Application of manuka honey in skin antibiosis and anti-inflammation

Abstract

The invention provides application of manuka honey in skin antibiosis and anti-inflammation, and belongs to the technical field of cosmetics, the application comprises the following steps: (1) mixing rose petals, perilla roots, grape seeds and manuka honey, and adding the mixture into water to obtain an enzymolysis substrate; (2) sequentially adding pectinase, neutral protease and beta-glucanase into the enzymolysis substrate, and performing enzymolysis to obtain an enzymolysis product; (3) adding lactobacillus brevis, lactobacillus helveticus and bifidobacterium lactis into the enzymolysis product, and fermenting to obtain a fermentation product; (4) adding an ethanol aqueous solution into the fermentation product, stirring, filtering by using a filter membrane to obtain a supernatant, and concentrating the supernatant to obtain a mixed extract; and (5) preparing skin antibacterial and anti-inflammatory products from the mixed extract. The mixed extract disclosed by the invention is good in batch stability, relatively good in antibacterial and anti-inflammatory effects and low in irritation.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetics, and particularly relates to application of Manuka honey in skin antibacterial and anti-inflammatory effects. Background Art

[0002] Manuka Honey is a type of honey originating from New Zealand and other regions. This honey is made by bees from the nectar of the Manuka tree (Leptospermumscoparium, also known as manuka or tea tree). The Manuka tree is mainly distributed in the native forests of New Zealand, and is also distributed in small quantities in parts of Australia. Manuka honey is known for its high content of non-hydrogen peroxide antibacterial substances, which is a unique antibacterial property that is different from ordinary honey. In addition, Manuka honey also contains antioxidant ingredients, which are believed to help support the immune system and promote wound healing.

[0003] Since the production of Manuka honey requires a specific geographical environment and conditions, coupled with its uniqueness, its cost is relatively high, and the price of cosmetics using Manuka honey as a raw material will also increase accordingly. Therefore, manufacturers hope to maximize its role in cosmetics.

[0004] However, in the actual production process, the application of Manuka honey in cosmetics has the following problems: (1) As a natural product, honey may undergo chemical changes over time, which may affect its stability and effect in cosmetics. (2) Although Manuka honey has many benefits, some people may be sensitive or allergic to honey ingredients. How to reduce allergenicity is an urgent problem to be solved. (3) Since Manuka honey is relatively expensive, it is beneficial to increase the sales profit of enterprises to make it play multiple functions.

[0005] The Chinese patent with patent application number 202110993075.4 discloses a honey fermentation product, a skin topical preparation containing the same, and a preparation method and application thereof. The preparation method of the honey fermentation product comprises the following steps: lactic acid bacteria are inoculated into an enzymatic substrate, fermented and cultured, and sterilized. However, the antibacterial stability of the honey fermentation product produced by this method is not good.

[0006] Therefore, there is an urgent need for the application of Manuka honey in skin antibacterial and anti-inflammatory effects. Summary of the invention

[0007] The invention aims to provide an application of Manuka honey in skin antibacterial and anti-inflammatory effects.

[0008] In order to achieve the above object, the present invention provides the following technical solutions:

[0009] An application of Manuka honey in skin antibacterial and anti-inflammatory treatment, the application comprising the following steps:

[0010] (1) mixing rose petals, perilla roots, and grape seeds, and crushing them to a particle size of less than 100 mesh to obtain a powder, pre-treating the powder to obtain a pre-treated powder, mixing the pre-treated powder with Manuka honey, and adding the mixture to water to obtain an enzymatic substrate;

[0011] (2) adding pectinase to the enzymatic substrate, performing enzymatic hydrolysis at 35° C.-38° C. for 60-80 min; inactivating the enzyme, adding neutral protease, performing enzymatic hydrolysis at 35° C.-38° C. for 130-150 min; inactivating the enzyme, continuing to add β-glucanase, performing enzymatic hydrolysis at 35° C.-38° C. for 100-120 min; inactivating the enzyme, and obtaining an enzymatic hydrolysis product;

[0012] (3) adding Lactobacillus brevis to the enzymatic hydrolysate, fermenting at 40° C.-45° C. for 3-4 h, continuing to add Lactobacillus helveticus, fermenting at 40° C.-45° C. for 6-7 h, and finally adding Bifidobacterium lactis, fermenting at 40° C.-45° C. for 4-5 h, sterilizing, and obtaining a fermentation product;

[0013] (4) adding ethanol aqueous solution to the fermentation product, stirring at 200 rpm / min-300 rpm / min for 1-2 h, filtering with a filter membrane, and concentrating the obtained supernatant to obtain a mixed extract;

[0014] (5) The mixed extract is used to prepare skin antibacterial and anti-inflammatory products.

