Phenolic acid nanosom and preparation method and application thereof

Abstract

This invention discloses a phenolic acid nano-alcoholic vesicle, its preparation method, and its application. This phenolic acid nano-alcoholic vesicle comprises biocompatible lipids, nonionic surfactants, medium-chain diols, transdermal absorption enhancers, phenolic acid, and water. Based on biocompatible lipids as the vesicle wall material, this invention employs nonionic surfactants as stabilizers to improve the stability, encapsulation efficiency, and penetration enhancement of the vesicles. The use of medium-chain diols facilitates the dissolution of phenolic acid, the formation of uniform vesicle vesicles, improves vesicle stability, and promotes transdermal absorption. The use of transdermal absorption enhancers regulates the surface properties of the vesicles, significantly enhancing the transdermal permeation, skin retention, and bioavailability of the active ingredients encapsulated in the vesicles, thereby greatly reducing the amount of active ingredients used and the raw material cost of cosmetics.

Description

Technical Field

[0001] This invention relates to the field of cosmetics, and more particularly to a phenolic acid nano-olite, its preparation method, and its application. Background Technology

[0002] Biological oxidation is a series of redox reactions in which substances are oxidized to produce energy during the body's metabolism. Skin cells, through metabolism or under external aggressors including sunlight, ionizing radiation, and pollutants, continuously produce highly reactive free radicals and reactive oxygen species (ROS). Excessive production of these highly reactive molecules can lead to changes in cell structure and premature cell death, causing skin problems such as skin aging, dullness, sensitivity, and skin cancer. Phenolic acids are a class of plant extracts with antioxidant activity. Most polyphenols exist in a conjugated structure, exhibiting excellent antioxidant activity and making them ideal antioxidants in cosmetics. Currently, ferulic acid, ellagic acid, salicylic acid, and caffeic acid are added as antioxidant active ingredients to creams, serums, and masks. However, these phenolic acid active ingredients generally have the following problems when added to cosmetics: (1) Poor stability: Phenolic acids have extremely poor light, heat, and pH stability. When added to cosmetics as active ingredients, the active ingredients are prone to change during emulsification and mixing with other formulation ingredients, as well as during the transportation, storage, and use of cosmetics, leading to a reduction or loss of cosmetic efficacy; (2) Low transdermal absorption efficiency: Due to the skin's own barrier, phenolic acid active ingredients have difficulty penetrating the stratum corneum and acting on skin targets, resulting in minimal skin care effects; (3) Easily causes skin irritation: Some phenolic acid active ingredients are irritating. If added directly to cosmetics, improper concentration control can easily cause skin irritation or even skin inflammation. Therefore, enhancing the stability of active ingredients and improving transdermal absorption and utilization are urgent technical problems that need to be solved when phenolic acid active ingredients are applied to cosmetics.

[0003] Alcoholic liposomes possess advantages such as good membrane fluidity, good transdermal absorption, high encapsulation efficiency, good stability, and the ability to bind lipophilic, hydrophilic, and amphiphilic drugs. They are also non-biotoxic carriers and are commonly used as carriers for active ingredients in cosmetics to improve the stability of active ingredients and promote their transdermal absorption and efficacy in the skin. However, in existing invention patents, CN110200829A and CN111617039A use volatile ethanol as an organic solvent to prepare resveratrol liposomes and curcumin liposomes. Due to the strong skin irritation and high volatility of ethanol, the prepared liposomes are unstable, and the potential skin irritation cannot be ignored. CN108992383A prepares a liposome of vine tea flavonoid composition, which does not use a nonionic surfactant, potentially leading to poor stability, easy aggregation or delamination, and low encapsulation efficiency. CN102397255A prepares a progesterone liposome, but the short-chain alcohol does not significantly improve the transdermal absorption efficiency of the encapsulated active ingredient. Summary of the Invention

[0004] To overcome the problems of poor stability and low transdermal absorption efficiency of existing phenolic acid nanoparticles, one objective of this invention is to provide a phenolic acid nanoparticle; another objective is to provide a method for preparing such a phenolic acid nanoparticle; and a third objective is to provide the application of such a phenolic acid nanoparticle in cosmetic preparation.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0006] The first aspect of the present invention provides a phenolic acid nano-ol body, the phenolic acid nano-ol body comprising a biocompatible lipid, a nonionic surfactant, a medium-chain diol, a transdermal absorption enhancer, phenolic acid, and water; wherein the carbon chain of the medium-chain diol in the present invention refers to C4-C12.

