Long-term stable transdermal vitamin C liposome as well as preparation method and preparation application thereof

Abstract

The invention discloses a long-term stable transdermal vitamin C liposome as well as a preparation method and preparation application thereof, belongs to the technical field of liposome, and solves the problem of single application range of vitamin C in the prior art. The liposome comprises the following components in parts by mass: 1-20 parts of an emulsifier, 1-10 parts of lipid, 1-10 parts of vitamin C, 1-4 parts of a compound functional substance and the balance of water, wherein the total mass of the components is 100 parts; wherein the compound functional substance is any one or a combination of more of ginsenoside Rg3, ginsenoside Rh2 and gentiana macrophylla A. The vitamin C liposome provided by the invention can synergistically enhance the oxidation resistance, and can further expand the application prospect of vitamin C in the daily chemical industry.

Description

Technical Field

[0001] The present invention relates to the technical field of liposomes, and more specifically to a long-term stable transdermal vitamin C liposome and a preparation method and preparation application thereof. Background Art

[0002] Vitamin C (VC), also known as L-ascorbic acid, is an important and special water-soluble vitamin, often used as a food additive and the main ingredient of cosmetics. It can be used as a vitamin supplement and an antioxidant in cosmetics; VC can prevent skin damage caused by ultraviolet rays, thereby inhibiting wrinkles, and can inhibit the formation of histamine to reduce allergic reactions, while also inhibiting the formation of skin melanin to prevent and treat skin aging. However, vitamin C is particularly sensitive to external environmental factors such as oxygen, heat and light, and is easily oxidized and decomposed. The stability of vitamin C can be enhanced through liposome encapsulation technology. At the same time, due to the strong biocompatibility of liposomes, the absorption rate of vitamin C can be enhanced, which can greatly improve the application prospects of vitamin C in daily cosmetics.

[0003] Patent CN101474156A prepares vitamin C liposomes through liposome embedding technology, and its stability is higher than that of un-embedded vitamin C, which proves that liposome embedding technology can enhance the stability of vitamin C. However, when it is compounded with other functional substances, carcinogenic organic substances such as chloroform are used, which is not conducive to industrial production and human use. Patent CN106256345A discloses a vitamin C liposome, and experiments have shown that embedding vitamin C liposomes can improve the stability and bioavailability of vitamin C, but this method is limited to using it as an oral liquid, and is not applicable when using some inedible active ingredients.

[0004] In the prior art, the application scope of vitamin C is relatively single, and multiple active ingredients are not used synergistically. Summary of the invention

[0005] The invention provides a long-term stable transdermal vitamin C liposome and a preparation method and preparation application thereof, so as to solve the problem of single application range of vitamin C in the prior art.

[0006] In the first aspect, the present invention provides a vitamin C liposome, which comprises the following components based on 100 parts by total mass: 1 to 20 parts by emulsifier, 1 to 10 parts by lipid, 1 to 10 parts by vitamin C, 1 to 4 parts by compound functional ingredient, and the rest by water; wherein the compound functional ingredient is any one or a combination of ginsenoside Rg3, ginsenoside Rh2, and gentiana macrophylla.

[0007] As a possible implementation method, the following components are included: 5 to 15 parts of emulsifier, 1 to 8 parts of lipid, 2 to 8 parts of vitamin C, 1 to 4 parts of compound functional ingredients, and the rest is water.

[0008] As a possible implementation method, the following components are included: 5 parts of emulsifier, 4 parts of lipid, 4 parts of vitamin C, 3 parts of compound functional ingredients, and the rest is water.

[0009] As a possible implementation method, the emulsifier is any one or a combination of polyglyceryl-10 stearate, PEG-8 caprylic / capric glyceride, sucrose stearate, cetearyl glucoside, stearyl alcohol PEG-2, and polysorbate-80; the lipid is any one or a combination of cholesterol, soy lecithin, hydrogenated lecithin, egg yolk lecithin, phosphatidylcholine, and phosphatidylserine.

