JP2011052028A - Material treated with acerola containing polyphenol and/or vitamin c - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a preparation exhibiting effective ameliorating effect on various diseases. <P>SOLUTION: The subject treated material of acerola containing a polyphenol and/or vitamin C is useful for new pharmaceuticals, food products, drinks, cosmetics, or the like, because the material has anti-oxidation activity, arteriosclerosis prophylaxis activity, carcinogenesis suppressing activity, tumor growth suppressing activity, vascularization inhibiting activity, anti-allergic activity, anti-inflammatory activity, cholesterol reducing activity, antimicrobial activity, anti-caries activity, halitosis reducing activity, platelet aggregation inhibiting activity, anti-influenza virus activity, stomach cancer suppression activity and neutral fat absorption reducing activity. The acerola polyphenol has a vitamin C stabilizing activity and is useful for new pharmaceuticals, food products, drinks, and cosmetics, or the like by utilizing the activity. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to an acerola-treated product containing polyphenol and / or vitamin C, a production method thereof and a use thereof.

  Acerola is a tropical fruit belonging to the genus Hollytranoaceae and is native to the Caribbean Islands. Acerola fruit is known as a plant containing abundant vitamin C of about 1,500 mg or more per 100 g of fruit. Vitamin C is known to have antioxidant activity and is now used as a beverage and health food in various countries around the world.

  By the way, since acerola exhibits vivid red, it is expected that various polyphenol components are included. Polyphenol is an ingredient that has recently attracted attention as a natural ingredient that has antioxidant activity and has a preventive effect on so-called lifestyle-related diseases such as arteriosclerosis, diabetes, and cancer.

  The present inventors have included about 100 to 300 mg / 100 g of polyphenols including anthocyanin pigments and quercetin glycosides in acerola, and these acerola polyphenols exhibit a blood glucose level increase inhibitory activity and an AGE production inhibitory activity. Therefore, it has been found that it is effective for the prevention and treatment of diabetes and / or diabetic complications (Patent Document 1).

  Other common functionalities known to date for polyphenols and vitamin C include:

1. Atherosclerosis is caused by oxidation of LDL. Polyphenols are known to suppress this oxidation of LDL (Non-patent Document 1) and are effective in preventing arteriosclerosis.
2. From several animal experiments, polyphenols are known to suppress carcinogenesis by oral administration (Non-patent Document 2).
3. Polyphenols are known to have antitumor activity in green tea leaves containing catechin from experiments using sarcoma 180 as tumor cells (Non-patent Document 3).

4). Polyphenol is known to have an activity of inhibiting angiogenesis (Non-Patent Document 4).
5). Polyphenols in perilla seeds are known to have antiallergic activity (Non-patent Document 5).
6). Polyphenols are known to have anti-inflammatory activity (Non-Patent Document 6).

7). Tea catechin, which is a polyphenol, is known to have an activity of inhibiting cholesterol absorption in the small intestine to bile acid micelles (Non-patent Document 7), and is effective in suppressing an increase in plasma cholesterol.
8). Tea catechin, which is a polyphenol, is known to have an effect of inhibiting angiotensin I converting enzyme (Non-patent Document 8), and is effective as an antihypertensive agent.
9. Polyphenols contained in guava are known to have antibacterial activity (Non-patent Document 9).

Ten. Polyphenols contained in oolong tea are known to have glucosyl transferase inhibitory activity (Non-Patent Document 10) and are effective as anti-cariogenic agents.
11． Polyphenols contained in apples are known to have deodorant activity (Non-patent Document 11).
12． Polyphenols contained in red wine are known to have platelet aggregation inhibitory activity (Non-patent Document 12).

13. Polyphenols contained in twilight tea are known to have an effect of enhancing the enzyme activity of lipoprotein lipase (Non-patent Document 13) and are effective as anti-obesity agents.
14. It is known that polyphenols contained in tea have an effect of suppressing infection with influenza virus (Non-patent Document 14).
15． Quercetin and rutin, which are types of flavonoids, are known to synergistically suppress hemoglobin oxidation in the presence of vitamin C (Non-patent Document 15).
16． It is known that flavonoids contained in citrus fruits synergistically suppress LDL oxidation in the presence of vitamin C and synergistically prevent atherosclerosis (Non-patent Document 16). .
17． Vitamin C is known to show the effect of suppressing the onset of gastric cancer due to Helicobacter pylori infection (Non-patent Document 17).

