A mixed oil for repairing sensitive skin, and a preparation method and application thereof

Abstract

The application provides a mixed oil for repairing sensitive skin and a preparation method and application thereof, and raw materials of the mixed oil include peony seed, Prinsepia uniflora fruit and camellia seed. The mixed oil prepared by the application has excellent anti-inflammatory, moisturizing and barrier repairing effects, and the peony seed, Prinsepia uniflora fruit and camellia seed have certain mutual cooperation and synergistic effect on the above effects, and a specific extraction method can promote the maximization of the effect of the oil.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of cosmetics, and particularly relates to a mixed oil for repairing sensitive skin and a preparation method and application thereof. BACKGROUND

[0002] Modern fast-paced lifestyle has brought many adverse effects on the skin. Long-term mental stress can cause endocrine disorders in the human body, affecting the sebaceous gland secretion and blood circulation of the skin. Imbalance of sebaceous gland secretion can make the skin too oily or dry, and poor blood circulation can lead to insufficient nutrient supply to the skin, resulting in dull and lackluster skin, and reducing the self-repairing ability of the skin.

[0003] Sensitive skin refers to a high reaction state of the skin under physiological or pathological conditions, which can exhibit subjective symptoms such as burning, stinging, itching and tightness, and objective signs such as erythema, telangiectasia and desquamation. As the largest organ of the human body, the skin has important functions such as protecting the body from external harm, regulating body temperature, and perceiving external stimuli. Maintaining the healthy state of the skin, especially its soothing and moisturizing performance, is crucial for maintaining the normal physiological functions and overall health of the skin. Soothing and moisturizing skin can effectively resist external stimuli, reduce the risk of skin diseases, and at the same time improve the appearance and texture of the skin, making people look younger, healthier and more energetic. With the continuous improvement of consumers' awareness of skin care, their demand for skin care products that can simultaneously meet the soothing and moisturizing dual functions, and are safe, effective and long-lasting, is increasingly urgent. This market demand provides strong impetus for the development of new soothing and moisturizing compositions, prompting researchers to continuously explore and innovate to develop better skin care products to meet the growing needs of consumers.

[0004] In view of the various adverse effects of environmental factors and lifestyle on the skin and the limitations of existing skin care products in soothing and moisturizing, it is of great significance and broad market prospect to develop a soothing and moisturizing composition that is efficient, stable and safe. SUMMARY

[0005] In view of the deficiencies of the prior art, the purpose of the present application is to provide a mixed oil for repairing sensitive skin and a preparation method and application thereof.

[0006] To achieve the purpose of the present application, the following technical solutions are adopted:

[0007] In a first aspect, the present application provides a mixed oil for repairing sensitive skin, wherein the preparation raw materials of the mixed oil comprise Yunnan peony seeds, Chinese olive fruits and camellia seeds.

[0008] The Dian-mandarin seed is rich in unsaturated fatty acids such as linolenic acid and linoleic acid. Linolenic acid has the effects of lowering blood lipids, lowering cholesterol, promoting fat metabolism, liver cell regeneration, immunity, anti-allergic reaction, delaying aging, etc. Linoleic acid has the effects of inhibiting the synthesis of cholesterol in the human body, antioxidation, anticancer, and preventing diabetes. These components help maintain the healthy state of the skin, enhance the immune resistance of the skin, and reduce the damage of the external environment to the skin. In addition, the Dian-mandarin seed is also rich in anthocyanins and other components. Such components have strong anti-inflammatory, barrier repair and moisturizing effects on the skin.

[0009] The Qing-stone fruit is high in oil content, and is rich in unsaturated fatty acids, polyphenols, flavones and other components. These components can form a protective film on the surface of the skin, effectively preventing the loss of water in the skin. This protective film acts as a "barrier", reducing the contact of the skin with the dry environment, thereby achieving good barrier skin care effect.

[0010] The Camellia seed mainly contains fatty acids, tea seed polysaccharides, polyphenols, flavones, tea saponin and other components. Among them, fatty acids, tea polyphenols and camellianin are characteristic bioactive substances, which have the effects of lowering cholesterol, anti-aging and moisturizing.