[0010] In a second aspect, the present invention provides a method for preparing the vitamin C liposomes described in any possible implementation of the first aspect, comprising the following steps: dissolving the lipid in ethanol to obtain an ethanol solution of the lipid, evaporating the ethanol solution of the lipid under reduced pressure to form a thin film on the inner wall of a container; repeating the previous step 1 to 3 times in the same container to obtain a container with a lipid film attached to the inner wall; uniformly dissolving the emulsifier, the vitamin C and the compound functional substance in water at 30 to 50° C. to obtain a mixed solution; adding the mixed solution to the container with a lipid film attached to the inner wall, stirring to obtain a uniform mixed solution; shearing the uniform mixed solution at room temperature to obtain liposome colostrum; homogenizing the liposome colostrum several times under a high-pressure homogenizer, cooling to room temperature, to obtain the vitamin C liposomes.

[0011] As a possible implementation method, the uniform dissolution in water is carried out under the following conditions: stirring at a rotation speed of 100 to 500 rpm for 30 to 60 min; and / or, the stirring to obtain a uniform mixed liquid is carried out under the following conditions: stirring at a rotation speed of 100 to 500 rpm for 30 to 60 min; and / or, the shearing is carried out under the following conditions: shearing at a rotation speed of 8000 to 12000 rpm for 2 to 5 min; and / or, the homogenization is carried out under the following conditions: pressure of 500 to 1200 bar and temperature of 40 to 60°C.

[0012] In a third aspect, the present invention provides a use of the vitamin C liposome described in any possible implementation of the first aspect or the vitamin C liposome prepared by the preparation method described in any possible implementation of the second aspect in preparing an antioxidant preparation.

[0013] As a possible implementation, the antioxidant preparation is prepared into any one of a solution, a lotion, an ointment, a powder, a pill, a tablet, a film, an ointment, a gel, a suppository, a paste, an aerosol, and a spray.

[0014] In a fourth aspect, the present invention provides an antioxidant preparation comprising the following components: the vitamin C liposomes described in any possible implementation of the first aspect or the vitamin C liposomes prepared by the preparation method described in any possible implementation of the second aspect and excipients.

[0015] As a possible implementation method, the raw materials are measured by mass, and the preparation method includes the following steps: 20 parts of the vitamin C liposomes, 10 parts of 1% sodium hyaluronate, 10 parts of glycerol, and 20 parts of water are stirred at 50°C and 500rpm for 30 minutes to form a uniform mixed solution a; 10 parts of water, 10 parts of glycerol, 2 parts of soybean lecithin, and 5 parts of hydrogenated lecithin are mixed, and sheared under a shearing machine at a shear speed of 10000rpm and a shear time of 3min to form a uniform mixed emulsion b; the mixed solution a and the mixed emulsion b are stirred at 37°C and 500rpm for 30min to be evenly mixed to obtain the antioxidant preparation.

[0016] In order to further expand the application scenarios of vitamin C and give full play to its excellent whitening and antioxidant properties, the present invention enhances the antioxidant performance of vitamin C and slows down the self-oxidation rate of vitamin C through a compound combination of vitamin C and oil-soluble anti-inflammatory and antioxidant active ingredients ginsenoside Rg3, ginsenoside Rh2, and gentiana macrophylla, thereby enhancing the bioavailability. Finally, a liposome composition with anti-inflammatory, soothing and whitening properties is prepared through liposome encapsulation technology; while improving the compounding stability of vitamin C in various cosmetic and skin care product systems, multiple active ingredients synergize to enhance its functional properties, thereby further expanding the application prospects of vitamin C in the daily chemical industry.

[0017] In the preparation method of vitamin C liposomes provided by the present invention, the lipid ethanol solution is repeatedly evaporated under reduced pressure for multiple times, so that the lipid content and the embedding rate of active substances can be increased; anti-inflammatory and antioxidant functional substances are added for compounding to protect the antioxidant effect of vitamin C; high-speed shearing is used to simply embed the functional components, so that the efficiency of oxidative decomposition of the functional components before being homogenized into stable liposomes can be reduced; and by combining high-speed shearing with high-pressure homogenization, the uniformity of the liposomes can be effectively guaranteed, and the particle size of the liposomes can be regulated. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0019] Figure 1 The standard curve of the ABTS free radical scavenging rate of vitamin C provided in the embodiment of the present invention.

[0020] Figure 2 These are the experimental results of ABTS free radical scavenging rate of liposomes I to VIII provided in the embodiments of the present invention.

