Oxygen-enriched oil compound composition, emulsion as well as preparation method and application of oxygen-enriched oil compound composition and emulsion

Abstract

The invention relates to the technical field of daily chemical products, and provides an oxygen-enriched oil compound composition, an emulsion and a preparation method and application thereof, and the composition comprises the following raw materials: oxygen-enriched oil, docosahexaenoic acid and ceramide in a mass ratio of (10-17): (0.1-1): (0.02-0.6). The oxygen-enriched oil, the docosahexaenoic acid and the ceramide cooperate with one another in function, so that the obtained composition has a synergistic effect in the aspects of antibiosis, anti-inflammation, barrier repair, oxidation resistance and the like. The components are compounded in an oil-in-oil type structure, three active components are respectively accommodated through a unique double-oil-phase structure, and the problem of coexistence stability is solved by further utilizing a nano-scale entrapment technology; and the dosage form further promotes the synergistic interaction of the oxygen-enriched oil compound composition in the aspects of antibiosis, anti-inflammation, barrier repair, oxidation resistance and the like.

Description

Technical Field

[0001] This invention relates to the technical field, and more particularly to an oxygen-enriched oil compound composition, emulsion, its preparation method, and its application. Background Technology

[0002] Docosahexaenoic acid (DHA) is an essential Omega-3 polyunsaturated fatty acid. Its chemical structure contains six double bonds, a unique polyunsaturated structure that endows DHA with distinctive physicochemical properties and biological activity. Despite its multiple skin-care benefits, the numerous unsaturated double bonds in the DHA molecule make it chemically unstable, easily oxidized to produce a rancid odor and harmful substances. Furthermore, DHA has extremely poor water solubility, making it difficult to apply directly to water-based systems and resulting in low bioavailability.

[0003] Ozone-rich oils are oil-soluble active ingredients produced by infusing ozone gas into base oils, such as olive oil. They are rich in ozonides, peroxides, and hydroxylated fatty acids. Although ozone-rich oils offer multiple skincare benefits, the ozone component is chemically unstable and prone to decomposition, leading to loss of activity and potential irritation. Furthermore, their oily matrix makes them difficult to apply directly to water-based systems, resulting in poor compatibility and potentially affecting skin feel.

[0004] Ceramides are a major component of the lipid bilayer of the stratum corneum in the skin. Their highly hydrophobic double-chain structure endows the stratum corneum with the density and selective permeability of a "brick-and-wall" barrier. Although ceramides have significant skincare benefits, their strong hydrophobicity makes them prone to crystallization in formulations, affecting product stability and efficacy. Furthermore, ceramides have poor water dispersibility, making them difficult to apply directly to conventional skincare formulations.

[0005] Studies have found that DHA is easily oxidized, oxygen-rich oil is unstable, and ceramides are prone to crystallization. Traditional formulations cannot simultaneously guarantee their stability and compatibility. No research reports on the systematic compounding of these three components have been found in existing patents and technical literature. Therefore, this invention is proposed. Summary of the Invention

[0006] To partially solve the above-mentioned technical problems, the present invention provides an oxygen-enriched oil compound composition, emulsion, preparation method and application thereof. Through the synergistic effect of oxygen-enriched oil, docosahexaenoic acid and ceramide, the resulting oxygen-enriched oil compound composition has synergistic effects in terms of immediate and long-term anti-inflammatory effects, barrier repair and improvement of skin firmness.

[0007] In a first aspect, the present invention provides an oxygen-enriched oil compound composition, the raw materials of which include: oxygen-enriched oil, docosahexaenoic acid and ceramide in a mass ratio of (10~17):(0.1~1):(0.02~0.6).

[0008] Studies have found that oxygen-enriched oil, docosahexaenoic acid (DHA), and ceramides function synergistically, exhibiting enhanced effects in antibacterial, anti-inflammatory, barrier repair, and antioxidant properties. Specifically, the potent antibacterial properties of oxygen-enriched oil create a cleaner microenvironment for DHA and ceramides, while ceramides structurally strengthen the skin barrier, providing a stable skin foundation for DHA and oxygen-enriched oil to exert their effects. Furthermore, the anti-inflammatory and immunomodulatory functions of DHA can alleviate oxidative stress that may be induced by oxygen-enriched oil, and the potential barrier damage risk from oxygen-enriched oil can be mitigated by the superior barrier repair capabilities of ceramides.

