Ethosome encapsulating galla rhois extract, preparation method therefor, and functional cosmetic composition using same

Abstract

The present invention relates to a method for preparing an ethosome encapsulating a Galla Rhois extract, an ethosome encapsulating a Galla Rhois extract prepared by the method, and a functional cosmetic composition using same. The present invention provides an ethosome encapsulating a Galla Rhois extract, which is a natural material having various biological activities such as antioxidant, anti-inflammatory, and antifungal effects, as well as skin tone-improving and astringent effects, and a method for preparing same. In addition, the present invention establishes optimal conditions for preparing ethosomes containing a Galla Rhois extract, thereby providing conditions that ensure effective loading and stability of the Galla Rhois extract under various conditions such as concentration, temperature, and pH. Furthermore, when cosmetics are prepared using ethosomes encapsulating a Galla Rhois extract, the present invention provides the effect of rapid absorption deep into the skin and sustained maintenance of oil and moisture.

Description

Ethosome encapsulated with Gallnut extract, method for manufacturing the same, and functional cosmetic composition using the same

[0001] The present invention relates to a method for manufacturing an ethosome containing a gallnut extract, an ethosome containing a gallnut extract manufactured by this method, and a functional cosmetic composition using the same. More specifically, the invention relates to a method for manufacturing an ethosome containing a gallnut extract that can significantly contribute to increasing skin absorption rates and maintaining a healthy skin condition when applied to cosmetics by encapsulating an active ingredient extracted from a natural material, gallnut, into an ethosome, and an ethosome containing a gallnut extract manufactured by this method and a functional cosmetic composition using the same.

[0002]

[0003] Various delivery methods have been developed to effectively deliver active ingredients from natural cosmetic materials deep into the skin; among these, the method utilizing liposomes is considered one of the significant innovations in the cosmetics industry. Liposomes are microscopic spherical vesicles composed of a phospholipid bilayer and are known as drug delivery systems that effectively deliver drugs or active ingredients. Using liposomes as a delivery system enables effective ingredient delivery, thereby increasing the absorption rate of the ingredients.

[0004] However, as it was reported that liposomes do not maintain a spherical shape and penetrate deep into the skin, but rather rupture on the skin surface, causing slight permeability due to the destruction of the chemical lipid layer, a new transdermal drug delivery system was required that could carry drugs like liposomes while allowing the carrier to penetrate deep into the skin without damage. Consequently, improved transdermal drug delivery systems emerged, such as ethosomes, transfersomes, transethosomes, niosomes, and exosomes, which are modified forms of liposomes. In particular, ethosomes are evaluated as a more skin-friendly delivery system compared to liposomes and have been proven to possess the ability to effectively deliver active ingredients to the deep layers of the skin. For example, it has been reported that ethosomes containing minoxidil exhibit a drug delivery efficiency of over 65% compared to liposomes and possess the property of rapidly penetrating the dermis. Ethosomes, which are a modified form of liposomes, have a phospholipid bilayer structure containing a high concentration of ethanol and are used in the fields of cosmetics and pharmaceuticals to improve the skin permeability of active ingredients.

[0005] However, in the conventional process of manufacturing etosome, liposomes are prepared by dissolving components such as phospholipids in ethanol and then slowly adding water to produce etosome, but due to the problem of evaporation of the ethanol, it is difficult to produce liposomes or etosome in a uniform form, such as in terms of structure and stability.

[0006] Furthermore, there are currently limitations in using etosomes to penetrate active ingredients deep into the skin. In other words, if active ingredients remain only on the surface of the skin, their effectiveness is inevitably limited in reaching the dermis or deeper layers. This ultimately acts as a limiting factor in maximizing the efficacy of the medicinal ingredients, and therefore, the development of a new methodology to effectively deliver active ingredients deep into the skin is required.

[0007] Accordingly, the inventors have completed the present invention by developing a method for manufacturing an etosome containing a gallnut extract that has antioxidant activity, whitening and natural antioxidant effects, and by establishing optimal conditions to improve the delivery amount and depth of the active ingredient in terms of skin delivery efficiency.

[0008]

[0009] The present invention aims to provide an ethosome containing a natural product, a gallnut extract, a method for manufacturing the same, and a functional cosmetic composition using the same as a solution to the problems described above.

[0010]

[0011] The present invention, for achieving the above objectives, provides a method for manufacturing an ethosome containing a gallnut extract as a means of solving the problem.

[0012] The above manufacturing method is characterized by comprising the steps of: manufacturing a liposome (S100); extracting Galla Rhois with ethanol to produce an Galla Rhois ethanol extract (S200); and mixing the liposome with the Galla Rhois ethanol extract to produce an ethosome containing the Galla Rhois ethanol extract (S300).

[0013] The step (S100) of manufacturing the above liposome comprises: (A) a step of manufacturing a lipid film by dissolving a lipid in an organic solvent and then removing the organic solvent; (B) a step of manufacturing a liposome in an emulsion state by hydrating the lipid film with a hydration solution; and (C) a step of homogenizing the liposome.

[0014] In the step of preparing a lipid film by dissolving the above (A) lipid in an organic solvent and then removing the organic solvent, the concentration of the lipid may be dissolved in the organic solvent to be 10 to 50 g / L.

[0015] In the step (B) of preparing liposomes in an emulsion state by hydrating the lipid film with a hydration solution, the concentration of the hydration solution may be 230 to 325 mM.

[0016] In the step (S300) of preparing the ethosome, the mixing ratio of the liposome and the gallnut extract may be 4 to 7:1.

[0017]

[0018] And the present invention provides another means of solving the problem by providing an ethosome containing a gallnut extract characterized by being manufactured by the above method.

[0019] The above ethosome is characterized by being stable under weakly acidic conditions of pH 4.5 to 5.5.

[0020]

[0021] Meanwhile, the present invention provides a cosmetic composition characterized by including an ethosome containing a gallnut extract as another means of solving the problem.

[0022]

[0023] The present invention has the effect of providing an ethosome containing a natural product, a gallnut extract, which has various biological activities such as antioxidant, anti-inflammatory, and antifungal effects, as well as skin tone improvement and astringent action, and a method for manufacturing the same.

[0024] In addition, the present invention has the effect of establishing optimal conditions for producing an ethosome containing gallnut extract, thereby providing conditions that ensure effective loading and stability of the gallnut extract under various concentrations, temperatures, pH, etc.

[0025] In addition, when a cosmetic product is manufactured using an ethosome containing a gallnut extract, the present invention has the effect of being rapidly absorbed deep into the skin, providing a continuous oil and moisture retention effect.

[0026] The effects of the present invention are not limited to those mentioned above, and other unmentioned effects will be clearly understood by those skilled in the art from the description in the claims.

[0027]

[0028] FIG. 1 is a process block diagram illustrating a method for manufacturing an ethosome containing a gallnut extract according to a preferred embodiment of the present invention.

[0029] FIG. 2 is a photograph illustrating the step of manufacturing a liposome according to a preferred embodiment of the present invention.

