Composition with long-acting acne removing effect and application thereof

Abstract

The invention discloses a composition with a long-acting acne removing effect and application of the composition. The acne removing composition is prepared from hylocereus undatus fruit extract and a fermentation composition, and the fermentation composition is prepared from glycolipid and a lactobacillus fermentation product; the mass ratio of the hylocereus undatus fruit extract to the fermentation composition is (1-4): (5-2); the glycolipid is sophorolipid. The composition has a synergistic effect of inhibiting propionibacterium acnes, can realize long-acting acne removal and reduce the recurrence rate, and has a good acne removal effect when being applied to shampoo, hair care products or scalp care products.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetics, and particularly relates to a composition with long-lasting acne-removing effect and application thereof. Background Art

[0002] Acne, also known as pimples, is a common skin problem that usually occurs during adolescence and also occurs in adults. Propionibacterium acnes (scientific name: Propionibacterium acnes) is a species of the genus Propionibacterium and is the pathogen that causes acne. With the accelerated pace of life and the increase of social pressure, bad living habits, such as high-fat and high-sugar diets, staying up late, and mental stress, may lead to the occurrence of acne. Acne not only brings discomfort symptoms such as pain, but also affects appearance and self-confidence, brings negative emotions to patients, and seriously affects the quality of life.

[0003] The acne removal effect of current acne removal products is still limited, and they are prone to relapse, making it difficult to achieve long-term acne removal. Therefore, the development of long-term acne removal products is a technical problem to be solved and has broad market prospects. Summary of the invention

[0004] The purpose of this section is to summarize some aspects of embodiments of the present invention and to briefly introduce some preferred embodiments.

[0005] As one aspect of the present invention, the present invention provides a composition with long-lasting acne-removing effect, wherein: the acne-removing composition is composed of a Cynomorium orchioides fruit extract and a fermentation composition, and the fermentation composition is composed of glycolipids and lactobacillus fermentation products; the mass ratio of the Cynomorium orchioides fruit extract to the fermentation composition is (1-4): (5-2); and the glycolipid is a sophorolipid.

[0006] As a preferred embodiment of the composition with long-lasting acne-removing effect of the present invention, the mass ratio of the glycolipid to the lactobacillus fermentation product is (65-70): (30-35).

[0007] As a preferred embodiment of the composition with long-lasting acne-removing effect described in the present invention: the sophorolipid comprises a lactone-type sophorolipid.

[0008] As a preferred embodiment of the composition with long-term acne-removing effect of the present invention: the preparation method of the glycolipid comprises: fermenting Candida in a fermentation medium, standing and stratifying after the fermentation, and collecting the lactone-type sophorolipid in the bottom layer.

[0009] As a preferred embodiment of the composition with long-lasting acne-removing effect of the present invention, it also includes extracting the collected bottom lactone-type sophorolipid.

[0010] As a preferred embodiment of the composition with long-lasting acne-removing effect described in the present invention: the preparation method of the Hylocereus orientalis fruit extract comprises squeezing Hylocereus orientalis fruit to obtain fruit pulp, and filtering to obtain the Hylocereus orientalis fruit extract.

[0011] Preparation method of composition with long-lasting acne-removing effect:

[0012] (1) Preparation of glycolipids: The glycolipids used in the present invention are sophorolipids (or Candida albicans / glucose / rapeseed oleic acid methyl ester fermentation products). Sophorolipids are microbial secondary metabolites produced by Candida albicans using sugar and vegetable oil as carbon sources through a fermentation process under certain conditions. The sophorolipids of the present invention are mainly lactone-type sophorolipids.

[0013] (2) Preparation of lactobacillus fermentation products:

[0014] The lactobacillus is inoculated into the culture medium for activation culture, the activated lactobacillus is inoculated into the culture medium, fermented at 25-40°C for 48-72 hours, and then lysozyme is added for enzymolysis, filtered, sterilized, and a lactobacillus fermentation product is obtained.

