WO2007110924A1

WO2007110924A1 - Method of oxidation inhibition using volatile component

Info

Publication number: WO2007110924A1
Application number: PCT/JP2006/306264
Authority: WO
Inventors: Teruo Miyazawa; Kiyotaka Nakagawa; Jun Kariya
Original assignee: Tohoku University; Project M Co., Ltd.
Priority date: 2006-03-28
Filing date: 2006-03-28
Publication date: 2007-10-04
Also published as: JPWO2007110924A1

Abstract

A method of oxidation inhibition that excels in antioxidative potency, being safe and ensures high versatility. There is provided a method of oxidation inhibition, comprising causing a phenolic compound in volatile form to coexist with an object substance.

Description

Antioxidation method using volatile components

Technical field

[0001] The present invention evaluates the oxidation prevention method using volatilized phenolic compounds, the antioxidant containing the phenolic compounds, and the antioxidant ability of the test substance in the volatile state. On how to do.

Background art

[0002] When fats and oils and fat foods are stored for a long time, they are oxidized by oxygen in the air, and the color tone and flavor are impaired. Vegetable oils and fish oils rich in unsaturated fatty acids are particularly susceptible to oxidation. Oils and fats produce peracid lipids, and their overdose is a concern for toxicity. Therefore, a method for preventing acidification of foods, particularly foods containing oils and fats, is important.

[0003] As a method for preventing oxidation of food, at present, an anti-oxidation agent is frequently used. Antioxidants include natural antioxidants such as ascorbic acid (vitamin C), tocopherol (vitamin E) and flavonoids, and synthetic antioxidants such as butylhydroxylazole (BHA) and dibutylhydroxytoluene (BHT). It has been known. However, ascorbic acid is a water-soluble compound and is difficult to use in oil and fat foods. Tocopherol needs to be used in combination with other anti-oxidant agents that have a very high anti-acid power. Flavonoids are colored and have limited use. Synthetic acid inhibitors, such as BHA and BHT, are a safety issue and their use in foods and pharmaceuticals is restricted. Against this background, the development of excellent antioxidants to replace conventional antioxidants is desired!

[0004] In addition, a method has been reported in which 1,8-cineole, which has anti-acidic action in the air, is volatilized and suspended in the human body (Y. Saito et al., Biosci. Biotechnol. Biochem., 68, 781-786 (2004), JP 2005-110760). However, the strong and volatilized 1,8-cineole does not have enough anti-acidic ability for the living body.

Disclosure of the invention

[0005] An object of the present invention is to provide a method for preventing oxidation that is excellent in antioxidative power, safe and versatile. [0006] As a result of intensive studies to achieve the above-mentioned object, the present inventors have found that an excellent anti-oxidative effect is exhibited by using a phenolic compound in a volatilizable state. The present invention has been completed.

That is, the present invention includes the following inventions.

[0008] (1) A method for preventing acidification, which comprises allowing a phenolic compound to coexist with a target substance in a volatilizable state.

[0009] (2) A phenolic compound is represented by the following formula I:

[Chemical 1]

[0010] [where

R ¹ may have a substituent, may be C alkyl or have a substituent! /, May! /, C

1-6]

-6

R ² may have a hydrogen atom or a substituent, or may have C alkyl or a substituent! /

1-6

Yo! /, C alkenyl, substituted! /, May! /, C alkynyl or substituted

2-6 2-6

/! Or even! /, C alkoxy,

1-6

R ^{3 to} R ⁵ are each independently a hydrogen atom or a substituent]

The antioxidant method according to (1), which is a compound represented by the formula:

[0011] (3) The antioxidant method according to (1) or (2), wherein the target substance is an oily component or a substance containing an oily component.

[0012] (4) The phenolic compound is at least one selected from the group forces of guaiacol, bulguaiacol, eugenol, isoeugenol and 2,4,6-trimethylphenol (1) to (3 ) The method for preventing acidification according to any of the above.

[0013] (5) The following formula I: [Chemical 2]

[0014] [where

R ¹ may have a substituent, may be C alkyl or have a substituent! /, May! /, C

1-6]

-6

1-6

Yo! /, C alkenyl, substituted! /, May! /, C alkynyl or substituted

2-6 2-6

/! Or even! /, C alkoxy,

1-6

R ^{3 to} R ⁵ are each independently a hydrogen atom or a substituent]

The antioxidant containing the phenolic compound represented by these.