[0030] Figure 3 is a photograph of the hydration results of a lipid film taken with an optical microscope or a digital microscope.

[0031] Figure 4 is a photograph of liposome changes according to the homogenization method taken with an optical microscope (200x).

[0032] Figure 5 is a photograph of changes in liposomes according to storage temperature taken with an optical microscope (200x magnification).

[0033] Figure 6 is a photograph illustrating the steps for preparing an obezza ethanol extract according to a preferred embodiment of the present invention.

[0034] FIG. 7 is a photograph illustrating the step of preparing an ethosome containing a gallnut ethanol extract according to a preferred embodiment of the present invention.

[0035] Figure 8 is a photograph of gallnut-ethosome taken with an optical microscope (200x) according to mixing time.

[0036] Figure 9 is a photograph of gallnut-ethosome taken with an optical microscope (200x) according to mixing temperature.

[0037] Figure 10 is a photograph of gallnut-ethosome taken with an optical microscope (200x) with or without filtering.

[0038] Figure 11 is a graph showing the liposome yield according to osmotic concentration.

[0039] Figure 12 is a graph showing the liposome yield according to lipid content.

[0040] Figure 13 is an optical microscope (200x) image of gallnut-ethosomes according to the mixing ratio of liposomes and gallnut extracts.

[0041] Figure 14 is a graph showing the encapsulation rate of ethosomes according to the mixing ratio of liposomes and gallnut extract.

[0042] Figure 15 shows an image of a gallnut-ethosome taken with a cryo-transmission electron microscope (Cryo-TEM).

[0043] Figure 16 is a photograph showing the Franz diffusion cell permeation test method.

[0044] Figure 17 is a photograph showing a method for measuring skin moisture.

[0045] Figure 18 shows the results of skin moisture measurement.

[0046] Figure 19 shows the results of skin oil measurement.

[0047] Figure 20 is a photograph showing a method for measuring skin moisture evaporation.

[0048] Figure 21 shows the results of measuring skin moisture evaporation.

[0049] Figure 22 is a questionnaire for conducting a sensory evaluation.

[0050]

[0051] The present invention will be described in detail below according to preferred embodiments with reference to the attached drawings, but specific descriptions of configurations and operations that are readily known to those skilled in the art will be omitted. Furthermore, it should be noted that the present invention is not necessarily limited by the following embodiments, and that those skilled in the art can make various modifications to the invention within the scope of the technical concept of the invention without departing from it.

[0052] The terms used in this specification have been selected based on currently widely used general terms whenever possible, taking into account their functions in the present invention; however, these terms may vary depending on the intent of those skilled in the art, case law, the emergence of new technologies, etc. Additionally, in specific cases, terms have been arbitrarily selected by the applicant, and in such cases, their meanings will be described in detail in the relevant description of the invention. Therefore, the terms used in this invention should be defined not merely by their names, but based on their meanings and the overall content of the invention.

[0053] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as generally understood by those skilled in the art to which the present invention pertains. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and should not be interpreted in an ideal or overly formal sense unless explicitly defined in this application.

[0054] Numerical ranges include the values ​​defined in the above ranges. All maximum numerical limits given throughout this specification include all lower numerical limits as clearly written. All minimum numerical limits given throughout this specification include all higher numerical limits as clearly written. All numerical limits given throughout this specification will include all better numerical ranges within a wider numerical range, as clearly written.

[0055]

[0056] Hereinafter, an ethosome containing a gallnut extract according to a preferred embodiment of the present invention, a method for manufacturing the same, and a functional cosmetic composition using the same will be described.

[0057]

[0058] Preparation of ethosomes encapsulated with Gallnut extract

[0059]

[0060] A method for manufacturing an ethosome containing a Galla Rhois extract according to the present invention may include, as illustrated in FIG. 1, a step of manufacturing a liposome (S100); a step of manufacturing a Galla Rhois ethanol extract by extracting Galla Rhois with ethanol (S200); and a step of manufacturing an ethosome containing the Galla Rhois ethanol extract by mixing the liposome with the Galla Rhois ethanol extract (S300).

[0061]

[0062] First, the method may include a step (S100) of manufacturing a liposome. As shown in FIG. 2, this step may include: (A) a step of manufacturing a lipid film by dissolving a lipid in an organic solvent and then removing the organic solvent; (B) a step of manufacturing a liposome in an emulsion state by hydrating the lipid film with a hydration solution; and (C) a step of homogenizing the liposome.

[0063]

[0064] The step of manufacturing a lipid film by dissolving the above (A) lipid in an organic solvent and then removing the organic solvent is a step of manufacturing a lipid film by mixing and dissolving the phospholipid with an organic solvent and then completely removing the organic solvent.

[0065] As the above phospholipids, a dry powder mixed with dilinoleyl phosphatidylcholine (DLPC), palmitoyl linoleyl phosphatidylcholine (PLPC), dioleyl phosphatidylcholine (DOPC), and phosphatidylcholine (PC) may be used.

[0066] The above organic solvent may be n-hexane.

[0067] The lipid may be dissolved in the organic solvent to a concentration of 10 to 50 g / L, more preferably to a concentration of 30 to 50 g / L, and most preferably to a concentration of 40 g / L. If the lipid is added below the above range, the yield of liposomes may decrease because there is a lack of lipid molecules capable of forming a sufficient amount of liposomes. In addition, if the lipid is added above the above range, not all lipids are dissolved in the solution, resulting in a supersaturated state, which may actually decrease the yield of liposomes. That is, the supersaturated state may form irregular structures or precipitates, and lipids inside the liposomes may interact with each other to cause a phenomenon of self-digestion (self-decomposition), such as autophagy, in a biochemical context. Autodigestion in liposomes refers to the process in which lipid molecules interact with each other to break down themselves; this can reduce liposome stability and lead to inefficient liposome formation, which in turn may result in a decrease in yield.

[0068]

[0069] Subsequently, the lipid may be dissolved in an organic solvent, and then the organic solvent may be completely removed using a rotary evaporator to produce a lipid film. The lipid film produced in this manner is applied directly to the inner wall of a round-bottom flask used during the mixing of the lipid and the organic solvent and dried, and as a result, may exhibit a unique form in which the lipid layers are arranged in a double layer. At this time, if the organic solvent is not completely removed, as shown in FIG. 3(A), the hydration of the lipid film may not be properly carried out, which may cause problems in liposome formation; therefore, the process is characterized by complete drying.

[0070] For reference, FIGS. 3(B) to FIGS. 3(D) show photographs taken with a digital microscope of the double layer structure of the lipid layer when the lipid film according to one embodiment of the present invention is hydrated.

[0071]

[0072] In addition, the lipid film is characterized by not using ethanol and completely drying the volatile organic solvent. Conventionally, liposomes were prepared by dissolving lipids in ethanol and then slowly adding water to prepare ethosomes; however, due to the problem of ethanol evaporation, it was difficult to produce liposomes or ethosomes in a uniform form, such as in terms of structure and stability. Accordingly, the present invention may produce a lipid film from which organic solvents such as ethanol and hexane have been completely removed.