  Meanwhile, hyperlipidemia is a state either or both of cholesterol and triglyceride in blood (triglyceride) becomes high, long-term intake, familial, consider the cause and aging of the high fat diet It is done. Hyperlipidemia is an etiology that causes arteriosclerosis. When diseases such as hypertension, diabetes, and obesity overlap, it leads to serious diseases such as angina pectoris, myocardial infarction, and cerebral infarction. Criteria for hyperlipidemia is that HDL to total cholesterol in the blood have been synthesized in a and liver or is 220 mg / dL or more, or triglycerides 150 mg / dL or more is less than 40 mg / dL. According to these criteria, hypercholesterolemia, hypertriglyceridemia, and mixed hyperlipidemia are diagnosed. According to the 1999 National Nutrition Survey (Ministry of Health, Labor and Welfare), the proportion of hyperlipidemic patients has increased to 59.6% in the 40s for men and 62.5% in the 50s for women. According to the 2002 National Nutrition Survey (Ministry of Health, Labor and Welfare), the proportion of lipid in the total energy intake per person per day is 25%, the upper limit of the appropriate ratio for adults in their 20s and 40s. Excessive fat has been pointed out. Conventionally known antihyperlipidemic drugs have the effect of reducing either or both cholesterol and triglycerides. Representative examples include HMG-CoA reductase inhibitors, probucol, clofibrates, nicotinic acid derivatives, EPA preparations and the like.

Japanese Patent Application No. 2003-314207

Lancet, 344, 1152, 1994 Annu. Rev. Nutr., 21, 381, 2001 Agric. Biol. Chem., 57, 1879, 1998 Proc. Natl. Acad. Sci. USA, 90, 2690, 1993 Food and Development, 32, 41, 1997 Chem. Toxic., 33, 1061, 1995 Biochim. Biophys. Acta., 1127, 141, 1992 Japanese Journal of Agricultural Chemistry, 61, 803, 1987 Biosci. Biotechnol. Biochem., 66, 1727, 2002 Appl. Env. Microbiol., 59, 968, 1993 Food and Development, 29, 43, 1994 Proceedings of the symposium, “Polyphenols, Wine and Health,” Bordeaux, France, 7, 1999 Bio. Pharm. Bull., 21, 517, 1998 Antiviral Res., 21, 289, 1993 Biomed. Pharmacotherapy, 57, 124, 2003 J. Medicinal Food, 4, 187, 2001) (J. Agric, Food Chem., 46, 1453, 1998 Gut, 50, 165, 2002

  An object of the present invention is to provide an acerola-treated product containing polyphenol and / or vitamin C, a production method thereof, and a use thereof.

  The object of the present invention is also antioxidant activity, arteriosclerosis prevention activity, carcinogenesis inhibition activity, tumor growth inhibition activity, angiogenesis inhibition activity, antiallergic activity, antiinflammatory activity, cholesterol lowering activity, antibacterial activity, anticariogenic activity, An acerola-treated product containing polyphenol and / or vitamin C having bad breath odor reducing activity, platelet aggregation inhibitory activity, anti-influenza virus activity, gastric cancer inhibitory activity, and neutral fat absorption lowering activity, and a novel preparation utilizing these activities, It is to provide food, beverages, cosmetics and the like.

  Another object of the present invention is to provide a novel preparation, food, beverage, cosmetics and the like using an acerola polyphenol having vitamin C stabilizing activity.

  The present inventors have found that acerola processed product containing polyphenols and / or vitamin C, antioxidant activity, arteriosclerosis prophylactic activity, carcinogenesis inhibition activity, tumor growth suppression activity, angiogenesis inhibitory activity, antiallergic activity, anti-inflammatory It has been found to have activity, cholesterol lowering activity, antibacterial activity, anti-cariogenic activity, bad breath odor reducing activity, platelet aggregation inhibitory activity, anti-influenza virus activity, gastric cancer inhibitory activity, and neutral fat absorption lowering activity.

  The inventors have also discovered that acerola polyphenols have vitamin C stabilizing activity.

The present invention for solving the above problems includes the following inventions.
(1) Acerola-treated product containing polyphenol and / or vitamin C.
(2) The acerola-treated product according to (1) above, wherein the acerola-treated product is an acerola juice, an acerola crushed product, an acerola extract or an acerola purified product.
(3) An antioxidant containing the acerola extract or processed product according to (1) or (2) as an active ingredient.
(4) An arteriosclerosis preventive agent containing the acerola extract or processed product according to (1) or (2) as an active ingredient.
(5) A carcinogenic inhibitor containing the acerola extract or processed product according to (1) or (2) as an active ingredient.

