Method for increasing polyphenol active ingredients using high-temperature and pressurized steam extraction

Abstract

This invention provides an extraction method that can effectively increase the extraction yield of polyphenols and flavonoids. [Solution] A method for producing an extract with increased polyphenol and flavonoid content, comprising the steps of: a) washing and drying the raw material and then grinding it to a size of 0.3 to 0.7 mm; b) extracting the extract by adding high-temperature steam under a pressure of 8 to 10 bar; and c) filtering the extract and then concentrating it. The raw material may be one selected from the group consisting of camellia, evergreen oak, Japanese cinnamon, and edible mushrooms.

Description

[Technical Field]

[0001] This invention relates to an extraction method that can increase the amount of useful components such as polyphenols and flavonoids by utilizing a high-temperature, pressurized steam extraction process. [Background technology]

[0002] The skin, while directly exposed to the external environment, has the function of protecting itself from temperature, humidity, antigens, and ultraviolet rays. However, physical and chemical stimuli such as external pollutants, ultraviolet rays, and stress can impair skin function. As we age, cell proliferation and immune activity weaken, making it difficult to quickly repair skin cell damage. This leads to an overall deterioration of skin components, resulting in wrinkles, loss of elasticity, and keratinization.

[0003] Skin aging can be broadly divided into intrinsic aging and extrinsic aging. Intrinsic aging is a naturally occurring aging phenomenon, clinically characterized by a decrease in elasticity, rough skin texture, deep wrinkles, and pigmentation. Extrinsic aging is an aging phenomenon caused by external environmental factors such as ultraviolet rays, reactive oxygen species, and stress. Recently, rapid industrialization and the resulting air pollution and ozone layer depletion have increased UV levels, which generate powerful oxidizing substances such as reactive oxygen species. These UV rays induce the destruction of connective tissue formation such as skin collagen, inhibition of cell membrane function, promotion of DNA mutation, deformation of protein function, and deformation of intracellular signaling molecules, leading to wrinkle formation, pigmentation, and other skin aging effects. To prevent such skin damage, cosmetics and skin protectants containing a variety of skin-protective ingredients such as retinol, retinoids, vitamin C, flavonoids, tocopherol, and coenzyme Q10 have been developed.

[0004] In particular, ultraviolet rays (UV) from sunlight are a well-known cause of aging. Prolonged exposure to UV rays thickens the stratum corneum, and the collagen and elastin, the main components of skin, denature, leading to a loss of skin elasticity. While collagen and elastin are regulated by various factors, the expression of metalloproteinases such as collagenase and elastase leads to the breakdown of collagen and elastin, resulting in a decrease in the collagen content within the skin. Various substances have been developed and used to suppress this decrease in collagen and elastin, which causes elasticity loss. Among these, retinol and retinoic acid show elasticity-improving effects, and protein fractions obtained from leguminous seeds have also been applied to increase elasticity. However, these retinoids have the drawback of being sensitive to skin irritation and allergies even when applied to the skin in very small amounts.

[0005] Recently, in order to develop cosmetics that are relatively less toxic and gentle on the environment, we have been using extraction processes and fermentation technologies that utilize natural products to search for bioactive substances with properties such as whitening, anti-aging, anti-wrinkle, antioxidant, and anti-inflammatory effects, and using these as cosmetic ingredients.

[0006] In this invention, we sought an extraction method that could extract large quantities of useful components such as polyphenols and flavonoids. We conducted experiments by crushing natural raw materials into powder and applying a high-temperature and high-pressure extraction process to the powdered raw materials. As a result, we confirmed that the extract obtained by the high-temperature and high-pressure process had a significantly increased content of polyphenols and flavonoids compared to the extraction methods known from the past, thus completing the present invention. [Prior art documents] [Patent Documents]

[0007] Korean Published Patent No. 10-2017-0122684 [Overview of the project] [Problems that the invention aims to solve]

[0008] Therefore, an object of the present invention is to provide an extraction method that can effectively increase the extraction yield of polyphenols and flavonoids.