[0009] Based on this, the interaction of these three elements allows the oxygen-enriched oil compound composition of the present invention to fully utilize the technical characteristics of ozone oil and ceramides in rapidly and timely relieving inflammation, as well as the long-lasting anti-inflammatory effect of DHA, achieving rapid redness reduction and anti-inflammatory effects on the skin. Furthermore, important components of the extracellular matrix include collagen, fibronectin (FN1), and elastin (ELN). The interaction between collagen, fibronectin (FN1), and elastin (ELN) forms the supporting structure for tissue cells and is also the microenvironment for tissue cell survival, playing a crucial role in tissue cell survival, regeneration, repair, and immunity. Studies have found that the oxygen-enriched oil compound composition of the present invention can also promote the expression of collagen, fibronectin, and elastin, the main components of the skin's extracellular matrix, thereby promoting skin barrier repair, wound care, and improving skin firmness.

[0010] Preferably, the mass ratio of oxygen-enriched oil to docosahexaenoic acid is (20~60):1; for example, it can be any value or a range of values ​​from 20:1, 30:1, 40:1, 50:1, 60:1.

[0011] Preferably, the ratio of oxygen-enriched oil to ceramide (60~100):1; for example, it can be any value or a range of values ​​from 60:1, 70:1, 80:1, 90:1, 100:1.

[0012] According to the oxygen-enriched oil compound composition provided by the present invention, the oxygen-enriched oil is obtained by passing ozone into a base oil, wherein the base oil is selected from one or more of olive oil, rapeseed oil, flaxseed oil, palm oil, sunflower oil, avocado oil, *Sapindus mukorossi* oil, sesame oil, and jojoba oil.

[0013] Preferably, the oxygen-enriched oil is prepared by introducing 0.1% to 30% by weight of ozone into base oil and reacting for 20 minutes to 12 hours. The 0.1% to 30% by weight of ozone can be any value or a range of values ​​from 0.1%, 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%, 26%, 28%, and 30%.

[0014] The reaction time of 20 min to 12 h mentioned in this invention can be any value or a numerical range composed of any values ​​among 20 min, 25 min, 30 min, 1 h, 4 h, 6 h, 8 h, 10 h, and 12 h.

[0015] In the conventional case of the present invention, the ozone content in the oxygen-enriched oil to be deodorized is 0.85% to 10%.

[0016] Preferably, the oxygen-enriched oil is used in the oxygen-enriched oil compound composition after being deodorized.

[0017] Deodorization treatment helps improve the sensory properties of oxygen-enriched oils. Experiments have shown that a suitable deodorization process can improve the sensory properties of oxygen-enriched oils without reducing their efficacy.

[0018] According to the oxygen-enriched oil compound composition provided by the present invention, the deodorization treatment includes: introducing steam into the oxygen-enriched oil at a temperature of 130~150°C and deodorizing under negative pressure conditions.

[0019] Preferably, the peroxide value of the oxygen-enriched oil is greater than or equal to 100 meq / kg.

[0020] Preferably, the peroxide value of the oxygen-enriched oil after deodorization treatment is less than or equal to 10 meq / kg.

[0021] Preferably, the negative pressure condition can be selected to be below 500 Pa, maintaining a medium or higher degree of vacuum.

[0022] Preferably, the heat preservation time corresponding to deodorization is 30~60 minutes.

[0023] The oxygen-enriched oil compound composition provided by the present invention comprises, by mass percentage: 50%~85% oxygen-enriched oil, 0.5%~5% docosahexaenoic acid, and 0.1%~3% ceramide.

[0024] In this invention, the mass percentage of the oxygen-enriched oil can be any value or a range of any values ​​from 50%, 55%, 60%, 65%, 70%, 75%, 80%, to 85%.

[0025] In this invention, the mass percentage of docosahexaenoic acid can be any value or a range of any values ​​from 0.5%, 1.5%, 2.5%, 3.5%, 4.5%, to 5.0%.

