JP2006232807A - Antioxidant containing horseradish extract - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an antioxidant which has excellent safety and solves a problem that it is difficult to industrially stably obtain conventional natural substance-derived antioxidants in large amounts and a problem that synthetic antioxidants are shunned by consumers from the aspect of safety. <P>SOLUTION: An extract having an anti-oxidizing action is obtained from a horseradish (Cochlearia armoracia L.) which is a cruciferous perennial plant. The antioxidant is characterized by containing the extract. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a drug excellent in antioxidant action. More specifically, regarding an antioxidant containing an extract obtained from horseradish (cochlearia armoracia L.), an antioxidant effect, in particular, an excellent antioxidant effect such as an effect of improving the storage stability of the product, etc. The present invention relates to an antioxidant having excellent in vivo antioxidant effects such as various disease preventing effects and anti-aging effects.

Living organisms efficiently obtain energy necessary for survival by using oxygen. However, reactive oxygen species are generated as intermediates in the process of oxygen being converted to water in such energy metabolism. In general, as the reactive oxygen species, superoxide released by macrophage stimulation or the like, hydroxy radicals generated by radiation exposure, or the like are known. These reactive oxygen species are generated due to external triggers such as excessive radiation and ultraviolet radiation, intake of chemicals and tobacco, and internal factors such as ischemia reperfusion, inflammation, stress, and aging.

In general, reactive oxygen species produced in excess in the living body are generally highly chemically reactive, and easily react with adjacent components such as lipids, nucleic acids, and proteins in the living body, leading to various diseases. Cause disability. Superoxide, a kind of reactive oxygen species, has been shown to be deeply related to various diseases. For example, LDL in arteries is oxidized by superoxide to form foam cells, causing arteriosclerosis. Is generated. Moreover, the production of hydroxy radicals brought about by irradiation of radiation gives serious damages such as carcinogenesis to the living body. (Refer nonpatent literature 1).

As the toxicity of such reactive oxygen species to the living body becomes clear, antioxidants such as reactive oxygen species-eliminating substances having the activity of efficiently eliminating these active oxygen species are included in the living body or in foods, pharmaceuticals, agricultural chemicals, etc. It is useful as a protective agent against oxidative degradation of ingredients, and is expected to be used practically in the food industry, in particular, processed fishery products, health foods, and nutritional foods, as well as in the pharmaceutical / agrochemical and cosmetic fields.

  Antioxidants can be used as whitening agents in the cosmetic field. Pigmentation on the skin such as spots and buckwheat is caused by vigorous production of melanin by increasing the tyrosinase activity in melanoma cells that synthesize melanin when exposed to ultraviolet rays. This melanin is widely distributed in the animal and plant kingdoms, but in vertebrates, tyrosine is oxidized by tyrosinase in the cytoplasmic granule melanosome in melanocytes, and dopa and dopaquinone are biosynthesized. Is known to be indolequinone and the like, and melanin is biosynthesized through a complicated pathway (see Non-Patent Document 2). It is known that a whitening effect can be obtained by inhibiting an oxidation reaction by tyrosinase or an oxidation reaction in a melanin biosynthesis process with an antioxidant (see Non-Patent Document 2).

In recent years, consumers have become more aware of the safety issues of food additives made of chemically synthesized products as well as antioxidants. For example, chemicals such as BHA (butylhydroxyanisole) and BHT (butylhydroxytoluene) Synthetic antioxidants tend to avoid their use. Further, many other antioxidants are oil-soluble (water-insoluble) such as vegetable oil-derived tocopherols, and are often restricted in actual use.

Under such circumstances, various antioxidants have been extracted mainly from raw materials derived from natural products, and their application has been studied. For example, superoxide dismutase (SOD), which is an enzyme protein, has superoxide scavenging activity, and mannitol, tryptophan, formic acid, and the like have hydroxyradical scavenging activity (Non-patent Document 3).

However, since SOD is an enzyme protein, its stability against heat and the like is poor, and when administered orally, most of the administered enzyme is digested and excreted, and its effective power is extremely low. At present, there are few practical hydroxy radical scavengers that can efficiently scavenge hydroxy radicals. Therefore, it is difficult to obtain these antioxidants industrially in large quantities and stably. In addition, the above-mentioned SOD usually has an erasing effect only on superoxide and has no effect on hydroxy radicals. Similarly, mannitol cannot eliminate superoxide.