[0009] Another object of the present invention is to provide an extract having an increased content of polyphenols and flavonoids produced by the extraction method described above.

[0010] Another object of the present invention is to provide a cosmetic composition containing the extract as an active ingredient. [Means for solving the problem]

[0011] In order to achieve the above-mentioned objectives of the present invention,

[0012] The present invention provides a method for producing an extract with increased polyphenol and flavonoid content, comprising the steps of: a) washing and drying the raw materials and then grinding them to a size of 0.3 to 0.7 mm; b) extracting the extract by adding high-temperature steam under a pressure of 8 to 10 bar; and c) filtering the extract and then concentrating it.

[0013] In one embodiment of the present invention, the raw material may be one selected from the group consisting of camellia, evergreen oak, Japanese cinnamon, and edible mushrooms.

[0014] In one embodiment of the present invention, the polyphenol may be one or more selected from the group consisting of 4-hydroxybenzoic acid, coumaric acid, rutin, naringenin, gallic acid, protocatechuic acid, chlorogenic acid, catechin, and tannic acid.

[0015] In addition, the present invention provides an extract with increased contents of polyphenols and flavonoids produced by a method.

[0016] In addition, the present invention provides a cosmetic composition containing the extract as an active ingredient.

Effects of the Invention

[0017] The high-temperature and pressurized steam extraction step of the present invention can significantly increase the extraction contents of protocatechuic acid and chlorogenic acid, etc., together with gallic acid which is a powerful antioxidant as a useful ingredient. In addition, the extraction contents of catechin and tannic acid, which are effective in suppressing wrinkle formation, can also be effectively increased. The extract extracted by such an extraction method contains polyphenol and flavonoid components in high contents. Therefore, the composition of the present invention containing the extract as an active ingredient can be usefully used as various functional cosmetic compositions such as antioxidant, wrinkle-improving and whitening compositions.

Brief Description of the Drawings

[0018] [Figure 1] FIG. 1 is an extraction process diagram briefly showing the high-temperature and pressurized steam extraction step of the present invention.

Modes for Carrying Out the Invention

[0019] The present invention is characterized in that it provides a method for producing an extract with increased contents of polyphenols and flavonoids, including: a) a step of washing and drying a raw material and then pulverizing it to a size of 0.3 to 0.7 mm; b) a step of adding high-temperature steam under a pressure of 8 to 10 bar to extract an extract; and c) a step of filtering and then concentrating the extract.

[0020] In the step a), the raw material is various medicinal or edible plants that can be a target for extraction.

[0021] In one specific example of the present invention, the raw material may be camellia, evergreen oak, Japanese cinnamon, or edible mushroom, but is not particularly limited to that type.

[0022] The "Camellia japonica L." referred to herein is an evergreen broad-leaved small tree belonging to the family Tea family, growing in Gochang, North Jeolla Province; Haenam, Wando, Gangjin, Yeosu, Gwangyang, Geoje, South Gyeongsang Province; the southern coastal region; and Jeju Island. The tree grows to a height of approximately 15m and a diameter of approximately 50cm. The upper surface of the leaves is dark green and glossy, while the underside is yellowish-green. The leaves are oval-shaped, arranged oppositely, 5-12cm long and 3-7cm wide, with fine, wavy serrations.

[0023] The "Akagashi (Quercus acuta Thunb.)" referred to herein belongs to the beech family and is also known as "Casenum, Karannip (Jeolla Province), Kasingu, Bukkasingu, Hongkasingu, and Polenang (Jeju Island)." It is distributed in sunny mountain foothills and valleys, reaching a height of 20m and a diameter of 60cm. The trunk grows straight, and with countless branches and lush leaves, it forms a magnificent tree shape. As a temperate evergreen broad-leaved tree, it is a common species in the temperate regions of Korea and Japan. Akagashi is the main tree that forms the temperate forests of the southern islands, including Jeju Island, and the southern coastal mountains. One theory is that the name Akagashi comes from the fact that the wood has a reddish tint. The fruit of Akagashi contains tannins that have an astringent effect and stop diarrhea, and is known to have medicinal effects on dysentery, colitis, diarrhea, breast pain, gingivitis, and stomatitis.

