A multi-pass high-efficiency oil-control and acne-removing composition and its application in cosmetics

Abstract

This invention relates to the field of cosmetics, and more particularly to a multi-pathway, highly effective oil-controlling and acne-reducing composition and its application in cosmetics. The oil-controlling and acne-reducing composition of this invention comprises the following components in parts by weight: 1-5 parts capryloyl salicylic acid, 0.5-2 parts magnolol, and 0.5-10 parts white willow bark extract. The synergistic effect of capryloyl salicylic acid, magnolol, and white willow bark extract, when combined, exhibits significant oil-controlling and acne-reducing effects, and can be applied to cosmetics of various formulations. Furthermore, this invention also found that the oil-controlling and acne-reducing effects of white willow bark extract can be further enhanced after enzymatic modification.

Description

Technical Field

[0001] This invention relates to the field of cosmetics, and more particularly to a multi-channel, highly effective oil-controlling and acne-removing composition and its application in cosmetics. Background Technology

[0002] Excessive facial oil secretion is essentially a condition called seborrhea. Seborrhea is mainly caused by overactive sebaceous glands on the face, leading to excessive sebum production. Areas with large pores are more prone to producing excess oil, resulting in oily skin. The overactivity of sebaceous glands is primarily due to stimulation by dihydrotestosterone (DHT), a component of androgens. The formation of androgens can be caused by genetic factors, as well as insufficient sleep, frequent consumption of spicy and stimulating foods, leading to excessive androgen secretion, especially DHT, which stimulates the sebaceous glands to produce excessive sebum. Excessive sebum secretion can then lead to other skin conditions, commonly manifesting as open comedones (blackheads) and potentially worsening into acne.

[0003] Surveys indicate that 75-90% of teenagers and young adults suffer from acne, with nearly 10-20% of people aged 25-35 still experiencing acne, and 5% of men and women still having acne until the age of 45. Acne causes significant distress and can lead to various complications.

[0004] Many oil-control and acne-fighting products contain strong cleansing ingredients such as alcohol and benzoic acid. While these can effectively remove oil, long-term use may lead to dry and dehydrated skin. Dry skin produces more oil to protect itself, potentially creating a vicious cycle where the more you try to control oil, the oilier it becomes. Many oil-control and acne-fighting products only temporarily relieve symptoms and do not address the root cause of the problem. Once you stop using them, acne and oiliness may reappear, offering only temporary relief. Summary of the Invention

[0005] The purpose of this invention is to overcome the shortcomings of the existing technology and provide an oil-controlling and acne-removing composition with significant oil-controlling and acne-removing effects, which can be applied to cosmetics.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0007] In a first aspect, the present invention provides an oil-controlling and acne-removing composition comprising the following components in parts by weight: 1-5 parts of capryloyl salicylic acid, 0.5-2 parts of magnolol, and 0.5-10 parts of white willow bark extract.

[0008] Capryloyl salicylic acid has exfoliating, anti-inflammatory, and sebum-regulating effects. It can penetrate deep into pores to help remove dead skin cells, reduce the formation of acne and blackheads, reduce redness and inflammation, and is beneficial for acne-prone skin. It also controls excessive sebum secretion and prevents clogged pores. Magnolia officinalis has antibacterial, antiviral, antioxidant, anti-inflammatory, and soothing effects. It has a strong antibacterial effect, helping to inhibit the growth of microorganisms that cause skin infections; it can scavenge free radicals, protecting the skin from environmental stressors; and it relieves skin irritation and discomfort. White willow bark extract is a natural source of salicylic acid, containing abundant salicylic acid derivatives, and has anti-inflammatory and antipyretic effects similar to aspirin; it also has anti-inflammatory and analgesic effects, significantly relieving skin inflammation and pain. This invention combines these three components for synergistic effects, comprehensively leveraging their efficacy to achieve significant oil control and acne-reducing effects, effectively improving or treating skin problems.

[0009] Preferably, the oil-controlling and acne-removing composition comprises the following components in parts by weight: 2 parts capryloyl salicylic acid, 1 part magnolol, and 1.2 parts white willow bark extract.

[0010] Preferably, the white willow bark extract comprises unmodified white willow bark extract and / or enzyme-modified white willow bark extract; the enzyme comprises at least one of β-glucosidase, β-glucanase, and xylanase.

[0011] White willow is a plant species recognized in the field as having potential for edible, medicinal, and cosmetic use. The properties of its bark extract have been extensively studied. In order to improve the repair effect of white willow bark extract, this invention performs ultrasonic extraction and enzyme modification treatment on it. It was found that enzyme modification has a positive impact on the efficacy of white willow bark extract.

[0012] More preferably, the white willow bark extract is obtained by modification with β-glucosidase.

[0013] β-glucosidase is an important component of cellulase, present in many plants, and is involved in the relaxation or strengthening of cell walls during plant cell growth and development. It is also associated with signal recognition and the production of certain signaling molecules in plant cells. This invention uses β-glucosidase to modify white willow bark extract. Under the action of this enzyme, many precursor substances in white willow bark, such as salicylates and active ingredients in bonded volatile substances, can be better released, thus allowing the white willow bark extract to exert its efficacy more effectively.

[0014] Preferably, the preparation method of the unmodified white willow bark extract includes the following steps: adding water to white willow bark to extract white willow bark aqueous extract, concentrating, adding heterobranching acid synthase as an aid, stirring and adding ethanol, sealing and standing, filtering to recover ethanol, drying, and the resulting powder is the white willow bark extract.

