A traditional Chinese medicine composition with acne-removing and soothing effects, a preparation method and application thereof

Abstract

This invention relates to the field of cosmetic technology, specifically disclosing a traditional Chinese medicine composition with acne-removing and soothing effects, its preparation method, and its application. The traditional Chinese medicine composition is a compound of extracts of Myrtle, Polygonum cuspidatum, Salvia miltiorrhiza, and Dictamnus dasycarpus in a mass ratio of (2-4):(2-4):(1-2):(1-2). This traditional Chinese medicine composition can significantly inhibit the growth of Propionibacterium acnes, Staphylococcus aureus, and Staphylococcus epidermidis, inhibit sebum secretion, and significantly inhibit hyaluronidase activity. It achieves acne-removing effects through a synergistic effect of antibacterial, oil-controlling, anti-inflammatory, and soothing properties. Skin irritation tests show that this traditional Chinese medicine composition is highly safe and non-irritating. This traditional Chinese medicine composition can be used to prepare topical products with acne-removing, oil-controlling, and soothing effects, including cosmetic dosage forms such as gels, creams, serums, and masks, as well as topical pharmaceutical preparations such as ointments and lotions, and has good market application prospects.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a traditional Chinese medicine composition with acne-removing and soothing effects, its preparation method, and its application. Background Technology

[0002] Acne vulgaris (commonly known as pimples) is a common chronic inflammatory skin disease with a global incidence rate as high as 85% among adolescents and a persistently high prevalence among adults. This disease not only manifests as skin lesions such as blackheads, papules, and pustules, but can also leave permanent scars, severely impacting patients' quality of life and mental health. Its harm extends far beyond the cosmetic appearance; it is often accompanied by psychological problems such as anxiety, social phobia, and decreased self-confidence. Related clinical studies show that the negative impact of acne on patients' emotions and quality of life can, in severe cases, be comparable to that of chronic systemic diseases such as diabetes and epilepsy.

[0003] The pathogenesis of acne involves the interaction of multiple factors, including microbial infection, excessive sebum secretion, abnormal follicular keratinization, and local inflammatory response. Among these, Propionibacterium acnes is considered a core pathogenic factor. The abnormal colonization and proliferation of this bacterium within the hair follicle can trigger a local inflammatory cascade, promoting the development of comedones into inflammatory papules and pustules.

[0004] Excessive sebum secretion is closely related to the aforementioned infection process: excess sebum provides a rich nutrient substrate for lipophilic Propionibacterium acnes, promoting its proliferation; simultaneously, the lipases secreted by this bacterium can break down triglycerides in sebum, releasing free fatty acids. These free fatty acids not only stimulate excessive keratinization of the hair follicle opening, worsening blockage, but can also trigger a strong inflammatory response, thus forming a vicious cycle of "sebum overflow - bacterial proliferation - increased inflammation." Therefore, simultaneously intervening in the two key aspects of "antibacterial control" and "oil control," and breaking this vicious cycle at its source, is an important strategy for the scientific prevention and treatment of acne.

[0005] Currently, clinical treatment for acne mainly relies on two approaches: chemical drugs and traditional Chinese medicine. In chemical drug treatment, while antibiotics (such as clindamycin) and retinoids can inhibit bacterial proliferation or regulate keratin metabolism, long-term use can easily lead to drug resistance, skin barrier damage, and systemic side effects. For example, the clinical resistance rate to antibiotics is on the rise, while retinoids may cause dry skin and even pose a teratogenic risk, limiting their widespread use. Furthermore, while some chemical exfoliating products (such as salicylic acid) can quickly improve skin lesions, they may damage the integrity of the epidermis, exacerbating skin sensitivity and moisture loss, failing to meet current demands for "safe, gentle, and long-lasting" skincare.

[0006] In recent years, traditional Chinese medicine compound formulas have attracted much attention in the development of acne treatment products due to their advantages such as multi-target effects, low irritation, and natural safety. However, existing technologies still have the following limitations: First, the formulation design lacks systematicity and synergy. Most existing products or patents simply combine a few "heat-clearing and detoxifying" herbs, resulting in a piling-up effect with unclear primary and secondary components. They fail to scientifically screen and optimize based on clear antibacterial spectra, oil-control mechanisms, or anti-inflammatory pathways. For example, some compound formulas claim broad-spectrum antibacterial activity, but in reality, their inhibitory effect on the core acne pathogen—Propionibacterium acnes—is weak; or they only focus on antibacterial activity while neglecting the synergistic effects of oil control and barrier repair, leading to short-lasting efficacy and easy recurrence.

[0007] Secondly, the compatibility between extraction processes and dosage forms is insufficient. Traditional Chinese medicine research is often constrained by the mindset of oral administration, with extraction processes long limited to water or ethanol as solvents, primarily due to considerations of oral bioavailability. However, when developing topical formulations, the transdermal absorption patterns of the skin barrier, the local retention of active ingredients, and compatibility with cosmetic matrices place drastically different demands on extraction solvents, leading to insufficient compatibility between traditional processes and topical dosage forms. Research on stepwise solvent extraction adapted to the characteristics of topical dosage forms remains extremely limited.

[0008] Furthermore, although ultrasonic extraction technology is now widespread, its high efficiency, low temperature, and time-saving characteristics offer a new approach for the gentle extraction of active ingredients. However, there are few reports on existing technologies that combine ultrasonic extraction with adaptable stepwise solvent extraction methods and systematically optimize the extraction process for external use.

[0009] Meanwhile, traditional water or alcohol extraction processes often require complex concentration steps after extraction to remove large amounts of solvent, which not only increases energy consumption and cost but may also deactivate heat-sensitive components. Therefore, developing an adaptable stepwise solvent extraction process that integrates the advantages of multi-solvent screening and ultrasonic extraction while avoiding cumbersome post-processing steps is a key technological bridge connecting the activity of traditional Chinese medicine with the efficacy of cosmetics.

[0010] Furthermore, their efficacy coverage is relatively limited. Most existing products focus only on single aspects such as antibacterial or oil control, neglecting the complex characteristics of the acne pathogenesis, where microbial colonization, sebum secretion, and inflammatory response mutually reinforce each other. Intervention targeting a single point often has limited effects and cannot meet the clinical needs for synergistic prevention and treatment across multiple stages.

[0011] In summary, existing chemical acne treatments suffer from significant side effects and are prone to drug resistance, while traditional Chinese medicine (TCM) acne treatment technologies face shortcomings such as unscientific formulation, poor compatibility between extraction processes and topical dosage forms, and limited efficacy coverage. Neither approach can achieve safe, efficient, and comprehensive prevention and treatment of acne. Therefore, developing a TCM composition with natural raw materials, a rational formulation, advanced technology, and the ability to simultaneously intervene in multiple pathological stages of acne has become a key direction for overcoming existing technological bottlenecks and meeting the core clinical and market demands for acne treatment products. Summary of the Invention

[0012] The purpose of this invention is to overcome the limitations and shortcomings of existing products that mostly focus on single effects such as antibacterial or oil control, and to provide a scientifically formulated and synergistically effective traditional Chinese medicine composition with acne-removing and soothing effects. The composition has multiple effects, including broad-spectrum antibacterial, effective regulation of sebum secretion, and significant anti-inflammatory and soothing effects. It can simultaneously intervene in the three key links of acne occurrence and development: "microbial infection", "excessive sebum secretion" and "inflammatory response", thereby meeting the clinical needs of comprehensive acne prevention and treatment.

[0013] The second objective of this invention is to provide a method for preparing the traditional Chinese medicine composition with acne-removing and soothing effects.

[0014] A third objective of this invention is to provide the application of the traditional Chinese medicine composition having acne-removing and soothing effects.

[0015] The above-mentioned objective of this invention is achieved through the following technical solution: A traditional Chinese medicine composition with acne-removing and soothing effects is characterized in that the composition is composed of extracts of Myrtle, Polygonum cuspidatum, Salvia miltiorrhiza, and Dictamnus dasycarpus in a mass ratio of (2-4):(2-4):(1-2):(1-2); the Myrtle, Polygonum cuspidatum, and Salvia miltiorrhiza extracts are plant oil extracts obtained by using cosmetic-grade plant oils as extraction solvents; and the Dictamnus dasycarpus extract is a propylene glycol extract obtained by using propylene glycol as an extraction solvent.

[0016] Cosmetic-grade plant oils are oil-phase raw materials mainly composed of triglycerides extracted from plant seeds, fruits (and a few from leaves / barks / roots / flowers), which are refined / decolorized / deodorized to meet the purity, stability and safety requirements of cosmetics.

[0017] This invention takes into account the widely used plant oils (such as sweet almond oil and jojoba oil) and polyols (such as propylene glycol and butylene glycol) in the cosmetics industry. These not only possess excellent skin compatibility, moisturizing, and penetration-enhancing effects, but also exhibit superior extraction efficiency for lipid-soluble or specific polar active ingredients in traditional Chinese medicine. For example, using oils as solvents can efficiently dissolve lipid-soluble components such as tanshinone and phloroglucinol. More importantly, the extract obtained after extraction with such solvents has excellent compatibility with common cosmetic matrices (oil phase, aqueous phase, or emulsion system), and can be directly used in formulations without the need for complex "extraction-concentration-redissolution" steps. This significantly reduces energy consumption and minimizes damage to heat-sensitive components. However, existing technologies rarely systematically study and apply such cosmetic raw materials as extraction solvents for traditional Chinese medicine, and their potential as extraction solvents has not been fully explored.

[0018] To address the shortcomings of existing technologies, this invention systematically screened the in vitro activity of 29 kinds of traditional Chinese medicinal herbs (such as Myrtle, Polygonum cuspidatum, and Salvia miltiorrhiza). It discovered that a traditional Chinese medicine composition consisting of Myrtle oil extract, Polygonum cuspidatum oil extract, Salvia miltiorrhiza oil extract, and Dictamnus dasycarpus root bark propylene glycol extract in a specific ratio exhibits significant acne-removing effects. It can effectively inhibit Propionibacterium acnes and related pathogens (Staphylococcus aureus and Staphylococcus epidermidis). Simultaneously, it possesses oil-controlling (inhibiting sebum secretion) and soothing (inhibiting hyaluronidase activity) effects, with each component demonstrating a clear synergistic effect. This achieves simultaneous intervention in the three key stages of acne pathogenesis: "microbial infection," "excessive sebum secretion," and "inflammatory response." In other words, it achieves acne removal through a triple effect of "antibacterial, oil-controlling, and anti-inflammatory soothing," overcoming the problem of single-effect products in existing products.