[0026] In this invention, the mass percentage of the ceramide can be any value or a range of any values ​​from 0.1%, 0.5%, 1.0%, 1.5%, 2.0%, 2.5%, and 3.0%.

[0027] Preferably, by weight percentage, the oxygen-enriched oil compound composition further includes one or more of the following: antioxidants, emulsifiers, humectants, thickeners, and stabilizers.

[0028] Preferably, the antioxidant is selected from one or more of tocopherol, lecithin, rosemary extract, ascorbyl palmitate, and propyl gallate; the antioxidant is preferably a fat-soluble antioxidant.

[0029] Preferably, the emulsifier is selected from one or more of caprylic / capric triglycerides, lecithin, hydrogenated lecithin, sucrose esters, and polysorbate esters.

[0030] Preferably, the antioxidant in the oxygen-enriched oil compound composition is 0.5% to 10% by mass percentage.

[0031] Preferably, the emulsifier in the oxygen-enriched oil compound composition is 2% to 23% by mass percentage.

[0032] This invention achieves synergistic homeostasis regulation of ozonation products by compounding plant oils, bio-esters, or fat-soluble antioxidants, thereby further enhancing their antibacterial, repair, and other biological activities.

[0033] The oxygen-enriched oil compound composition of the present invention can be widely applied to fields such as skin barrier damage, inflammatory diseases, chronic wounds, antibacterial care, and functional cosmetics.

[0034] In a second aspect, the present invention also provides an emulsion comprising the oxygen-enriched oil compound composition described above.

[0035] As mentioned above, DHA is easily oxidized, oxygen-enriched oil is unstable, and ceramide is prone to crystallization. Traditional formulations cannot simultaneously guarantee their stability and compatibility. Furthermore, the properties of oxygen-enriched oil change significantly after ozone modification, making it difficult to mix with conventional oils. Research has found that the above components are combined in an oil-in-oil structure, which accommodates the three active ingredients separately through its unique two-oil phase structure. The problem of coexistence stability is further solved by utilizing nanoscale encapsulation technology. Moreover, this formulation further promotes the synergistic effect of the oxygen-enriched oil compound in antibacterial, anti-inflammatory, barrier repair, and antioxidant properties.

[0036] The experiment also found that using oxygen-enriched oil as the inner phase, combined with the other two components to form an oil-in-oil emulsion, can effectively solve the problems of strong odor, instability, and skin irritation. Simultaneously, the oil-phase nanoemulsion carrier structure avoids the degradation of ozone products by water, which helps extend shelf life and enhance skin absorption.

[0037] To ensure a more uniform mixing and a more stable structure, the emulsion preferably also includes antioxidants and / or emulsifiers, preferably lecithin.

[0038] According to the emulsion provided by the present invention, the emulsion is an oil-in-oil emulsion, the external phase of which includes docosahexaenoic acid and ceramide; and the internal phase of which includes oxygen-enriched oil.

[0039] According to the emulsion provided by the present invention, the emulsion is a nanoemulsion, and more preferably, the particle size distribution of the nanoemulsion is 20~300nm.

[0040] Thirdly, the present invention also provides a method for preparing the emulsion as described above, comprising: adding an internal phase to an external phase to obtain a crude emulsion, and homogenizing the crude emulsion to obtain the skin emulsion; wherein the external phase comprises docosahexaenoic acid and ceramide; and the internal phase comprises oxygen-enriched oil.

[0041] The method for preparing the skin emulsion according to the present invention includes: The internal phase was obtained by heating and dissolving lecithin and oxygen-enriched oil together. The external phase was obtained by heating and dissolving docosahexaenoic acid, ceramide, and emulsifier. Adding the inner phase to the outer phase yields a crude emulsion; The crude milk was homogenized to obtain the skin emulsion.

[0042] Studies have found that using lecithin to mix oxygen-enriched oil with other ingredients results in a more stable emulsion structure, which is more conducive to the better performance of the oxygen-enriched oil compound composition.

[0043] The method for preparing the skin emulsion according to the present invention includes: Lecithin and oxygen-enriched oil are mixed and heated to 40-50°C, then stirred until lecithin is completely dispersed and dissolved. The mixture is then cooled to below 30°C to obtain the internal phase. The external phase is obtained by heating docosahexaenoic acid, ceramide, and emulsifier to 40-50°C and stirring to disperse and dissolve them. The inner phase is added to the outer phase to obtain crude emulsion.