[0024] The "Cinnamomum japonicum Siebold" referred to herein is an evergreen, broad-leaved tree belonging to the laurel family, with a pungent and sweet taste. In traditional Chinese medicine, the bark and fruit are used as a medicine called "Tianzhu Gui," and these bark and fruit contain components such as phellandrene, eugenol, and methyleugenol, which are reported to be effective in improving digestion and treating vomiting, dysentery, and neuralgia. In addition, the leaves of the Japanese cinnamon tree have a strong aroma and have been used as a tea substitute since ancient times.

[0025] To date, hot water extraction and organic solvent extraction have been the main methods used to extract useful components from the plants mentioned above.

[0026] In this invention, the polyphenol and flavonoid content of extracts from camellia, evergreen oak, and Japanese cinnamon was measured using different extraction methods. As a result, it was confirmed for the first time that the polyphenol and flavonoid content was significantly increased in the extracts obtained using the high-temperature, pressurized steam extraction method of this invention compared to conventionally known extraction methods.

[0027] The aforementioned polyphenols include 4-hydroxybenzoic acid, coumaric acid, rutin, naringenin, gallic acid, protocatechuic acid, chlorogenic acid, catechin, and tannic acid.

[0028] Furthermore, the present invention provides an extract having an increased content of polyphenols and flavonoids produced by the above method.

[0029] The aforementioned extract contains large amounts of gallic acid, a powerful antioxidant, along with protocatechuic acid and chlorogenic acid as beneficial components, and also contains large amounts of catechin and tannic acid, which are effective in suppressing wrinkle formation.

[0030] Furthermore, the present invention provides a cosmetic composition containing the extract as an active ingredient.

[0031] The cosmetic composition of the present invention can be used for a variety of functional purposes, such as antioxidant, wrinkle improvement, and skin whitening.

[0032] In one specific example of the present invention, the extract may be included in the cosmetic composition at a concentration of 0.0001 to 2000 μg / ml.

[0033] Products to which the cosmetic composition of the present invention can be added include, for example, cosmetics such as astringent lotions, softening lotions, nourishing lotions, various creams, essences, masks, and foundations, as well as cleansers, facial washes, soaps, treatments, and serums.

[0034] Specific dosage forms of the cosmetic composition of the present invention include one dosage form selected from the group consisting of softening lotion, gel, water-soluble liquid, milk lotion, nourishing cream, massage cream, essence, oil-in-water emulsion, water-in-oil emulsion, paste anhydrous product, solid anhydrous product, oil dispersion in an aqueous phase using microspheres, ionic lipid vesicles, nonionic lipid vesicles, ointment, cleansing foam, cleansing water, pack, body oil, oil-in-water makeup base, water-in-oil makeup base, foundation, skin cover, lipstick, lip gloss, face powder, two-way cake, eyeshadow, mascara, cheek color, and eyebrow pencils.

[0035] According to a preferred embodiment of the present invention, the content of the active ingredient of the present invention (extract extracted by high-temperature, pressurized steam extraction) is 0.00001 to 40% by weight, preferably 0.0005 to 40%, and more preferably 0.0005 to 20% by weight, relative to the total weight of the composition.

[0036] On the other hand, the cosmetic composition according to the present invention can also be stabilized and formulated by incorporating the active ingredient (extract extracted by high-temperature, pressurized steam extraction) inside nanoliposomes. By incorporating the active ingredient inside nanoliposomes, the active ingredient is stabilized, solving problems such as precipitate formation, discoloration, and odor change during formulation, increasing the solubility and transdermal absorption rate of the ingredient, and maximizing the expected efficacy from the extract.

[0037] In this invention, nanoliposomes are defined as liposomes having the morphology of conventional liposomes, with an average particle size of 10 to 500 nm. In preferred embodiments of this invention, the average particle size of nanoliposomes is 50 to 300 nm, and more preferably 100 to 200 nm. When the average particle size of nanoliposomes exceeds 500 nm, the improvement in skin penetration and the improvement in dosage form stability, which are among the technical effects to be achieved in this invention, are very weak.