[0019] Among them, myrtle ( Rhodomyrtus tomentosa (Ait.) Hassk. is a plant belonging to the genus *Myrtle* in the family Myrtaceae. Its leaves, fruits, and other parts are rich in active ingredients such as phloroglucinols, flavonoids, tannins, and polyphenols. Modern pharmacological studies have shown that *Myrtle* extract possesses significant antibacterial, anti-inflammatory, and antioxidant activities. Among these, the characteristic component rhodomyrtone and various phloroglucinol derivatives exhibit potent inhibitory effects against Gram-positive bacteria and *Propionibacterium acnes*, with mechanisms including disruption of bacterial biofilm formation and interference with cell function. Furthermore, gallic acid, flavonoid glycosides, and anthocyanins in *Myrtle* fruit extract effectively scavenge free radicals, inhibit sebum oxidation, reduce pathological damage to skin cells, and also possess anti-inflammatory and soothing effects.

[0020] Salvia miltiorrhiza ( Salvia miltiorrhizaSalvia miltiorrhiza (Bge.) is the dried root and rhizome of a plant in the Lamiaceae family, genus Salvia, and is a traditional Chinese medicine for promoting blood circulation and removing blood stasis. The main active components of Salvia miltiorrhiza are divided into lipid-soluble tanshinones (such as cryptotanshinone and tanshinone IIA) and water-soluble phenolic acids (tanshinone acid A, tanshinone acid B, and tanshinone). Modern pharmacological studies have shown that tanshinone components are highly sensitive to Propionibacterium acnes, and their antibacterial activity is related to inhibiting bacterial biofilm formation and interfering with cell metabolism. Simultaneously, tanshinone has anti-androgenic effects, effectively downregulating the sebaceous gland's responsiveness to androgens, inhibiting excessive sebaceous gland proliferation and lipid secretion, thereby reducing sebum secretion at its source. Furthermore, tanshinone and tanshinone acid components significantly reduce the release of pro-inflammatory factors such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and prostaglandin E2 (PGE2) by regulating the NF-κB, MAPK, and NLRP3 inflammasome signaling pathways, exhibiting non-steroidal anti-inflammatory activity similar to hydrocortisone. The components of salvianolic acid also have the effects of effectively scavenging free radicals, inhibiting lipid peroxidation and promoting fibroblast repair, which helps to reduce post-acne erythema and pigmentation and promote barrier repair.

[0021] Polygonum cuspidatum ( Polygonum cuspidatum *Polygonum cuspidatum* (Sieb. et Zucc.) is the dried rhizome and root of a plant in the Polygonaceae family. It is slightly bitter and slightly cold in nature, and enters the liver, gallbladder, and lung meridians. Traditionally, it has the effects of clearing heat and detoxifying, promoting diuresis and relieving jaundice, and dispersing blood stasis and relieving pain. Modern research shows that *Polygonum cuspidatum* is rich in anthraquinones (such as emodin and emodin methyl ether), stilbenesides (such as resveratrol and resveratrol glycoside), and flavonoids, among other active ingredients. These components collectively endow it with significant antibacterial, anti-inflammatory, and antioxidant effects. In terms of antibacterial activity, *Polygonum cuspidatum* inhibits various skin pathogens such as *Staphylococcus aureus* and *Propionibacterium acnes*. Its anti-inflammatory mechanism involves inhibiting signaling pathways such as NF-κB and reducing the expression of pro-inflammatory factors such as TNF-α and IL-6. Simultaneously, *Polygonum cuspidatum* extract can effectively scavenge free radicals and reduce oxidative stress damage.

[0022] Dictamnus dasycarpus is the root bark of the Rutaceae plant Dictamnus dasy Dictamnus dasycarpus The dried root bark of *Dictamnus dasycarpus* is bitter and cold in nature, and enters the spleen, stomach, and bladder meridians. It has the traditional effects of clearing heat and drying dampness, dispelling wind and detoxifying. Modern research shows that *Dictamnus dasycarpus* root bark has a comprehensive effect on multiple targets and pathways in the treatment of skin-related diseases, and is particularly suitable for the prevention and treatment of acne. First, *Dictamnus dasycarpus* root bark is rich in various alkaloids such as dictamnus alkaloids, francone, and berberine, as well as limonene compounds, which show significant in vitro antibacterial activity against various skin pathogens and dermatophytes that cause folliculitis, effectively inhibiting the proliferation of microorganisms such as *Propionibacterium acnes*. Second, its extract can significantly inhibit various non-specific inflammations and reduce the release of pro-inflammatory factors such as TNF-α and IL-6 by downregulating signaling pathways such as NF-κB and MAPK, thereby exerting anti-inflammatory and soothing effects and relieving redness and pain caused by acne.

[0023] The extraction solvents used in this invention are all selected from commonly used basic raw materials in the cosmetics field. Among them, plant oils (such as sweet almond oil) have good solubility and skin affinity for the fat-soluble active ingredients (such as tanshinone, phloroglucinol, and resveratrol) in myrtle, knotweed, and salvia miltiorrhiza; propylene glycol has a high extraction efficiency for moderately polar active ingredients such as alkaloids and limonenes in dictamnus dasycarpus. Because the selected solvents are all commonly used raw materials in cosmetics, the resulting extract has good compatibility with cosmetic matrices and can be directly applied to the formulation of cosmetic dosage forms without concentration, significantly simplifying the process and reducing energy consumption.

[0024] Furthermore, the cosmetic-grade plant oil is selected from one or more of the following: sweet almond oil, peach kernel oil, olive oil, grape seed oil, jojoba oil, safflower seed oil, sunflower seed oil, avocado oil, and meadowfoam seed oil. Preferably, the cosmetic-grade plant oil is sweet almond oil, grape seed oil, or jojoba oil.

[0025] The present invention also provides a method for preparing the aforementioned traditional Chinese medicine composition, comprising the following steps: S1. Using cosmetic-grade plant oils as extraction solvents, extract the three medicinal herbs Myrtle, Polygonum cuspidatum, and Salvia miltiorrhiza, and obtain the corresponding plant oil extracts after centrifugation. S2. Using propylene glycol as the extraction solvent, extract the medicinal material of Dictamnus dasycarpus, and obtain the propylene glycol extract of Dictamnus dasycarpus after centrifugation and filtration. S3. Mix the extracts of Myrtle, Polygonum cuspidatum, Salvia miltiorrhiza and Dictamnus dasycarpus obtained in the above steps at a mass ratio of (2-4):(2-4):(1-2):(1-2) and disperse them to obtain the Chinese herbal composition.

[0026] Furthermore, in steps S1 and S2, the ratio of medicinal material to extraction solvent is 1:10-15 (w / v).

[0027] Furthermore, the extraction described in steps S1 and S2 is ultrasonic extraction, with an extraction temperature of 25–50°C, an ultrasonic power of 400–500 W, and an extraction time of 60–90 minutes.

[0028] As a preferred embodiment, the preparation method of the traditional Chinese medicine composition specifically includes the following steps: (1) Preparation method of myrtle extract: It is prepared by ultrasonic-assisted extraction process. Cosmetic grade plant oil is used as solvent. Myrtle powder is added at a material-liquid ratio of 1:10-15 (medicinal material weight g: solvent volume mL, w / v). It is extracted for 60-90 minutes at ultrasonic power of 400-500W and temperature of 25-50℃. After removing the residue by coarse filtration, the extract is centrifuged at a speed of 15000-20000r / min using a tubular centrifuge. The supernatant obtained is myrtle extract.

[0029] (2) Preparation method of Polygonum cuspidatum extract: Same as myrtle extract.

[0030] (3) Preparation method of Danshen extract: Same as myrtle extract.

[0031] (4) Preparation method of Dictamnus dasycarpus root bark extract: The extract was prepared using an ultrasonic-assisted extraction process. Propylene glycol was used as the solvent, and crude powder of Dictamnus dasycarpus root bark was added at a material-to-liquid ratio of 1:10-15 (herb weight g: solvent volume mL, w / v). The extract was extracted for 60-90 minutes at an ultrasonic power of 400-500 W and a temperature of 25-50℃. After coarse filtration to remove the residue, the extract was centrifuged at 15000-20000 r / min using a tubular centrifuge. The supernatant obtained was the Dictamnus dasycarpus root bark extract.

[0032] (5) Preparation of the composition: The extracts of Myrtle, Polygonum cuspidatum, Salvia miltiorrhiza and Dictamnus dasycarpus obtained in the above steps are mixed in a mass ratio of (2-4):(2-4):(1-2):(1-2) and dispersed by mechanical stirring to obtain the Chinese herbal composition.

[0033] The preparation method aims to address the shortcomings of traditional Chinese medicine extraction processes (such as water extraction and alcohol extraction) that are incompatible with the needs of cosmetics and topical preparations. By employing a "compatibility-based stepwise solvent extraction" process, cosmetic-grade plant oils (such as sweet almond oil) and propylene glycol are selected as extraction solvents based on the polarity differences of active ingredients in different medicinal materials, combined with ultrasound-assisted technology. This process not only efficiently extracts the target active ingredients, but also produces an extract with excellent compatibility with cosmetic matrices. It eliminates the need for complex concentration and solvent replacement steps, allowing direct application in dosage form formulation, thereby simplifying the production process, reducing energy consumption, and better preserving the activity of the ingredients.

[0034] The adaptive stepwise solvent ultrasonic extraction process efficiently extracts the target components under mild conditions (25-50℃). The process is simple, has high extraction efficiency and low energy consumption, avoids the damage of heat-sensitive components to high temperatures, and the obtained extract can be directly used for cosmetic formulation without concentration, making it suitable for industrial production.

