Tomato extract with excellent antioxidant activity and tomato juice comprising the same

Abstract

The present invention relates to a tomato extract having excellent antioxidant activity and tomato juice containing the same. More specifically, the present invention relates to a tomato extract having excellent antioxidant activity and improved anti-aging and anti-inflammatory effects, tomato juice containing the same, and a method for manufacturing the same.

Description

Technology Field

[0001] The present invention relates to a tomato extract having excellent antioxidant activity and tomato juice containing the same. More specifically, the present invention relates to a tomato extract having excellent antioxidant activity and improved anti-aging and anti-inflammatory effects, tomato juice containing the same, and a method for manufacturing the same. Background Technology

[0003] Tomato (Lycopersicon esculentum) is an annual crop of the Solanaceae family that is mainly cultivated in temperate regions. It is one of the world's major edible crops, consumed by many people around the world as a raw ingredient, but also processed and consumed in various other forms such as juice, sauces, and stews. It is also a representative fruit and vegetable that is cultivated nationwide in Korea, where the climate and soil conditions are suitable.

[0004] In traditional Korean medicine, the tomato is referred to as *beonga* or *nyeongam*. Its fruit is primarily consumed for its ability to promote digestion, strengthen the stomach, and stimulate the secretion of bodily fluids, while in folk medicine, it has been used as a remedy for hypertension, diabetes, and night blindness. Tomatoes contain various nutrients, including vitamins A, B, C, E, and K, as well as minerals, organic acids, amino acids, and sugars, along with bioactive substances such as lycopene and β-carotene. In particular, the lycopene found in tomatoes exhibits effects such as oxidation inhibition, antioxidant activity, anti-aging benefits, and inhibition of prostate cancer, and related studies confirming these effects have been reported.

[0005] It is known that the content and activity of the functional components of tomatoes, as described above, vary depending on the variety, lineage, cultivation environment, and post-harvest management. Furthermore, studies on the carotenoid components of tomatoes have suggested that lycopene and β-carotene are associated with fruit color, fruit shape, acidity, and sugar content. Additionally, quantitative analysis of lycopene, β-carotene, and xanthophyll in ripe tomato lines and varieties revealed that xanthophyll was most abundant in the variety 'Delice', and a significant correlation was observed between the lycopene and xanthophyll content. An investigation into the differences in ascorbic acid, lycopene, β-carotene, and α-carotene content between ripe tomato varieties and processed tomato products revealed that there are differences in the levels of bioactive substances depending on the variety and the processed product. Many studies have reported that the lycopene content of fresh tomatoes ranges from 1.82 to 11.19 mg / 100 g, while that of tomato juice ranges from 6.93 to 42.74 mg / 100 g, suggesting that consuming juice may be more efficient for lycopene intake.

[0006] As such, while numerous studies on the efficacy of tomatoes have been conducted both domestically and internationally, research on tomato juice—an easily consumable beverage with high antioxidant properties—is currently lacking.

[0008] As background technology for the present invention, Korean Registered Patent Publication No. 10-0865132 describes a method for manufacturing cherry tomato juice and cherry tomato juice manufactured thereby. The problem to be solved

[0010] The objective of the present invention is to provide a tomato extract having excellent antioxidant and anti-aging activities.

[0011] Another objective of the present invention is to provide a method for producing a tomato extract having excellent antioxidant and anti-aging activities.

[0012] Another objective of the present invention is to provide tomato juice containing a tomato extract having excellent antioxidant and anti-aging activities.

[0013] Another objective of the present invention is to provide a method for producing tomato juice containing a tomato extract having excellent antioxidant and anti-aging activities.

[0014] Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims, and drawings. means of solving the problem

[0016] According to one aspect, a method for producing a tomato extract having excellent antioxidant activity is provided, comprising: Ii) a washing step of washing a tomato; I-ii) a grinding step of grinding the tomato washed in step Ii); and I-iii) a step of producing a tomato extract by boiling the tomato ground in step I-ii) in water at a temperature of 80 to 90°C for 20 to 40 minutes.

[0017] According to one embodiment, after step I-iii), a step of filtering the water bath extract may be further included.

[0019] According to another aspect, a tomato extract having excellent antioxidant activity is provided, prepared by the method described herein.

[0020] According to one embodiment, the anti-aging activity of the tomato extract can be improved.

[0021] According to one embodiment, the anti-inflammatory effect of the tomato extract can be improved.

[0022] According to one embodiment, the tomato extract may have an increased reducing sugar content.

[0024] According to another aspect, tomato juice comprising the tomato extract described herein is provided.

[0026] According to another aspect, a method for producing tomato juice is provided, comprising: II-i) a step of preparing a tomato extract produced by the method of production described herein; II-ii) a first mixing step of preparing a mixture by adding salt, sugar, and purified water to the tomato extract of step II-i); II-iii) a grinding step of grinding the mixture of step II-ii); II-iv) a second mixing step of re-mixing the ground mixture of step II-iii); and II-v) a packaging step of packaging the re-mixed mixture of step II-iv) in a packaging container.

[0027] According to one embodiment, in step II-ii), a mixture may be prepared by adding 0.1 to 0.5 parts by weight of salt and 0.5 to 1 part by weight of sugar to 100 parts by weight of tomato extract.

[0028] According to one embodiment, the method for manufacturing tomato juice of the present invention may further include: II-vi) a sterilization step of placing the packaged product of step II-v) into a metal case and sterilizing it at a temperature of 80 to 90°C for 20 to 30 minutes; and II-vii) a step of drying the sterilized packaged product of step II-vi). Effects of the invention

[0030] According to one embodiment, the tomato extraction method of the present invention includes a water bath extraction step, thereby enabling the production of a tomato extract with excellent antioxidant activity.

[0031] According to one embodiment, the tomato extraction method of the present invention includes a water bath extraction step, and can produce a tomato extract having excellent anti-aging activity compared to an extract that is not extracted by water bath.

[0032] According to one embodiment, the tomato extraction method of the present invention includes a water bath extraction step, and can produce a tomato extract with excellent anti-inflammatory effects compared to an extract that is not extracted by water bath.

[0033] According to one embodiment, the tomato extraction method of the present invention includes a water bath extraction step, and can produce a tomato extract having an increased reducing sugar content compared to an extract that is not extracted by water bath.

[0034] According to one embodiment, tomato juice containing the tomato extract of the present invention may have antioxidant, anti-aging activity, and anti-inflammatory effects after undergoing a double-boiler extraction process of tomatoes.

