JP4414711B2 - Active oxygen scavenger and method for producing the same - Google Patents

Description

  The present invention relates to an active oxygen scavenger containing an anthocyanin pigment decomposition product and a method for producing the same.

Active oxygen is oxygen in an energy state rich in reactivity, and specifically refers to OH ⁻ (hydroxy radical), O ₂ ⁻ (superoxide anion), and H ₂ O ₂ (hydrogen peroxide). It has been reported that these are involved in various reactions in vivo and can cause diseases and aging.

For example, when bacteria and viruses into the body from entering, neutrophils to the local is phagocytes, monocytes, macrophages migrate to initiate phagocytosis against invading bacteria and viruses, subsequently O ₂ ^- production of Thus, this O ₂ ⁻ acts as a bactericidal, virucidal and infected cell, and is a self-protection of the living body. However, excessive O ₂ ⁻ causes damage to normal cells and tissues, such as rheumatoid arthritis, arteriosclerosis, diabetes, hepatitis, nephritis, dermatitis, blemishes, wrinkles, eczema, buckwheat Various symptoms that cause cosmetic problems are caused.

  In recent years, in order to ameliorate and prevent the above-mentioned diseases, there has been an operation of screening substances having active oxygen scavenging activity from natural products, fermented products, or chemical products from the viewpoint of pharmaceutical, food science or cosmetic science. As a result, a significant number of active oxygen scavengers have become known.

  For example, Patent Document 1 discloses a water-soluble antioxidant derived from safflower leaves and / or stems, and Patent Document 2 discloses one or more kinds of plants selected from plums, psyllium, bengal karatachi, miracle berry, yaba santa, and lemongrass. An active oxygen scavenger and cosmetic composition containing an extract are disclosed in Patent Document 3 as an active oxygen scavenger containing an extract from a mallet and / or a yellow flower falling water lotus as an active ingredient.

  On the other hand, anthocyanin-based pigments are pigments that are abundant in plants and the like, and are used as coloring agents by utilizing their vivid colors. Anthocyanin pigments are known to have active oxygen scavenging activity and are widely used in health foods and beverages such as supplements. Anthocyanin dyes are generally unstable dyes, and particularly under alkaline conditions, the C ring is opened and decomposed, causing discoloration. In addition, although it is easily decomposed and discolored due to the influence of temperature, light, etc., it is relatively stable under acidic conditions.

Acerola, a tropical fruit of the genus Hollytranoaceae, is a plant rich in anthocyanin pigments. Since an acerola fruit is acidic, anthocyanin pigments should be stable without light or heat. However, acerola contains an extremely large amount of vitamin C in an amount of about 1,500mg per 100g of fruit in addition to anthocyanin pigments, and as a result, anthocyanin pigments are easily decomposed and faded without giving light or heat. . It has also been found that decomposition is further promoted by iron contained in a small amount in the acerola. Degradation / fading of anthocyanin pigments by vitamin C and / or metals that occur in acerola is different in mechanism from degradation / fading by pH, temperature, light, etc., but there are few examples where the mechanism has been investigated in detail, It is not known whether the active oxygen scavenging activity of anthocyanin dyes is retained after decomposition and fading.
JP 2002-38151 A JP 2003-183120 A JP 2003-246747 A

An object of the present invention is to provide an active oxygen scavenger and a method for producing the same.
An object of the present invention is to provide an active oxygen scavenger containing an anthocyanin pigment decomposition product derived from acerola and a method for producing the same.
Another object of the present invention is to provide an active oxygen scavenger containing an anthocyanin pigment decomposition product obtained by decomposing an anthocyanin pigment with vitamin C and / or a metal, and a method for producing the same.

As a result of intensive studies in order to solve the above problems, the present inventors have found that an anthocyanin-based pigment decomposition product derived from acerola has an active oxygen scavenging activity.
The present inventors have also found that a decomposition product obtained by decomposing an anthocyanin pigment with vitamin C and / or metal has active oxygen scavenging activity.
Furthermore, the present inventors have found that the active oxygen scavenging activity increases as the decomposition of these decomposition products proceeds.