[0035] The above-mentioned traditional Chinese medicine composition of this invention can significantly inhibit the growth of Propionibacterium acnes, Staphylococcus aureus, and Staphylococcus epidermidis, suppress sebum secretion, and significantly inhibit hyaluronidase activity. It achieves a comprehensive acne-removing effect through the synergistic effects of antibacterial, oil-controlling, anti-inflammatory, and soothing properties. Skin irritation tests show that this composition is highly safe and non-irritating.

[0036] Although the composition obtained by this invention is an oil-polar phase mixture, its solvents are all commonly used cosmetic raw materials, and with the assistance of solubilization / emulsification technology, it can be easily integrated into various topical dosage forms. The traditional Chinese medicine composition described in this invention can be used to prepare topical products with acne-removing, oil-controlling, antibacterial, and soothing effects. This composition can be applied to the preparation of skincare products or topical pharmaceutical preparations. Skincare products include acne-removing gels, acne-removing serums, acne-removing masks, acne-removing creams, etc.; topical pharmaceutical preparations include acne-removing ointments, acne-removing lotions, etc. This provides effective raw materials for the development of safe, efficient, and multi-dosage-form acne-removing products.

[0037] Therefore, the present invention also provides the use of any of the above-described traditional Chinese medicine compositions in the preparation of topical products for acne treatment, oil control, antibacterial and / or soothing.

[0038] The present invention also provides a topical product with acne-removing, oil-controlling, antibacterial and / or soothing effects, wherein the topical product contains any of the above-described traditional Chinese medicine compositions.

[0039] Furthermore, the mass content of the traditional Chinese medicine composition is 2-4%.

[0040] Preferably, the mass content of the traditional Chinese medicine composition is 3%.

[0041] Furthermore, the external products include, but are not limited to, cosmetic formulations such as gels, serums, creams, masks, and toners, as well as external pharmaceutical preparations such as ointments and lotions.

[0042] Compared with the prior art, the present invention has the following beneficial effects. 1. It achieves synergistic intervention targeting multiple pathological stages of acne, with a more comprehensive mechanism of action. The traditional Chinese medicine composition of this invention, through scientific formulation, achieves simultaneous intervention in three key aspects of acne pathogenesis: "microbial infection," "excessive sebum secretion," and "inflammatory response." Research shows that this composition not only has a significant inhibitory effect on the core acne pathogen—Propionibacterium acnes—and its related pathogens (Staphylococcus aureus, Staphylococcus epidermidis) (inhibition zone diameter reaches 17.9–18.8 mm), but also effectively inhibits sebum secretion (reduction rate reaches 17.7%–18.3%) and significantly inhibits hyaluronidase activity (inhibition rate 61.57%–65.48%). In other words, the traditional Chinese medicine composition can achieve acne removal through a triple effect of "antibacterial, oil control, and anti-inflammatory soothing," overcoming the problem of single efficacy in existing products.

[0043] 2. The formulation has undergone systematic screening and optimization, demonstrating a clear synergistic effect. The formulation of the traditional Chinese medicine composition of this invention is not a simple combination of medicinal materials. Through systematic screening of 29 kinds of traditional Chinese medicinal materials and their different solvent extracts, it was found that the combination of myrtle oil extract, Polygonum cuspidatum oil extract, Salvia miltiorrhiza oil extract and Dictamnus dasycarpus propylene glycol extract in a specific mass ratio of (2-4):(2-4):(1-2):(1-2) showed a significantly better antibacterial effect against Propionibacterium acnes than the individual extracts and other arbitrary combinations. This indicates that the combination of these four medicinal materials produced a clear synergistic effect of "1+1+1+1>4", rather than a simple linear additive effect.

[0044] 3. Innovative extraction process, combining high-efficiency extraction, dosage form compatibility, and ease of production. This invention breaks through the conventional thinking of traditional Chinese medicine extraction, which primarily uses water or ethanol, and creatively develops a "compatibility-adaptive stepwise solvent extraction process." The core innovation of this process lies in the use of cosmetic-grade plant oils (for extracting fat-soluble components from *Myrtle*, *Polygonum cuspidatum*, and *Salvia miltiorrhiza*) and propylene glycol (for extracting active components from *Dictamnus dasycarpus*) for ultrasonic extraction, based on the polarity differences of the active ingredients in the medicinal materials. This design offers three advantages: First, it improves the extraction efficiency of the target active ingredients; second, the selected solvents themselves have good skin affinity and permeation-enhancing properties, and excellent compatibility with common cosmetic bases (oil, water, emulsions), allowing the obtained extract to be directly used in the formulation of end products without complex concentration and solvent replacement steps, simplifying the process, reducing energy consumption, and avoiding damage to heat-sensitive components during post-processing; third, the process conditions are mild (25–50°C), and combined with ultrasonic assistance, it is highly efficient and suitable for large-scale production.

[0045] 4. It has good safety and is suitable for a wide range of people. According to the skin irritation test under the "Cosmetic Safety Technical Specifications" standard, the herbal composition of this invention is non-irritating to the skin (average irritation score of 0), and all ingredients are natural Chinese medicinal materials without chemical additives, avoiding the risk of drug resistance and side effects, and is especially suitable for people with sensitive skin and those who need long-term use.

[0046] 5. It has broad application prospects and significant market value. The herbal composition of this invention can be used as a core active ingredient in various skincare products (such as acne gels and creams) and topical pharmaceutical preparations (such as ointments and lotions), resulting in a variety of product forms. Its characteristics of "natural origin, multi-functionality, safety, and gentleness" highly align with the current market trend towards "efficacy-driven skincare" and "green ingredients," giving it significant commercial potential and market competitiveness. Attached Figure Description

[0047] Figure 1 This is a graph showing the inhibitory effect of Example 1 on Staphylococcus aureus. The numbers 1 through 4 represent four replicates.

[0048] Figure 2 This is a graph showing the inhibitory effect of Example 1 on Staphylococcus epidermidis. The numbers 1 through 4 represent four replicates.

[0049] Figure 3 This is a graph showing the inhibitory effect of Example 1 on Propionibacterium acnes. The numbers 1 through 4 represent four replicates. Detailed Implementation

[0050] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0051] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0052] Sources of medicinal materials and reagents: The four test medicinal materials in the examples and comparative examples were all commercially available Chinese medicinal decoction pieces, purchased from Guangzhou Xiwotang Trading Co., Ltd.

[0053] The sweet almond oil was purchased from GUSTAV HEESS GmbH, product number: 006655-00601-3975.

[0054] Propylene glycol was purchased from SK picglobal Co., Ltd., product number: 001383-00420-7144.

[0055] Carbomer 940 was purchased from Lubrizol Advanced Materials Co., Ltd., part number: 004079-00873-8683.

[0056] Glycerin was purchased from KL-KEPONG OLEOMAS SDN BHD, product number: 002421-03185-5838.

[0057] The PEG-40 hydrogenated castor oil was purchased from BASF (China) Co., Ltd., product number: 000681-01445-7410.

[0058] Triethanolamine was purchased from Dow Chemical (Shanghai) Co., Ltd., product number: 005819-00776-5408.

[0059] p-Hydroxyacetophenone was purchased from Nanjing Huashi New Materials Co., Ltd., item number: 002021-00751-6651.

[0060] 1,2-Hexanediol was purchased from B&B CO.,LTD, catalog number: 000004-01417-8148.

[0061] 1,3-Butanediol was purchased from Daicel Corporation, catalog number: 001946-02238-5448.

[0062] Sodium hyaluronate was purchased from Shandong Focus Freda Biotechnology Co., Ltd., product number: 006722-00607-1687.

[0063] Cetearyl alcohol polyether-20 was purchased from Guangzhou Xingye Technology Co., Ltd., item number: 003595-00288-5609.

[0064] Squalane was purchased from APRINNOVA, LLC, catalog number: 003432-05323-6673.

[0065] Cetearyl alcohol was purchased from Emery Oleochemicals (M) Sdn Bhd, catalog number: 003580-00664-1732.

[0066] Evaluation of antibacterial activity of herbal extracts in experimental cases and screening of synergistic acne-removing formulations 1. Experimental Objective This experiment aims to screen out single herbs with significant inhibitory effects on Propionibacterium acnes from 29 kinds of Chinese medicinal herbs with heat-clearing, detoxifying, dampness-drying, and itch-relieving effects using the inhibition zone diffusion method. Based on this, further optimization of compound formulations will be carried out to screen out Chinese medicine compositions with the best antibacterial effect and obvious synergistic effect, thereby providing a material basis for the development of safe and effective acne treatment products.

[0067] 2. Experimental Principle Propionibacterium acnes is one of the key pathogens in the pathogenesis of acne. This bacterium typically colonizes the hair follicles and sebaceous glands of the skin as a symbiotic organism. When the hair follicle opening is blocked and sebum secretion is excessive, it can proliferate rapidly in an anaerobic environment. Its metabolic byproducts, such as free fatty acids, can induce and aggravate local inflammatory responses, leading to acne lesions such as comedones, papules, and pustules. Therefore, inhibiting the proliferation of Propionibacterium acnes is an important strategy for the prevention and treatment of acne.

[0068] The inhibition zone diffusion method (disk method) is a classic method for evaluating the antibacterial activity of drugs in vitro. Its principle involves placing a sterile paper disc soaked in the test sample onto the surface of a solid agar medium evenly inoculated with specific bacteria. The antibacterial components in the test sample diffuse outwards through the agar, forming a concentration gradient from high to low. Within the effective concentration range of the drug, bacterial growth is inhibited, thus forming a transparent sterile area around the paper disc, known as the "zone of inhibition." The diameter of the zone of inhibition is positively correlated with the antibacterial strength of the test sample. This method is simple to operate and provides intuitive results, making it suitable for rapid and preliminary activity screening of large numbers of samples.

[0069] 3. Instruments and Materials The main equipment used in the experiment included: KQ-500DE ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.), DHP-9082 constant temperature incubator (Shanghai Yiheng Scientific Instrument Co., Ltd.), ESCO BSC-1300IIA2 biosafety cabinet, Eppendorf micropipettes, MLS-3781 autoclave (SANYO), and vernier calipers.

