A composition for treating acne and a method for preparing and using the same

Abstract

The present application relates to a kind of acne-removing composition and its preparation method and application, belong to the field of cosmetics technology.The acne-removing composition of the present application includes Gentiana straminea Maxim extract, biota orientalis leaf extract and norway spruce leaf extract, wherein Gentiana straminea Maxim extract is prepared by ultrasonic treatment after mixed bacteria fermentation of Gentiana straminea Maxim as raw material by Kluyveromyces marxianus and Bacillus pumilus, and the obtained Gentiana straminea Maxim extract has improved anti-inflammatory effect.The Gentiana straminea Maxim extract prepared in the present application is compounded with biota orientalis leaf extract and norway spruce leaf extract, and through determination, the three can synergistically improve the inhibitory effect of acne-removing composition on propionibacterium acnes, and simultaneously, the acne-removing composition of the present application can exert acne-removing effect from anti-inflammatory and bacteriostatic, oil control and other aspects, reduce facial acne area, reduce facial inflammation and reduce oil secretion.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, and in particular to an acne-removing composition, its preparation method, and its application. Background Technology

[0002] Acne is a common chronic inflammatory skin disease. The pathogenesis of acne involves the synergistic effects of multiple factors, including excessive sebum secretion, abnormal follicular keratinization, colonization of Propionibacterium acnes, and inflammatory response.

[0003] Currently, commercially available acne treatment products mainly rely on chemically synthesized ingredients such as salicylic acid, retinoic acid, and benzoyl peroxide. While these can suppress inflammation in the short term, long-term use can easily lead to side effects such as skin stinging, peeling, and barrier damage, and are not well-suited for sensitive skin. Therefore, natural plant extracts, due to their gentleness, multi-target synergistic effects, and low toxicity, have become an important direction for acne treatment product development. However, single plant extracts have limited targets, making it difficult to achieve full-chain regulation of oil control, antibacterial, anti-inflammatory, and repair. Existing compound compositions generally suffer from poor permeability of active ingredients, low bioavailability, and unclear synergistic mechanisms between different plant components, resulting in unstable acne treatment effects and long onset periods. Therefore, developing a multi-target synergistic, highly effective, and gentle plant extract composition has become an urgent technical problem to be solved in the field of acne treatment. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of the prior art and provide an acne-removing composition, its preparation method, and its application.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0006] In a first aspect, the present invention provides an acne-removing composition comprising the following components in parts by weight: 3-6 parts Gentiana macrophylla extract, 1-4 parts Platycladus orientalis leaf extract and 0.5-2 parts Norwegian spruce leaf extract; wherein the Gentiana macrophylla extract is prepared by using Gentiana macrophylla as raw material, through mixed fermentation of Kluyveromyces martensii and Bacillus pumilus, followed by ultrasonic treatment.

[0007] This invention discovers that mixed fermentation of Gentiana macrophylla with specific strains and dosage ratios of Kluyveromyces martensii and Bacillus pumilus significantly enhances the inhibitory effect of Gentiana macrophylla extract on inflammatory factors TNF-α and IL-6. This indicates that mixed fermentation of Gentiana macrophylla with specific strains and viable cell ratios of Kluyveromyces martensii and Bacillus pumilus can improve the anti-inflammatory effect of Gentiana macrophylla extract. Furthermore, this invention combines Platycladus orientalis leaf extract, Norwegian spruce leaf extract, and the Gentiana macrophylla extract prepared in this invention. The three components synergistically enhance the inhibitory effect of the composition on Propionibacterium acnes, and the effect is even better within a specific dosage ratio range. Simultaneously, the acne-removing composition of this invention can exert acne-removing effects from multiple aspects, including anti-inflammatory and antibacterial properties, and oil control, reducing the area of ​​facial acne, alleviating facial inflammation, and reducing sebum secretion.

[0008] Furthermore, the acne-removing composition comprises the following components in parts by weight: 4-5 parts Gentiana macrophylla extract, 2-3 parts Platycladus orientalis leaf extract, and 0.5-1.5 parts Norway spruce leaf extract.

