JP2000072686A - Active-oxygen scavenger and composition including the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To prepare an active-oxygen scavenger having high active-oxygen- scavenging potency and high antioxidative potency, and sparingly having a side-effect on the human body and having safety for the human body by including an extract of seeds (or their husks) of a plant belonging to the geneses Pecan in family Juglandaceae. SOLUTION: This active-oxygen scavenger includes an extract of seeds (or their husks) of a plant belonging to the genus Pecan in family Juglandaceae. The extract is pref. prepared by extraction with a solvent or a mixed solvent selected from the group consisting of water, methanol, ethanol, n-propanol, iso-propanol and acetone. It is pref. that the quantity of the extract added to food is 0.03-2 wt.% based on the total weight when the food is a seasoning such as fermented soybean paste, soy sauce, mayonnaise, or the like and cooking oil such as salad oil, and is 0.02-2 wt.% based on the total weight when the food is other than those above mentioned. The quantity of the extract added to feed is pref. 0.02-2 wt.% based on the total weight.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an active oxygen scavenger and a composition containing the same, and more particularly to an active oxygen scavenger containing an extract of seeds or seed husks of a plant belonging to the genus Pecan of the family Walnut, and an active oxygen scavenger containing the same. The present invention relates to a composition containing an active oxygen scavenger.

[0002]

2. Description of the Related Art Oxygen is an essential substance for the living body, such as the production of energy by metabolism, for maintaining life. However, oxygen is reactive due to various reaction systems inside and outside the living body, that is, some immune reactions, ultraviolet rays and radiation irradiation. rich reactive oxygen species (superoxide anion: O ₂ ^-, hydrogen peroxide: H ₂ O _2, the hydroxyl radical: OH ·, singlet oxygen: ¹ O _2, etc.) to produce. While these reactive oxygen species are useful for inactivating foreign substances such as invading bacteria in certain immune reactions, for example, they react with biological components to cause lipid peroxidation and denaturation of proteins and nucleic acids, It is known that it causes various diseases and aging. In particular, the skin is in the outermost layer of the body, so it is susceptible to reactive oxygen species generated by external factors such as ultraviolet rays and radiation. It is known that the production of abnormal skin pigments such as spots and freckles is enhanced.

[0003] O ₂ caused by various causes in vivo ^- is
It is converted to H ₂ O ₂ by superoxide dismutase (hereinafter abbreviated as SOD), and H ₂ O ₂ is further decomposed into water and oxygen by the action of catalase or glutathione peroxidase. In addition to these enzymes, there are food-derived antioxidants such as vitamin C and vitamin E in the living body, which form a network that suppresses radical and oxidative reactions in the living body and cause various obstacles due to active oxygen. It is believed to be the defense of However, changes in the surrounding and in vivo environment, such as infection by bacteria and viruses,
Changes in the state of food intake and nutrition, excessive irradiation of ultraviolet rays, various stresses from the surroundings, and aging and aging, etc., disrupt the above-mentioned network balance, and the metabolic balance of the active oxygen produced in the living body may be lost. Collapse, resulting in increased lipid peroxide levels, dermatitis, spots, wrinkles, eczema,
It is known that various cosmetically disturbing symptoms such as freckles appear, and furthermore, diseases such as rheumatoid arthritis, arteriosclerosis, diabetes, hepatitis, nephritis and cancer are caused.

[0004] In addition, the foods and food additives we ingest are also oxidized by active oxygen generated by the action of ultraviolet rays or radiation or by normal oxidation, resulting in the peroxidation or denaturation of lipids and other components contained therein. It is well known that ingestion of these oxidized and denatured products is not preferable for health. Furthermore, it is known that some of the natural fats and oils and surfactants containing unsaturated lipids which may be added to cosmetics and external preparations for skin are susceptible to oxidation by the action of ultraviolet rays and the like. Undesirable phenomena such as discoloration and generation of an unusual odor often occur. The use of such a cosmetic base which is susceptible to oxidation easily increases the amount of lipid peroxide generated in the skin due to the above-mentioned causes, and it is easy to amplify the above-mentioned various symptoms which are cosmetically impaired. It is to be guessed.

[0005] Natural oils and fats which are susceptible to oxidation include:
Unsaturated oils and fats such as almond oil, olive oil, jojoba oil and squalene, and surfactants that are easily oxidized include unsaturated surfactants such as glyceryl monooleate and polyoxyethylene sorbitan trioleate. No. Conventionally, antioxidants added to cosmetics, foods, food additives and feeds include synthetic antioxidants such as dibutylhydroxytoluene (BHT), butylhydroxyanisole (BHA) and ethoxyquin, ascorbic acid and vitamins. There are natural antioxidants such as E. Although the above-mentioned synthetic antioxidants have excellent antioxidant effects, there are some which have a problem in safety such as carcinogenicity and the use thereof is restricted in some cases. On the other hand, the above-mentioned natural antioxidants are evaluated for safety, but have a drawback that their antioxidant effects are considerably inferior to synthetic antioxidants.