[0070] Medicinal materials and reagents: A total of 29 kinds of medicinal materials were used for testing, all of which were commercially available Chinese herbal medicine slices purchased from Guangzhou Xiwotang Trading Co., Ltd. They were verified to meet the relevant standards in the Chinese Pharmacopoeia (2020 edition). Before use, the medicinal materials were dried to constant weight at 60℃, pulverized, and passed through a 40-mesh sieve. A detailed list is shown in Table 1. Extraction solvents included deionized water, 75% ethanol, propylene glycol, butylene glycol, and cosmetic-grade sweet almond oil. The culture medium was reinforced clostridium agar (RCA). Sterile filter paper discs (6 mm in diameter), sterile petri dishes (90 mm), sterile cotton swabs, sterile centrifuge tubes, pipette tips, etc.

[0071] Experimental strain: Standard strain Propionibacterium acnes (ATCC 6919), purchased from Guangdong Institute of Microbiology.

[0072] 4. Experimental Methods 4.1 Preparation of single-herb extract solutions Accurately weigh 1.0 g each of 29 kinds of dried, pulverized, and sieved medicinal materials (passed through a 40-mesh sieve) and place them separately into 50 mL stoppered conical flasks. Add 10 mL of extraction solvent (deionized water, 75% ethanol, propylene glycol, butylene glycol, or sweet almond oil) to each flask, making the solid-liquid ratio (w / v) 1:10. Place the conical flasks in an ultrasonic cleaner and extract ultrasonically at 30℃ and 400 W for 60 minutes. After extraction, centrifuge the extract at 15000 r / min for 10 minutes, collect the supernatant, and filter it through a 0.45 μm microporous membrane to obtain the single-herb extract solution of each medicinal material. Store the solution at 4℃ for later use in screening for antibacterial activity.

[0073] 4.2 Initial screening of single-component antibacterial activity Propionibacterium acnes was inoculated onto reinforced Clostridium agar (RCA) plates and incubated at 37°C in an anaerobic environment (85% N2, 10% H2, 5% CO2) for 48 hours. Single colonies were picked and the bacterial concentration was adjusted to 0.5 McFarland turbidity (approximately 1.5 × 10⁻⁶) with sterile physiological saline. 8 (CFU / mL).

[0074] Spread 100 μL of bacterial suspension evenly onto the surface of a solidified RCA agar plate. Immediately transfer the plate to an anaerobic environment at 37°C and let it stand for 15–20 minutes to allow the bacteria to fully absorb the suspension. Take a sterile filter paper disc (6 mm in diameter), absorb 10 μL of extract, evaporate the solvent at room temperature, and immediately affix it to the bacterial plate. Incubate the plate in an anaerobic environment (85% N2, 10% H2, 5% CO2) at 37°C for 48 hours.

[0075] After incubation, the diameter of the inhibition zone (including the diameter of the filter paper) was measured using calipers. Four replicates were prepared for each extract, and the average result was taken. An inhibition zone diameter ≥10 mm was used as the screening threshold for significant antibacterial activity.

[0076] 4.3 Screening for Synergistic Effects of Compound Drugs Based on the initial screening results of single-component herbs, medicinal materials with significant inhibitory effects on Propionibacterium acnes (inhibition zone diameter ≥10 mm) and their corresponding optimal extraction solvents were selected. Multiple binary, ternary and quaternary compound combinations (numbered F-01 to F-57) were designed and prepared to screen for compatibility with synergistic effects.

[0077] Preparation of compound extracts: Each herb was extracted, centrifuged, and filtered independently according to the method in "4.1 Herb Extraction" to obtain extracts of each single herb. To examine the synergistic effect of different herb combinations, the extracts of each single herb were mixed in equal volume proportions when preparing the compound, and the concentration of crude drug in each extract was converted to ensure that the total crude drug concentration of the compound extract was consistent with the crude drug concentration of the extract used in the initial screening of the single herbs, thus making the antibacterial activity evaluation results of the single herbs and the compound comparable.

[0078] Evaluation of the antibacterial activity of the compound: The antibacterial activity of the compound was evaluated using the same filter paper agar diffusion method as in "4.2 Initial Screening of Antibacterial Activity of Single Herbs". By systematically comparing the diameter of the inhibition zone of each compound with its constituent single herbs, the changes in antibacterial activity after compatibility were analyzed, thereby screening out specific herbal combinations with the best antibacterial activity and a clear synergistic enhancement effect.

[0079] 5. Experimental Results 5.1 Initial screening results of single-component antibacterial activity Following the method described in section 4.1, 29 medicinal materials were subjected to ultrasonic extraction using five solvents: deionized water, 75% ethanol, propylene glycol, butylene glycol, and sweet almond oil, resulting in 145 single-herb extract samples. Using an inhibition zone diameter ≥10 mm as the screening criterion for significant antibacterial activity, the results showed that 6 extracts met this threshold. Specific data are shown in Table 1.

[0080] Table 1. Diameter of inhibition zone (mm) of single extracts against Propionibacterium acnes ±s, n=4)

[0081] Note: * indicates that the screening threshold (≥10 mm) has been reached.

[0082] The results in Table 1 show that six extracts—Polygonum cuspidatum almond oil extract, Cyprinus cuspidatum almond oil extract, Myrtle almond oil extract, Portulaca oleracea ethanol extract, Salvia miltiorrhiza almond oil extract, and Dictamnus dasycarpus propylene glycol extract—had inhibition zones of 10 mm or more against Propionibacterium acnes and were identified as potential single extracts for further screening of compound combinations.

[0083] 5.2 Results of screening for synergistic effects of compound drugs Based on the initial screening results of the single herbs mentioned above, multiple compound formulations were designed and inhibition zone tests were conducted. The results showed that most compound formulations exhibited better inhibitory effects against *Propionibacterium acnes* than their individual single herb extracts. Among them, the four-herb compound (combination No. 44) composed of *Rhodomyrtus tomentosa* sweet almond oil extract, *Polygonum cuspidatum* sweet almond oil extract, *Salvia miltiorrhiza* sweet almond oil extract, and *Dictamnus dasycarpus* propylene glycol extract showed the best antibacterial effect, with an inhibition zone diameter of 17.6 ± 0.7 mm. This value is significantly larger than the inhibition zone diameter of each single herb extract (the maximum single-herb diameter was 14.3 mm) and also higher than all other tested compound formulations. Specific data are shown in Table 2.

[0084] Table 2. Diameter of the inhibition zone (mm) of the compound combination against Propionibacterium acnes ±s, n=4)

[0085] 6. Conclusion This experiment targeted Propionibacterium acnes and used the in vitro inhibition zone diffusion method to screen 29 kinds of traditional Chinese medicines with heat-clearing, detoxifying and dampness-drying properties for single-herb formulation and optimize compound formulations. The aim was to obtain traditional Chinese medicine compositions with excellent antibacterial activity and significant synergistic effects, providing experimental basis for the development of acne-removing functional raw materials.

[0086] The results of single-herb screening showed that six extracts—Polygonum cuspidatum, Cyprinus dasycarpus, Myrtle, sweet almond oil extract of Salvia miltiorrhiza, ethanol extract of Portulaca oleracea, and propylene glycol extract of Dictamnus dasycarpus—all exhibited inhibition zones ≥10 mm in diameter against Propionibacterium acnes and were identified as potential single-herb extracts with significant antibacterial activity. Extracts from other herbs and solvent combinations did not meet the screening threshold, indicating that the compatibility between the extraction solvent and the active ingredients of the herbs has a crucial impact on the antibacterial effect.

[0087] The results of compound screening showed that the antibacterial activities of different herbal combinations varied significantly. Among them, the four-component combination (combination F-44) consisting of sweet almond oil extracts of Myrtle, Polygonum cuspidatum, and Salvia miltiorrhiza, and propylene glycol extract of Dictamnus dasycarpus root bark exhibited the best antibacterial effect, with an inhibition zone diameter of 17.6 ± 0.7 mm against Propionibacterium acnes, significantly higher than that of each single herb extract (maximum inhibition zone diameter of 14.3 mm) and other compound combinations, demonstrating a clear synergistic effect. This indicates that the combination of these four herbs is not a simple superposition of active ingredients, but rather achieves a significant enhancement of antibacterial activity through synergistic interaction between components. Based on this, this combination was identified as the traditional Chinese medicine composition of this invention.

[0088] In summary, the optimized traditional Chinese medicine composition and its suitable stepwise solvent extraction process selected in this experiment provide experimental basis for developing low-irritation, high-efficacy natural acne-removing preparations.

[0089] Example 1: Preparation of Traditional Chinese Medicine Composition (1) Preparation of various extracts: Preparation method of myrtle extract: It is prepared by ultrasonic-assisted extraction. Sweet almond oil is used as solvent. Myrtle powder is added at a material-to-liquid ratio of 1:10 (herb weight g: solvent volume mL, w / v). It is extracted for 60 minutes at 400 W ultrasonic power and 30℃. After removing the residue by coarse filtration, the extract is centrifuged at 15000 r / min using a tubular centrifuge. The supernatant obtained is myrtle extract.

[0090] The preparation methods for Polygonum cuspidatum extract and Salvia miltiorrhiza extract are the same as those for Myrtle extract.

[0091] Preparation method of Dictamnus dasycarpus extract: It is prepared by ultrasonic-assisted extraction. Propylene glycol is used as solvent. The crude powder of Dictamnus dasycarpus is added at a material-to-liquid ratio of 1:10 (medicinal material weight g: solvent volume mL, w / v). The extraction is carried out at 400 W ultrasonic power and 30℃ for 60 minutes. After removing the residue by coarse filtration, the extract is centrifuged at 15000 r / min using a tubular centrifuge. The supernatant obtained is the Dictamnus dasycarpus extract.

[0092] (2) Preparation of the composition: The extracts of Myrtle, Polygonum cuspidatum, Salvia miltiorrhiza and Dictamnus dasycarpus obtained in the above steps are mixed in a mass ratio of 3:3:1.5:1.5 and dispersed by mechanical stirring to obtain the traditional Chinese medicine composition of Example 1.

[0093] Example 2 Preparation of Traditional Chinese Medicine Composition The preparation methods for the various extracts are the same as in Example 1, except that the mass ratio of the myrtle extract, Japanese knotweed extract, salvia miltiorrhiza extract and dictamnus dasycarpus extract is 4:3:1:1.5 when the composition is prepared.