[0044] The crude emulsion was homogenized at 10,000 to 30,000 psi to obtain the skin emulsion.

[0045] The emulsion of the present invention has advantages such as high stability, strong permeability, low irritation and controllable release. It has broad application prospects in the fields of skin barrier repair, wound care, acne treatment and other skin health management. It can also be used as a functional active oil raw material in cosmetics, medical dressings and so on, and has significant industrialization value and promotion prospects.

[0046] Fourthly, the present invention also provides the use of the oxygen-enriched oil compound composition as described above, the emulsion as described above, and the emulsion prepared by the method of preparing the emulsion as described above in cosmetics.

[0047] Preferably, the dosage form of the cosmetic is selected from serum, emulsion or cream.

[0048] Preferably, the oxygen-enriched oil compound composition is used for one or more of the following purposes: promoting the expression of collagen in the skin extracellular matrix, promoting the expression of fibronectin in the skin extracellular matrix, promoting the expression of elastin in the skin extracellular matrix, and promoting anti-inflammatory repair of the skin.

[0049] The oxygen-enriched oil compound composition, emulsion, preparation method and application provided by the present invention, through the synergistic effect of oxygen-enriched oil, docosahexaenoic acid and ceramide, enable the obtained oxygen-enriched oil compound composition to have synergistic effects in antibacterial and anti-inflammatory, barrier repair and antioxidant properties.

[0050] The above-mentioned components are combined in an oil-in-oil structure, which solves the problem of coexistence stability. Moreover, this formulation further promotes the synergistic effect of the oxygen-rich oil compound in antibacterial, anti-inflammatory, barrier repair and antioxidant properties. Detailed Implementation

[0051] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of this invention, not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0052] Where specific techniques or conditions are not specified in the embodiments, they shall be performed in accordance with the techniques or conditions described in the literature in this field, or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased through legitimate channels. The models or parameters of some of the raw materials involved in the embodiments of this invention are shown below: Oxygen-enriched oil: POV value of 424 meq / k, ozone content of 1.5%.

[0053] DHA lipids: The DHA content is 45%, and the remaining components are mainly medium and long chain saturated fatty acids.

[0054] Example 1 This embodiment provides a skin-use nanoemulsion with an oil-in-oil (O / O) structure. The inner phase consists of oxygen-rich oil and lecithin, while the outer phase comprises DHA oil, ceramides, tocopherol, and caprylic / capric triglycerides. The mass percentages of each component in the skin-use nanoemulsion are as follows: It contains 80% oxygen-enriched oil, 2% DHA oil, 1% ceramide, 2% tocopherol, 5% lecithin, and 10% caprylic / capric triglycerides.

[0055] This embodiment also provides a method for preparing the above-mentioned skin nanoemulsion, the steps of which are as follows: (1) Mix lecithin and oxygen-enriched oil, heat to 40°C and stir until lecithin is completely dispersed and dissolved, then cool to below 30°C to obtain the internal phase.

[0056] (2) The external phase is obtained by heating DHA oil, ceramide, tocopherol, caprylic / capric triglyceride to 40°C and stirring to disperse and dissolve.

[0057] (3) Slowly add the internal phase to the external phase and stir until homogeneous to obtain crude emulsion.

[0058] (4) The crude emulsion was homogenized under high pressure using a high-pressure microfluidic homogenizer (ATS) for 3 cycles at 20,000 psi to finally obtain an oil-in-oil (O / O) structured skin nanoemulsion with a particle size distribution of 20~300 nm.

[0059] Example 2 This embodiment provides a skin-use nanoemulsion with an oil-in-oil (O / O) structure. The inner phase consists of oxygen-rich oil and lecithin, while the outer phase comprises DHA, ceramides, tocopherol, and caprylic / capric triglycerides. The mass percentages of each component in the skin-use nanoemulsion are as follows: The deodorized oxygen-enriched oil contains 80%, DHA oil 2%, ceramide 1%, tocopherol 2%, lecithin 5%, and caprylic / capric triglycerides 10%.