[0015] (6) The antioxidation agent according to (5) for preventing oxidation of an oily component or a substance containing an oily component.

[0016] (7) The antioxidant according to (5) or (6), comprising at least one selected from the group forces of guaiacol, bulguaiacol, eugenol, isoeugenol and 2,4,6 trimethylphenol.

[0017] (8) The antioxidant according to any one of (5) to (7), wherein the phenolic compound is in a volatizable state.

[9] (9) A method for evaluating the antioxidant capacity of a test substance in a volatile state,

a) measuring the absorbance of a solution containing linoleic acid and β-strength,

b) separately placing the solution and the test substance in a sealed container;

c) measuring the absorbance of a solution containing linoleic acid and j8-strengthen after a certain period of time; and

d) calculating the difference between the absorbance measured in step a and the absorbance measured in step c; Said method comprising

(10) A method for evaluating the antioxidant ability of a test substance against an oily component in a volatile state.

a) measuring the absorbance of the oily component to which linoleic acid and β-carotene are added, b) placing the oily component and the test substance separately in a sealed container,

c) a step of measuring the absorbance of the oily component after a certain period of time; and

d) calculating the difference between the absorbance measured in step a and the absorbance measured in step c.

[0019] (11) A method for evaluating the antioxidant ability of a test substance against an oily component in a volatile state,

a) measuring the amount of malondialdehyde in the oil component;

b) separately placing the oily component and the test substance in a sealed container;

c) a step of measuring the amount of malondialdehyde in the oil component after a certain period of time, and

d) calculating the difference between the amount of malondialdehyde measured in step a and the amount of malondialdehyde measured in step c;

Including said method.

Brief Description of Drawings

FIG. 1 shows an embodiment of the evaluation method of the present invention.

[FIG. 2] FIG. 2 shows the amount of fading of | 8-carotene after 150 minutes when various substances are added.

[FIG. 3] FIG. 3 shows the amount of fading of the / 3 straight mouth ten days after the addition of various substances.

FIG. 4 shows changes in malondialdehyde after 5 days when various substances are added. BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, the present invention will be described in detail.

[0022] In one embodiment, the present invention relates to a method for preventing acidification, which comprises causing a phenolic compound to coexist with a target substance in a volatizable state.

[0023] In this specification, the phenolic compound refers to a compound having a skeleton structure in which 1 to 3 hydroxyl groups are directly bonded to a benzene ring, and the benzene ring further having a substituent. Book In the present invention, phenolic compounds do not include phenol itself! /. Here, the substituents are halogen, C alkyl, C alkyl, C alkyl, C alkoxy.

1-6 2-6 2-6 1-6 cis, amino-containing C-anolenoquinamino, di-C-anolenoquinamino, cyano-containing isocyanate, nitro

1-6 1-6

, Nitroso, carboxyl, mercapto, C alkylthio, etc., preferably

1-6

Halogen, c alkyl, C alkyl, C alkyl, C alkoxy

1-6 2-6 2-6 1-6

The The number of substituents is preferably 1 to 3.

In this specification, halogen is preferably selected from fluorine, chlorine, bromine or iodine.

[0025] In the present specification, C alkyl, C alkyl, C alkyl, C alkyl

1-6 2-6 2-6 1-6 Coxy, alone or as part of another group, includes alkyl, alkyl, alkyl, or alkoxy containing the specified number of carbon atoms, respectively. It may be linear or branched.

[0026] The phenolic compound of the present invention preferably has one hydroxyl group directly bonded to the benzene ring, and in this case, C alkoxy or C

1-6 1 Has alkyl.

-6

In the present invention, a compound having volatility at room temperature, that is, about 25 ° C. is used as the phenolic compound.

[0028] The phenolic compound is preferably of formula I:

[Chemical 3]

[0029] [where

R ¹ may have a substituent, may be C alkyl or have a substituent! /, May! /, C

1-6

Anoroxixi, R ² may have a hydrogen atom or a substituent, or may have C alkyl or a substituent! /

1-6

Yo! /, C alkenyl, substituted! /, May! /, C alkynyl or substituted

2-6 2-6

/! Or even! /, C alkoxy,

1-6

R ^{3 to} R ⁵ are each independently a hydrogen atom or a substituent]

The compound represented by these is mentioned.