[0021] Figure 3 The results of the HaCaT cell antioxidant experiment of liposomes I to VIII provided in the examples of the present invention were compared with the H2O2 model group, *P<0.05, **P<0.01, ***P<0.005, ****P<0.0001.

[0022] Figure 4 These are the anti-inflammatory experimental results of RAW264.7 cells of liposomes I to VIII provided in the examples of the present invention.

[0023] Figure 5 These are the experimental results of tyrosinase activity detection of liposomes I to VIII provided in the embodiments of the present invention.

[0024] Figure 6 The application experiment results of liposome IV provided in the embodiments of the present invention. DETAILED DESCRIPTION

[0025] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. 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.

[0026] In order to solve the problem of single application scope of vitamin C in the prior art, the embodiment of the present invention provides a long-term stable transdermal vitamin C liposome and a preparation method and preparation application thereof.

[0027] The technical solution of the present invention will be further described below in conjunction with specific embodiments.

[0028] Example 1

[0029] This example provides a liposome preparation experiment.

[0030] 2 parts of cholesterol were added to the ethanol solution, and magnetically stirred at a speed of 300 rpm and 50°C to form a uniform lipid ethanol solution, and then the lipid ethanol solution was evaporated under reduced pressure to form a beaker Ⅰ with a lipid film on the inner wall; 3 parts of polyglycerol-10 stearate, 5 parts of VC, and 1 part of ginsenoside Rg3 were added to water for dissolution, and magnetically stirred at 300 rpm for 30 minutes at 40°C to form a uniform solution Ⅰ; the uniform solution Ⅰ was added to beaker Ⅰ, and magnetically stirred at 40°C and 200 rpm for 40 minutes to form a uniform mixed solution Ⅰ; the uniform mixed solution Ⅰ was high-speed sheared at room temperature for 5 minutes at a shear rate of 8000 rpm to obtain liposome colostrum Ⅰ; the liposome colostrum Ⅰ was homogenized twice under a high-pressure homogenizer at a homogenization pressure of 500 bar and a temperature of 40°C, and cooled to room temperature to obtain liposome Ⅰ.

[0031] 2 parts of soybean lecithin were added to the ethanol solution, and magnetically stirred at a speed of 300 rpm and 50°C to form a uniform lipid ethanol solution, and then the lipid ethanol solution was evaporated under reduced pressure; the above steps were repeated twice to form a beaker II with a lipid film on the inner wall; 2 parts of polyglycerol-10 stearate, 2 parts of sucrose stearate, 4 parts of VC, and 2 parts of ginsenoside Rg3 were added to water for dissolution, and magnetically stirred at 300 rpm for 30 minutes at 40°C to form a uniform solution II; the uniform solution II was added to beaker II, and magnetically stirred at 40°C and 200 rpm for 40 minutes to form a uniform mixed solution II; the uniform mixed solution II was high-speed sheared at room temperature for 10 minutes, with a shear rate of 9000 rpm, to obtain liposome colostrum II; the liposome colostrum II was homogenized 3 times under a high-pressure homogenizer, with a homogenization pressure of 700 bar and a temperature of 45°C, and cooled to room temperature to obtain liposome II.

[0032] 3 parts of hydrogenated lecithin were added to the ethanol solution, and magnetically stirred at a speed of 300 rpm and 45°C to form a uniform lipid ethanol solution, and then the lipid ethanol solution was evaporated under reduced pressure; the above steps were repeated twice to form a beaker III with a lipid film on the inner wall; 2 parts of polyglycerol-10 stearate, 1 part of sucrose stearate, 1 part of stearyl alcohol PEG-2, 5 parts of VC, and 2 parts of ginsenoside Rh2 were added to water for dissolution, and magnetically stirred at 300 rpm for 30 minutes at 40°C to form a uniform solution III; the uniform solution III was added to beaker III, and magnetically stirred at 50°C and 300 rpm for 30 minutes to form a uniform mixed solution III; the uniform mixed solution III was high-speed sheared at room temperature for 5 minutes, with a shear rate of 10000 rpm, to obtain liposome colostrum III; the liposome colostrum III was homogenized 4 times under a high-pressure homogenizer, with a homogenization pressure of 900 bar and a temperature of 45°C, and cooled to room temperature to obtain liposome III.