[0011] The mixed oil prepared by the present application has excellent anti-inflammatory, moisturizing and barrier repair effects, and the Dian-mandarin seed, Qing-stone fruit and Camellia seed have certain mutual synergistic effects on the above effects.

[0012] Preferably, the preparation raw materials of the mixed oil include 4-6 parts of Dian-mandarin seed, 1-3 parts of Qing-stone fruit and 1-2 parts of Camellia seed by mass fraction.

[0013] The mass fraction of the Dian-mandarin seed can be selected from 4 parts, 4.2 parts, 4.4 parts, 4.6 parts, 4.8 parts, 5 parts, 5.2 parts, 5.4 parts, 5.6 parts, 5.8 parts, 6 parts, etc. The mass fraction of the Qing-stone fruit can be selected from 1 part, 1.2 parts, 1.4 parts, 1.6 parts, 1.8 parts, 2 parts, 2.2 parts, 2.4 parts, 2.6 parts, 2.8 parts, 3 parts, etc. The mass fraction of the Camellia seed can be selected from 1 part, 1.1 part, 1.2 part, 1.3 part, 1.4 part, 1.5 part, 1.6 part, 1.7 part, 1.8 part, 1.9 part, 2 part, etc. Other specific point values in the above numerical range can be selected, which will not be described here.

[0014] In the second aspect, the present application provides a preparation method of the mixed oil for repairing sensitive skin according to the first aspect, which comprises:

[0015] (1) mixing the preparation raw materials with the fermentation strain, fermenting, drying to obtain a fermented mixture;

[0016] (2) the fermentation mixture is extracted by subcritical extraction to obtain crude oil;

[0017] (3) the crude oil is filtered to obtain the product.

[0018] The specific extraction method of the present application can not only extract fatty acid components in the mixed raw materials, but also extract small molecules such as polyphenols, flavonoids, and anthocyanins in the seed coat of the raw materials. These components can promote the maximization of the effect in cooperation with each other. The mixed oil obtained by the method of mixing the raw materials and then fermenting is better than the mixed oil obtained by the method of separately extracting the extract and then physically mixing. The extraction method can make the molecular connection between different oils more closely, and the mixed oil obtained by the preparation method has better anti-inflammatory, moisturizing, and barrier repair effects than the oil obtained by traditional methods such as pressing and supercritical CO2 extraction.

[0019] Preferably, before the preparation raw materials are mixed with the fermentation strain, the preparation raw materials are also mixed, crushed, and sieved.

[0020] Preferably, the fermentation strain includes Aspergillus niger and Bacillus pumilus with a ratio of viable bacterial count of 1: (1-3).

[0021] The specific point values in (1-3) can be selected from 1, 1.2, 1.4, 1.6, 1.8, 2, 2.2, 2.4, 2.6, 2.8, 3, and other specific point values within the above numerical range can be selected, which will not be repeated here.

[0022] The use of the above specific ratio of mixed strains for fermentation treatment improves the extraction rate of the mixed oil, reduces the content of stearic acid and palmitic acid in the saturated fatty acids, and increases the content of linolenic acid in the polyunsaturated fatty acids. This changes the composition ratio of fatty acids in the mixed oil and increases the content of small molecule effective substances in the mixed oil, thereby resulting in better soothing, moisturizing, and barrier repair effects.

[0023] Preferably, the addition amount of the fermentation strain is 2x10 8 CFU / g-5x10 8 CFU / g, for example, 2x10 8 CFU / g, 2.2x10 8 CFU / g, 2.5x10 8 CFU / g, 2.8x10 8 CFU / g, 3x10 8 CFU / g, 3.2x10 8 CFU / g, 3.5x10 8 CFU / g, 3.8x10 8 CFU / g, 4x10 8 CFU / g, 4.2x108 CFU / g, 4.5 x 10 8 CFU / g, 4.8 x 10 8 CFU / g, 5 x 10 8 CFU / g, etc. Other specific point values within the above numerical ranges can be selected, and will not be repeated here.

[0024] Preferably, the fermentation temperature is 30-40℃, the fermentation time is 48-96h, and the moisture content of the fermentation material is 30-50%.