  Regarding the decomposition of anthocyanin dyes, there has been known a method in which an anthocyanin dye is decomposed to produce protocuccinic acid by reacting an alkylperoxyl radical with an anthocyanin dye (Lipids, Vol. 31, No. 12 (1996), pp1259-1263). Protocatechuic acid is a strong active oxygen scavenger. Therefore, when an anthocyanin dye is decomposed by such means, it is expected that the decomposed product also retains a strong active oxygen scavenging activity.

  However, in the present invention, vitamin C and / or metal is allowed to act on the anthocyanin pigment as a means for decomposing the anthocyanin pigment, and component analysis was performed on the anthocyanin pigment degradation product of the present invention. Was found not to have been generated. From this, it is clear that the known literature and the present invention have different anthocyanin pigment decomposition mechanisms. Furthermore, it has not been found so far that the active oxygen scavenging activity increases as the decomposition of the anthocyanin pigment decomposition product proceeds.

The present invention includes the following inventions.
(1) An active oxygen scavenger which contains an anthocyanin pigment decomposition product derived from acerola as an active ingredient.
(2) A method for producing an active oxygen scavenger comprising decomposing an anthocyanin pigment derived from acerola.
(3) An active oxygen scavenger containing, as an active ingredient, a processed product obtained by processing an acerola-derived fruit juice or an extract or processed product thereof at 30 ° C to 90 ° C.
(4) An active oxygen scavenger containing an anthocyanin pigment decomposition product obtained by decomposing an anthocyanin pigment with vitamin C and / or metal.
(5) A method for producing an active oxygen scavenger comprising decomposing an anthocyanin pigment with vitamin C and / or metal.

The active oxygen scavenger of the present invention contains an anthocyanin pigment decomposition product having active oxygen scavenging activity, and is effective in preventing and treating various diseases and aging related to active oxygen.
The present invention also provides the use of an anthocyanin-based pigment decomposition product that has not been conventionally used.
Since the active oxygen scavenger derived from acerola provided in the present invention is derived from natural ingredients, there are few problems of side effects.

Active oxygen scavenger and a manufacturing method thereof The present invention contain anthocyanin dye degradation product derived from acerola, active oxygen scavenger, and a method for producing the same containing anthocyanin pigment decomposition product derived from acerola.
The production area and variety of acerola used in the present invention are not particularly limited, but examples of production areas include Okinawa and Brazil.
As the active oxygen scavenger of the present invention, the acerola fruit juice obtained by squeezing the acerola fruit can be used as it is, but the fruit may be pulverized, pulverized or the like, or an extract thereof may be used. Or you may use as a processed material.

  When used as an extract or a processed product thereof, the organic solvent used for extraction is preferably a hydrophilic organic solvent. Examples of the hydrophilic organic solvent include alcohols such as methyl alcohol, ethyl alcohol, glycerin, propylene glycol, and 1,3-butylene glycol, acetone, tetrahydrofuran, acetonitrile, 1,4-dioxane, pyridine, dimethyl sulfoxide, N, N- Known organic solvents such as dimethylformamide and acetic acid can be mentioned. It is particularly preferable to extract using methyl alcohol. The hydrophilic organic solvent, particularly methyl alcohol and ethyl alcohol, are preferably used as a mixture with water.

The extraction conditions are not particularly limited, but a preferable temperature range is 5 ° C to 90 ° C, particularly 20 ° C to 40 ° C. The extraction time is preferably about 1 hour to 10 hours, particularly about 1 hour to 2 hours, and the amount of solvent used for extraction is preferably 1 to 20 times by mass with respect to the raw material.
After extraction, the extraction residue is removed by filtration or centrifugation to obtain an extract. This extract may be concentrated as necessary.

  The acerola juice obtained as described above or an extract or processed product thereof contains anthocyanin pigment, vitamin C and iron, and anthocyanin pigment can be added to vitamin without adding vitamin C and / or metal. It can be discolored by decomposition with C and / or iron. However, vitamin C and / or metal may be added to acerola juice or an extract or processed product thereof.

  The decomposition conditions are not limited, but for example, the decomposition temperature is preferably 30 ° C to 90 ° C. The decomposition time can be appropriately changed depending on the decomposition temperature. For example, it is preferably about 10 to 50 days if the decomposition temperature is a low temperature condition of 30 ° C., and preferably about 1 to 5 hours if the decomposition temperature is a high temperature condition of 90 ° C. Particularly preferable decomposition conditions are 10 to 50 days at a low temperature of 30 to 50 ° C.