[0015] Furthermore, in step (1), the weight ratio of rose petals, perilla roots, grape seeds and Manuka honey is (2-4): (8-10): 10: (13-15).

[0016] Furthermore, the weight of water added in step (1) is 10-15 times the total weight of rose petals, perilla roots, grape seeds and Manuka honey.

[0017] Furthermore, the step of pretreating the powder in step (1) is as follows: firstly, the powder is placed in a low temperature of -30°C for 2-3 minutes, then the temperature is rapidly raised to 30°C within 2 seconds, and the process is repeated 2-3 times; then the powder is pressurized to 16-18MPa, maintained for 4-5 minutes, and then the pressure is released to normal pressure, and the process is repeated 2-3 times to obtain the pretreated powder.

[0018] The present invention performs enzymolysis and fermentation on Manuka honey to improve the antibacterial and anti-inflammatory effects of the product, but it is found that the antibacterial storage stability of the prepared product is not ideal, and the antibacterial and anti-inflammatory effects decrease after being placed for a period of time. The present invention performs mixed enzymolysis and fermentation of rose petals, perilla roots, grape seeds and Manuka honey in a specific ratio, which can improve the storage stability of the product. It is also found that when rose petals, perilla roots, grape seeds and Manuka honey in a specific ratio are enzymolyzed and fermented, the storage stability at high temperature can also be improved. The analysis shows that more active components can be released by pre-treated rose petals, perilla roots, and grape seeds, and mixed with Manuka honey to enzymolyze and ferment multiple components, which can produce a synergistic effect, that is, the interaction between different components may enhance the overall antibacterial and anti-inflammatory effects, and help prevent the product from deteriorating during long-term storage.

[0019] Furthermore, in the step (2), the dosage of pectinase is 600-800u / mL of enzymatic substrate; the dosage of neutral protease is 1000-1200u / mL of enzymatic substrate; and the dosage of β-glucanase is 1400-1600u / mL of enzymatic substrate.

[0020] Furthermore, in step (3), the amount of Lactobacillus brevis is 10 6 -10 7 / mL enzymatic hydrolysate, the dosage of Lactobacillus helveticus is 10 8 -10 9 / mL enzymatic hydrolysate, the dosage of Bifidobacterium lactis is 10 7 -10 8 / mL enzymatic hydrolysis product.

[0021] During the test, it was found that the product fermented with a single enzyme and bacteria had a relatively strong irritation. The present invention can improve the irritation of the product by first using three enzymes for gradient enzymolysis and then using three bacteria for gradient fermentation. At the same time, the moisturizing performance of the product under this condition is also improved. Analysis shows that more diverse metabolites can be produced under this condition, and these products can play a neutralizing role, reduce the concentration of certain irritating substances, thereby reducing the irritation of the product. At the same time, moisturizing ingredients such as hyaluronic acid and glycerol can be produced in a series of enzymolysis and fermentation processes. Through the joint action of multiple strains, the output of these moisturizing ingredients can be increased, thereby improving the moisturizing performance of the product.

[0022] Furthermore, in step (4), 0.20-0.45 μm filter membrane is used for filtration.

[0023] Furthermore, the concentration of the ethanol aqueous solution in step (4) is 70-80 (v / v)%.

[0024] Furthermore, in step (4), an equal volume of ethanol aqueous solution is added to the fermentation product.

[0025] Furthermore, in step (4), the supernatant is concentrated by rotary evaporation using a vacuum rotary evaporator to 1 / 17-1 / 15 of the volume of the supernatant.

