Processed product of barley young leaf

Abstract

To provide a processed product of barley young leaves, which is safe when administered for a long time, can be used as a food or drink product and contains a high proportion of total polyphenols.SOLUTION: A processed product of barley young leaves has a total polyphenol content (catechin equivalent) of 1,350 mg or more per 100 g of dry mass, and / or an antioxidant ability of the barley young leaf extract measured by DPPH radical scavenging activity (50% scavenging concentration: IC50) of 3,000 μg / ml or less. Food and drink products containing the processed product of barley young leaves are expected to provide reliable effects based on the diverse physiological functions of the barley leaves due to its high total polyphenol content.SELECTED DRAWING: Figure 3

Description

[Technical field]

[0001] The present invention relates to a processed product of young barley leaves, and more particularly to a processed product of young barley leaves having a high content of total polyphenols. [Background technology]

[0002] Polyphenol is a general term for plant components that have two or more phenolic hydroxyl groups (hydroxyl groups bonded to aromatic rings such as benzene rings or naphthalene rings) in the molecule, and about 8,000 types have been identified so far. We ingest a wide variety of polyphenols on a daily basis from plant-based foods and beverages such as fruits, vegetables, and teas.

[0003] Years of research have shown that polyphenols are expected to have preventive or health-promoting effects against a variety of diseases, including obesity, cardiovascular disease, cancer, osteoporosis, neurodegenerative diseases, allergies, obesity, and muscle atrophy. Furthermore, a report states that "a meta-analysis of large-scale prospective cohort studies clearly shows that a high intake of dietary polyphenols reduces the risk of cardiometabolic diseases." The definitions, roles, functionality, future challenges and prospects of these polyphenols are described in detail in Non-Patent Documents 1 to 3.

[0004] It has long been known that some polyphenols contained in plant-based foods have strong antioxidant properties, and it has been thought that the preventive effects against various diseases and health promotion effects in humans and animals are based on this antioxidant ability. Furthermore, since the various effects cannot be explained by antioxidant activity alone, research on the bioavailability and target molecules of polyphenols is currently progressing (Non-Patent Document 1).

[0005] On the other hand, young wheat leaves, particularly young barley leaves, have been used safely for many years as plant-based food and drink, and have been reported to have a variety of physiological actions, such as antiulcer action, anticholesterol action, anti-inflammatory action, hypoglycemic action, antimutagenic action, antithrombotic action, vascular protection action, adipocyte differentiation inhibitory action, melamine synthesis inhibitory action, osteoporosis prevention action, and reactive oxygen scavenging enzyme (SOD) gene expression promotion action (see, for example, Non-Patent Document 4, and Patent Documents 1 to 4).

[0006] In addition, it has been reported that the ethyl acetate fraction of young barley leaves (Hordeum vulgare L.) has the effect of promoting ATP production and activating the Keap1-Nrf2 system, and that the active ingredients responsible for these actions include certain polyphenols (Patent Documents 8 and 9).

[0007] Furthermore, when young barley leaves are used as food or drink, various proposals have been made regarding their production methods (see, for example, Patent Documents 1 to 7).

[0008] Here, the amount of polyphenols contained in young barley leaves is approximately 1.333 g per 100 g of barley leaf powder (Non-Patent Document 5), and to the inventors' knowledge, barley leaves containing more polyphenols have not been reported. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] JP 2008-56580 A [Patent Document 2] JP 2011-51948 A [Patent Document 3] JP 2013-63940 A [Patent Document 4] JP 2015-140338 A [Patent Document 5] JP 2009-148213 A [Patent Document 6] JP 2003-33151 A [Patent Document 7] JP 2002-65204 A [Patent Document 8] Patent No. 6964290 [Patent Document 9] Patent No. 7201164 [Non-patent literature]

[0010] [Non-Patent Document 1] Yoshinori Nakamura, "Molecular mechanism of polyphenol functionality: How do polyphenols exert their functionality?" Journal of the Japanese Society for Food and Life Sciences, Vol. 33, No. 4, pp. 165-169 (2023) [Non-Patent Document 2] Junji Terao et al. "Prospects for functional research on dietary polyphenols and formulation of recommended intake amounts for social implementation of polyphenols" Chemistry and Biology Vol. 59, No.5, p.254-261, 2021 [Non-Patent Document 3] Kaeko Murota et al. "Introduction to polyphenol databases overseas and the current status and future challenges of polyphenol databases in Japan" Chemistry and Biology Vol. 59, No.9, p.46-472, 2021 [Non-Patent Document 4] Yoshihide Hagiwara et al., "Physiologically Functional Substances in Barley Leaf Green Juice Powder," Journal of the Japanese Society of Food Chemical Technology, Vol. 48, No. 10, pp. 712-725, October 2001 [Non-Patent Document 5] Nutritional information for young barley leaves (https: / / www.kanpo-yamamoto.com / oomugi / nutrition_facts.html) Summary of the Invention [Problem to be solved by the invention]

[0011] As mentioned above, young wheat leaves, especially young barley leaves, are substances with various and useful physiological effects that have been safely used for many years as food and drink, and it is presumed that many of these physiological effects (functionality) are due to the polyphenols they contain. Therefore, if young barley leaves with a high polyphenol content could be obtained, it would be considered to have better functionality.