[0035] According to one embodiment, the method for producing tomato juice of the present invention can efficiently produce tomato juice with improved antioxidant, anti-aging, and anti-inflammatory activities. Brief explanation of the drawing

[0037] Figure 1 is a graph showing the results of measuring the polyphenol and flavonoid content of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2. FIG. 2 is a graph showing the results of measuring the reducing sugar and water-soluble protein content of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2. Figure 3 is a graph showing the results of measuring the DPPH radical scavenging rate according to the tomato extract concentration of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2. Figure 4 is a graph showing the results of measuring SOD-like activity according to the concentration of tomato extracts of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2. Figure 5 is a graph showing the results of evaluating the XO inhibitory activity according to the concentration of tomato extracts of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2. Figure 6 is a graph showing the results of evaluating the collagenase inhibition rate according to the concentration of tomato extracts of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2. Figure 7 is a graph showing the results of evaluating the tyrosinase inhibitory activity according to the concentration of tomato extracts of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2. FIG. 8 is a graph showing the cell viability of RAW264.7 according to the tomato extract concentrations of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2. FIG. 9 is a graph showing the cell viability of HaCaT cells according to the concentration of tomato extract of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2. FIG. 10 is a graph showing the results of evaluating the NO production inhibitory activity according to the concentration of tomato extracts of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2. Specific details for implementing the invention

[0038] The objects, advantages, and features of the present invention will become more apparent from the following detailed description and embodiments associated with the attached tables and drawings.

[0039] Prior to this, terms and words used in this specification and claims should not be interpreted or evaluated in their ordinary and dictionary meanings, but should be interpreted in a meaning and concept consistent with the technical spirit of this disclosure, based on the principle that the inventor can appropriately define the concept of the terms to best describe his invention.

[0040] The terms used herein are merely for describing specific embodiments and are not intended to limit the disclosure. The singular expression includes the plural expression unless the context clearly indicates otherwise.

[0041] In this specification, terms such as 'comprising' or 'having' are intended to indicate the existence of the features, numbers, steps, actions, components, parts, or combinations thereof described in the specification, and should be understood as not excluding in advance the existence or addition of one or more other features, numbers, steps, actions, components, parts, or combinations thereof.

[0042] In this specification, when a part is described as "comprising" a certain component, it means that, unless specifically stated otherwise, it does not exclude other components but may include additional components.

[0043] The contents of the present invention will be explained in more detail below in the following examples. However, the scope of the present invention is not limited to the following examples but includes variations of equivalent technical concepts.

[0045] According to one aspect, a method for preparing a tomato extract having excellent antioxidant activity according to the present invention comprises: Ii) a washing step of washing a tomato; I-ii) a grinding step of grinding the tomato washed in step Ii); and I-iii) an extract preparation step of preparing a water bath extract by boiling the tomato ground in step I-ii) in water at a temperature of 80 to 90°C for 20 to 40 minutes.

[0046] The tomato of the present invention may be a common large-fruited tomato or a cherry tomato, and a common large-fruited tomato may be more suitable for the production of the tomato extract with excellent antioxidant activity of the present invention.

[0048] Step IIi) is a tomato washing step, which is a step of washing the tomatoes thoroughly to remove foreign substances from the surface before crushing them. Although not limited thereto, a washing step in which ripe tomatoes are washed once under running water, the stems are removed, and the tomatoes are rinsed two more times may be suitable.

[0049] Although not limited thereto, a step of removing moisture from the washed tomatoes may be further included after step Ii) above.

[0051] Step I-ii) is a grinding step for grinding the tomatoes washed in Step Ii). A blender may be used for the grinding step, and the grinding time can be adjusted according to the amount of tomatoes.

[0053] Step I-iii) is a step for preparing a tomato extract, wherein the tomatoes crushed in Step I-ii) are boiled in water at a temperature of 80 to 90°C for 20 to 40 minutes.

[0054] Although not limited thereto, the above-mentioned water bath temperature may be 80 to 90°C, which is suitable for improving the antioxidant activity, anti-aging activity, and anti-inflammatory effect of the tomato extract, and 85 to 90°C, which is more suitable. The above-mentioned water bath time may be 20 to 40 minutes, which is suitable for improving the antioxidant activity, anti-aging activity, and anti-inflammatory effect of the tomato extract, and 20 to 30 minutes, which is more suitable.

[0055] If the above-mentioned range of water bath temperature and time is exceeded, the improvement in the antioxidant activity, anti-aging activity, and anti-inflammatory effects of the tomato extract may be minimal, the taste of the juice may not be refreshing when consumed after preparation, and the color of the tomato extract may change to brown, causing the loss of the tomato's natural color and reducing the palatability of the tomato juice containing the tomato extract.

[0057] Although not limited thereto, the method for preparing a tomato extract according to the present invention may further include a step of filtering the tomato water bath extract after step I-iii). The filtration step may use a filtration method commonly used in the preparation of fruit or vegetable juices and extracts, and is not particularly limited.

[0059] According to another aspect, a tomato extract having excellent antioxidant activity is provided, which is produced by the method for producing a tomato extract of the present invention.

[0060] The tomato extract of the present invention may have improved antioxidant activity compared to a tomato extract extracted by other extraction methods that are not water bath extraction. For example, the antioxidant activity may be improved by improving DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity, the antioxidant activity may be improved by improving SOD (Superoxide dismutase)-like activity, and the antioxidant activity may be improved by improving XO (Xanthine oxidase) inhibitory activity.

[0061] DPPH is a relatively stable free radical used in the measurement of electron-donating capacity. Since it is reduced by sulfur-containing amino acids such as cysteine ​​and glutathione, as well as by ascorbic acid and BHA, it is useful for measuring antioxidant activity from various extracts. The DPPH radical scavenging method is a representative method for measuring the antioxidant capacity of plant extracts and foods, and it allows for easy measurement of antioxidant activity by utilizing the fact that antioxidant active substances decolorize as they scavenge DPPH radicals.

[0062] SOD exists in all oxygen-consuming organisms and is a representative reactive oxygen species inhibitor that acts as a defense mechanism against highly harmful superoxide anion radicals (O₂) within the body. 2- Reaction that converts ·) into hydrogen peroxide (H2O2) 2 O2 2-It is an enzyme that catalyzes the reaction (+2H → H2O2 + O2). Hydroxy radicals generated in the body are reduced into harmless water and oxygen by hydrolytic enzymes such as peroxidase and catalase, thereby eliminating superoxide anion radicals. Therefore, it can suppress aging and various chronic diseases caused by reactive oxygen species, such as cardiovascular disease, cancer, and dementia. It can also be utilized as an anti-inflammatory agent for degenerative diseases, including arthritis and rheumatism, or as an ingredient in anti-aging cosmetics. Furthermore, in the agricultural sector, it can enhance plants' resistance to environmental stress.

[0063] XO is involved in the metabolism of purines, pyrimidines, pteridines, aldehydes, and heterocyclic compounds in the body, and generates uric acid by utilizing molecular oxygen from xanthine or hypoxanthine as an electron acceptor. When crystalline forms increase in the plasma, they are deposited in peri-joint tissues and kidneys, causing gout and kidney disease accompanied by inflammation and pain. Therefore, XO activity inhibitors are used as gout treatments to reduce blood uric acid levels.