[0007] Preferably, this phenolic acid nanoparticle comprises the following components in parts by weight: 0.1-5 parts of biocompatible lipids, 0.05-3 parts of nonionic surfactants, 1-50 parts of medium-chain diols, 0.001-2 parts of transdermal absorption enhancers, 0.01-5 parts of phenolic acids, and 45-85 parts of water.

[0008] More preferably, this phenolic acid nanoparticle comprises the following components in parts by weight: 0.2-4.5 parts of biocompatible lipids, 0.1-2.5 parts of nonionic surfactants, 10-50 parts of medium-chain diols, 0.01-1 parts of transdermal absorption enhancers, 0.1-3 parts of phenolic acids, and 45-80 parts of water.

[0009] In a further preferred embodiment, this phenolic acid nano-ol body comprises the following components in parts by weight: 0.2-4 parts of biocompatible lipids, 0.2-2 parts of nonionic surfactants, 15-45 parts of medium-chain diols, 0.01-0.5 parts of transdermal absorption enhancers, 0.2-2 parts of phenolic acids, and 50-80 parts of water.

[0010] Preferably, the particle size of this phenolic acid nanoparticle is 50-150 nm; more preferably, the particle size of the phenolic acid nanoparticle is 50-120 nm.

[0011] Preferably, the polydispersity of this phenolic acid nano-alcohol is <0.1; more preferably, the polydispersity of the phenolic acid nano-alcohol is <0.07.

[0012] Preferably, the encapsulation efficiency of this phenolic acid nano-alcohol body is >80%.

[0013] Preferably, the biocompatible lipids in this phenolic acid nanoparticle include at least one of soybean lecithin, hydrogenated soybean lecithin, egg yolk lecithin, phosphatidylcholine, hydrogenated phosphatidylcholine, phosphatidylinositol, phosphatidylethanolamine, and cholesterol.

[0014] Preferably, in this phenolic acid nanoparticle, the nonionic surfactant includes at least one of Tween, Span, and poloxamer.

[0015] Preferably, in this phenolic acid nano-alcoholic granule, the medium-chain diol includes at least one of butanediol, pentanediol, hexanediol, heptanediol, octanediol, nonanediol, decanediol, undecanediol, and dodecanediol. The medium-chain diol of the present invention can promote the dissolution of phenolic acids, act as a co-emulsifier, and promote the formation of uniform alcoholic vesicles. Compared with short-chain alcohol solvents commonly used in alcoholic granules, such as ethanol, propylene glycol, and isopropanol, it can improve the stability of alcoholic granules and has a permeation-promoting effect.

[0016] Preferably, in this phenolic acid nanoparticle, the transdermal absorption enhancer is a polysaccharide derivative; more preferably, the transdermal absorption enhancer includes at least one of sucrose laurate, sucrose monopalmitate, sucrose dipalmitate, saponins, monophospholipid A, and dioleoylphosphatidylethanolamine-hyaluronic acid.

[0017] Preferably, in this phenolic acid nanoparticle, the phenolic acid includes hydroxybenzoic acid, hydroxycinnamic acid and their derivatives; more preferably, the phenolic acid includes at least one of ferulic acid, ellagic acid, salicylic acid, caffeic acid, protocatechuic acid, gallic acid, sinapic acid, cinnamic acid, chlorogenic acid, vanillic acid, p-coumaric acid, styric acid, p-hydroxybenzoic acid, syringic acid, gentianic acid, epicatechin, oleanolic acid, gambogeylic acid, ursolic acid, cinnamic acid, salvianolic acid, rosmarinic acid and sarsaparilla acid.

[0018] A second aspect of the present invention provides a method for preparing the above-mentioned phenolic acid nano-olite, comprising the following steps:

[0019] (1) A mixture of biocompatible lipids, nonionic surfactants, medium-chain diols, dermal absorption enhancers and phenolic acids was prepared to obtain an alcohol phase;

[0020] (2) The alcohol phase is mixed with water and homogenized to obtain the phenolic acid nano-alcohol body.