[0009] Furthermore, the preparation method of the Gentiana macrophylla extract includes the following steps:

[0010] S1. Pulverize the dried Gentiana macrophylla, sieve it, add sterile water, mix well, sterilize, and obtain the fermentation substrate;

[0011] S2. Based on the volume of the fermentation substrate, inoculate with 3%-8% of a mixed inoculum consisting of Kluyveromyces martensii culture and Bacillus pumilus culture to obtain the fermentation broth;

[0012] S3. The fermentation broth is subjected to ultrasonic treatment, sterilization, and filtration. The filtrate is then concentrated and dried to obtain Gentiana macrophylla extract.

[0013] Preferably, in step S2, the volume ratio of Kluyveromyces martensii culture to Bacillus pumilus culture in the mixed bacterial agent is 1:(1-3); the viable cell counts of the Kluyveromyces martensii culture and the Bacillus pumilus culture are the same, both being 10. 6 -10 8 CFU / mL.

[0014] Preferably, the fermentation in step S2 is carried out at 28-40°C for 24-48 hours.

[0015] Preferably, the ultrasonic treatment in step S3 is ultrasonic treatment at 60-70℃ and 200-300W for 30-40 minutes.

[0016] Secondly, the present invention provides the use of the acne-removing composition described in the first aspect in the preparation of cosmetics with acne-removing effects.

[0017] Furthermore, the acne-removing composition accounts for 0.5%-10% of the total mass of the cosmetic.

[0018] Furthermore, the cosmetics include lotions, serums, creams, or masks.

[0019] Thirdly, the present invention provides a cosmetic with acne-removing effects, comprising the acne-removing composition described in the first aspect and excipients acceptable in the cosmetic field.

[0020] Furthermore, the acne-removing composition accounts for 0.5%-10% of the total mass of the cosmetic, preferably 1%-5%.

[0021] Furthermore, the excipients acceptable in the cosmetics field include, but are not limited to, one or more of the following: moisturizers, thickeners, emulsifiers, skin moisturizers, preservatives, and deionized water.

[0022] Preferably, the cosmetic excipients include glycerin, butylene glycol, xanthan gum, 1,2-hexanediol, p-hydroxyphenylethanol, and deionized water.

[0023] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0024] The Gentiana macrophylla extract of this invention is prepared by mixed fermentation of Gentiana macrophylla with a specific strain and a specific ratio of Kluyveromyces martensii and Bacillus pumilus, followed by ultrasonic treatment. The resulting Gentiana macrophylla extract significantly enhances the inhibitory effect on inflammatory factors TNF-α and IL-6, indicating a greatly improved anti-inflammatory effect. This invention combines Platycladus orientalis leaf extract, Norwegian spruce leaf extract, and the Gentiana macrophylla extract prepared in this invention. Testing showed that the three components synergistically enhance the inhibitory effect of the composition on Propionibacterium acnes, with even better results within a specific ratio range. Simultaneously, the acne-removing composition of this invention exerts its acne-removing effect from multiple aspects, including anti-inflammatory and antibacterial properties, and oil control, reducing facial acne area, alleviating facial inflammation, and decreasing sebum secretion. Detailed Implementation

[0025] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0026] The materials used in the following embodiments are sourced from:

[0027] Arborvitae leaf extract: purchased from Shanghai Huiwen Biotechnology Co., Ltd., trade name: Arborvitae leaf extract Hairlux;

[0028] Norway spruce leaf extract: purchased from Shanghai Hongjiu Enterprise Development Co., Ltd., product name: Spruce Hydrosol.

[0029] Gentiana macrophylla is a plant belonging to the Gentianaceae family. Gentiana macrophulla Dried roots of Pall. are commercially available.

[0030] Unless otherwise specified, all other materials and reagents used in the examples are commercially available.