[0012] That is, an object of the present invention is to provide a processed product of young barley leaves that is highly safe and has a high polyphenol content that can be taken safely over a long period of time, and to provide a food or drink containing the processed product of young barley leaves. [Means for solving the problem]

[0013] As a result of intensive research aimed at achieving the above object, the present inventors have found that young barley leaves grown in a certain environment contain high levels of total polyphenols, and that the amount of total polyphenols correlates with antioxidant activity. The present invention was achieved based on these findings.

[0014] That is, the gist of the present invention is as follows. (1) A processed product of young barley leaves, characterized by having either or both of the following properties (a) and (b): (a) The total polyphenol content (catechin equivalent) is 1,350 mg or more per 100 g of dry mass. (b) Antioxidant activity of barley leaf treatment products measured by DPPH radical scavenging activity (concentration showing 50% radical scavenging activity: IC 50 ) is less than 3,000 μg / ml. (2) An agent for improving antioxidant capacity containing a processed product of young barley leaves as an active ingredient, the antioxidant capacity of the processed product measured by DPPH radical scavenging activity (concentration showing 50% radical scavenging activity: IC 50 ) is 3,000 μg / ml or less. (3) A food or drink comprising the processed barley leaves described in (1) above or the antioxidant capacity enhancer described in (2) above. Effect of the Invention

[0015] According to the present invention, the processed barley leaves contain a high content of total polyphenols, and therefore, reliable effects based on the improved antioxidant ability and various physiological actions of the barley leaves described above can be expected. Furthermore, the processed barley leaves of the present invention have no safety issues and can be freely prepared in the form of food and drink, so that they can be taken over a long period of time not only by healthy people but also by the elderly, the sick, and those recovering from illness. [Brief description of the drawings]

[0016] [Figure 1] FIG. 2 is a diagram showing the experimental conditions for Example 3 (measurement of antioxidant capacity of young barley leaves). [Diagram 2] FIG. 2 is a diagram showing the experimental conditions for Example 3 (measurement of antioxidant capacity of young barley leaves). [Diagram 3] This figure shows the relationship between the total polyphenol content and antioxidant capacity of processed young barley leaves. In the figure, the vertical axis shows the total polyphenol content (mg / 100 g), and the horizontal axis shows antioxidant capacity [IC50 (μg / ml): concentration showing 50% scavenging activity of DPPH radicals)]. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] The following describes in detail an embodiment of the present invention, but the following description is merely an example of an embodiment of the present invention, and the present invention is not limited to the following description. Note that in this specification, when the expression "to" is used, it is used as an expression including the numerical values ​​or physical property values ​​before and after it.

[0018] In a preferred embodiment of the present invention, the young barley leaves have a high content of total polyphenols. As the young barley leaves, young barley leaves (stems and leaves) harvested from the start of tillering to the start of heading (when the plant height is about 20 to 40 cm) are preferably used.

[0019] Here, the barley may be any species and variety that belongs to the Hordeum vulgare species, such as two-row barley, four-row barley, six-row barley, and naked barley, but two-row barley, six-row barley, and naked barley are preferred, and six-row barley is particularly preferred.

[0020] More specifically, examples of varieties of two-rowed barley (malting barley) include Aominori, Agurimochi, Asaka Gold, Kinuyutaka, Oumiyutaka, Kirinijo, Haruna Nijo, Amagi Nijo, Fuji Nijo, Ishukushirazu, Misato Golden, Kawahonami, Kawamizuki, Kinuka Nijo, Komaki Nijo, Sakitama Nijo, Tone Nijo, Sachiho Golden, Satsukibare, Shunrei, and Mi These include Haru Gold, Sky Golden, Takachiho Golden, Tsuyushirazu, Tone Nijo, Nasu Nijo, Nishino Gold, Nishino Chikara, Nishino Hoshi, Haruka Nijo, Nirasaki Nijo, Nira Nijo, Harushizu, Satsuki Nijo, Houshun, Mikamo Golden, Miho Golden, Myogi Nijo, Yashio Golden, Yachiho Golden, Kitaiku 41, Irodori no Hoshi and Ryofu.