[0006] In recent years, in order to improve and prevent the above phenomena,
Active search for cosmetics, food science, and pharmacological aspects of substances or compositions with active oxygen scavenging or antioxidant effects has resulted in a significant number of active oxygen scavengers and antioxidants. Agents became known. For example, in Japanese Patent Publication No. 4-34969, an active oxygen scavenger containing baicalein, which is a flavonoid component contained in Japanese gougon, is disclosed in Japanese Patent Application Laid-Open No. 3-227938, from clove oil or dehydrodioigenol, which is a component thereof. In Japanese Patent Application Laid-Open No. 5-271630, an active oxygen scavenger / removal agent containing an asparasas linearis extract as an active ingredient is disclosed in JP-A-7-6991.
No. 2 proposes an active oxygen scavenger containing a walnut shell extract as an active ingredient.

Further, cosmetics and foods containing a component having an active oxygen scavenging effect have been proposed.
JP-A No. 2556 discloses an external preparation for skin obtained by further adding SOD to a ginkgo biloba extract having an active oxygen removing effect.
In Japanese Patent Application Laid-Open No. 6-65043, cosmetics and foods containing an extract component of Shirakashi as an active ingredient are one or more of plant extracts selected from Maikaika, Mokka and Ichinin having an active oxygen removing effect. Japanese Patent Application Laid-Open No. 6-183987 discloses a skin external preparation characterized by containing a SOD-containing peroxidative extract comprising a plant extract of the genus Pelargonium having a high active oxygen removing effect and a lipid peroxide production inhibitory effect. A lipid production inhibitor and a composition (cosmetics, foods, pharmaceuticals) containing the same as an active ingredient are proposed, and JP-A-7-213251 proposes health foods and drinks to which an antioxidant extracted from cacao beans is added. ing.

[0008]

SUMMARY OF THE INVENTION An object of the present invention is to provide a highly safe active oxygen scavenger which has extremely high active oxygen scavenging ability and antioxidant ability, has few side effects on the human body, and has high safety. Is to provide a composition (food, feed or cosmetics) containing The present inventors have conducted intensive studies to solve the above-mentioned problems, and as a result, the seeds or seed husks of plants of the genus Pecan of the family Walnuts, which are used as edible water, are heated with hot water, methanol, ethanol, propanol, and acetone. The present invention has been completed by finding that a solvent and an extract obtained by extraction with a mixed solvent thereof have very strong active oxygen scavenging ability and antioxidant ability, and that the extract has high safety.

[0009]

That is, the present invention according to claim 1 provides a plant belonging to the genus Juglandaceae Pecan (Cary).
It is an active oxygen scavenger containing a seed or a seed shell extract of a plant belonging to a). According to a second aspect of the present invention, the extract according to the first aspect comprises water, methanol, ethanol, n-
The active oxygen scavenger according to claim 1, wherein the active oxygen scavenger is extracted with one or more mixed solvents selected from propanol, iso-propanol and acetone. According to a third aspect of the present invention, there is provided a food, feed or cosmetic containing the active oxygen scavenger according to the first aspect.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail. The plant of the genus Pecan of the walnut family used in the present invention,
It is mainly distributed from North America to Central America, and the embryo part of its seeds has been used for food since ancient times, and the seeds themselves are called pecan nuts or pecan nuts. The active oxygen scavenger of the present invention may be used alone or in the form of an organic solvent such as water, hot water, methanol, ethanol, n-propanol, iso-propanol, acetone, etc. And a mixed solvent of an organic solvent and a mixed solvent, preferably an extract obtained by extraction with a hydrate of the above organic solvent as an active ingredient.

[0011] The extraction method is not particularly limited.
What is necessary is just to carry out according to a usual method. Hereinafter, a preferred extraction method will be described. The amount of the extraction solvent used is not particularly limited, and may be usually 5 to 100 times the weight of the pecan nut or its shell used. The extraction temperature may be about 20 ° C. to 120 ° C. when water or hot water is used as the extraction solvent. When an organic solvent or a water-containing organic solvent is used as the extraction solvent, it is not particularly limited, but preferably at room temperature, particularly preferably 20
Extract at a temperature of between 30C and 30C. The extraction time may be about 5 to 60 minutes in the case of water or hot water extraction, but 24 to 96 hours is appropriate in the case of extraction with an organic solvent or a water-containing organic solvent. After the extraction, the extraction solvent-soluble portion is obtained by means of natural filtration using filter paper or absorbent cotton, filtration under reduced pressure, and the like, and then the solvent is distilled off by means of concentration under reduced pressure, and further frozen if necessary. The active ingredient of the present invention is obtained by drying or spray drying.

The active ingredient of the present invention thus obtained has an extremely high active oxygen scavenging effect and an antioxidant effect, is highly safe, and its effects are not lost even if processing means such as heating are added. Since it is stable, a stable and useful active oxygen scavenger or antioxidant can be obtained by incorporating the active ingredient of the present invention. Further, the active oxygen scavenger or antioxidant of the present invention may be prepared by blending the active ingredient itself in a composition such as food or cosmetics, or blending the active ingredient in advance into the composition. In addition, it is possible to enhance the commercial value of the composition by imparting an active oxygen scavenging effect and an antioxidant effect.