[0031] Example 1

[0032] This embodiment provides a method for preparing Gentiana macrophylla extract 1, including the following steps:

[0033] S1. Pulverize the dried Gentiana macrophylla, pass it through a 100-mesh sieve, add 20 times its weight of sterile water, mix well, sterilize, and obtain the fermentation substrate;

[0034] S2. Based on the volume of the fermentation substrate, inoculate with 5% of a mixed inoculum consisting of Kluyveromyces martensii culture and Bacillus pumilus culture, and ferment at 37°C for 36 hours to obtain the fermentation broth;

[0035] S3. The fermentation broth was ultrasonically treated at 65℃ and 250W for 35 minutes, sterilized, filtered, and the filtrate was concentrated and dried to obtain Gentiana macrophylla extract 1.

[0036] The preparation method of the mixed bacterial agent in step S2 is as follows: Kluyveromyces martensii (CGMCC NO. 13499) and Bacillus pumilus (CGMCC NO: 16914) are activated and cultured separately to obtain a concentration of 1×10⁻⁶ for each. 7 The Kluyveromyces macrocephalae culture and Bacillus pumilus culture were mixed at a volume ratio of 1:2 to obtain a mixed inoculum.

[0037] Example 2

[0038] This embodiment provides a method for preparing Gentiana macrophylla extract 2, wherein the volume ratio of Kluyveromyces martensii culture and Bacillus pumilus culture in the mixed bacterial agent used is 1:1, and the remaining steps and parameters are the same as in Example 1, thereby obtaining Gentiana macrophylla extract 2.

[0039] Example 3

[0040] This embodiment provides a method for preparing Gentiana macrophylla extract 3, wherein the volume ratio of Kluyveromyces martensii culture and Bacillus pumilus culture in the mixed bacterial agent used is 1:3, and the remaining steps and parameters are the same as in Example 1, thereby obtaining Gentiana macrophylla extract 3.

[0041] Example 4

[0042] This embodiment provides a method for preparing Gentiana macrophylla extract 4, wherein the mixed microbial agent used is the same as in Example 1, and the preparation method includes the following steps:

[0043] S1. Pulverize the dried Gentiana macrophylla, pass it through a 100-mesh sieve, add 20 times its weight of sterile water, mix well, sterilize, and obtain the fermentation substrate;

[0044] S2. Based on the volume of the fermentation substrate, inoculate with 8% of the mixed bacterial agent and ferment at 28℃ for 48 hours to obtain the fermentation broth;

[0045] S3. The fermentation broth was ultrasonically treated at 60℃ and 200W for 40 minutes, sterilized, filtered, and the filtrate was concentrated and dried to obtain Gentiana macrophylla extract 4.

[0046] Example 5

[0047] This embodiment provides a method for preparing Gentiana macrophylla extract 5, wherein the mixed microbial agent used is the same as in Example 1, and the preparation method includes the following steps:

[0048] S1. Pulverize the dried Gentiana macrophylla, pass it through a 100-mesh sieve, add 20 times its weight of sterile water, mix well, sterilize, and obtain the fermentation substrate;

[0049] S2. Based on the volume of the fermentation substrate, inoculate with 3% mixed bacterial agent and ferment at 40℃ for 24 hours to obtain the fermentation broth;

[0050] S3. The fermentation broth was ultrasonically treated at 70℃ and 300W for 30 minutes, sterilized, filtered, and the filtrate was concentrated and dried to obtain Gentiana macrophylla extract 5.

[0051] Comparative Example 1

[0052] The only difference between this comparative example and Example 1 is that an equal volume of Max Kluyveromyces broth was used instead of the mixed inoculum for fermentation. All other steps and parameters were the same as in Example 1, and Gentiana macrophylla extract 6 was obtained.

[0053] Comparative Example 2

[0054] The only difference between this comparative example and Example 1 is that an equal volume of Bacillus simulans bacterial solution was used instead of the mixed bacterial agent for fermentation. All other steps and parameters were the same as in Example 1, and Gentiana macrophylla extract 7 was obtained.

[0055] Comparative Example 3

[0056] The only difference between this comparative example and Example 1 is that the Saccharomyces cerevisiae with accession number CICC NO.1596 was used instead of the Kluyveromyces martensii used in Example 1; the remaining steps and parameters were the same as in Example 1, and Gentiana macrophylla extract 8 was obtained.