(6) A tumor growth inhibitor containing the acerola extract or processed product according to (1) or (2) as an active ingredient.
(7) An angiogenesis inhibitor containing the acerola extract or processed product according to (1) or (2) as an active ingredient.
(8) An antiallergic agent containing the acerola extract or processed product according to (1) or (2) as an active ingredient.
(9) An anti-inflammatory agent containing the acerola extract or processed product according to (1) or (2) as an active ingredient.
(10) A cholesterol-lowering agent containing the acerola extract or processed product according to (1) or (2) as an active ingredient.

(11) An antibacterial agent containing the acerola extract or processed product according to (1) or (2) as an active ingredient.
(12) An anti-cariogenic agent containing the acerola extract or processed product according to (1) or (2) as an active ingredient.
(13) A bad breath reducing agent containing the acerola extract or processed product according to (1) or (2) as an active ingredient.
(14) A platelet aggregation inhibitor containing the acerola extract or processed product according to (1) or (2) as an active ingredient.
(15) An anti-influenza virus agent containing the acerola extract or processed product according to (1) or (2) as an active ingredient.

(16) A gastric cancer inhibitor containing the acerola extract or processed product according to (1) or (2) as an active ingredient.
(17) An antioxidant comprising acerola polyphenol (referred to herein as acerola-derived polyphenol; the same shall apply hereinafter) and vitamin C as active ingredients.
(18) A vitamin C stabilizer containing acerola polyphenol as an active ingredient.
(19) A neutral fat absorption-lowering agent containing the acerola extract or processed product according to (1) or (2) as an active ingredient.
(20) A food containing the agent according to any one of (3), (10), (17), (18) and (19).

  The acerola-treated product containing the polyphenol and / or vitamin C of the present invention exhibits an effective improvement effect against various diseases.

  The acerola-treated product containing the polyphenol and / or vitamin C of the present invention has antioxidant activity, arteriosclerosis prevention activity, carcinogenesis inhibition activity, tumor growth inhibition activity, angiogenesis inhibition activity, antiallergic activity, antiinflammatory activity, cholesterol lowering activity, antibacterial activity, anti-caries activity, halitosis reduce activity, platelet aggregation inhibiting activity, anti-influenza virus activity, because they have gastric inhibitory activity and neutral fat absorption lowering activity, new formulations, foods utilizing these activities Useful as beverages, cosmetics, and the like.

  In addition, since acerola polyphenol has a vitamin C stabilizing activity, it is useful as a novel preparation, food, beverage, cosmetics and the like using this activity.

  Since the active ingredient of the preparation provided by the present invention is derived from acerola, which is a natural ingredient, there are few side effects.

It is the graph which showed the result of having measured the superoxide anion radical scavenging activity by ESR about the powder prepared in Example 3 and Example 4. FIG. It is the figure which showed the lipase inhibitory activity of the acerola polyphenol fraction. It is the figure which showed the time-dependent change of the plasma triglyceride value. Values are shown as mean ± standard error. It is the figure which showed the change of the blood total cholesterol of each group. Values are shown as mean ± standard deviation. It is the figure which showed the change of blood LDL-cholesterol of each group. Values are shown as mean ± standard deviation. It is the figure which showed the liver weight per 100g of body weight of each group. Values are shown as mean ± standard deviation. It is the figure which showed the total cholesterol amount per 1g of liver of each group. Values are shown as mean ± standard deviation.

  The present invention relates to a processed acerola containing polyphenol and / or vitamin C.

  The production area and varieties of acerola used in the present invention are not particularly limited, but examples of production areas include Okinawa and Brazil.

  The acerola processed product is acerola juice obtained by squeezing acerola fruit, acerola crushed product obtained by crushing and crushing acerola fruit, acerola extract obtained by extracting acerola fruit, or acerola fruit powdered or purified Although the acerola purified product etc. which are obtained by performing a process are pointed out, it is not necessarily limited to these. Moreover, a fruit refers to the whole fruit containing an edible part and a seed part.

  Acerola juice can be obtained by squeezing and collecting acerola fruit according to a conventional method.

  The acerola crushed material can be obtained by crushing and pulverizing the edible part and seed part of the acerola fruit or the edible part from which the seed part has been removed with a mixer or the like. In addition, the crushed material can be subjected to processing such as extraction and freeze-drying.