[0015] Preferably, the preparation method of the unmodified white willow bark extract specifically includes the following steps:

[0016] Add five times the mass of deionized water to the pulverized white willow bark for extraction, controlling the temperature between 50-60℃ and stirring for 10-20 minutes; let stand for 180-240 minutes, and perform ultrasonic treatment for 10 minutes every 20 minutes during this period to help separate the effective components, obtaining a white willow bark aqueous extract. Concentrate to 1:(1-2)(mL:g), add 0.2% isobranching acid synthase (ICS1) to assist, and after cooling, slowly add ethanol while stirring to reach the specified alcohol content (above 75%). Refrigerate in a sealed container for 24-48 hours, filter, recover the ethanol from the filtrate to obtain a purified liquid, dry, and the resulting powder is the white willow bark extract.

[0017] Preferably, the preparation method of the enzyme-modified white willow bark extract includes the following steps: (1) adding water to white willow bark to extract white willow bark aqueous extract, concentrating, adding heterobranching acid synthase as an aid, stirring and adding ethanol, sealing and standing, filtering to recover ethanol, drying to obtain powder, and adding water to prepare an aqueous solution; (2) adding enzyme modification treatment to the aqueous solution obtained in step (1), concentrating, and freeze-drying to obtain the white willow bark extract.

[0018] This invention uses white willow, widely cultivated in Xinjiang, China, as raw material. The water extract of white willow (SALIXALBA) bark, prepared by the applicant, is obtained using a water extraction and alcohol precipitation method. 0.2% isobranching acid synthase (ICSI) is added as an aid during extraction. Microwave drying technology is used, and the dried water extract is a brown or grayish-white fine powder. The salicylic acid content in the white willow bark water extract powder is >18%.

[0019] Preferably, the concentration of the aqueous solution is 8-12% w / v.

[0020] Dissolve 8-12g of the obtained powder in water to prepare an aqueous solution with a concentration of 8-12% w / v.

[0021] Preferably, in step (2), the concentration of the enzyme is 500-800 U / mL.

[0022] More preferably, in step (2), the concentration of the enzyme is 700 U / mL.

[0023] Preferably, in step (2), the enzyme treatment conditions are: adjusting the pH to 5.0-6.0 and treating at 50-60℃ for 1-3 hours.

[0024] This invention has revealed that within a certain pH range, the rate of enzymatic reaction in plant enzymes increases with increasing pH; however, when the pH rises to a certain limit, the rate of enzymatic reaction not only stops increasing but actually decreases with further increases in pH. This invention has found that enzyme activity is optimal at a pH of 5.0-6.0, which represents the optimal pH range for enzyme reactions.

[0025] More preferably, in step (2), the enzyme treatment conditions are: adjusting the pH to 5.5 and treating at 55°C for 2 hours.

[0026] Preferably, in step (2), the enzyme is inactivated by treating it at 90°C for 3-6 minutes after treatment.

[0027] Secondly, the present invention provides the application of the above-mentioned oil-controlling and acne-removing composition in cosmetics.

[0028] Thirdly, the present invention provides a cosmetic product comprising the above-described oil-controlling and acne-removing composition.

[0029] Preferably, the cosmetic also includes excipients required for daily chemical products.

[0030] Preferably, the excipients include emulsifiers, skin conditioning agents, moisturizers, antioxidants, thickeners, emollients, and pH adjusters.

[0031] Preferably, the emulsifier comprises PEG-80 sorbitan lauryl ester; the skin conditioning agent comprises Centella Asiatica extract; the moisturizer comprises at least one of butylene glycol, 1,2-hexanediol, and glycerin; the antioxidant comprises p-hydroxyacetophenone; the thickener comprises at least one of acrylate / C10-30 alkanol acrylate crosspolymer, sodium polyacrylamide dimethyl taurate, and Caesalpinia spicosa gum; the emollient comprises polydimethylsiloxane and caprylic / capric triglyceride; and the pH adjuster comprises sodium hydroxide.

[0032] The beneficial effects of this invention are as follows:

[0033] This invention combines capryloyl salicylic acid, magnolol, and white willow bark extract. The three components work synergistically to achieve significant oil-controlling and acne-reducing effects, making it applicable to various cosmetic formulations. Furthermore, this invention also reveals that enzymatic modification of the white willow bark extract further enhances its oil-controlling and acne-reducing effects. Attached Figure Description

[0034] Figure 1 Effects of unmodified / enzyme-modified white willow bark extracts on the survival rate of SZ95 cells.

[0035] Figure 2Effects of unmodified / enzyme-modified white willow bark extracts on the antibacterial rate of Staphylococcus aureus.

[0036] Figure 3 Effects of unmodified / enzyme-modified white willow bark extracts on the antibacterial rate of Propionibacterium acnes. Detailed Implementation

[0037] 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.

[0038] Unless otherwise specified, all raw materials used in this invention can be purchased from the market.

[0039] Heterobrominated acid synthase (ICS1), β-glucosidase, β-glucanase, and xylanase were purchased from manufacturers such as Shanghai Yuanye and Wuhan Huaxiang Kejie, and all were food grade.

[0040] The raw materials used in the cosmetics in this invention are cosmetic grade.

[0041] The bark extract of white willow (SALIX ALBA) used white willow, which is widely cultivated in Xinjiang, China. The extraction method was water extraction and alcohol precipitation, with 0.2% isobranching acid synthase (ICS1) added as an aid. The salicylic acid content of the extract was >18%. Capryloyl salicylic acid was purchased from Shanghai Bilai Chemical Technology Co., Ltd., and magnolol was purchased from Hunan Heguang Biotechnology Co., Ltd.

[0042] Example 1:

[0043] An embodiment of the oil-controlling and acne-removing composition of the present invention; the oil-controlling and acne-removing composition comprises the following components in parts by weight: 2 parts capryloyl salicylic acid, 1 part magnolol, and 1.2 parts white willow bark extract.