[0057] Comparative Example 4

[0058] The only difference between this comparative example and Example 1 is that the *Kluyveromyces martensii* strain with accession number GDMCC NO: 64440 was used instead of the *Kluyveromyces martensii* strain used in Example 1. All other steps and parameters were the same as in Example 1, and *Gentiana macrophylla* extract 9 was obtained.

[0059] Comparative Example 5

[0060] The only difference between this comparative example and Example 1 is that Bacillus pumilus with accession number CGMCC 1.7925 is used instead of Bacillus pumilus used in Example 1. All other steps and parameters are the same as in Example 1, and Gentiana macrophylla extract 10 is obtained.

[0061] Comparative Example 6

[0062] The only difference between this comparative example and Example 1 is that the volume ratio of Kluyveromyces martensii culture and Bacillus pumilus culture in the mixed bacterial agent in step S2 is 2:1. All other steps and parameters are the same as in Example 1, and Gentiana macrophylla extract 11 is obtained.

[0063] Comparative Example 7

[0064] This comparative example provides a method for preparing Gentiana macrophylla extract 12, which includes the following steps: pulverizing dried Gentiana macrophylla, passing it through a 100-mesh sieve, adding 20 times its weight of sterile water and mixing well, ultrasonically treating it at 65℃ and 250W for 35 minutes, sterilizing it, filtering it, concentrating and drying the filtrate to obtain Gentiana macrophylla extract 12.

[0065] Test Example 1

[0066] Gentiana macrophylla extracts 1-12 prepared in Examples 1-5 and Comparative Examples 1-7 were used as test samples to determine their anti-inflammatory efficacy. The specific experimental steps are as follows:

[0067] (1) Inoculation: The cell line used was HaCaT cells. The cells were seeded into 24-well plates and cultured at 37°C with 5% CO2.

[0068] (2) LPS induction: After culturing for 24 h, the culture medium in the plate was aspirated and washed once with PBS. DEME cell culture medium was added to the blank control group, and DEME cell culture medium containing 1 μg / mL LPS was added to the negative control group and the sample group. The plates were cultured at 37℃ and 5% CO2 for 24 h. Each treatment group had 3 replicates.

[0069] (3) After induction, the culture medium was replaced: DEME cell culture medium was added to both the blank control group and the negative control group, and an equal amount of DEME cell culture medium containing 1 mg / mL of test sample was added to the sample group. The samples were cultured at 37°C and 5% CO2 for 24 h.

[0070] (4) Detection of inflammatory factor content: Take the cell supernatant from each well and detect the content of cellular inflammatory factors TNF-α and IL-6 according to the ELISA kit instructions, and calculate the inflammatory factor inhibition rate; the calculation formula is shown below:

[0071] Inflammatory factor inhibition rate (%) = (average concentration of negative control group - average concentration of sample group) / average concentration of negative control group × 100%.

[0072] The results are shown in Table 1. Compared with Gentiana macrophylla extract 12, which was not fermented but only sonicated, Gentiana macrophylla extracts 1-11, which were fermented and then sonicated, showed stronger inhibitory effects on TNF-α and IL-6, with Gentiana macrophylla extract 1 exhibiting the strongest inhibitory effect on both TNF-α and IL-6. Compared with Gentiana macrophylla extract 1, Gentiana macrophylla extracts 6-7 were fermented using only a single strain, while Gentiana macrophylla extracts 8-10 replaced the fermentation strain. The inhibition rates of TNF-α and IL-6 of Gentiana macrophylla extract 1 were significantly higher than those of Gentiana macrophylla extracts 6-10. Therefore, it can be seen that fermenting Gentiana macrophylla with specific strains of Kluyveromyces martensii and Bacillus pumilus can synergistically enhance the anti-inflammatory ability of Gentiana macrophylla extract. Compared with Gentiana macrophylla extracts 1-3, the volume ratio of Kluyveromyces martensii and Bacillus pumilus in the mixed bacterial agent of Gentiana macrophylla extract 11 is not within the range limited by this invention, and the inhibitory effect of Gentiana macrophylla extract 11 on TNF-α and IL-6 is significantly lower than that of Gentiana macrophylla extracts 1-3, indicating that the ratio of Kluyveromyces martensii and Bacillus pumilus in the fermentation agent has an important influence on the anti-inflammatory ability of Gentiana macrophylla extract.