In the case of tablets, capsules, powders or granules, examples of the formulation include the above active ingredients as excipients such as starch, lactose and mannitol, binders such as carboxymethylcellulose and hydroxypropylcellulose, and crystalline cellulose. And disintegrants such as calcium and carboxymethylcellulose, lubricating agents such as talc and magnesium stearate, and, if necessary, wetting agents, coloring agents, flavors, and the like. In addition, the liquid preparation may be an aqueous or oily emulsion or syrup, a simple syrup, sorbitol, a suspending agent such as methylcellulose or sodium carboxymethylcellulose, egg yolk lecithin, sorbitan monofatty acid ester, lauromacrogol, An emulsifier such as castor oil, and if necessary, a preservative, a preservative, a stabilizer and the like can be appropriately compounded to form a preparation. In the case of ointments, petroleum bases such as petrolatum, paraffin, silicone, plastibase, vegetable oils and waxes, purified lanolin, carboxyvinyl polymers, hydrophilic bases such as propylene glycol and 1,3-butanediol, polyethanolamine And the like can be formulated by appropriately mixing an emulsifier and the like.

Among the compositions of the present invention, various forms of foods can be mentioned. For example, they can be added to seasonings such as miso, soy sauce, mayonnaise, etc., and edible oils such as salad oil.
In addition to mixing, various cooked foods; desserts, frozen desserts, candy, sweets and drinks such as chewing gum and fruit juice, and the like can be mentioned. In these foods, any components according to the purpose of use can be used. For example, in the case of frozen desserts and beverages, fruit juices, sweeteners, sour agents, coloring agents, flavors, and the like can be appropriately compounded. The amount of the active ingredient of the present invention to be added to foods may be appropriately changed depending on the form. For example, miso, soy sauce, seasoning such as mayonnaise and edible oil such as salad oil have a total amount of 0.03 to 2% by weight. %, And 0.02 to 2% by weight for other foods.

[0015] Among the compositions of the present invention, the form of the feed can be various, and for example, a powdered, pasty or pelletized compound feed for livestock, poultry and fish can be mentioned. These feeds can use arbitrary components according to the purpose of use. For example, in the case of a feed in the form of a kneaded product, a colorant, a flavor, and the like can be appropriately compounded. The amount of the active ingredient of the present invention added to the feed may be appropriately changed depending on the form, but generally, the total amount is 0.02%.
It is preferably set to と す る 2% by weight.

Among the compositions of the present invention, various forms of cosmetics can be mentioned, for example, lotions,
Examples include facial cleansers, packs, makeup cosmetics, hair cosmetics, lip cosmetics, nail cosmetics, bath salts, antiperspirants, and the like, including emulsions and creams. These cosmetics can use any components according to the purpose of use,
For example, in the case of cream, hydrophobic bases such as petrolatum, paraffin, and squalane; hydrophilic bases such as lanolin, propylene glycol and 1,3-butanediol; emulsifiers such as fatty acid monoglycerides and sorbitan fatty acid esters; , A pigment, a fragrance, and, if necessary, a nutrient, a humectant, a whitening agent, an ultraviolet absorber and the like can be appropriately compounded according to a conventional method. Similarly, for other products, necessary components according to the types thereof can be appropriately compounded. The amount of the active ingredient of the present invention added to cosmetics is
It may be changed appropriately depending on the form, but it is preferably about 0.02 to 2% by weight of the whole cosmetic.

As described above, the active oxygen scavenger of the present invention can be used by adding it to compositions such as foods, feeds, and cosmetics. To improve the stability of these compositions, and also to consumers or users of these compositions,
It is very useful because it can protect against various disorders caused by active oxygen and lipid peroxide.

[0018]

EXAMPLES Next, the present invention will be described in more detail with reference to examples and the like, but the present invention is not limited thereto. Production Example 1 Seed shells obtained by removing the edible portion of pecan nuts were crushed,
500 ml of 50% aqueous ethanol is added to 100 g of the pulverized material.
(Milliliter, the same applies hereinafter), and the mixture was allowed to stand at room temperature for 48 hours for extraction. The obtained aqueous ethanol-soluble portion was filtered through a filter paper, concentrated under reduced pressure at 40 ° C. to distill off ethanol and water, and further freeze-dried to obtain the desired active ingredient (7.2 g of brown powder, yield: 7.2%).

Production Example 2 Pecan nuts were pulverized as they were, and 100 g of the pulverized products were added with 500 ml of 50% by volume aqueous ethanol, and extracted at room temperature for 48 hours. The obtained aqueous ethanol-soluble portion was filtered through filter paper, concentrated under reduced pressure at 40 ° C. to distill off ethanol and water, and further freeze-dried to obtain the desired active ingredient (7.4 g of brown powder, yield: 7.4%).