[0021] Preferably, in the preparation method of this phenolic acid nano-alcohol body, in step (2), the alcohol phase and water are mixed by at least one of syringe injection, microchannel reactor mixing, and ultrasonic microreactor mixing.

[0022] More preferably, the injection flow rate of the syringe is 0.1-5 mL / min; even more preferably, the injection flow rate of the syringe is 0.5-2 mL / min.

[0023] More preferably, the flow rate of the alcohol phase mixed in the microchannel reactor is 0.1-5 mL / min; even more preferably, the flow rate of the alcohol phase mixed in the microchannel reactor is 0.5-2 mL / min.

[0024] More preferably, the flow rate of the aqueous phase mixed in the microchannel reactor is 0.3-15 mL / min; even more preferably, the flow rate of the aqueous phase mixed in the microchannel reactor is 1.5-6 mL / min.

[0025] More preferably, the flow rate of the alcohol phase mixed in the ultrasonic microreactor is 0.1-5 mL / min; even more preferably, the flow rate of the alcohol phase mixed in the ultrasonic microreactor is 0.5-2 mL / min.

[0026] More preferably, the flow rate of the aqueous phase mixed in the ultrasonic microreactor is 0.3-15 mL / min; even more preferably, the flow rate of the aqueous phase mixed in the ultrasonic microreactor is 1.5-6 mL / min.

[0027] More preferably, the ultrasonic power of the ultrasonic microreactor mixing is 10-60W; even more preferably, the ultrasonic power of the ultrasonic microreactor mixing is 15-30W.

[0028] Preferably, in the preparation method of this phenolic acid nano-alcohol body, in step (2), the homogenization is carried out by at least one of shear homogenization, ultrasonic homogenization, high pressure homogenization and microfluidic homogenization.

[0029] More preferably, the shearing rate for homogenization is 3000-25000 rpm; even more preferably, the shearing rate is 5000-15000 rpm.

[0030] Further preferably, the ultrasonic power for ultrasonic homogenization is 150-500W; even more preferably, the ultrasonic power is 200-300W.

[0031] More preferably, the pressure of high-pressure homogenization is 15,000-30,000 Psi; even more preferably, the pressure of high-pressure homogenization is 18,000-25,000 Psi.

[0032] More preferably, the pressure for microjet homogenization is 15,000-30,000 Psi; even more preferably, the pressure for microjet homogenization is 18,000-25,000 Psi.

[0033] Further preferably, the number of cycles for high-pressure homogenization is 1-6; even more preferably, the number of cycles for high-pressure homogenization is 1-3.

[0034] More preferably, the number of cycles of microfluidic treatment is 1-6; even more preferably, the number of cycles of microfluidic treatment is 1-3.

[0035] Preferably, in the preparation method of this phenolic acid nano-olite, the reaction temperature of the mixing step in step (1) is 15-80℃; more preferably, the reaction temperature of the mixing step is 30-80℃.

[0036] The third aspect of this invention provides the application of the above-mentioned phenolic acid nano-olite in the preparation of cosmetics.

[0037] Preferably, the above-mentioned phenolic acid nano-olite is applied to cosmetics, wherein the cosmetics include at least one of creams, lotions, oils, toners, eye care products, masks, and facial cleansers.

[0038] The beneficial effects of this invention are:

[0039] This invention utilizes biocompatible lipids as the wall material for liposomes and employs nonionic surfactants as stabilizers to improve the stability, encapsulation efficiency, and penetration enhancement of liposomes. The use of medium-chain diols facilitates the dissolution of phenolic acids, forms uniform liposome vesicles, improves liposome stability, and promotes transdermal absorption. The use of transdermal absorption enhancers regulates the surface properties of the liposome vesicles, significantly enhancing the transdermal permeation, skin retention, and bioavailability of the encapsulated active ingredients, thereby greatly reducing the amount of active ingredients used and the raw material cost of cosmetics. The phenolic acid nano-liposomes of this invention overcome the shortcomings of poor light and heat stability of phenolic acid active ingredients, resulting in stable liposomes with high encapsulation efficiency and strong transdermal absorption. When applied to cosmetics, they can significantly enhance the efficacy of phenolic acid active ingredients.