[0073] Table 1. Inhibitory effects of Gentiana macrophylla extract on inflammatory factors TNF-α and IL-6

[0074]

[0075] Application Examples 1-5

[0076] Based on the results of Test Example 1, it can be seen that Gentiana macrophylla extract 1 has the best anti-inflammatory effect. Therefore, in this application example, Gentiana macrophylla is used to form an acne-removing composition with other plant extracts, and then further prepared into an essence for application effect testing.

[0077] Specifically, Gentiana macrophylla extract 1 was compounded with Platycladus orientalis leaf extract and Norwegian spruce leaf extract to obtain different acne-removing compositions. These acne-removing compositions were prepared according to the formulas in Table 2, resulting in acne-removing compositions 1-5. Then, the acne-removing compositions 1-5 prepared above were used to prepare serums, resulting in serums 1-5.

[0078] Table 2. Components and Dosage of Acne Treatment Composition

[0079]

[0080] The serum comprises the following components by weight percentage: 3% glycerin, 4.5% butylene glycol, 0.2% xanthan gum, 0.5% 1,2-hexanediol, 0.5% p-hydroxyphenylethanol, 3% acne-fighting composition, and the balance being deionized water.

[0081] The preparation method of the essence is as follows: Glycerin, butylene glycol, xanthan gum and deionized water are mixed evenly according to the formula. The resulting mixture is kept at 85°C for 30 minutes. When the temperature drops to 60°C, 1,2-ethylene glycol and p-hydroxyphenylethanol are added and stirred evenly. The mixture is then cooled to 45°C, and the acne-removing composition is added and stirred evenly to obtain the essence.

[0082] Comparative Application Example 1

[0083] This comparative application example provides an acne-removing composition 6, which differs from composition 3 in that it does not contain Gentiana macrophylla extract 1. Instead, the missing amount is made up by Platycladus orientalis leaf extract and Norwegian spruce leaf extract in a weight ratio of 2.5:1, thus obtaining acne-removing composition 6.

[0084] This comparative application example also provides a serum 6, which differs from serum 3 only in that an equal amount of acne-removing composition 3 is replaced with acne-removing composition 6.

[0085] Comparative Application Example 2

[0086] This comparative application example provides an acne-removing composition 7, which differs from acne-removing composition 3 in that it does not contain Platycladus orientalis leaf extract. Instead, the missing amount is made up by Gentiana macrophylla extract 1 and Norwegian spruce leaf extract in a weight ratio of 4.5:1, resulting in acne-removing composition 7.

[0087] This comparative application example also provides a serum 7, which differs from serum 3 only in that an equal amount of acne-removing composition 3 is replaced with acne-removing composition 7.

[0088] Comparative Application Example 3

[0089] This comparative application example provides an acne treatment composition 8, which differs from acne treatment composition 3 in that it does not contain Norway spruce leaf extract. Instead, the missing amount is made up by Gentiana macrophylla extract 1 and Platycladus orientalis leaf extract in a weight ratio of 4.5:2.5, thus obtaining acne treatment composition 8.

[0090] This comparative application example also provides a serum 8, which differs from serum 3 only in that an equal amount of acne-removing composition 3 is replaced with acne-removing composition 8.

[0091] Comparative Application Example 4

[0092] This comparative application example provides an acne treatment composition 9, which contains 2.5 parts of Gentiana macrophylla extract, 4.5 parts of Platycladus orientalis leaf extract, and 1 part of Norwegian spruce leaf extract.

[0093] This comparative application example also provides a serum 9, which differs from serum 3 only in that an equal amount of acne-removing composition 3 is replaced with acne-removing composition 9.