[0073]

[0074] The step (B) of preparing liposomes in an emulsion state by hydrating the lipid film with a hydrating solution is a step of forming liposomes in an emulsion state by hydrating the lipid layer of the lipid film with a hydrating solution (aqueous solution). In this step, other physical conditions such as temperature and concentration may affect the shape and yield of the liposomes being prepared.

[0075] The diameter of the particles of the above emulsion state liposomes may be 1 to 200 μm.

[0076]

[0077] The above hydration solution may be composed of glucose and sodium chloride (NaCl) to be similar to human body fluids, and the concentration of the hydration solution may be adjusted to have an osmotic pressure similar to the osmotic pressure range in the human body, which is 280 to 300 mOsm / kg H2O. That is, the concentration of the hydration solution may be 182 to 321 mM, more preferably 230 to 325 mM, and most preferably 265 to 293 mM.

[0078]

[0079] The lipid film may be hydrated by adding the hydration solution so that its concentration is 10 to 50 g / L, more preferably by adding the hydration solution so that its concentration is 30 to 50 g / L, and most preferably by adding the hydration solution so that its concentration is 40 g / L.

[0080] The hydration of the lipid film can be performed at a temperature of 30 to 60°C, more preferably at 50 to 60°C, and most preferably at 50°C. That is, the transition temperature (T) of the lipid m Since the transition temperature is 40°C (±5°C), at temperatures ranging from 30 to 60°C, the lipid bilayer transitions from a gel state to a liquid crystal state, possessing greater fluidity and mobility, which may promote liposome formation following the efficient hydration of the lipid film. If the temperature is below this transition temperature, the lipid molecules in the gel state become rigid and their mobility may decrease. This rigidity hinders the proper hydration of the lipid film, leading to inefficient liposome formation and low yield. In other words, the lipid molecules in the gel state have difficulty providing the flexibility necessary to spontaneously form the characteristic spherical structure of liposomes. At temperatures exceeding this transition temperature, even though the lipid bilayer remains in a liquid crystal state, excessively high temperatures can cause it to become too fluid. This extreme fluidity can disrupt the Ld. structure of the liposomes, resulting in less stable liposomes and potentially lowering the yield. Additionally, high temperatures can cause the degradation of other components involved in liposome formation, further reducing the efficiency of the process.

[0081]

[0082] Next, (C) the step of homogenizing the liposomes is a step of making the liposomes in the prepared emulsion state into a uniform shape and size. This step may be performed by homogenizing using a homogenizing device at 800 to 1,200 rpm for 40 to 80 minutes.

[0083] The diameter of the homogenized liposome particles produced through this process can be 1 to 200 μm.

[0084] At this time, since the selection of the homogenization device can affect the shape and size of the liposomes, it is most desirable to use a homomixer as the homogenization device. In fact, observation through an optical microscope revealed that using a homomixer is effective in maintaining the structural stability of liposomes compared to using a homodisper or an ultrasonic homomixer. That is, although the homodisper is mainly used for homogenizing products such as shampoo, when homogenization is performed under the same conditions, the liposomes are physically destroyed, and as a result of observation through an optical microscope (200x magnification), the shape of the liposomes cannot be found as shown in Fig. 4, which may indicate that it is not suitable for maintaining delicate structures such as liposomes. In addition, while an ultrasonic homomixer is equipment that can be used for both emulsification and mixing, it may not be suitable as homogenization equipment because when using an ultrasonic homomixer, the ultrasound aggregates the liposomes and transforms them into liposomes with a larger structure.

[0085]

[0086] Furthermore, the homogenized liposomes may further include an aging step to maintain the shape and structural stability of the liposomes. That is, the homogenized liposomes may be used after aging at 0 to 5°C for 20 to 28 hours so that the double lipid membrane of the liposomes grows sufficiently into a liquid crystal state at a low temperature.

[0087] In one embodiment of the present invention, liposomes aged at 4°C for 24 hours were observed under an optical microscope. As shown in Fig. 5, the double lipid membrane remained stable, whereas in the case of liposomes aged at a higher temperature of 23°C for 24 hours, a thinning of the membrane was observed. This may indicate that the double lipid membrane of the liposomes did not grow sufficiently into a liquid crystal state at high temperatures. These results highlight the importance of temperature control during the liposome manufacturing process and may provide useful guidelines for future research and industrial applications related to liposomes.

[0088]

[0089] Next, the method may include a step (S200) of preparing an Obeja ethanol extract (hereinafter also referred to as "Obeja Ethanol Extract" or "OEE") by extracting Galla Rhois with ethanol. This step involves grinding dried Galla Rhois into a powder and then placing it in ethanol for cold extraction.

[0090] The above-mentioned gallnut exhibits antioxidant activity through radical scavenging and redox action, and is known to be more effective than albutin in inhibiting tyrosinase activity that suppresses melanin production.

[0091] The above ethanol may be ethanol with a concentration of 60 to 80 v / v%, and it is most preferable to use ethanol with a concentration of 70 v / v%.

[0092] The above cold maceration extraction may be performed for 12 to 16 days at room temperature of 1 to 35°C with the container sealed as shown in FIG. 6 to prevent evaporation of ethanol.

[0093]

[0094] In one embodiment of the present invention, 200g of dried gallnuts may be ground into powder and then placed in 1L of 70% ethanol for cold extraction.

[0095] After the above cold extraction is completed, the extract may be centrifuged to separate the supernatant to produce an aubeja ethanol extract.

[0096]

[0097] Next, the method may include a step (S300) of mixing the liposomes with the gallnut ethanol extract to produce an etosome (hereinafter also referred to as 'gallnut-ethosome') encapsulated with the gallnut ethanol extract. This step involves producing an etosome, which is a lipid delivery system with enhanced transdermal absorption capacity, by binding ethanol to liposomes. This is illustrated in FIG. 7.

[0098] The above mixing may be performed using a vortex mixer.

[0099] The mixing ratio of the liposomes and the gallnut extract may be 4 to 7:1, and more preferably, the mixing ratio of the liposomes and the gallnut extract may be 5 to 6:1. The mixing ratio may be an important factor in determining the encapsulation rate of the ethosomes. This is because the ethanol content is determined according to the mixing ratio.

[0100] That is, the ethanol content in the ethosome may be 8 to 12%. The ethanol content may be calculated based on the mixing ratio of the liposome and the gallnut extract. For example, when the mixing ratio of the liposome and the gallnut extract is 5:1 and extraction is performed with 70% ethanol, the ethanol content is It can be calculated as.

[0101] As such, the content of the ethanol may be the most important factor in determining the encapsulation rate of the ethosomes. The ethanol is an amphiphilic small particle that, together with gallic acid in the gallnut, maintains the equilibrium of simple diffusion and osmotic pressure difference under low pH conditions, while simultaneously increasing membrane fluidity, thereby allowing the ethanol to be encapsulated instead of the hydrate inside the liposome, thus enabling the formation of the ethosome. In other words, the content of the ethanol contributes to the efficient encapsulation of the gallnut extract into liposomes, thereby enabling the formation of stable ethosomes, and thus may be the most important factor in determining the encapsulation rate of the ethosomes.