[0044] The white willow bark extract was obtained by water extraction and alcohol precipitation.

[0045] The preparation method of the white willow bark extract (using water extraction and alcohol precipitation with enzyme assistance) includes the following steps:

[0046] Add five times the mass of deionized water to the pulverized white willow bark for extraction, controlling the temperature between 50-60℃ and stirring for 10-20 minutes; let stand for 180-240 minutes, and during this period, perform ultrasonic treatment for 10 minutes every 20 minutes to help separate the effective components, to obtain the white willow bark aqueous extract, concentrate to 1:(1-2)(mL:g), add 0.2% isobranching acid synthase (ICS1) to assist, cool, and slowly add ethanol while stirring to reach the specified alcohol content (above 75%), seal and refrigerate for 24-48 hours, filter, recover the ethanol from the filtrate to obtain the purified liquid, and dry the resulting powder to obtain the white willow bark extract.

[0047] Example 2:

[0048] An embodiment of the oil-controlling and acne-removing composition of the present invention; the oil-controlling and acne-removing composition comprises the following components in parts by weight: 2 parts capryloyl salicylic acid, 1 part magnolol, and 1.2 parts white willow bark extract.

[0049] The white willow bark extract was obtained by modification with β-glucosidase.

[0050] The preparation method of the white willow bark extract includes the following steps: (1) Add five times the mass of deionized water to the pulverized white willow bark for extraction, control the temperature between 50-60℃, stir for 10-20 min; let stand for 180-240 min, and perform ultrasonic treatment for 10 min every 20 min to help separate the effective components, obtain the white willow bark aqueous extract, concentrate it to 1:(1-2)(mL:g), and add 0.2% isobranching acid synthase (ICS1). (1) After cooling, slowly add ethanol while stirring to reach the specified alcohol content (above 75%), seal and refrigerate for 24-48 hours, filter, recover ethanol from the filtrate to obtain a purified liquid, dry to obtain powder, add water to prepare a 10% w / v aqueous solution; (2) Add 700 U / mL β-glucosidase to the aqueous solution in step (1), adjust the pH to 5.5, treat at 55℃ for 2 hours, then treat at 90℃ for 5 minutes to inactivate the enzyme, concentrate, freeze dry to obtain the white willow bark extract.

[0051] Example 3:

[0052] An embodiment of the oil-controlling and acne-removing composition of the present invention; the oil-controlling and acne-removing composition comprises the following components in parts by weight: 2 parts capryloyl salicylic acid, 1 part magnolol, and 1.2 parts white willow bark extract.

[0053] The white willow bark extract was obtained by modification with β-glucanase.

[0054] The preparation method of the white willow bark extract includes the following steps: (1) Add five times the mass of deionized water to the pulverized white willow bark for extraction, control the temperature between 50-60℃, stir for 10-20 min; let stand for 180-240 min, and perform ultrasonic treatment for 10 min every 20 min to help separate the effective components, obtain the white willow bark aqueous extract, concentrate it to 1:(1-2)(mL:g), and add 0.2% isobranching acid synthase (ICS). 1) After cooling, slowly add ethanol while stirring to reach the specified alcohol content (above 75%), seal and refrigerate for 24-48 hours, filter, recover ethanol from the filtrate to obtain a purified liquid, dry to obtain powder, add water to prepare a 10% w / v aqueous solution; (2) Add 700 U / mL β-glucanase to the aqueous solution in step (1), adjust the pH to 5.5, treat at 55°C for 2 hours, then treat at 90°C for 5 minutes to inactivate the enzyme, concentrate, freeze dry to obtain the white willow bark extract.

[0055] Example 4:

[0056] An embodiment of the oil-controlling and acne-removing composition of the present invention; the oil-controlling and acne-removing composition comprises the following components in parts by weight: 2 parts capryloyl salicylic acid, 1 part magnolol, and 1.2 parts white willow bark extract.

[0057] The white willow bark extract was obtained by modification with xylanase.

[0058] The preparation method of the white willow bark extract includes the following steps: (1) Add five times the mass of deionized water to the pulverized white willow bark for extraction, control the temperature between 50-60℃, stir for 10-20 min; let stand for 180-240 min, and perform ultrasonic treatment for 10 min every 20 min to help separate the effective components, obtain the white willow bark aqueous extract, concentrate it to 1:(1-2)(mL:g), and add 0.2% isobranching acid synthase (IC). S1) After cooling, slowly add ethanol while stirring to reach the specified alcohol content (above 75%), seal and refrigerate for 24-48 hours, filter, recover ethanol from the filtrate to obtain a purified liquid, dry to obtain powder, add water to prepare a 10% w / v aqueous solution; (2) Add 700 U / mL xylanase to the aqueous solution in step (1), adjust the pH to 5.5, treat at 55°C for 2 hours, then treat at 90°C for 5 minutes to inactivate the enzyme, concentrate, freeze dry to obtain the white willow bark extract.

[0059] Example 5:

[0060] An embodiment of the oil-controlling and acne-removing composition of the present invention; the oil-controlling and acne-removing composition comprises the following components in parts by weight: 2 parts capryloyl salicylic acid, 1 part magnolol, and 1.2 parts white willow bark extract.

[0061] The white willow bark extract is a commercially available product purchased from Bozhou Xier Traditional Chinese Medicine Co., Ltd., and was prepared using only the water extraction and alcohol precipitation method.

[0062] Example 6:

[0063] An embodiment of the oil-controlling and acne-removing composition of the present invention; the oil-controlling and acne-removing composition comprises the following components in parts by weight: 1 part capryloyl salicylic acid, 0.5 parts magnolol, and 10 parts white willow bark extract.

[0064] The white willow bark extract was obtained by modification with β-glucosidase.