[0025] The temperature can be selected as 30℃, 31℃, 32℃, 33℃, 34℃, 35℃, 36℃, 37℃, 38℃, 39℃, 40℃, etc., the time can be selected as 48h, 52h, 56h, 60h, 64h, 68h, 72h, 76h, 80h, 84h, 88h, 92h, 96h, etc., and the moisture content can be selected as 30%, 32%, 34%, 36%, 38%, 40%, 42%, 44%, 46%, 48%, 50%, etc. Other specific point values within the above numerical ranges can be selected, and will not be repeated here.

[0026] Preferably, the extraction solvent used in the subcritical extraction is anhydrous ethanol and butane in a volume ratio of 1:(4-7).

[0027] The specific point values in (4-7) can be selected as 4, 4.2, 4.5, 4.8, 5, 5.2, 5.5, 5.8, 6, 6.2, 6.5, 6.8, 7, etc. Other specific point values within the above numerical ranges can be selected, and will not be repeated here.

[0028] Preferably, the ratio of the fermentation mixture to the extraction solvent is 1g:(2-10)mL.

[0029] The specific point values in (2-10) can be selected as 2, 3, 4, 5, 6, 7, 8, 9, 10, etc. Other specific point values within the above numerical ranges can be selected, and will not be repeated here.

[0030] Preferably, the temperature of the subcritical extraction is 35-45℃, and the single extraction time is 2-3h.

[0031] The temperature can be selected as 35℃, 36℃, 37℃, 38℃, 39℃, 40℃, 41℃, 42℃, 43℃, 44℃, 45℃, etc., and the time can be selected as 2h, 2.1h, 2.2h, 2.3h, 2.4h, 2.5h, 2.6h, 2.7h, 2.8h, 2.9h, 3h, etc. Other specific point values within the above numerical ranges can be selected, and will not be repeated here.

[0032] Preferably, the subcritical extraction is performed at a pressure of 0.3-0.5 MPa and for 2-3 times.

[0033] The pressure can be selected from 0.3 MPa, 0.32 MPa, 0.34 MPa, 0.36 MPa, 0.38 MPa, 0.4 MPa, 0.42 MPa, 0.44 MPa, 0.46 MPa, 0.48 MPa, 0.5 MPa, and the like, and other specific point values within the above numerical range can be selected, which will not be repeated here.

[0034] In a third aspect, the present application provides a use of the mixed oil for repairing sensitive skin according to the first aspect in the preparation of a product having soothing, moisturizing or barrier repair effects.

[0035] Preferably, the mixed oil for repairing sensitive skin in the product has a mass percentage content of 0.1-0.5%, such as 0.1%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.18%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, and the like, and other specific point values within the above numerical range can be selected, which will not be repeated here.

[0036] In a fourth aspect, the present application provides a use of the mixed oil for repairing sensitive skin according to the first aspect in the preparation of an inflammation factor inhibitor, an AQP3 gene activator, an LOR gene activator or an FLG gene activator.

[0037] Preferably, the inflammation factor includes TNF-α, IL-6 or NO.

[0038] According to the research results of the present application, the mixed oil prepared by the present application has a significant inhibitory effect on the secretion of inflammation factors, and has an activating effect on the AQP3 gene, the LOR gene and the FLG gene, so that the prepared oil can be used as an in vitro preparation for scientific basic research, such as exploring the mechanism of physiological activities related to inflammatory response, or for the expression of barrier activating genes.

[0039] Compared with the prior art, the present application has the following beneficial effects:

[0040] The mixed oil prepared by the method has more excellent anti-inflammatory, moisturizing and barrier repair effects, and the Yunnan peony seeds, Chinese pricklyash fruits and camellia seeds have certain mutual cooperation and synergistic effect on the above effects. The specific extraction method can promote the effect maximization. The oil obtained by the preparation method has higher yield than the oil obtained by the traditional oil preparation method, retains part of the small molecule effective substances (such as flavonoids, anthocyanins and the like), and has higher unsaturated fatty acid content, so that the composition ratio of each fatty acid is changed, and the oil has more excellent anti-inflammatory, moisturizing and barrier repair effects. DETAILED DESCRIPTION

[0041] The technical solutions of the present application are further illustrated by specific embodiments. Those skilled in the art should understand that the embodiments are only used to help understand the present application and should not be regarded as specific limitations on the present application.