Production Example 3 500 ml of distilled water was added to 100 g of the pulverized seed hull obtained in the same manner as in Production Example 1, and the mixture was heated and extracted at 110 ° C. for 10 minutes. After cooling to room temperature, the water-soluble part was
The solution was concentrated under reduced pressure at 0 ° C. to distill off water, and further freeze-dried to obtain the desired active ingredient (6.5 g of light brown powder, yield 6.5).
%).

The infrared absorption spectrum of the active ingredient obtained by each of the above Production Examples was as follows. In addition, this spectrum measuring apparatus is IR-810 manufactured by JASCO Corporation.
The mold and sample preparation were performed by the KBr method. Absorption peak of active ingredient obtained in Production Example 1 3400-3300 (broad), 2925, 161
5, 1450, 1040, 830 cm ^-1 Absorption peak of active ingredient obtained by Production Example 2 3400 to 3300 (broad), 1615, 155
0, 1450, 1350, 1200, 840 cm ^-1 The absorption peak of the active ingredient obtained in Production Example 3 3400-3300 (broad), 1615, 145
0, 1350, 1200, 1040, 830 cm ^-1

[0022] Test Example 1 superoxide (O ₂ ^-) O ₂ of the active ingredient of the erase test the present invention ^- C.Beauchamp's method erase effect (Analytycal Biochemistry, vol.44, pp.276-287 ( 19
Nitro blue tetrazolium (NB)
T) It was examined by a reduction method. This method uses hypoxanthine (HP
This method utilizes the fact that O ₂ ⁻ generated when xanthin oxidase (XOD) is allowed to act on X) reduces NBT to produce dark blue formazan. O ₂ in this system ^-
In the presence of a substance having an erasing effect (sample = active ingredient of the present invention), the amount of formazan produced decreases, and the formazan production inhibition is calculated from the amount of formazan produced when the active ingredient is added to the amount of formazan produced when no active ingredient is added. obtains the rate, O ₂ of the active ingredient of the present invention ^- was erasure rate. Further, in this method, when the sample has the effect of inhibiting the enzymatic action of XOD is, O ₂ ^- even if no erasing effect, O ₂ ^-
Since the erasure rate becomes high apparent, rather than inhibitory effects of the active ingredient of the present invention is XOD, O ₂ ^- to confirm that it has an erase effect, the XOD inhibition of the active ingredients also investigated.

[0023] <O ₂ ^- method of measuring the erasing ratio> 50 mM (millimolar, hereinafter the same) sodium carbonate buffer (pH 1
0.2) 3mM HPX aqueous solution 0.1m in 2.4ml
1, 0.75 mM NBT aqueous solution 0.1 ml, 3 mM
0.1ml of EDTA (ethylenediaminetetraacetic acid) aqueous solution
And 0.1 ml of 0.15% bovine serum albumin aqueous solution
A solution of a sample (a 50% aqueous ethanol solution containing 0.3% by weight of the active ingredient of the present invention obtained in each Production Example) was added to a solution consisting of: The mixed solution thus added was placed in a cuvette (absorption wavelength measuring cell), and 0.1 ml of an XOD solution diluted to 0.05 U (enzyme unit) with purified water was added to start the reaction. The absorbance (A) at 560 nm was measured with a spectrophotometer (UVDEC430B, manufactured by JASCO Corporation) to determine the amount of formazan produced. As a control, the absorbance (B) was measured in the same manner using a mixed solution to which the same amount of 50% aqueous ethanol was added instead of the sample solution. Further, as a blank of the sample mixture, the sample mixture, likewise absorbance also reaction solution was added XOD solution was heated to inactivate the (C) was measured, O ₂ of the active ingredient of the present invention by the following equation 1 ^- was calculated erasure rate.

[0024]

(Equation 1)

[0025] <XOD inhibitory rate measurement method> The O ₂ ^- with a mixture of purified water was added to 0.1ml in place of 0.75 mM NBT solution during erase ratio measurement method, similar to the above method the XOD solution Three minutes after the addition, the absorbance (X) at 290 nm was measured to determine the amount of uric acid produced when HPX was oxidized by XOD. As a control, the absorbance (Y) at 290 nm was measured in the same manner using a mixed solution to which the same amount of 50% aqueous ethyl alcohol was added instead of the sample solution. Furthermore, the absorbance (Z) of the reaction mixture obtained by adding a heat-inactivated XOD solution to the specimen mixture as a blank of the specimen mixture was measured in the same manner.
The XOD inhibition rate of the active ingredient of the present invention was calculated. The results are shown in Table 1.

[0026]

(Equation 2)

[0027]

[Table 1]

As can be seen from Table 1, the active ingredient of the present invention is not limited to XO
Without substantially inhibiting the enzymatic action and D, a very high O ₂ ^-
Obviously, it shows an erasing effect.