[0065] Although not limited thereto, the tomato extract of the present invention may have improved anti-aging activity compared to tomato extracts extracted by other extraction methods that are not water bath extraction. For example, the collagenase inhibitory effect may be improved, thereby improving anti-wrinkle and anti-aging effects, and the tyrosinase inhibitory effect may be improved, thereby improving whitening and anti-aging effects.

[0066] External factors, such as stress caused by exposure to harmful environments and an increase in reactive oxygen species due to UV irradiation, can also accelerate the destruction of the skin matrix, leading to the formation of wrinkles. While there are various types of collagen-degrading enzymes, Matrix Metalloproteinases (MMPs), which are representatively present in the dermis, show increased expression when exposed to reactive oxygen species and UV rays. MMPs are zinc-containing metalloproteinases, including collagenase, gelatinase, and elastase, which function to decompose extracellular and basement membrane components. Among these, excessive exposure to UV rays causes skin collagenase (MMP-1) to significantly destroy dermal collagen. Since this plays a crucial role in photoaging, reducing collagenase activity is critical for the decline in skin elasticity and the formation of wrinkles. Collagen, which accounts for more than 90% of the skin's dermis, forms the majority of the organic matter in tendons, bones, and teeth, in addition to the skin. In the case of the skin, it has the function of inducing and protecting the mechanical firmness, resistance, tissue strength, and differentiation of skin cells. However, since a decrease in collagen can lead to a decline in the production of organic substances in tendons and bones, it is closely related not only to beauty but also to the bone health of the human body.

[0067] Tyrosinase is the only enzyme involved in melanin production; when skin is exposed to ultraviolet rays, L-tyrosine is oxidized to L-3,4-dihydroxyphenylalanine (DOPA) and DOPA quinone, then further to DOPA chrome, 5,6-hydroxyindole, and indole-5,6-quinone, and via indole-5,6-quinone, it utilizes a wide range of phenolic compounds as substrates, such as Cu 2+It is an enzyme containing [it]. Therefore, if melanin synthesis is inhibited by inhibiting the activity of tyrosinase, a whitening effect on pigmentation such as melasma, freckles, and senile erythema can be directly confirmed.

[0069] Although not limited thereto, the tomato extract of the present invention may have an improved anti-inflammatory effect compared to a tomato extract obtained by other extraction methods that are not water bath extraction. For example, the anti-inflammatory effect may be improved by improving the activity that inhibits NO (Nitric oxide) production.

[0070] NO is produced by the catalysis of nitric oxide synthase (NOS) to regulate acute and chronic inflammatory responses; in particular, iNOS is known as a regulatory molecule involved in inflammatory responses that plays an important role both physiologically and pathologically. While appropriate concentrations of NO are physiologically necessary for smooth muscle relaxation, inhibition of platelet aggregation, immune modulation, vasodilation, and neurotransmission, chronic production of NO acts as an inflammatory factor and becomes a major cause of various types of chronic inflammatory diseases.

[0072] Although not limited thereto, the tomato extract of the present invention contains a large amount of polyphenols and flavonoids, which are representative beneficial components of plants, and may have excellent effects such as antioxidant, anticancer, anti-inflammatory, and antibacterial properties.

[0073] Polyphenols are representative secondary metabolites of plants and are multifunctional substances widely present in plant tissues. Possessing one or more hydroxyl groups within their molecules, they act as hydrogen donors and are known to be involved in antioxidant, antibacterial, anti-allergic, and anticancer activities by contributing to the resonance stabilization of the phenol structure. Flavonoids are phenolic compounds synthesized by plants that are pigment compounds exhibiting yellow, pale yellow, and reddish-purple colors. In plants, they mostly exist in the form of glycosides bound to sugars and are known to possess various physiological functions, such as anti-cardiovascular, anticancer, and anti-inflammatory effects, by effectively scavenging reactive oxygen species.

[0075] Although not limited thereto, the tomato extract of the present invention can be applied to drinkable foods such as juices, drinks, and juices, and can be applied to processed tomato foods processed by general processing methods, but it may be most suitable to be included in juice.

[0077] According to another aspect, the method for producing tomato juice of the present invention comprises: II-i) a step of preparing a tomato extract produced by the method of production described herein; II-ii) a first mixing step of preparing a mixture by adding salt, sugar, and purified water to the tomato extract of step II-i); II-iii) a grinding step of grinding the mixture of step II-ii); II-iv) a second mixing step of re-mixing the ground mixture of step II-iii); II-v) a packaging step of packaging the re-mixed mixture of step II-iv) into a packaging container; II-vi) a sterilization step of placing the packaged product of step II-v) into a metal case and sterilizing it at a temperature of 80 to 90°C for 20 to 30 minutes; and II-vii) a step of drying the sterilized packaged product of step II-vi).

[0079] The tomato juice produced by the above manufacturing method may be produced in a liquid form that is drinkable, such as juice and drink.

[0081] Step II-i) is a step of preparing a tomato extract prepared by the method for preparing a tomato extract of the present invention. The method for preparing a tomato extract may include a washing step, a grinding step, and an extract preparation step of preparing a water bath extract by boiling the ground tomatoes in water at a temperature of 80 to 90°C for 20 to 40 minutes.

[0082] Although not limited thereto, the above tomato extract is extracted by water bath according to the method for preparing tomato extract of the present invention, and compared to tomato extracts extracted by other extraction methods that are not water bath extracted, it may have superior antioxidant activity and improved anti-aging and anti-inflammatory effects.

[0084] Step II-ii) is a first mixing step in which salt and sugar are added to improve the sweetness, umami, and palatability of the tomato juice being produced, and purified water is added to produce the juice to prepare the mixture.

[0085] Although not limited thereto, the tomato extract, salt, and sugar in step II-ii) above may be added in an amount of 100 parts by weight of tomato extract, 0.1 to 0.5 parts by weight of salt, and 0.5 to 1 part by weight of sugar to improve the sweetness, umami, and palatability of the tomato juice, and may be more suitable to be 100 parts by weight of tomato extract, 0.1 parts by weight of salt, and 0.6 parts by weight of sugar.

[0086] Although not limited thereto, in step II-ii) above, adding purified water in an amount of 4 to 10 parts by weight may be suitable for improving the sweetness, umami, and palatability of the tomato juice, adding it in an amount of 4 to 7 parts by weight may be more suitable, and adding it in an amount of 4 parts by weight may be even more suitable.

[0088] Step II-iii) is a grinding step for grinding the mixture of Step II-ii). When preparing the tomato extract in Step II-i), the tomatoes are ground once, then salt, sugar, and purified water are added, and then ground a second time. As a result, when drinking the prepared tomato juice, the ground tomato particles are small, making it easy to swallow and easy to drink.

