, and the absorbance at 517 nm is directly measured.

[0111] DPPH removal rate = ((A1-A0)-(A2-A0)) / (A1-A0)×100%.

[0112] The samples prepared in Example 1 were replaced with the samples corresponding to Examples 2-8 and Comparative Examples 1 and 2, respectively. The absorbance values ​​were measured and the DPPH free radical scavenging rate was calculated.

[0113] 1.2 ABTS + Effects of free radical scavenging

[0114] 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 in 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.

[0115] The calculation formula is as follows:

[0116]

[0117] A0: Replace the sample with the same volume of distilled water, and follow the same procedure as A1.

[0118] 2. Experimental Results

[0119] 2.1 Analysis of in vitro antioxidant assay results

[0120] Table 3 Results of in vitro antioxidant activity determination

[0121]

[0122] The results in Table 3 show that the traditional Chinese medicine compositions obtained in Examples 1-6 all exhibited excellent DPPH and ABTS free radical scavenging abilities, clearly indicating that these compositions possess significant antioxidant activity. Overall, the antioxidant effects of the examples were generally superior to those of Comparative Examples 1-8, further verifying the significant advantages of these traditional Chinese medicine compositions in antioxidant properties.

[0123] Specifically, Example 3 performed particularly well, achieving DPPH and ABTS free radical scavenging rates of 80.7% and 83.2%, respectively, indicating that the composition exhibits high activity in scavenging free radicals. This may be closely related to the synergistic effect brought about by its specific herbal ratio, allowing the antioxidant components to exert their full effect.

[0124] In contrast, the antioxidant effects of Comparative Examples 1-8 were relatively weak, with DPPH and ABTS scavenging rates all below 60%, indicating that these compositions were less active in antioxidant activity than the examples. This may be due to insufficient optimization of the herbal formulation, failing to fully utilize the potential of the antioxidant components. In particular, the comparison between Comparative Examples 7 (using water extraction), 8 (using 70% ethanol extraction), and Example 3 (using oil solvent extraction) further highlights the influence of the extraction solvent on antioxidant performance. Although the herbal formulations of Comparative Examples 7, 8, and 3 were identical, the DPPH and ABTS scavenging abilities of Comparative Examples 7 and 8 were significantly weaker than those of Example 3.

[0125] In summary, the traditional Chinese medicine compositions of Examples 1 to 6 exhibited significant antioxidant capacity, especially Examples 2 and 3, which showed the best performance in DPPH and ABTS free radical scavenging experiments.

[0126] Example of effect 4

[0127] This example provides the whitening effect (evaluation of tyrosinase inhibition effect) of the traditional Chinese medicine compositions prepared in Examples 1-6 and Comparative Examples 1-8 of the present invention.

[0128] 1. Experimental Methods

[0129] (1) Preparation of phosphate buffer: Accurately weigh 26.797 g of Na2HPO4·12H2O and 13.803 g of NaH2PO4·2H2O, dissolve them separately in deionized water, and then dilute to 500 mL in a volumetric flask. Add 100 mL each of sodium dihydrogen phosphate and disodium hydrogen phosphate solution to a 500 mL volumetric flask, dilute to volume with deionized water, and shake well. This is the phosphate buffer solution with pH = 6.8.

[0130] (2) Preparation of sample solutions: The samples of each example and comparative example were prepared into test sample solutions of corresponding concentrations using DMSO.

[0131] (3) Preparation of substrate L-tyrosine solution: The substrate L-tyrosine was prepared into a 0.05% L-tyrosine solution using phosphate buffer.

[0132] (4) Preparation of tyrosinase solution: Tyrosinase was prepared into a 100 U / mL solution using phosphate buffer.

[0133] (5) Determination of tyrosinase inhibition rate: Accurately pipette the four reaction sample solutions (C1, C2, T1, and T2) excluding the tyrosinase solution according to Table 4. After incubating at 37℃ for 10 minutes, add the tyrosinase solution and react for another 10 minutes. Using C2 as the reference for C1, measure the absorbance A1 of the system at 475 nm. Using T2 as the reference for T1, measure the absorbance A2 of the system. To compensate for systematic errors caused by parallel samples, 1 mL of DMSO was added to C1 and C2 respectively. To eliminate the absorbance error 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 results for each sample.

[0134] The results are shown in Table 5.

[0135]

[0136] 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.

[0137] Table 4. Reagent dosage for the tyrosinase activity inhibition experiment.

[0138]

[0139] Table 5. Results of tyrosinase activity inhibition rate (IC50, n=3). )

[0140]

[0141] Note: 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 Example 3, $ P<0.05, $$ P<0.01. Compared with Comparative Example 4, & P<0.05, && P < 0.01. Compared with comparative sample 5, Compared with comparative sample 6, § P<0.05, §§ P < 0.01. Compared with comparative sample 7, ￥ P<0.05, ￥￥ P<0.01. Compared with Comparative Example 8, α P<0.05, αα P<0.01.

[0142] As shown in Table 5, the samples from Examples 1-6 exhibited significantly better inhibitory effects on tyrosinase than the comparative samples 1-8 (P<0.01). Compared to the comparative samples, the example samples demonstrated stronger tyrosinase inhibitory activity, as evidenced by their lower IC50 values. This indicates that the example samples possess superior tyrosinase inhibitory effects. Therefore, the examples can be added as cosmetic ingredients to effectively enhance the whitening efficacy of the final product.

[0143] The samples in the example group showed a more significant inhibitory effect on tyrosinase activity, indicating that the raw material ratio and interaction in this invention greatly enhance the whitening activity of the composition, which is superior to that of a single herb or a combination of two herbs. In particular, the direct comparison between Comparative Example 7 (water extraction), Comparative Example 8 (70% ethanol extraction), and Example 3 (oil solvent extraction) further highlights the whitening effect of the combined oil-soluble components.

Claims

1. A traditional Chinese medicine composition that simultaneously possesses antibacterial, anti-inflammatory, antioxidant, and whitening effects, prepared from Rhodiola rosea, licorice, and Phyllanthus emblica in a mass ratio of 1:10:1; The preparation method of the traditional Chinese medicine composition is as follows: S1. Rhodiola rosea, licorice, and amla are pulverized and passed through a 10-mesh sieve to obtain powdered raw materials; S2. Add the powdered raw material from S1 to olive oil at a mass-volume ratio of 1 g: 20 mL, extract at 50℃ for 3.0 h, filter through an 80-mesh filter cloth, discard the residue, collect the filtrate, and obtain the filtrate. S3. Filter the filtrate obtained in S2 until it becomes clear. The filter plate used for filtration has a pore size of 2μm. The resulting clear filtrate is the traditional Chinese medicine composition.

2. The application of the traditional Chinese medicine composition as described in claim 1 in the preparation of antibacterial products.

3. The use of the traditional Chinese medicine composition as described in claim 1 in the preparation of anti-inflammatory products.

4. The use of the traditional Chinese medicine composition as described in claim 1 in the preparation of antioxidant products.

5. The application of the traditional Chinese medicine composition as described in claim 1 in the preparation of skin whitening products.

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