  Antioxidants from plants are also being searched for, but if an inadvertent plant is selected to obtain an extract, there is a risk of contamination with alkaloids that cause the majority of plant toxins. Alkaloid is an alkaline molecule containing nitrogen, which is similar to a neurohormone, so it enters the end of nerve fibers and defeats the action of nerves and exerts toxicity. Therefore, care must be taken in selecting plants and extracting active ingredients.

Halliwell B.M. and Gutteridge M.M. C. Biochem. J. et al. 219, 1-14, (1984)) Development of functional cosmetics, CMC Publishing, 2000 SOD and reactive oxygen species regulator-its pharmacological action and clinical application, pp. 224-228, Nippon Medical Museum, 1989

In the prior art, it is difficult to industrially obtain a large amount of natural-derived antioxidants stably, and synthetic products have a problem of being avoided by consumers in terms of safety. Accordingly, an object of the present invention is to provide an antioxidant excellent in safety.

The present inventors have surprisingly found that an extract of horseradish, which is a food plant, has an antioxidant action, and further, an antioxidant containing such an extract as an active ingredient is, for example, cream or The present invention has been completed by finding that it is effective as an external preparation for skin such as lotion.

In addition, the present inventors have found that an extract of horseradish containing the active ingredient can be obtained not only from the rootstock of horseradish but also from leaves and stems of horseradish or various residues. Horseradish usually uses only rhizome as a food, and rhizome residues obtained by extracting certain enzymes as well as leaves and stems are often discarded as they are, and the use of waste wood is expected by using the present invention. Is done.

As a result of intensive studies, the present inventors have found that the above problems can be solved by the following means, and have reached the present invention. That is, the present invention relates to an antioxidant excellent in safety.
1. An antioxidant comprising a horseradish extract as an active ingredient.
2. 1. An antioxidant comprising obtaining an extract containing an active ingredient from one or a mixture of two or more selected from the group consisting of horseradish leaf, horseradish stem and / or horseradish rhizome .
3. One or two antioxidants characterized in that the horseradish is horseradish derived from Cochlearia armoracia L.
4). One or two antioxidants, wherein the action of the active ingredient is an antioxidant action.
5. One or two antioxidants characterized in that polyphenols are contained in the horseradish extract.
6). 1 or 2 antioxidant characterized by including ascorbic acid and / or an ascorbic acid derivative, and / or arbutin and / or an arbutin derivative in addition to the extract of a horseradish.
7). An antioxidant comprising horseradish extract as an active ingredient.
8). The horseradish extract is one or a mixture of two or more selected from the group consisting of horseradish leaf extract, horseradish stem extract and / or horseradish rhizome extract 7 antioxidants characterized.
9. 7 or 8 antioxidant, characterized in that the horseradish is horseradish from Cochlearia armoracia L.
10. Furthermore, ascorbic acid and / or an ascorbic acid derivative, and / or arbutin and / or an arbutin derivative, any one of 7-9 characterized by the above-mentioned.

  INDUSTRIAL APPLICABILITY According to the present invention, it is possible to stably provide an antioxidant containing a natural extract having a powerful superoxide scavenging function, an active oxygen scavenging function such as hydroxy radical scavenging, a tyrosinase inhibiting action and a melanin production inhibiting action. . The anti-oxidant of the present invention has a strong anti-oxidation effect in a trace amount. By using the anti-oxidant for the human body, other foods and drinks, pharmaceuticals, cosmetics, whitening agents, fertilizers, feeds, skin external preparations and the like. In addition, it is possible to obtain an excellent antioxidant effect such as an antioxidant effect, in particular, an effect of improving the storage stability of the product, and an excellent in vivo antioxidant effect such as various disease preventing effects and anti-aging effects. In addition, the use of biomass is expected to make a significant contribution to the biomass industry.

As a result of diligent studies to achieve the above-mentioned problems, the extract obtained from horseradish has an antioxidant effect, particularly an excellent antioxidant effect such as an effect of improving the storage stability of the product, and various diseases. It has been found that it has an excellent in vivo antioxidant effect such as a preventive effect and an anti-aging effect and a whitening effect, and the present invention has been completed. That is, the present invention relates to an antioxidant containing an extract obtained from horseradish as an active ingredient.