[0090] Step II-iv) is a secondary mixing step for remixing the ground mixture of Step II-iii). By remixing the ground mixture, the tomato extract, salt, sugar, and purified water can be further mixed to improve palatability.

[0092] Step II-v) is a packaging step in which the mixture remixed in Step II-iv) is packaged in a packaging container. Although not limited thereto, the packaging container may be in the form of an opaque plastic bag manufactured to be drinkable.

[0094] Step II-vi) is a sterilization step in which the packaged product from Step II-v) is placed in a metal case and sterilized at a temperature of 80 to 90°C for 20 to 30 minutes, which is a process to enable the manufactured tomato juice product to be stored for a long period. The sterilization temperature and time can be determined so that the quality of the manufactured tomato juice product can be maintained fresh.

[0096] After the above sterilization step, a cooling step may be further included. By cooling within a short period of time after the heat-applied sterilization process, changes in the quality and color of the tomato juice caused by residual heat remaining after sterilization can be prevented.

[0098] Step II-vii) is a step of drying the sterilized packaged product from Step II-vi). Through this drying step, residual heat remaining after the sterilization step can be removed from the packaged product, and moisture, etc., from the packaging container can be removed, making it easier to store.

[0099] Although not limited thereto, the step of cooling the product after the above sterilization step may be performed simultaneously with the drying step, and the drying step may be the cooling step.

[0101] The present invention will be described in more detail below with reference to preferred embodiments. However, it will be obvious to those skilled in the art that these embodiments are intended to explain the invention more specifically and that the scope of the invention is not limited by them.

[0103] Examples

[0104] Ripe tomatoes ("One Top" variety tomatoes provided by "Bunong") were washed and ground using a blender (Waring, HGB2WTS3, USA), then boiled in a water bath at 90°C for 30 minutes to prepare a water bath extract, which was then filtered through filter paper (Whatman No. 2, England). The prepared extract was named Example or LBW (Boiling water extract from Lycopersicon esculentum).

[0106] Comparative example

[0107] Comparative Example 1

[0108] The stems of ripe tomatoes were removed and the mixture was ground using a blender. Distilled water equivalent to 10 times the weight of the sample was placed in a round-bottom flask equipped with a reflux condenser, and the mixture was extracted in a 60°C water bath for 3 hours. This process was repeated 3 times to prepare a water extract using a reflux extractor. Each extract was filtered through filter paper (Whatman No. 2, England) and concentrated under reduced pressure to approximately 1 / 10 using a rotatory vacuum evaporator. The prepared extract was named Comparative Example 1 or LRW (Reflux water extract from Lycopersicon esculentum).

[0110] Comparative Example 2

[0111] The fresh juice extract was prepared by grinding fully ripened tomatoes with the stems removed using a blender, filtering out only the juice using a sieve, and filtering it through filter paper. The prepared extract was named Comparative Example 2 or LRJ (Raw juice extract from Lycopersicon esculentum).

[0113] Experimental Example

[0114] 1. Analysis of beneficial component content

[0115] 1-1. Analysis of Polyphenol Compound Content

[0116] The total content of polyphenol compounds contained in ripe tomato extracts was measured using the Folin-Denis (AOAC, 2005) method. Tomato extracts obtained under different conditions were dissolved in distilled water at a 10% concentration. Then, 0.2 mL of the tomato extract sample diluted to a specific concentration was mixed with 1.8 mL of distilled water and 0.2 mL of Folin-Denis's phenol reagent, and reacted at room temperature for 3 minutes. Subsequently, 0.4 mL of saturated Na2CO3 and 1.4 mL of distilled water were added and mixed, followed by a reaction at room temperature for 1 hour. The absorbance was then measured at 725 nm using a UV / VIS spectrophotometer. Standard curves were prepared using tannic acid to obtain solutions with final concentrations of 0, 25, 50, 100, 250, and 500 µg / mL. These were measured using the same method as above to construct the standard curves, which were then used to calculate the total content of polyphenol compounds contained in the tomato extracts.

[0118] 1-2. Analysis of Flavonoid Compound Content

[0119] The flavonoid compound content was measured by modifying the method of Nieva Moreno et al. (2000). Tomato extract was dissolved in 80% ethanol as a solvent and diluted to a specific concentration. Then, 0.1 mL of 10% aluminum nitrate, 0.1 mL of 1 M potassium acetate, and 4.7 mL of 80% ethanol were added to 0.1 mL of the sample and reacted at 25°C for 40 minutes. The absorbance was then measured at 415 nm using a UV / VIS spectrophotometer. For quantification, quercetin was used to obtain final concentrations of 0, 25, 50, 100, 250, and 500 µg / mL. The samples were measured using the same method as above, and a standard curve was constructed. This curve was then used to calculate the flavonoid compound content contained in the tomato extracts obtained under different conditions.

[0120] Table 1 is a table showing the results of measuring the polyphenol and flavonoid content of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2.

[0121] Extract Compound contents (mg / g) Polyphenol Flavonoid LRW (Comparative Example 1) 12.78 ± 0.35 c 7.60 ± 0.4 b LRJ (Comparative Example 2) 13.57 ± 0.8 b 6.93 ± 0.26 c LBW (Example) 15.40 ± 0.26 a 8.76 ± 0.43 a

[0122] Figure 1 is a graph showing the results of measuring the polyphenol and flavonoid content of the Example, Comparative Example 1, and Comparative Example 2.

[0123] Referring to Table 1 and Figure 1, tomato extracts extracted by different methods contained large amounts of polyphenols and flavonoids exhibiting antioxidant and anti-inflammatory effects, and compared to Comparative Example 1 and Comparative Example 2, the example showed the highest content of polyphenol compounds and flavonoids.

[0125] 1-3. Measurement of Reducing Sugar Content

[0126] The reducing sugar content was determined by diluting the tomato extract to a specific concentration and filtering it to use as the sample solution. Following the Somogyi-Nelson (Nelson, 1944) method, 0.5 mL of mixed reagent (A : B = 25 : 1, A; d3H2O 1 L in anhydrous Na2HPO4 25 g, C4H4O6KNa·4H2O 25 g, Na2HCO3 20 g, anhydrous Na2SO4 200 g; B; H2O 200 mL in CuSO4·5H2O 30 g, concentrate H2SO4 4 drop) was added to 1 mL of the sample solution, heated for 20 minutes, and then cooled to obtain Solution C (total 500 mL store at 37℃ / day - (NH4)6Mo7O 2425 g of 4H2O in 450 mL of d3H2O (including 1 mL of concentrate H2SO4 + 3 g of Na2HSO4·7H2O in 25 mL of d3H2O) was added and mixed. After the reaction, 5 mL of H2O was added, and the absorbance was measured using a spectrophotometer at 520 nm. To quantify reducing sugars, a standard curve was constructed using L-glucose, and the reducing sugar content was calculated using this curve.