[0030] In the present invention, the phenolic compound containing the compound of the formula I includes stereoisomers such as several isomers, rotational isomers and optical isomers of the above compounds, and salts and solvates. Is included. The salt is not particularly limited, and examples thereof include ammonium salts such as trimethylammonium salt, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, and the like. Is mentioned.

[0031]! Examples of the substituent in ^ ˜ include, for example, halogen, C alkyl, C alk-

1-6 2-6, C alkyl, C alkoxy, amide C alkyl amide di C alkyl

2-6 1-6 1-6 1-6 Cyanated isocyanate, nitro, nitroso, carboxyl, mercapto, C-al

1-6 Kirthio and the like. In R ¹ and R ² , C alkyl, C alkenyl,

1 -6 2-6

C alkyl and C alkoxy have a maximum of three substituents.

2-6 1 -6

[0032] R ¹ is preferably C alkyl or C alkoxy, more preferably C alkyl.

1-6 1 -6 1-3 or C alkoxy, especially methoxy or methyl.

13

[0033] R ² is preferably a hydrogen atom, C alkyl, C alkyl or C alkyl.

1-6 2-6 2-6.

[0034] R ^{3 to} R ⁵ are preferably each independently a hydrogen atom, halogen or C alkyl.

1-6.

[0035] When R ¹ is C alkyl, preferably R ² and R ³ are each independently C

1-6 1-6 alkyl, and R ⁴ and R ⁵ are hydrogen atoms.

[0036] When R ¹ is C alkoxy, preferably R ² is C alkell and R ^{3 to} R ⁵

1-6 2-6

Are all hydrogen atoms.

[0037] Specific examples of the phenolic compound in the present invention include 4-aryl 2,6-dimethoxyphenol, 4-arylphenol, force rubachlor, catechol, cresol, m-taresole, o-cresol, p-cresol, dihydro Eugenol, 2, 6 Dimethoki Siphenol, 2,3-Dimethylphenol, 2,6 Dimethylphenol, 3,5 Dimethylphenol, p-Ethoxyphenol, 4-Ethylda Ayacol, 2 Ethenophenol, 3 Ethylphenol, 4 Ethylphenol, 2 Ethylthioenol, Eugen Nonole, Guayako Nore, Vininole Guayako Nore, Hexenoresi Hexinorecetanol, 4-Hydroxybenzoitquaside, 4-Hydroxybenzylethyl ether, Isoeugenol, 2-Isopropylphenol, 4-Isopropylphenol, 3- Methoxy-1-5-methylphenol, 3-methoxyphenol, 4-methoxyphenol, 4-methyl-2,6-dimethoxyphenol, 3,4-methylenedioxyphenol, 5-methyldiaacol, 4 (methylthio) phenol p-propylphenol, protocatechuic acid, resorcinol, salicylic acid, syringeic acid, thymol, thioguaiacol, nitropropyleneglycolanolasetal, nonitrop, vinyloreguar alcohol, p vinylol phenol, 2,4 xylenol, 2,5 xylenol, 3,4 xylenol, 2, 3, 6-trimethylphenol, o-propylphenol, 3-tert-butylphenol, etc., guaiacol, burguaiacol, eugenol, isoeugenol and 2 3, 6 Trimethylphenol is preferred. These structures are shown below.

[Chemical 4]

Isogyenol — 卜 Lime Enol [0038] The phenolic compound may be used alone or in combination of two or more.

[0039] The present invention is characterized in that the phenolic compound as described above coexists with the target substance in a volatizable state.

[0040] The phenolic compound may be a synthetic compound or a natural one extracted from a plant. For example, guaiacol can extract forces such as sasa, eugenol can extract forces such as clove, keihi, -kuzuku, pimento, and isoeugenol can extract forces such as kuzukuku. Furthermore, it is possible to use a phenolic compound produced under the processing conditions of food and natural products. For example, vinyl diaiacol produced during roasting of coffee can be mentioned. These phenolic compounds have the advantages of excellent antioxidant ability, safety and high versatility.

[0041] To coexist with a target substance in a volatilizable state means to allow the phenolic compound to coexist with the target substance in a volatilized state, or to preserve the phenolic property for at least a certain period in the process of storage or the like. It means that the compound is volatilized and coexists with the target substance. By allowing the phenolic compound to coexist with the target substance in a volatilized state, the phenolic compound that has been vaporized into gas comes into contact with the target substance and exhibits an antioxidant effect. In other words, the method includes volatilizing the phenolic compound and bringing it into contact with the target substance, or bringing the phenolic compound in a gaseous state into contact with the target substance.