[0021] Examples of six-rowed barley varieties include Asamamugi, Kashimagor, Minorimugi, Katorimugi, Musashinomugi, Drillmugi, Sanadamugi, Haganemugi, Sayakaze, Suzukaze, Shunrai, Silky Snow, Shinjuboshi, Setugenmochi, Natori barley, Hamayutaka, Hayami barley, Fiber Snow, Baitori, and Miyuki barley.

[0022] Examples of naked barley varieties include Ichibanboshi, Senbonhadaka, Sanukihadaka, Kikaihadaka, Sanshu, Nanpuhadaka, Shiratamahadaka, Yunagihadaka, Daishimochi, Toyonokaze, Hayatehadaka, Akakamiriki, Ichiwase, Biwairohadaka, Benihadaka, Hashirihadaka, and Mannenboshi (formerly Mantenboshi), among others.

[0023] These varieties are cultivated by various agricultural experiment stations, agricultural research centers, private companies, etc., and their fruits and young leaves are used for food (including beverages). It is preferable to select and cultivate a variety suitable for the cultivation area from these varieties, and harvest the young leaves (stems and leaves).

[0024] The region for growing barley is not particularly limited, but a humid subarctic climate zone or a climate zone determined as a Köppen climate zone Dfb (cold zone, humid continental climate) is particularly preferred.

[0025] As described above, young barley leaves are harvested before the ear emergence stage, so there is no need to consider grain yield, and in terms of production efficiency, it is preferable to harvest young leaves sown in spring and summer. On the other hand, autumn sowing can ensure a longer growth period than spring sowing, so it may be more preferable for achieving a high content of total polyphenols and other components. On the other hand, barley has low cold resistance, so in highland and cold regions where winter survival is difficult, it is preferable to cultivate it in spring or summer.

[0026] In the present invention, the method for cultivating young barley leaves is not particularly limited as long as it can produce young barley leaves with a high total polyphenol content, but it is preferable to carry out spring cultivation in a subarctic humid climate region, which allows the production of young barley leaves with a high total polyphenol content.

[0027] The processed young barley leaves in the present invention may be in any form so long as they are processed young leaves (stems and leaves) that have grown to a height of about 20 to 40 cm from the start of tillering until the start of heading, as described above (hereinafter, this may be referred to as "processed young barley leaves").

[0028] Examples of processed young barley leaves include products that have been juiced, crushed, shredded, dried, roasted, extracted, powdered, or frozen, as well as products that have been juiced and dried into powder, dried, crushed, filtered (puree, juice), shredded, and extracted with hot water, and combinations of these products.

[0029] Among these, the most commonly used forms are a powdered product of the squeezed juice (squeezed juice) of young barley leaves (hereinafter sometimes referred to as "squeezed juice powder" or "extract powder") and a ground product of young barley leaves (hereinafter sometimes referred to as "ground powder").

[0030] Here, the squeezed juice powder can be prepared by harvesting (cutting) young barley leaves, squeezing the leaves to extract a liquid, mixing it with an appropriate excipient as necessary, and spray-drying or freeze-drying it in accordance with the methods described in Patent Documents 1 to 5, etc. This squeezed juice powder can be suitably used as the processed young barley leaves product of the present invention as it is, or by mixing it with other excipients, etc.

[0031] In addition, the ground powder can be prepared, for example, in accordance with the methods described in Patent Documents 6, 7, etc., by harvesting (cutting) young barley leaves, shredding them, drying and grinding them, and, if necessary, blanching them.

[0032] In the present invention, the processed young barley leaves have a high total polyphenol content. As described above, polyphenols having a wide variety of chemical structures are present as plant components. Therefore, the amount of polyphenols in the present invention is a value obtained by a method for measuring the "total amount" of polyphenols using Folin-Ciocalteu (phenol) reagent (hereinafter sometimes referred to as "Folin-Ciocalteu reagent"). This method has been adopted by the International Organization for Quantification as a method for quantifying the total polyphenols in green tea and black tea (ISO 14502-1:2005).

[0033] The Folin-Ciocalteu reagent contains phosphotungstic acid as an oxidizing agent, which reduces the phenolic hydroxyl group (forming tungsten molybdenum blue), giving it a blue color with a broad maximum absorption at 765 nm. Since the degree of coloring varies depending on the polyphenol component, in the present invention, catechin is used as a standard substance for the calibration curve to determine the total polyphenol amount (catechin equivalent). The measurement of the total polyphenol amount will be described in detail in the Examples.