[0026] Compared with the prior art, the advantages and beneficial effects of the present invention are:

[0027] 1. The present invention performs mixed enzymolysis and fermentation on rose petals, perilla roots, grape seeds and Manuka honey in a specific ratio, which can improve the antibacterial and anti-inflammatory stability of the product.

[0028] 2. The present invention can improve the irritation of the product by first using three enzymes for gradient enzymolysis and then using three bacteria for gradient fermentation. At the same time, the moisturizing performance of the product is also improved under this condition. DETAILED DESCRIPTION

[0029] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0030] The raw materials used in the following examples of the present invention are all commercially available products:

[0031] Pectinase, cellulase, neutral protease, ficin, β-glucanase, and bromelain were purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0032] Lactobacillus bulgaricus, number: CCTCCCB20082295, was purchased from China Center for Type Culture Collection.

[0033] Lactobacillus brevis was purchased from Shanghai Collection of Microorganisms, with the number: SHBCCD14346.

[0034] Lactobacillus casei was purchased from Shanghai Collection of Microorganisms Center, with the number: SHBCCD24737.

[0035] Lactobacillus acidophilus was purchased from China Center for Type Culture Collection, with the number: CCTCCAB2010208.

[0036] Lactobacillus helveticus was purchased from China Center for Type Culture Collection, with the number: CCTCCAB2010205.

[0037] Bifidobacterium lactis was purchased from Shanghai Microbiological Collection Center, with the number: SHBCCD24728.

[0038] Example 1

[0039] This embodiment provides an application of Manuka honey in skin antibacterial and anti-inflammatory treatment, and the application comprises the following steps:

[0040] (1) Rose petals, perilla roots, and grape seeds are mixed and crushed to a particle size of less than 100 mesh to obtain a powder, and the powder is pretreated. The steps of pretreating the powder are as follows: first, the powder is placed at a low temperature of -30°C for 3 minutes, then the temperature is rapidly raised to 30°C within 2 seconds, and the process is repeated twice; then the powder is pressurized to 17 MPa and maintained for 4 minutes, and then the pressure is released to normal pressure, and the process is repeated three times to obtain a pretreated powder. The pretreated powder is mixed with Manuka honey and added to water to obtain an enzymatic substrate; the weight ratio of rose petals, perilla roots, grape seeds, and Manuka honey is 3:9:10:14; the weight of the added water is 12 times the total weight of the rose petals, perilla roots, grape seeds, and Manuka honey.

[0041] (2) adding pectinase to the enzymatic substrate, and performing enzymatic hydrolysis at 37°C for 70 minutes; inactivating the enzyme, adding neutral protease, and performing enzymatic hydrolysis at 37°C for 140 minutes; inactivating the enzyme, and continuously adding β-glucanase, and performing enzymatic hydrolysis at 37°C for 110 minutes; inactivating the enzyme to obtain an enzymatic hydrolysis product; the dosage of pectinase is 700u / mL enzymatic substrate; the dosage of neutral protease is 1100u / mL enzymatic substrate; the dosage of β-glucanase is 1500u / mL enzymatic substrate.

[0042] (3) Add Lactobacillus brevis to the enzymatic hydrolysate, ferment at 42° C. for 3.5 h, continue to add Lactobacillus helveticus, ferment at 42° C. for 6.5 h, and finally add Bifidobacterium lactis, ferment at 42° C. for 4.5 h, sterilize, and obtain a fermentation product; the amount of Lactobacillus brevis is 10 6 / mL enzymatic hydrolysate, the dosage of Lactobacillus helveticus is 10 9 / mL enzymatic hydrolysate, the dosage of Bifidobacterium lactis is 10 7 / mL enzymatic hydrolysis product.

[0043] (4) adding an equal volume of 75 (v / v)% ethanol aqueous solution to the fermentation product, stirring at 250 rpm / min for 1.5 h, filtering with a 0.22 μm filter membrane, and concentrating the obtained supernatant to 1 / 16 of the volume of the supernatant using a vacuum rotary evaporator to obtain a mixed extract;

[0044] (5) The mixed extract is used to prepare skin antibacterial and anti-inflammatory products.