[0060] This embodiment also provides a method for preparing the above-mentioned skin nanoemulsion, the steps of which are as follows: (1) The lecithin and deodorized oxygen-enriched oil were mixed and heated to 40°C and stirred until the lecithin was completely dispersed and dissolved. The mixture was then cooled to below 30°C to obtain the internal phase. The deodorized oxygen-enriched oil was obtained by passing pure steam into the oxygen-enriched oil at 130°C and deodorizing it under a vacuum of 300Pa. The holding time for deodorization was 60 min, and the POV value was measured to be 4.3 meq / kg.

[0061] (2) The external phase is obtained by heating DHA oil, ceramide, tocopherol, caprylic / capric triglyceride to 40°C and stirring to disperse and dissolve.

[0062] (3) Slowly add the internal phase to the external phase and stir until homogeneous to obtain crude emulsion.

[0063] (4) The crude emulsion was homogenized under high pressure using a high-pressure microfluidic homogenizer (ATS) for 3 cycles at 20,000 psi to finally obtain an oil-in-oil (O / O) structured skin nanoemulsion with a particle size distribution of 20~300 nm.

[0064] Example 3 This embodiment provides a skin-use nanoemulsion with an oil-in-oil (O / O) structure. The inner phase consists of oxygen-rich oil and lecithin, while the outer phase comprises DHA, ceramides, tocopherol, and caprylic / capric triglycerides. The mass percentages of each component in the skin-use nanoemulsion are as follows: The deodorized oxygen-enriched oil contains 80%, DHA oil 2%, ceramide 1%, tocopherol 2%, lecithin 5%, and caprylic / capric triglycerides 10%.

[0065] This embodiment also provides a method for preparing the above-mentioned skin nanoemulsion, the steps of which are as follows: (1) The lecithin and deodorized oxygen-enriched oil were mixed and heated to 40-50°C and stirred until the lecithin was completely dispersed and dissolved. The mixture was then cooled to below 30°C to obtain the internal phase. The deodorized oxygen-enriched oil was obtained by passing pure steam into the oxygen-enriched oil at 140°C and deodorizing it under a vacuum of 250 Pa. The holding time for deodorization was 30 min, and the POV value was measured to be 6.3 meq / k.

[0066] (2) The external phase is obtained by heating DHA oil, ceramide, tocopherol, caprylic / capric triglyceride to 45°C and stirring to disperse and dissolve.

[0067] (3) Slowly add the internal phase to the external phase and stir until homogeneous to obtain crude emulsion.

[0068] (4) The crude emulsion was homogenized under high pressure using a high-pressure microfluidic homogenizer (ATS) for 3 cycles at 20,000 psi to finally obtain an oil-in-oil (O / O) structured skin nanoemulsion with a particle size distribution of 20~300 nm.

[0069] Comparative Example 1 This comparative example provides a skin-use nanoemulsion, whose raw materials, by mass percentage, are as follows: It contains 80% oxygen-enriched oil, 3% ceramide, 2% tocopherol, 5% lecithin, and 10% caprylic / capric triglycerides.

[0070] This comparative example also provides a method for preparing the above-mentioned skin nanoemulsion, the preparation steps of which are as follows: (1) Mix lecithin and oxygen-enriched oil, heat to 45°C and stir until lecithin is completely dispersed and dissolved, then cool to below 30°C to obtain the internal phase.

[0071] (2) Ceramide, tocopherol, and caprylic / capric triglyceride are heated to 40-50°C and stirred to disperse and dissolve to obtain the external phase.

[0072] (3) Slowly add the internal phase to the external phase and stir until homogeneous to obtain crude emulsion.

[0073] (4) The crude emulsion was homogenized under high pressure using an ATS (Advanced Pressure Surface Heater) for 3 cycles at 20,000 psi to finally obtain an oil-in-oil (O / O) nanoemulsion for skin.

[0074] Comparative Example 2 This comparative example provides a skin emulsion, the raw materials of which are as follows by weight percentage: DHA lipids 83%, tocopherol 2%, lecithin 5%, caprylic / capric triglycerides 10%.