[0094] Comparative Application Example 5

[0095] This comparative application example provides an acne treatment composition 10, which contains 1 part Gentiana macrophylla extract, 2.5 parts Platycladus orientalis leaf extract and 4.5 parts Norway spruce leaf extract.

[0096] This comparative application example also provides a serum 10, which differs from serum 3 only in that an equal amount of acne-removing composition 3 is replaced with acne-removing composition 10.

[0097] Test Example 2

[0098] This test case evaluates the acne-removing efficacy of acne-removing compositions 1-10 by testing their inhibitory effect on Propionibacterium acnes.

[0099] Test method:

[0100] Acne-removing compositions 1-10 were prepared as test samples with a mass concentration of 5%. The antibacterial effect was tested using the inhibition zone method with agar medium. The test strain was *Propionibacterium acnes*, and the bacterial suspension was prepared to a concentration of 2 × 10⁻⁶. 6 A bacterial suspension of CFU / mL was used for testing.

[0101] Antibacterial effect determination: 200 μL of Propionibacterium acnes suspension was evenly spread onto an agar plate, and then the plate was placed in a sterile operating table to dry. Sterilized Oxford cups were placed in the center of the plate, ensuring complete contact with the culture medium. The test sample was quantitatively added to the Oxford cup, with three replicates for each sample. The plates were anaerobically incubated at 37℃ for 2 days, and the results were recorded as the diameter of the inhibition zone (mm), and the mean was calculated. Evaluation criteria for antibacterial effect: Inhibition zone diameter > 20 mm was considered significant antibacterial; inhibition zone diameter 10-20 mm was considered moderate antibacterial; inhibition zone diameter < 10 mm was considered no antibacterial.

[0102] Propionibacterium acnes is the main pathogen causing acne. Therefore, the inhibitory effect of acne-removing compositions on Propionibacterium acnes can reflect their acne-removing efficacy. Table 3 shows that the inhibition zone diameter of acne-removing compositions 1-5 against Propionibacterium acnes reaches 20.58-25.46 mm, indicating a significant inhibitory effect and potential acne-removing efficacy in practical applications. Comparing the inhibition zone diameter of acne-removing compositions 6-10 with that of Propionibacterium acnes, acne-removing composition 3 shows an increased inhibition zone diameter of 9.63-15.08 mm. The only difference between acne-removing compositions 6-10 and acne-removing composition 3 is the absence of one extract component or a different weight ratio of the components, indicating that the three components in the acne-removing compositions interact to jointly enhance the inhibitory effect of the composition on Propionibacterium acnes.

[0103] Table 3. Results of inhibition zone diameter of the composition against Propionibacterium acnes

[0104]

[0105] Test Example 3

[0106] 1. Skin patch test

[0107] The safety of the serums prepared in the application examples and comparative application examples (1-10) on human skin was verified. In accordance with the "Cosmetic Safety Technical Specifications" (2015) for human skin patch testing, 100 volunteers aged 18-50 years were recruited. The test environment was a temperature of (21±1)℃ and a humidity of (50±10)%. Volunteers washed their forearms with water. After 5-10 minutes, once the moisture had evaporated, 0.025 mL of each sample was applied to each chamber of a 10-well patch applicator. The inner sides of both forearms of the volunteers were selected, avoiding the skin at the joints of the front and back of the hands. Each arm was covered with a patch applicator and marked accordingly. The patch application lasted for 24 hours. After removing the patch applicator and wiping away any remaining sample, skin reactions were observed at 0.5h, 24h, and 48h. The test results were recorded according to the adverse skin reaction grading standards in Table 4.

[0108] Table 4. Severity of Adverse Skin Reactions

[0109]

[0110] After testing, the essences 1-10 of this invention showed negative reactions after human patch testing, indicating that they are safe and non-irritating to human skin.

[0111] 2. Acne treatment effect test

[0112] The serums 1-10 prepared in the application examples and comparative application examples were used as samples to test their acne-removing effects. The test method is as follows:

[0113] We recruited 100 volunteers with grade 1-2 acne on their faces, randomly divided into groups of 10, with each group receiving one sample. The method of use was as follows: after cleansing the face, apply 1mL of the serum and massage it in until absorbed, twice a day, morning and evening, for 14 consecutive days.