[0042] As long as the phenolic compound can be volatilized and the volatilized phenolic compound and the target substance are in contact with each other, the target substance and the phenolic compound exist separately. Let me! You can mix phenolic compounds with the target substance!

[0043] By allowing the target substance and the phenolic compound to coexist in a somewhat closed space, preferably in a sealed space, the concentration of the volatile phenolic compound in the space can be reduced. In addition, the volatile phenolic compound can be efficiently brought into contact with the target substance.

[0044] For example, the target substance and the phenolic compound are allowed to coexist in a suitable container.

The container is preferably a sealed container, but may not be a complete sealed container. Alternatively, the target substance and the phenolic compound are placed in the same room, for example, the same storage room, a display shelf, Coexist in the case etc.

[0045] Since the anti-oxidation agent that is widely used at present is used by adding to the target substance, it is necessary to consider the solubility in the target object. In addition, depending on the type and amount of the anti-oxidation agent, acidity and bitterness may be imparted and coloring may occur. Therefore, the method of the present invention in which the phenolic compound is allowed to coexist with the target substance in a volatilizable state can be a simpler method for preventing acidification than the conventional acid inhibitor that requires addition.

[0046] For example, a phenolic compound is usually present in an amount of 20 to 80 µmol, preferably 40 to 60 µmol, per lg of the subject substance under the condition that the phenolic compound strength is 25 ° C. In addition, the Ministry of Health, Labor and Welfare has indicated a provisional guideline value that the indoor volatile organic matter (VOC) concentration should be kept to 400 gZm ³ or less, so organic substances including phenolic compounds in volatiles It is desirable that the upper limit of the substance concentration is about 400 μgZm ³ .

[0047] The target substance for preventing acidification in the present invention is not particularly limited, and examples thereof include foods, pharmaceuticals, quasi drugs, cosmetics, and feeds. The present invention is particularly preferably used for the prevention of acidification of an oily component or a substance containing an oily component.

[0048] The oily component is not particularly limited and may be synthetic or natural. For example, hydrocarbons, straw steals, animal and vegetable oils and fats, waxes, goose fats, higher fatty acids, and higher alcohols , Silicone-based substances, sterols, greaves, etc., which have been treated enzymatically (hydrolysis, transesterification, etc.) or chemical treatment (transesterification, hydrogenation, etc.). Specifically, soybean oil, rapeseed oil, corn oil, sesame oil, cottonseed oil, safflower oil, sunflower oil, peanut oil, rice germ oil, wheat germ oil, brown rice germ oil, pearl barley oil, mackerel damian nut oil, garlic oil , Camellia oil, palm oil, olive oil, jojoba oil, mackerel demian nut oil, apogado oil, castor oil, flax oil, perilla oil, eucalyptus oil, evening primrose oil, turtle oil, mink oil, lard, beef tallow, horse Oil, snake oil, fish oil, egg oil, egg yolk oil, liquid paraffin, isono << raffin, petrolatum, squalene, squalene, turpentine oil, myristic acid isopropyl ester, myristic acid isopalmityl ester, myristic acid 2-octyldodecyl ester, 2 Ethylhexanoic acid cetyl ester, Trie 2-ethylhexanoic acid glyceryl ester, Tricaprylic acid glyceride A mixed fatty acid triglyceride, di-2-ethylhexanoic acid neopentyldarlicol ester , Malic acid diisostearyl ester, isononanoic acid isonol ester (3,5,5-trimethylhexyl 3,5 ', 5, trimethylhexanoate), 12 hydroxystearic acid cholesteryl ester, Emery Isostearic acid and Z or monoester of higher fatty acid and dipentaerythritol, hexaester, paramethoxy cinnamate and glycerin ester of 2-ethylhexanoic acid, isooctyl ester of para methoxy cinnamate, etc. Is mentioned.