[0038] The nanoliposomes used to stabilize the active ingredient (extract extracted by high-temperature, pressurized steam extraction) according to the present invention can be produced from a mixture containing a polyol, an oily component, a surfactant, a phospholipid, a fatty acid, and water.

[0039] The polyol used in the nanoliposomes of the present invention is not particularly limited, and is preferably one or more selected from the group consisting of propylene glycol, dipropylene glycol, 1,3-butylene glycol, glycerin, methylpropanediol, isopropylene glycol, pentylene glycol, erythritol, xylitol, sorbitol, and mixtures thereof. The amount used is 10 to 80% by weight, preferably 30 to 70% by weight, based on the total weight of the nanoliposomes.

[0040] The oily component used in the production of nanoliposomes of the present invention can utilize a variety of oils known in the industry, preferably hydrocarbon oils such as hexadecane and paraffin oil, ester-based synthetic oils, silicone oils such as dimethicone and cyclomethicone, animal and vegetable oils such as sunflower oil, corn oil, soybean oil, avocado oil, sesame oil and fish oil, ethoxylated alkyl ether oils, propoxylated alkyl ether oils, sphingonoid lipids such as phytosphingosine, sphingosine and sphinganin, cerebroside cholesterol, sitosterol cholesterol sulfate, sitosterol sulfate, C 10‐40 This refers to fatty alcohols and mixtures thereof. The amount used may be 1.0 to 30.0% by weight relative to the total weight of the nanoliposomes, preferably 3.0 to 20.0% by weight.

[0041] Any surfactant known in the art can be used in the production of nanoliposomes according to the present invention. For example, anionic surfactants, cationic surfactants, amphoteric surfactants, and nonionic surfactants can be used. Anionic surfactants and nonionic surfactants are preferred. Specific examples of anionic surfactants include alkyl acyl glutamates, alkyl phosphates, alkyl lactylates, dialkyl phosphates, and trialkyl phosphates. Specific examples of nonionic surfactants include alkoxylated alkyl ethers, alkoxylated alkyl esters, alkyl polyglycosides, polyglyceryl esters, and sugar esters. Particularly preferred surfactants are polysorbates belonging to the nonionic surfactant category. The amount used may be 0.1 to 10% by weight of the total weight of the nanoliposomes, and preferably 0.5 to 5.0% by weight.

[0042] The phospholipids used as further components in the production of the nanoliposomes of the present invention are amphiphilic lipids and include natural phospholipids (e.g., egg yolk lecithin or soy lecithin, sphingomyelin) and synthetic phospholipids (e.g., dipalmitoylphosphatidylcholine or hydrogenated lecithin), preferably lecithin. In particular, naturally derived unsaturated or saturated lecithin extracted from soybeans or egg yolks is preferred. Typically, naturally derived lecithin contains 23-95% phosphatidylcholine and 20% or less phosphatidylethanolamine. In the production of the nanoliposomes of the present invention, the amount of phospholipids used is 0.5-20.0% by weight of the total weight of the nanoliposomes, preferably 2.0-8.0% by weight.

[0043] The fatty acids used in the nanoliposome production of the present invention are preferably higher fatty acids, and C 12‐22 Examples of saturated or unsaturated fatty acids in the alkyl chain include lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, and linoleic acid. The amount used may be 0.05 to 3.0% by weight relative to the total weight of the nanoliposomes, preferably 0.1 to 1.0% by weight.

[0044] The water used in the production of nanoliposomes according to the present invention is generally deionized distilled water, and the amount used may be 5.0 to 40% by weight relative to the total weight of the nanoliposomes.

[0045] Nanoliposomes can be manufactured by various methods known to the industry, but most preferably by applying a mixture containing the aforementioned components to a high-pressure homogenizer. Nanoliposome manufacturing by high-pressure homogenizer can be carried out under various conditions (e.g., pressure, number of passes, etc.) depending on the desired particle size, and preferably nanoliposomes can be manufactured by passing the mixture through the high-pressure homogenizer 1 to 5 times under a pressure of 600 to 1200 bar.