[0042] The effective components in the products involved in the following examples and comparative examples are as follows (only the effective components are embodied, and the necessary auxiliary components contained in other commercially available raw materials are not described again) :

[0043] The preservation number of the Aspergillus niger is CGMCC No.3.3928, which is purchased from the Beijing Institute of Microbiology and Epidemiology.

[0044] The preservation number of the Bacillus pumilus is CMCC(B)63202, which is purchased from the Beijing Institute of Microbiology and Epidemiology.

[0045] Example 1

[0046] The present embodiment provides a mixed oil for repairing sensitive skin, and the preparation raw materials of the mixed oil include 5 parts of Yunnan peony seeds, 2 parts of Chinese pricklyash fruits and 1.5 parts of camellia seeds in terms of mass fraction.

[0047] The preparation method is as follows:

[0048] (1) The preparation raw materials are mixed and crushed, and then passed through a 60-mesh sieve for standby. Aspergillus niger (CGMCC No.3.3928) and Bacillus pumilus (CMCC(B)63202) with a viable bacterial count ratio of 1:2 are inoculated into the crushed material with a water content of 40%, and the inoculation amount is 3.5×10 8 CFU / g. After 60h of solid-state fermentation at 37℃, drying treatment is performed to obtain a first mixture.

[0049] (2) The first mixture is extracted by a subcritical extraction device, the ratio of the first mixture to the extraction solvent is 1g:6mL, the extraction solvent is anhydrous ethanol and butane with a volume ratio of 1:5, the extraction temperature is 40℃, the pressure is 0.4Mpa, the single extraction time is 2h, and the extraction times are 2 times to obtain a crude oil.

[0050] (3) The crude oil is filtered with a filter membrane of 0.45 μm, and the mixture oil is obtained.

[0051] Example 2

[0052] The mixture oil for repairing sensitive skin is prepared from 4 parts of Yunnan peony seeds, 3 parts of wild jujube fruits and 1 part of camellia seeds.

[0053] The preparation method is as follows:

[0054] (1) The raw materials are mixed and crushed, and then passed through a 60-mesh sieve for standby use. Aspergillus niger (CGMCC No. 3.3928) and Bacillus pumilus (CMCC(B)63202) with a viable bacterial count ratio of 1:1 are inoculated into the crushed material with a water content of 30%, and the inoculation amount is 2.5×10 8 CFU / g. After 96 h of solid-state fermentation at 30°C, drying treatment is performed to obtain a first mixture.

[0055] (2) The first mixture is extracted by a subcritical extraction device. The ratio of the first mixture to the extraction solvent is 1 g:3 mL. The extraction solvent is anhydrous ethanol and butane with a volume ratio of 1:4. The extraction temperature is 35°C, the pressure is 0.3 Mpa, the single extraction time is 2.5 h, and the extraction times are 3, to obtain crude oil.

[0056] (3) The crude oil is filtered with a filter membrane of 0.45 μm, and the mixture oil is obtained.

[0057] Example 3

[0058] The mixture oil for repairing sensitive skin is prepared from 4 parts of Yunnan peony seeds, 3 parts of wild jujube fruits and 1 part of camellia seeds.

[0059] The preparation method is as follows:

[0060] (1) The raw materials are mixed and crushed, and then passed through a 60-mesh sieve for standby use. Aspergillus niger (CGMCC No. 3.3928) and Bacillus pumilus (CMCC(B)63202) with a viable bacterial count ratio of 1:1 are inoculated into the crushed material with a water content of 30%, and the inoculation amount is 2.5×10 8 CFU / g. After 96 h of solid-state fermentation at 30°C, drying treatment is performed to obtain a first mixture.

[0061] (2) The first mixture is extracted by a subcritical extraction device. The ratio of the first mixture to the extraction solvent is 1 g:3 mL. The extraction solvent is anhydrous ethanol and butane with a volume ratio of 1:4. The extraction temperature is 35°C, the pressure is 0.3 Mpa, the single extraction time is 2.5 h, and the extraction times are 3, to obtain crude oil.

[0062] (3) The crude oil is filtered with a filter membrane of 0.45 μm, and the product is obtained.