[0033] 3 parts of cholesterol were added to the ethanol solution, and magnetic stirring was performed at a speed of 300 rpm and 45°C to form a uniform lipid ethanol solution, and then the lipid ethanol solution was evaporated under reduced pressure; the above steps were repeated 3 times to form a beaker IV with a lipid film on the inner wall; 2 parts of polyglycerol-10 stearate, 1 part of sucrose stearate, 1 part of stearyl alcohol PEG-2, 1 part of cetearyl glucoside, 5 parts of VC, 1 part of ginsenoside Rg3, and 1 part of gentianin were added to water and dissolved, and stirred at 45°C. A uniform solution IV is formed by magnetic stirring at 300 rpm for 60 minutes; the uniform solution IV is added into beaker IV, and magnetically stirred at 50°C and 300 rpm for 30 minutes to form a uniform mixed solution IV; the uniform mixed solution IV is high-speed sheared at room temperature for 10 minutes at a shear rate of 12000 rpm to obtain liposome colostrum IV; the liposome colostrum IV is homogenized 6 times under a high-pressure homogenizer at a homogenization pressure of 1200 bar and a temperature of 45°C, and then cooled to room temperature to obtain liposome IV.

[0034] 3 parts of cholesterol were added to the ethanol solution, and magnetic stirring was performed at a speed of 300 rpm and 45°C to form a uniform lipid ethanol solution, and then the lipid ethanol solution was evaporated under reduced pressure; the above steps were repeated 3 times to form a beaker IV-a with a lipid film on the inner wall; 2 parts of polyglycerol-10 stearate, 1 part of sucrose stearate, 1 part of stearyl alcohol PEG-2, 1 part of cetearyl glucoside, 1 part of ginsenoside Rg3, and 1 part of gentiana were added to water and dissolved, and the mixture was stirred at 45°C by 300 rpm. The uniform solution IV-a was formed by magnetic stirring for 60 minutes; the uniform solution IV-a was added into beaker IV-a, and magnetic stirring was carried out at 50°C and 300 rpm for 30 minutes to form a uniform mixed solution IV-a; the uniform mixed solution IV-a was subjected to high-speed shearing at room temperature for 10 minutes, and the shear rate was 12000 rpm to obtain liposome colostrum IV-a; the liposome colostrum IV-a was homogenized 6 times under a high-pressure homogenizer, and the homogenization pressure was 1200 bar and the temperature was 45°C, and then cooled to room temperature to obtain liposome IV-a.

[0035] 2 parts of egg yolk lecithin were added to an ethanol solution, and magnetically stirred at a speed of 300 rpm and 50°C to form a uniform lipid ethanol solution, and then the lipid ethanol solution was evaporated under reduced pressure to form a beaker V with a lipid film on the inner wall; 3 parts of polyglycerol-10 stearate and 5 parts of VC were added to water for dissolution, and magnetically stirred at 300 rpm for 30 minutes at 40°C to form a uniform solution V; the uniform solution V was added to beaker V, and magnetically stirred at 40°C and 200 rpm for 40 minutes to form a uniform mixed solution V; the mixed solution V was high-speed sheared at room temperature for 5 minutes at a shear rate of 8000 rpm to obtain liposome colostrum V; the liposome colostrum V was homogenized once under a high-pressure homogenizer at a homogenization pressure of 300 bar and a temperature of 40°C, and cooled to room temperature to obtain liposome V.

[0036] 2 parts of phosphatidylcholine were added to the ethanol solution, and magnetically stirred at a speed of 300 rpm and 50°C to form a uniform lipid ethanol solution, and then the lipid ethanol solution was evaporated under reduced pressure; the above steps were repeated twice to form a beaker VI with a lipid film on the inner wall; 2 parts of polyglycerol-10 stearate, 2 parts of sucrose stearate, 5 parts of VC, and 1 part of ginsenoside Rg3 were added to water for dissolution, and magnetically stirred at 300 rpm for 30 minutes at 40°C to form a uniform solution VI; the uniform solution VI was added to beaker VI, and magnetically stirred at 40°C and 200 rpm for 40 minutes to form a uniform mixed solution VI; the mixed solution VI was high-speed sheared at room temperature for 10 minutes at a shear rate of 10000 rpm to obtain liposome colostrum VI; the liposome colostrum VI was homogenized twice under a high-pressure homogenizer at a homogenization pressure of 800 bar and a temperature of 45°C, and cooled to room temperature to obtain liposome VI.