The horseradish used in the present invention is known to be used as a spicy seasoning for roast beef by grinding down its rhizomes, but this horseradish extract is effective as an antioxidant. Not known at all.

  In the extraction of active ingredients from horseradish, extraction may be performed from the whole plant body. In addition, the extraction operation may be performed by dividing the plant body into rhizomes, stems, and leaves, or may be performed in combination with each other. In addition, it is generally known that a certain enzyme is extracted from horseradish rhizome, but it may be extracted from a residue obtained after the enzyme is extracted and discarded. In the present invention, it has been found that among the wasabi roots, stems, and leaves used, especially the leaves have active ingredients, and therefore it is preferable to perform the extraction operation from the leaves. Horseradish is known to be used as a spice for roast beef by grinding its rhizomes, but its leaves are discarded, and its effective use is expected. By using the leaves of horseradish, it can be expected to contribute greatly to producers of horseradish and industries in the biomass field.

  The horseradish plant may be extracted from any of a raw state, a frozen state, and a lyophilized state. Preferably, the extraction operation is performed from the raw state. When extracting from raw or frozen horseradish, it may be crushed first. What is necessary is just to crush using a mixer, a homogenizer, a mortar, etc. for crushing. The freeze-dried product may be crushed into a powder after the freeze-drying operation.

The crushed plant body is extracted using a solvent. As extraction methods, a method of extracting by impregnating at room temperature, in a cooled or heated state, a method of extracting using a distillation method such as steam distillation, a pressing method of obtaining an extract by pressing, a supercritical fluid or a subcritical fluid Extraction methods may be used, and these methods may be used alone or in combination of two or more. Preferably, 5 to 100 ml of solvent is used per 1 g of finely cut raw material, and it is desirable to carry out at room temperature or under heating for 1 hour to 1 month, preferably 1 to 5 days.

The solvent used for the extraction is not particularly limited as long as the active ingredient can be effectively extracted, but water, an organic solvent miscible with water such as a lower alcohol such as ethanol, or the like thereof. A mixed solution or an organic solvent immiscible with water, such as ethyl acetate, may be used. Other examples include monohydric alcohols such as methanol, isopropanol, isobutanol, n-hexanol, methyl amyl alcohol, 2-ethyl butanol, n-octyl alcohol, glycerin, ethylene glycol, ethylene glycol monomethyl ether, Polypropylene glycol, propylene glycol monomethyl ether, propylene glycol monoethyl ether, triethylene glycol, 1,3-butylene glycol, hexylene glycol and other polyhydric alcohols or derivatives thereof, acetone, methyl ethyl ketone, methyl isobutyl ketone, methyl-n-propyl Ketones such as ketones, esters such as isopropyl acetate, ethers such as ethyl ether, isopropyl ether and n-butyl ether, squalane, petrolatum, para Examples include hydrocarbons such as oil wax, paraffin oil, olive oil, wheat germ oil, rice oil, sesame oil, macadamian nut oil, almond oil, coconut oil and other animal oils such as beef tallow, pork fat, whale oil The Moreover, the polar solvent which added inorganic salts, such as phosphate buffered saline, and the solvent which added surfactant can also be used, It does not specifically limit. Considering safety in use for food and drink, application to the human body including application to the skin, etc., extraction with water and / or alcohol is particularly preferable.

  Moreover, you may perform extraction operation in steps. For example, perform extraction with an organic solvent that is miscible with water, such as lower alcohols such as ethanol, concentrate with an evaporator, etc., add an organic solvent that is immiscible with water, such as ethyl acetate and water, and separate the organic layer from the aqueous layer. You may do it. In this case, components having a tyrosinase inhibitory action, a melanin production inhibitory action, and an antioxidant action are extracted in both the organic layer and the aqueous layer, and may be used for different purposes. For example, the components of the aqueous layer can be expected to be used when an anti-oxidant that has an affinity for water is desired, such as facial cleansing applications, skin lotion applications, and use applications. Although it can be expected to be utilized when an antioxidant such as an emulsion that requires affinity for oil is desired, it is not limited thereto.

The extract of horseradish thus obtained can be used as it is, but may be used after adding purification operations such as deodorization and decolorization within the range not losing the effect. Moreover, in order to increase the concentration of the active ingredient in the extract, the obtained extract may be further subjected to means such as concentration, liquid-liquid distribution, adsorption chromatography, normal phase or reverse phase chromatography, if desired. Is possible. These extracts, purified products or fractions can be made into a dry product by removing the solvent from them, and further used in a form solubilized in a solvent such as alcohol, or in the form of an emulsion. it can.