An active oxygen scavenger containing an anthocyanin pigment degradation product and a process for producing the same The present inventors have also disclosed that a degradation product obtained by decomposing a general anthocyanin pigment with vitamin C and / or metal also has an active oxygen scavenging activity. Furthermore, it was found that the active oxygen scavenging activity of this decomposed product increases as the decomposition proceeds.

Examples of the anthocyanin dye include cyanidin, malvidin, delphinidin, petunidin, peonidin, pelargonidin, and glycosides containing them as aglycones, and cyanidin or glycosides thereof are particularly preferable.
As vitamin C, a commercially available product may be used.

  The metal used for the decomposition is not particularly limited as long as it promotes the decomposition of the anthocyanin pigment, and examples thereof include metal salts such as iron and copper, and iron is particularly preferable.

  Anthocyanin pigments can be decomposed with either vitamin C or metal, but are preferably decomposed with both vitamin C and metal.

  The amount of vitamin C added is preferably 5 to 100 times, particularly 50 to 100 times that of the anthocyanin pigment. The amount of the metal added is preferably in the range of 1/10 to 1/2, particularly in the range of 1/10 to 1/4 of the anthocyanin dye.

  The conditions for the decomposition are not particularly limited, but can be carried out in the same manner as the above-described decomposition of an anthocyanin pigment derived from acerola.

  The anthocyanin pigment decomposition product of the present invention can be used as an active oxygen scavenger in the form of a mixture with vitamin C and metal, but may be subjected to a purification step for removing residual vitamin C and metal after production. Examples of the purification method include normal phase or reverse phase chromatography, ion exchange chromatography, gel filtration, and the like, and the use of reverse phase chromatography is particularly preferable. A combination of these methods can also be used.

Use The active oxygen scavenger of the present invention can be formulated by combining acerola juice, extract or purified product, or anthocyanin pigment decomposition product with a known pharmaceutical carrier. The dosage form is not particularly limited and is appropriately selected as necessary. In general, tablets, capsules, granules, fine granules, powders, solutions, syrups, suspensions, emulsions, elixirs, etc. It is used as oral preparations or parenteral preparations such as injections, drops, suppositories, inhalants, transdermal absorbents, transmucosal absorbents, patches, ointments and the like. The amount of the active oxygen scavenger added according to the present invention varies depending on various conditions such as the kind of the object to be added and the form of use, but it is usually preferable to use it in the range of 0.01% by mass to 10% by mass with respect to the entire object to be added. .

  The dose of the active oxygen scavenger of the present invention varies depending on the patient's age, body weight, degree of disease, and administration route, but in oral administration, it is usually 10 mg to 3000 mg as acerola extract dry powder, The frequency of administration is usually 1 to 3 times a day for oral administration.

  Oral preparations are produced according to a conventional method using excipients such as starch, lactose, sucrose, mannitol, carboxymethylcellulose, corn starch, and inorganic salts.

  In this type of preparation, a binder, a disintegrant, a surfactant, a lubricant, a fluidity promoter, a corrigent, a colorant, a fragrance and the like can be appropriately used in addition to the above-mentioned excipients.

  Specific examples of the binder include crystalline cellulose, crystalline cellulose / carmellose sodium, methylcellulose, hydroxypropylcellulose, low-substituted hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose phthalate, hydroxypropylmethylcellulose acetate succinate, carmellose sodium , Ethylcellulose, carboxymethylethylcellulose, hydroxyethylcellulose, wheat starch, rice starch, corn starch, potato starch, dextrin, pregelatinized starch, partially pregelatinized starch, hydroxypropyl starch, pullulan, polyvinylpyrrolidone, aminoalkyl methacrylate copolymer E, aminoalkyl METAKU Rate copolymer RS, methacrylic acid copolymer L, methacrylic acid copolymer, polyvinyl acetal diethylamino acetate, polyvinyl alcohol, gum arabic, gum arabic powder, agar, gelatin, white shellac, tragacanth, purified sucrose, macrogol.