[0015] (3) Preparation of the extract of Hylocereus orientalis fruit:

[0016] Fresh Hylocereus orientalis fruits are picked, washed, chopped, and then ground into pulp by a pulping machine, and then put into a stirring device for stirring at a speed of 4000-6000 rpm, and pressed, the pressure is set to 65-85 MPa, and the pressure holding time is 30-60 minutes. After the pressing is completed, the Hylocereus orientalis fruit liquid obtained by pressing is filtered, and the pore size of the filter membrane is 250-450 nm to obtain the Hylocereus orientalis fruit extract.

[0017] (4) The glycolipid obtained in step (1) and the lactobacillus fermentation product obtained in step (2) are mixed in a mass ratio of (65-70):(30-35), and stirred evenly to obtain a fermentation composition.

[0018] (5) mixing the fermented composition obtained in step (4) and the Cyperus rotundus fruit extract obtained in step (3) in a certain mass ratio, stirring evenly, to obtain an anti-acne composition.

[0019] Beneficial effects of the invention: The invention discloses a long-acting acne-removing composition, which has the effect of synergistically inhibiting Propionibacterium acnes, and can achieve long-acting acne removal and reduce the recurrence rate. It has a good acne-removing effect when used in shampoo, hair care products or scalp care products. DETAILED DESCRIPTION

[0020] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with specific embodiments.

[0021] Embodiment 1:

[0022] The anti-acne composition of the present invention is composed of raw material 1, raw material 2 and raw material 3, and the specific components are as follows:

[0023] Raw material 1: glycolipid, or fermentation product of Candida albicans / glucose / rapeseed oleic acid methyl ester;

[0024] Ingredient 2: INCI name: Lactobacillus fermentation product;

[0025] Ingredient 3: INCI name: HYLOCEREUS UNDATUS fruit extract.

[0026] (1) Preparation of sophorolipids:

[0027] The glycolipid used in the present invention is a sophorolipid, which is a microbial secondary metabolite produced by Candida using sugar and vegetable oil as carbon sources through a fermentation process under certain conditions.

[0028] Bacteria cultivation and fermentation method: YPD solid culture medium (g / L): yeast powder 10, peptone 20, glucose 20, agar 20, high pressure sterilization at 115℃ for 35min.

[0029] Fermentation medium (g / L): glucose 100, yeast powder 5, KH2PO4 1, MgSO4·7H2O 0.5, NaCl 0.1, CaCl2·2H2O 0.1, peptone 0.7, initial rapeseed oil 50, high pressure sterilization at 121℃ for 20min.

[0030] Plate / slant culture: Activate the strain, pick the Candida strain (ATCC 22214) and streak it on a YPD solid medium plate / test tube slant, culture at 30°C for 48 hours to obtain a single colony.

[0031] Seed liquid culture: Pick a single colony from the plate and inoculate it into 50 mL YPD liquid medium, and culture it at 30°C and 220 rpm in a shaking incubator for 48 h.

[0032] Batch feeding fermentation in a 5L fermenter: The liquid volume of the 5L fermenter is 2L, the rotation speed is controlled at 350-450rpm, the temperature is maintained at 30°C, and the ventilation volume is maintained at 3-6L / min. Use NaOH to maintain the pH between 3.5-3.8 after 24 hours before fermentation. Take samples at regular intervals after inoculation to measure the bacterial concentration, sophorolipid concentration and residual sugar concentration. The pH and dissolved oxygen can be directly read through the electrode indication on the fermenter. When the glucose content is lower than 20g / L, feed with 80% (w / v) glucose aqueous solution to 20g / L or more; when the rapeseed oil content is lower than 30g / L, feed to 30g / L or more.

[0033] Layered collection of sophorolipids: After the fermentation is completed, the fermentation liquid is first loaded into a separatory funnel, and allowed to stand overnight to naturally settle and separate into layers to obtain the upper layer of residual oil (which can be recycled), the middle layer of bacterial fermentation liquid mainly composed of acid-type sophorolipids, and the bottom layer of sophorolipids mainly composed of lactone-type sophorolipids.