[0102]

[0103] The above mixing time may not have a major effect on determining the size or stability of the etosome. As shown in FIG. 8, even if the mixing time is shortened during the manufacturing process, it may not affect the quality of the product. Accordingly, the mixing time according to the present invention may be short, ranging from 20 to 40 seconds. Through this, productivity can be improved by minimizing the time required for the etosome manufacturing process.

[0104]

[0105] The above mixing temperature can be performed at 5 to 25°C, more preferably at 5 to 15°C, and most preferably at 10°C. When an ethosome is prepared by mixing at a temperature within this range, it may be in a stable form without cracks or deformation in the lipid layer. However, ethossomes prepared below this range may show lines such as cracks on the outside of the lipid layer. This implies that the lipid layer may become unstable at low temperatures. Ethossomes prepared above this range may exhibit deformation that appears to be stretched and expanded. This implies that the lipid layer of the ethosome may become flexible at high temperatures, causing the structure to expand. To stabilize the shape of the ethosome, which becomes unstable as the temperature rises, mixing can be performed at a temperature within this range. For reference, Figure 9 is a photograph taken at 200x magnification with an optical microscope to observe the effect of temperature on ethosome stability.

[0106]

[0107] Furthermore, in order to improve the temperature stability of the ethosome even after the above encapsulation, polyethylene glycol (PEG) 100 may be added in an amount of 5 to 10 w / w% after the ethosome is prepared. In one embodiment of the present invention, PEG 100 in an amount of 5 to 10 w / w% may be added to the encapsulated ethosome. This addition of PEG coats the surface of the ethosome, which can contribute to suppressing changes in the size and shape of the liposome despite a rise in temperature within the range of 37 to 60°C.

[0108]

[0109] Meanwhile, liposomes containing astaxanthin, an antioxidant extracted from red marine organisms, are known to be unstable under acidic conditions and stable under alkaline conditions. In contrast, the ethosome containing the gallnut extract according to the present invention is characterized by being stable under weakly acidic conditions (pH 4.5 to 5.5). This is known to be due to the characteristics of the gallnut extract, which is the substance encapsulated in the liposome. The gallnut extract contains various physiologically active components such as organic acids like gallic acid and ellagic acid, and tannins; these components may cause the gallnut extract to maintain a weakly acidic pH. Consequently, the weakly acidic pH of the gallnut ethanol extract is similar to the pH of the prepared liposome, thereby enhancing stability in ethosome formation.

[0110] In addition, the optimal pH at which the bioactive components of the above-mentioned etosome act is similar to the natural pH of the skin, so it minimizes irritation when applied to the skin, thereby enhancing skin stability and efficacy.

[0111] In one embodiment of the present invention, when an ethosome containing the gallnut extract is prepared by mixing the liposome (pH 5.13) with the gallnut ethanol extract (pH 5.24), the pH of the ethosome may be 5.19.

[0112]

[0113] The encapsulated ethosomes mentioned above may be unfiltered gallnut ethanol extract used in the step of preparing the gallnut ethanol extract. This is because high-molecular substances such as fibers and polyphenols extracted from gallnuts can be filtered out at 200 μm or less. In fact, microscopic observation of ethosomes was performed to observe changes depending on whether the gallnut extract was filtered. As shown in Fig. 10(A), the ethosomes prepared from the unfiltered gallnut ethanol extract contained fine aggregate-shaped particles inside the liposomes. That is, these particles were observed to aggregate into nano-sizes and encapsulate within the liposomes. In contrast, the ethosomes prepared from the filtered gallnut ethanol extract, as shown in Fig. 10(B), existed around the liposomes in the form of coarse granules rather than being encapsulated within the liposomes, and no encapsulation was observed. For this reason, the loss of active ingredients within the natural product gallnuts can be prevented by omitting the filtering process.

[0114]

[0115] The present invention provides an ethosome containing a gallnut extract characterized by being manufactured by the above manufacturing method.

[0116] The above ethosome may be stable under weakly acidic conditions of pH 4.5 to 5.5.

[0117]

[0118] The present invention provides a cosmetic composition using an ethosome containing the above-mentioned gallnut extract.

[0119] The above cosmetic composition may comprise an ethosome containing the above gallnut extract, aloe gel, coconut oil, olive emulsifying wax, xanthan gum, olive liquid, rose water, essential oil, grapefruit extract, and distilled water. More specifically, the above cosmetic composition may comprise, with respect to 100 parts by volume of the aloe gel, 50 to 80 parts by volume of the ethosome containing the above gallnut extract, 40 to 60 parts by volume of the coconut oil, 7 to 12 parts by volume of the above olive emulsifying wax, 0.5 to 1.5 parts by volume of the above xanthan gum, 5 to 7 parts by volume of the above olive liquid, 30 to 50 parts by volume of the above rose water, 0.5 to 1.8 parts by volume of the above essential oil, 0.2 to 0.7 parts by volume of the above grapefruit extract, and 320 to 350 parts by volume of the above distilled water. However, while the above-mentioned mixing conditions are preferable, they are not necessarily limited to the above-mentioned mixing conditions and may be appropriately adjusted as needed according to consumer needs.

[0120]

[0121] Hereinafter, the ethosome containing a Gallnut extract according to the present invention, the method for manufacturing the same, and the functional cosmetic composition using the same will be specifically described through the following examples, and the present invention is not necessarily limited only to the following examples.

[0122]

[0123] 1. Preparation of reagents

[0124] The lipids used for the preparation of the liposomes were mixed lipids in dry form, consisting of 62 wt% dilinoleyl phosphatidylcholine (DLPC), 23 wt% palmitoyl linoleyl phosphatidylcholine (PLPC), 5 wt% dioleyl phosphatidylcholine (DOPC), and 10 wt% phosphatidylcholine (PC), which were provided by Chemobis Co., Ltd. (Korea). For the preparation of the lipid film, n-hexane with a purity of 96.0% from Daejeong Chemical (Korea) was used as the organic solvent, and secondary distilled water (human power i+, Korea) was used as the aqueous phase solvent. To adjust the osmotic pressure of the aqueous phase, glucose and sodium chloride (NaCl) of Deoksan Pharmaceutical (Korea) were used as extrapure grade products, and the gallnuts to be inserted into the liposomes were domestically produced and purchased from the online sales of the agricultural corporation Duson Ae Yakcho. A rotary evaporator (EYELA N-1000, Japan), a water bath (BUCHI B-480, Switzerland), and a pH meter (STARTER 3100, Ohus, USA) were used.