[0040] The small particle size and high surface activity of the liposomes in this invention can promote the penetration of phenolic acid through the stratum corneum; furthermore, the surface of the transdermal absorption promoter has moderate hydrophilicity and hydrophobicity, and is relatively hydrophilic, so that after the phenolic acid penetrates the stratum corneum, it is more easily retained by the epidermis and dermis.

[0041] The phenolic acid nanoparticles in this invention exhibit significantly increased skin permeability and skin retention over 24 hours, ranging from 201 to 499 μg / cm³. 2 and 63-90 μg / cm 2 The 24-hour skin penetration and retention of these compounds are approximately 5-7.9 times and 3.4-6.1 times that of a simple mixture of phenolic acid, nonionic surfactant, medium-chain diol, and water, respectively. They are also approximately 2.1-3.4 times and 1.5-2.2 times that of alcoholic compounds without added transdermal absorption enhancers, respectively. This overcomes the poor light and heat stability of phenolic acid active ingredients, resulting in stable alcoholic compounds with high encapsulation efficiency and strong transdermal absorption. When applied in cosmetics, these compounds can significantly enhance the efficacy of phenolic acid active ingredients.

[0042] The phenolic acid proteosome of the present invention can provide high phenolic acid solubility, a wide range of active ingredient loading concentration (0.01-5%), nano-sized and uniformly distributed particles with a particle size of 50-150 nm, a PDI of less than 0.1, and an encapsulation efficiency of greater than 80%. Detailed Implementation

[0043] The following will describe the concept and technical effects of the present invention clearly and completely with reference to embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.

[0044] The particle size and polydispersity of the phenolic acid nanoparticles in the examples and comparative examples were tested using a Zetasizer NanoZS90 nanoparticle size potentiometer, and the encapsulation efficiency of the phenolic acid nanoparticles was determined using an ultrafiltration tube. The particle size, polydispersity, and encapsulation efficiency data of the phenolic acid nanoparticles were all tested using existing methods.

[0045] In the following examples and comparative examples, "parts" refers to "parts by mass"; the water balance in the following examples refers to the balance after other components when the total mass is 100 parts.

[0046] Example 1

[0047] A transdermal phenolic acid nanoparticle comprising 1.5 parts soybean lecithin, 0.5 parts Tween 80, 35 parts pentylene glycol, 0.1 parts sucrose laurate, 0.75 parts ferulic acid, and the balance being water.

[0048] The phenolic acid nano-olite was prepared using the following method:

[0049] Soybean lecithin, Tween 80, pentylene glycol, sucrose laurate, and ferulic acid were mixed at 35°C to prepare an alcohol phase. Water was heated to 35°C to prepare an aqueous phase. The alcohol phase was injected into the aqueous phase using a syringe at a rate of 1 mL / min while stirring to ensure uniform mixing, thus initially preparing phenolic acid nanoparticles. The initially prepared phenolic acid nanoparticles were subjected to microfluidic treatment at a pressure of 20,000 Psi for one cycle to obtain phenolic acid nanoparticles. The particle size of the phenolic acid nanoparticles was 83.4 nm, the polydispersity was 0.040, and the encapsulation efficiency was 92.89%.

[0050] Example 2

[0051] A transdermal phenolic acid nanoparticle comprising 2.2 parts egg yolk lecithin, 0.5 parts Tween 20, 20 parts hexanediol, 0.25 parts sucrose monopalmitate, 1.2 parts ellagic acid, and the balance being water.

[0052] The phenolic acid nano-olite was prepared using the following method:

[0053] Egg yolk lecithin, Tween 20, hexanediol, sucrose monopalmitate, and ellagic acid were mixed at 35°C to prepare an alcohol phase. Water was heated to 35°C to prepare an aqueous phase. The alcohol phase was injected into the aqueous phase at a rate of 1 mL / min using a syringe while stirring to ensure homogeneity, thus initially preparing phenolic acid nanoparticles. The initially prepared phenolic acid nanoparticles were subjected to microfluidic treatment at a pressure of 20,000 Psi for one cycle to obtain phenolic acid nanoparticles. The particle size of the phenolic acid nanoparticles was 110.1 nm, the polydispersity was 0.036, and the encapsulation efficiency was 85.43%.