[0049] Also, soybean hardened oil, rapeseed oil, palm hardened oil, fish hardened oil, Dali Tristearate Serinore Estenole, Rosin, Cholesterol Nore, Phytosterol Nore (Campesteronore, Stigmasterol, Sitosterol Etc.), orange luffy oil, lanolin, myristic acid, palmitic acid, isopalmitic acid, stearic acid, Emery's isostearic acid, oleic acid, linoleic acid, linolenic acid, ricinoleic acid, 12-hydroxystearic acid, 10-hydroxy Stearic acid, behenic acid, erucic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, myristyl alcohol, cetyl alcohol, stearyl alcohol, isostearyl alcohol, behanol alcohol, lanolin alcohol, paraffin wax, myChlocrystallin wax, ceresin wax, beeswax, petrolatum, hard fat, carnauba wax, candelilla wax, rice wax, rice bran wax, wax wax shellac, dimethylpolysiloxane, methylphenol polysiloxane, animal and plant essential oil components, etc. Can be targeted.

[0050] Examples of the substance containing an oily component include foods and drinks, pharmaceuticals, quasi drugs, cosmetics, and feeds containing an oily component. For example, ramen, tanmen, udon, buckwheat, macaroni, spaghetti, fried rice cake, non-fried rice cake, etc .; powder and liquid soup; milk such as yogurt, prepared milk, infant formula, ice cream, yogurt, cream, whipped cream Products; Rice crackers, snacks, biscuits, wafers, chocolates, strawberries, caramels, gums, candies, gummy and other sweets; Chikuwa, power mackerel, fish sausages, fish ham and other marine products; Frozen foods such as fried chicken, frozen fried food, frozen dumplings, frozen sweet potatoes; miso, hamburger, ham, sausage, dumplings, sweet potatoes, bread, cooking oil, butter, margarine, shortening, cheese, mayonnaise, dressing, beverage, health Food, treatment Other foods such as food; lipstick, cosmetic cream, milky lotion, shi Shampoo, rinse, knocking agent, Cosmetics such as cataplasms; pharmaceutical drinks and quasi-drugs such as energy drinks, powders, tablets, ointments and vitamins; industrial products such as fertilizers and lubricants; and feeds such as pet food .

The present invention also provides formula I:

[Chemical 5]

[0052] [where

R ¹ may have a substituent, may be C alkyl or have a substituent! /, May! /, C

1-6 1 Anoroxy

-6

1-6

Yo! /, C alkenyl, substituted! /, May! /, C alkynyl or substituted

2-6 2-6

/! Or even! /, C alkoxy,

1-6

R ^{3 to} R ⁵ are each independently a hydrogen atom or a substituent]

The present invention relates to an antioxidant containing a phenolic compound represented by the formula:

[0053] The compounds of formula I and preferred compounds are the same as described for the antioxidant method. The antioxidant of the present invention may contain one or more compounds as the compound of formula I. The antioxidant of the present invention is preferably for preventing the oxidation of oily components. The oil component is as described above.

[0054] In the anti-oxidation agent of the present invention, the phenolic compound is volatilized. In other words, the phenolic compound that has been vaporized into gas comes into contact with the target substance, and is in a state where it can exhibit the effect of preventing acidification.

[0055] The present invention also relates to a method for evaluating the antioxidant ability of a test substance in a volatile state.

[0056] In one embodiment, the present invention relates to a method for evaluating the antioxidant ability of a test substance in a volatile state. A) a step of measuring the absorbance of a solution containing linoleic acid and | 8-strengthen, b) a step of separately placing the solution and the test substance in a sealed container, c) a fixed time A step of measuring the absorbance of a solution containing linoleic acid and j8-strengthen after elapse, and d) a step of calculating a difference between the absorbance measured in step a and the absorbance measured in step c.

The method.

[0057] The above embodiment utilizes the fact that β-carotene reacts with the reaction of acidified linoleic acid and β-carotene over time. When the test substance is volatile and exhibits anti-oxidation ability, the volatilized test substance reacts with the β-carotene solution to suppress the fading of β-carotene. By judging the change in j8-carotene from the absorbance (absorbance at 490 nm), the antioxidant ability of the test substance can be evaluated.

[0058] It is preferable to further add a surfactant to the solution containing linoleic acid and β-strength. The surfactant is not particularly limited, but for example, Tween 40 can be used. As the solvent, water or ethanol can be used, and a buffer solution may be prepared if desired. Concentration of Reno one Honoré acid in the solution is usually 100 to 500 _8/1! 1, preferably 200~400 μ gZmL, concentration of β- carotene, usually 5 to 50 GZmL, preferably 10 to 40 mu gZmL. The pH of the solution is normally 5-8. The time for placing the solution and the test substance in the sealed container is appropriately selected, but is usually 0.5 to 3 hours, preferably 1 to 2.5 hours, and the temperature is usually 10 to 30 ° C. .