[0063] Example 4

[0064] The present example provides a mixed oil for repairing sensitive skin, which is different from Example 1 only in that the step (2) of the preparation method is "the first mixture is pressed with a screw press device to obtain the crude oil".

[0065] The components and contents of the raw materials remain unchanged, and other preparation steps remain unchanged.

[0066] Example 5

[0067] The present example provides a mixed oil for repairing sensitive skin, which is different from Example 1 only in that the step (2) of the preparation method is "the first mixture is extracted with a supercritical CO2 extraction device, the CO2 flow rate is 20 L / h, the extraction temperature is 45℃, the pressure is 8 Mpa, the single extraction time is 2.5 h, and the number of times is 2, to obtain the crude oil".

[0068] The components and contents of the raw materials remain unchanged, and other preparation steps remain unchanged.

[0069] Example 6

[0070] The present example provides a mixed oil for repairing sensitive skin, which is different from Example 1 only in that the raw materials for preparing the mixed oil include 3 parts of Yunnan peony seeds, 4 parts of Chinese olive fruits, and 0.5 parts of camellia seeds by mass fraction.

[0071] The components and contents of the raw materials remain unchanged, and other preparation steps remain unchanged.

[0072] Example 7

[0073] The present example provides a mixed oil for repairing sensitive skin, which is different from Example 1 only in that the raw materials for preparing the mixed oil include 7 parts of Yunnan peony seeds, 0.5 parts of Chinese olive fruits, and 3 parts of camellia seeds by mass fraction.

[0074] The components and contents of the raw materials remain unchanged, and other preparation steps remain unchanged.

[0075] Example 8

[0076] The embodiment provides a mixed oil for repairing sensitive skin, which is different from the embodiment 1 only in that the step (2) is "extracting the first mixture by using a subcritical extraction device, the ratio of the first mixture to an extraction solvent is 1g:6ml, the extraction solvent is anhydrous ethanol and butane with a volume ratio of 1:8, the extraction temperature is 40 DEG C, the pressure is 0.4Mpa, the single extraction time is 2h, and the extraction times are 2, and the crude oil is obtained".

[0077] The components and contents of the raw materials remain unchanged, and other preparation steps remain unchanged.

[0078] Embodiment 9

[0079] The embodiment provides a mixed oil for repairing sensitive skin, which is different from the embodiment 1 only in that the step (2) is "extracting the first mixture by using a subcritical extraction device, the ratio of the first mixture to an extraction solvent is 1g:6ml, the extraction solvent is anhydrous ethanol and butane with a volume ratio of 1:3, the extraction temperature is 40 DEG C, the pressure is 0.4Mpa, the single extraction time is 2h, and the extraction times are 2, and the crude oil is obtained".

[0080] The components and contents of the raw materials remain unchanged, and other preparation steps remain unchanged.

[0081] Embodiment 10

[0082] The embodiment provides a mixed oil for repairing sensitive skin, which is different from the embodiment 1 only in that the step (1) is "pulverizing the raw materials, and then screening the pulverized materials through a 60-mesh screen, inoculating Aspergillus niger (CGMCC No.3.3928) into the pulverized materials with a water content of 40%, and the inoculation amount is 3.5x10 8 CFU / g, after 60h of solid state fermentation at 37 DEG C, drying treatment is carried out, and the first mixture is obtained".

[0083] The components and contents of the raw materials remain unchanged, and other preparation steps remain unchanged.

[0084] Embodiment 11

[0085] The embodiment provides a mixed oil for repairing sensitive skin, which is different from the embodiment 1 only in that the step (1) is "pulverizing the raw materials, and then screening the pulverized materials through a 60-mesh screen, inoculating Bacillus pumilus (CMCC(B)63202) into the pulverized materials with a water content of 40%, and the inoculation amount is 3.5x10 8 CFU / g, after 60h of solid state fermentation at 37 DEG C, drying treatment is carried out, and the first mixture is obtained".

[0086] The components and contents of the raw materials remain unchanged, and other preparation steps remain unchanged.

[0087] Example 12

[0088] The present example provides a mixed oil for repairing sensitive skin, which differs from Example 1 only in that step (1) is "pulverize the raw materials for preparation, reserve after passing through a 60-mesh sieve, and perform drying treatment after standing at 37°C for 60 h to obtain the first mixture".