[0125]

[0126] 2. Preparation of Liposomes

[0127] 2-1. Liposome yield according to osmotic concentration

[0128] 2-1-1. Experimental Method

[0129] ① Final osmolarity 182 mM (glucose concentration 56 mM)

[0130] A hydration solution was prepared by mixing a 56 mM aqueous glucose solution with a 154 mM aqueous NaCl solution (308 mOsmol / kg H2O), which serves as an electrolyte. The molar concentrations of the NaCl solution and the glucose solution were added to calculate the final molar concentration, which was then converted to an osmotic molar concentration of 182 mM. Liposomes in an emulsion state were prepared by hydrating a lipid film with the above hydration solution. (At this time, the lipid film was obtained by dissolving the mixed lipids in n-hexane to a concentration of 40 g / L and then completely removing the n-hexane using a rotary evaporator.)

[0131]

[0132] ② Final osmolarity 220 mM (glucose concentration 110 mM)

[0133] It was prepared using the same procedure as in 2-1-1.①, except that the concentration of the glucose aqueous solution was set to 110 mM. The molar concentrations of the above NaCl aqueous solution and glucose aqueous solution were added to calculate the final molar concentration, and when converted to osmotic molar concentration, the final osmotic molar concentration was 210 mM.

[0134]

[0135] ③ Final osmolarity 237 mM (glucose concentration 170 mM)

[0136] It was prepared using the same procedure as in 2-1-1.①, except that the concentration of the glucose aqueous solution was set to 170 mM. The molar concentrations of the above NaCl aqueous solution and glucose aqueous solution were added to calculate the final molar concentration, and when converted to osmotic molar concentration, the final osmotic molar concentration was 237 mM.

[0137]

[0138] ④ Final osmolarity 265 mM (glucose concentration 220 mM)

[0139] It was prepared using the same procedure as in 2-1-1.①, except that the concentration of the glucose aqueous solution was set to 2200 mM. The molar concentrations of the above NaCl aqueous solution and glucose aqueous solution were added to calculate the final molar concentration, and when converted to osmotic molar concentration, the final osmotic molar concentration was 265 mM.

[0140]

[0141] ⑤ Final osmolarity 293 mM (glucose concentration 280 mM)

[0142] It was prepared using the same procedure as in 2-1-1.①, except that the concentration of the glucose aqueous solution was set to 280 mM. The molar concentrations of the above NaCl aqueous solution and glucose aqueous solution were added to calculate the final molar concentration, and when converted to osmotic molar concentration, the final osmotic molar concentration was 293 mM.

[0143]

[0144] ⑥ Final osmolarity 321 mM (glucose concentration 340 mM)

[0145] It was prepared using the same procedure as in 2-1-1.①, except that the concentration of the glucose aqueous solution was set to 340 mM. The molar concentrations of the above NaCl aqueous solution and glucose aqueous solution were added to calculate the final molar concentration, and when converted to osmotic molar concentration, the final osmotic molar concentration was 321 mM.

[0146]

[0147] 2-1-2. Experimental Results

[0148] As described in Section 2-1-1 above, the glucose concentration was adjusted to vary the concentration of the total hydrate, and the optimal conditions for liposome production were evaluated based on the amount of liposomes generated. The liposome yield was calculated using the following formula.

[0149]

[0150]

[0151] Glucose concentration (mM) 1) Final osmolarity 2)(m M) Yield of Liposomes (%) ① 56 (1 wt%) 182 13 ② 110 (2 wt%) 210 23 ③ 170 (3 wt%) 237 28 ④ 220 (4 wt%) 265 33 ⑤ 280 (5 wt%) 293 37 ⑥ 340 (6 wt%) 321 29 1) Osmotic pressure is expressed in units of osmolality (Osmol / kg H2O), and for non-dissolving solutes, 1 molar concentration (M) is equivalent to 1 osmolality. 2) The composition of the hydrate was calculated by adding the glucose molar concentration to the electrolyte solution, which was 154 mM (0.9 wt% 308 mOsm / kg H2O), and then converting this to osmotic molar concentration.

[0152]

[0153] As a result of the evaluation, as shown in Table 1 and Figure 11 above, a large number of liposomes were formed when the glucose concentration was 280 mM. This suggests that glucose concentration is influenced by factors such as osmotic balance, solution viscosity, interactions between the solute and solvent, and the swelling process in which the lipid film absorbs water to hydrate. In other words, since glucose affects the viscosity of the solution, it may be important for the proper arrangement and stabilization of lipids during liposome formation. Additionally, the interactions between glucose, water, and sodium chloride appear to influence liposome formation. Furthermore, during the hydration of the lipid film, an appropriate glucose concentration can widen the film's spacing, facilitating better binding with the hydration solution and creating a suitable environment for liposome formation. The mixed osmotic molar concentration of glucose and sodium chloride, which determines the osmotic pressure of the solution, is shown to fall within (278+308) / 2 kg H2O=293 mM H2O, and it appears that this may actually have an effect on the formation of liposomes, which have a structure similar to living cells. Therefore, the concentration conditions of the hydration solution used in this study suggest that it is suitable for application to the human body as a cosmetic material.

[0154] These experimental results are expected to provide guidelines for the future development of liposome-based cosmetics by presenting the osmotic pressure range within which liposomes can maintain stability when applied in vivo.

[0155]

[0156] 2-2. Liposome yield according to lipid content

[0157] 2-2-1. Experimental Method

[0158] ① Lipid 1.0g → Hydrate 100mL

[0159] 1 g of the above mixed lipid was dissolved in 100 mL of n-hexane in a round-bottom flask by stirring with a magnetic stirrer. The organic solvent was completely removed using a rotary evaporator to prepare a thin lipid film on the bottom and sides of the flask. The lipid film was hydrated with 100 mL of a hydration solution made of glucose and sodium chloride (NaCl) to prepare liposomes in an emulsion state.

[0160] (At this time, the composition of the above hydrate was prepared by dissolving 9g of NaCl in 1L of water to fix the concentration at 154mM, and then using a solution with the glucose concentration adjusted to 280mM.)

[0161]

[0162] ② Lipid 2.0g → Hydrate 100mL

[0163] Except for the content of the above mixed lipid being 2.0g, it was prepared using the same procedure as 2-2-1.①.

[0164]

[0165] ③ Lipid 3.0g → Hydrate 100mL

[0166] Except for the content of the above mixed lipid being 3.0g, it was prepared using the same procedure as 2-2-1.①.

[0167]

[0168] ④ Lipid 4.0g → Hydrate 100mL

[0169] Except for the content of the above mixed lipid being 4.0g, it was prepared using the same procedure as 2-2-1.①.

[0170]

[0171] ⑤ Lipid 5.0g → Hydrate 100mL

[0172] Except for the content of the above mixed lipid being 5.0g, it was prepared using the same procedure as 2-2-1.①.