Test Example 2 Hydroxyl radical (OH
-) Elimination test The OH-elimination effect of the active ingredient of the present invention was determined by the deoxyribose method (Analytical Biochemistry, vo
l.165, pp.215-219 (1987)) by a partially modified method. This method uses Fenton's reagent (hydrogen peroxide and iron [I
I] OH. Generated from an oxidative color reagent mixed with a salt)
Reacting thiobarbituric acid (TBA) with malondialdehyde (MDA) produced by the reaction of aldehydes and deoxyribose to form a red reactant, and measuring the amount of the produced red light at 532 nm to measure the absorbance (TBA value). It is what is sought. The presence of a substance having an OH / elimination effect (specimen = active ingredient of the present invention) in this system reduces the amount of MDA produced, and thus the TBA value. Therefore, the TBA value of the present invention when no active ingredient is added The MDA production inhibition rate was determined from the TBA value at the time of addition of the active ingredient of the present invention to the OH / elimination rate of the active ingredient of the present invention.

<Method of measuring OH / erasing rate> 200 mM
Potassium dihydrogen phosphate-potassium hydroxide buffer (pH
7.4) 0.2 ml, 28 mM deoxyribose aqueous solution 0.2 ml, 1 mM ferric chloride aqueous solution 0.2 m
1, 1 mM aqueous solution of ascorbic acid 0.2 ml,
0.2 ml of 04 mM EDTA aqueous solution, purified water
6 ml, (the solution was diluted with water such that the final concentration of mannitol was used as an active ingredient and positive controls of the present invention obtained in Production Example 1 is 200ppm) 10mM H ₂ 0 ₂ 0.2ml and sample solution 0.2ml Was reacted at 37 ° C. for 1 hour, 2 ml of a 20% aqueous solution of trichloroacetic acid and 1 ml of a 0.67% aqueous solution of TBA were added, and the mixture was heated in a boiling water bath for 10 minutes. After allowing the solution to cool to room temperature, the absorbance (E) at 532 nm was measured with a spectrophotometer (UVDEC 430B, manufactured by JASCO Corporation) to determine the TBA value. As a blank of the sample mixture, absorbance (F) was similarly obtained using a mixed solution to which purified water was added instead of 10 mM H ₂ O ₂ , and absorbance (G) when purified water was added instead of the sample solution as a control. Further, as a control blank, the absorbance (H) when purified water was added instead of the sample solution and purified water was added instead of 10 mM H ₂ O ₂ was measured. The OH elimination rate of the positive control was calculated. Table 2 shows the above results.

[0031]

(Equation 3)

[0032]

[Table 2]

As can be seen from Table 2, it is clear that the active ingredients of the present invention have a very high OH-erasing effect.

Test Example 3 Lipid Peroxidation Inhibition Test The lipid peroxidation inhibitory effect of the active ingredient of the present invention was evaluated by A. Jitoe
(Journal of Agricultural and Food Chemistr
y, vol. 40, pp. 1337-1340 (1992)), and examined by a thiocyanic acid method using linoleic acid as a lipid. This method utilizes the fact that ferrous ions are oxidized to ferric ions by peroxide of linoleic acid, and is a method of colorimetrically forming the generated ferric ions as thiocyanate complex salts.

<Method of Measuring Lipid Peroxidation Inhibition Rate> 99.5
% Ethanol 4 ml, 2.53 wt% linoleic acid solution in 99.5% ethanol 4.104 ml, 50 m
The reaction mixture obtained by adding 4 mg of each of the samples obtained in Production Examples 1 and 2 to 8 ml of M phosphate buffer (pH 7.0) and 3.896 ml of distilled water was placed in a 50-ml vial with a screw cap. After applying a screw cap, it was left in a thermostat set at 40 ° C. in the dark. As a positive control, a reaction mixture in which 4 mg each of dibutylhydroxytoluene and α-tocopherol were added instead of the active ingredient of the present invention was prepared. Eight days after the start of the experiment, 75%
9.7 ml of aqueous ethanol and 0.1 ml of a 30% aqueous solution of ammonium thiocyanate were added. 3.5% for this
0.1m of 0.02M ferrous chloride solution dissolved in hydrochloric acid water
1 and the absorbance (I) at 500 nm was measured exactly 3 minutes later. Further, as a blank, the absorbance (J) when the active ingredient of the present invention or the positive control was not added was measured, and the lipid peroxidation inhibition rate of each ingredient was calculated from the following equation 4. The results of the above experiments are shown in Table 3.

[0036]

(Equation 4)

[0037]

[Table 3]

As can be seen from Table 3, it is clear that the active ingredient of the present invention has a very strong lipid peroxidation inhibitory action.

Comparative Example The active ingredient of the present invention is an extract of seeds or seed husks of a plant belonging to the genus Pecan of the family Walnuts. On the other hand, as described above, it has been revealed that walnut shell extracts such as walnuts such as walnuts and walnuts exhibit an active oxygen elimination effect (JP-A-7-69912). Therefore, walnut; for (Walnut walnut) extract and shell extract of pecan nut of the present invention, their yield, O ₂ ^- was examined for the presence of differences in the erasure rate and components.