[0089] The components and contents of the raw materials for preparation remain unchanged, and the other preparation steps remain unchanged.

[0090] Example 13

[0091] The present example provides a mixed oil for repairing sensitive skin, which differs from Example 1 only in that step (1) is "pulverize the raw materials for preparation, reserve after passing through a 60-mesh sieve, inoculate Aspergillus niger (CGMCC No. 3.3928) and Bacillus pumilus (CMCC(B) 63202) with a viable cell count ratio of 1:0.5 into the pulverized material with a water content of 40%, at an inoculation amount of 3.5 x 10 8 CFU / g, and perform drying treatment after solid-state fermentation at 37°C for 60 h to obtain the first mixture".

[0092] The components and contents of the raw materials for preparation remain unchanged, and the other preparation steps remain unchanged.

[0093] Example 14

[0094] The present example provides a mixed oil for repairing sensitive skin, which differs from Example 1 only in that step (1) is "pulverize the raw materials for preparation, reserve after passing through a 60-mesh sieve, inoculate Aspergillus niger (CGMCC No. 3.3928) and Bacillus pumilus (CMCC(B) 63202) with a viable cell count ratio of 1:5 into the pulverized material with a water content of 40%, at an inoculation amount of 3.5 x 10 8 CFU / g, and perform drying treatment after solid-state fermentation at 37°C for 60 h to obtain the first mixture".

[0095] The components and contents of the raw materials for preparation remain unchanged, and the other preparation steps remain unchanged.

[0096] Example 15

[0097] The present example provides a mixed oil for repairing sensitive skin, which differs from Example 1 only in that the preparation method is different.

[0098] The preparation method is as follows: separately prepare oils from Paeonia delavayi seeds, Prinsepia uniflora fruits, and Camellia sinensis seeds according to the preparation method of Example 1, and physically mix the Paeonia delavayi seed oil, Prinsepia uniflora fruit oil, and Camellia sinensis seed oil obtained to obtain the mixed oil.

[0099] Comparative Example 1

[0100] This comparative example provides a mixed oil for repairing sensitive skin, which is different from Example 1 only in that the raw material does not contain Malan seed, and the reduced mass is proportionally distributed to the mass of Prinsepia uniflora and Camellia seed, and other ingredients and contents remain unchanged.

[0101] The preparation method refers to Example 1.

[0102] Comparative Example 2

[0103] This comparative example provides a mixed oil for repairing sensitive skin, which is different from Example 1 only in that the raw material does not contain Prinsepia uniflora, and the reduced mass is proportionally distributed to the mass of Malan seed and Camellia seed, and other ingredients and contents remain unchanged.

[0104] The preparation method refers to Example 1.

[0105] Comparative Example 3

[0106] This comparative example provides a mixed oil for repairing sensitive skin, which is different from Example 1 only in that the raw material does not contain Camellia seed, and the reduced mass is proportionally distributed to the mass of Malan seed and Prinsepia uniflora, and other ingredients and contents remain unchanged.

[0107] The preparation method refers to Example 1.

[0108] Test Example 1

[0109] Anti-inflammatory soothing effect

[0110] LPS-induced mouse macrophage RAW264.7 cells were used as the anti-inflammatory efficacy test model. The blank control group, LPS induction group, positive control group (dexamethasone, 120 μg / mL) and sample group were set. The concentration of each example and comparative example sample was 120 μg / mL. After incubation at 37°C in a 5% CO2 incubator for 24 h, the cell supernatant of each group was collected, and then the content of inflammatory factors TNF-α, IL-6 and NO in the collected cell supernatant was detected according to the ELISA kit detection method. The experimental results were analyzed by data processing software to evaluate the inhibition rate of the mixed oil on the expression of inflammatory factors. The inhibition rate calculation formula is:

[0111]

[0112] Table 1

[0113]

[0114] From the data in Table 1, it can be seen that the mixed oil prepared by the application has good anti-inflammatory effect and can inhibit the expression of inflammatory factors; the peony seed, Chinese pricklyash fruit and camellia seed have certain mutual cooperation effect on the above-mentioned effect, and the three are indispensable; the pretreatment before fermentation can improve the technical effect of extraction; the common fermentation of aspergillus niger and bacillus pumilus can maximize the fermentation effect, and the two strains are indispensable; the effect of subcritical extraction is better than that of other extraction effects.