[0059] In the step b), the solution and the test substance may be introduced into another hole using, for example, a 96-well plate and placed in a sealed container. Disposing separately means that the solution and the test substance are not mixed or contacted.

[0060] In one embodiment, the present invention relates to a method for evaluating the antioxidative ability of a test substance to an oily component in a volatile state, comprising: a) an oily component to which linoleic acid and | 8-carotene are added. A step of measuring the absorbance, b) a step of separately placing the oily component and the test substance in a sealed container, c) a step of measuring the absorbance of the oily component after a certain period of time, and d) in step a. Calculating the difference between the measured absorbance and the absorbance measured in step c,

The method.

[0061] The above embodiment also reacts 13-carotene with linoleic acid that has been acidified over time, It uses the discoloration of carotene, and is different in that it uses an oily component with linoleic acid added. The oily component is not particularly limited, but may be appropriately selected from the above oily components and the like which are preferably liquids at the temperature to be tested, usually room temperature. For example, corn oil, soybean oil, rapeseed oil, sesame oil, cottonseed oil, safflower oil, sunflower oil, peanut oil, rice germ oil, wheat germ oil, brown rice germ oil, pearl barley oil, mackerel damian nut oil, garlic oil, camellia oil Vegetable oils such as palm oil and olive oil can be used.

[0062] In the above embodiment, the concentration of linoleic acid in the oily component is usually 0.1 to 6 mgZmL, preferably 0.5 to 5 mgZmL. gm is preferably 3 to 50 μg ZmL.

[0063] In one embodiment, the present invention relates to a method for evaluating the antioxidative ability of an analyte in an volatile state of a test substance, comprising: a) measuring the amount of malondialdehyde in the oil component; b) A step of separately placing the oily component and the test substance in a sealed container; c) a step of measuring the amount of malondialdehyde in the oily component after a certain period of time; and d) the amount of malondialdehyde measured in step a) Calculating the difference in the amount of malondialdehyde measured in steps c and c,

The method.

[0064] Malondialdehyde (MDA) is a secondary product of lipid peroxide and reacts with acid-conditioned thiobarbituric acid to form a red pigment. Lipid peracid can be measured by measuring the absorption wavelength of the resulting red pigment at 595 nm (Ohkawa H et al, Anal. Biochem., 95, 351-358 (1979), Kikukawa K et al "Anal Biochem., 202: 249-255 (19 92)).

[0065] In the above two embodiments using an oily component, the time for placing the oily component and the test substance in the sealed container is an appropriately selected force. Usually 4 to: L0 days, preferably 5 to 7 days. The temperature is usually 50-60 ° C. The evaluation method using the oil component is advantageous in that it can more accurately evaluate the antioxidant ability of the test substance against the oil component.

[0066] In the embodiment in which the anti-acid ability is evaluated by measuring the fading of the β-strength mouth by the difference in absorbance with the oily component added with linoleic acid and the β-strength mouth strength. And at the same time Marron You can measure changes in the amount of dialdehyde.

[0067] When the anti-acid ability is evaluated by measuring the fading of β-strength by the difference in absorbance, after 150 minutes at room temperature (25 ° C) in the presence of 500 mol of the test substance. If the difference in absorbance is 0.8 or less, preferably 0.7 or less, the test substance can be determined to have an antioxidant ability in a volatile state. Alternatively, if the difference in absorbance is 1.5 or less, preferably 1.2 or less after 5 days at 50 ° C in the presence of 500 mol of the test substance, the test substance is in a volatile state. It can be determined that it has antioxidant ability.

[0068] When the anti-acidic ability is evaluated by measuring the amount of malondialdehyde, control of the amount of malondialdehyde after 5 days at 50 ° C in the presence of 500 μmol of the test substance. If the ratio is 0.8 or less per lg of oily component, preferably 0.6 or less, the test substance can be determined to have antioxidant ability in a volatile state.

[0069] In the method for evaluating anti-acidic ability of the present invention, the test substance is not particularly limited and may be liquid or solid. For example, peptides, proteins, non-peptidic compounds, synthetic compounds, fermented products, cell extracts, plant extracts, animal tissue extracts, etc. are used, and these substances may be novel substances. It may be a known substance.