[0173]

[0174] 2-2-2. Experimental Results

[0175] The yield was calculated based on the volume of liposomes produced according to the lipid content. In this case, the liposome yield was determined by the volume of the pellet (sinking liposomes) remaining after washing the liposomes following centrifugation. More specifically, this was done by placing 1.0 mL of hydrated lipid solution into a microtube, centrifuging for 10 minutes to remove the supernatant, washing the pellet with a hydrate made of 1.0 mL glucose and NaCl, centrifuging again for 10 minutes, and measuring the volume of the pellet. Similarly, the liposome yield was calculated using the following formula, identical to that in 2-1-2. The evaluation results are shown in Table 2 and Figure 12 below.

[0176]

[0177] Amount of classified lipids (g) 1)2) Liposome yield (%) ① 1.018 ② 2.025 ③ 3.031 ④ 4.039 ⑤ 5.032* 1) Amount of dry mixed lipids added to 100ml of solution * 2) Lipid composition used: DLPC 62wt%, PLPC 23wt%, DOPC 5wt%, PC 10wt%

[0178]

[0179]

[0180] 3. Preparation of Ethosome Encapsulated with Gallnut Extract

[0181] 3-1. Ethosome encapsulation rate according to the mixing ratio of liposomes and gallnut extract

[0182] 3-1-1. Experimental Method

[0183] ① Mixing ratio (4:1)

[0184] 200g of dried gallnuts were ground into powder and placed in 1L of 70% ethanol for cold extraction at room temperature for 2 weeks. After the cold extraction was completed, the extract was centrifuged to separate the supernatant to prepare the gallnut ethanol extract. In order to investigate the effect of the volume ratio of ethanol on the encapsulation rate during the liposome encapsulation process of the gallnut ethanol extract (OEE), the liposomes obtained in 2-2-1.④ were used, and the liposomes and the gallnut ethanol extract were mixed in a ratio of 4:1 to prepare ethosomes encapsulated with the gallnut extract.

[0185]

[0186] ② Mixing ratio (5:1)

[0187] Except for the above mixing ratio being 5:1, it was manufactured using the same procedure as 3-1-1.①.

[0188]

[0189] ③ Mixing ratio (6:1)

[0190] Except for the above mixing ratio being 6:1, it was manufactured using the same procedure as 3-1-1.①.

[0191] ④ Mixing ratio (7:1)

[0192] Except for the above mixing ratio being 7:1, it was manufactured using the same procedure as 3-1-1.①.

[0193]

[0194] 3-1-2. Experimental Results

[0195] Figure 13 shows the results of optical microscopy observations showing that the encapsulation rate varied depending on the mixing ratio (4:1, 5:1, 7:1) of liposomes and Gallnut extract. To obtain a more quantitative encapsulation rate, HPLC analysis was performed using the amount of garlic acid as a labeling substance to determine the quantitative encapsulation rate. The concentration was calculated based on the peak area of ​​the garlic acid in the added OEE and the garlic acid in the Gallnut-ethosome, and the results are summarized in Table 3 below. Here, the ethanol content of the etosome was calculated based on the ratio of the added OEE. As a result, it was found that encapsulation occurred in all cases within the ethanol content range of 8–12% in the Gallnut-ethosome. This suggests that the volume ratio of ethanol has a significant influence on the liposomal encapsulation rate of OEE.

[0196]

[0197] Separation OEE mixing ratio to liposomes (ml : ml) Ethanol content (v %) 1) Gallnut-ethosome encapsulation rate (%) 2) ①4 : 1127 ②5 : 11024 ③6 : 1915 ④7 : 189 1) Ethanol content is calculated based on the mixing ratio of OEE and liposomes. Ex) In the case of a ratio of 5:1, since the volume of OEE is 1, 1 / 6 X 0.7 (extracted with 70% ethanol) = 12% 2) The encapsulation rate was calculated using the garlic acid HPLC analysis value as the ratio of the encapsulated OEE to the added OEE.

[0198]

[0199] 3-2. Cryo-TEM Analysis

[0200] As a result of the experiment '3-1. Ethosome encapsulation rate according to the mixing ratio of liposomes and gallnut extract' above, cryo-transmission electron microscopy (Cryo-TEM) analysis was performed on the sample that showed the highest gallnut-ethosome encapsulation rate (%) (② OEE mixing ratio to liposomes = 5:1).

[0201] Figure 15 is an image of the gallnut-ethosome taken using cryo-transmission electron microscopy (Cryo-TEM). Observation results clearly identified spherical particles with an average diameter of approximately 200 nm, indicating that the ethosome was formed normally. Each particle exhibited a distinct phospholipid bilayer structure, and a dense region with high electron density existed inside. This high electron density region within suggests that polyphenolic components such as gallic acid and ellagic acid are encapsulated within the gallnut extract.

[0202] In addition, a multivesicular structure containing multiple intraluminal vesicles was observed in some particles. This multivesicular structure has an additional lipid membrane formed inside the outer membrane, which enables sustained release of the contents and can contribute to improved persistence within the skin and transdermal absorption efficiency.

[0203] As such, through cryo-TEM analysis, it was confirmed that the gallnut-ethosome according to the present invention has a complex nanostructure (multilayer-multivesicular hybrid) rather than a simple single-membrane liposome structure. This is believed to have been formed by the increased fluidity of the phospholipid membrane in the presence of ethanol and the gallnut extract inducing membrane rearrangement. Consequently, the gallnut ethosome of the present invention is structurally stable and demonstrates that it possesses high encapsulation efficiency and continuous release characteristics simultaneously.

[0204]

[0205] 3-3. Skin penetration experiment of ethosomes encapsulated with Gallnut extract

[0206] In order to determine whether ethosomes containing gallnuts prepared under optimal conditions penetrate the actual skin well, and how much OEE, a simple gallnut extract not encapsulated in ethosomes, penetrates the skin, we conducted a study in collaboration with the Skin Clinical Support Center of the Jeju Industry-Academic Convergence Institute based on the in vitro skin absorption test guidelines announced by the Ministry of Food and Drug Safety.

[0207]

[0208] 3-3-1. Skin Absorption Test Method

[0209] The transdermal absorption system used in the skin absorption experiment was the Franz diffusion cell system DHC-6TD, SYSTEM 918-6 (LOGAN Instruments, USA). PB-M (Permeation Barrier Membrane, LOGAN Instruments, USA) was used as the artificial skin. PIC (EDTA-free Protease Inhibitor Cocktail, Roche, Switzerland), a protease inhibitor, was used as a reagent for pretreatment of the eluted samples obtained from the Franz diffusion cell; RIPA buffer (BIOSESANG, Korea) was used for cell membrane degradation; and DPBS (Dulbecco's Phosphate Buffered Saline Liquid, BIOSESANG, Korea) was used as the cell culture medium. Gallic acid (Sigma-Aldrich, USA) and methyl gallate (Sigma-Aldrich, USA) were used as standard reagents for HPLC analysis, and HPLC-grade formic acid (Sigma-Aldrich, USA), distilled water (JTBaker, USA), acetonitrile (Fisher, USA), and MeOH (Fisher, USA) were used as mobile phase solvents.