[0045] Example 2

[0046] This embodiment provides an application of Manuka honey in skin antibacterial and anti-inflammatory treatment, and the application comprises the following steps:

[0047] (1) Rose petals, perilla roots, and grape seeds are mixed and crushed to a particle size of less than 100 mesh to obtain a powder, and the powder is pretreated. The steps of pretreating the powder are as follows: first, the powder is placed at a low temperature of -30°C for 2 minutes, then the temperature is rapidly raised to 30°C within 2 seconds, and the process is repeated twice; then the powder is pressurized to 16 MPa and maintained for 4 minutes, and then the pressure is released to normal pressure, and the process is repeated twice to obtain a pretreated powder, and the pretreated powder is mixed with Manuka honey and added to water to obtain an enzymatic substrate; the weight ratio of rose petals, perilla roots, grape seeds, and Manuka honey is 2:8:10:13; the weight of the added water is 10 times the total weight of the rose petals, perilla roots, grape seeds, and Manuka honey.

[0048] (2) adding pectinase to the enzymatic substrate, and performing enzymatic hydrolysis at 35°C for 60 minutes; inactivating the enzyme, adding neutral protease, and performing enzymatic hydrolysis at 35°C for 130 minutes; inactivating the enzyme, and continuously adding β-glucanase, and performing enzymatic hydrolysis at 35°C for 100 minutes; inactivating the enzyme, and obtaining an enzymatic hydrolysis product; the dosage of pectinase is 600u / mL enzymatic substrate; the dosage of neutral protease is 1000u / mL enzymatic substrate; the dosage of β-glucanase is 1400u / mL enzymatic substrate.

[0049] (3) Add Lactobacillus brevis to the enzymatic hydrolysate, ferment at 40°C for 3 h, continue to add Lactobacillus helveticus, ferment at 40°C for 6 h, and finally add Bifidobacterium lactis, ferment at 40°C for 4 h, sterilize, and obtain a fermentation product; the amount of Lactobacillus brevis is 10 6 / mL enzymatic hydrolysate, the dosage of Lactobacillus helveticus is 10 8 / mL enzymatic hydrolysate, the dosage of Bifidobacterium lactis is 10 7 / mL enzymatic hydrolysis product.

[0050] (4) adding an equal volume of 70 (v / v)% ethanol aqueous solution to the fermentation product, stirring at 200 rpm / min for 1 h, filtering with a 0.20 μm filter membrane, and concentrating the obtained supernatant to 1 / 17 of the volume of the supernatant using a vacuum rotary evaporator to obtain a mixed extract;

[0051] (5) The mixed extract is used to prepare skin antibacterial and anti-inflammatory products.

[0052] Example 3

[0053] This embodiment provides an application of Manuka honey in skin antibacterial and anti-inflammatory treatment, and the application comprises the following steps:

[0054] (1) Rose petals, perilla roots, and grape seeds are mixed and crushed to a particle size of less than 100 mesh to obtain a powder, and the powder is pretreated. The steps of pretreating the powder are as follows: first, the powder is placed at a low temperature of -30°C for 3 minutes, then the temperature is rapidly raised to 30°C within 2 seconds, and the process is repeated 3 times; then the powder is pressurized to 18 MPa and maintained for 5 minutes, and then the pressure is released to normal pressure, and the process is repeated 3 times to obtain a pretreated powder, and the pretreated powder is mixed with Manuka honey and added to water to obtain an enzymatic substrate; the weight ratio of rose petals, perilla roots, grape seeds, and Manuka honey is 4:10:10:15; the weight of the added water is 15 times the total weight of the rose petals, perilla roots, grape seeds, and Manuka honey.

[0055] (2) adding pectinase to the enzymatic substrate, and performing enzymatic hydrolysis at 38°C for 80 minutes; inactivating the enzyme, adding neutral protease, and performing enzymatic hydrolysis at 38°C for 150 minutes; inactivating the enzyme, and continuously adding β-glucanase, and performing enzymatic hydrolysis at 38°C for 120 minutes; inactivating the enzyme, and obtaining an enzymatic hydrolysis product; the dosage of pectinase is 800u / mL of enzymatic substrate; the dosage of neutral protease is 1200u / mL of enzymatic substrate; the dosage of β-glucanase is 1600u / mL of enzymatic substrate.