[0128] 1-4. Measurement of Water-Soluble Protein Content

[0129] The total water-soluble protein content in each tomato extract was determined by diluting the sample to a specific concentration with distilled water and, according to the method of Lowry et al. (1951), placing 0.2 mL of the sample into a test tube and adding 1 mL of a mixed reagent (A : B = 50 : 1, A; 2% Na2CO3 in 0.1N NaOH, B; 1% C4H4KNaO6 in 0.5% CuSO45H2O), reacting at room temperature for 10 minutes. Subsequently, 0.1 mL of folin-ciocalteu's phenol reagent was added and reacted at room temperature for 30 minutes, after which the absorbance was measured at 750 nm using a spectrophotometer. The standard substance was bovine serum albumin, and the water-soluble protein content in each extract was calculated by measuring it in the same manner as above.

[0130] Table 2 is a table showing the results of measuring the reducing sugar and water-soluble protein content of the Example, Comparative Example 1, and Comparative Example 2.

[0131] Extract Compound contents (mg / g) Soluble protein Reducing sugar LRW (Comparative Example 1) 45.51 ± 1.06 a 57.40 ± 4.57 b LRJ (Comparative Example 2) 31.27 ± 0.92 b 44.43 ± 3.99 c LBW (Example) 44.50 ± 1.15 a 94.84 ± 1.54 a

[0132] FIG. 2 is a graph showing the results of measuring the reducing sugar and water-soluble protein content of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2.

[0133] Referring to Table 2 and Figure 2, the highest amount of reducing sugars was detected in the Examples compared to Comparative Examples 1 and 2 regarding reducing sugars extracted from tomatoes under different conditions, confirming that heating at high heat may be more suitable for extracting reducing sugars than heating with fresh juice at low heat. In the case of water-soluble proteins in tomatoes, Comparative Example 1 showed the highest content, but it was almost similar to the Examples.

[0134] When compared to the results (Lee et al., 2011) which showed that the reducing sugars in commercially available domestic juices were 32.90 mg / g for apple juice, 48.19 mg / g for grape juice, and 16.51 mg / g for orange juice, the extract in the example, which was heated in a water bath at 90°C for 30 minutes, contained approximately 1.9 to 5.7 times more reducing sugars than the commercially available juices. In addition, it contained approximately 2.8 times more reducing sugars than commercially available tomato juice, which contained 32.29 mg / g of reducing sugars. Consequently, it was confirmed that extracting tomatoes by heating at slightly higher temperatures may be a suitable method for extracting the most reducing sugars and water-soluble proteins.

[0136] 2. Evaluation of Antioxidant Activity

[0137] 2-1. Evaluation of DPPH Radical Scavenging Activity

[0138] The DPPH (2,2-diphenyl-1-picryl hydrazyl radical) scavenging activity of ripe tomato extracts obtained under different conditions was expressed by measuring the hydrogen-donating effect of DPPH using a modified method of Blois (1958). 50 µL of 0.2 mM DPPH solution (dissolved in 99.9% ethanol) was added to 100 µL of a diluted sample solution and reacted for 30 minutes, after which the absorbance was measured at 517 nm. The difference in absorbance between the tomato extract-added group and the non-added group was expressed as a percentage (%). To compare the DPPH radial scavenging activity, L-ascorbic acid, a natural antioxidant, was measured as a control using the same method as above and compared with the results of the tomato extract.

[0139] Table 3 shows the results of measuring the DPPH radical scavenging rate according to the tomato extract concentrations of the Examples, Comparative Example 1, and Comparative Example 2.

[0140] Table 4 shows the results of measuring the DPPH radical scavenging rate according to the concentration of the control group, L-ascorbic acid (AsA).

[0141] Con.(%) Extract LRW (Comparative Example 1) LRJ (Comparative Example 2) LBW (Example) 1.25 67.69±0.65 b 72.85±0.14 a 67.49±1.76 b 2.5 69.19±0.55 b 77.48±0.78 a 70.40±0.87 b 5 77.98±0.83 b 79.59±0.99 a 78.81±0.67 b 10 83.75±1.36 b 81.06±0.86 c 86.84±1.42 a

[0142] AsA (ug / mL) 62.5 125 250 500 89.41±0.96 89.73±1.27 91.15±1.25 93.73±0.40

[0143] Figure 3 is a graph showing the results of measuring the DPPH radical scavenging rate according to the tomato extract concentration of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2.

[0144] Referring to Tables 3 and 4 and Figure 3, the Example, Comparative Example 1, and Comparative Example 2 were lower than the control L-ascorbic acid (500 ug / mL, 93.73%) at a 10% concentration, but there was no significant difference (p<0.05). The Example, which is a water bath extract, was the highest at 86.84%, followed by Comparative Example 1 (LRW), which is a reflux extract, at 83.75%.

[0145] Meanwhile, according to the results of a comparison of the antioxidant effects of commercially available fruit and vegetable juices in Korea (Lee et al. (2011)), at a concentration of 5%, tomato juice showed 54.75%, apple juice 49.91%, and schisandra juice 38.73%. When comparing these results with the present study, the tomato extract showed approximately 1.4 times better antioxidant activity than commercially available tomato juice, and approximately 1.5 to 2 times higher DPPH radical scavenging effect than apple and schisandra juices.

[0146] In conclusion, the water-boiled tomato extract showed lower activity than the natural antioxidant L-ascorbic acid, but higher activity than commercially available fruit juices, confirming that it may be suitable for use as a material for necessary food or related businesses.

[0148] 2-2. Evaluation of SOD-like Activity

[0149] SOD-like activity was determined by measuring the oxidized amount of pyrogallol that catalyzes the conversion of harmful reducing oxygen species into H2O2 (hydrogen peroxide) according to the method of Marklund (1975), and expressing this as SOD-like activity. To 0.2 mL of tomato extract diluted to a certain concentration, 2.6 mL of tris-HCl buffer (50 mM tris[hydroxymethyl] amino-methane + 10 mM EDTA, pH 8.5) adjusted to pH 8.5 and 0.2 mL of 7.2 mM pyrogallol were added. After reacting at 25°C for 10 minutes, 0.1 mL of 1 N HCl was added to stop the reaction, and the amount of oxidized pyrogallol in the reaction solution was measured using a UV / VIS spectrophotometer at 420 nm. This was expressed as SOD-like activity by the difference in absorbance between the group with and without the tomato extract sample solution as a percentage (%), and as a control, L-ascorbic acid, a natural antioxidant, was measured in the same way and compared with the tomato extract of the example and comparative example.

[0150] Table 5 shows the results of measuring SOD-like activity according to the concentration of tomato extracts in the Examples, Comparative Example 1, and Comparative Example 2, measured as the pyrogallol automation inhibition rate.