[0065] The preparation of the white willow bark extract is the same as in Example 2.

[0066] Example 7:

[0067] An embodiment of the oil-controlling and acne-removing composition of the present invention; the oil-controlling and acne-removing composition comprises the following components in parts by weight: 5 parts capryloyl salicylic acid, 2 parts magnolol, and 0.5 parts white willow bark extract.

[0068] The white willow bark extract was obtained by modification with β-glucosidase.

[0069] The preparation of the white willow bark extract is the same as in Example 2.

[0070] Comparative Example 1:

[0071] A comparative example of the oil-controlling and acne-removing composition of the present invention; the oil-controlling and acne-removing composition comprises the following components in parts by weight: 6 parts capryloyl salicylic acid, 3 parts magnolol, and 12 parts white willow bark extract.

[0072] The white willow bark extract was obtained by modification with β-glucosidase.

[0073] The preparation of the white willow bark extract is the same as in Example 2.

[0074] Comparative Example 2:

[0075] This invention provides a comparative example of the oil-controlling and acne-removing composition. The only difference between this comparative example and Example 2 is that this comparative example does not contain magnolol, but instead uses an equal part by weight of water.

[0076] Comparative Example 3:

[0077] This invention provides a comparative example of the oil-controlling and acne-removing composition. The only difference between this comparative example and Example 2 is that this comparative example does not contain white willow bark extract, but instead uses an equal weight of water.

[0078] Comparative Example 4:

[0079] This invention provides a comparative example of the oil-controlling and acne-removing composition. The only difference between this comparative example and Example 2 is that this comparative example does not add capryloyl salicylic acid, but instead uses an equal weight of water.

[0080] Application Example 1:

[0081] An application embodiment of the cosmetic described in this invention; the cosmetic is specifically a serum.

[0082] The cosmetic comprises the following components by weight percentage: 0.5% acrylate / C10-30 alkanol acrylate crosspolymer, 0.4% sodium polyacrylamide dimethyl taurate, 0.2% Caesalpinia spironosa gum, 9% glycerin, 5% butylene glycol, 0.5% polydimethylsiloxane, 0.8% caprylic / capric triglyceride, 0.1% sodium hydroxide, 0.45% p-hydroxyacetophenone, 0.5% 1,2-hexanediol, 10% pre-formulated oil-controlling and acne-reducing composition, and the balance being water.

[0083] The pre-formulated oil-controlling and acne-removing composition comprises the following components by weight percentage: 4.2% of the oil-controlling and acne-removing composition of Example 1, 1.5% of PEG-80 dehydrated sorbitan lauryl ester, 1.2% of Centella Asiatica extract, 5% of butylene glycol, 0.45% of p-hydroxyacetophenone, 5% of 1,2-hexanediol, 10% of glycerin, and the balance being water.

[0084] The preparation method of the oil-controlling and acne-removing cosmetic includes the following steps:

[0085] (1) Preparation of the pre-form of the oil-controlling and acne-removing composition: ① The oil-controlling and acne-removing composition, PEG-80 dehydrated sorbitan lauryl ester, and butylene glycol are mixed and heated until completely dissolved; ② Centella asiatica extract, p-hydroxyacetophenone, 1,2-hexanediol, glycerin, and water are added, and the mixture is kept at 80-85℃ and stirred to homogenize and disperse. After stirring and returning to room temperature, the pre-form of the oil-controlling and acne-removing composition is obtained.

[0086] (2) Preparation process of the essence: ① Mix and stir the acrylic (ester) cross-linked polymer / C10-30 alkanol acrylate, sodium polyacrylamide dimethyl taurate, Caesalpinia spironosa gum, and glycerin to obtain mixture A; ② Add butanediol, p-hydroxyacetophenone, 1,2-hexanediol, and water to mixture A, heat to 80-85℃, and homogenize and stir to obtain mixture B; ③ Add polydimethylsiloxane and caprylic / capric triglyceride to mixture B, homogenize and emulsify, and stir and cool to obtain mixture C; ④ Cool to 50-55℃, add sodium hydroxide to mixture C, stir and disperse evenly, and continue stirring and cooling; cool to 30-40℃, add the pre-formed phase of the oil-controlling and acne-removing composition obtained in step (1), stir and disperse evenly, and discharge to obtain the oil-controlling and acne-removing cosmetic (essence).

[0087] Application Example 2:

[0088] An application embodiment of the cosmetic described in this invention; the cosmetic is specifically a serum.

[0089] The only difference between this application example and application example 1 is that the oil-controlling and acne-removing composition of example 1 is replaced with an equal amount of the oil-controlling and acne-removing composition of example 2; all other components and contents are the same as in application example 1. Application example 3:

[0090] An application embodiment of the cosmetic described in this invention; the cosmetic is specifically a serum.

[0091] The only difference between this application example and application example 1 is that the oil-controlling and acne-removing composition of example 1 is replaced with an equal amount of the oil-controlling and acne-removing composition of example 3; all other components and contents are the same as in application example 1. Application example 4:

[0092] An application embodiment of the cosmetic described in this invention; the cosmetic is specifically a serum.

[0093] The only difference between this application example and application example 1 is that the oil-controlling and acne-removing composition of example 1 is replaced with an equal amount of the oil-controlling and acne-removing composition of example 4; all other components and contents are the same as in application example 1. Application Example 5:

[0094] An application embodiment of the cosmetic described in this invention; the cosmetic is specifically a serum.

[0095] The only difference between this application example and application example 1 is that the oil-controlling and acne-removing composition of example 1 is replaced with an equal amount of the oil-controlling and acne-removing composition of example 5; all other components and contents are the same as in application example 1. Application Example 6:

[0096] An application embodiment of the cosmetic described in this invention; the cosmetic is specifically a serum.