[0037] 3 parts of phosphatidylserine were added to the ethanol solution, and magnetically stirred at a speed of 300 rpm and 45°C to form a uniform lipid ethanol solution, and then the lipid ethanol solution was evaporated under reduced pressure; the above steps were repeated twice to form a beaker VII with a lipid film on the inner wall; 2 parts of sucrose stearate, 2 parts of stearyl alcohol PEG-2, 5 parts of VC, and 1 part of ginsenoside Rg3 were added to water for dissolution, and magnetically stirred at 300 rpm for 30 minutes at 40°C to form a uniform solution VII; the uniform solution VII was added to beaker VII, and magnetically stirred at 50°C and 300 rpm for 30 minutes to form a uniform mixed solution VII; the mixed solution VII was high-speed sheared at room temperature for 15 minutes at a shear rate of 10000 rpm to obtain liposome colostrum VII; the liposome colostrum VII was homogenized twice under a high-pressure homogenizer at a homogenization pressure of 1000 bar and a temperature of 45°C, and cooled to room temperature to obtain liposome VII.

[0038] 3 parts of cholesterol were added to the ethanol solution, and magnetic stirring was performed at a speed of 300 rpm and 45°C to form a uniform lipid ethanol solution, and then the lipid ethanol solution was evaporated under reduced pressure; the above steps were repeated 3 times to form a beaker VIII with a lipid film on the inner wall; 1 part of polyglycerol-10 stearate, 2 parts of sucrose stearate, 1 part of stearyl alcohol PEG-2, 3 parts of cetearyl glucoside, 5 parts of VC, and 1 part of gentianin were added to water and dissolved, and the mixture was stirred at 45°C for 30 minutes. The mixture was stirred with a magnetic stirrer at 0 rpm for 60 min to form a uniform solution VIII; the uniform solution VIII was added into beaker VIII, and magnetically stirred at 50°C and 300 rpm for 30 min to form a uniform mixed solution VIII; the mixed solution VIII was subjected to high-speed shearing at room temperature for 15 min, and the shear rate was 12000 rpm to obtain liposome colostrum VIII; the liposome colostrum VIII was homogenized twice under a high-pressure homogenizer at a homogenization pressure of 1200 bar and a temperature of 45°C, and then cooled to room temperature to obtain liposome VIII.

[0039] Example 2

[0040] This example provides a liposome stability test experiment.

[0041] The object to be tested is placed in a sealed container, placed at a specific temperature for a specific period of time, and the particle size of the object to be tested is checked at a specific time point. In this example, liposomes I to VIII of Example 1 are used as the objects to be tested, respectively, and placed at -18°C, 4°C, room temperature and 45°C for 360 days. At the initial stage, 90 days, 180 days and 360 days, after being diluted to a certain concentration with deionized water at room temperature, the average particle size of the liposomes is measured using dynamic light scattering technology, and the results shown in Table 1 are obtained.

[0042] Table 1 Sample particle size stability test results (unit: nm)

[0043]

[0044] As shown in Table 1, except for liposome IV-a, liposomes I to VIII all showed good particle size stability; the longer the storage time, the more significant the particle size stability.

[0045] Example 3

[0046] This example provides an experiment to verify the application performance of liposomes.

[0047] 1. ABTS free radical scavenging experiment

[0048] Mix 10 μL of the test object with 190 μL of ABTS working solution / PBS with a concentration of 0.25 mmol / L, shake well to obtain the ABTS working solution solution of the test object and the PBS solution of the test object, incubate at room temperature for 6 minutes, and measure the absorbance at 405 nm. The absorbance of the ABTS working solution of the test object is recorded as A1, the absorbance of the PBS solution of the test object is recorded as A2, and the absorbance of the blank ABTS working solution and blank PBS is recorded as A3. According to the formula Calculate the ABTS free radical scavenging rate. In this example, 100 mg / mL VC (ABTS free radical scavenging rate is 100%) is used as a positive control, and liposomes I to VIII of Example 1 are used as test substances, respectively. Figure 2 And the results shown in Table 2, where Figure 1 This is the standard curve of vitamin C (VC) scavenging rate of ABTS free radicals.