[0075] This comparative example also provides a method for preparing the above-mentioned skin nanoemulsion, the preparation steps of which are as follows: (1) DHA oil, lecithin, tocopherol, and caprylic / capric triglycerides are heated to 40-50°C and stirred to disperse and dissolve to obtain crude emulsion.

[0076] (2) The crude emulsion was homogenized under high pressure using an ATS (Advanced Pressure Streamer) at 20,000 psi for 3 cycles to finally obtain DHA emulsion.

[0077] Comparative Example 3 A skin-use blended emulsion, comprising the following ingredients by weight percentage: Tocopherol 2%, lecithin 5%, and caprylic / capric triglycerides 93%.

[0078] This comparative example also provides a method for preparing the above-mentioned skin-use mixed emulsion, the preparation steps of which are as follows: (1) Mix lecithin, tocopherol and caprylic / capric triglycerides at 40-50°C to obtain crude milk.

[0079] (2) The crude emulsion was homogenized under high pressure using an ATS (Advanced Pressure Surface Heater) for 3 cycles at 20,000 psi to obtain a mixed solution for skin use.

[0080] Test Example 1: Skin Anti-allergy Verification This experiment aims to induce predictable, transient stinging, burning, and visible erythema using a human lactic acid stinging model by applying a high concentration of lactic acid (10%) to the skin. By comparing the speed and extent of the resolution of these symptoms after application, its immediate anti-inflammatory and soothing effects can be directly demonstrated. The specific experimental arrangements are as follows: Subjects: Forty healthy volunteers who were pre-screened as sensitive (i.e., had a clear stinging sensation) by lactic acid stinging test were randomly divided into four groups of 10 each.

[0081] Test area: The same test area was drawn on both cheeks of each subject.

[0082] Experimental Groups: Group A: Apply the mixed solution to the skin as described in Comparative Example 3.

[0083] Group B: Apply the skin nanoemulsion of Example 1.

[0084] Group C: Apply nanoemulsion to the skin of Comparative Example 2.

[0085] Group D: Apply nanoemulsion to the skin of Comparative Example 1.

[0086] The experimental procedure is as follows: (1) Baseline value measurement: The initial a* value (erythema index) of all test areas was measured using a chromameter, and the subject’s baseline discomfort (0 points for no discomfort, 10 points for severe stinging) was recorded.

[0087] (2) Induction stimulation: Apply 10% lactic acid solution evenly to all test areas, leave for 5 minutes, and then gently wash off with water.

[0088] (3) Initial assessment and product application: Immediately after stimulation, the erythema index (a*) and the subject's subjective stinging score (T0) were measured. Immediately after measurement, a sufficient amount (2 mg / cm²) was applied to the corresponding area of ​​each group. 2 The test substance.

[0089] (4) Effect evaluation: 5 minutes (T5) and 15 minutes (T6) after application of the test substance.15 ) and 30 minutes (T 30 Measure the a* value again and record the subjective score.

[0090] The test results for the a* value are shown in Table 1 below.

[0091] Table 1

[0092] Note: Compared with Group A at the same time point, *P<0.05, **P<0.01, ***P<0.001.

[0093] The test results of subjective scoring are shown in Table 2 below.

[0094] Table 2

[0095] Note: Compared with Group A at the same time point, *P<0.05, **P<0.01, ***P<0.001.

[0096] The results of this experiment show that Group B (the skin nanoemulsion of Example 1) can rapidly and significantly reduce erythema and subjective discomfort in a lactic acid-induced acute skin irritation model, and its immediate anti-inflammatory and soothing effect far exceeds that of any single or partial combination of ingredients.

[0097] Test Example 2: Anti-inflammatory Experiment Monocytes and macrophages (THP-1) were stimulated with 1 μg / mL lipopolysaccharide (LPS). The soothing efficacy of the test samples was evaluated by detecting changes in the levels of inflammatory factors (IL-1β, IL-6, and PGE2) after sample treatment. The specific procedure is as follows: (1) Experimental materials: THP-1 cells, batch number: STM-CL-5011.

[0098] 1640 culture medium, fetal bovine serum, PBS, dexamethasone, IL-1β ELISA kit, IL-6 (interleukin-6) ELISA kit, PGE2 ELISA kit, lipopolysaccharide, MTT, DMSO, phorbol.