[0097] The only difference between this application example and application example 1 is that 4.2% of the oil-controlling and acne-removing composition of example 1 is replaced with 11.5% of the oil-controlling and acne-removing composition of example 6, while the remaining components and contents are the same as in application example 1.

[0098] Application Example 7:

[0099] An application embodiment of the cosmetic described in this invention; the cosmetic is specifically a serum.

[0100] The only difference between this application example and application example 1 is that 4.2% of the oil-controlling and acne-removing composition of example 1 is replaced with 7.5% of the oil-controlling and acne-removing composition of example 7, while the remaining components and contents are the same as in application example 1.

[0101] Application Comparative Example 1:

[0102] A comparative example of the application of the cosmetic described in this invention; the cosmetic is specifically a serum.

[0103] The only difference between this comparative example and application example 1 is that 4.2% of the oil-controlling and acne-removing composition of example 1 is replaced with 21% of the oil-controlling and acne-removing composition of comparative example 1, while the remaining components and contents are the same as in application example 1.

[0104] Application Comparative Example 2:

[0105] A comparative example of the application of the cosmetic described in this invention; the cosmetic is specifically a serum.

[0106] The only difference between this comparative example and Application Example 1 is that the oil-controlling and acne-removing composition of Example 1 is replaced with an equal amount of the oil-controlling and acne-removing composition of Comparative Example 2; all other components and contents are the same as in Application Example 1. Comparative Example 3:

[0107] A comparative example of the application of the cosmetic described in this invention; the cosmetic is specifically a serum.

[0108] The only difference between this comparative example and Application Example 1 is that the oil-controlling and acne-removing composition of Example 1 is replaced with an equal amount of the oil-controlling and acne-removing composition of Comparative Example 3; all other components and contents are the same as in Application Example 1. Application Comparative Example 4:

[0109] A comparative example of the application of the cosmetic described in this invention; the cosmetic is specifically a serum.

[0110] The only difference between this comparative example and Application Example 1 is that the oil-controlling and acne-removing composition of Example 1 is replaced with an equal amount of the oil-controlling and acne-removing composition of Comparative Example 4; all other components and contents are the same as in Application Example 1. Test Example 1: Performance Evaluation of White Willow (SALIXALBA) Bark Extract

[0111] Test sample: Unmodified / enzyme-modified white willow bark extract from the oil-controlling and acne-removing compositions described in Examples 1-4.

[0112] 1. After obtaining the white willow bark extract, it was validated using HPLC.

[0113] (1.1) Instruments and reagents

[0114] An Agilent Infinite 1260Ⅱ ultrasonic cleaner with a diode array detector, an Agilent ZORBAX SB-C18 (150mm × 4.6mm, 5μm), and a KQ-250 ultrasonic cleaner (Jiangsu Kunshan Ultrasonic Instrument Factory) were used. Salicin reference standard (99% purity, provided by Sigma-Aldrich, USA), white willow bark extract (produced by Xi'an Haotian Bioengineering Technology Co., Ltd.), acetonitrile (analytical grade), and water (double-distilled water) were also used.

[0115] (1.2) Methods and Results

[0116] (1.2.1) Chromatographic conditions:

[0117] Column: Inertsil ODS-3 (150mm × 4.6mm, 5μm);

[0118] Mobile phase: acetonitrile-water (5:95); flow rate: 1.0 mL / min; detection wavelength: 269 nm; column temperature: 25 °C.

[0119] (1.2.2) Preparation of reference solution: Accurately weigh about 5 mg of salicin reference standard into a 25 mL volumetric flask, add water to dissolve and add to the mark to obtain the reference solution.

[0120] (1.2.3) Preparation of test solution: Accurately weigh about 60 mg of white willow bark extract sample into 50 mL, add about 40 mL of water, sonicate for 30 min, let stand at room temperature, and add water to the mark. Filter through a 0.45 μm microporous membrane to obtain the test solution.

[0121] (1.2.4) Regression Curve and Linear Range: Weigh salicin reference standard and dissolve it in water to prepare a 0.20 mg / mL stock solution. Accurately pipette this stock solution to prepare concentrations of 0.04, 0.10, 0.20, 0.30, and 0.40 mg / mL. Perform chromatographic analysis using 10 L of each concentration under the above-described chromatographic conditions. Plot a standard curve with peak area as the ordinate and salicin concentration as the abscissa. Salicin exhibits good linearity in the range of 0.04–0.4 mg / mL, with a linear regression equation of Y = 1.6798X - 1.2923 and r = 0.9996.

[0122] (1.2.5) Precision test: The salicin reference solution was precisely pipetted and injected 5 times consecutively, with 10 μL injected each time. The peak area was measured and the RSD was calculated to be 0.24% (n=5).

[0123] (1.2.6) Reproducibility test: Accurately weigh an appropriate amount of white willow bark extract sample from the same batch, prepare a test solution, inject the sample, and determine 5 samples. The results showed that the average content of salicin was 19.88%, and the RSD was 0.25%.

[0124] (1.2.7) Stability test: The test solution of white willow bark extract was accurately pipetted and injected every 1 hour, with an injection volume of 10 μL. The peak area was measured, and the RSD of salicin peak area was 0.36% (n=9). The results indicate that the test solution is basically stable within 8 hours.

[0125] (1.2.8) Recovery test: The sample of white willow bark extract with a salicin content of 19.50% was accurately weighed, and 0.15, 0.25, 0.35, 0.45 and 0.55 mg of salicin reference standard were accurately added respectively to prepare the test solution, injected and determined. The results showed that the average recovery rate of salicin was 99.35% and the RSD was 0.39% (n=5).