[0210] Franz diffusion cell permeation tests were performed in the manner shown in Fig. 16. After applying 0.5 mL each of OEE and Gallnut ethosomes to artificial skin, the HPLC chromatogram area values ​​of samples collected at 1, 2, 4, 7, 10, and 24 hours were substituted into the calculated regression line equation to represent the permeation amount of gallic acid over time. For the HPLC analysis, a Kromasil C18 column (4.6 × 250 mm, 5 μm, Sweden) was used, and the HPLC system consisted of an Alliance e2695 XE separations module and a 2998 photodiode array detector (PDA, Waters, USA). The detector wavelength was 253 nm, and gradient elution was performed using a 0.1% formic acid and acetonitrile solution. Waters' Empower system was used for data analysis.

[0211] The eluted sample obtained from the Franz diffusion cell was pretreated by mixing RIPA buffer : 10X PIC : DPBS = 1 : 1 : 8 and reacting it in an ice bucket for 1 hour before use in analysis. The standard solution was prepared to a concentration of 1000 ppm by mixing the gallic acid and methyl gallite standard with MeOH. A calibration curve was constructed by serially diluting the prepared standard solution to different concentrations. Subsequently, the content of gallic acid and methyl gallite was determined from the prepared test solution using a regression line equation calculated from the chromatogram area values. Since there was a difference in methyl gallite content before and after filtering, gallic acid was used as the reference.

[0212]

[0213] 3-3-2. Skin Absorption Test Results

[0214] The eluted samples obtained from the Franz diffusion cell were analyzed using High Performance Liquid Chromatography (HPLC). During this process, gallic acid was set as the labeling agent. The results showed that the *Galleria mellonella* ethosomes exhibited high permeability in the initial stages, indicating rapid absorption into the artificial skin. Specifically, the total amount of gallic acid from the *Galleria mellonella* ethosomes permeated through the Franz diffusion cell over 24 hours was measured at 50 ppm, corresponding to a total permeability of 43%. In contrast, the total amount of gallic acid from the *Galleria mellonella* ethanol extract (OEE) eluted from the Franz diffusion cell during the same period was 22 ppm, with a total permeability of only 19%. This indicates that the *Galleria mellonella* ethosomes showed approximately twice the levels of overall permeability and permeability compared to OEE. Furthermore, the *Galleria mellonella* ethosomes exhibited a stable state, maintaining a permeability of 10–20% continuously from the beginning for 24 hours. In contrast, OEE exhibited an uneven pattern, with most penetration occurring only after 12 hours. This implies that while gallnut-ethosome is absorbed into the skin more quickly and steadily, OEE's absorption begins only after a certain amount of time. These results suggest that when OEE is applied as a cosmetic, the active ingredient may only penetrate the skin after 12 hours, making it difficult to expect the skin effects of the active ingredient as it may be washed away before absorption.

[0215] Therefore, it is considered that applying cosmetics in the form of gallnut-ethosome is more effective. Gallnut-ethosome has the advantage of being rapidly absorbed into the skin to continuously supply active ingredients, so it will demonstrate superior efficacy during actual cosmetic development.

[0216]

[0217] 4. Cosmetic composition using an ethosome encapsulated with gallnut extract

[0218] 4-1. Preparation of cosmetic compositions

[0219] (Comparative Example 1)

[0220] First, to mix the oil phase, gallnut ethosome, coconut oil, and olive emulsifying wax were placed in a beaker, and this mixture was stirred using a homogenizer at 70°C at a speed of 3,000 rpm. Next, to prepare the aqueous phase, aloe gel, rose water, grapefruit extract, and distilled water were stirred at 70°C at a speed of 3,000 rpm for 5 minutes. The temperature-adjusted aqueous phase was gradually added to the stirring oil phase. After further stirring the integrated mixture of the aqueous and oil phases for 30 minutes, grapefruit extract and essential oil were added to prepare Comparative Example 1 (hereinafter also referred to as 'BASE').

[0221]

[0222] (Comparative Example 2)

[0223] The composition was prepared in the same manner as Comparative Example 1, except that an ethanol extract of *OEE* was added during the preparation of the aqueous phase. Through this, the cosmetic composition of Comparative Example 2 was prepared. (Hereinafter referred to as 'BASE+OEE'.)

[0224]

[0225] (Example 1)

[0226] The cosmetic composition of Example 1 was prepared in the same manner as Comparative Example 1, except that a Gallnut ethosome was added during the process of preparing the aqueous layer. Through this, the cosmetic composition of Example 1 was prepared. (Hereinafter also referred to as 'BASE+OEE+ETOSOME'.)

[0227]

[0228] The raw materials introduced into the reactor to manufacture Comparative Examples 1 and 2 and Example 1 are as shown in [Table 4] below.

[0229]

[0230] (Unit: parts by volume) Composition Comparative Example 1 Comparative Example 2 Example 1 Gallnut Ethanol Extract--60 Gallnut Ethosome-60 Aloe Gel 100 100 100 Coconut Oil 50 50 50 Olive Emulsifying Wax 10 10 10 Xanthan Gum 11 1 Olive Liquid 5.5 5.5 5.5 Rose Water 40 40 40 Essential Oils (Geranium + Lavender + Rosewood) 1.0 1.0 1.0 Grapefruit Extract 0.5 0.5 0.5 Distilled Water 3 3 2 3 3 2 3 3 2

[0231]

[0232] 4-2. Evaluation of Cosmetic Compositions

[0233] 4-2-1. Skin Moisture Measurement Method and Results

[0234] Skin moisture is measured using capacitance at a depth of 30–40 μm in the stratum corneum with a diameter of 16 nm. Since the skin's electrostatic load and moisture retention are proportional, a low value indicates dryness, while a high value indicates a high moisture content. The measurement unit is expressed in the arbitrary unit (AU). According to recent studies, for the face, values ​​below 30 AU were interpreted as very dry, 30 to less than 50 as dry, 50 to less than 60 as moderate, and 60 or higher as moist. For the arms, hands, legs, and elbows, values ​​below 15 AU were considered very dry, 15 to less than 29 as dry, 30 to less than 39 as moderate, and 40 or higher as moist.

[0235] In this test, changes in arm moisture were measured in the manner shown in Fig. 17. The device used for measuring skin moisture was the Corneameter® (CM825, Courage+Khazaka, Germany), which measures skin moisture by measuring the electrical capacitance of the conducted current using the electrode spacing in contact with the skin surface. During measurement, excessive force should not be applied, and care must be taken to ensure that only one end does not make contact. During each measurement, care was taken to ensure that the probe made consistent and flat contact with the skin.