The horseradish extract may be used in combination with other tyrosinase inhibitory action, melanin production inhibitory action and / or antioxidant action. Synergistic effects can be expected when used in combination with other drugs. When used in combination, it is preferable to use a highly safe drug in combination. More preferably, a naturally occurring drug or a drug imitating a naturally derived drug is used in combination. For example, ascorbic acid and the like are naturally present and are also used for food applications, so they are relatively easy to use together. Examples of L-ascorbic acid and salts or derivatives thereof that may be used in the present invention include monoalkyl ascorbates such as L-ascorbic acid monostearate, L-ascorbic acid monopalmitate, and L-ascorbic acid monooleate. Or monoalkenyl esters, ascorbic acid monoester derivatives such as L-ascorbic acid monophosphate, L-ascorbic acid-2-sulfate, L-ascorbic acid distearate, L-ascorbic acid dipalmitate, L-ascorbic acid dioleate L-ascorbic acid trialkyls such as L-ascorbic acid dialkyl or dialkenyl ester derivatives, L-ascorbic acid tristearate, L-ascorbic acid tripalmitate, L-ascorbic acid trioleate Or tri alkenyl ester derivatives,

Examples include L-ascorbic acid triester derivatives such as L-ascorbic acid triphosphate. Of these L-ascorbic acid and salts thereof or derivatives thereof, particularly preferred are L-ascorbic acid, L-ascorbic acid phosphate and salts thereof. Since arbutin derived from bilberry is also natural, it can be used in combination since it is considered relatively safe. You may use together with what imitated the structure of arbutin, or its derivative (s).

  The horseradish extract in the present invention can be used as an antioxidant as it is, and further components used in ordinary cosmetics, quasi drugs, such as oily components, surfactants, ultraviolet absorbers, lower alcohols Antiseptics, bactericides, colorants, powders, fragrances, water-soluble polymers, buffer materials, etc. can be combined with the dosage form as appropriate and blended as appropriate without compromising the effects of the present invention. is there. In this case, the extract is desirably blended in the total composition of the antioxidant in an amount of 0.0001 to 20% by weight, particularly 0.01 to 10% by weight, as the dry solid content of the plant used for extraction.

The dosage form of the external preparation for skin of the present invention is arbitrary, and can be provided as a solubilizing system such as lotion, an emulsifying system such as cream or emulsion, or a dispersing system such as calamine lotion, or a propellant. It is also possible to take aerosol dosage forms filled together.

In addition, the form of the antioxidant of the present invention includes powders, capsules (hard capsules, soft capsules, etc.), granules (coated products, pills, troches, liquids, or pharmaceuticals thereof. Acceptable sustained-release preparations, etc.).

These preparations may be used together with pharmacologically acceptable bases, carriers, excipients, disintegrants, lubricants, coloring agents, etc., in accordance with known pharmaceutical manufacturing methods.

Examples of carriers and excipients used in these preparations include lactose, glucose, sucrose, mannitol, potato starch, corn starch, calcium carbonate, calcium phosphate, calcium sulfate, crystalline cellulose, licorice powder, and gentian powder.

Examples of the binder used in these preparations include starch, tragacanth gum, gelatin, syrup, polyvinyl alcohol, polyvinyl ether, polyvinyl pyrrolidone, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, carboxymethyl cellulose and the like.

Examples of the disintegrant used in these preparations include starch, agar, gelatin powder, sodium carboxymethylcellulose, calcium carboxymethylcellulose, crystalline cellulose, calcium carbonate, sodium bicarbonate, sodium alginate and the like.

Examples of lubricants used in these preparations include magnesium stearate, talc, hydrogenated vegetable oil, macrogol and the like.

As the colorant used in these preparations, those allowed to be added to pharmaceuticals can be used.

When preparing tablets and granules, sucrose, gelatin, hydroxypropylcellulose, purified shellac, gelatin, glycerin, sorbitol, ethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, cellulose phthalate acetate, It may be coated with hydroxypropylmethylcellulose phthalate, methyl methacrylate, methacrylic acid polymer or the like, or may be coated with two or more layers. Furthermore, capsules of substances such as ethyl cellulose and gelatin may be used.