  The acerola extract can be obtained by extracting the acerola fruit with water, an organic solvent or the like. As the solvent, water is particularly preferable, and pure water, purified water, and the like can be used. When a hydrophilic organic solvent is used, for example, alcohol such as methyl alcohol, ethyl alcohol, glycerin, propylene glycol, 1,3-butylene glycol, acetone, tetrahydrofuran, acetonitrile, 1,4-dioxane, pyridine, dimethyl sulfoxide, N , N-dimethylformamide, acetic acid and other known organic solvents can be selected. It is particularly preferable to extract using methyl alcohol. The above hydrophilic organic solvents, particularly methyl alcohol and ethyl alcohol are preferably used as a mixture with water.

  The extraction conditions are not particularly limited, but the amount of the extraction solvent used is usually about 100 parts by weight to 1000 parts by weight, preferably about 200 parts by weight to 500 parts by weight with respect to 100 parts by weight of the fruit. The extraction temperature range is 0 ° C. to 120 ° C., preferably 20 ° C. to 50 ° C., from the viewpoint of efficient extraction. The extraction time is about 1 to 24 hours, preferably about 1 to 2 hours.

  An acerola purified product refers to an acerola juice, an acerola extract, or the like that has been subjected to a purification treatment as necessary. For example, an acerola extract can be obtained by appropriately concentrating to a desired concentration by removing the extraction residue by filtration or centrifugation after the extraction, or by further concentrating under reduced pressure. The acerola extract can also be prepared in a powder form by drying by freeze drying, spray drying or the like. At that time, it may be mixed with an excipient and dried. In addition, since the acerola extract contains a large amount of sugar and organic acid, it is also preferable to perform a purification step to remove them. Examples of the purification method include solvent fractionation, column fractionation, normal phase or reverse phase chromatography, ion exchange chromatography, gel filtration, and the like. A combination of these methods can also be used.

  The purified acerola product preferably contains acerola polyphenol at a relatively high concentration. Specific examples of the acerola polyphenol include anthocyanin pigments such as cyanidin-3-rhamnoside and pelargonidin-3-rhamnoside, quercitrin (quercetin-3-rhamnoside), isoquercitrin (quercetin-3-glucoside), hyperoside ( Quercetin glycosides such as quercetin-3-galactoside) and astilbine. These polyphenols may be provided as a mixture of a plurality of polyphenol compounds, or individual polyphenol compounds may be provided in an isolated form. A method for purifying the polyphenol compound at a high concentration is not particularly limited, and examples thereof include HPLC, synthetic adsorbent chromatography, ion exchange chromatography, gel filtration, and the like, and synthetic adsorbent chromatography is particularly preferable. In this case, as elution conditions, it is preferable to elute using, for example, a 10 to 50% ethanol solution. Furthermore, since the anthocyanin dye is stabilized under acidic conditions, it is particularly preferable to add hydrochloric acid or acetic acid to the eluate to make it acidic.

  An acerola polyphenol fraction obtained from acerola juice or an acerola extract is also preferably used in the present invention. Here, the acerola polyphenol fraction refers to a fraction containing a polyphenol compound eluted from the acerola juice or acerola extract by the above-mentioned various chromatographies under the above elution conditions. The acerola polyphenol fraction includes the eluate, its concentrate and its dried product.

  In addition, acerola polyphenol and vitamin C can be isolated and used according to a conventional method.

  The acerola-treated product containing polyphenol and / or vitamin C thus obtained has an antioxidant activity, an arteriosclerosis prevention activity, a carcinogenesis inhibition activity, a tumor growth inhibition activity, an angiogenesis inhibition activity, an antiallergic activity, an antiallergic activity, It has inflammatory activity, cholesterol-lowering activity, antibacterial activity, anti-cariogenic activity, bad breath odor reducing activity, platelet aggregation inhibitory activity, anti-influenza virus activity, gastric cancer inhibitory activity, and neutral fat absorption reducing activity.

  One of the preferred forms of the present invention is a form as a cholesterol lowering agent or a neutral fat absorption lowering agent. The acerola-treated product used in these forms preferably contains at least polyphenol. Cholesterol lowering agents and neutral fat absorption lowering agents are useful as preventive or therapeutic agents for hyperlipidemia. Neutral fat absorption reducing agents also have the effect of reducing body fat or maintaining it at a low level.