[0034] In the present invention, the total polyphenol content (catechin equivalent) of the processed young barley leaves is usually 1,350 mg or more, preferably 1,400 mg or more, more preferably 1,500 mg or more, and even more preferably 1,600 mg or more per 100 g of dry mass of young barley leaves. There is no particular upper limit, and it may be the highest concentration that can be achieved by improving varieties and cultivation methods, but if one were to specify it, it would usually be 5,000 mg or less, preferably 3,000 mg or less.

[0035] In the present invention, the processed barley leaves have a high antioxidant capacity. The antioxidant capacity is determined by the concentration (IC 50 ) is evaluated.

[0036] DPPH is a stable radical that turns purple in solution, and changes to a non-radical form in the presence of a radical scavenger. By utilizing this property, the antioxidant capacity [IC 50 The antioxidant activity can be calculated by the following formula: (1) Concentration (mg / ml) exhibiting 50% radical scavenging activity. The measurement of antioxidant activity will be described in detail in the Examples.

[0037] In the present invention, the antioxidant capacity is defined as the concentration at which 50% of the DPPH radical is eliminated (IC 50), with the lower limit usually being 3,000 μg / ml or less, preferably 2,600 μg / ml or less, more preferably 2,300 μg / ml or less, and even more preferably 2,000 μg / ml or less. There is no particular restriction on the upper limit, which may be the highest concentration that can be achieved by improving varieties and cultivation methods, but if one were to specify it, it would usually be 600 μg / ml or more, preferably 1,000 μg / ml or more.

[0038] In the present invention, it is believed that by having the total polyphenol amount or antioxidant capacity within the above-mentioned range, the antioxidant capacity and various other functionalities possessed by young barley leaves can be more reliably expressed.

[0039] Thus, according to another aspect of the present invention, there is provided an agent for improving antioxidant capacity, which contains a processed product of young barley leaves as an active ingredient, and the antioxidant capacity of the processed product measured by DPPH radical scavenging activity (concentration showing 50% radical scavenging activity: IC 50 The present invention provides an antioxidant capacity enhancer having an antioxidant capacity of 3,000 μg / ml or less, and a food or drink comprising the antioxidant capacity enhancer.

[0040] The processed barley leaves of the present invention have high affinity with water, can be dissolved or suspended, and have excellent miscibility with ingredients that are usually added to foods, beverages, and medicines. Furthermore, barley leaves are an ingredient that has already been used for many years as foods and beverages, and are therefore highly safe. Therefore, they can be appropriately incorporated into daily dietary habits and ingested without difficulty and with peace of mind.

[0041] The processed barley leaves of the present invention are not particularly limited as long as they are in a form that can be orally ingested, such as a solution, suspension, emulsion, powder, solid molding, etc. Specific examples include powders, capsules, tablets, granules, powders, as well as forms used in ordinary foods and beverages such as beverages and foods, and pharmaceuticals. Suitable examples of the formulation form include powders, capsules, tablets, granules, drinks, etc., which allow easy adjustment of the intake amount.

[0042] In the present invention, the processed barley leaves may be used as they are, or may be used after preparing a desired preparation by a conventional method. For example, it may be mixed with various additives permitted for preparation and molded into a composition. The additives may be any additives that are added within a range that does not impair the effects of the present invention, and examples of such additives include herbal medicines, vitamins, minerals, excipients, surfactants, coating agents, oils and fats, stabilizers, preservatives, wetting agents, emulsifiers, lubricants, sweeteners, colorants, flavorings, buffers, antioxidants, pH adjusters, acidulants such as citric acid, malic acid, and gluconic acid, etc.

[0043] Examples of the excipients include lactose, starch, cellulose, maltitol, dextrin, etc. Examples of the surfactants include glycerin fatty acid esters, sucrose fatty acid esters, etc. Examples of the coating agents include gelatin, pullulan, shellac, zein, etc. Examples of the oils and fats include wheat germ oil, rice germ oil, safflower oil, etc. Examples of the waxes include beeswax, rice bran wax, carnauba wax, etc. Examples of the sweeteners include sucrose, glucose, fructose, stevia, saccharin, sucralose, etc.

[0044] Examples of the herbal medicines include Korean ginseng, American ginseng, Panax notoginseng, Ganoderma lucidum, propolis, agaricus, blueberries, ginkgo leaves and extracts thereof, etc. Examples of the vitamins include oil-soluble vitamins such as vitamin D and K, water-soluble vitamins such as vitamins B1, B2, B6, B12, C, niacin, pantothenic acid, folic acid, biotin, etc.

[0045] The present invention also provides a food or drink comprising a processed barley leaf product or an agent for improving antioxidant activity. Here, "comprises" means that the food or drink may contain a physiologically acceptable carrier depending on the desired product form, and may also contain other auxiliary ingredients that can be used in combination.