Example

[0070] Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to the scope of the examples.

[0071] Example 1

Equipment and testing

The 96-well plate used was TPP TISSUE CULTURE TEST PLATES 96F. As a sealed container, an anero pack jar (inside dimensions: 19.5 cm × 19.7 cm × l. 8 cm) manufactured by Sgiamagen Co., Ltd. was used. As a microplate reader, Nippon Bio-Rad (Model550) was used. All reagents were purchased from Wako Pure Chemical.

[0072] Preparation of sample solution

Using black mouth form, j8-carotene was prepared at lmg / mL, linoleic acid at 0.1 lg / mL, and Tween 40 at 0.2 gZmL. Using water, sodium dihydrogen phosphate was adjusted to a concentration of 24 g / mL (0.2 M), and disodium hydrogen phosphate was adjusted to a concentration of 28 g ZmL (0.2 M). Re Adjust the pH of sodium dihydrogen phosphate aqueous solution (0.2 M) to the disodium hydrogen phosphate aqueous solution.

6.8 and 0.2 M phosphate buffer.

[0073] | 8-Carotene solution (225 1 ^), linolenic acid solution (50 1 ^), and Ding 661140 (150 1 ^) were placed in a 50 mL Erlenmeyer flask, and the black mouth form was completely removed with nitrogen gas. After that, distilled water (15 mL) was dissolved. To this, 1.35 mL of 0.2 M phosphate buffer (pH 6.8) was added to prepare a linoleic acid 'β-carotene solution.

[0074] 200 L of linoleic acid 'β single-strength solution was added to the center of the 96-well plate. Phenolic compounds (buluguaiacol, isoeugenol, eugenol, guaiacol, 2, 3, 6-trimethylphenol) and other test substances (each 500 mol) were added to the four corners of a 96-well plate ( Figure 1).

[0075] Immediately thereafter, the absorbance (A) at 490 nm was measured with a microplate reader.

0

. After A measurement, place the 96-well plate in a sealed container (Anero pack jar) and seal it (Fig. 1).

0

). After t minutes, the amount of decrease in absorbance at 490 nm, that is, Δ A = A−A was calculated. T o t

The same operation was carried out without using any phenolic compound or test substance.

[0076] Figure 2 shows the fading of j8-carotene (decrease in absorbance = 0, 30, 60, 90, 120, and 150 minutes after addition of burguaiacol (PVG) or tocopherol (Toe). ΔΑ

)showed that. It can be said that the smaller the ΔΑ, the stronger the acid resistance.

[0077] It was shown that the phenolic compound of the present invention has an excellent antioxidant power in a volatile state.

[0078] Example 2

Equipment and testing

The 96-well plate used was TPP TISSUE CULTURE TEST PLATES 96F. As a sealed container, an anero pack jar (inside dimensions: 19.5 cm × 19.7 cm × l. 8 cm) manufactured by Sgiamagen Co., Ltd. was used. As a microplate reader, Nippon Bio-Rad (Model550) was used. Reagents were purchased from Wako Pure Chemical.

[0079] Preparation of sample solution

Using Kuroguchi Form, the concentration of β-strength mouth is lmgZmL and linoleic acid is 0.lgZmL [0080] The β-carotene solution (240 L) and linoleic acid solution (480 L) were placed in a test tube with a lid, and the black mouth form was completely removed with nitrogen gas. Then, tocopherol-removed corn oil (6m

L) The mixture was stirred and a linoleic acid 'β-carotene solution was prepared.

[0081] 100 L of linoleic acid 'β-carotene solution was added to the center of a 96-well plate. Phenolic compounds (buluguaiacol, isoeugenol, eugenol, guaiacol) were added to the four corners of the 96-well plate (Fig. 1).

[0082] Immediately thereafter, the absorbance (Α) at 490 nm was measured with a microplate reader.

0

) o

[0083] After t days, the amount of decrease in absorbance at 490 nm, that is, Δ A = A−A was calculated. T o t

The same operation was carried out for the trawl without using any phenolic compound or test substance. It can be said that the smaller the ΔΑ, the stronger the acidity.