[0126] (1.2.9) Quantify the content of salicylic acid glycosides in the extract of white willow bark for subsequent testing. The test method is based on Zhao Yongqiang et al.'s "Determination of salicylic acid glycoside content in white willow bark by UPLC".

[0127] Sample determination: Accurately weigh the white willow bark extract sample, prepare the test solution, inject the sample, and determine the results. The results showed that the salicylic acid content in the unmodified white willow bark extract was 20.54%, with an RSD of 0.24%; the salicylic acid content in the β-glucosidase-modified white willow bark extract was 25.84%, with an RSD of 0.24%; the salicylic acid content in the β-glucanase-modified white willow bark extract was 22.17%, with an RSD of 0.24%; and the salicylic acid content in the xylanase-modified white willow bark extract was 20.98%, with an RSD of 0.24%.

[0128] 2. Effects of white willow bark extract on the survival rate of SZ95 cells

[0129] Human sebaceous gland cells SZ95 were provided by Shanghai Skin Disease Hospital, and the culture medium used was a medium containing 10% CCK-8 reagent (Dojin Chemical Research Institute, Japan). Cells frozen in liquid nitrogen were thawed, digested, and then passaged normally for experimental use.

[0130] SZ95 cells were seeded at a density of 20,000 cells per well in a 96-well plate and incubated for 24 h in a cell culture incubator. Then, test solutions containing 25, 50, 100, and 200 μg / mL of the drug (salicylic acid) were added, and incubation continued for another 24 h. The culture medium was removed and the cells were washed with phosphate-buffered saline (PBS). Then, culture medium containing 10% CCK-8 reagent was added and incubated for 30 min. Finally, the optical density (OD) of each well at 450 nm was measured using a microplate reader (Molecular Devices, USA), and the cell viability was calculated according to formula (1).

[0131] Cell viability (%) = (OD of the drug-treated group) 450 -PBS well OD 450 ) / (Blank group OD 450 -PBS well OD 450 )×100%——Formula (1)

[0132] The results are as follows Figure 1 As shown. The white willow bark extracts after unmodification (Example 1), β-glucosidase modification (Example 2), β-glucanase modification (Example 3), and xylanase modification (Example 4) all promoted the growth of SZ95 cells (P < 0.05, all showing significant differences). Furthermore, the promoting effect tended to increase with increasing test solution concentration, indicating that the white willow bark extracts had no cytotoxicity to SZ95 cells.

[0133] 3. Effects of white willow bark extract on neutral lipid synthesis in SZ95 cells

[0134] SZ95 cells were seeded at a density of 10,000 per well in BeyoGold 96-well black transparent-bottom cell culture plates (Beyotime Biotechnology Co., Ltd.). The next day, test solutions containing 200 μg / mL of the drug (salicylic acid) were added to each well, and the cells were incubated at 37°C in a 5% CO2 cell culture incubator for 24 h. After aspirating the supernatant and washing with PBS, 10 μg / mL of Nile Red solution was added to each well for staining, and the cells were incubated at 37°C in the dark for 10 min. After washing with PBS, the fluorescence intensity of Nile Red was detected using a microplate reader at an excitation wavelength of 485 nm and an emission wavelength of 565 nm. The relative lipid content in SZ95 cells was expressed as a percentage ratio of fluorescence intensity between the drug-treated group and the control group.

[0135] The results are shown in Table 1.

[0136] Table 1

[0137] White willow bark extract Relative content of neutral lipids (%) β-glucosidase modification 60.63 β-glucanase modification 73.55 Xylanase modification 82.82 Unmodified 84.66

[0138] The results showed that the enzyme-modified white willow bark extract, especially the β-glucosidase-modified extract, significantly improved the inhibition of neutral lipid synthesis in SZ95 cells (P < 0.001, indicating a significant difference), which will provide a good foundation for oil control in the long run.

[0139] 4. Determination of the antibacterial effect of white willow bark extract against Staphylococcus aureus and Propionibacterium acnes

[0140] The antibacterial rate of white willow bark extract against Staphylococcus aureus and Propionibacterium acnes was tested using a micro-broth dilution method.

[0141] The specific experimental procedures are as follows: ① Under aseptic conditions, add 100 μL of nutrient broth (purchased from Qingdao Haibo Biotechnology Co., Ltd., for Staphylococcus aureus culture) or brain heart infusion broth (purchased from Qingdao Haibo Biotechnology Co., Ltd., for Propionibacterium acnes culture) to each well of a 96-well plate; ② Add 100 μL of the 1% (v / v) test solution to well 1, mix it with the culture medium in the well by pipetting, then aspirate 100 μL of the suspension and add it to well 2, and so on until well 11. Then aspirate 100 μL of the suspension from well 11 and discard it. Well 12 is left untreated as a positive control; ③ Add 100 μL of diluted bacterial solution to each well, ensuring that the final concentration of the bacterial solution in each well is 1 × 10⁻⁶. 6 CFU / mL, and a negative control was set up (containing only the test solution and culture medium at different dilutions to remove background OD of the test sample). 630 ); ④ OD is detected after 24 hours of incubation. 630 And calculate the bacterial survival rate and antibacterial rate according to formulas (2) and (3).

[0142] Bacterial survival rate (%) = (OD of the treatment group) 630 - Negative control well OD 630 ) / (Positive control well OD 630

[0143] - Negative control well OD 630 )×100%——Formula (2)

[0144] Inhibition rate (%) = 1 - Bacterial survival rate (%) - Equation (3)

[0145] The extract of white willow bark was diluted 4-64 times, and its inhibitory effect on Staphylococcus aureus was observed. Figure 2 It can be seen that the white willow bark extracts of each embodiment have good antibacterial effects. Among them, the white willow bark extract modified by β-glucosidase has the best antibacterial rate against Staphylococcus aureus.