[0054] Example 3

[0055] A transdermal phenolic acid nanoparticle comprising 0.5 parts phosphatidylcholine, 0.15 parts Span 20, 30 parts octanediol, 0.15 parts sucrose dipalmitate, 0.5 parts protocatechuic acid, and the balance being water.

[0056] The phenolic acid nano-olite was prepared using the following method:

[0057] Phosphatidylcholine, Span 20, octanediol, sucrose dipalmitate, and protocatechuic acid were mixed at 60°C to prepare an alcohol phase. Water was heated to 60°C to prepare an aqueous phase. The alcohol phase and aqueous phase were injected into the channels of a microreactor at rates of 1 mL / min and 3 mL / min, respectively, to initially prepare phenolic acid nanoparticles. The initially prepared phenolic acid nanoparticles were subjected to shear homogenization at a shear rate of 5000 rpm to obtain phenolic acid nanoparticles. The particle size of the phenolic acid nanoparticles was 93.6 nm, the polydispersity was 0.020, and the encapsulation efficiency was 96.01%.

[0058] Example 4

[0059] A transdermal phenolic acid nanoparticle comprising 1.5 parts of hydrogenated phosphatidylcholine, 0.5 parts of poloxamer 188, 20 parts of butylene glycol, 5 parts of undecanediol, 0.02 parts of saponins, 1.0 part of chlorogenic acid, and the balance being water.

[0060] The phenolic acid nano-olite was prepared using the following method:

[0061] Hydrogenated phosphatidylcholine, poloxamer 188, butylene glycol, undecanediol, saponins, and chlorogenic acid were mixed at 75°C to prepare an alcohol phase. Water was heated to 75°C to prepare an aqueous phase. The alcohol phase and aqueous phase were injected into the channels of an ultrasonic microreactor at rates of 1 mL / min and 3 mL / min, respectively, with an ultrasonic power of 30 W, to initially prepare phenolic acid nanoparticles. The initially prepared phenolic acid nanoparticles were subjected to high-pressure homogenization at a pressure of 25000 Psi for 2 cycles to obtain phenolic acid nanoparticles. The particle size of the phenolic acid nanoparticles was 66.2 nm, the polydispersity was 0.032, and the encapsulation efficiency was 89.10%.

[0062] Example 5

[0063] A transdermal phenolic acid nanoparticle comprising 3.0 parts soybean lecithin, 0.6 parts cholesterol, 1.6 parts Tween 40, 35 parts butylene glycol, 5 parts decanediol, 0.035 parts monophospholipid A, 1.5 parts gentian acid, and the balance water.

[0064] The phenolic acid nano-olite was prepared using the following method:

[0065] Soybean lecithin, cholesterol, Tween 40, butylene glycol, decanediol, monophospholipid A, and gentic acid were mixed at 30°C to obtain an alcohol phase. Water was heated to 30°C to obtain an aqueous phase. The alcohol phase was injected into the aqueous phase at a rate of 1 mL / min using a syringe while stirring to ensure homogeneity, thus initially preparing phenolic acid nanoparticles. The initially prepared phenolic acid nanoparticles were then subjected to ultrasonic homogenization at a power of 200 W to obtain phenolic acid nanoparticles. The particle size of these phenolic acid nanoparticles was 103.2 nm, the polydispersity was 0.066, and the encapsulation efficiency was 80.85%.

[0066] The above-mentioned phenolic acid nano-ol body is mixed evenly with the toner to obtain a toner containing phenolic acid nano-ol body.

[0067] Example 6

[0068] A transdermal phenolic acid nanoparticle comprising 0.7 parts phosphatidylethanolamine, 0.3 parts cholesterol, 0.3 parts poloxamer 407, 25 parts heptanediol, 0.04 parts dioleoylphosphatidylethanolamine-hyaluronic acid, 0.25 parts salvianolic acid, and the balance being water.

