JP4545632B2 - Folic acid-containing composition and method for stabilizing folic acid - Google Patents

Description

  The present invention relates to stabilized folic acid-containing compositions, particularly liquid compositions useful as beverages, and methods for stabilizing folic acid.

1. What is folic acid;
Folic acid is a water-soluble vitamin belonging to the group B vitamins extracted from spinach and is abundant in vegetables such as carrots and pumpkins, fruits such as avocado, beans, milk, yellow eggs and liver.
Folic acid is important for red blood cell production, protein metabolism or nucleic acid (DNA and RNA) production, is essential for somatic cell differentiation and proliferation, and is also essential for sugar and amino acid consumption. It is already well known that it has many beneficial effects.
More specifically, for example, as one of the actions of folic acid, it is known that it functions to help the reaction in which homocysteine is converted into methionine necessary for protein synthesis in the cell.
Further, as described above, folic acid is indispensable for cell differentiation / proliferation and DNA synthesis, and is particularly important in the early stage of pregnancy when fetal cell proliferation is thriving. In recent years, many studies have revealed that taking sufficient folic acid before and after conception can reduce the risk of neural tube closure disorders such as spina bifida and anencephalopathy. 440 μg / day folic acid is recommended for pregnant women / day.

In addition, folic acid has a function of cooperating with vitamin B ₁₂ to help synthesize hemoglobin and produce red blood cells. Thus, a composition consisting of folic acid and vitamin B ₁₂ is useful as a hematopoietic agent. Furthermore, also since the required vitamin B ₁₂ in order to vitamin B ₆ essential for the production of red blood cells is properly absorbed, vitamin B ₁₂ for hematopoietic _{erythrocytes,} a combination of vitamins of B ₆ and folic important Have a meaning. Further, since zinc has an action of promoting absorption of folic acid, it is more effective to add zinc in addition to the folic acid composition.

With respect to folic acid, for patients with occlusive vascular disease or Alzheimer's disease at least moderately elevated blood homocysteine with at least moderately low blood folate and vitamin B ₁₂ concentrations of concentration, folic acid, folates or Derivatives, and optionally vitamin B ₁₂ , and optionally organic nitrates (eg isosorbide mononitrate or dinitrate), or ACE inhibitors or angiotensin II antagonists, or NEP / ACE inhibitors or mixtures of two or more thereof A method for the treatment of obstructive vascular disease or Alzheimer's disease comprising the administration of Accordingly, the pharmaceutical composition comprising folic acid and vitamin B ₁₂ are not only red blood cells increase multifunctional, such as described above is effective for the treatment of occlusive vascular disease or Alzheimer's disease. (For example, refer to Patent Document 1.)
Thus, folic acid is an important nutrient for the human body from fetuses to elderly people, and in recent years, the need for folic acid intake is increasing.

2. Folic acid compounds;
“Folic acid” refers to proteyl monoglutamic acid (synthetic folic acid) in a narrow sense, but in a broad sense, it is a coenzyme type, that is, a reduced type, a one-carbon unit substitution type, and a general term for these polyglutamic acids. Synthetic products (food additives) folic acid is said to have a higher utilization rate in vivo than food-derived folic acid. These synthetic products (food additives) can be obtained from BASF Takeda Vitamin Co., Ltd. and DSM Japan Co., Ltd. In the case of food origin, teas such as green tea, oolong tea, black tea, beans such as soybeans and broad beans, mustard greens, spinach, mizukake greens, nabara greens, green asparagus, spring chrysanthemum, sweet potatoes, broccoli, tinning What was extracted from fruits, such as vegetables, such as vegetables, strawberries, papayas, oranges, mandarin oranges according to a usual method, can be used. Extraction of folic acid from these plants is achieved by extracting with hot water or hot water in the case of tea leaves, and in the case of beans, vegetables and fruits, removing the insoluble fraction from the juice and collecting the extract. Is done.

3. Chemical structure of folic acid compounds;
The folic acid compound has the following pterin structure.

More specifically, folic acid is represented by the following formula.

  In the above, the asymmetric α-C atom of the glutamic acid residue may be present in the S-configuration (αS) or in the R-configuration (αR). The same is true for folic acid esters and other derivatives. These are referred to as (αS) -folate and (αR) -folate. Naturally produced folic acid corresponds to (αS) -folic acid.

Reduced tetrahydrofolic acid is represented by the following formula.

  In this case the asymmetric α-carbon atom of the glutamic acid residue is present in the S-configuration (αS) or in the R-configuration (αR) and the asymmetric carbon atom at position 6 in the tetrahydropterin residue May exist in the R-configuration (6R) or in the S-configuration (6S). The diastereomers of tetrahydrofolic acid can be referred to as (6S, αS)-, (6S, αR)-, (6R, αS)-and (6R, αR) -tetrahydrofolic acid. The same applies to tetrahydrofolate esters and other derivatives. These can be referred to as (6S, αS)-, (6S, αR)-, (6R, αS)-and (6R, αR) -tetrahydrofolate. Naturally occurring tetrahydrofolic acid corresponds to (6S, αS) -tetrahydrofolic acid. (For example, see Patent Document 2.)