[0046] In the present invention, the term "food and drink" refers to any food and drink other than pharmaceuticals, and is not particularly limited as long as it is in a form that can be orally ingested as described above. Specific examples of such food and drink include instant foods such as instant noodles, retort foods, canned foods, microwave foods, instant soups, miso soups, freeze-dried foods, and other instant foods; beverages such as soft drinks, fruit juice drinks, vegetable drinks, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks, and alcoholic drinks; flour products such as bread, pasta, noodles, cake mixes, fried chicken powder, and bread crumbs; sweets such as candy, caramel, chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese sweets, and dessert sweets; and soybeans. These include condiments such as sauces, tomato processed seasonings, flavor seasonings, cooking mixes, sauces, dressings, soups, curry and stew bases; processed oils and fats, butter, margarine, mayonnaise, etc.; dairy products such as milk drinks, yogurt, lactic acid bacteria drinks, ice cream, cream, etc.; processed eggs, fish ham and sausages, fish paste products, etc.; processed livestock products such as meat ham and sausages; processed agricultural products such as canned agricultural goods, jams, marmalades, pickles, boiled beans, cereals, etc.; frozen foods, etc.

[0047] Furthermore, the food and beverage categories include health foods (e.g., functional foods, nutritional supplements, health supplements, nutritionally fortified foods, nutritionally regulated foods, supplements, etc.), health functional foods (e.g., foods for specified health uses, nutritionally functional foods, foods with functional claims, etc.), foods for special dietary uses (e.g., foods for the sick, infant formula, milk powder for pregnant or breastfeeding women, etc.), as well as foods and beverages labeled for risk reduction, prevention, or improvement of diseases or conditions (symptoms) caused by decreased functions that are expected to be improved by taking young barley leaves.

[0048] The intake amount of the processed barley leaves, antioxidant capacity improving agent, food and drink of the present invention is difficult to uniformly define due to the type and form of the preparation, the purpose of prevention or improvement, etc., but for example, for an adult (body weight 60 kg), it is usually about 15 mg to 15 g, preferably about 50 mg to 9 g, per day as squeezed barley leaves powder, and for example, it is usually about 50 mg to 15 g, preferably about 100 mg to 9 g, per day as crushed barley leaves powder. Furthermore, the content in the preparation may be appropriately set in consideration of the daily intake so that the required daily intake can be taken. EXAMPLES

[0049] The present invention will be described in more detail below with reference to examples (experimental examples), but the technical scope of the present invention is not limited to these examples.

[0050] [Example 1] Production of barley leaf juice powder Young leaves (stems) of barley cultivated in spring in a subarctic humid climate zone were squeezed in accordance with the method described in JP 2009-148213 A (Patent Document 5), and 5 kg of juice was obtained by freeze-drying to obtain 223 g of juice powder.

[0051] [Example 2] Total polyphenol content in barley leaf juice powder The total amount of polyphenols contained in the young barley leaf juice powder obtained in Example 1 (hereinafter sometimes referred to as "Example 1 extract powder") was measured using a Folin-Ciocalteu phenol reagent as follows.

[0052] The reagents and instruments used for the measurements are as follows: Folin-Ciocalteu phenol reagent; manufactured by SIGMA-ALDRICH, Inc. Sodium carbonate (Na2CO3); Kanto Chemical Co., Ltd. CAS: 497-19-8 FW: 105.99 ·(+)-Catechin hydrate; manufactured by Cayman Chemical Inc. CAS: 154-23-4 FW:290.27 Ethanol (EtOH); Kishida Chemical Co., Ltd. CAS: 140-28553 FW: 46.07 Spectrophotometer: Shimadzu UVmini-1240 Electronic balance: GR-60 made by A&D Co., Ltd.

[0053] First, 10 mg of catechin hydrate for preparing the calibration curve was weighed on an electronic balance and placed in a 10 ml measuring flask, and 8 ml of 50% ethanol solution (hereinafter abbreviated as "50% EtOH") was added and stirred. After stirring, the solution was made up to 62.5 μg / ml, 125 μg / ml, 250 μg / ml, 500 μg / ml, and 1,000 μg / ml with 50% EtOH to prepare five samples for the calibration curve (hereinafter sometimes referred to as "Solution A").

[0054] Next, 100 mg of the extract powder of Example 1 was weighed using an electronic balance, placed in a 10 ml measuring flask, 8 ml of 50% EtOH was added, stirred, and left to stand for 1 hour. After 1 hour, the solution was diluted with 50% EtOH (10 mg / ml). After filtering through a 0.45 μm filter, the solution was diluted to 1 mg / ml and 5 mg / ml to prepare three measurement samples (hereinafter sometimes referred to as "Solution B").