  There are two types of cholesterol in the blood: those synthesized in the liver and those taken from the diet. Those synthesized in the liver are contained in very low density lipoprotein (VLDL) and released into the blood. When it becomes (LDL), it is taken up through the receptor of a cell. Moreover, when the cholesterol content of hepatocytes increases, LDL uptake of cells is suppressed (receptor synthesis is suppressed), and as a result, blood cholesterol increases. From this, the effect of reducing the amount of cholesterol in the liver leads to the suppression of the blood cholesterol level. FIG. 7 shows that the processed acerola has the effect of reducing the amount of cholesterol in the liver.

  On the other hand, the neutral fat taken from the meal is digested and decomposed in the mouth and small intestine, and then passes through the small intestinal wall to become neutral fat again. They are then transported through the blood and spread throughout the body, including muscles and organs. At this time, the surplus that has not been used as energy is stored in adipose tissue or taken into the liver. Here, since body fat is a general term for adipose tissues attached to the body, suppression of neutral fat absorption leads to a decrease in body fat.

  The inventors have also found that acerola polyphenols are particularly superior as vitamin C stabilizers. Acerola polyphenol and vitamin C are considered to function as an antioxidant alone, but when they are combined, vitamin C is stabilized, and thus a synergistic antioxidant effect is exhibited. Therefore, it is preferable. Although the ratio of acerola polyphenol and vitamin C is not particularly limited, for example, the weight ratio is 1: 1000 to 10: 1, and more preferably 1:50 to 1: 1. In this embodiment of the present invention, the acerola polyphenol and vitamin C may be contained together in the acerola treated product, and are a mixture of acerola polyphenol and vitamin C obtained separately. There may be. Vitamin C is not limited to those derived from acerola, and natural or synthetic vitamin C (ascorbic acid) can be used.

  The acerola-treated product containing the polyphenol and / or vitamin C of the present invention is formulated in combination with a known pharmaceutical carrier, for example, an antioxidant, an arteriosclerosis preventive agent, a carcinogenesis inhibitor, a tumor growth inhibitor, an angiogenesis inhibitor May be administered as an agent, anti-allergic agent, anti-inflammatory agent, cholesterol-lowering agent, antibacterial agent, anti-cariogenic agent, bad breath odor reducing agent, platelet aggregation inhibitor, anti-influenza virus agent, stomach cancer inhibitor and neutral fat absorption reducing agent it can.

  Dosage forms are not particularly limited and may be appropriately selected as needed, typically tablets, capsules, granules, fine granules, powders, pills, solutions, syrups, suspensions, emulsions, elixirs Symptoms as oral preparations such as injections, or parenteral preparations such as injections, drops, suppositories, inhalants, transdermal absorbents, transmucosal absorbents, nasal preparations, enteral preparations, patches, ointments, etc. Depending on the, it is used alone or in combination. The addition amount of this preparation varies depending on various conditions such as the kind of the addition target and the form of use, but usually it is preferably used in the range of 0.01% by mass to 20% by mass with respect to the total addition target.

  The dosage of the preparation varies depending on the patient's age, body weight, degree of disease, administration route, but for oral administration, it is usually 10 mg to 3000 mg as a processed powder of acerola, and the number of administration is usually Oral administration is 1 to 3 times a day.

  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 disintegrants 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, when administering to such a formulation as a liquid agent, a syrup agent, a suspension agent, an emulsion, and an elixir agent, a flavoring agent and a coloring agent may be contained.

  Acerola polyphenols as vitamin C stabilizers and acerola polyphenols and vitamin C as antioxidants can also be formulated and administered as described above.

  The acerola-treated product containing the polyphenol and / or vitamin C of the present invention can also be added to foods in the form of solids, semi-solids, liquids and the like according to a conventional method. Although not necessarily limited, examples of the solid food include block confectionery such as biscuits and powdered food such as powder soup. In addition, dry powder of acerola extract can be used as food as it is. Examples of semi-solid foods include capsules and jellies. Examples of the beverage include fruit juice beverages, soft drinks, and alcoholic beverages. Moreover, it is good also as a form of the powdered drink diluted at the time of ingestion using liquid carriers, such as water. Moreover, polyphenols and / or acerola process containing vitamin C of the present invention is added to foods, it may be a so-called foods for specified health uses (e.g., diabetes, diabetic complications preventing food).

  In addition, for such foods, stabilizers, pH adjusters, sugars, sweeteners, various vitamins, minerals, antioxidants, excipients, solubilizers, binders according to conventional methods as necessary. , Lubricants, suspending agents, wetting agents, film-forming substances, flavoring agents, flavoring agents, coloring agents, fragrances, preservatives and the like can be added.