(1) Yield of Extract The shell obtained by removing the edible part of the pecan nut and the shell obtained by removing the edible part of the walnut were separately pulverized to obtain each pulverized material. Divide the crushed pecan nut shell into two,
100 g of each was placed in a 1-L Erlenmeyer flask, and one of them was added with 50% aqueous ethanol and the other with 500 ml of distilled water. The ground walnut shell was similarly divided into two parts, and 100 g of each was placed in a 1-L Erlenmeyer flask, and one of them was added with 50% aqueous ethanol and the other with 500 ml of distilled water. Each pulverized product to which 50% aqueous ethanol was added was allowed to stand at room temperature for 48 hours, then filtered to remove the residue, and the resulting extract was concentrated under reduced pressure at 40 ° C., and then freeze-dried overnight to obtain pecan nut shell. And a 50% aqueous ethanol extract of walnut shells.
For each pulverized product to which distilled water was added, 30 ° C. at 110 ° C.
After extraction with heating for 1 minute, the extract obtained by filtering to remove the residue was concentrated under reduced pressure at 40 ° C., and then freeze-dried overnight to obtain a hot water extract of pecan nut shell and a walnut shell hot water extract. Table 4 shows the yield of each extract relative to each crushed shell.

[0041]

[Table 4]

From Table 4, it can be seen that in the extraction with both solvents, the yield of pecan nut shell extract is 2.7 to 3 times higher than that of walnut shell extract. O ₂ of each extract was obtained by elimination activity above ^{(1) - - (2)} O 2 of each extract was assayed by the erase active nitroblue tetrazolium reduction method, without the addition of ED ₅₀ values (the sample of each extract when the production rate was 100%, O ₂ ^{- -} from the absorbance O ₂ obtained when seeking concentration of analyte) required to make the 50% scavenging activity compared active strength of the extract did. The ED ₅₀ value of each extract is as shown in Table 5.

[0043]

[Table 5]

[0044] As shown in Table 5, samples O ₂ ^- by determining from ED ₅₀ values indicating the strength of the scavenging activity, ED ₅₀ values than extract shell it is walnut extract shells pecan nut is 3.2 to 4.4 times lower, strong O ₂ a much ^- it is clear that having an erase activity.

(3) Components of each extract The hot water extract of the pecan nut shell obtained in Production Example 3 and the hot water extract of the walnut shell produced according to the same example are as follows:
Whether or not there was a difference between these components was examined by thin-layer chromatography (TLC method) and activated carbon adsorption method. TLC method The TLC measurement conditions are as shown below. Thin layer plate: Silica gel 60 F254
(MERCK) Developing solvent: 1-butanol: acetic acid: water = 6: 3: 1 Developing: Primary Developing distance: 12 cm Sample concentration: 300 ppm Application amount: 5 μl / spot Detection: UV (254 nm and 302 nm) and iodine coloring

The results of the TLC method for the hot water extract of pecan nut shell and walnut shell are shown in Table 6.

[0047]

[Table 6]

As shown in the table, the hot water extract of the pecan nut shell was divided into three components, while the hot water extract of the walnut shell was divided into two components. Based on the difference in each Rf value, it is determined that the components of these extracts are different from each other.

Activated carbon adsorption method The hot water extract of pecan nut shell and the hot water extract of walnut shell are dissolved in water so as to have the same concentration (0.3% [weight / volume]). Twice (weight) activated carbon (activated carbon (powder)) (untreated: manufactured by Nacalai Tesque) was added, and the mixture was stirred at room temperature for 4 hours. Thereafter, each treatment liquid was filtered using a filter paper (5C type: manufactured by Toyo Roshi Kaisha, Ltd.), and the filtrate was concentrated under reduced pressure and freeze-dried. The resulting freeze-dried product of the filtrate to be 0.3%, respectively (weight / volume) dissolved in water, O ₂ of each solution ^- was measured scavenging activity. As a control, an aqueous solution (0.3% [weight / volume]) before the activated carbon treatment was used. O ₂ of each solution ^- measurement of the scavenging activity is as shown in Table 7.

[0050]

[Table 7]

[0051] As is clear from Table, O ₂ lyophilisate of the filtrate after the activated carbon treatment of hot water extract of pecan nut shell ^- erasure rate is changed scarcely observed as compared to control, walnut shells O ₂ of the hot water extract of the freeze-dried product of the filtrate after the activated carbon treatment
^- erasing rate, it has decreased compared to the control. Therefore,
Pecan nut O hot water extract of the shell ₂ ^- scavenging activity component, whereas not the component to lose active by treatment with activated carbon, O ₂ of hot water extract of walnut shell ^- Clear active ingredient, said active by treatment with activated carbon Of these extracts from losing
O ₂ ^- scavenging activity ingredient is determined to be different components.