[0115] Test Example 2

[0116] Moisturizing effect

[0117] Logarithmic growth period, good HaCaT cells were selected and inoculated in a 24-well plate, 1×10 5 Cells / well, incubated in an incubator for 24h. Set up a blank control group, a positive control group (calcium chloride, 120 μg / mL) and a sample group, the concentration of the sample of the example and the comparative example was 120 μg / mL, the sample was treated with the cells, and the cells were incubated at 37℃ in a 5% CO2 incubator for 24h. Then extract the RNA of each group, determine the RNA content, reverse transcribe cDNA, fluorescence quantitative PCR experiment, analyze the experimental results by data processing software, and evaluate the activation effect of the mixed oil on the expression of water channel protein AQP3 mRNA. The activation rate calculation formula is:

[0118]

[0119] Table 2

[0120] AQP3 activation rate positive control group 16.23％ Example 1 74.85％ Example 2 73.59％ Example 3 72.64％ Example 4 41.32％ Example 5 43.16％ Example 6 36.85％ Example 7 40.56％ Example 8 47.21％ Example 9 49.03％ Example 10 45.10％ Example 11 41.48％ Example 12 34.93％ Example 13 50.31％ Example 14 47.83％ Example 15 49.60％ Comparative Example 1 36.84％ Comparative Example 2 32.85％ Comparative Example 3 21.05％

[0121] From the data in Table 2, it can be seen that the mixed oil prepared by the application has good moisturizing effect; the peony seed, Chinese pricklyash fruit and camellia seed have certain mutual cooperation effect on the above-mentioned effect, and the three are indispensable; the pretreatment before fermentation can improve the technical effect of extraction; the common fermentation of aspergillus niger and bacillus pumilus can maximize the fermentation effect, and the two strains are indispensable; the effect of subcritical extraction is better than that of other extraction effects.

[0122] Test Example 3

[0123] Barrier repair

[0124] Logarithmic growth period, good HaCaT cells were selected and inoculated in a 24-well plate, 1×10 5Cells / cone, incubated in the incubator for 24h. Set up control group and sample group, the concentration of sample group, example, comparative example sample is 120ug / mL, give sample processing cells, continue to incubate at 37℃, 5%CO2incubator for 24h. Extract RNA of each group, determine the RNA content, reverse transcription cDNA, fluorescence quantitative PCR experiment is analyzed by data processing software, evaluate the activation effect of mixed oil on barrier repair related filaggrin(FLG) and loricrin(LOR) mRNA expression. The activation rate is calculated by the following formula:

[0125]

[0126] Table 3

[0127]

[0128]

[0129] From the data in table 3, it can be seen that the mixed oil prepared by the present application has good barrier repair effect, can significantly activate the expression of FLG and LOR mRNA, and the above effects of yunnan peony seed, Chinese pricklyash fruit and camellia seed have certain mutual cooperation, and the three are indispensable, the pretreatment before fermentation can improve the technical effect of extraction, the common fermentation of aspergillus niger and bacillus pumilus can maximize the fermentation effect, and the two strains are indispensable, and the effect of subcritical extraction is better than that of other extraction effects.

[0130] Test example 4

[0131] The fatty acid content detection method refers to the national standard GB5009.168-2016, determination of fatty acids in food(normalization method).

[0132] Total flavone detection method: respectively take 0.5mL of the measured solution and rutin control solution in a colorimetric tube, add 0.2mL of 5%NaNO2, mix well, and place for 6min, then add 0.2mL of 1%Al(NO3)3, mix well, and place for 6min, then add 1mL of 4%NaOH, and place at room temperature for 15min. Measure the absorbance at 500nm wavelength, draw the standard curve of rutin standard solution, and calculate the total flavone content by substituting the absorbance of the sample into the standard curve.