  Acerola polyphenols as vitamin C stabilizers and acerola polyphenols and vitamin C as antioxidants can also be used in the form of foods as described above.

  The acerola-treated product containing the polyphenol and / or vitamin C of the present invention can also be added to cosmetics such as skin lotions, cosmetic liquids, and emulsions, and makeup cosmetics such as foundations and lipsticks in accordance with conventional methods. The same applies to acerola polyphenol as a vitamin C stabilizer and acerola polyphenol and vitamin C as antioxidants.

  Acerola, which is a raw material for producing the preparation of the present invention, has been used for preparations, foods, beverages, cosmetics, and the like, and its safety has been established.

  Preferred examples of the present invention are shown below, but the scope of the present invention is not limited to these examples.

Example 1
100 g of acerola fruit from Brazil was extracted with 200 ml of 80% ethanol, centrifuged at 6000 rpm for 5 minutes and filtered. This operation was performed twice, and the supernatant was concentrated with an evaporator. It was possible to add excipients to the sample obtained here to make a powder, but in this state the sugar content was very high (40-60%), so yeast was used to supply the sugar content. And the polyphenol content was increased. Specifically, the sample concentrated with an evaporator was redissolved with distilled water so that the Brix value was 20 to 30%, 2% yeast (Saccharomyces cerevisiae) was added, and fermentation was performed at 32 ° C. for 20 hours. After fermentation, centrifugation and filtration were performed, and the supernatant was lyophilized to obtain 4.08 g of powder. As a result of component analysis of this powder, it was 2.4% glucose, 13.7% fructose, 35.2% vitamin C, and 1.72% total polyphenol.

Example 2
The acerola fruit from Brazil was squeezed to make 400 ml of fruit juice. This fruit juice was concentrated with an evaporator so that the Brix value was 20 to 30%, and 2% yeast (Saccharomyces cerevisiae) was added to the obtained concentrated fruit juice, followed by fermentation at 32 ° C. for 20 hours. After fermentation, centrifugation and filtration were performed, and the supernatant was lyophilized to obtain 15.9 g of powder. As a result of component analysis of this powder, glucose was 0%, fructose was 0%, vitamin C was 38.6%, and total polyphenol was 0.71%.

Example 3
100 g of acerola fruit from Brazil was squeezed, and the resulting residue was washed twice with 300 ml of distilled water. After washing, centrifugation was performed to recover 32 g of residue. 320 ml of distilled water was added to the resulting residue, finely pulverized, and then subjected to heat and pressure extraction at 121 ° C. for 1 hour. After extraction, the mixture was ice-cooled, centrifuged and filtered, and the supernatant was lyophilized to obtain 475 mg of powder. As a result of component analysis of this powder, glucose was 0%, fructose was 0%, vitamin C was 0.005%, and total polyphenol was 6.8%.

Example 4
The seeds were removed from 1654 g of Brazilian acerola fruit, the remaining edible portion was homogenized, and 3 times the amount of methanol was added and extracted for 1 hour. This operation was performed twice, followed by centrifugation, filtration, and freeze-drying to obtain 84.4 g. Since this sample contains about 40-60% sugar and about 10% vitamin C, column processing was performed to remove sugar and vitamin C in order to further increase the polyphenol content. Specifically, the obtained powder is dissolved again in distilled water, this solution is applied to a C18 cartridge column (Waters Sep-Pak Vac 35cc C18 cartridge column), washed with distilled water, and then washed with 0.2% TFA / methanol solution. Eluted. Thereafter, freeze drying was performed to obtain 2.79 g of powder. As a result of component analysis of this powder, glucose was 0%, fructose was 0%, vitamin C was 0%, and total polyphenol was 38.6%.

Example 5
100 g of acerola fruit from Brazil was squeezed to obtain fruit juice and residue. The residue was washed with distilled water, and this washing solution and fruit juice were combined and lyophilized. The obtained sample was dissolved again in distilled water, this solution was applied to a synthetic adsorption resin (Amberlite XAD7HP), washed with distilled water, and eluted with 0.2% TFA / methanol solution. Thereafter, freeze-drying was performed to obtain 120 mg of powder. As a result of component analysis of this powder, glucose was 0%, fructose was 0%, vitamin C was 0%, and total polyphenol was 40%.

Example 6
Next, the antioxidant activity of the acerola polyphenol-containing material was evaluated. Among the examples described above, the antioxidant activity of the powders prepared in Example 3 and Example 4 in which the activity of polyphenol itself was evaluated without the influence of vitamin C was evaluated.