Test Example 4 As a safety test of the active ingredient of the present invention, acute toxicity test (rat) and mutagenicity test (Salmonella typhimurium) TA 98, TA 100, TA 1
535, TA 1537 and Escherchia coli WP2
Ames test using uvra), skin sensitization test (guinea pig and human patch test), primary skin irritation test (guinea pig) and phototoxicity test (guinea pig) were performed.

(1) Acute toxicity test by single oral administration Production Example 2 was conducted on 5 male Wistar rats weighing 180 g.
When the active ingredient of the present invention obtained in the above was orally administered so as to be 2 g / Kg (body weight), and then normally bred for 2 weeks,
Mortality was 0%. Therefore, the half-lethal dose (LD ₅₀ ) of the active ingredient of the present invention was 2 g / Kg or more, and the toxicity was determined to be extremely low.

(2) Mutagenicity test An Ames test was performed using each of the above bacteria. The active ingredient of the present invention obtained in Production Example 2 was added to each petri dish from 200 μg to 50%.
As a result, the number of colonies that grew regardless of the presence or absence of the metabolic activation system was suddenly returned from the negative control (without the addition of the active ingredient of the present invention). Since the number of mutant colonies did not exceed twice, the mutagenicity of the active ingredient of the present invention was determined to be negative.

(3) Sensitization test (patch test on guinea pig) Hartley female guinea pig 1 weighing 330 to 400 g
The back of 0 animals was shaved, 20 μl of an acetone solution containing 5% by weight of the active ingredient of the present invention obtained in Production Example 2 was applied, and an adjuvant patch test was performed. The positive rate was 0%. The sensitizing effect of the active ingredient was determined to be extremely low.

(4) Sensitization test (patch test on human) Lanolin solution containing 2% by weight of the active ingredient of the present invention obtained in Production Examples 2 and 3 in 30 healthy men and women aged 20 to 55 years When a patch test was carried out in accordance with a conventional method using the method described above, no irritation was observed at any of the application sites in all 30 persons, and the irritation to human skin was judged to be negative.

(5) Primary skin irritation test Hartley female guinea pig 5 weighing 330 to 400 g
The back of each animal was shaved, and 20 μl of 50% aqueous ethanol solution containing 5% by weight of the active ingredient of the present invention obtained in Production Example 2 was used.
24 hours and 48 hours after the application, the condition of the skin at the application part was observed, and the formation of erythema and edema was evaluated.
As a result, the positive rate was 0%, and it was determined that the primary irritancy to the skin of the active ingredient of the present invention was extremely low.

(6) Phototoxicity test Hartley female guinea pig 5 weighing 330 to 400 g
The back of each animal was shaved, and 20 μl each of a 50% aqueous ethanol solution containing 10% of the active ingredient of the present invention obtained in Production Example 2 was applied, and then each was irradiated with a light source (light source: Toshiba FL20S · B).
(LB, 5 lamp parallel device), and was fixed at a distance of about 10 cm, and irradiated with long-wave ultraviolet light (UV-A) for 1 hour. At 24 hours and 48 hours after the irradiation, the state of the skin at the applied portion was observed, and the formation of erythema and edema was evaluated. As a result, the positive rate was 0%, and it was determined that the phototoxicity of the active ingredient of the present invention was extremely low.

As is evident from the results of the above tests, the extract of the seeds or seed shells of the plant belonging to the genus Pecan of the family Walnuts, which is the active ingredient of the present invention, has an extremely high active oxygen scavenging effect and a lipid peroxidation inhibitory effect. have. Moreover,
The components of this extract are related plants, Walnut (Juglans)
It is a component that is different from the active oxygen scavenging active component of walnut shell extract belonging to the above and has high safety.

Examples of the active oxygen scavenger of the present invention and foods, feeds and cosmetics containing the active ingredient will be described below. Example 1 Tablets The components of the following formulation were mixed by a conventional method, the particle size was adjusted through a 60-mesh wire net, and then one tablet was produced using a tableting machine.

[0061]

[Table 8]

Example 2 Capsules The components of the following formulation were mixed well by a conventional method, and the particle size was adjusted through a 60-mesh wire net, and then filled into gelatin capsules to prepare one capsule.

[0063]

[Table 9]

Example 3 Powder The following components were uniformly mixed by a conventional method to produce one powder.

[0065]

[Table 10]

Example 4 Injection The following components were mixed in a conventional manner, filtered, filled in a container, sealed, and then subjected to high-pressure steam sterilization to produce an injection ampule.

[0067]

[Table 11]

Example 5 Gel Ointment 10 g of a gel ointment was produced by uniformly mixing and dissolving the components of the following formulation by a conventional method.

[0069]

[Table 12]

Example 6 Mayonnaise A component having the following formulation was thoroughly stirred and mixed in a mixer.
While continuing to stir, slowly add the component b, and add mayonnaise 10
0 g was produced.

[0071]

[Table 13]

[0072] adding hot water extract of shells of pecan nut obtained in Example 7 Mayonnaise commercial mayonnaise in Preparation 3, O ₂ after creation and 48 hours ^- was measured scavenging activity. O ₂ of each concentration extract ^- the erasure ratio shown in Table 14.