[0151] Table 6 shows the results of measuring SOD-like activity according to the concentration of the control group, L-ascorbic acid (AsA).

[0152] Con.(%) Extract LRW (Comparative Example 1) LRJ (Comparative Example 2) LBW (Example) 1.25 - - 6.16±1.20 2.5 - - 8.64±1.00 5 3.86±0.42 - 17.49±1.96 10 25.76±1.1 - 35.76±0.68

[0153] AsA (ug / mL) 62.5 125 250 500 72.99±0.76 95.19±0.25 97.77±0.12 98.71±0.57

[0154] Figure 4 is a graph showing the results of measuring SOD-like activity according to the concentration of tomato extracts of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2.

[0155] Referring to Tables 5 and 6 and Figure 4, the example showed the highest SOD-like activity at 5% and 10%, while no similar activity was observed in Comparative Example 2. Although the example and Comparative Example 1 showed lower SOD-like activity than the purified natural antioxidant L-ascorbic acid, when compared to the results (Kim et al. (2004)) which showed effects of 3.2%, 10.3%, and 11.6% for the commercially available herbal medicines Polygonatum, Glycyrrhiza, and Angelica gigas, it can be confirmed that the heated tomato extract showed higher SOD-like activity than the commercially available herbal medicines.

[0156] In conclusion, since SOD is an enzyme involved in the defense mechanism of eliminating reactive oxygen species in the body, heated tomato reflux extract and double-boiler extract exhibited superior SOD-like activity compared to commercial herbal medicines. Therefore, it is determined that tomato juice containing heated extracted tomatoes is suitable for use in health functional foods, and it was confirmed that tomato juice extracted through a double-boiler process may be more suitable.

[0158] 2-3. Evaluation of XO Inhibitory Activity

[0159] The xanthine oxidase (XO) inhibitory activity of three tomato extracts with different solvents and extraction temperatures was evaluated according to the method of Stirpe and Corte (1969) as follows. 0.1 mL of a tomato extract sample diluted to a specific concentration was mixed with 0.6 mL of 0.1 M potassium phosphate buffer (pH 7.5) and 0.2 mL of a substrate solution containing 2 mM xanthine. 0.1 mL of xanthine oxidase (0.2 U / mL) was added to the mixture, and the reaction was stopped by adding 1 mL of 1 N HCl. The absorbance of the uric acid produced in the reaction solution was measured at 292 nm using a spectrophotometer. The XO inhibition rate was expressed as a percentage (%) of the difference in absorbance between the group with and without the tomato extract sample. As a control, L-ascorbic acid was measured using the same method as above and compared with each tomato extract.

[0160] Table 7 shows the results of evaluating the XO inhibitory activity according to the tomato extract concentrations of the Examples, Comparative Example 1, and Comparative Example 2.

[0161] Table 8 shows the results of measuring XO inhibitory activity according to the concentration of the control group, L-ascorbic acid (AsA).

[0162] Con.(%) Extract LRW (Comparative Example 1) LRJ (Comparative Example 2) LBW (Example) 1.25 - - - 2.5 5.20±0.96 5.38±2.62 - 5 5.31±1.47 b 7.64±1.31 a 8.06±1.46 a 10 7.64±2.23 c 10.59±0.80 b 18.77±1.61 a

[0163] AsA (ug / mL) 62.5 125 250 500 98.38±0.57 98.63±0.22 99.00±0.43 100.25±0.86

[0164] Figure 5 is a graph showing the results of evaluating the XO inhibitory activity according to the concentration of tomato extracts of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2.

[0165] Referring to Tables 7 and 8 and Figure 5, the XO inhibition rate at a 10% concentration was lower than that of L-ascorbic acid, a natural antioxidant, but the example, which was a water bath extract, showed an XO inhibition rate of 18.77%, which was about 2.4 times better than Comparative Example 1, which was a reflux extract.

[0166] Meanwhile, compared to the results showing that herbal medicines such as dried tangerine peel, green onion, and Achyranthes root exhibit an XO inhibition rate of less than 10% (Shin et al. (2013)), it can be seen that tomato decoction extract has a superior XO inhibition effect compared to dried tangerine peel, green onion, and Achyranthes root, which are herbal medicines also used as food.

[0167] Consequently, it was confirmed that the heated tomato extract has an XO inhibitory effect and is suitable for application in health functional beverages that can be used for the prevention and treatment of gout or inflammatory diseases; furthermore, it can be anticipated that the tomato extract obtained through a double-boiler extraction process may be more suitable.

[0169] 3. Evaluation of Anti-aging Activity

[0170] 3-1. Evaluation of Collagenase Inhibitory Activity

[0171] To measure the collagenase inhibitory effect, which is closely related to skin wrinkle formation, a modified method of Wunsch and Heindrich (1963) was used. 0.25 mL of a substrate solution containing 4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg (0.3 mg / mL) dissolved in 0.1 M Tris-HCl buffer (pH 7.5) in 4 mM CaCl2 was mixed with 0.1 mL of a sample solution diluted to a certain concentration. Then, 0.15 mL of collagenase (0.2 mg / mL) was added and the mixture was reacted at room temperature for 20 minutes. The reaction was stopped by adding 0.5 mL of 6% citric acid, and 1.5 mL of ethyl acetate was added and mixed. After standing for about 1 minute, only the supernatant was taken, and the absorbance was measured at 320 nm. Collagenase inhibition rate was expressed as a percentage (%) of the decrease in absorbance between the group with and without tomato extract samples, and as a control, EGCG (epigallocatechin gallate) extracted from green tea was measured using the same method as above and compared with tomato extracts with different extraction conditions.

[0172] Table 9 shows the results of evaluating the collagenase inhibition rate according to the tomato extract concentrations of the Examples, Comparative Example 1, and Comparative Example 2.

[0173] Table 10 shows the results of measuring the collagenase inhibition rate according to the concentration of the control group, EGCG.

[0174] Con.(%) Extract LRW (Comparative Example 1) LRJ (Comparative Example 2) LBW (Example) 1.25 - - 13.59±1.08 2.5 - - 29.76±1.85 5 - 4.76±1.14 32.23±1.73 10 - 8.21±2.98 36.05±1.61

[0175] EGCG(ug / mL) 62.5 125 250 500 47.26±4.47 66.67±1.69 67.93±2.11 77.36±3.64

[0176] Figure 6 is a graph showing the results of evaluating the collagenase inhibition rate according to the concentration of tomato extracts in one embodiment of the present invention, Comparative Example 1, and Comparative Example 2.

[0177] Referring to Tables 9, 10, and Figure 6, Example 1, which is a water bath extract, showed the best activity at 36.05% at a concentration of 10%, Example 2, which is a fresh juice extract, showed an effect of 8.21%, and Example 1, which is a reflux extract, showed no activity.