[0094] Example 3 Preparation of Traditional Chinese Medicine Composition The preparation methods for the various extracts are the same as in Example 1, except that the mass ratio of the myrtle extract, Japanese knotweed extract, salvia miltiorrhiza extract and dictamnus dasycarpus extract is 3:2:2:2 when the composition is prepared.

[0095] Example 4: Preparation of Traditional Chinese Medicine Composition The preparation methods for the various extracts are the same as in Example 1, except that the mass ratio of the mixed extracts of Myrtle Extract, Polygonum cuspidatum Extract, Salvia miltiorrhiza Extract and Dictamnus dasycarpus Extract is 2:3:1.5:2.

[0096] Example 5 Preparation of Traditional Chinese Medicine Composition The preparation methods for the various extracts are the same as in Example 1, except that the mass ratio of the mixed extracts of Myrtle Extract, Polygonum cuspidatum Extract, Salvia miltiorrhiza Extract and Dictamnus dasycarpus Extract is 4:2:1.5:1.

[0097] Example 6 Typical Product Dosage Form: Preparation of Acne-Clearing Gel This embodiment provides a method for preparing an acne-removing gel containing the traditional Chinese medicine composition of the present invention. The formulation composition is shown in Table 3.

[0098] Table 3. Composition of Acne Gel Formula (per 100 parts by weight)

[0099] Preparation method: Take an appropriate amount of deionized water, add 0.5 parts of carbomer 940, let it stand to swell, then stir to disperse evenly. Add 5.0 parts of glycerol and stir until completely dissolved. Separately, mix 3.0 parts of PEG-40 hydrogenated castor oil with 3.0 parts of the traditional Chinese medicine composition prepared in Example 1, and stir evenly to completely disperse the oil-soluble components, forming a clear or slightly turbid solubilized phase. Under stirring conditions, slowly add the solubilized phase to the above aqueous phase, and continue stirring until the system is evenly dispersed. Then slowly add an appropriate amount of triethanolamine while stirring, and adjust the pH to 6.0–7.0. The system gradually forms a transparent or translucent gel. Dissolve 0.5 parts of p-hydroxyacetophenone and 0.5 parts of 1,2-hexanediol by preheating to 50°C, add them to the gel, stir evenly, add deionized water to 100 parts, and continue stirring until the system is homogeneous, fine, and free of bubbles.

[0100] Example 7 Typical Product Dosage Form: Preparation of Acne Treatment Essence This embodiment provides a method for preparing an acne-removing essence containing the traditional Chinese medicine composition of the present invention. The formula composition is shown in Table 4.

[0101] Table 4. Composition of Acne Treatment Serum Formula (per 100 parts by weight)

[0102] Preparation method: Take an appropriate amount of deionized water, add 0.1 parts of sodium hyaluronate, and stir until it swells completely and is free of particles. Then add 10.0 parts of 1,3-butanediol and 3.0 parts of glycerol, and stir until completely dissolved to form a homogeneous aqueous solution. Separately, mix 3.0 parts of PEG-40 hydrogenated castor oil with 3.0 parts of the traditional Chinese medicine composition prepared in Example 1, and stir evenly to completely disperse the oil-soluble active ingredients, forming a clear or slightly turbid solubilized phase. Under continuous stirring, slowly add the solubilized phase to the aqueous phase, stirring until the system is completely homogeneous and there is no layering. Then add 0.5 parts of p-hydroxyacetophenone and 0.5 parts of 1,2-hexanediol, and continue stirring until fully dissolved. Add deionized water to a final volume of 100 parts, stir evenly, and allow to stand to defoam, thus obtaining the final product.

[0103] Example 8 Typical Product Dosage Form: Preparation of Acne Cream This embodiment provides a method for preparing an acne-removing cream containing the traditional Chinese medicine composition of the present invention. The formulation composition is shown in Table 5.

[0104] Table 5. Composition of Acne Cream Formula (per 100 parts by weight)

[0105] Preparation method: Take 70.0 parts of deionized water, 5.0 parts of glycerin, 0.5 parts of 1,2-hexanediol, and 0.5 parts of p-hydroxyacetophenone, place them in an aqueous phase container, heat to 75°C, and stir until completely dissolved to obtain the aqueous phase. Take 3.0 parts of the traditional Chinese medicine composition prepared in Example 1, 10.0 parts of squalane, 3.0 parts of cetearyl alcohol polyether-20, and 8.0 parts of cetearyl alcohol, place them in an oil phase container, heat to 75°C, and stir until all components are completely melted and mixed evenly to obtain the oil phase. Under stirring conditions, slowly add the oil phase to the aqueous phase, maintain the temperature at 75°C, and homogenize at high speed (3000 r / min) for 5 minutes until a uniform and fine milky white cream matrix is ​​formed. Allow to cool naturally to room temperature under continuous stirring, and continue stirring until the system is uniform and smooth to obtain the O / W type acne cream.

[0106] Comparative Example 1: Preparation of Traditional Chinese Medicine Composition (lacking Myrtle) The preparation methods for the various extracts are the same as in Example 1, except that myrtle extract is not added when preparing the composition, and the other three extracts are mixed in a mass ratio of Polygonum cuspidatum extract: Salvia miltiorrhiza extract: Dictamnus dasycarpus extract = 3:1.5:1.5.

[0107] Comparative Example 2: Preparation of Traditional Chinese Medicine Composition (lacking Polygonum cuspidatum) The preparation methods for the various extracts are the same as in Example 1, except that Polygonum cuspidatum extract is not added when preparing the composition, and the other three extracts are mixed in a mass ratio of Myrtle extract: Salvia miltiorrhiza extract: Dictamnus dasycarpus extract = 3:1.5:1.5.

[0108] Comparative Example 3: Preparation of Traditional Chinese Medicine Composition (without Danshen) The preparation methods for various extracts are the same as in Example 1, except that salvia miltiorrhiza extract is not added when preparing the composition, and the other three extracts are mixed in a mass ratio of myrtle extract: Polygonum cuspidatum extract: Dictamnus dasycarpus extract = 3:3:1.5.

[0109] Comparative Example 4: Preparation of Traditional Chinese Medicine Composition (without Dictamnus dasycarpus root bark) The preparation methods for the various extracts are the same as in Example 1, except that Dictamnus dasycarpus extract is not added when preparing the composition, and the other three extracts are mixed in a mass ratio of Myrtle extract: Polygonum cuspidatum extract: Salvia miltiorrhiza extract = 3:3:1.5.

[0110] Comparative Example 5: Preparation of Traditional Chinese Medicine Composition (lacking Myrtle and Polygonum cuspidatum) The preparation methods for various extracts are the same as in Example 1, except that myrtle extract and Polygonum cuspidatum extract are not added when preparing the composition, and the other two extracts are mixed at a mass ratio of Salvia miltiorrhiza extract: Dictamnus dasycarpus extract = 1.5:1.5.

[0111] Comparative Example 6: Preparation of Traditional Chinese Medicine Composition (lacking Polygonum cuspidatum and Salvia miltiorrhiza) The preparation methods for various extracts are the same as in Example 1, except that Polygonum cuspidatum extract and Salvia miltiorrhiza extract are not added when preparing the composition, and the other two extracts are mixed at a mass ratio of Myrtle extract: Dictamnus dasycarpus extract = 3:1.5.

[0112] Comparative Example 7: Preparation of Traditional Chinese Medicine Composition (lacking Myrtle and Salvia miltiorrhiza) The preparation methods for various extracts are the same as in Example 1, except that myrtle extract and salvia miltiorrhiza extract are not added when preparing the composition, and the other two extracts are mixed at a mass ratio of Polygonum cuspidatum extract: Dictamnus dasycarpus extract = 3:1.5.

[0113] Comparative Example 8: Preparation of Traditional Chinese Medicine Composition (lacking Salvia miltiorrhiza and Dictamnus dasycarpus root bark) The preparation methods for various extracts are the same as in Example 1, except that salvia miltiorrhiza extract and dictamnus dasycarpus extract are not added when preparing the composition, and the other two extracts are mixed at a mass ratio of myrtle extract: Polygonum cuspidatum extract = 3:3.

[0114] Comparative Example 9: Preparation of Traditional Chinese Medicine Composition (Full Water Extraction) The preparation methods for various extracts are basically the same as in Example 1, except that deionized water is used as the only solvent, while the other operations remain unchanged. Finally, the extracts of Myrtle, Polygonum cuspidatum, Salvia miltiorrhiza and Dictamnus dasycarpus are mixed in a mass ratio of 3:3:1.5:1.5.

[0115] Comparative Example 10: Preparation of Traditional Chinese Medicine Composition (All-Ethanol Extraction) The preparation methods of various extracts are basically the same as those in Example 1, except that 75% ethanol is used as the only solvent, while the other operations remain unchanged. Finally, the extracts of myrtle, Polygonum cuspidatum, Salvia miltiorrhiza and Dictamnus dasycarpus are mixed in a mass ratio of 3:3:1.5:1.5.

[0116] Comparative Example 11: Preparation of Traditional Chinese Medicine Composition (All-Propylene Glycol Extract) The preparation methods of various extracts are basically the same as those in Example 1, except that propylene glycol is used as the only solvent, while the other operations remain unchanged. Finally, the extracts of myrtle, Polygonum cuspidatum, Salvia miltiorrhiza and Dictamnus dasycarpus are mixed in a mass ratio of 3:3:1.5:1.5.

[0117] Comparative Example 12: Preparation of Traditional Chinese Medicine Composition (Full Oil Extraction) The preparation methods for various extracts are basically the same as in Example 1, except that sweet almond oil is used as the only solvent, while the other operations remain unchanged. Finally, the myrtle extract, Japanese knotweed extract, salvia miltiorrhiza extract and dictamnus dasycarpus root bark extract are mixed in a mass ratio of 3:3:1.5:1.5.

[0118] Comparative Example 13: Preparation of Traditional Chinese Medicine Composition (Extraction Solvent Replacement - Polygonum cuspidatum) The preparation methods of various extracts are basically the same as those in Example 1, except that the Polygonum cuspidatum extract is extracted with 75% ethanol by ultrasound. The other operations remain unchanged. Finally, the extracts of Myrtle, Polygonum cuspidatum, Salvia miltiorrhiza and Dictamnus dasycarpus are mixed in a mass ratio of 3:3:1.5:1.5.