[0034] Ethanol extraction to remove water-soluble macromolecular impurities: Separate and collect a certain volume of brown-yellow oily viscous lactone-type sophorolipids, add three times the volume of anhydrous ethanol and mix well, stir for 30 minutes, and flocculent protein and residual sugar can be seen precipitated. Centrifuge at 8000rpm for ten minutes to remove flocculent impurities, and obtain a clear brown-yellow glycolipid ethanol solution on the upper layer.

[0035] Removal of ethanol by rotary evaporation: At 65°C, the glycolipid ethanol solution was rotary evaporated to remove the solvent ethanol and obtain a viscous, clear, brown-yellow lactone-type sophorolipid.

[0036] (2) Preparation of lactobacillus fermentation products:

[0037] Lactobacillus plantarum (Lactobacillus p / antarum) is inoculated into a solid culture medium for activation culture. The activated lactobacillus is inoculated into the culture medium and fermented at 37°C for 48 hours. Then, 0.1% lysozyme is added and enzymolysis is performed at 35°C for 1 hour. After the enzymolysis is completed, the product is filtered and pasteurized to obtain a lactobacillus fermentation product.

[0038] (3) Preparation of the extract of Hylocereus orientalis fruit

[0039] Fresh Hylocereus orientalis fruits are picked, washed, chopped, and then ground into pulp by a pulping machine, and then put into a stirring device for stirring at a stirring speed of 4000-6000rpm. After the feed valve of the squeezer is opened, the feed pump presses and feeds the Hylocereus orientalis pulp, and the feed pressure is 0.3MPa. The feed valve of the equipment is closed, and squeezing is performed, and the pressure is set to 65-85MPa, and the pressure holding time is 30-60min. After squeezing is completed, the Hylocereus orientalis fruit liquid obtained by squeezing is filtered multiple times, and the filter membrane pore size is 250-450nm. Preferably, the squeezing pressure is set to 80MPa, the pressure holding time is 40min, and the filter membrane pore size is 400nm.

[0040] (4) The glycolipid obtained in step (1) and the lactobacillus fermentation product obtained in step (2) are mixed in a mass ratio of (65-70):(30-35) (preferably in a mass ratio of 68:32), and stirred evenly to obtain a fermentation composition.

[0041] (5) The fermentation composition obtained in step (4) (composite raw material of glycolipid and lactobacillus fermentation product) and the extract of the fruit of the radix serrata obtained in step (3) are mixed in a certain mass ratio and stirred evenly to obtain an anti-acne composition.

[0042] Inhibition test on Propionibacterium acnes and verification of synergistic effect:

[0043] Minimum inhibitory concentration test:

[0044] In this test, the agar dilution method was used to mix and dissolve different concentrations of antibacterial agents in agar medium, and then the bacteria were inoculated. The minimum concentration of antibacterial (inhibitory) substances that inhibited the growth of the test bacteria was determined by the growth of the bacteria. The minimum concentration that inhibited the growth of the test bacteria is the minimum inhibitory concentration (MIC).

[0045] (1) Preparation of antibacterial (inhibitory) solution: 32 mg of the sample to be tested was taken by aseptic operation, placed in a sterile phosphate buffer, and fully shaken to dissolve to prepare a 32 mg / mL uniformly dispersed solution. The sample to be tested is the extract of the fruit of the genus Cyperus and the fermentation composition (a combination of glycolipid and lactobacillus fermentation product).

[0046] (2) Preparation of culture medium containing antimicrobial agent: dilute the antimicrobial (inhibitory) solution prepared at 32 mg / mL with PBS in multiples to prepare test solutions of different concentrations, and place in a water bath at a constant temperature of 45°C to 50°C for later use.

[0047] (3) Preparation of double concentration culture medium: Weigh 76g MH agar medium and dissolve it in 1000mL water. Heat until boiling and dissolved. Then sterilize it with 121℃ pressure steam for 15min and place it in a 45℃~50℃ water bath for later use. This culture medium will be used to dilute the antibacterial (inhibitory) solution or suspension.