  Specific examples of the disintegrant include crystalline cellulose, methylcellulose, low-substituted hydroxypropylcellulose, carmellose, carmellose calcium, carmellose sodium, croscarmellose sodium, wheat starch, rice starch, corn starch, potato starch, and partially pregelatinized. Starch, hydroxypropyl starch, sodium carboxymethyl starch, tragacanth can be mentioned.

  Specific examples of surfactants include soybean lecithin, sucrose fatty acid ester, polyoxyl stearate, polyoxyethylene hydrogenated castor oil, polyoxyethylene polyoxypropylene glycol, sorbitan sesquioleate, sorbitan trioleate, sorbitan monostearate Sorbitan monopalmitate, sorbitan monolaurate, polysorbate, glyceryl monostearate, sodium lauryl sulfate, lauromacrogol.

  Specific examples of lubricants include wheat starch, rice starch, corn starch, stearic acid, calcium stearate, magnesium stearate, hydrous silicon dioxide, light anhydrous silicic acid, synthetic aluminum silicate, dry aluminum hydroxide gel, talc, Examples thereof include magnesium aluminate metasilicate, calcium hydrogen phosphate, anhydrous calcium hydrogen phosphate, sucrose fatty acid ester, waxes, hydrogenated vegetable oil, and polyethylene glycol.

  Specific examples of the fluidity promoter include hydrous silicon dioxide, light anhydrous silicic acid, dry aluminum hydroxide gel, synthetic aluminum silicate, and magnesium silicate.

  Moreover, the active oxygen scavenger of this invention may contain a flavoring agent and a coloring agent, when administering as a liquid agent, a syrup agent, a suspension agent, an emulsion, and an elixir.

  The active oxygen scavenger of the present invention can also be added to skin care cosmetics such as skin lotions, emulsions, creams, packs, cleaning agents, makeup cosmetics such as lipsticks and foundations, hair cosmetics, etc. according to a conventional method. it can.

  The active oxygen scavenger of the present invention can be further added to foods, rice cakes, troches, jams, chewing gums, beverages and the like according to conventional methods.

  Moreover, the active oxygen scavenger of this invention can also be used as it is as an extract dry powder. Furthermore, it may be a so-called food for specified health use (for example, food for preventing arteriosclerosis).

  Metals such as acerola, anthocyanin pigments, vitamin C and iron used in the production of the active oxygen scavenger of the present invention have been used in cosmetics, foods, etc., and safety has been established.

EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples. However, the scope of the present invention is not limited to these examples.
Example 1
The acerola juice was discolored by storage, and the active oxygen scavenging activity of the pigment degradation product was evaluated.
[Sample preparation]
The acerola was squeezed with gauze, and pectinase was allowed to act on the juice obtained after centrifugal filtration for 2 hours to eliminate the viscosity. The obtained liquid was sterilized by filter and incubated aseptically at 40 ° C. for about 1 month. By this treatment, the red color of the acerola juice gradually faded, and after 5 days it completely faded. After 0, 1, 3, 7, 14, 21, and 35 days of incubation, the bleached fruit juice was subjected to a water-soluble component on a C18 cartridge column (BAKERBOND spe (registered trademark) C18 disposable column), thoroughly washed with distilled water, The C18 column adsorption fraction was eluted with 0.2% TFA / methanol solution. The obtained eluate was concentrated with an evaporator and redissolved with distilled water in the same amount as the bleached fruit juice applied to the column. The active oxygen scavenging activity was evaluated for the sample obtained by the above operation and a sample obtained by diluting it twice or 4 times.

[Evaluation of active oxygen scavenging activity]
The active oxygen scavenging activity was evaluated based on the scavenging activity of superoxide anion radicals by a xanthine-xanthine oxidase reaction system, and ESR was used to detect the radicals.

  DMPO 15 μl, 50 mM phosphate buffer (pH 7.4) 50 μl, sample 50 μl, 2 mM hypoxanthine / phosphate buffer solution 50 μl, 0.4 unit / ml xanthine oxidase (XOD) / phosphate buffer 50 μl, and after addition of XOD ESR measurement was performed after 45 seconds.