[0119] Comparative Example 14: Preparation of Traditional Chinese Medicine Composition (Extraction Solvent Replacement - Danshen) The preparation methods of various extracts are basically the same as those in Example 1, except that the Salvia miltiorrhiza extract is extracted with 75% ethanol by ultrasound. The other operations remain the same. Finally, the extracts of Myrtle, Polygonum cuspidatum, Salvia miltiorrhiza and Dictamnus dasycarpus are mixed in a mass ratio of 3:3:1.5:1.5.

[0120] Comparative Example 15: Preparation of Traditional Chinese Medicine Composition (Formulation Exceeds Patent Scope) The preparation methods for various extracts are the same as in Example 1, except that in the formulation of the composition, the mass ratio of the mixed extracts of myrtle extract, Japanese knotweed extract, salvia miltiorrhiza extract and dictamnus dasycarpus extract is 5:3:1.5:1.5 (the proportion of myrtle extract is too high).

[0121] Comparative Example 16: Preparation of Traditional Chinese Medicine Composition (Formulation Exceeds Patent Scope) The preparation methods for various extracts are the same as in Example 1, except that in the formulation of the composition, the mass ratio of the mixed extracts of Myrtle Extract, Polygonum cuspidatum Extract, Salvia miltiorrhiza Extract and Dictamnus dasycarpus Extract is 3:5:1.5:1.5 (the proportion of Polygonum cuspidatum Extract is too high).

[0122] Comparative Example 17: Preparation of Traditional Chinese Medicine Composition (Formulation Exceeds Patent Scope) The preparation methods for various extracts are the same as in Example 1, except that in the formulation of the composition, the mass ratio of the mixed extracts of Myrtle Extract, Polygonum cuspidatum Extract, Salvia miltiorrhiza Extract and Dictamnus dasycarpus Extract is 3:3:3:1 (the proportion of Salvia miltiorrhiza Extract is too high).

[0123] Comparative Example 18: Preparation of Traditional Chinese Medicine Composition (Formulation Exceeds Patent Scope) The preparation methods for various extracts are the same as in Example 1, except that in the formulation of the composition, the mass ratio of the mixed extracts of Myrtle Extract, Polygonum cuspidatum Extract, Salvia miltiorrhiza Extract and Dictamnus dasycarpus Extract is 3:3:1:3 (the proportion of Dictamnus dasycarpus Extract is too high).

[0124] Test Example 1: Skin Irritation Test The methods and results of skin irritation tests on the traditional Chinese medicine compositions involved in Examples 1-5 are as follows: 1. Test method: The test was conducted in accordance with the "Skin Irritation Test Method" in the "Cosmetic Safety Technical Specifications (2015 Edition)".

[0125] 2. Experimental Principle: The test substance is applied once to the skin of the test animal. The degree of local skin irritation is observed and scored at specified time intervals. A self-control is used to evaluate the irritant effect of the test substance on the skin.

[0126] 3. Test Samples and Animals: The test substance was the acne-relieving and soothing composition described in Examples 1-5. No dilution is required; use directly at the original concentration, and shake well before use. The test animals were ordinary New Zealand white rabbits, purchased from the Huibei Experimental Animal Farm in Baiyun District, Guangzhou.

[0127] 4. Experimental steps: 4.1 Fifteen healthy New Zealand rabbits were randomly divided into five groups of three rabbits each, corresponding to samples from Examples 1 to 5.

[0128] 4.2 24 hours before the experiment, shave the hair on both sides of the spine on the back of the rabbit (about 3 cm × 3 cm on each side), taking care to avoid damaging the epidermis.

[0129] 4.3 Apply approximately 0.5 mL of the test solution evenly to one side of shaved skin, cover with two layers of gauze (2.5 cm × 2.5 cm) and one layer of cellophane, and secure with non-irritating adhesive tape and bandage for a occlusion test. The other side of skin serves as a blank control. The application time is 4 hours. After the test, remove any remaining test solution with warm water.

[0130] 4.4 Observe the skin reaction at the application site at 1, 24, 48, and 72 hours after removing the test substance, and score it according to Table 6. Based on the average of the highest scores at each time point of 24, 48, and 72 hours, assess the skin irritation intensity according to Table 7.

[0131] Table 6 Skin Irritation Response Scores

[0132] Table 7 Skin Irritation Intensity Grading

[0133] 5. Results and Analysis Within 72 hours of administration of the herbal compositions of the various embodiments of the present invention, no erythema or edema was observed on the skin of New Zealand rabbits. According to the skin irritation intensity grading in Table 7, it is demonstrated that the herbal compositions of the various embodiments of the present invention are non-irritating to the skin.

[0134] Test Example 2: Determination of the in vitro antibacterial activity of a traditional Chinese medicine composition against acne-associated pathogens. 1. Experimental Objective This experiment aims to determine the in vitro antibacterial activity of the herbal compositions of this invention (Examples 1-5) and control samples against three representative pathogenic bacteria closely related to the pathogenesis of acne using the filter paper diffusion method. By systematically evaluating the inhibitory effects of these compositions on the three bacteria, comprehensive microbiological experimental evidence is provided for their acne-removing and antibacterial efficacy.

[0135] 2. Experimental Principle This experiment selected three representative Gram-positive bacteria—Propionibacterium acnes, Staphylococcus aureus, and Staphylococcus epidermidis—to comprehensively cover the key microbial links in the pathogenesis of acne.

[0136] Propionibacterium acnes, as the core pathogenic bacterium of acne, directly induces inflammation and skin lesions through its proliferation, making it the primary bacterial species for evaluating acne treatment efficacy. Staphylococcus aureus, as a common secondary skin infection bacterium, easily leads to acne suppuration and exacerbation of inflammation; examining its inhibitory effect can verify the product's ability to prevent complications. Although Staphylococcus epidermidis is a normal skin flora, it can transform into an opportunistic pathogen under certain conditions and participate in biofilm formation, affecting the microecological balance of acne and the difficulty of treatment.

[0137] These three bacteria cover the entire chain from primary acne to secondary infection, which fully demonstrates the antibacterial activity (acne treatment) and clinical applicability of the composition of the present invention in the complex skin microenvironment.

[0138] The filter paper diffusion method is based on the formation of a concentration gradient by the diffusion of antibacterial substances in agar medium, thereby inhibiting bacterial growth. Sterile filter paper discs soaked in the test sample are placed on the surface of an agar plate inoculated with bacterial solution. After incubation, the diameter of the inhibition zone is measured; its size is positively correlated with the antibacterial strength of the test sample. This method is simple to operate and provides intuitive results.

[0139] 3. Test sample and reference sample 3.1 Bacterial strains and culture media The strains used in the experiment were provided by the Guangdong Institute of Microbiology, including Propionibacterium acnes (ATCC 6919), Staphylococcus aureus (ATCC 25923), and Staphylococcus epidermidis (ATCC 12228).

[0140] Culture media were selected according to the characteristics of the bacterial species: Mueller-Hinton agar (MHA) was used for aerobic bacteria (Staphylococcus aureus, Staphylococcus epidermidis); and reinforced Clostridium agar (RCA) was used for anaerobic bacteria (Propionibacterium acnes). Cultures were placed in an anaerobic environment (85% N2, 10% H2, 5% CO2). Bacterial suspensions were prepared with sterile physiological saline (0.85% NaCl) and adjusted to a McFarland turbidity of 0.5 (approximately 1.5 × 10⁻⁶). 8 (CFU / mL).

[0141] 3.2 Test Sample and Reference Sample Test sample group: Example group (1-5) is the core formulation of this invention, a traditional Chinese medicine composition prepared according to the method of Example 1-5. Comparative example group (1-18) is a defect or process control formulation, prepared according to the corresponding comparative example method.

[0142] Control group: The positive control was chloramphenicol antibiotic tablets (30 μg / tablet, catalog number OXOID CT0014B). The negative control was a blank matrix solution containing no active ingredient (solvent system was the same as in Example 1).

[0143] 4. Instruments, equipment and consumables The equipment included a laminar flow hood (SW-CJ-2FD, Suzhou Jingantai), a constant temperature incubator (DHP-9272, Shanghai Yiheng), a McFarland turbidimeter (DensiCHEK Plus, bioMérieux, USA), an anaerobic incubator (with gas-generating bag, Mitsubishi), and vernier calipers. Experimental consumables included sterile filter paper (6 mm diameter), sterile petri dishes, sterile cotton swabs, and pipettes.

[0144] 5. Experimental Methods 5.1 Preparation of test sample Test sample group: Take the original solutions of the traditional Chinese medicine compositions prepared in Examples 1-5 and Comparative Examples 1-18 respectively, and dilute or adjust them with the corresponding solvent system according to the total amount of raw medicinal materials in each composition during preparation (for Examples 1-8 and Comparative Examples 13-18, sweet almond oil and propylene glycol were mixed in the corresponding proportions, and for Comparative Examples 9-12, water, 75% ethanol, propylene glycol and sweet almond oil were used respectively) so that the total concentration of raw medicinal materials in each test sample solution is uniformly 0.10 g / mL (i.e., each milliliter of solution is equivalent to 0.10 g of raw medicinal materials), and use it as the working solution for antibacterial activity testing.

[0145] Positive control group: Chloramphenicol antibiotic tablets (30 μg / tablet).

[0146] Negative control group: blank matrix solution without active ingredients (the solvent system is the same as in Example 1, and the solvent ratio is adjusted to be equivalent to that of the test sample group in the same way).

[0147] 5.2 Preparation of bacterial suspension Standard strains of Propionibacterium acnes, Staphylococcus aureus, and Staphylococcus epidermidis were inoculated into their respective solid culture media for resuscitation. Propionibacterium acnes was cultured anaerobically at 37°C for 48 hours, while Staphylococcus aureus and Staphylococcus epidermidis were cultured at 37°C for 24 hours. Single colonies were picked and the bacterial concentration was adjusted to 0.5 McFarland turbidity (approximately 1.5 × 10⁻⁶) with sterile physiological saline. 8 (CFU / mL), used as a bacterial suspension for experiments.