[Evaluation of antioxidant activity]
Antioxidant activity was evaluated by the superoxide anion radical scavenging activity by the xanthine-xanthine oxidase reaction system, and ESR was used to detect the radical.

  Mix 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 add XOD After 45 seconds, ESR measurement was performed. In addition, the samples of Example 3 and Example 4 were prepared at the same concentration of 0.5 mg / ml.

The ESR apparatus used 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
Amplification rate: 100
Sweep time: 2 minutes, response time: 0.1 seconds Measurement temperature: room temperature

  The measurement results are shown in FIG. From FIG. 1, it was confirmed that the powders prepared in Example 3 and Example 4 all showed strong antioxidant activity.

Example 7
20 g of the powder prepared in Example 1, 60 g of lactose, 15 g of starch and 5 g of magnesium stearate were uniformly mixed to produce 1 tablet of 200 mg. This tablet contains 40 mg of acerola polyphenol-containing product per tablet. Tablets were similarly produced for the powders prepared in Examples 2 to 5.

  By using this preparation, an acerola polyphenol-containing substance of about 120 mg to about 480 mg per day can be administered to an adult.

Example 8
50 g of the powder prepared in Example 1, 40 g of lactose and 10 g of starch were uniformly mixed and dried and solidified, and then powders and granules were produced. This formulation contained 50% acerola polyphenol-containing material. For the powders prepared in Examples 2 to 5, powders and granules were similarly produced.

  By using this preparation, an acerola polyphenol-containing substance of about 150 mg to 450 mg per day can be administered to an adult.

Example 9
Preparation of acerola vitamin C and acerola polyphenol
A fruit pulverized product was prepared using a Waring blender while adding 5500 g of acerola fruit to 3500 g of purified water. Ethanol was added to the pulverized product so as to be 40% by weight, and the mixture was extracted with stirring overnight. The supernatant was collected by centrifugation, and then a clear extract was collected by filter filtration. Ethanol was removed from this extract by distillation under reduced pressure to obtain an acerola fruit extract concentrate.

  This concentrated solution was loaded onto an Amberlite XAD (product name: XAD7HP, Organo Corporation) column, which is a synthetic adsorbent, and acerola polyphenol was adsorbed onto the resin. At this time, the acerola fruit extract concentrated solution passed through the column was used as an acerola vitamin C solution. The acerola polyphenol adsorbed on the resin was eluted with ethanol. This eluate was dried by a vacuum distillation apparatus to obtain 10.3 g as an acerola polyphenol powder. The polyphenol content in the powder was measured by the foreign dennis method using catechin as a standard substance, and found to be 85.9%.

Example 10
Stability test of acerola vitamin C to which acerola polyphenol was added The acerola vitamin C solution prepared in Example 1 to a final concentration of 0.1% ascorbic acid was diluted with a citrate-phosphate buffer solution of pH 3.6. The acerola polyphenol powder prepared in Example 9 was dissolved so that the final polyphenol concentration would be 0.004%, 0.01% and 0.05%. All samples including samples not containing acerola polyphenol were filtered through a 0.2 μm filter, a storage test was performed at 40 ° C., and ascorbic acid concentrations at 0, 1, 4, 5, and 8 days were measured by the indophenol method. . The results are shown in Table 1 as the residual rate of ascorbic acid.

  From the results of Table 1, in the storage 1 day big difference even residual ratio in comparison with the additive-free acerola polyphenol addition group was observed, the higher the polyphenol concentration of additives in the storage 8 day ascorbic acid residual rate Therefore, it was confirmed that it is effective to add acerola polyphenol to an antioxidant containing acerola vitamin C.

Example 11
Stability test of ascorbic acid reagent to which acerola polyphenol was added Dilute an ascorbic acid (reagent special grade, manufactured by Wako Pure Chemical Industries, Ltd.) aqueous solution with a pH 3.6 citrate-phosphate buffer to a final concentration of 0.1% ascorbic acid. Here, the acerola polyphenol powder prepared in Example 9 was dissolved to a final polyphenol concentration of 0.004%, 0.01% and 0.05%. All samples including samples not containing acerola polyphenol were filtered through a 0.2 μm filter, a storage test was performed at 40 ° C., and ascorbic acid concentrations at 0, 1, 4, 5, and 8 days were measured by the indophenol method. . The results are shown in Table 2 as the ascorbic acid residual rate.