[0056] (3) Add Lactobacillus brevis to the enzymatic hydrolysate, ferment at 45°C for 4 hours, continue to add Lactobacillus helveticus, ferment at 45°C for 7 hours, and finally add Bifidobacterium lactis, ferment at 45°C for 5 hours, sterilize, and obtain a fermentation product; the amount of Lactobacillus brevis is 10 7 / mL enzymatic hydrolysate, the dosage of Lactobacillus helveticus is 10 9 / mL enzymatic hydrolysate, the dosage of Bifidobacterium lactis is 10 8 / mL enzymatic hydrolysis product.

[0057] (4) adding an equal volume of 80 (v / v)% ethanol aqueous solution to the fermentation product, stirring at 300 rpm / min for 2 h, filtering with a 0.45 μm filter membrane, and concentrating the obtained supernatant to 1 / 15 of the volume of the supernatant using a vacuum rotary evaporator to obtain a mixed extract;

[0058] (5) The mixed extract is used to prepare skin antibacterial and anti-inflammatory products.

[0059] Comparative Example 1

[0060] The difference between this comparative example and Example 1 is that the weight ratio of rose petals, perilla roots, grape seeds and Manuka honey is 1:1:1:1.

[0061] Comparative Example 2

[0062] The difference between this comparative example and Example 1 is that the powder is not pretreated.

[0063] Comparative Example 3

[0064] The difference between this comparative example and Example 1 is that rose petals, perilla roots and grape seeds are replaced with green tea, Poria cocos and Astragalus membranaceus in sequence.

[0065] Comparative Example 4

[0066] The difference between this comparative example and Example 1 is that pectinase is replaced by cellulase; neutral protease is replaced by ficin; β-glucanase is replaced by bromelain.

[0067] Comparative Example 5

[0068] The difference between this comparative example and Example 1 is that the first two enzymes in step (2) are not inactivated after the enzymatic hydrolysis is completed. Specifically: (2) pectinase is added to the enzymatic substrate, and the enzymatic hydrolysis is carried out at 37°C for 70 minutes; neutral protease is added, and the enzymatic hydrolysis is carried out at 37°C for 140 minutes; β-glucanase is continuously added, and the enzymatic hydrolysis is carried out at 37°C for 110 minutes; the enzyme is inactivated to obtain an enzymatic hydrolysis product; the amount of pectinase is 700u / mL enzymatic substrate; the amount of neutral protease is 1100u / mL enzymatic substrate; the amount of β-glucanase is 1500u / mL enzymatic substrate.

[0069] Comparative Example 6

[0070] The difference between this comparative example and Example 1 is that Lactobacillus brevis is replaced by Lactobacillus acidophilus; Lactobacillus helveticus is replaced by Lactobacillus bulgaricus, and Bifidobacterium lactis is replaced by Lactobacillus casei.

[0071] Comparative Example 7

[0072] The difference between this comparative example and Example 1 is that in step (3), after the first two bacteria are fermented, they are sterilized. Specifically, Lactobacillus brevis is added to the enzymatic hydrolyzate, fermented at 42°C for 3.5 hours, sterilized, then Lactobacillus helveticus is added, fermented at 42°C for 6.5 hours, sterilized, and finally Bifidobacterium lactis is added, fermented at 42°C for 4.5 hours, sterilized, and a fermentation product is obtained; the amount of Lactobacillus brevis is 10 6 / mL enzymatic hydrolysate, the dosage of Lactobacillus helveticus is 10 9 / mL enzymatic hydrolysate, the dosage of Bifidobacterium lactis is 10 7 / mL enzymatic hydrolysis product.

[0073] Comparative Example 8

[0074] The difference between this comparative example and Example 1 is that only Manuka honey is used.

[0075] An application of Manuka honey in skin antibacterial and anti-inflammatory treatment, the application comprising the following steps:

[0076] (1) Adding Manuka honey to water to obtain an enzymatic substrate; the weight of the added water is 12 times the total weight of the Manuka honey.