Examples of the form of the external preparation include solid, semi-solid, semi-solid, or liquid preparations for transdermal administration or transmucosal administration such as buccal or nasal administration. Examples of liquid preparations include emulsions such as pharmaceutically acceptable emulsions or lotions, tinctures for external use, and solutions for transmucosal administration. This preparation contains, for example, ethanol, oil, emulsifier and the like as a commonly used diluent. Examples of semi-solid preparations include ointments such as oily ointments and hydrophilic ointments. This preparation contains, for example, water, petrolatum, polyethylene glycol, an oil component, a surfactant and the like as a commonly used base or carrier. Examples of semi-solid or solid preparations include patches for transdermal or transmucosal (oral, nasal) administration such as plasters (rubber plaster, plaster, etc.), films, tapes, or poultices. Can be mentioned. This preparation is usually used as a base or carrier, for example, rubber polymers such as natural rubber, butadiene rubber, synthetic rubber such as SBR, SIS, mudizing agents such as gelatin, kaolin, zinc oxide, sodium carboxymethylcellulose, Contains hydrophilic polymer such as sodium polyacrylate, acrylic resin, tackifier such as liquid paraffin, water, other oils, and surfactant. These preparations may further contain additives such as stabilizers, solubilizers, and percutaneous absorption accelerators, or additives such as fragrances and preservatives.

The antioxidant of the present invention can be blended into oral and parenteral products according to conventional methods, and is used in various fields such as foods and drinks, external preparations for skin, pharmaceuticals, quasi drugs, feeds, feeds, agricultural chemicals and the like. Can be used. For example, when blended with food and drink, food and drink having an in vivo antioxidant effect and antioxidant effect can be provided. For example, this food and drink can be expected to have an antioxidant effect such as flavor deterioration and discoloration suppression, and can also be expected to have an effect such as prevention of lifestyle-related diseases. From the effect of preventing lifestyle-related diseases, it can be expected to be used as health food, nutritional food, and the like. Furthermore, the functional food and drink which exhibits the further effect can be provided by adjusting the conditions, such as concentration of this extract, by continually ingesting the food and drink containing this. In addition, it can be used for livestock and fish feeds and feeds. Especially, many fish feeds that require water are prone to oxidative degradation, and they can be easily contacted with water because they are dropped in water. However, the feed etc. which improved these can be provided. In addition, some fish for viewing and aquaculture that are difficult to breed also exist in some cases, but the feed using the antioxidant of the present invention has effects such as in vivo antioxidant effect, It is possible to provide foods and the like that contribute to the improvement of these breeding and survival rates.

In the present invention, it has been found that about 0.1 to 10 mg of polyphenol is contained in an extract extracted from 1 g of fresh heavy leaves in a horseradish extract. Polyphenol is a general term for many substances, and the center of these is a series of substances called flavonoids with similar chemical structures. Flavonoids include flavanols (catechins), flavones, isoflavones, flavonols, flavans, flavanones, chalcones, anthocyanins. In addition to flavonoids, there are phenylpropanoids, lignans, coumarins, diallylpeptanoids including curcumins, capsaicins, and the like. There are many other polyphenols.

These polyphenols are known to have antioxidation, antimutagenicity, anticancer, antihypertensive, blood glucose, antiallergic, antibacterial and antiviral effects. In particular, the antioxidant action of polyphenols has attracted attention by reducing active oxygen. These are said to be related to a plurality of hydroxyl groups (—OH), which is a structure common to all polyphenols. It is said that this part is very easy to oxidize, and polyphenols are oxidized before other coexisting substances, so that the oxidation of other substances is delayed. Further, since the ease of reaction varies depending on the type of polyphenol, it is said that it is not always easy to reduce all active oxygen with one type of polyphenol.