[0084] After 5 days, the amount of malondialdehyde in the linoleic acid 'β-strength tenten solution was measured. The acetic acid buffer was prepared by dissolving 20 mL of acetic acid in water to make 10 mL, and adjusted to pH 3.5 with ION NaOH. The 8% TBA solution was heated and then subjected to ultrasound to completely dissolve the reagent.

[0085] In a test tube with a screw cap, 0.2 g of linoleic acid 'β-strength tenten solution, 8.1 mL of an aqueous solution of sodium dodecyl sulfate (SDS) 0.2 mL, acetate buffer 1.5 mL, 0.8% Butyl hydroxytoluene (BHT) acetic acid solution 0.05 mL, 0.8% TBA solution 1.5 mL, and water ImL were mixed well and mixed well. Sealed and left at 5 ° C for 60 minutes, and then heated in a 100 ° C block heater for 60 minutes. After cooling, 1 mL of water and 5 mL of butanol / pyridine (15Z1) were added and mixed well, followed by centrifugation (3000 rpm, 10 minutes). The resulting supernatant was measured for absorbance at 532 nm. The molecular extinction coefficient (ε = 156000) was used to convert to malondialdehyde (MDA) and divided by the control for comparison. Separately, a sample blank was prepared by performing the same operation for those to which no oil was added.

[0086] FIG. 3 shows the amount of decrease in absorbance after 5 days when each substance is added, that is, the amount of fading of β-strength. When the phenolic compounds bull guaiacol, isoeugenol, eugenol, and guaiacol were volatilized in a sealed container, they showed strong anti-acid activity. Other substances used for comparison have no effect, and this antioxidant It was proved that the effect was unique to phenolic compounds.

[0087] Fig. 4 shows the ratio of malondialdehyde to control after 5 days when each substance was added. When the phenolic compounds bull guaiacol, isoeugenol, eugenol, and guaiacol were volatilized in a sealed container, they showed strong antioxidant capacity. The other substances used for comparison did not show any action, and it was found that this acid / anti-oxidation effect was peculiar to phenolic compounds.

[0088] All publications, patents, and patent applications cited herein are incorporated herein by reference in their entirety.

Claims

The scope of the claims

[1] An antioxidant method that involves allowing a phenolic compound to coexist with a target substance in a volatilizable state.

[2] Phenolic compounds have the following formula I:

[Chemical 1]

[Where

R ¹ may have a substituent, may be C alkyl or have a substituent! /, May! /, C

1-6 1 Anoroxy

-6

1-6

Yo! /, C alkenyl, substituted! /, May! /, C alkynyl or substituted

2-6 2-6

/! Or even! /, C alkoxy,

1-6

R ^{3 to} R ⁵ are each independently a hydrogen atom or a substituent]

The antioxidant method according to claim 1, which is a compound represented by the formula:

[3] The antioxidant method according to claim 1 or 2, wherein the target substance is an oily component or a substance containing an oily component.

[4] The method according to any one of claims 1 to 3, wherein the phenolic compound is at least one selected from the group force of guaiacol, bulguaiacol, eugenol, isoeugenol and 2,4,6-trimethylphenol. How to prevent oxidation.

[5] The following formula I:

[Chemical 2]

[Where

R ¹ may have a substituent, may be C alkyl or have a substituent! /, May! /, C

1-6]

-6

Yo! /, C alkenyl, substituted! /, May! /, C alkynyl or substituted

2-6 2-6

/! Or even! /, C alkoxy,

1-6

R ^{3 to} R ⁵ are each independently a hydrogen atom or a substituent]

The antioxidant containing the phenolic compound represented by these.

[6] The antioxidant according to claim 5, for preventing acidity of the oily component or the substance containing the oily component.

[7] The antioxidant according to claim 5 or 6, comprising at least one selected from the group power of guaiacol, bulguaiacol, eugenol, isoeugenol and 2,4,6 trimethylphenol.

[8] The antioxidant according to any one of claims 5 to 7, wherein the phenolic compound is in a volatizable state.

[9] A method for evaluating the antioxidant capacity of a test substance in a volatile state,

[10] A method for evaluating an antioxidant ability of a test substance against an oily component in a volatile state, a) measuring the absorbance of the oily component to which linoleic acid and 13-carotene are added; b) placing the oily component and the test substance separately in a sealed container;

A method for evaluating the antioxidant ability of a test substance against an oily component in a volatile state, a) a step of measuring the amount of malondialdehyde in the oily component;

Including said method.