[0178] Meanwhile, when comparing the above results with the results of a study measuring the anti-wrinkle effects of some medicinal plants (Kim et al. (2008)), it was reported that water parsley showed a collagenase inhibition rate of 5.4% and Perilla frutescens 22.5%, indicating that the tomato decoction extract showed very excellent activity.

[0179] In conclusion, while further research is needed regarding the skin collagenase inhibitory effect of tomato extract, it was confirmed that it may be suitable for use in functional foods, including juice, for anti-aging and wrinkle improvement.

[0181] 3-2. Evaluation of Tyrosinase Inhibitory Activity

[0182] Whitening activity was expressed as the tyrosinase inhibition rate and measured according to the method of Yagi et al. (1987). 0.1 mL of tomato extract diluted to a specific concentration was mixed with 0.5 mL of 0.175 M sodium phosphate buffer (pH 6.8) and 0.2 mL of a substrate solution containing 10 mM L-DOPA. 0.2 mL of mushroom tyrosinase (110 U / mL) was added, and the mixture was reacted at 25°C for 2 minutes. The DOPA chrome produced in the reaction solution was measured at 475 nm using a spectrophotometer, and the tyrosinase inhibition activity was expressed as a percentage (%) of the absorbance reduction rate between the group with and without the tomato extract solution. Additionally, L-ascorbic acid, a natural antioxidant, was measured using the same method as a control and compared with the tyrosinase inhibition rates of each tomato extract.

[0183] Table 11 shows the results of evaluating the tyrosinase inhibitory activity according to the concentration of tomato extracts in the Examples, Comparative Example 1, and Comparative Example 2.

[0184] Table 12 shows the results of measuring tyrosinase inhibitory activity according to the concentration of the control group, L-ascorbic acid (AsA).

[0185] Con.(%) Extract LRW (Comparative Example 1) LRJ (Comparative Example 2) LBW (Example) 1.25 29.26±2.59a 14.04±2.74b 15.53±1.29b 2.5 27.54±2.59c 33.15±1.78b 49.84±2.91a 5 56.81±2.15c 60.24±1.34b 77.35±3.42a 10 69.48±0.94c 76.90±0.54b 86.52±5.14a

[0186] AsA (ug / mL) 62.5 125 250 500 96.21±2.01 96.34±3.88 97.02±1.17 98.10±0.85

[0187] Figure 7 is a graph showing the results of evaluating the tyrosinase inhibitory activity according to the concentration of tomato extracts of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2.

[0188] Referring to Tables 11 and 12 and Figure 7, it was found that at a concentration of 10%, the tyrosinase inhibitory activity was highest in the order of Example, Comparative Example 1, and Comparative Example 2, showing that the Example, which is a water bath extract, showed the best inhibitory effect, and also showed the best effect at concentrations of 2.5% and 5%.

[0189] Meanwhile, according to research results on the tyrosinase inhibitory effects of commercially available herbal medicines (Seo (2001)), it was reported that quince showed 20.5%, chili pepper 15.2%, Codonopsis pilosula 20.9%, Perilla frutescens 12.4%, and black pepper 33.2%. This indicates that the tomato decoction extract showed tyrosinase inhibitory activity approximately 2.6 to 5.6 times higher than that of the above herbal medicines, which are frequently used as herbal medicines, food, and spices.

[0190] In conclusion, tomato extract, which is utilized in food and health products, exhibits superior tyrosinase inhibitory activity compared to herbal medicines frequently used in food. Therefore, it is considered an excellent natural resource for use as a product additive to prevent browning or as a material for developing whitening products, and it is believed that it exhibits superior activity when heated compared to when not heated.

[0192] 4. Evaluation of cytotoxicity via MTT assay

[0193] 4-1. Cell Culture

[0194] The mouse macrophage RAW264.7 and human keratinocyte cell line HaCaT cells used in the experiment were cultured in dulbecco's modified eagle's medium (DMEM) containing 10% fetal bovine serum (FBS) and 1% antibiotic mytotic (10 units / mL penicillin, 100 µg / mL streptomycin) in an incubator supplied with 5% CO2 at 37°C. These cells were used for the MTT assay to measure the cytotoxicity of the tomato extract and for nitric oxide inhibition to measure anti-inflammatory activity.

[0196] 4-2. Cytotoxicity Assessment

[0197] The cytotoxicity of three tomato extracts obtained under different conditions was measured using the MTT [(3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazoliumbromide)] assay method with mouse-derived RAW264.7 macrophages and HaCaT keratinocytes. RAW264.7 macrophages were 1 × 10⁶ 4 It was diluted in DMEM medium to a concentration of 1 × 10⁶ cells / mL, and the keratinocyte HaCaT was 1 × 10⁶ 5 The extract was diluted in DMEM medium to a concentration of cell / mL, dispensed into 96-well plates (SPL Life Sci. Korea) at 200 μL aliquots, and incubated in a 5% CO2 incubator at 37°C for approximately 24 hours to allow adsorption onto the 96-well plates. Afterward, the supernatant was completely removed, and the tomato extract was diluted in FBS-free DMEM medium to prepare final concentrations of 12.5, 25, 50, 100, and 200 μg / mL. RAW264.7 macrophages and HaCaT cells were treated with each dilution at a concentration of 1 / 100 and cultured for 24 hours. Then, 100 µL of 5 µg / mL MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) solution was added to each well, and the cells were incubated in the dark at 37°C in a 5% CO2 incubator for 3 hours. After removing the supernatant, 100 µL of DMSO (dimethyl sulfoxide) was added to the purple formazen crystals formed by the reduction of MTT to lyse the cells. The cells were then stirred for approximately 10 minutes, and the absorbance was measured at 570 nm using an ELISA reader. Cytotoxicity was assessed by calculating the cell viability (% control) by comparing the groups with the tomato extract added with the control group without the extract, thereby determining the presence and extent of cytotoxicity.

[0198] Table 13 shows the results of evaluating cell viability for RAW264.7 according to the tomato extract concentrations of the Examples, Comparative Example 1, and Comparative Example 2.

[0199] Table 14 shows the results of evaluating the cell viability of HaCaT cells according to the tomato extract concentrations of the Examples, Comparative Example 1, and Comparative Example 2.

[0200] Extract Concentration (%) Non 1.25 2.5 5 10 LRW (Comparative Example 1) 100 102.41 105.26 105.19 107.64 LRJ (Comparative Example 2) 99.66 102.70 102.69 101.47 LBW (Example) 88.45 94.85 100.17 103.65

[0201] Extract Concentration (%) Non 1.25 2.5 5 10 LRW (Comparative Example 1) 100 10.524 107.55 107.13 103.56 LRJ (Comparative Example 2) 102.97 101.46 98.46 92.80 LBW (Example) 98.44 98.58 96.85 90.85

[0202] FIG. 8 is a graph showing the cell viability of RAW264.7 according to the tomato extract concentrations of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2.