In the present invention, the horseradish extract has been found to have a tyrosinase inhibitory action and a melanin production inhibitory action. As a mechanism of these actions, the involvement of polyphenols contained in horseradish is presumed. Melanin pigments are produced in melanosomes, which are organelles in pigment cells (melanocytes) present in the basal layer of the epidermis, the hair root, and the outer root sheath. In the process of melanin production, tyrosinase acts on tyrosine in pigment cells (melanocytes), tyrosin is oxidized, converted to dopa and dopaquinone, and further auto-oxidized and polymerized via dopachrome and 5,6-dihydroxyindole. Finally, it becomes a melanin pigment. The melanin thus produced is secreted into the basal cells from the dendrites of the pigment cells (melanocytes), and the basal cells divide and become spiny cells and rise to reach the stratum corneum. Furthermore, it has been reported that when this melanin is irradiated with ultraviolet rays, the existing melanin is oxidized and temporarily becomes black. It is speculated that the polyphenol contained in the horseradish extract inhibits the oxidation reaction step in the production process of these melanin pigments and exhibits a tyrosinase inhibitory action and a melanin production inhibitory action.

EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to the examples.

Melanin Production Inhibition Test Method The melanin production inhibition test was performed as follows using mouse melanoma cells. First, B16 melanoma cells cultured in a DMEM medium containing 10% (v / v) fetal bovine serum in a carbon dioxide incubator adjusted to 5% (v / v) carbon dioxide at 37 ° C. to give 1 × 10 ⁵ cells. 500 μl was seeded in a 24-well cell culture plate, and then 0.5 μl of a sample of 250 mg / ml to 0.025 mg / ml dissolved in ethanol was added to the plate. After further culturing for 4 days under the same conditions, the degree of whitening was evaluated with the naked eye under a microscope and used as an index of the ability to suppress melanin production.

Tyrosinase activity inhibition test method
Anal. Biochem., 179, 375 (1989) According to the method of 179,375 (1989), 750 μl of test solution was dissolved in 750 μl of proline solution (dissolved in 80 mM, pH 7.6 phosphate buffer) and pyrocatechol solution (dissolved in 4 mM, pH 7.6 phosphate buffer). ) 750 μl was added, and enzyme tyrosinase solution (dissolved in pH 6.8 phosphate buffer) was added to a final concentration of 20 U / ml, and pH 6.8 phosphate buffer was added so that the total amount of the reaction solution was 1 ml. The mixture was quickly mixed, and the absorbance at 525 nm was immediately started, and the measured value (D ₁ ) at the start of the reaction and the measured value (D ₂ ) at 10 seconds after the reaction were obtained. That is, D ₁ was converted to 100% inhibition, and the inhibition rate at the time of D ₂ measurement was obtained (Formula 1), which was used as an index of tyrosinase activity inhibition.

Antioxidant test method Radical scavenging activity was measured for the extract. That is, DPPH which is a stable radical is prepared in ethanol to 0.5M, 0.5 ml is dispensed into a test tube, and 0.4 ml of 0.1M Tris-HCl (pH 7.4) buffer is added to this buffer. The extract solution prepared at a concentration of 0.1 ml was mixed, reacted at room temperature in a dark place for 20 minutes, and then the absorbance at 517 nm was measured. Also, Trolox, which is a powerful radical scavenger, was adjusted to 100 mM with ethanol, 0.1 ml of this was added instead of the extract solution, and the absorbance of the reaction solution reacted in the same manner was measured. Furthermore, the absorbance of a blank to which 0.1 ml of 0.1 M Tris-HCl (pH 7.4) buffer was added instead of the extract solution was measured. The radical scavenging rate of the extract was determined by Equation 2.

Measuring method of polyphenol content The total polyphenol content contained in the extract was measured using a polyphenol measuring device PA-20 (manufactured by Toyobo Engineering Co., Ltd.). The method was performed according to the PA-20 manual. 50 μl of sample was used.

Preparation of horseradish (leaves and rhizome) extracts 100 g of horseradish leaves and rhizomes were cut into small pieces, added with 1000 ml of ethanol, extracted at room temperature for 24 hours, and the extract was filtered. The residue was further re-extracted in the same manner by adding 200 ml of ethanol. The extract was collected and concentrated under reduced pressure. After distilling off the solvent, 20 ml of water and 80 ml of ethyl acetate were added, and this was transferred to a separatory funnel, shaken vigorously, and allowed to stand to separate into two layers. The ethyl acetate layer was collected. This was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 1.367 g of horseradish leaf extract and 0.2 g of rhizome extract.

Example 1 Melanin production inhibition test The melanin production inhibition was measured at various concentrations for the horseradish leaf extract by the melanin production inhibition test method described above. The results are shown in Table 1. The extract of the present invention exhibited a ++ (high whitening degree) melanin production inhibitory effect sufficient for practical use within a final concentration range of 25 μg / ml or more. This indicates that the extract of the present invention is a useful antioxidant component having both an excellent antioxidant effect and high safety.