[0146] The extract of white willow bark was diluted 4-64 times, and its inhibitory effect on Propionibacterium acnes was observed. Figure 3 It can be seen that the white willow bark extracts of each embodiment have good antibacterial effects. Among them, the white willow bark extract obtained by β-glucosidase modification (Example 2) has the best antibacterial rate against Propionibacterium acnes.

[0147] Test Example 2: Oil Control and Acne Removal Effect Test

[0148] Test samples: cosmetics (serums) prepared using Examples 1-7 and Comparative Examples 1-4.

[0149] 1. Select volunteers to conduct product testing and evaluation.

[0150] All enrolled participants met the following inclusion and exclusion criteria:

[0151] (1.1) Subject inclusion criteria:

[0152] (1.1.1) 18 to 65 years old, healthy female or male;

[0153] (1.1.2) Facial skin has problems such as excessive oil production, redness, and acne;

[0154] (1.1.3) People with sensitive skin (through a sensitive skin questionnaire and a lactic acid stinging score ≥3 points);

[0155] (1.1.4) The test subjects have not participated in other clinical studies in the past two months, and the test sites have not undergone any skin treatments, cosmetic procedures, or other tests that may affect the test results.

[0156] (1.1.5) Those who can understand the trial process, voluntarily participate in the trial, and sign a written informed consent form.

[0157] (1.2) Exclusion criteria for subjects:

[0158] (1.2.1) Pregnant or breastfeeding women or those who plan to conceive in the near future;

[0159] (1.2.2) Those with a history of skin diseases such as psoriasis, eczema, atopic dermatitis, or severe acne; or those with other chronic systemic diseases;

[0160] (1.2.3) Individuals who have taken orally or used topical anti-inflammatory drugs such as corticosteroids within the past month;

[0161] (1.2.4) Individuals with severe allergies;

[0162] (1.2.5) Individuals who have used retinoid preparations or undergone cosmetic treatments such as chemical peels, lasers, or pulsed light on the test site within the past 3 months;

[0163] (1.2.6) Those who are unavoidably exposed to sunlight for extended periods;

[0164] (1.2.7) Individuals who have participated in other clinical trials within the past two months;

[0165] (1.2.8) Other clinical assessments deemed unsuitable for participation in the trial.

[0166] (1.3) The subject information selected for this test case is shown in Table 2.

[0167] Table 2 Subject Information

[0168] Human body testing Total number of people male female Participants (total number) 32 0 32 Subjects (Number of Valid Participants) 32 0 32

[0169] 2. Testing instruments and materials

[0170] VISIA7 (PIMIA analysis system), Mexameter MX18, Tewameter™ HEX, Corneometer CM825, Primos CR.

[0171] 3. Test location and environment

[0172] Testing Location: Efficacy Evaluation Laboratory, Guangzhou Zhongke Testing Technology Co., Ltd.

[0173] Test environment: Temperature 21±1℃; Humidity 50±10%.

[0174] 4. Testing Methods

[0175] (4.1) The test items are shown in Table 3:

[0176] Table 3 Test Items

[0177]

[0178]

[0179] (4.2) Basic Principles

[0180] Before human testing of cosmetic products, necessary toxicological tests must be completed and written certification issued. Samples that fail toxicological tests will not be tested on humans. According to the Declaration of Helsinki, the selection of subjects follows medical and ethical standards for human testing. All subjects must participate voluntarily and sign an informed consent form before the test. Before signing the informed consent form, the testing personnel must inform the subject of the purpose of the test, the potential benefits, potential risks and problems, and the related rights and obligations.

[0181] (4.3) Testing process

[0182] (4.3.1) Product usage methods and frequency

[0183] After cleansing your face in the morning and evening, take an appropriate amount of this product and apply it to the pimples.

[0184] (4.3.2) Test Procedure

[0185] Subjects were recruited and enrolled according to the requirements, and a written informed consent form was signed. Before enrollment, subjects were asked a series of questions about their medical history and health status based on the inclusion and exclusion criteria. Then, a lactic acid stinging test was performed (see 4.3.2.1 for the specific testing procedure) to select subjects who met the criteria to participate in this testing project.

[0186] Participants cleansed their faces with facial cleanser, rinsed thoroughly with water, and then dried their faces with lint-free absorbent facial tissues. After cleansing, they entered a temperature and humidity controlled chamber and waited for 30 minutes. During this stabilization period, participants were not allowed to eat or drink, their foreheads were exposed, and they were kept relaxed and avoided touching the test sites.

[0187] After the rest period, the baseline skin values ​​were tested, and the skin testing procedure (see 4.3.2.2 for details) and the VISIA imaging testing procedure (see 4.3.2.3 for details) were completed. The test site was the acne-prone area on the face. The subjects were informed to use the test sample according to the sample usage instructions.

[0188] (4.3.2.1) Lactic acid stinging test

[0189] The lactic acid stinging test was used to assess whether subjects had sensitive skin. 50 μL of 10% lactic acid solution was placed on a 1*1 cm filter paper, which was then applied to one side of the subject's nasolabial fold, with a saline control on the other side. The tester asked subjects about their stinging sensation at 0.5 min, 2.5 min, and 5 min, and scored them using the "4-point scale" in the table below. Subjects with a cumulative stinging sensation ≥3 points on the lactic acid side compared to the saline side were considered to have sensitive skin. The lactic acid stinging score levels are shown in Table 4.