[0049] Table 2 ABTS free radical scavenging rate results

[0050]

[0051] When the positive control group IC 50 When the concentration is between 0.040 and 0.070 g / L, the system is considered effective. Figure 1 The IC of the positive control VC is 50 The test system is effective within the range of 40 μg / mL to 70 μg / mL. Figure 2 It can be seen that, except for liposome IV-a, the other liposomes have a high ABTS free radical scavenging rate, among which liposome IV has the highest average of 67.34%, indicating that the liposomes provided by the present invention have a synergistic effect in the scavenging effect of the active substance on ABTS free radicals.

[0052] 2. Antioxidant experiment of HaCaT cells

[0053] The test substance was diluted with DMEM complete medium to a final concentration of 2% as the test sample; H2O2 was diluted with DMEM complete medium to 800 μM as the control sample. HaCaT cells (human immortalized keratinocytes) in the logarithmic growth phase were cultured at 8×10 3Cells were inoculated into 96-well plates, with 100 μL of the system in each well. The outermost circle of the 96-well plate was sealed with 100 μL of PBS. After 24 hours, the cells were completely attached to the wall and replaced with the sample to be tested. The cells were cultured for another 24 hours. The supernatant was discarded. Except for the normal group, the other groups were given control samples. After 4 hours of culture in the incubator, the cell activity was detected using the CCK8 method immediately after modeling. The detection wavelength was 450 nm. In this example, 100 mg / mL of VC (ABTS free radical scavenging rate was 100%) was used as a positive control, and liposomes I to VIII of Example 1 were used as the test substances, respectively. The following results were obtained: Figure 3 Results shown.

[0054] Depend on Figure 3 It can be seen that after H2O2 injury, the cell viability of the group without drug administration was only 36.58%, indicating that the H2O2 injury HaCaT model was successfully established. Compared with the H2O2 group, liposome I to liposome VIII groups can improve the viability of oxidatively damaged HaCaT cells, and the difference is statistically significant (P < 0.0001); the effect of liposome IV-a group and VC group on improving the viability of oxidatively damaged HaCaT cells is not significant; among them, liposome IV has the strongest antioxidant damage performance, indicating that the liposomes provided by the present invention have a synergistic effect in exerting antioxidant effects.

[0055] 3. RAW264.7 cell anti-inflammatory experiment

[0056] RAW264.7 cells in the logarithmic growth phase were inoculated in a 24-well plate at a cell density of 5 × 10 5 / mL, 1mL per well. After incubation for 24 hours, the supernatant was aspirated and 1mL of the following samples were added respectively: ① Normal group: DMEM complete medium (5% FBS; 1% double antibody); ② LPS group: 1μg / mL LPS; ③ Experimental group: test substance (1%) + LPS (1μg / mL) (all samples were diluted with DMEM complete medium). After acting on the cells for 24 hours, the supernatant was aspirated into a centrifuge tube with a pipette, centrifuged at 1000r / min for 20min, the precipitate was discarded and the supernatant was aspirated. The change in the amount of inflammatory factors in the supernatant was detected using a TNF-α inflammatory factor detection kit, and the detection was performed according to the instructions of the kit. In this example, 100mg / mL of VC (ABTS free radical scavenging rate is 100%) was used as a positive control, and liposomes I to liposomes VIII in Example 1 were used as test substances, respectively, to obtain the following results. Figure 4 Results shown.

[0057] Depend on Figure 4It can be seen that the TNF-α level in the LPS group was significantly higher than that in the normal group, indicating that the inflammatory induction model was successfully established. Liposome I to liposome VIII groups can significantly reduce the level of inflammatory factor TNF-α in RAW 264.7 cells, and the difference is statistically significant (P < 0.01); the liposome IV-a group and VC group have no significant effect on inhibiting the inflammatory factor TNF-α; among them, liposome IV has the best effect on inhibiting the inflammatory factor TNF-α, indicating that the liposomes provided by the present invention have a synergistic effect in exerting anti-inflammatory effects.