[0055] 6 ml of Folin-Ciocalteu phenol reagent was diluted with 24 ml of pure water (5-fold diluted phenol reagent). 3 g of sodium carbonate (Na2CO3) was weighed on an electronic balance and dissolved in 30 ml of pure water (hereinafter referred to as "Solution C").

[0056] 50 μl each of solution A (catechin hydrate: 5 samples) and solution B (extract powder of Example 1: 3 samples) were added with 4 ml of pure water, 1 ml of 5-fold diluted phenol reagent, and 1 ml of solution C, and the mixture was allowed to react in the dark for 1 hour.

[0057] After one hour, colorimetric quantification (Abs 760 nm) was performed, and the total polyphenol amount (catechin equivalent) of the extract powder of Example 1 was calculated from a calibration curve prepared from the measurement results of solution A (catechin hydrate).

[0058] As a result, the total polyphenol content (catechin equivalent) of the extract powder of Example 1 was 1,624 mg / 100 g at an added concentration of 5 mg / ml, and 1,682 mg / 100 g at an added concentration of 10 mg / ml. Note that the absorbance of the extract powder of Example 1 at an added concentration of 1 mg / ml was excluded because it was outside the range of the calibration curve.

[0059] [Example 3] Measurement of antioxidant capacity of young barley leaves The antioxidant capacity of young barley leaves grown in different regions was measured.

[0060] The climate of the growing region of the young barley leaves used and its Köppen classification (Köppen climate classification) are as follows: Example 1 Extract Powder: Barley leaf juice powder prepared in Example 1 Subarctic humid climate Köppen classification Dfb (cold, humid continental climate) · Young barley leaves A (ground powder from China) China; Yunnan Province Temperate, subtropical, tropical Köppen classification (Overall, warm winter low rainfall climate (Köppen classification Cwb / Cwa), warm humid climate (Cfa) near the border with Vietnam in the southeast, cool winter low rainfall climate (Dwb / Dwc) in high altitude areas in the northwest) · Young Barley Grass B (American extract powder) Native to the United States; Utah, continental dry climate, Köppen classification Dsa (subarctic (cold), highland Mediterranean climate) · Young barley leaves C (crushed powder from Kyushu) Kyushu region; temperate Seto Inland Sea climate; Köppen classification Cfa (warm and humid climate)

[0061] The antioxidant capacity of the above young barley leaves was evaluated by measuring the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity as follows.

[0062] The reagents and instruments used for the measurements are as follows: ·MES sodium salt; Manufactured by SIGMA-ALDRICH, Inc. CAS:71119-23-8 FW:217.2 HCl (hydrochloric acid); SIGMA-ALDRICH, Inc. CAS: 7647-01-0 FW: 36.46 ·DPPH (1,1-diphenyl-2-picrylhydrazyl) free radical; TOKYO CHEMICAL INDUSTRY CO.,LTD. CAS: 1898-66-4 FW: 394.32 Ethanol (EtOH); Kishida Chemical Co., Ltd. CAS: 140-28553 FW: 46.07 ·EGCG (epigallocatechin gallate); Wako Pure Chemical Industries, Ltd. CAS: 059-05411 FW: 458.37 Microplate reader: Thermo Fisher Scientific VARIOSKAN FLASH ·Electronic balance; made by METTLER TOLEDO AG104

[0063] In a fume hood, 10 ml of HCl (hydrochloric acid) was taken using a safety pipetter and measuring pipette, placed in a 200 ml glass measuring cylinder that had already contained 100 ml of Milli-Q water, and the solution was made up to 120 ml with Milli-Q water to make 1 M HCl.

[0064] 2.172 g of MES sodium salt was weighed out using an electronic balance and transferred to a 100 ml beaker. 50 ml of Milli-Q water was added to the beaker and stirred with a stirrer. The pH meter was calibrated at three points, and 1 M HCl was added until the pH was 6.1. The solution was then diluted to 100 ml with Milli-Q, placed in a glass bottle, and stored at 4°C until use (hereinafter referred to as "MES solution").

[0065] 11.7 mg of DPPH was weighed out on an electronic balance and placed in a 50 ml measuring flask. 40 ml of ethanol was added, and the flask was covered with aluminum foil to block light, and then ultrasonicated for 30 minutes, and the volume was increased to 50 ml. The solution was placed in a glass bottle, protected from light with aluminum foil, and stored at 4°C until use (hereinafter referred to as "DPPH solution").

[0066] 250 ml of ethanol was placed in a 500 ml graduated cylinder and mixed well with 250 ml of Milli-Q water placed in another 500 ml graduated cylinder to prepare a 50% aqueous ethanol solution (hereafter abbreviated as "50% EtOH"). The solution was placed in a medium bottle and stored at room temperature.