  From the results of Table 2, in the storage 1 day big difference even residual ratio is compared with the no additive acerola polyphenol addition group was observed, the higher the polyphenol concentration of additives in the storage 8 day ascorbic acid residual rate Therefore, it was confirmed that it is effective to add acerola polyphenol to an antioxidant containing acerola vitamin C.

Example 12
1) Preparation of acerola polyphenol fraction The seeds were removed from the acerola fruit, the remaining edible portion was homogenized, and 3 times the amount of methanol was added and extracted for 1 hour. This operation was performed twice, followed by centrifugation, filtration, lyophilization, and re-dissolution in distilled water. This solution was applied to a C18 cartridge column (BAKERBOND spe ^™ C18 Disposable column), washed with distilled water, eluted with 0.2% TFA / methanol solution, and concentrated to dryness to obtain an extract. The extract obtained here was used as an acerola polyphenol fraction (AP fraction). When the polyphenol content of the AP fraction was measured by the Folin-Denis method, the polyphenol content was 25%.

2) Lipase inhibitory activity of AP fraction
The lipase inhibitory activity of the AP fraction was measured. Swine pancreatic lipase (SIGMA) 50μl (0.5mg / ml), sample 50μl (1.0mg / ml, 0.5mg / ml), 1.25mM 4-methylumbelliferyl oleate (Fulka) 100μl are mixed and incubated at 37 ° C for 20 minutes Thereafter, 1 ml of 0.1N hydrochloric acid was added to stop the reaction. Furthermore, after adjusting the pH by adding 2 ml of 0.1 M trisodium citrate, fluorescence was measured at an excitation wavelength of 320 nm and a fluorescence wavelength of 450 nm.

  As apparent from FIG. 2, the AP fraction showed lipase inhibitory activity.

3) Animal test (single administration)
Using 7-week-old ICR male mice (CLEA Japan, Inc.), the triglyceride level in plasma after lipid loading was examined by dividing into two groups of 5 AP fraction administration groups and 5 control groups. Mice weighing approximately 30 g were fasted overnight and then cottonseed oil (2.5 ml / kg body weight) was administered. Next, the AP fraction (250 mg / kg body weight) dissolved in physiological saline was administered to the AP fraction administration group with an intragastric sonde, and physiological saline was administered to the control group. Blood was collected from the tail vein after 1, 2, 4 and 6 hours, and the triglyceride level in plasma was measured. Triglyceride E test Wako (Wako Pure Chemical Industries) was used for the measurement of plasma triglyceride levels. As a result, the AP fraction administration group tended to suppress the increase in triglyceride value compared to the control group (FIG. 3).

4) Animal test (continuous administration)
Lipid tolerance test was conducted using 7-week-old CD (SD) / IGS male rats (Nippon Charles River) divided into 3 groups: 6 AP administration groups, 6 control groups and 6 normal groups. It was. In the AP fraction administration group and the control group, cottonseed oil 0.5 ml / body was administered once daily for 14 consecutive days. To the AP fraction administration group, immediately after cottonseed oil administration, AP fraction (amount of 250 mg / kg body weight) dissolved in distilled water was administered by intragastric sonde. Distilled water was administered to the control group. In the normal group, cottonseed oil was not administered, but only distilled water was administered. Blood was collected from the orbital vein before loading with cottonseed oil on the 0th, 7th, and 14th days, and total cholesterol and LDL-cholesterol in plasma were measured. Each measurement result was evaluated based on the rate of change with the measurement result on day 0 as 100.

  After the test, the animals were fasted for 18 hours, laparotomized under Nembutal (40 mg / kg) anesthesia, and visually observed. Next, after removing the liver and measuring the weight, the total amount of cholesterol in the liver was measured.

  The total plasma cholesterol on the 7th and 14th days of the AP fraction administration group showed the same value as that of the normal group, and was significantly lower than that of the control group (FIG. 4). The same was true for LDL-cholesterol (FIG. 5). Macroscopic observation at the time of dissection revealed no abnormal sites in the main organs including the liver in any group. Regarding the liver weight, the normal group and the AP fraction administration group showed slightly lower values than the control group, but there was no significant difference (FIG. 6). The average value of total cholesterol in the liver showed the lowest value in the AP fraction administration group (FIG. 7).

Claims (2)

  A cholesterol-lowering agent containing acerola polyphenol as an active ingredient.   The cholesterol-lowering agent according to claim 1, wherein the acerola polyphenol is a polyphenol compound-containing fraction obtained from acerola juice or an acerola extract.

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