[0190] Table 4

[0191] Grade Score Scoring criteria 0 points No stinging sensation 1 point Mild tingling sensation: mild and perceptible 2 points Moderate stinging sensation: noticeably tolerable 3 points Severe stinging sensation: Extremely stinging, unbearable

[0192] (4.3.2.2) Skin testing procedure

[0193] Skin moisture content, transepidermal water loss (TEWL), and skin hemoglobin were measured using a skin analyzer at 0h, 48h, and 7 days, with a test area of ​​3cm×3cm. The results were recorded and statistically analyzed.

[0194] (4.3.2.3) VISIA Inspection Process

[0195] The area of ​​acne erythema, skin pore density, and skin comedones were measured using a VISIA instrument at 0h, 48h, and 7 days, and the data were recorded and statistically analyzed.

[0196] (4.4) Data Analysis

[0197] Statistical analysis software was used to perform statistical analysis on the data. Quantitative data were expressed as mean ± standard deviation and tested for normality. If the data conformed to a normal distribution, paired t-tests were used for comparisons of the data before and after the test; otherwise, rank-sum tests of two related samples were used. Ordinal data were compared using rank-sum tests of two related samples. Comparisons between the experimental sample and the control group were performed using independent samples t-tests or rank-sum tests. All statistical analyses were two-tailed tests, with a significance level of α = 0.05.

[0198] (4.5) Criteria for Judging Test Conclusions

[0199] (4.5.1) Claiming acne-removing efficacy: Using a self-comparison before and after, observe the area of ​​skin erythema, Pillsbury score, skin pore density, acne volume, and transepidermal water loss rate. If at any visit time point after using the sample, the test values ​​of 3 or more parameters are better than the baseline values ​​before use, and the results are significantly different (P<0.05), then the test sample is considered to have acne-removing efficacy.

[0200] ① Skin erythema area: At each visit point, the skin erythema area was collected using a VISIA instrument. The smaller the measured value, the smaller the skin erythema area.

[0201] ②Pillsbury rating scale: At each visit point, professionals scored the acne lesions, inflammatory papules, and pustules on the subject's face.

[0202] Grade 1 (mild): A small number of pimples and papules, without inflammation;

[0203] Grade 2 (Moderate): Numerous comedones and papules, with a few inflammatory papules or pustules;

[0204] Grade 3 (Moderate to Severe): Numerous inflammatory papules and pustules, possibly with a few nodules;

[0205] Grade 4 (Severe): Numerous inflammatory papules, pustules, and nodules, possibly with cysts.

[0206] ③ Skin pore density measurement: At each visit point, the skin pore density was collected and analyzed using a VISIA instrument. The smaller the measured value, the less skin pore density there is.

[0207] ④ Acne volume: At each visit point, the acne volume was collected using a Primos CR instrument. The smaller the measured value, the smaller the acne volume.

[0208] ⑤ Measurement of transepidermal water loss rate: At each visit point, the transepidermal water loss rate of the skin was collected and analyzed using the Tewameter™ HEX instrument. The smaller the measured value, the better the skin barrier.

[0209] (4.5.2) Claims of soothing effects: Using a self-comparison before and after, observe the area of ​​skin erythema, skin TEWL, and skin hemoglobin. The smaller the measured values ​​of these three parameters, the smaller the area of ​​skin erythema, the better the skin barrier, and the weaker the redness. If, at any visit time point after using the sample, the test values ​​of two of these parameters are better than the baseline values ​​before use, and the results are statistically significant (P<0.05), then the test sample is considered to have a soothing effect.

[0210] (4.5.3) Claims of repair efficacy: Using a self-comparison before and after, observe that the lower the measured values ​​of skin TEWL and hemoglobin, the less transepidermal water loss and the weaker the redness of the skin, respectively. If the test values ​​of both are better than the baseline values ​​before use at any time point after using the sample, and the results are significantly different (P<0.05), then the test sample is considered to have the effect of improving pores.

[0211] (4.5.4) Claiming a mild and non-irritating effect: Using a self-comparison before and after, observe the area of ​​skin erythema and skin hemoglobin. The smaller the measured values ​​of both, the smaller the area of ​​skin erythema and the weaker the degree of redness. If the measured values ​​of both are better than the baseline values ​​before use at any time point after using the sample, and the results are significantly different (P<0.05), then the test sample is considered to have a mild and non-irritating effect.

[0212] The actual performance of the product is shown in Table 5-6.

[0213] Table 5 shows the 7-day efficacy test data for human subjects:

[0214]

[0215]

[0216]

[0217]

[0218] Table 6. Statistics on adverse skin reactions

[0219]

[0220] 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 oil-controlling and acne-removing composition, characterized in that, It consists of the following components in parts by weight: 2 parts capryloyl salicylic acid, 1 part magnolol, and 1.2 parts white willow bark extract; The preparation method of the white willow bark extract includes the following steps: (1) Water was added to the bark of white willow to obtain an aqueous extract of white willow bark. The extract was concentrated, and heterobranching acid synthase was added to assist the extraction. Ethanol was added while stirring. The extract was then allowed to stand in a sealed container, and the ethanol was recovered by filtration. The extract was dried to obtain a powder, and water was added to prepare an aqueous solution. The content of salicylic acid in the powder of the white willow bark aqueous extract was >18%. (2) Add β-glucosidase to the aqueous solution obtained in step (1) for modification treatment, concentrate, and freeze dry to obtain the white willow bark extract.

2. The oil-controlling and acne-removing composition as described in claim 1, characterized in that, In step (2), the concentration of the enzyme is 500-800 U / mL.

3. The oil-controlling and acne-removing composition as described in claim 1, characterized in that, In step (2), the conditions for enzyme modification treatment are: adjusting the pH to 5.0-6.0 and treating at 50-60℃ for 1-3 h.

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

5. A cosmetic product, characterized in that, The cosmetic product includes the oil-controlling and acne-removing composition according to any one of claims 1-3.

Images