[0067] 5 mg of EGCG (epigallocatechin gallate) was weighed out on an electronic balance and placed in a 15 ml centrifuge tube, and 5 ml of 50% EtOH was added and stirred. 1 ml of it was pipetted into a new 15 ml centrifuge tube, and 9 ml of 50% EtOH was added to dilute it 10 times. The same procedure was repeated to prepare a 100-fold diluted solution, which was used as a positive control (hereinafter sometimes referred to as "positive control").

[0068] 4 ml of DPPH solution, 1 ml of MES solution, and 3 ml of Milli-Q water were placed in a 15 ml centrifuge tube, stirred, and shielded from light with aluminum foil (Solution 1).

[0069] 4 ml of 50% EtOH, 1 ml of MES solution, and 3 ml of Milli-Q water were placed in a 15 ml centrifuge tube and stirred (Solution 2).

[0070] Example 1 Extract powder was weighed out by an electronic balance (100 mg), placed in a 15 ml centrifuge tube, and 9.9 ml of 50% EtOH was added, stirred for 1 minute, and allowed to stand for 60 minutes. It was then filtered through a 0.45 μm filter (PVDF) and placed in a new 15 ml centrifuge tube (10,000 μg / ml). This was diluted with 50% EtOH to 1,000 μg / ml, 1,500 μg / ml, and 2,000 μg / ml.

[0071] 100 mg of young barley leaves A was weighed on an electronic balance and placed in a 15 ml centrifuge tube, 9.9 ml of 50% EtOH was added, stirred for 1 minute, and left to stand for 60 minutes. It was then filtered through a 0.45 μm filter (PVDF) and placed in a new 15 ml centrifuge tube (10,000 μg / ml). This was then diluted with 50% EtOH to 2,000 μg / ml, 3,000 μg / ml, and 4,000 μg / ml.

[0072] 100 mg of young barley leaves B was weighed on an electronic balance and placed in a 15 ml centrifuge tube, 9.9 ml of 50% EtOH was added, stirred for 1 minute, and left to stand for 60 minutes. It was then filtered through a 0.45 μm filter (PVDF) and placed in a new 15 ml centrifuge tube (10,000 μg / ml). This was then diluted with 50% EtOH to 3,000 μg / ml, 4,000 μg / ml, and 5,000 μg / ml.

[0073] 100 mg of young barley leaves C was weighed on an electronic balance and placed in a 15 ml centrifuge tube, 9.9 ml of 50% EtOH was added, stirred for 1 minute, and left to stand for 60 minutes. It was then filtered through a 0.45 μm filter (PVDF) and placed in a new 15 ml centrifuge tube (10,000 μg / ml). This was then diluted with 50% EtOH to 5,000 μg / ml, 6,000 μg / ml, and 7,000 μg / ml.

[0074] 100μl of 50% EtOH was added as a blank to B2-B5 of the 96-well plate, 100μl of positive control was added to C2-C5, 1,000μg / ml of the extract powder of Example 1 was added to B7-B10, 1,500μg / ml to C7-C10, 2,000μg / ml to D7-D10, 2,000μg / ml of young barley leaves A was added to E7-E10, 3,000μg / ml to F7-F10, and 4,000μg / ml to G7-G10. Next, 150μl of solution 2 was added to B2, C2, B7, C7, D7, E7, F7, and G7, and pipetted 10 times. 150 μl of solution 1 was added to B3-B5, C3-C5, B8-B10, C8-C10, D8-10, E8-E10, F8-F10, and G8-G10, and pipetted 10 times (Figure 1). After leaving the plates in the dark for 20 minutes, the absorbance at 550 nm was measured using a plate reader.

[0075] 100μl of 50% EtOH was added as a blank to B2-B5 of the 96-well plate, 100μl of positive control was added to C2-C5, 3,000μg / ml of young barley leaves B was added to B7-B10, 4,000μg / ml to C7-C10, 5,000μg / ml to D7-D10, 5,000μg / ml of young barley leaves C was added to E7-E10, 6,000μg / ml to F7-F10, and 7,000μg / ml to G7-G10. Next, 150μl of solution 2 was added to B2, C2, B7, C7, D7, E7, F7, and G7, and pipetted 10 times. 150 μl of solution 1 was added to B3-B5, C3-C5, B8-B10, C8-C10, D8-10, E8-E10, F8-F10, and G8-G10, and pipetted 10 times (Figure 2). After leaving the plates in the dark for 20 minutes, the absorbance at 550 nm was measured using a plate reader.

[0076] First, the antioxidant rate was calculated using the following formula.