In addition, 7,8-dihydrofolic acid is also known as one of the reduced folic acids.
Further, folic acid substituted at the 5-position and / or the 10-position is also known. For example, 5-methyltetrahydrofolic acid, 5,10-methylenetetrahydrofolic acid, 5-formyltetrahydrofolic acid, 5-formiminotetrahydrofolic acid, 10-formyltetrahydrofolic acid, 5,10-methenyltetrahydrofolic acid, and the like.
Some of the reduced folic acid can also be used as polymerized polyglutamyl folic acid (having 2 to 8 glutamic acid residues. For example, diglutamyl folic acid, triglutamyl folic acid, tetraglutamyl folic acid, pentaglutamyl folic acid and hexaglutamyl folic acid). Are known.
The chemical structural formula of polymerized polyglutamyl folic acid is as follows.

  Here, n is an integer of 1 to 7, and the nitrogen atoms at the 5th and 10th positions may be substituted with a methyl group, a formyl group or a formimino group, or the 5th and 10th positions are each a methylene group or a methenyl group. May be combined to form a ring.

4). Other vitamins B (vitamin B ₁₂ , vitamin B ₆ );
It has also been reported that the above folic acid does not sufficiently exert its effects without vitamins B ₆ , B ₁₂ and C.
In addition, these vitamin B ₁₂ and vitamin B ₆ are also known to have various beneficial effects.
For example, vitamin B ₁₂ is a water-soluble, exerts its effect by being combined with cobalt, erythrocyte formation or regeneration to prevent anemia, promote growth, fat, carbohydrates, the use of protein appropriate It has the action of controlling the sensitivity, suppressing the sensitive reaction to the stimulus, increasing the concentration and memory, and stabilizing the mind. Therefore, pernicious anemia and vitamin B ₁₂ is depleted, causing a neurological disorder. Vitamin B _{12 is} also unstable to light, acid, and alkali, like folic acid.
Vitamin B ₆ is a water-soluble vitamin, also known as pyridoxine, it is necessary to replenish daily from being excreted within 8 hours after ingestion. Vitamin B ₆ is properly absorbed in the presence of vitamin B ₁₂ and is essential for the production of antibodies and erythrocytes or gastric acid secretion, assists in the proper absorption of proteins and fats, and the essential amino acid tryptophan is vitamin B ₃ (niacin). ), Prevent various nerve and skin disorders, and promote normal synthesis of anti-aging nucleic acids. Accordingly, the vitamin B ₆ is depleted, anemia, seborrheic dermatitis, also causes glossitis. However, like other vitamin B, vitamin B ₆ also has problems such as being not always stable and difficult to store for a long time.

5). Stability of folic acid;
As described above, the need for folic acid intake has increased in recent years, but unfortunately folic acid is not always a stable compound. These folic acid compounds can exist stably in a limited region from weakly acidic to neutral regions. Therefore, under strong acidity, folic acid cannot be present stably. Moreover, since folic acid is decomposed by light, there is a problem in applying it to PET bottle beverages.
In this way, vitamins such as folic acid derived from natural products are very unstable, and are decomposed not only by acid but also by oxygen, heat, light, etc. over time, p-aminobenzoic acid, and other complex compounds It is known that the amount of folic acid tends to decrease with time. Therefore, in beverages such as commercially available can containers, paper containers, especially PET bottles, when drinking, folic acid is often contained in an amount considerably lower than the required intake.

6). Stabilization of folic acid;
As a method for solving these problems, for example, a method of blending erythritol and / or trehalose has been proposed as a method for stabilizing folic acid against acid. (For example, refer to Patent Document 3).
Further, as a method for stabilizing (6R)-, (6S)-or (6R, S) -5,10-methylenetetrahydrofolic acid and salts thereof, α-, β- or γ-cyclodextrin is added to these aqueous folic acid solutions. Or α-, β-
Alternatively, a method of stabilizing by adding a derivative of γ-cyclodextrin has been proposed. (For example, see Patent Document 4)
In addition, as a method for stabilizing 5,6,7,8-tetrahydrofolic acid and its derivatives, these compound powders should be stored at low temperature in the presence of an inert gas in the presence of an oxygen scavenger, blocking the outside air. A stabilization method using the above has also been proposed. (For example, refer to Patent Document 5.)
However, although these stabilization methods can achieve the purpose in terms of stabilization of folic acid, compounding agents such as cyclodextrin did not play a role more than the third component for stabilization.
JP-T-2001-504104 Special table 2003-504370 gazette Japanese Patent Laid-Open No. 2002-097141 Japanese Patent Application Laid-Open No. 07-018084 Japanese Patent Application Laid-Open No. 06-065243

The object of the present invention is not only to stabilize folic acid, but also to stabilize these folic acids by using a pharmaceutically effective third component, and the other object is to provide a pharmaceutically effective third component. It is to provide a stabilized vitamin B-containing composition, particularly a folic acid-containing composition.