[0058] Example 4

[0059] This example provides a melanin inhibition experiment using vitamin C liposomes.

[0060] 1. Tyrosinase activity detection experiment

[0061] Add 80 μL PBS buffer, 40 μL sample diluent of different concentrations, 40 μL tyrosinase solution and 80 μL substrate L-tyrosine solution to a 96-well plate, mix well and incubate at 25°C in the dark for 20 minutes. The absorbance was then detected at 475 nm on an ELISA reader to calculate the effect of the sample on the activity of elastase and the statistical difference P value. The absorbance of the negative and reaction groups was recorded as A0, the absorbance of the negative blank group was recorded as A1, the absorbance of the sample and reaction group was recorded as B0, and the absorbance of the sample and blank group was recorded as B1. The positive control sample substrate was used as the sample solution for reaction control, and the negative control group, i.e., the sample diluent, was used as the sample solution for reaction control. The tyrosinase inhibition rate was calculated as follows: Tyrosinase In this example, 100 mg / mL VC (ABTS free radical scavenging rate is 100%) was used as a positive control, and liposomes I to VIII of Example 1 were used as test samples, respectively, to obtain the results shown in Table 3. The experimental data were all expressed as mean ± standard deviation, and GraphpadPrism 8 software was used for plotting and data processing. AVONA was used for comparative analysis between different treatment groups, and the results were as follows: Figure 5 Results shown.

[0062] Table 3 Tyrosinase inhibition rate

[0063]

[0064]

[0065] Note: The inhibition rate / clearance rate results are rounded to two decimal places. The positive control concentration in the above table is 80 μg / mL. When p<0.05, it is significant.

[0066] From Table 3 and Figure 5It can be seen that liposomes I to VIII can significantly inhibit tyrosinase, and the difference is statistically significant (P < 0.05); the liposome IV-a group and VC group have no significant effect on inhibiting tyrosinase, indicating that the liposomes provided by the present invention have a synergistic effect in improving the inhibitory effect of tyrosinase.

[0067] Example 5

[0068] This example provides an application experiment of an antioxidant preparation with vitamin C liposome as an active ingredient.

[0069] 20 parts of the liposome to be tested, 10 parts of 1% sodium hyaluronate, and 10 parts of glycerol were added to 10 parts of purified water and stirred at 50°C and 500 rpm for 30 minutes to form a uniform mixed solution a; 10 parts of water, 10 parts of glycerol, 2 parts of soybean lecithin, and 5 parts of hydrogenated lecithin were added to a beaker and sheared under a shearing machine at a shearing speed of 10,000 rpm and a shearing time of 3 minutes to form a uniform mixed emulsion b; the mixed solution a and the mixed solution b were stirred at 37°C and 500 rpm for 30 minutes to be evenly mixed to obtain an antioxidant milk.

[0070] The liposome IV, liposome IV-a and VC with a concentration of 10% in Example 1 were used as the liposomes to be tested to prepare antioxidant milk IV, antioxidant milk IV-a and antioxidant milk VC, respectively, and the specific steps were as follows:

[0071] The obtained antioxidant milk was subjected to an effect verification experiment, and the steps are as follows:

[0072] A total of 90 volunteers with similar skin conditions, good health, and no history of cosmetic allergies were selected as subjects and randomly divided into 3 groups (antioxidant milk IV group, antioxidant milk IV-a group, and antioxidant milk VC group). The subjects used antioxidant milk twice a day, 1 mL each time, evenly applied to the face, for 28 consecutive days. The Visia skin detector was used to detect the number of skin pores, melanin and spot changes, skin color uniformity, and other data of the subjects. The results are shown in Table 4. According to the formula: Calculate the improvement rate and record the results in Table 4 and Figure 6 middle.

[0073] Table 4 Skin detection mean value change table (mm 2 )

[0074]

[0075] From Table 4 and Figure 6It can be seen that after using the antioxidant milk prepared with liposome IV as the main active ingredient, the pores, black spots, and skin color data of the subjects' facial skin were improved, the pores were reduced, the melanin and spots were reduced, and the skin color became whiter; the improvement effects of the other two groups were not significant, indicating that the prepared antioxidant milk has antioxidant and whitening effects, and has a synergistic effect.