[0077] Antioxidant rate (%) = (1-mean absorbance value of each barley leaf ÷ mean value of blank) x 100

[0078] Next, a calibration curve was created from the obtained antioxidant rate, and the antioxidant capacity (IC 50 ) was calculated.

[0079] As a result, the antioxidant activity (IC 50 ) was 1,876.9 μg / ml, barley leaf A was 3,447.5 μg / ml, barley leaf B was 4,400.9 μg / ml, and barley leaf C was 6,667.5 μg / ml.

[0080] [Example 4] Measurement of total polyphenols in young barley leaves The total polyphenol content was measured in young barley leaves grown in different regions.

[0081] The origin of the young barley leaves used and the Köppen classification (Köppen climate classification) of that region are as shown in Example 3.

[0082] 10 mg of catechin hydrate for preparing the calibration curve was weighed out on an electronic balance and placed in a 10 ml measuring flask, and 8 ml of 50% ethanol solution (hereinafter abbreviated as "50% EtOH") was added and stirred. After stirring, the solution was made up to 62.5 μg / ml, 125 μg / ml, 250 μg / ml, 500 μg / ml, and 1,000 μg / ml with 50% EtOH to prepare five samples for the calibration curve (hereinafter sometimes referred to as "Solution A").

[0083] 200 mg of young barley leaves A was weighed out on an electronic balance and placed in a 10 ml measuring flask, 8 ml of 50% EtOH was added, stirred, and left to stand for 1 hour. After 1 hour, the solution was made up to 50% EtOH and filtered through a 0.45 μm filter (hereinafter referred to as "young barley leaves A solution").

[0084] 300 mg of young barley leaves B was weighed on an electronic balance and placed in a 10 ml measuring flask, 8 ml of 50% EtOH was added, stirred, and left to stand for 1 hour. After 1 hour, the solution was made up to 50% EtOH and filtered through a 0.45 μm filter (hereinafter referred to as "young barley leaves B solution").

[0085] 400 mg of young barley leaves C was weighed out on an electronic balance and placed in a 10 ml measuring flask, 8 ml of 50% EtOH was added, stirred, and left to stand for 1 hour. After 1 hour, the solution was made up to 50% EtOH and filtered through a 0.45 μm filter (hereinafter referred to as "young barley leaves C solution").

[0086] 6 ml of Folin-Ciocalteu phenol reagent was diluted with 24 ml of pure water (5-fold diluted phenol reagent). 3 g of sodium carbonate (Na2CO3) was weighed on an electronic balance and dissolved in 30 ml of pure water (hereinafter referred to as "Solution C").

[0087] To 50 μl each of solution A (catechin hydrate: 5 samples), young barley leaf A solution, young barley leaf B solution, and young barley leaf C solution, 4 ml of pure water, 1 ml of 5-fold diluted phenol reagent, and 1 ml of solution C were added and reacted in the dark for 1 hour.

[0088] After one hour, colorimetric quantification (Abs 760 nm) was performed, and the total polyphenol amount (catechin equivalent) of each barley leaf was calculated from a calibration curve created from the measurement results of solution A (catechin hydrate).

[0089] As a result, the total polyphenol content (catechin equivalent) of barley leaf A was 1,224.4 mg / 100 g, that of barley leaf B was 739.2 mg / 100 g, and that of barley leaf C was 691.9 mg / 100 g.

[0090] [Example 5] Comparative study of total polyphenols and antioxidant capacity of young barley leaves

[0091] Using the results obtained in Examples 2, 3, and 4, a comparative study was conducted on the total polyphenols and antioxidant capacity of young barley leaves grown in different regions. The results are shown in Figure 3. As can be seen from the figure, there is a correlation between the total polyphenol content and antioxidant capacity of young barley leaves, and the extract powder (Extract powder of Example 1) of young barley leaves grown in a subarctic humid climate (Köppen classification Dfb) has a high total polyphenol content and high antioxidant capacity (IC 50 (The concentration at which 50% of the radicals are eliminated is the smallest.)

Claims

1. 1. A processed product of young barley leaves, characterized by having either or both of the following properties (a) and (b): (a) The total polyphenol content (catechin equivalent) is 1,350 mg or more per 100 g of dry mass. (b) Antioxidant activity of the treated barley leaves measured by DPPH radical scavenging activity (concentration showing 50% radical scavenging activity: IC ５０ ) is 3,000 μg / ml or less.

2. An agent for improving antioxidant capacity containing a processed product of young barley leaves as an active ingredient, the antioxidant capacity of the processed product measured by DPPH radical scavenging activity (concentration showing 50% radical scavenging activity: IC ５０ ) is 3,000 μg / ml or less.

3. 10. A food or drink comprising the processed young barley leaves according to claim 1 or the antioxidant activity enhancer according to claim 2.

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