[0046] The cosmetic composition of the present invention can be used alone or in combination with other cosmetic compositions, or in combination with other cosmetic compositions not of the present invention. Furthermore, the cosmetic composition of the present invention can be used according to the usual method of use, and the number of applications can be varied according to the user's skin condition or preference.

[0047] The present invention will be described in more detail below with reference to examples. These examples are provided to illustrate the present invention more concretely, and the scope of the present invention is not limited to these examples.

[0048] <Example 1>

[0049] Production of Camellia, Japanese evergreen oak, and Japanese cinnamon extracts by high-temperature and high-pressure extraction process

[0050] The camellia leaves used in this experiment were collected from the Wando Arboretum, the branches of Quercus acuta Thunb. were collected from the Jeollanam-do Forest Resources Research Institute, and the leaves and branches of Cinnamomum japonicum Siebold were also collected from the Jeollanam-do Forest Resources Research Institute.

[0051] Each of the natural raw materials prepared above was washed and dried, then ground to a size of 0.3-0.7 mm and used as a powder. Each ground natural raw material was compressed and loaded into high-temperature and high-pressure equipment, and extracted in water at 150-200°C for 5 minutes. The extracted extract was filtered through a 0.7 μm size filter to remove microorganisms and foreign matter. The filtered filtrate was concentrated using a rotary vacuum evaporator to a concentration of 50-1,000 μg / mL and used. The manufacturing process of camellia, red oak, and Japanese cinnamon extracts using the high-temperature and high-pressure extraction process of the present invention is shown in detail in Figure 1.

[0052] The extract yield was calculated using the following formula 1.

[0053] [Formula 1]

[0054] Yield = (Weight of sample after drying (g) / Weight of sample before extraction (g)) × 100

[0055] [Table 1]

[0056] <Comparative Example 1>

[0057] Production of Camellia, Japanese evergreen oak, and Japanese cinnamon extracts using existing extraction processes.

[0058] Each of the prepared natural raw materials was washed and dried, then ground to a size of 0.3-0.7 mm to be used as a powder. 100 g of leaf powder was immersed in 1,000 mL of distilled water, and then extracted using an autoclave at 100°C for 10 minutes. The extracted extract was filtered through filter paper (Whatman No. 2), concentrated using a rotary vacuum evaporator, and used as a sample.

[0059] The extraction yield was calculated using Equation 1 above.

[0060] [Table 2]

[0061] <Experimental Example 1>

[0062] Evaluation of the antioxidant activity of the extract

[0063] To evaluate the antioxidant activity of the camellia, evergreen oak, and cinnamon extracts prepared in Example 1 and Comparative Example 1, the DPPH free radical scavenging activity and ABTS radical scavenging activity were investigated.

[0064] <1-1>DPPH free radical scavenging ability

[0065] DPPH free radical testing is a widely used method for measuring the antioxidant activity of natural extracts. In this experiment, the radical scavenging ability of samples was measured by modifying Blois's method, one of the methods for measuring antioxidant activity. Specifically, 800 μL of 0.25 mM DPPH (2,2-diphenyl-1-picrylhydrazyl, Sigma, USA) dissolved in methanol and 200 μL of each sample (50-1,000 μg / mL) were placed in EP tubes and reacted in a dark room for 15 minutes by vortexing. Subsequently, absorbance was measured at 517 nm using an HT multi-detection microplate reader. The scavenging ability (IC) of the extracts against DPPH was measured. 50 The concentration () is expressed as the concentration required to reduce the absorbance of the control group using only the solvent by 50%.

[0066] TIFF2026079819000004.tif17165

[0067] As a result, in the case of camellia extract extracted by existing extraction methods, IC 50 While the value is 443.17 μg / mL, the camellia extract extracted by the high-temperature and high-pressure extraction method of the present invention has an IC 50 The value was 262.99 μg / mL, indicating an increase in antioxidant activity of approximately 40.7%.