The ESR device was ESR (JES-FR30) manufactured by JEOL Ltd., and spectrum analysis by ESR was performed under the following conditions:
Magnetic field sweep width: 335.9 ± 5mT, magnetic field modulation: 0.1mT, gain: 100, sweep time 2 minutes, response time: 0.1 seconds, measurement temperature: room temperature.

  The measurement results are as shown in FIG. As shown in FIG. 1, the active oxygen scavenging activity tended to increase as the fruit juice decomposition progressed.

Example 2
From Example 1, the acerola juice showed a tendency to increase the active oxygen scavenging activity as the decomposition progressed. In this system, the effect of components other than the anthocyanin dye is also considered, so the anthocyanin dye is purified from the acerola. The same experiment was conducted in a simple system using a purified dye product.

The final concentrations of anthocyanin pigment, vitamin C, and iron chloride purified from acerola are adjusted with 0.1M acetate buffer (pH 3.0) to 0.2, 10, and 0.02 mg / ml, respectively, according to the acerola fruit content. Incubated at 40 ° C. for about 1 month. Thereafter, the prepared solution after 0, 7, 14, 21, 28, and 35 days of incubation was applied to a C18 cartridge column, washed thoroughly with distilled water, and then the C18 column adsorbed fraction was eluted with a 0.2% TFA / methanol solution. The obtained eluate was concentrated with an evaporator and then redissolved with the same amount of distilled water as the prepared solution used for the column. The active oxygen scavenging activity was evaluated for the sample obtained by the above operation and the sample diluted 2 times, 4 times, 8 times and 16 times.
The evaluation method of the active oxygen scavenging activity is the same as in Example 1.

  The measurement results are as shown in FIG. From FIG. 2, as in the case of acerola juice, the active oxygen scavenging activity tended to increase as the decomposition of the pigment progressed. This experiment is conducted in a simple system, and the components adsorbed on the C18 column are limited to anthocyanin pigment decomposition products. Actually, vitamin C content of this adsorbed component was examined, but no vitamin C was detected. Therefore, it was found that the active oxygen scavenging activity of the anthocyanin dyes in acerola increases as the decomposition proceeds.

Example 3
In the anthocyanin pigments in acerola juice, it was found that the active oxygen scavenging activity increased as the decomposition progressed, but it was investigated whether the same results could be obtained with general anthocyanin pigments.

  Standard cyanidin chloride dissolved in 0.1M acetic acid buffer (pH 3.0) to a concentration of 0.075mg / ml, vitamin C added at 10mg / ml, iron chloride added at 0.02mg / ml, incubated at 40 ° C And faded. Thereafter, vitamin C was removed with a C18 cartridge column, and the anthocyanin pigment decomposition product was eluted with ethanol. The obtained eluate was concentrated with an evaporator, and then redissolved with distilled water to a concentration of 0.25 mg / ml. The active oxygen scavenging activity of this sample was evaluated by the DPPH method as follows.

[Evaluation of active oxygen scavenging activity]
Evaluation was made by the DPPH method. 0.5 ml of a sample was mixed with 2.5 ml of 0.1 M phosphate buffer (pH 6.5), 1.0 ml of 99% ethanol, and 1.0 ml of 5 × 10 ⁻⁴ M DPPH-ethanol solution and left at room temperature. Absorbance at 525 nm was measured over time to evaluate radical scavenging activity.
The measurement results are as shown in FIG. From FIG. 3, it was found that the activity after fading was stronger than that before fading.

It is the figure which showed the time-dependent change of the active oxygen scavenging activity of the anthocyanin pigment decomposition product produced | generated by discoloration of acerola fruit juice. It is the figure which showed the time-dependent change of the active oxygen scavenging activity of the anthocyanin type pigment decomposition product produced | generated when the anthocyanin type pigment refine | purified from acerola was discolored in coexistence with vitamin C and iron. It is the figure which showed the time-dependent change of DPPH radical scavenging activity of the anthocyanin-type pigment decomposition product produced when the cyanidin chloride of a sample is faded in the presence of vitamin C and iron.

Claims (1)

A method for producing an active oxygen scavenger comprising decomposing an anthocyanin pigment with vitamin C and / or a metal, wherein the decomposing conditions are aseptic conditions of 30 to 50 ° C for 10 to 50 days .

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