[0069] The phenolic acid nano-olite was prepared using the following method:

[0070] Phosphatidylethanolamine, cholesterol, poloxamer 407, heptanediol, dioleoylphosphatidylethanolamine-hyaluronic acid, and salvianolic acid were mixed at 65°C to prepare an alcohol phase. Water was heated to 65°C to prepare an aqueous phase. The alcohol phase was injected into the aqueous phase at a rate of 1 mL / min using a syringe injection method, while stirring to ensure uniform mixing, thus initially preparing phenolic acid nanoparticles. The initially prepared phenolic acid nanoparticles were subjected to microfluidic treatment at a pressure of 18000 Psi for one cycle to obtain phenolic acid nanoparticles. The particle size of the phenolic acid nanoparticles was 99.2 nm, the polydispersity was 0.010, and the encapsulation efficiency was 99.31%.

[0071] The above-mentioned phenolic acid nano-ol body was mixed evenly with the skin care lotion at room temperature to obtain a skin care lotion containing phenolic acid nano-ol body.

[0072] Comparative Examples 1-6

[0073] Compared with Examples 1-6, Comparative Examples 1-6 were identical in terms of ingredients and operating procedures, except that no biocompatible lipid components and transdermal absorption enhancers were added to the formulations.

[0074] Comparative Examples 7-12

[0075] Comparative Examples 7-12 are identical to Examples 1-6 except that no transdermal absorption enhancers were added to the formulations. All other components and operating procedures are the same.

[0076] Stability test

[0077] The phenolic acid nanoparticles prepared in Examples 1-6 were stored in sealed containers under the following conditions for 3 months: 4°C in the dark, 25°C in the dark, and 25°C without light protection. The properties, particle size, polydispersity, and encapsulation efficiency of the samples were then examined. The test results are shown in Table 1.

[0078] Table 1. Stability test results of phenolic acid nano-olite

[0079]

[0080]

[0081] As can be seen from Table 1, the phenolic acid nano-alcohol bodies provided by the present invention showed no agglomeration, discoloration, or stratification after being placed for 3 months. The particle size was between 50-120 nm, the polydispersity was <0.12, and the encapsulation rate was above 60%, which meets the requirements of practical applications.

[0082] Skin absorption and permeation performance test

[0083] The phenolic acid nanoparticles prepared in Examples 1-6 and Comparative Examples 1-12 were evaluated for skin absorption and permeability.

[0084] Test conditions: The in vitro transdermal experiment used a Franz diffusion cell, with the hairless abdominal skin of a one-month-old Bama miniature pig as the model. The diffusion cell parameters were: effective diffusion area 3.14 cm². 2 The receiving pool has a volume of 7.0 mL, the receiving solution is physiological saline, and the magnetic stirring speed is 200 rpm.

[0085] The specific testing method involves fixing pigskin between a receiving pool and a supply pool (with the inner skin facing the receiving pool). The receiving pool is maintained at a constant temperature of 37.0±0.5℃. At 1h, 2h, 4h, 6h, 8h, 10h, 12h, and 24h, 0.5mL of the receiving solution is collected, and 0.5mL of the analyte is added. The receiving solution is filtered through a 0.22μm organic membrane. The concentration of the active ingredient in the receiving solution is determined using high-performance liquid chromatography (HPLC). The cumulative transdermal permeation at each time point is calculated, and a transdermal absorption drug-time curve is plotted to understand the in vitro transdermal permeation effect of the active ingredient. The transdermal permeation rate per unit area of ​​the active ingredient is calculated using the following formula:

[0086]

[0087] Among them, Q s S is the cumulative transdermal dose; S is the effective diffusion area; V is the volume of physiological saline in the receiving tank; C i The concentration of the active ingredient in the receiving solution from the first to the last sampling is denoted as C; n is the volume of the nth sampling; C is the concentration of the active ingredient in the receiving solution from the first to the last sampling.n This represents the concentration of the active ingredient in the receiving solution during this sampling.

[0088] Table 2. Results of skin absorption and permeation performance tests of phenolic acid nanoparticles (24h, μg / cm³). 2 )

[0089]

[0090] As shown in Table 2, compared with Comparative Examples 1-6 and 7-12, the phenolic acid nano-alcohol bodies in Examples 1-6 showed a significant increase in skin penetration and retention over 24 hours. This indicates that the phenolic acid nano-alcohol body formulation and the added transdermal absorption enhancer can significantly enhance the penetration of the alcohol bodies into the skin and increase the retention of the phenolic acid active ingredients in the skin, thereby enabling the phenolic acid active ingredients to be rapidly delivered to the skin target sites where they take effect.