[0048] (4) Preparation of culture medium containing antibacterial (inhibitory) solution: Take 10 mL of serially diluted antibacterial solution and add it to the plate. Add 10 mL of double MH agar medium in a 45℃~50℃ water bath to the plate, shake the plate while adding to mix the antibacterial (inhibitory) solution and culture medium thoroughly, and wait for solidification before use.

[0049] (5) Take 2 μl (containing about 10 7 cfu / ml) of Propionibacterium acnes suspension was inoculated on a plate containing an antibacterial (inhibitory) culture medium. The diameter of the bacterial liquid circle formed after inoculation was about 5mm to 8mm (the amount of bacteria in each spot was about 10 4 cfu).

[0050] (6) Inoculate MH agar plates without antibacterial (inhibitory) ingredients in the same way as a positive control.

[0051] (7) Place the inoculated plate in a 35°C incubator for 48 hours and observe the results.

[0052] The minimum inhibitory concentration experimental records and results are shown in Table 2:

[0053] Table 1

[0054]

[0055] Note: “+” indicates that the colony growth is completely inhibited; “-” indicates the colony growth.

[0056] Verification of the synergistic effect of the composition:

[0057] In this experiment, the checkerboard method was used for design, and the agar dilution method was used to determine the minimum inhibitory concentration of the Hylocereus undatus fruit extract and the fermentation composition (a compound of glycolipid and Lactobacillus fermentation product) at different concentrations against Propionibacterium acnes. Taking the fractional inhibitory concentration (FIC) index as an indicator, FIC index = MIC of the combination of drug A / MIC of drug A used alone + MIC of the combination of drug B / MIC of drug B used alone. FIC ≤ 0.5 indicates a synergistic effect, 0.5 < FIC ≤ 1 indicates an additive effect, 1 < FIC ≤ 2 indicates an irrelevant effect, and FIC > 2 indicates an antagonistic effect.

[0058] To explore the combined antibacterial effect of the Hylocereus undatus fruit extract and the fermentation composition (a compound of glycolipid and Lactobacillus fermentation product). Test method: The 2-fold of the MIC of the Hylocereus undatus fruit extract and the fermentation composition (a compound of glycolipid and Lactobacillus fermentation product) against Propionibacterium acnes was taken as the highest dilution concentration respectively, and 9 series of dilution concentrations were prepared respectively according to the serial dilution method. The prepared Hylocereus undatus fruit extract and fermentation composition (a compound of glycolipid and Lactobacillus fermentation product) at different concentrations were combined in pairs according to the checkerboard method and added into test tubes, 0.5 ml of each drug. Keep it in a water bath at 45°C - 50°C for constant temperature for standby. Subsequently, the following steps refer to steps (3)-(7) of the minimum inhibitory concentration test.

[0059] Table 2 Results of the checkerboard microdilution method

[0060]

[0061] Note: “+” indicates that the colony growth is completely inhibited; “-” indicates the colony growth.

[0062] Table 3

[0063]

[0064] As can be seen from Table 3 and Table 4, the minimum inhibitory concentrations of the combination of the Hylocereus undatus fruit extract and the fermentation composition (a compound of glycolipid and Lactobacillus fermentation product) against Propionibacterium acnes are 2 mg / mL and 2 mg / mL respectively, and the FIC value is 0.375 < 0.5; the Hylocereus undatus fruit extract and the fermentation composition (a compound of glycolipid and Lactobacillus fermentation product) have a synergistic inhibitory effect on Propionibacterium acnes.

[0065] Prepare the anti-acne compositions according to Table 4 for Examples 1 - 4 and Comparative Examples 1 - 2.