[0068] Furthermore, in the case of evergreen oak extract extracted by existing extraction methods, IC 50 While the value is 340.70 μg / mL, the Japanese evergreen oak extract extracted by the high-temperature and high-pressure extraction method of the present invention has an IC 50 The value was 250.34 μg / mL, indicating an increase in antioxidant activity of approximately 26.5%.

[0069] Furthermore, in the case of Cinnamomum japonicum extract extracted by existing extraction methods, IC 50On the other hand, the camellia extract extracted by the high-temperature and pressure extraction method of the present invention showed an IC 50 value of 163.82 μg / mL, indicating a numerical value with an antioxidant activity increased by 52.7%.

[0070] <1-2>ABTS radical scavenging ability

[0071] The ABTS radical scavenging ability can measure the radical scavenging activities of both hydrophilic and hydrophobic samples and has the advantage of a wide application range. In this experiment, it was measured by modifying the method of Jeong et al. Specifically, a 7 mM ABTS solution dissolved in distilled water and 2.45 mM potassium persulfate were mixed in a 1:1 ratio and left in the dark for 12 to 16 hours. The radical stock solution was diluted with PBS (pH 7.4) so that the absorbance value was 0.70 ± 0.02. 200 μL of the samples prepared at different concentrations (50 to 1,000 μg / mL) were added to 1000 μL of the diluted solution, and after reacting for 15 minutes in the dark, the reaction was carried out. Thereafter, the absorbance was measured at 517 nm using an HT multi-detection microplate reader (Synergy HT, BIO-TEX, Winooski, VT, USA). The scavenging ability (IC 50 ) of the extract against ABTS was expressed as the concentration required to reduce the absorbance of the control group using only the solvent by 50%.

[0072] TIFF2026079819000005.tif15165

[0073] As a result, in the case of the camellia extract extracted by the existing extraction method, the IC 50 value was 463. However, in the case of the camellia extract extracted by the high-temperature and pressure extraction method of the present invention, the IC 50 value was 260.09 μg / mL, indicating a numerical value with an antioxidant activity increased by about 43.8%.

[0074] Furthermore, in the case of evergreen oak extract extracted by existing extraction methods, IC 50 While the value is 199.31 μg / mL, the Japanese evergreen oak extract extracted by the high-temperature and high-pressure extraction method of the present invention is IC 50 The value was 117.54 μg / mL, indicating an increase in antioxidant activity of approximately 41.0%.

[0075] Furthermore, in the case of Cinnamomum japonicum extract extracted by existing extraction methods, IC 50 While the value is 57.69 μg / mL, the Cinnamomum japonicum extract extracted by the high-temperature and high-pressure extraction method of the present invention is IC 50 The value was 41.07 μg / mL, indicating a 28.8% increase in antioxidant activity.

[0076] [Table 3]

[0077] <Experimental Example 2>

[0078] Total polyphenol and total flavonoid content of the extract

[0079] <2-1> Total polyphenol content analysis

[0080] Polyphenols are aromatic alcohol compounds found in plants that have two or more phenolic groups as active groups in a single molecule. They have diverse structures and molecular weights, and because they contain hydroxyl groups, they readily bind to a variety of compounds, including water-soluble proteins. Polyphenols have diverse physiological functions such as anti-cancer, anti-inflammatory, and antioxidant properties, and these functions vary depending on the content and components. In this experiment, the total polyphenol content of camellia, evergreen oak, and Japanese cinnamon extracts was measured using extraction methods.

[0081] Total polyphenol content was measured using the Folin-Ciocalteu method as a reference (Ainsworth, EA, & Gillespie, KM, Nat. Protoc., 2(4), 875-877(2007)). In a simplified manner, the extract was prepared to a concentration of 500 μg / mL. Then, 500 μL of the prepared sample was sequentially mixed with 500 μL of 0.2 M Folin-Ciocalteu's phenol reagent and 500 μL of 2% sodium carbonate aqueous solution (w / v), and the mixture was reacted in a dark room at room temperature for 30 minutes. Subsequently, the absorbance was measured at 750 nm using an HT multi-detection microplate reader. After creating calibration curves using gallic acid diluted to different concentrations as standard substances, the gallic acid (GAE) mg / g equivalent was converted using the calibration curves.