[0077] (2) adding pectinase to the enzymatic substrate, and performing enzymatic hydrolysis at 37°C for 70 minutes; inactivating the enzyme, adding neutral protease, and performing enzymatic hydrolysis at 37°C for 140 minutes; inactivating the enzyme, and continuously adding β-glucanase, and performing enzymatic hydrolysis at 37°C for 110 minutes; inactivating the enzyme to obtain an enzymatic hydrolysis product; the dosage of pectinase is 700u / mL enzymatic substrate; the dosage of neutral protease is 1100u / mL enzymatic substrate; the dosage of β-glucanase is 1500u / mL enzymatic substrate.

[0078] (3) Add Lactobacillus brevis to the enzymatic hydrolysate, ferment at 42° C. for 3.5 h, continue to add Lactobacillus helveticus, ferment at 42° C. for 6.5 h, and finally add Bifidobacterium lactis, ferment at 42° C. for 4.5 h, sterilize, and obtain a fermentation product; the amount of Lactobacillus brevis is 10 6 / mL enzymatic hydrolysate, the dosage of Lactobacillus helveticus is 10 9 / mL enzymatic hydrolysate, the dosage of Bifidobacterium lactis is 10 7 / mL enzymatic hydrolysis product.

[0079] (4) adding an equal volume of 75 (v / v)% ethanol aqueous solution to the fermentation product, stirring at 250 rpm / min for 1.5 h, filtering with a 0.22 μm filter membrane, and concentrating the obtained supernatant to 1 / 16 of the volume of the supernatant using a vacuum rotary evaporator to obtain a mixed extract;

[0080] (5) The mixed extract is used to prepare skin antibacterial and anti-inflammatory products.

[0081] Performance Testing

[0082] The mixed extracts prepared in Examples 1-3 and Comparative Examples 1-8 were subjected to performance tests.

[0083] 1. Human skin patch test

[0084] Test sample: mixed extracts prepared in Examples 1-3 and Comparative Examples 1-8;

[0085] Test population: Volunteers with sensitive skin aged 20-50 years old diagnosed in the dermatology department of a tertiary hospital were selected and randomly divided into 10 groups, with 15 people in each group;

[0086] Test method: First clean the skin of the test area with physiological saline and wait for 5 minutes to dry; select a qualified patch tester, use a closed patch test method, take 0.5mL of the mixed extract in the patch tester, apply it to the back of the subject with special tape, and ask the subject how it feels; bathing is prohibited during the test; apply the cream sample, and the subject should keep the patch area dry within 48 hours, avoid intense exercise, scratching the patch area, long-term sunlight exposure, etc. After 48 hours, remove the tester and mark it. After 30 minutes, wait until the indentation disappears and make a judgment under sufficient light. Record the results according to the skin reaction grade standard in the "Hygiene Standards for Skin Care Products". The skin adverse reaction grade standard is shown in Table 1 below. If there is any discomfort during the process, stop the test immediately. The results are shown in Table 2.

[0087] Table 1 Grading of skin adverse reactions

[0088]

[0089] Table 2 Human skin patch test results

[0090]

[0091] 2. Antibacterial effect test

[0092] The test was conducted in accordance with the microbial challenge test recommended by the International Cosmetic, Fragrance and Detergent Association (CTFA). The mixed extracts of Examples 1-3 and Comparative Examples 1-8 were weighed and added with bacteria in the first, third and fifth weeks of the test, respectively. The amount of bacteria added each time was 1×10 6 CFU / mL, the bacterial content in the sample was analyzed on the 14th day after the addition of bacteria. The types of bacteria are: Staphylococcus aureus (ATCC6538), Escherichia coli (ATCC25922), Candida albicans (ATCC14053), Pseudomonas aeruginosa (ATCC27853), Aspergillus niger (ATCC16404), Bacillus subtilis (ATCC6633). Each bacterium was tested separately.

[0093] Evaluation method:

[0094] (1) The antiseptic effect is excellent, that is, after adding bacteria three times, at the 14th hour after each addition, the amount of surviving bacteria is reduced to no more than 0.01% of the initial concentration, and the antiseptic effect of the sample is excellent.