Example 2 Inhibition of tyrosinase activity The horseradish leaf extract and the standard arbutin (synthetic product) were examined for tyrosinase enzyme inhibition activity according to the tyrosinase activity inhibition test method described above. The results are shown in Table 2.

From the results shown in Table 2, the antioxidant of the present invention consisting of arbutin and natural horseradish leaf extract inhibited tyrosinase enzyme activity at almost the same concentration.

Example 3 Antioxidant action The horseradish leaf extract was subjected to an antioxidant test according to the antioxidant test method described above. The results of the antioxidant test are shown in FIG. The leaf extract showed stronger antioxidant effect than arbutin. A slight antioxidant activity was also observed in the rhizome extract.

Example 4 Measurement of polyphenol content Using polyphenol measuring device PA-20 (manufactured by Toyobo Engineering Co., Ltd.), the total polyphenol content contained in the extract was measured. It was confirmed that about 2 mg of polyphenol was contained therein.

Example 5 Production of cream A cream having the following composition was produced by a conventional method.
The obtained cream was excellent in whitening effect and could improve pigmentation diseases such as spots and freckles.

(Composition) (wt%)
Stearic acid 2.5
Stearyl alcohol 6.0
Reduced lanolin 2.5
Squalene 6.0
Octyldecanol 6.5
Polyethylene seglycol 4.0
Fragrance 0.1
Preservative, antioxidant Suitable amount Propylene glycol 4.5
Horseradish extract 1.0
Total amount of purified water is 100

Example 6 Production of lotion A lotion having the following composition was produced by a conventional method.
The obtained lotion was excellent in whitening effect, and could improve pigmentation such as spots and freckles.

(Composition) (wt%)
Stearic acid 0.5
Cetanol 2.0
Vaseline 2.5
Lanolin alcohol 2.5
Liquid paraffin 9.0
Polyethylene seglycol 2.5
Fragrance 0.1
Glycerin 3.5
Propylene glycol 4.5
Triethanolamine 1.0
Horseradish extract 2.0
Total amount of purified water is 100

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to stably provide an antioxidant containing a natural extract having a powerful superoxide scavenging function, an active oxygen scavenging function such as hydroxy radical scavenging, a tyrosinase inhibiting action and a melanin production inhibiting action. . The anti-oxidant of the present invention has a strong anti-oxidation effect in a trace amount. By using the anti-oxidant for the human body, other foods and drinks, pharmaceuticals, cosmetics, whitening agents, fertilizers, feeds, skin external preparations and the like. In addition, it is possible to obtain an excellent antioxidant effect such as an antioxidant effect, in particular, an effect of improving the storage stability of the product, and an excellent in vivo antioxidant effect such as various disease preventing effects and anti-aging effects. Therefore, it is expected to greatly contribute to the industry. In addition, the use of biomass is expected to make a significant contribution to the biomass industry.

Results of antioxidant test

Claims (10)

An antioxidant comprising a horseradish extract as an active ingredient. The extract containing an active ingredient is obtained from 1 type, or 2 or more types of mixtures chosen from the group which consists of a horseradish leaf, a horseradish stem, and / or a horseradish rhizome. Antioxidants.   The antioxidant according to claim 1 or 2, wherein the horseradish is horseradish derived from Cochlearia armoracia L .. The antioxidant according to claim 1 or 2, wherein the action of the active ingredient is an antioxidant action. The antioxidant according to claim 1 or 2, wherein the extract of horseradish contains polyphenol. The antioxidant according to claim 1 or 2, further comprising ascorbic acid and / or an ascorbic acid derivative, and / or arbutin and / or an arbutin derivative, in addition to the horseradish extract. An antioxidant comprising horseradish extract as an active ingredient.   The horseradish extract is one or a mixture of two or more selected from the group consisting of horseradish leaf extract, horseradish stem extract and / or horseradish rhizome extract Antioxidant according to claim 7, characterized in that   The antioxidant according to claim 7 or 8, wherein the horseradish is horseradish derived from Cochlearia armoracia L ..   The antioxidant according to any one of claims 7 to 9, further comprising ascorbic acid and / or an ascorbic acid derivative and / or arbutin and / or an arbutin derivative.

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