[0073]

[Table 14]

Example 8 Candy A suitable amount of water was added to sucrose and starch syrup of the following components, and the mixture was heated to 150 ° C. under normal pressure under ordinary pressure. One was produced.

[0075]

[Table 15]

Example 9 Chewing gum The following components were kneaded in a conventional manner with a kneader to produce one chewing gum.

[0077]

[Table 16]

Example 10 Beverage The components of the following formulation were stirred and mixed uniformly by a conventional method, and the filtered filtrate was bottled and sterilized to produce one bottle of beverage.

[0079]

[Table 17]

Example 11 Beverage The hot water extract of the pecan nut shell obtained in Production Example 3 was added to a commercially available orange juice, and the mixture was prepared and 48 hours later.
O ₂ ^- was measured scavenging activity. O ₂ of each concentration extract ^- the erasure ratio shown in Table 18.

[0081]

[Table 18]

[0082] adding hot water extract of shells of pecan nut obtained in Preparation Example 3 to Example 12 soup commercial soup, O ₂ after creation and 48 hours ^- was measured scavenging activity. O ₂ of each concentration extract ^- the erasure ratio first
The results are shown in Table 9.

[0083]

[Table 19]

Example 13 Pet Food An appropriate amount of water was added to the components of the following formulation, mixed and sifted with a sifter, and then one can of pet food was produced by a conventional method.

[0085]

[Table 20]

Example 14 Toner 1 Toner 1 was prepared by uniformly mixing the following components according to a conventional method.
00 g was produced.

[0087]

[Table 21]

Example 15 Emulsion The component a of the following formulation was dissolved by heating at 75 ° C. and the component b was heated to 73 ° C., and the component b was gradually added to the component a while stirring to emulsify. Perfume was added and mixed uniformly to prepare 100 g of an emulsion.

[0089]

[Table 22]

Example 16 Emulsion To the emulsion of Example 15 (excluding the flavor), the 50% aqueous ethanol extract of the pecan nut shell obtained in Production Example 1 was added, and O _{2 was} prepared and 48 hours later. ^- was measured scavenging activity. O ₂ of each concentration extract ^- the erasure ratio shown in Table 23.

[0091]

[Table 23]

Example 17 Cream Component a of the following formulation was heated and melted at 73 ° C. and component b was heated and melted at 75 ° C., and component b was gradually added to component a while stirring to emulsify and cooled to 55 ° C. By the way, the active ingredient and the flavor of the present invention were added, and the mixture was uniformly mixed with a homogenizer to produce 100 g of a cream.

[0093]

[Table 24]

Example 18 Bath Agent The components of the following formulation were mixed by a conventional method to produce 100 g of a bath agent.

[0095]

[Table 25]

[0096]

According to the present invention, a highly safe active ingredient having an extremely high active oxygen scavenging effect and a lipid peroxidation inhibitory effect from a specific plant seed or seed shell can be easily obtained. The composition containing the active ingredient as an active ingredient can improve and prevent various diseases caused by active oxygen and lipid peroxide and various symptoms of cosmetic disorders.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification code FI Theme coat ゛ (Reference) A23L 2/38 A61K 7/00 K 4B047 A61K 7/00 W 4C076 7/48 4C083 7/48 7/50 4C088 7/50 9/06 G 9/06 9/08 F 9/08 9/20 D 9/20 9/48 C 9/48 A23G 3/00 101 // A23G 3/00 101 3/30 3/30 A23L 1/24 A A23L 1/24 2/00 FF term (reference) 2B005 AA05 2B150 AA01 AA05 AA06 AA08 AB20 DD31 DD42 DD57 4B014 GB06 GB07 GB13 GG18 GK12 GP01 GP27 4B017 LC03 LG05 LP01 LP03 4B018 LB01 LB08 LB09 LE03 MD57 ME06 MF06 4B047 LB09 LE03 LG40 LP01 LP02 LP07 4C076 AA07 AA12 AA13 AA30 AA37 AA56 BB01 BB31 CC21 DD28C DD38A DD41C DD50F DD67A EE08A EE31A EE32A EE32B EE38A EE42H EE53A 112A FF53A AC242 AC432 AC542 AD092 AD212 AD242 AD262 AD272 BB51 CC03 DD14 DD15 DD17 DD22 DD23 DD27 DD41 EE11 EE13 4C088 AB12 AC04 AC14 BA08 BA09 BA10 MA17 MA52 MA63 NA14 ZA89 ZC21 ZC61

Claims (3)

[Claims] 1. An active oxygen scavenger containing a seed or a seed shell extract of a plant belonging to the genus Pecan of the walnut family. 2. The extract according to claim 1, wherein the extract has been extracted with one or more mixed solvents selected from water, methanol, ethanol, n-propanol, iso-propanol and acetone. The active oxygen scavenger according to claim 1. 3. A food, feed or cosmetic containing the active oxygen scavenger according to claim 1.