[0066] Table 4

[0067]

[0068] A scalp essence containing the anti-acne composition of Examples 1-4 or Comparative Examples 1-2 was prepared, and the composition was as follows according to mass percentage:

[0069]

[0070] The preparation method of the scalp essence containing the acne-removing composition is as follows:

[0071] (1) Take a beaker and mix 1,3-butanediol and 1,2-hexanediol with an appropriate amount of deionized water and stir evenly;

[0072] (2) Heat PEG-40 hydrogenated castor oil at 45°C in another beaker to dissolve it, add the anti-acne composition and stir evenly; continue to add 95% ethanol and stir evenly;

[0073] (3) Add the solution in (2) to (1), stir evenly, and take a sample to obtain the finished product.

[0074] In order to verify the efficacy of the acne-removing combination, comparative experiments were conducted based on the comparative examples and embodiments:

[0075] Table 5

[0076]

[0077] Acne test:

[0078] Test method: 120 subjects, with acne or pimples on the scalp or hairline, aged 20-45 years old, no statistically significant differences in gender, average age, and EASI score, were randomly divided into 6 groups, 20 people in each group. The above-mentioned examples and comparative examples were used respectively. The frequency of use was twice a day. During the test, the distributed basic shampoo was used, and no other scalp care products were used, and no beauty and hairdressing behavior was performed. After using the product for 4 weeks, the acne removal effect of the product was evaluated.

[0079] Cured: The pimples or blackheads disappear.

[0080] Effective: Acne or blackheads are visibly reduced.

[0081] Ineffective: The pimples or blackheads do not disappear or the symptoms get worse.

[0082] Recurrence: Acne or blackheads still appear within 14 days after treatment.

[0083] Effective rate: (cured + effective) / total number of patients X 100%.

[0084] Relapse rate: relapse / (recovery+effectiveness)X100%.

[0085] Table 6 Acne removal test results

[0086]

[0087]

[0088] From the results in Table 6, it can be seen that the mass ratio of the extract of the fruit of the radix ophiopogonis and the fermentation composition (a combination of glycolipids and lactobacillus fermentation products) is (1-4): (5-2), when used in scalp essence, can help remove acne or blackheads, and reduce the recurrence rate of acne or blackheads, and has a long-lasting acne removal effect.

[0089] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. A composition with long-lasting acne-removing effect, characterized in that: The acne-removing composition is composed of a physalis extract and a fermentation composition, wherein the fermentation composition is composed of glycolipids and lactobacillus fermentation products; the mass ratio of the physalis extract to the fermentation composition is (1-4):(5-2); and the glycolipid is a sophorolipid.

2. The composition with long-lasting acne-removing effect according to claim 1, characterized in that: The mass ratio of the glycolipid to the lactobacillus fermentation product is (65-70):(30-35).

3. The composition with long-lasting acne-removing effect according to claim 1 or 2, characterized in that: The sophorolipids include lactone-type sophorolipids.

4. The composition with long-lasting acne-removing effect according to claim 3, characterized in that: The preparation method of the glycolipid comprises: fermenting Candida in a fermentation medium, standing and stratifying after the fermentation is completed, and collecting the lactone-type sophorolipid in the bottom layer.

5. The composition with long-lasting acne-removing effect according to claim 4, characterized in that: The method also includes extracting the collected bottom layer lactone-type sophorolipid.

6. The composition with long-lasting acne-removing effect according to claim 1 or 2, characterized in that: The preparation method of the Hylocereus orientalis fruit extract comprises squeezing Hylocereus orientalis fruit to obtain fruit pulp, and filtering to obtain the Hylocereus orientalis fruit extract.

7. The composition with long-lasting acne-removing effect according to claim 6, characterized in that: The pressing pressure is 65-85 MPa; the pore size of the filtering membrane is 250-450 nm.

8. Use of the composition with long-lasting acne-removing effect according to claim 1 in cosmetics with acne-removing effect.

9. The use according to claim 8, characterized in that: The cosmetics include one or more of shampoo, hair care products, and scalp care products.

10. The use according to claim 8, characterized in that: The composition with long-lasting acne-removing effect can inhibit Propionibacterium acnes.

Images