[0203] FIG. 9 is a graph showing the cell viability of HaCaT cells according to the concentration of tomato extract in one embodiment of the present invention, Comparative Example 1, and Comparative Example 2.

[0204] Referring to Tables 13 and 14, Figures 8 and 9, for RAW264.7 macrophages, cell viability increased at a concentration of 10% in all extracts, and at a low concentration of 1.25%, the water-boiled extract of the example showed a cell inhibition rate of about 12%. All three extracts of the example, comparative example 1, and comparative example 2 showed a cell viability of 85% or higher.

[0205] As a result of confirming the cytotoxicity of tomato extract on HaCaT cells, which are skin keratinocytes, about 10% cytotoxicity was observed at a 10% concentration of LBW, but no toxicity that significantly reduced cell activity was observed in the Examples, Comparative Examples 1 and 2, so it is determined that it does not inhibit survival rate.

[0207] 5. Evaluation of Anti-inflammatory Effects

[0208] 5-1. Evaluation of NO Production Inhibitory Activity

[0209] To confirm the degree of immune enhancement by tomato extract, the concentration of NO produced in the culture supernatant of RAW264.7 macrophages was measured using a microplate assay. RAW264.7 macrophages were placed in a 6-well plate at a ratio of 1 × 10⁶ 5 Cells were seeded and cultured for 24 hours. Each extract was treated at different concentrations (25, 50, and 200 µg / mL), and lipopolysaccharide (LPS) at a concentration of 1 µg / mL was treated as a positive control and cultured for 24 hours. 100 µL of the supernatant from the cultured cells was mixed with an equal amount of Griess regent and reacted in a dark room at room temperature for 15 minutes; afterward, the absorbance was measured at 540 nm using an ELISA reader. The NO product inhibition rate was calculated by determining the difference between the absorbance values ​​of the experimental groups treated with each extract at different concentrations and the absorbance values ​​of the control group treated only with LPS.

[0210] Table 15 shows the results of evaluating the NO production inhibitory activity according to the concentration of tomato extracts in the Examples, Comparative Example 1, and Comparative Example 2.

[0211] Extract Concentration (%) Non 1.25 2.5 5 10 LRW (Comparative Example 1) 100 100.13 100.78 97.3 91.33 LRJ (Comparative Example 2) 101.80 99.74 98.13 93.14 LBW (Example) 101.47 96.74 91.66 88.81

[0212] Figure 10 is a graph showing the results of evaluating the NO production inhibitory activity according to the concentration of tomato extracts of one embodiment of the present invention, Comparative Example 1, and Comparative Example 2.

[0213] Figure 10 shows the results of measuring the amount of NO produced by treating RAW264.7 macrophages cultured in a 37°C 5% CO2 incubator with LPS, an endotoxin derived from Escherichia coli, to determine the NO production inhibitory activity of the three extracts of Examples and Comparative Examples 1 and 2.

[0214] Referring to Table 15 and Figure 10, the NO production rate decreased most significantly at 10% for the example with a water bath extract, by about 11%, while Comparative Example 1 with a reflux extract decreased by 9%, and Comparative Example 2 with a fresh juice extract decreased by about 7%, and the NO production rate decreased as the concentration increased.

[0215] NO is produced by immune-activated macrophages at the site of infection and plays an important role in neurotransmission, vasodilation, and immune modulation in various tissues. In conclusion, since the hot-boiled tomato extract inhibits NO production, it is considered that it can be utilized as a material for treating and preventing infectious responses and diseases. Furthermore, since the hot-boiled extract showed superior NO inhibitory activity compared to the fresh juice, it is expected that developing tomato products utilizing this could have an effect on suppressing inflammation.

[0217] 6. Statistical Processing

[0218] Experimental results measuring the beneficial components and anti-aging, antioxidant, and anti-inflammatory activities of tomato extracts extracted by different methods in Examples, Comparative Example 1, and Comparative Example 2 were independently repeated at least three times. Results were expressed as mean ± standard deviation using SPSS 27.0 for Windows program (Package for Social Science, SPSS Inc., Chicago IL, USA), and a one-way analysis of variance test (ANOVA est) was performed to test the statistical significance of the means between groups. Differences between groups were analyzed post-hoc using Duncan's multiple range test (p<0.05, a>b>c) at the p<0.05 level.

[0220] In this invention, tomatoes were extracted with or without heat application to verify their antioxidant, anti-aging, and anti-inflammatory effects according to the extraction method, with the aim of confirming the potential of tomatoes as materials for easily consumable products and cosmetic ingredients. As a result, it was confirmed that tomato extracts obtained by heat-based water bath extraction can be effective in terms of antioxidant, anti-aging, and anti-inflammatory activities compared to extracts obtained by other extraction methods.

[0222] Although the present invention has been described in detail through specific embodiments, this is for the purpose of specifically explaining the invention and is not limited thereto. It is evident that modifications or improvements can be made by those skilled in the art within the technical scope of the invention. All simple modifications or changes to the invention fall within the scope of the invention, and the specific scope of protection of the invention will be clarified by the appended claims.

Claims

Claim 1 Ii) a washing step for washing tomatoes; I-ii) a grinding step for grinding the tomatoes washed in step Ii); I-iii) a tomato extract preparation step comprising preparing a water bath extract by boiling the tomatoes ground in step I-ii) in water at a temperature of 90°C for 30 minutes and filtering the water bath extract; II-i) a first mixing step for preparing a mixture by adding salt, sugar, and purified water to the tomato extract of step I-iii); II-ii) a grinding step for grinding the mixture of step II-i); II-iii) a second mixing step for re-mixing the ground mixture of step II-ii); II-iv) a packaging step for packaging the re-mixed mixture of step II-iii) into a packaging container; II-v) a sterilization step for placing the packaged product of step II-iv) into a metal case and sterilizing it at a temperature of 80 to 90°C for 20 to 30 minutes; and II-vi) a step of drying the sterilized packaged product of step II-v); comprising, in step II-i), preparing a mixture by adding 0.1 parts by weight of salt, 0.6 parts by weight of sugar, and 4 parts by weight of purified water to 100 parts by weight of tomato extract, wherein the tomato extract is contained at a concentration of 10%, the content of polyphenols, flavonoid compounds, reducing sugars, and water-soluble proteins is increased, DPPH radical scavenging activity, SOD-like activity, XO inhibitory activity, collagenase inhibitory activity, tyrosinase inhibitory activity, and NO production inhibitory activity are increased, anti-aging activity and anti-inflammatory effect are improved, the reducing sugar content is increased, and the antioxidant activity is excellent. Claim 2 delete Claim 3 delete Claim 4 delete Claim 5 delete Claim 6 delete Claim 7 delete Claim 8 delete Claim 9 delete Claim 10 delete

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