[0095] (2) The preservative effect is acceptable, that is, after three times of adding bacteria, at the 14th hour after each addition of bacteria, the amount of surviving bacteria is reduced to no more than 0.1% of the initial concentration, and the sample passes the test.

[0096] (3) The antiseptic effect is poor, that is, after adding bacteria three times, at the 14th hour after each addition, the surviving bacteria is greater than 0.1%, and the sample fails the test.

[0097] The results are shown in Table 3.

[0098] Table 3 Antibacterial test results

[0099]

[0100]

[0101]

[0102]

[0103] 3. Moisturizing effect

[0104] Test population: Female volunteers aged 20-50 years old with healthy skin were selected.

[0105] Test instrument: Skin moisture content tester (Cormeter CM825; CK, Germany): uses the capacitance method to measure the moisture content of the stratum corneum of human skin.

[0106] Test steps:

[0107] 55 volunteers were randomly selected and divided into 11 groups. After cleansing, the mixed extracts were applied to the volunteers' faces. No washing was required and the mixture was absorbed naturally. The mixture was applied once every night for 4 consecutive weeks.

[0108] Before the test, each group of volunteers cleaned their face with clean water, wiped their face, and sat quietly for 30 minutes in a constant temperature and humidity laboratory with a temperature of 20±1℃ and a humidity of 50±2% before starting the test. A 4*4cm area in the middle of the left face of each volunteer was selected, and the probe was used to test the average capacitance value of the four corners and a total of 5 positions in the middle of the area. Then the capacitance values ​​of the volunteers before and after the experiment were tested respectively, and the difference between the average capacitance values ​​of each group of people before and after the experiment was recorded.

[0109] Evaluation method: The MMV value represents skin moisture. The greater the difference in MMV before and after the experiment, the better the moisturizing effect of the mixed extract.

[0110] The experimental results are shown in Table 4.

[0111] Table 4 Performance test results

[0112] MMV Difference Example 1 8.7 Example 2 8.3 Example 3 8.4 Comparative Example 1 7.8 Comparative Example 2 8.0 Comparative Example 3 7.5 Comparative Example 4 5.9 Comparative Example 5 6.7 Comparative Example 6 5.8 Comparative Example 7 6.5 Comparative Example 8 3.2

[0113] From the above performance test results, it can be seen that the mixed extracts of Examples 1-3 have good antibacterial and anti-inflammatory effects, high stability, and good moisturizing effects, especially the comprehensive performance of Example 1 is the most outstanding.

[0114] The comparative examples, however, were significantly inferior to the embodiments in terms of the corresponding performance tests because they did not adopt the necessary technical solutions. In comparative example 1, the ratio of rose petals, perilla roots, grape seeds and Manuka honey was changed, and the antibacterial stability of the prepared mixed extract decreased, indicating that only when rose petals, perilla roots, grape seeds and Manuka honey were enzymatically fermented at a specific ratio, a higher antibacterial stability of the product could be obtained. The powder of comparative example 2 was not pretreated, and the active components could not be extracted into the product to the greatest extent, affecting the antibacterial stability of the product. In comparative example 3, rose petals, perilla roots and grape seeds were replaced with green tea, Poria cocos and Astragalus membranaceus in sequence, resulting in different compositions of the effective components of the product, which reduced the antibacterial stability. Comparative examples 4 and 6 used different types of enzymes and bacteria, respectively, and the irritation of the product became greater, and the moisturizing effect decreased. In comparative example 5, the first two enzymes in step (2) were not inactivated after the enzymolysis was completed, which affected the moisturizing effect of the product, indicating that in the enzymolysis stage, the previous enzymes and the enzymes added later were mixed, but played an inhibitory role. In comparative example 7, after the first two bacteria were fermented in step (3), they were sterilized, and the moisturizing effect decreased, indicating that the first bacteria and the latter bacteria can work together. In comparative example 8, only honey was used for fermentation, and the irritation of the obtained mixed extract was significantly increased, the antibacterial stability was poor, and the moisturizing effect decreased. The above experimental results further prove the importance of the technical solution defined in the present invention for its technical effect.