[0148] 5.3 Antibacterial activity assay Melt and cool the corresponding agar medium (MHA for Staphylococcus aureus and Staphylococcus epidermidis, and RCA for Propionibacterium acnes) to approximately 45°C into sterile petri dishes, about 20 mL per dish, and allow it to solidify. Take 100 μL of the above bacterial suspension and spread it evenly on the surface of the solidified agar plate using a sterile spreader.

[0149] Take sterile filter paper discs (6 mm in diameter), and respectively aspirate 10 μL of the extracts from Examples 1–5, Comparative Examples 1–18, and the negative control (blank matrix solution). After evaporating the solvent at room temperature, affix them to bacterial plates. Plates containing Propionibacterium acnes are incubated in an anaerobic environment at 37°C for 48 hours. Plates containing Staphylococcus aureus and Staphylococcus epidermidis are incubated in a constant temperature incubator at 37°C for 24 hours. Positive control chloramphenicol antibiotic discs are directly affixed to the corresponding bacterial plates.

[0150] After incubation, the diameter of the inhibition zone (including the diameter of the filter paper) was measured using calipers. Four replicates were prepared for each extract, and the average value was taken.

[0151] 5.4 Criteria for Determining Antibacterial Activity Referring to the relevant standards of the Clinical Laboratory Standards Institute (CLSI) and the evaluation conventions for the antibacterial efficacy of cosmetics, the following judgment criteria were set: A zone diameter ≤ 6 mm (paper diameter) indicates no antibacterial effect; 6 mm < zone diameter ≤ 10 mm indicates low sensitivity; 10 mm < zone diameter ≤ 15 mm indicates moderate sensitivity; and a zone diameter > 15 mm indicates high sensitivity. Furthermore, if the diameter of the inhibition zone of the test sample is significantly larger than that of the negative control group (P < 0.05) and ≥ 7 mm, then the test sample is considered to have antibacterial effect against the tested strain.

[0152] 6. Results and analysis of antibacterial activity 6.1 Results of inhibition zone diameter measurement The diameters of the inhibition zones of each test sample against Staphylococcus aureus, Staphylococcus epidermidis, and Propionibacterium acnes were determined using the filter paper diffusion method. The results are detailed in Table 8. The inhibition zone diameter diagrams for Staphylococcus aureus, Staphylococcus epidermidis, and Propionibacterium acnes in Example 1 are shown in Table 8. Figure 1 , Figure 2 , Figure 3 .

[0153] Table 8. Results of inhibition zone diameter determination (mm) of different traditional Chinese medicine compositions against three acne-associated pathogens. ±s, n=4)

[0154] Note: SPSS 26.0 software was used for data analysis. Data for each group are expressed as mean ± standard deviation (SD). Mean ± s) is expressed as mean ± standard deviation. One-way ANOVA was used to compare differences between groups, combined with Tukey's method for post-hoc testing. The significance level was set at α = 0.05, with P < 0.05 considered statistically significant and P < 0.01 considered highly significant; the differences between each embodiment of this invention and all comparative examples were P < 0.01.

[0155] 6.2 Results Analysis The results of the filter paper diffusion method showed that the herbal composition of this invention exhibited significant inhibitory activity against three acne-associated pathogens. Specific analysis is as follows: (1) The composition has a strong and stable broad-spectrum antibacterial effect. As shown in Table 8, within the formulation range protected by the claims (Examples 1-5), the diameter of the inhibition zone of the composition against Staphylococcus aureus, Staphylococcus epidermidis, and Propionibacterium acnes is greater than 17 mm, which, according to the judgment criteria, constitutes "highly sensitive". Among them, the inhibitory activity against the core pathogen Propionibacterium acnes is particularly outstanding (inhibition zone 17.9-18.8 mm). The data differences between the examples are small, indicating that the antibacterial effect of the composition is stable within this formulation range.

[0156] (2) The specific combination of the four medicinal extracts produced a clear synergistic effect. The data comparison between the examples and the comparative examples (Comparative Examples 1-8) clearly demonstrates synergistic effects. For example, the inhibition zone against Propionibacterium acnes in Example 1 was 18.5 mm, while in the comparative examples lacking any one herb, the inhibition zone decreased to 13.8-15.2 mm; when two herbs were missing (Comparative Examples 5-8), the activity further weakened to 10.5-15.0 mm. This indicates that the specific combination of "Myrtle-Polygonum cuspidatum-Salvia miltiorrhiza-Dictamnus dasycarpus" produces a synergistic antibacterial effect of "1+1+1+1>4" through the interaction between components, rather than a simple additive effect.

[0157] (3) Adaptable stepwise solvent extraction process is the key to achieving efficient antibacterial effect. The results of the extraction process comparison showed that the compositions extracted with a single solvent (water, ethanol, propylene glycol, or sweet almond oil) (Comparative Examples 9-12) exhibited significantly lower antibacterial activity than those in the embodiments of this invention. In particular, changing the suitable solvent for each individual herb (e.g., replacing Polygonum cuspidatum or Salvia miltiorrhiza with alcohol extraction, see Comparative Examples 13-14) also significantly reduced the overall antibacterial effect. This indicates that selecting extraction solvents tailored to the characteristics of the active components of different herbs is the core of the process for ensuring the potent antibacterial activity of the compositions.

[0158] (4) A specific ratio range is the guarantee for maintaining optimal activity. Comparative Examples 15–18 (with proportions exceeding the scope of the claims) showed a reduction in the diameter of the inhibition zone against Propionibacterium acnes to 14.4–15.6 mm, a significant decrease compared to Example 1 (P<0.01). This indicates that the mass ratio range defined in the claims is optimized, within which the components achieve the best synergistic balance, while exceeding this range disrupts this balance, leading to a significant decrease in antibacterial activity.

[0159] (5) Comparison with positive control The positive control chloramphenicol (30 μg / tablet) showed a larger inhibition zone diameter (24.3–28.6 mm) against the three bacteria. Although the antibacterial activity of the herbal composition of this invention is lower than that of chemical antibiotics, its antibacterial effect has reached the expected level of cosmetic efficacy raw materials, but it has the advantages of low irritation and safety due to its natural herbal origin.

[0160] Test Example 3: Human Evaluation Experiment of Oil Control Efficacy 1. Experimental Objective Excessive sebum secretion is a core factor in the pathogenesis of acne. This experiment aims to quantitatively evaluate the inhibitory effect of the herbal compositions of this invention (Examples 1-5) on sebum secretion through human trials, thereby verifying their oil-controlling efficacy.

[0161] 2. Test sample and grouping Examples 1-5: Traditional Chinese medicine compositions prepared in Examples 1-5.

[0162] Blank control group: a blank matrix containing no active ingredients (i.e., a mixture of sweet almond oil and propylene glycol in a mass ratio of 5:1).

[0163] 3. Instruments and Equipment Skin oil analyzer (Sebumeter® SM815, Courage + Khazaka), constant temperature and humidity laboratory (22±2℃, 50±10% RH), electronic balance, etc.

[0164] 4. Experimental Design and Methods This experiment was a self-controlled pre- and post-treatment study. A total of 60 qualified subjects were recruited and randomly divided into 6 groups of 10 each, corresponding to samples from Examples 1-5 and a blank control group (containing only the matrix). The forehead was used as the fixed test site, and measurements were taken using the Sebumeter® skin oil analyzer.

[0165] Participants were aged 18–45 years, regardless of gender; self-reported as having oily or combination skin, with preliminary sebum testing confirming a basal sebum content ≥ 100 μg / cm² on the forehead area; signed informed consent, voluntarily participating and adhering to the experimental procedures; and had not used retinoic acid, corticosteroids, or other medications affecting sebum metabolism within the past 4 weeks; and did not have any active inflammatory skin diseases on their face (such as moderate to severe acne, seborrheic dermatitis, etc.).

[0166] Before the experiment, all participants were required to cleanse their faces and sit quietly to allow for acclimatization. First, the basal sebum content (D0) on the forehead of all participants before product use was measured using Sebumeter®, repeated three times and the average value was recorded. Subsequently, participants used the designated sample twice daily, morning and evening, for seven consecutive days, applying 0.5g evenly to the entire face each time and gently patting until absorbed. During this period, the use of any other oil-controlling, acne-reducing, or exfoliating products was prohibited. On day 7, the forehead sebum content was measured again under the same conditions (D7).

[0167] 5. Data Processing The sebum reduction rate after product use was calculated for each subject: Sebum reduction rate (%) = [(D0 value - D7 value) / D0 value] × 100%. Paired t-tests were used to analyze the differences between D0 and D7 within each group, and one-way ANOVA was used to compare the differences between the example group and the blank control group.

[0168] 6. Results and Analysis The results are shown in Table 9. After 7 days of continuous use, the sebum content on the forehead of all example groups (1-5) was significantly reduced compared with that before use (D0) (P<0.01), with a sebum reduction rate between 17.7% and 18.3%. The sebum content in the blank control group showed no significant change during the same period (reduction rate 0.2%). There was no statistically significant difference in oil control effect among the example groups (P>0.05).

[0169] Table 9 Results of sebum secretion assay in human trials ( (±s, n=10)

[0170] This experiment shows that the herbal composition of the present invention (Examples 1-5) can significantly reduce sebum secretion in the skin (reduction rate >17%, P<0.01) after continuous use for 7 days, demonstrating a clear oil-controlling effect. Within the range of the claimed proportions, the effects of each embodiment are stable and significantly superior to the blank matrix. This efficacy, combined with the antibacterial, anti-inflammatory, and soothing effects of the composition, constitutes the basis for its comprehensive effect in preventing and treating acne at multiple stages.

[0171] Test Example 4: Evaluation Experiment of Soothing Efficacy The soothing efficacy of Examples 1-5 and Comparative Examples 1-18 was tested.

[0172] 1. Experimental Objectives and Principles Hyaluronic acid is a key extracellular matrix component for maintaining the skin barrier and hydration. Hyaluronidase can degrade hyaluronic acid, thereby triggering or exacerbating skin inflammatory responses. Therefore, inhibiting hyaluronidase activity is an important in vitro indicator for evaluating the soothing and anti-inflammatory efficacy of topical products. This experiment assessed the soothing activity of a traditional Chinese medicine composition by measuring its inhibition rate against hyaluronidase.