[0082] As a result, the total polyphenol content of the camellia extract, evergreen oak extract, and Japanese cinnamon extract extracted by the high-temperature and high-pressure extraction method of the present invention increased by approximately 87.1%, 32.3%, and 18.5%, respectively, compared to existing extraction methods (see Table 4).

[0083] <2-2> Total flavonoid content analysis

[0084] Flavonoids, which exist in various forms in plants, belong to the polyphenol group and have very high anti-inflammatory, anti-cancer, anti-obesity, and antioxidant effects. In this experiment, the total flavonoid content of camellia, evergreen oak, and Japanese cinnamon extracts was measured using extraction methods.

[0085] For total flavonoid content analysis, the extract was prepared to a concentration of 1000 μg / mL. Then, 500 μL of the prepared sample was sequentially mixed with 1.5 mL of methanol, 100 μL of 1 M potassium acetate, and 1.4 mL of distilled water. After reacting at room temperature for 40 minutes, the absorbance was measured at 415 nm using an HT multi-detection microplate reader. Calibration curves were created using diluted quercetin (QUE) as a standard substance, and the quercetin (QUE) mg / g equivalent was then calculated.

[0086] As a result, the total flavonoid content of the camellia extract, evergreen oak extract, and Japanese cinnamon extract extracted by the high-temperature and high-pressure extraction method of the present invention increased by approximately 20.6%, 157.3%, and 880.7%, respectively, compared to existing extraction methods (see Table 4).

[0087] [Table 4]

[0088] <Experimental Example 3>

[0089] Component analysis using LC-MS / MS analysis of extracts

[0090] To quantitatively analyze the active ingredients contained in the camellia extract, evergreen oak extract, and Japanese cinnamon extract prepared according to <Example 1> and <Comparative Example 1> above, nine types of polyphenols (4-hydroxybenzoic acid, coumaric acid, rutin, naringenin, gallic acid, protocatechuic acid, chlorogenic acid, catechin, and tannic acid) were analyzed using HPLC-MS / MS (AB SCIEX 4000 Q Trap LC / MS / MS System, Shimadzu LC 20A System). A 10 μL sample was injected and analyzed using a C18 column (Gemini 3 μm, C18 110A 50 mm × 2.0 mm) under column oven (40 °C) conditions with an autosampler (15 °C). Analysis was performed using a turbo ion spray in both anionic and cation modes, with water (0.1% formic acid) (A) and acetonitrile (0.1% formic acid) (B) as the mobile phases. The anion mode was analyzed under isocratic conditions with mobile phase (B) as the reference: 0-0.5 min: B (20-20%), 0.5-2 min: B (20-80%), 2-2.5 min: B (80-80%), 2.5-2.6 min: B (80-20%), 2.6-6 min: B (20-20%), and 0-3 min: B (30-30%). The cation mode was analyzed under conditions of 0-0.2 min: B (30-60%), 0.2-2 min: B (60-60%), 2-2.1 min: B (60-30%), and 2.1-4 min: B (30-30%).

[0091] The results are shown in detail in Table 5.

[0092] Analysis of polyphenols using LC-MS / MS revealed that, compared to existing extraction methods, the camellia extract extracted by the high-temperature and high-pressure extraction method of the present invention showed increases of 380% in hydroxybenzoic acid, 217.7% in coumaric acid, 573.2% in rutin, 139.3% in naringenin, 787.4% in gallic acid, 814.1% in protocatechuic acid, 280.2% in catechin, and 9,543% in tannic acid.

[0093] Compared to existing extraction methods, the Japanese evergreen oak extract obtained by the high-temperature and high-pressure extraction method of the present invention showed increases of 149.6% in hydroxybenzoic acid, 3,720% in coumaric acid, 453.8% in naringenin, 1,710.2% in gallic acid, 48,483.1% in protocatechuic acid, 63.9% in catechin, and 508% in tannic acid.