[0115] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. An application of Manuka honey in skin antibacterial and anti-inflammatory treatment, characterized in that: The application comprises the following steps: (1) mixing rose petals, perilla roots, and grape seeds, and crushing them to a particle size of less than 100 mesh to obtain a powder, pre-treating the powder to obtain a pre-treated powder, mixing the pre-treated powder with Manuka honey, and adding the mixture to water to obtain an enzymatic substrate; (2) adding pectinase to the enzymatic substrate, performing enzymatic hydrolysis at 35° C.-38° C. for 60-80 min; inactivating the enzyme, adding neutral protease, performing enzymatic hydrolysis at 35° C.-38° C. for 130-150 min; inactivating the enzyme, continuing to add β-glucanase, performing enzymatic hydrolysis at 35° C.-38° C. for 100-120 min; inactivating the enzyme, and obtaining an enzymatic hydrolysis product; (3) adding Lactobacillus brevis to the enzymatic hydrolysate, fermenting at 40° C.-45° C. for 3-4 h, continuing to add Lactobacillus helveticus, fermenting at 40° C.-45° C. for 6-7 h, and finally adding Bifidobacterium lactis, fermenting at 40° C.-45° C. for 4-5 h, sterilizing, and obtaining a fermentation product; (4) adding ethanol aqueous solution to the fermentation product, stirring for 1-2 hours, filtering with a filter membrane to obtain a supernatant, and concentrating the supernatant to obtain a mixed extract; (5) The mixed extract is used to prepare skin antibacterial and anti-inflammatory products.

2. The use of Manuka honey in skin antibacterial and anti-inflammatory treatment according to claim 1, characterized in that: In the step (1), the weight ratio of rose petals, perilla roots, grape seeds and Manuka honey is (2-4): (8-10): 10: (13-15).

3. The use of Manuka honey in skin antibacterial and anti-inflammatory treatment according to claim 2, characterized in that: The weight of water added in step (1) is 10-15 times the total weight of rose petals, perilla roots, grape seeds and Manuka honey.

4. The use of Manuka honey in skin antibacterial and anti-inflammatory treatment according to claim 1, characterized in that: The steps of pre-treating the powder in step (1) are as follows: firstly, the powder is placed in a low temperature of -30°C for 2-3 minutes, then the temperature is rapidly raised to 30°C within 2 seconds, and the process is repeated 2-3 times; then the powder is pressurized to 16-18MPa, maintained for 4-5 minutes, and then the pressure is released to normal pressure, and the process is repeated 2-3 times to obtain the pre-treated powder.

5. The use of Manuka honey in skin antibacterial and anti-inflammatory treatment according to claim 1, characterized in that: In the step (2), the dosage of pectinase is 600-800u / mL of enzymatic substrate; the dosage of neutral protease is 1000-1200u / mL of enzymatic substrate; and the dosage of β-glucanase is 1400-1600u / mL of enzymatic substrate.

6. The use of Manuka honey in skin antibacterial and anti-inflammatory treatment according to claim 5, characterized in that: The amount of Lactobacillus brevis in step (3) is 10 6 -10 7 / mL enzymatic hydrolysate, the dosage of Lactobacillus helveticus is 10 8 -10 9 / mL enzymatic hydrolysate, the dosage of Bifidobacterium lactis is 10 7 -10 8 / mL enzymatic hydrolysis product.

7. The use of Manuka honey in skin antibacterial and anti-inflammatory treatment according to claim 1, characterized in that: In the step (4), a 0.20-0.45 μm filter membrane is used for filtration.

8. The use of Manuka honey in skin antibacterial and anti-inflammatory treatment according to claim 1, characterized in that: The concentration of the ethanol aqueous solution in step (4) is 70-80 (v / v)%.

9. The use of Manuka honey in skin antibacterial and anti-inflammatory treatment according to claim 8, characterized in that: In the step (4), an equal volume of ethanol aqueous solution is added to the fermentation product.

10. The use of Manuka honey in skin antibacterial and anti-inflammatory treatment according to claim 1, characterized in that: In the step (4), the supernatant is concentrated by rotary evaporation using a vacuum rotary evaporator to 1 / 17-1 / 15 of the volume of the supernatant.

Images