[0173] 2. Experimental Methods The experiment was conducted according to the industry-standard method (HMC-W1-029). Hyaluronic acid was used as the substrate, and the enzymatic reaction was carried out at 37°C in an acetate buffer system at pH 5.6. The absorbance was measured at 528 nm using Ehrlich's reagent, and the hyaluronidase inhibition rate was calculated. Dipotassium glycyrrhizate solution was used as a positive control.

[0174] 3. Reagents: Hyaluronidase BR, 300U~500U / mg, purchased from Yuanye Biotechnology; all other reagents were of analytical grade.

[0175] 4. Experimental Procedure 4.1 Solution Preparation (1) Acetic acid buffer solution (pH 5.6): Take 1155 μL of glacial acetic acid, dilute it to 100 mL with ultrapure water, mix well, and take 4.8 mL of it as solution A. Weigh 2.72 g of sodium acetate crystals, dissolve them in ultrapure water, and make up to 100 mL. Mix well and take 45.2 mL of it as solution B. Mix solutions A and B, make up to 100 mL with ultrapure water, and mix thoroughly. Accurately measure its pH value, and adjust the pH value to 5.6 using solution A or B.

[0176] (2) Hyaluronidase solution: Weigh 0.0100g of hyaluronidase and place it in a beaker. Add 4mL of the prepared acetic acid buffer solution and let it dissolve completely.

[0177] (3) Sodium hyaluronate solution (0.5 mg / mL): Weigh 0.01 g of sodium hyaluronate into a beaker, add 20 mL of acetic acid buffer solution, and stir until completely dissolved.

[0178] (4) Ehrlich reagent: Weigh 0.8g of p-dimethylaminobenzaldehyde and dissolve it in a mixture of 15mL concentrated hydrochloric acid and 15mL anhydrous ethanol.

[0179] (5) Sodium carbonate solution (1.0 mol / L): Weigh 5.3 g of sodium carbonate, dissolve it in ultrapure water, and make up to 50 mL.

[0180] (6) Acetylacetone solution: Take 3.5 mL of acetylacetone and dissolve it in 50 mL of 1.0 mol / L sodium carbonate solution. This solution should be prepared fresh before use.

[0181] (7) Calcium chloride solution (0.25 mmol / L): Weigh 0.02775 g of calcium chloride, dissolve it in ultrapure water and make up to 1000 mL.

[0182] (8) Sodium hydroxide solution (0.4mol / L): Weigh 1.6g of sodium hydroxide, dissolve it in ultrapure water and make up to 100mL.

[0183] (9) Dipotassium glycyrrhizate solution: Weigh 3.000g of dipotassium glycyrrhizate, dissolve it in pure water and make up to 100mL.

[0184] (10) Traditional Chinese Medicine Composition Solution: Accurately weigh 1.000 g of each example and comparative sample, place them in a 1000 mL volumetric flask, add an acetate buffer solution (pH 5.6) containing 3.0% (v / v) PEG-40 hydrogenated castor oil, shake vigorously and sonicate until the sample is completely dispersed, dilute to the mark with the same buffer solution, and shake well to obtain a traditional Chinese medicine composition test solution with a concentration of 1.0 mg / mL. The final concentration of PEG-40 hydrogenated castor oil in all test solutions is uniformly 3.0% (v / v). At the same time, prepare the same solvent system without the sample as a blank control.

[0185] 4.2 Determination of Hyaluronidase Activity Inhibition Rate Examples 1-5 and Comparative Examples 1-18 were selected as samples, and dipotassium glycyrrhizate solution was used as a positive control for the following operations: First, mix 0.1 mL of 0.25 mmol / L calcium chloride solution and 0.5 mL of hyaluronidase solution, and incubate at 37°C for 20 minutes. Next, add 0.5 mL of sample solution and continue incubation at 37°C for another 20 minutes. Then, add 0.5 mL of sodium hyaluronate solution and incubate at 37°C for 30 minutes, followed by incubation at room temperature for 5 minutes. Then, add 0.1 mL of 0.4 mol / L sodium hydroxide solution and 0.5 mL of acetylacetone solution sequentially, and heat the mixture in a boiling water bath for 15 minutes. After heating, quickly cool with ice water for 5 minutes. Finally, add 1.0 mL of Ehrlich's reagent and dilute with 3.0 mL of anhydrous ethanol, incubate at room temperature for 30 minutes to allow for sufficient color development, and measure the absorbance at 528 nm using a spectrophotometer.

[0186] Calculate the hyaluronidase inhibition rate using the following formula.

[0187] Hyaluronidase inhibition rate (%) = [(AB) - (CD)] / (AB) × 100% In the formula: A represents the absorbance of the blank group, which consists of buffer solution, enzyme, and substrate. B represents the absorbance of the blank control group, which consists of buffer solution + substrate (without enzyme). C represents the absorbance of the sample group; the sample group refers to the sample test solution + enzyme + substrate. D represents the absorbance of the sample control group, which refers to the sample test solution + substrate (without enzyme).

[0188] 5. Results and Analysis The results of the inhibition rate of hyaluronidase in each group of samples are shown in Table 10.

[0189] Table 10 Results of the inhibition rate of hyaluronidase in each group of samples ( ±s, n=5)

[0190] As shown in Table 10, within the range of proportions defined in the claims (Examples 1-5), the composition inhibits hyaluronidase at a rate of 61.57% to 65.48%, which is comparable to the positive control dipotassium glycyrrhizate (65.66%), indicating that it has excellent soothing activity.

[0191] From the results of the comparative examples lacking certain ingredients, the hyaluronidase inhibition rates of the three comparative examples lacking Myrtle (Comparative Example 1, 48.25%), Polygonum cuspidatum (Comparative Example 2, 46.83%), and Salvia miltiorrhiza (Comparative Example 3, 44.92%) decreased by 13.32, 14.74, and 16.65 percentage points, respectively, compared to Example 1 (61.57%), showing significant differences. It is noteworthy that the inhibition rate of the group lacking Dictamnus dasycarpus (Comparative Example 4) was 62.87%, which was not significantly different from the example. This precisely illustrates that Myrtle, Polygonum cuspidatum, and Salvia miltiorrhiza are the main contributors to the soothing effect, while Dictamnus dasycarpus, although contributing little to soothing, is crucial for synergistic antibacterial effects (considering the results of Test Example 2). When two or more herbs were missing (Comparative Examples 5–8), the inhibition rate further decreased to 32.15%–41.26%, confirming that the complete combination of the four herbs is a necessary condition for achieving the synergistic "antibacterial + soothing" dual effects of this composition.

[0192] The comparison of extraction processes showed that the inhibition rates of the compositions extracted with a single solvent (Comparative Examples 9-12) were significantly lower than those of the Example Groups; changing the suitable solvent for the single herb (Comparative Examples 13-14) also resulted in a significant decrease in inhibition rate. The inhibition rate of the imbalanced groups (Comparative Examples 15-18) decreased to 47.28%-51.36%, which was also significantly lower than that of the Example Groups.

[0193] In summary, the herbal composition of this invention exhibits a significant inhibitory effect on hyaluronidase, with an inhibition rate >61%, comparable to the positive control dipotassium glycyrrhizate, indicating its excellent soothing and anti-inflammatory activity. This superior efficacy relies on the complete compatibility of the four medicinal herbs, a targeted stepwise solvent extraction process, and optimized specific quality ratios.

Claims

1. A traditional Chinese medicine composition with anti-acne soothing effect, characterized in that, The traditional Chinese medicine composition is composed of extracts of Myrtle, Polygonum cuspidatum, Salvia miltiorrhiza and Dictamnus dasycarpus in a mass ratio of (2-4):(2-4):(1-2):(1-2); the Myrtle, Polygonum cuspidatum and Salvia miltiorrhiza extracts are plant oil extracts obtained by using cosmetic-grade plant oils as extraction solvents; the Dictamnus dasycarpus extract is a propylene glycol extract obtained by using propylene glycol as extraction solvents.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The cosmetic-grade plant oils are selected from one or more of the following: sweet almond oil, peach kernel oil, olive oil, grape seed oil, jojoba oil, safflower seed oil, sunflower seed oil, avocado oil, and meadowfoam seed oil.

3. The traditional Chinese medicine composition according to claim 1, characterized in that, The cosmetic-grade plant oils are sweet almond oil, grapeseed oil, or jojoba oil.

4. The preparation method of the traditional Chinese medicine composition according to any one of claims 1-3, characterized in that, Includes the following steps: S1. Using cosmetic-grade plant oils as extraction solvents, extract the three medicinal herbs Myrtle, Polygonum cuspidatum, and Salvia miltiorrhiza, and obtain the corresponding plant oil extracts after centrifugation. S2. Using propylene glycol as the extraction solvent, extract the medicinal material of Dictamnus dasycarpus, and obtain the propylene glycol extract of Dictamnus dasycarpus after centrifugation and filtration. S3. Mix the extracts of Myrtle, Polygonum cuspidatum, Salvia miltiorrhiza and Dictamnus dasycarpus obtained in the above steps at a mass ratio of (2-4):(2-4):(1-2):(1-2) and disperse them to obtain the Chinese herbal composition.

5. The preparation method according to claim 4, characterized in that, In steps S1 and S2, the ratio of medicinal materials to extraction solvent is 1:10-15 (w / v).

6. The preparation method according to claim 4, characterized in that, The extraction described in steps S1 and S2 is ultrasonic extraction, with an extraction temperature of 25-50℃, an ultrasonic power of 400-500 W, and an extraction time of 60-90 minutes.

7. The use of the traditional Chinese medicine composition according to any one of claims 1 to 3 in the preparation of topical products for acne treatment, oil control, antibacterial and / or soothing.

8. An external use product having an anti-acne, oil control, antibacterial and / or soothing effect, characterized in that, The traditional Chinese medicine composition contains any one of the claims 1 to 3.

9. The product for external use according to claim 8, characterized by The mass content of the traditional Chinese medicine composition is 2-4%.

10. The use according to claim 7 or the external product according to claim 8, characterized in that, The external products mentioned are gels, serums, creams, masks, toners, ointments, or lotions.

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