[0091] Cellular antioxidant activity and stability test

[0092] The phenolic acid nanoparticles prepared in Examples 1-6 and Comparative Examples 1-6 were evaluated for their cellular antioxidant activity. Furthermore, the cellular antioxidant activity of the above-mentioned phenolic acid nanoparticles was evaluated again after being stored at 25°C under light-free conditions for 3 months to examine the stability of the antioxidant activity of the phenolic acid active ingredients encapsulated in the nanoparticles.

[0093] The testing procedure is as follows: Human skin keratinocytes (HaCaT cells) in the logarithmic growth phase are taken and processed at a ratio of 2 × 10⁻⁶. 5 HaCaT cells were seeded per well in a 6-well plate and cultured for 24 h. Then, 100 μg / mL of the test sample (based on the concentration of active ingredient in the sample) was added. Each dose group was in 6 replicates. After pretreatment for 6 h, 400 μmol / L tert-butyl hydroperoxide (tBHP) was added and the treatment continued for another 6 h. HaCaT cells were collected by centrifugation, the culture medium was removed, and serum-free medium containing DCFH-DA (10 μmol / L) fluorescent probe was added. After thorough mixing, the cells were incubated at 37 °C in a 5% CO2 incubator. The cells were inverted and mixed every 5 min. After 20 min of culture, the cells were collected by centrifugation, washed twice with PBS, and resuspended in 500 μL of PBS. The fluorescence intensity was detected by flow cytometry.

[0094] Table 3 Results of the cellular antioxidant effects of phenolic acid nanoparticles

[0095]

[0096] As shown in Table 3, compared with Comparative Examples 1-6, the phenolic acid nanoparticles in Examples 1-6 can effectively reduce intracellular ROS levels and exert the beneficial antioxidant effects of phenolic acid active ingredients. More importantly, the antioxidant effects of the phenolic acid nanoparticles in Examples 1-6 are more stable, indicating that phenolic acid nanoparticles can overcome the disadvantage of poor stability of phenolic acid active ingredients and significantly enhance the efficacy of phenolic acid active ingredients in cosmetic applications.

[0097] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A phenolic acid nano-alcoholic liposome, characterized in that, The phenolic acid nano-ol body is composed of the following components in parts by weight: 0.1-5 parts biocompatible lipids, 0.05-3 parts nonionic surfactants, 1-50 parts medium-chain diols, 0.001-2 parts transdermal absorption enhancers, 0.01-5 parts phenolic acids, and 45-85 parts water. The nonionic surfactant is at least one of Tween, Span, and poloxamer. The transdermal absorption enhancer is at least one of monophospholipid A and dioleoylphosphatidylethanolamine-hyaluronic acid; The medium-chain diol is at least one of butanediol, pentanediol, hexanediol, and heptahydrate. The phenolic acid is at least one of ferulic acid, ellagic acid, protocatechuic acid, chlorogenic acid, gentian acid, and salvianolic acid.

2. The phenolic acid nano-alcohol body according to claim 1, characterized in that, The particle size of the phenolic acid nanoparticles is 50-150 nm.

3. The phenolic acid nano-alcohol body according to claim 1, characterized in that, The polydispersity of the phenolic acid nano-olite is <0.

1.

4. The phenolic acid nano-alcoholic body according to any one of claims 1 to 3, characterized in that, The biocompatible lipids include at least one of the following: soybean lecithin, hydrogenated soybean lecithin, egg yolk lecithin, phosphatidylcholine, hydrogenated phosphatidylcholine, phosphatidylinositol, phosphatidylethanolamine, and cholesterol.

5. A method for preparing phenolic acid nano-alcoholic liposomes according to any one of claims 1 to 4, characterized in that, Includes the following steps: (1) A mixture of biocompatible lipids, nonionic surfactants, medium-chain diols, dermal absorption enhancers and phenolic acids was prepared to obtain an alcohol phase; (2) The alcohol phase is mixed with water and homogenized to obtain the phenolic acid nano-alcohol body.

6. The application of the phenolic acid nano-alcohol body according to any one of claims 1 to 4 in the preparation of cosmetics.

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