[0076] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0077] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A vitamin C liposome, characterized in that: The total weight is 100 parts, including the following components: 1 to 20 parts of emulsifier, 1 to 10 parts of lipid, 1 to 10 parts of vitamin C, 1 to 4 parts of compound functional substance, and the rest is water; Wherein, the compounded effective substance is any one of ginsenoside Rg3, ginsenoside Rh2 and gentianin or a combination of several of them.

2. The vitamin C liposome according to claim 1, characterized in that The invention comprises the following components: 5 to 15 parts of emulsifier, 1 to 8 parts of lipid, 2 to 8 parts of vitamin C, 1 to 4 parts of compound functional substance, and the rest is water.

3. The vitamin C liposome according to claim 1, characterized in that It includes the following components: 5 parts of emulsifier, 4 parts of lipid, 4 parts of vitamin C, 3 parts of compound functional ingredients, and the rest is water.

4. The vitamin C liposome according to claim 1, characterized in that The emulsifier is any one or a combination of polyglyceryl-10 stearate, PEG-8 caprylic / capric glyceride, sucrose stearate, cetearyl glucoside, stearyl alcohol PEG-2, and polysorbate-80; The lipid is any one of cholesterol, soybean lecithin, hydrogenated lecithin, egg yolk lecithin, phosphatidylcholine, and phosphatidylserine, or a combination of several of them.

5. The method for preparing the vitamin C liposome according to any one of claims 1 to 4, characterized in that: The following steps are involved: dissolving the lipid in ethanol to obtain an ethanol solution of the lipid, and evaporating the ethanol solution of the lipid under reduced pressure to form a thin film on the inner wall of the container; Repeat the above step 1 to 3 times in the same container to obtain a container with a lipid film attached to the inner wall; The emulsifier, the vitamin C and the compound functional substance are uniformly dissolved in water at 30-50° C. to obtain a mixed solution; Adding the mixed solution into the container with the lipid film attached to the inner wall, and stirring to obtain a uniform mixed solution; Shearing the uniform mixed solution at room temperature to obtain liposome colostrum; The liposome colostrum is homogenized several times under a high-pressure homogenizer and cooled to room temperature to obtain the vitamin C liposome.

6. The preparation method according to claim 5, characterized in that: The uniform dissolution in water is carried out under the following conditions: stirring at a speed of 100 to 500 rpm for 30 to 60 minutes; And / or, the stirring to obtain a uniform mixed solution is carried out under the following conditions: stirring at a rotation speed of 100 to 500 rpm for 30 to 60 min; And / or, the shearing is carried out under the conditions of: shearing at a rotation speed of 8000 to 12000 rpm for 2 to 5 minutes; And / or, the homogenization conditions are: pressure 500-1200 bar and temperature 40-60°C.

7. Use of the vitamin C liposome according to any one of claims 1 to 4 or the vitamin C liposome prepared by the preparation method according to any one of claims 5 to 6 in the preparation of an antioxidant preparation.

8. The use according to claim 7, characterized in that: The antioxidant preparation is prepared into any one of a solution, a lotion, an ointment, a powder, a pill, a tablet, a film, an ointment, a gel, a suppository, a paste, an aerosol, and a spray.

9. An antioxidant preparation, characterized in that The invention comprises the following components: the vitamin C liposome described in any one of claims 1 to 4 or the vitamin C liposome prepared by the preparation method described in any one of claims 5 to 6 and auxiliary materials.

10. The antioxidant preparation according to claim 9, characterized in that The raw materials are calculated by weight, and the preparation method comprises the following steps: 20 parts of the vitamin C liposomes, 10 parts of 1% sodium hyaluronate, 10 parts of glycerol, and 20 parts of water were stirred at 50° C. and 500 rpm for 30 minutes to form a uniform mixed solution a; 10 parts of water, 10 parts of glycerol, 2 parts of soybean lecithin and 5 parts of hydrogenated lecithin were mixed, and sheared under a shearing machine at a shearing speed of 10000 rpm and a shearing time of 3 minutes to form a uniform mixed emulsion b; The mixed solution a and the mixed emulsion b were stirred at 37° C. and 500 rpm for 30 min to be uniformly mixed, thereby obtaining the antioxidant preparation.

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