[0094] Compared to existing extraction methods, the Cinnamomum japonicum extract extracted by the high-temperature and high-pressure extraction method of the present invention showed increases in hydroxybenzoic acid content by 687.7%, rutin by 590.6%, naringenin by 131.6%, gallic acid by 12,214.5%, and protocatechuic acid by 363.3%. In particular, chlorogenic acid, catechin, and tannic acid were not detected in extracts obtained using existing extraction methods, but were detected at 2,362.70 μg / g, 4.16 μg / g, and 2,800 μg / g, respectively, in the Cinnamomum japonicum extract extracted by the high-temperature and high-pressure extraction method of the present invention.

[0095] [Table 5]

[0096] For reference, gallic acid is a powerful antioxidant, and protocatechuic acid, a type of phenolic acid found in green tea, is known to protect the skin from oxidative stress caused by free radicals as a major metabolite of antioxidant polyphenols. Chlorogenic acid, a naturally occurring polyphenol, is one of the powerful antioxidants that alleviates oxidative stress on cells and prevents the development of various symptoms and diseases caused by it. Catechin is known to affect human skin cells involved in aging, suppressing wrinkle formation and improving skin moisture and elasticity, while tannic acid has been reported to suppress elastase activity in a concentration-dependent manner and is known as a substance that suppresses wrinkle formation.

[0097] Therefore, compared to existing extraction methods, using the high-temperature and high-pressure extraction method of the present invention significantly increases the extracted content of protocatechuic acid and chlorogenic acid, along with gallic acid, which are powerful antioxidants. It also significantly increases the content of catechin and tannic acid, which are effective in suppressing wrinkle formation. As a result, the extract obtained using the high-temperature and high-pressure extraction method of the present invention can be usefully used as a material with functional properties such as wrinkle improvement, skin moisturizing, and whitening.

[0098] Dosage form example 1: Softening lotion (skin lotion)

[0099] As shown in the table below, the softening lotion was manufactured using conventional methods.

[0100] [Table 6]

[0101] Dosage form example 2: Nourishing lotion (milk lotion)

[0102] As shown in the table below, the nourishing lotion was manufactured using conventional methods.

[0103] [Table 7]

[0104] Dosage form example 3: Nutritional cream

[0105] As shown in the table below, the nutritional cream was manufactured using conventional methods.

[0106] [Table 8]

[0107] Dosage form example 4: Massage cream

[0108] As shown in the table below, the massage cream was manufactured using conventional methods.

[0109] [Table 9]

[0110] Dosage form example 5: Pack

[0111] As shown in the table below, the packs were manufactured using conventional methods.

[0112] [Table 10]

[0113] As described above, the present invention has been explained primarily in terms of its preferred embodiments. A person with ordinary skill in the art to which the present invention pertains should understand that the present invention can be embodied in modified forms that do not depart from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered in an explanatory rather than restrictive manner. The scope of the present invention is defined not by the foregoing description but by the claims, and all differences within an equivalent scope are considered to be within the scope of the present invention.

Claims

1. a) A step in which the raw materials are washed and dried and then crushed to a size of 0.3 to 0.7 mm, b) A step of extracting the extract by adding high-temperature steam under a pressure of 8 to 10 bar, c) A method for producing an extract with increased polyphenol and flavonoid content, comprising the step of filtering the extract and then concentrating it.

2. The method for producing an extract with increased polyphenol and flavonoid content according to claim 1, characterized in that the raw material is one selected from the group consisting of camellia, evergreen oak, Japanese cinnamon, and edible mushrooms.

3. The method for producing an extract with increased polyphenol and flavonoid content according to claim 1, characterized in that the polyphenol is one or more selected from the group consisting of 4-hydroxybenzoic acid, coumaric acid, rutin, naringenin, gallic acid, protocatechuic acid, chlorogenic acid, catechin, and tannic acid.

4. An extract having increased polyphenol and flavonoid content, produced by the method described in any one of claims 1 to 3.

5. A cosmetic composition comprising the extract of claim 4 as an active ingredient.

Images