[0099] CO2 incubator, clean bench, microplate reader (BioTek, Epoch), inverted microscope (Olympus, CKX41).

[0100] (2) Working fluid The 1 μg / mL LPS diluent is prepared by diluting a 2 mg / mL LPS stock solution.

[0101] 0.01% dexamethasone is prepared by dissolving 2 μL of 10% stock solution in 2 mL of culture medium (1640 medium).

[0102] (3) Experimental grouping: Group BC: Blank control group.

[0103] PC group: 0.01% dexamethasone.

[0104] NC group: Comparative example 3: skin nanoemulsion.

[0105] DHA group: Comparative example 2: skin nanoemulsion.

[0106] OC group: Comparative Example 1: Skin nanoemulsion.

[0107] OCD Group: Skin Nanoemulsion of Example 1.

[0108] dOCD group: Skin nanoemulsion of Example 2.

[0109] (4) Experimental methods (4.1) Cell seeding: Cells were seeded in 12-well plates at 2 mL / well and 5E5 cells / mL. The cells were suspended in fresh medium containing 150 ng / mL PMA and activated for 72 h.

[0110] (4.2) Administration: Design the following experiment: Table 3

[0111] Drug administration process: The 12-well plate was washed three times with PBS, and fresh culture medium was added. The test samples were added according to the experimental groups and their corresponding concentrations. Except for the BC group, which did not have LPS induction, all other groups were LPS induced.

[0112] (3.3) After administration, the 12-well plate was placed in a cell culture incubator (5% CO2, 37℃) for 24 hours.

[0113] (4) Test After incubation for 24 hours following drug administration, the cell culture supernatant was collected. After collection, the samples for ELISA detection were frozen at -80°C and analyzed according to the instructions of the IL-1β ELISA kit, IL-6 ELISA kit, and PGE2 ELISA kit.

[0114] (5) Results Analysis A standard curve was plotted with the concentration of the standard as the x-axis and the OD value as the y-axis, and the regression equation was calculated. The concentration (pg / mL) of each inflammatory factor in each sample group was calculated by substituting the OD value of each sample group into the equation.

[0115] Data processing: Statistical analysis was performed using software. Results are expressed as mean ± SD. One-way ANOVA or t-test was used for comparisons between groups.

[0116] The test results for IL-1β, IL-6, and PGE2 levels are shown in the table below.

[0117] Table 4 IL-1β content

[0118] Table 5 IL-6 content

[0119] Table 6 PGE2 content

[0120] In Tables 3-5 above, when performing statistical analysis using the t-test method, the significance of comparison with the BC group is indicated by #, with P-value < 0.05 indicated by # and P-value < 0.01 indicated by ##; the significance of comparison with the NC group is indicated by *, with P-value < 0.05 indicated by * and P-value < 0.01 indicated by **.

[0121] Test Example 3: Detection of Extracellular Matrix Components Using fibroblasts as the testing tool, this study evaluates the promoting effect of the test sample on extracellular matrix components and its effect on skin firming and anti-wrinkle by detecting changes in the content of collagen, fibroin, and elastin after the sample is applied to fibroblasts.

[0122] The testing method is as follows: (1) Experimental materials: Fibroblasts (STM-CL-5176 Stemer).

[0123] TGFβ (transforming growth factor β), DMEM culture medium, fetal bovine serum, PBS, DMSO, Collagen I ELISA kit, elastin (ELN) ELISA kit and FN1 ELISA kit.

[0124] CO2 incubator, clean bench, microplate reader (BioTek, Epoch), inverted microscope (Olympus, CKX41).

[0125] Working fluid: Group BC: Comparative Example 3: Skin nanoemulsion.

[0126] PC group: TGFβ.

[0127] OC group: Comparative Example 1: Skin nanoemulsion.

[0128] DHA group: Comparative example 2: skin nanoemulsion.

[0129] OCD Group: Skin Nanoemulsion of Example 1.

[0130] dOCD group: Skin nanoemulsion of Example 2.