[0133] The anthocyanin detection method is as follows: 0.5 mg / mL anthocyanin standard solution is prepared, 0.1, 0.2, 0.3, 0.4 and 0.5 mL of the solution are respectively taken into a colorimetric tube, and then 1.0 mL of methanol solution is added, then 6 mL of n-butanol-hydrochloric acid solution and 0.2 mL of 2% iron ammonium sulfate solution are added into the standard solution and the to-be-detected solution, and then the mixture is shaken, heated in a boiling water bath for 40 min, and then quickly cooled, the absorbance of the anthocyanin standard solution and the to-be-detected solution is measured at a wavelength of 530 nm, and then the anthocyanin content of the to-be-detected solution is calculated by drawing an anthocyanin standard curve.

[0134] Table 4

[0135] Example 1 Example 2 Example 3 Example 12 Oleic acid (%) 33.2 32.9 32.1 37.6 Linoleic acid (%) 25.6 24.2 24.4 27.6 Linolenic acid (%) 18.4 18.6 19.2 14.4 Total flavonoids (%) 0.54 0.49 0.51 0.32 Anthocyanins (%) 1.44 1.21 1.17 0.86

[0136] It can be seen from the data in Table 4 that the contents of oleic acid, linoleic acid, linolenic acid, total flavonoids and anthocyanin of the mixed oil obtained by the specific preparation method are all high, the contents of oleic acid and linoleic acid of Example 12 (without microbial fermentation) are slightly higher than those of Examples 1-3, the content of linolenic acid is lower than that of Examples 1-3, and the contents of total flavonoids and anthocyanin are lower than those of Examples 1-3, so it can be seen that microbial fermentation changes the component ratio of the mixed oil, and thus enhances the soothing, moisturizing and barrier repair effects of the mixed oil.

[0137] The applicant declares that the mixed oil for repairing sensitive skin and the preparation method and application thereof of the present application are illustrated by the above examples, but the present application is not limited to the above examples, that is, it does not mean that the present application must rely on the above examples to be implemented. It should be understood by those skilled in the art that any improvement of the present application, equivalent replacement of each raw material of the product of the present application, addition of auxiliary ingredients, selection of specific modes, etc. all fall within the protection scope and disclosure scope of the present application.

[0138] The preferred embodiments of the present application are described in detail above, but the present application is not limited to the specific details in the above embodiments, and various simple modifications can be made to the technical solutions of the present application within the technical concept of the present application, and these simple modifications all belong to the protection scope of the present application.

[0139] In addition, it should be noted that each specific technical feature described in the above specific embodiments can be combined by any suitable method without contradiction, and in order to avoid unnecessary repetition, the present application will not further describe various possible combination methods.

Claims

1. A blend of oils for repairing sensitive skin, characterized in that it comprises: The preparation raw material of the mixed oil includes 4-6 parts of Paeonia decompendula seed, 1-3 parts of Prinsepia uniflora fruit and 1-2 parts of Camellia seed by mass fraction; The mixed oil is prepared by a preparation method comprising the following steps: (1) mixing the preparation raw material with fermentation strains, fermenting, drying to obtain a fermented mixture; (2) using subcritical extraction on the fermented mixture to obtain crude oil; (3) filtering the crude oil to obtain the mixed oil; The fermentation strains comprise Aspergillus niger and Bacillus pumilus with a ratio of viable bacterial count of 1:(1-3); The extraction solvent used in the subcritical extraction is anhydrous ethanol and butane with a volume ratio of 1:(4-7).

2. The mixed oil for repairing sensitive skin according to claim 1, wherein The amount of the fermentation strain added is 2 x 10 8 CFU / g - 5 x 10 8 CFU / g.

3. The mixed oil for repairing sensitive skin according to claim 1, wherein The fermentation temperature is 30-40℃, the fermentation time is 48-96 h, and the water content of the fermented material is 30-50%.

4. The mixed oil for repairing sensitive skin according to claim 1, wherein The ratio of the fermented mixture to the extraction solvent is 1 g:(2-10) mL.

5. The mixed oil for repairing sensitive skin according to claim 1, wherein The temperature of the subcritical extraction is 35-45℃, the single extraction time is 2-3 h, the extraction times is 2-3 times, and the extraction pressure is 0.3-0.5 MPa.

6. Use of the mixed oil for repairing sensitive skin according to any one of claims 1-5 in the preparation of a product having soothing, moisturizing or barrier repair efficacy.

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