[0114] The facial skin condition of volunteers was tested on days 0 and 14. On the day of the visit, volunteers washed their faces with water without applying any products and sat quietly for 20 minutes in an air-conditioned room with a temperature of 21±1℃ and humidity of 50±10%. The size of acne lesions and the a-value (a-value, representing the erythema content of the skin in the dermis, indicating the state of skin inflammation) were measured using a VISA instrument. Facial oil content was measured using a skin oil test probe. Tests were performed three times before and after the experiment, and the average values ​​were taken. The improvement rate was calculated using the following formula:

[0115] Improvement rate (%) = [(X mean before use - X mean after use) / X mean before use] × 100%.

[0116] This test case reflects the acne-removing effect of the acne-removing composition by measuring the acne area, skin a value, and oil content on the volunteers' faces after 14 days of use of the serum. The results are averaged and are shown in Table 5. It can be seen that serums 1-5 have a better improvement effect on the acne area, skin a value, and facial oil content of volunteers' faces compared with serums 6-10. The only difference between serums 6-10 and serum 3 is that the acne-removing composition of the serums lacks any one of Gentiana macrophylla extract, Platycladus orientalis leaf extract, and Norwegian spruce leaf extract, or the proportion of the three components in the acne-removing composition is different. This indicates that the components in the acne-removing composition and their proportions are important factors affecting the acne-removing effect of the composition.

[0117] Table 5 Results of facial skin condition of volunteers after 14 days of use.

[0118]

[0119] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. An acne-removing composition, characterized in that, It is composed of 3-6 parts Gentiana macrophylla extract, 1-4 parts Platycladus orientalis leaf extract and 0.5-2 parts Norwegian spruce leaf extract; the Gentiana macrophylla extract is prepared by using Gentiana macrophylla as raw material, through mixed fermentation of Kluyveromyces martensii and Bacillus pumilus, followed by ultrasonic treatment. The preparation method of the Gentiana macrophylla extract includes the following steps: S1. Pulverize the dried Gentiana macrophylla, sieve it, add sterile water, mix well, sterilize, and obtain the fermentation substrate; S2. Based on the volume of the fermentation substrate, inoculate 3%-8% of a mixed inoculum consisting of Kluyveromyces martensii and Bacillus pumilus inoculum, and ferment at 28-40℃ for 24-48 hours to obtain the fermentation broth; the volume ratio of Kluyveromyces martensii to Bacillus pumilus inoculum in the mixed inoculum is 1:(1-3); the viable cell counts of the Kluyveromyces martensii and Bacillus pumilus inoculum are the same, both being 10. 6 -10 8 CFU / mL; S3. The fermentation broth is subjected to ultrasonic treatment, sterilization, and filtration. The filtrate is then concentrated and dried to obtain Gentiana macrophylla extract. The preservation number of *Kluyveromyces martensii* is CGMCC NO.13499, and the preservation number of *Bacillus pumilus* is CGMCC NO:16914.

2. The acne-removing composition as described in claim 1, characterized in that, It consists of 4-5 parts Gentiana macrophylla extract, 2-3 parts Platycladus orientalis leaf extract and 0.5-1.5 parts Norway spruce leaf extract.

3. The acne-removing composition as described in claim 1, characterized in that, The ultrasonic treatment in step S3 is ultrasonic treatment at 60-70℃ and 200-300W for 30-40 minutes.

4. The use of the acne-removing composition according to any one of claims 1-3 in the preparation of cosmetics with acne-removing effects.

5. The application as described in claim 4, characterized in that, The acne-removing composition accounts for 0.5%-10% of the total mass of the cosmetic.

6. The application as described in claim 4, characterized in that, The cosmetics include lotions, serums, creams, or masks.

7. A cosmetic product with acne-removing effects, characterized in that, It contains the acne-removing composition according to any one of claims 1-3 and excipients acceptable in the cosmetic field.

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