[0131] (2) Experimental design: (2.1) Cell seeding: at 1.0 × 10 5 Fibroblasts were seeded at a density of cells / well into 96-well plates and incubated overnight in an incubator (37°C, 5% CO2). (2.2) Drug administration: The following experiment was designed: Table 7

[0132] Drug administration process: The sample was dissolved in DMSO at a 1:1 ratio to prepare a 5% stock solution. When the cell layering rate in the 96-well plate reached 40%~50%, the drug was administered according to the sample concentration. Each group was divided into 3 replicates and cultured in an incubator (37℃, 5% CO2) for 24 hours.

[0133] (2.3) Collect cell supernatant: After culturing for 24 hours, collect the cell culture supernatant in an EP tube. After collection, place the sample for Collagen I content detection in a -80℃ freezer for storage.

[0134] (2.4) Detection of collagen, fibroin and elastin content: The detection was carried out according to the instructions of the Collagen I ELISA kit, the elastin (ELN) ELISA kit and the FN1 ELISA kit.

[0135] The test results of collagen, fibroin and elastin content are shown in the table below. The t-test was used for statistical analysis between the groups. Compared with the blank group, P < 0.05 indicates a significant difference (*), and P < 0.01 indicates an extremely significant difference (**).

[0136] Table 8

[0137] Table 9

[0138] Table 10

[0139] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. An oxygen-enriched oil compound composition, characterized in that, Its raw materials include: oxygen-enriched oil, docosahexaenoic acid and ceramide in a mass ratio of (10~17):(0.1~1):(0.02~0.6).

2. The oxygen-enriched oil compound composition according to claim 1, characterized in that, The oxygen-enriched oil is obtained by passing ozone into a base oil, wherein the base oil is selected from one or more of olive oil, rapeseed oil, flaxseed oil, palm oil, sunflower oil, avocado oil, sage fruit oil, sesame oil, and jojoba oil. Preferably, the oxygen-enriched oil is used in the oxygen-enriched oil compound composition after being deodorized.

3. The oxygen-enriched oil compound composition according to claim 2, characterized in that, The deodorization process includes: introducing steam into oxygen-enriched oil at a temperature of 130~150℃ and deodorizing under negative pressure conditions; Preferably, the peroxide value of the oxygen-enriched oil is greater than or equal to 100 meq / kg; Preferably, the peroxide value of the oxygen-enriched oil after deodorization treatment is less than or equal to 10 meq / kg.

4. The oxygen-enriched oil compound composition according to any one of claims 1 to 3, characterized in that, By weight percentage, its raw materials include: 50%~85% oxygen-enriched oil, 0.5%~5% docosahexaenoic acid, and 0.1%~3% ceramide.

5. An emulsion, characterized in that, Includes the oxygen-enriched oil compound composition according to any one of claims 1 to 4.

6. The emulsion according to claim 5, characterized in that, The emulsion is an oil-in-oil emulsion, the external phase of which includes docosahexaenoic acid and ceramide; and the internal phase of which includes oxygen-enriched oil.

7. The emulsion according to claim 5 or 6, characterized in that, The emulsion is a nanoemulsion, and more preferably, the particle size distribution of the nanoemulsion is 20~300nm.

8. A method for preparing the emulsion according to any one of claims 5 to 7, characterized in that, include: The inner phase is added to the outer phase to obtain a crude emulsion, and the crude emulsion is homogenized to obtain the skin emulsion; The external phase comprises docosahexaenoic acid and ceramide; the internal phase comprises oxygen-enriched oil.

9. The method for preparing the skin emulsion according to claim 8, characterized in that, include: The internal phase was obtained by heating and dissolving lecithin and oxygen-enriched oil together. The external phase was obtained by heating and dissolving docosahexaenoic acid, ceramide, and emulsifier. Adding the inner phase to the outer phase yields a crude emulsion; The crude milk was homogenized to obtain the skin emulsion.

10. Use in cosmetics of the oxygen-enriched oil compound composition according to any one of claims 1 to 4, the emulsion according to any one of claims 5 to 7, and the emulsion prepared by the method of claim 8 or 9; Preferably, the oxygen-enriched oil compound composition is used for one or more of the following purposes: promoting the expression of collagen in the skin extracellular matrix, promoting the expression of fibronectin in the skin extracellular matrix, promoting the expression of elastin in the skin extracellular matrix, and promoting anti-inflammatory repair of the skin.

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