[0236]

[0237] The evaluation results are shown in Figure 18. The subjects' baseline moisture content ranged from 30 to 50, with an average of 42. The skin of most subjects was in an above-average moist state. Thirty minutes after applying the cosmetic, the moisture levels measured averaged 63 for BASE, 57 for BASE+OEE, and 67 for BASE+OEE+ETOSOME. The lower moisture content of BASE+OEE compared to BASE is thought to be due to moisture evaporation caused by the presence of ethanol in OEE. Despite the presence of ethanol, BASE+OEE+ETOSOME was confirmed to have the highest moisturizing capacity among the three test groups. Sixty minutes after applying the cosmetic, the moisture levels measured averaged 58 for BASE, 61 for BASE+OEE, and 67 for BASE+OEE+ETOSOME. Unlike the moisture content after 30 minutes, the fact that the moisture content of BASE+OEE became higher than that of BASE at 60 minutes suggests that the gallnut enhances skin moisturizing power after the ethanol in OEE evaporates. It was confirmed that BASE+OEE+ETOSOME maintained the same moisturizing power as 30 minutes, showing almost no change in moisture content.

[0238]

[0239] 4-2-2. Skin Oil Measurement Method and Results

[0240] Skin oil content is measured by assessing the level of light transmission (photometric reflection principle); generally, for dry skin, the oil content of the arms, hands, legs, and elbows is 0–6 μg / cm² 2 , for neutral, 6 μg / cm² 2The following is not measured in the case of oily skin. Meanwhile, skin oil content was measured using the Sebumeter® SM815 (Courage + Khazaka electronic GmbH, Germany). Sebum was absorbed by pressing the probe, which has a translucent lipid-absorbing tape attached to the measurement area, onto the skin surface to be measured for 10 seconds using the sebumeter cassette, and then the oil content was measured using the measuring device.

[0241]

[0242] The evaluation results are shown in Figure 19. In this test, the initial oil content of most subjects was 0, except for 25%, as the oil content of the arms after 30 minutes was unclear. As a result of measuring the oil content after 1 hour, the average was 16 for BASE, 34 for BASE+OEE, and 37 for BASE+OEE+ETOSOME. The oil content of BASE+OEE and BASE+OEE+ETOSOME was more than double that of BASE, indicating that even the gallnut extract without etosome treatment increases the oil content of the skin.

[0243]

[0244] 4-2-3. Method for Measuring Skin Moisture Evaporation

[0245] To measure skin barrier function, transepidermal water loss (TEWL) was evaluated using the Tewameter™ Hex (Courage + Khazaka electronic GmbH, Germany), as shown in Fig. 20. For the measurement of skin water evaporation, 30 sensors located on each of the six sides of the instrument probe, acting like cameras, check relative humidity and temperature; based on a large amount of data regarding the skin surface and the instrument probe environment, the results are expressed as transepidermal water loss (g / h / m²). 2It is indicated as ). If the skin barrier is damaged, loss increases and the result value is high, while the stronger the barrier, the significantly lower the water evaporation, with transepidermal water loss being 0–9 g / h / m². 2 It was defined that 10–14 is a very healthy state, 15–24 is a healthy state, 25–29 is a worrying state, and 30 or more is a very poor state.

[0246]

[0247] The evaluation results are shown in Figure 21. The average moisture evaporation rate 30 minutes after cleansing was 12, indicating that all subjects possessed a healthy skin barrier. When the skin moisture evaporation rate was measured 30 minutes after applying the cosmetic product, the results were an average of 11 for BASE, 11 for BASE+OEE, and 10 for BASE+OEE+ETOSOME. The moisture evaporation rates for BASE and BASE+OEE showed similar values, which is thought to be the reason why moisture evaporation decreased after applying the cosmetic product compared to before. Notably, the initial value of 12 (without any product applied) decreased to 10 after 30 minutes and 9 after 60 minutes for BASE+OEE+ETOSOME. This suggests that the ethosome-treated Gallnut extract penetrates the skin barrier, strengthening the skin barrier function from a healthy state to a very healthy state.

[0248]

[0249] 4-2-4. Sensory Evaluation Methods and Results

[0250] A sensory evaluation was conducted using a questionnaire as shown in Fig. 22. To analyze user satisfaction after using the gallnut etosome, five questions were prepared, and the answers were given on a scale of 5: very good (4), good (3), average (2), bad (1), and very bad (0). For the items such as skin moisturization in the survey, the mean value, standard deviation, and the percentage of the number of subjects for the answers were calculated.

[0251] Regarding the survey results on the user experience of the etosome cosmetic of the present invention, 100% of the subjects rated all evaluation items as average or higher. Regarding the best aspects after use, 87.5% of subjects indicated that their skin felt smooth, and 75% indicated that it felt moisturizing. There were no reports of any specific adverse skin reactions during the testing of the Gallnut Ethosome Serum. Although one subject reported feeling irritated when applying BASE+OEE+ETOSOME, the irritation was not severe enough to stop the test. A detailed consultation revealed that the subject had sensitive skin due to atopic dermatitis. This indicates that while skin with wounds may feel irritated due to the influence of ethanol, healthy skin showed high satisfaction regarding the smooth and moisturizing effects after use.

[0252]

[0253] Although the ethosome containing a gallnut extract according to the preferred embodiment of the present invention, the method for manufacturing the same, and the functional cosmetic composition using the same have been described as described above, this is merely an example and those skilled in the art will understand that various changes and modifications are possible within the scope of the technical concept of the present invention.

Claims

1. Step of manufacturing a liposome (S100); Step (S200) of preparing an ethanol extract of Galla Rhois by extracting Galla Rhois with ethanol; and A method for preparing an ethosome encapsulated with a gallnut extract, characterized by including the step (S300) of mixing the liposomes with the gallnut ethanol extract to prepare an ethosome encapsulated with the gallnut ethanol extract.

2. In Paragraph 1, The step (S100) of manufacturing the above liposome is, (A) A step of dissolving lipids in an organic solvent and then removing the organic solvent to produce a lipid film; (B) a step of preparing liposomes in an emulsion state by hydrating the lipid film with a hydration solution; and (C) A method for preparing an ethosome encapsulated with gallnut extract, characterized by including the step of homogenizing the liposomes.

3. In Paragraph 2, In the step of manufacturing a lipid film by dissolving the above (A) lipid in an organic solvent and then removing the organic solvent, A method for preparing an ethosome encapsulated with a Gallnut extract, characterized by dissolving the lipid in the organic solvent such that the concentration of the lipid is 10 to 50 g / L.

4. In Paragraph 2, In the step of (B) hydrating the lipid film with a hydration solution to produce a liposome in an emulsion state, A method for preparing an ethosome encapsulated with a Gallnut extract, characterized in that the concentration of the above-mentioned solution is 230 to 325 mM.

5. In Paragraph 1, In the step (S300) of manufacturing the above ethosome, A method for preparing an ethosome encapsulated with gallnut extract, characterized in that the mixing ratio of the liposome and the gallnut extract is 4 to 7:

1.

6. Ethosome containing a Gallnut extract characterized by being manufactured by the manufacturing method of Claim 1 7. In Paragraph 6, The above ethosome is, Ethosome encapsulated with a Gallnut extract characterized by being stable under weakly acidic conditions of pH 4.5 to 5.5 8. A cosmetic composition characterized by comprising an ethosome encapsulated with a Gallnut extract according to claim 6.

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