As a result of intensive studies to solve the above problems, the present inventors have completed the present invention. More specifically, the present inventors have examined food materials that have been used for food for many years and have been confirmed to be safe for the human body, and those that have utility value as stabilizing components for vitamin Bs, particularly folic acid. investigated. As a result, the present inventors have found that the polyphenol compound has a strong stabilizing action against vitamin Bs, particularly folic acid, particularly a stabilizing action against light such as folic acid in an aqueous solution.
More specifically, the present invention is as follows.

1. 1 mol of folic acid, folic acid containing at least 0.05 mol of tea-derived epigallocatechin gallate, epicatechin gallate, epigallocatechin and epicatechin A method of stabilizing the composition.
2. The method for stabilizing a folic acid-containing composition according to claim 1, wherein the folic acid-containing composition is a beverage composition.
3. The method for stabilizing a folic acid-containing composition according to claim 2, wherein the folic acid concentration is 24 μg / 500 mL to 4,800 μg / 500 mL, and the catechin content is 100 mg / 500 mL to 1,000 mg / 500 mL.
4). The method for stabilizing a folic acid-containing composition according to any one of claims 1 to 4, wherein the catechin is epigallocatechin gallate.
5). The folic acid is at least one folic acid selected from folic acid, dihydrofolic acid, tetrahydrofolic acid and derivatives thereof, polymerized polyglutamate and derivatives thereof, ester forms thereof and salts thereof. A method for stabilizing a folic acid-containing composition as described.

  In the present invention, folic acid can be stabilized by using a polyphenol compound as a stabilizer, and the efficacy / effect and drug efficacy of the polyphenol itself can be expected. In addition, since polyphenols are contained in green tea and the like and have been confirmed to be safe, there is no problem even when ingested for a long time.

Hereinafter, embodiments of the present invention will be described.
The meanings of terms used in the present specification are as follows.

“Stabilized folic acid-containing composition” means a folic acid-containing composition in which folic acid can exist in a stable state, and morphologically contains folic acid and polyphenols for addition to beverages and foods. A solid composition (powder, tablet, etc.) as a raw material for blending, a liquid composition such as a beverage obtained by adding these solid powder compositions to a beverage stock solution, or a solid powder composition such as gum or candy It may be present as a solid composition such as gum or candy in addition to food. From the viewpoint of use, the composition is a pharmaceutical composition; a quasi-drug composition; a so-called health food (supplement) having a pharmacological effect; a composition for blending to be added to a beverage or food (food addition) A beverage composition obtained by adding the blending composition; or a food composition obtained by adding the blending composition to a food (nutrient functional food, food for specified health use, functional food). And so on.

When a pharmaceutical composition, a quasi-drug composition, or a so-called health food (supplement) having a pharmacological effect is added, a formulation such as a tablet, capsule, granule or the like is added according to a conventional method by appropriately adding a compounding agent such as a binder. By making it, it can make it easier to ingest.
When used as a composition for blending, it may be a powdery raw material, or may be processed into a tablet, capsule, granule or the like according to a conventional method.

The beverage composition is preferably a containerized beverage composition, for example, tea beverages such as green tea, oolong tea, and tea; juices such as apple juice, grapefruit juice, and orange juice; fruit juice blended beverages; vegetable juice blended beverages; coffee Beverages; Cocoa drinks; Milk drinks such as milk; Carbonated drinks such as cola and cider; Sports drinks; Soy milk; Mineral water; Near water drinks; Diet support drinks; Moreover, alcoholic beverages, such as beer, a sparkling wine, wine, and sake, may be sufficient. These beverage compositions can be produced by blending an appropriate amount of the above composition for blending into a tea beverage such as green tea, oolong tea, black tea or the like (for example, tea extract). In the case of a tea beverage, since the tea beverage itself contains polyphenols such as catechin, it can be produced by blending an appropriate amount of folic acid within the range that can be stabilized as a result.

The food composition is a confectionery, bread, yogurt, jelly, frozen confectionery, candy, gum or the like obtained by blending an appropriate amount of the above composition for blending into raw materials and processing it according to a conventional method.

The “stabilized folic acid-containing composition” is preferably a beverage composition such as juices; fruit juice-containing beverages; vegetable juice-containing beverages; milk beverages; tea beverages.

In addition, the above-mentioned “beverage composition” includes various sweeteners, acidulants, inorganic acid salts, inorganic acid salts, organic acids, nutrient fortifiers, oxidizing agents that can be used as food additives in addition to folic acid and polyphenol compounds. Inhibitors, fragrances, pigments, emulsifiers, preservatives, seasonings, lipids, proteins, peptides, polysaccharides, dietary fibers, fruit juice extracts, vegetable extracts, nectar extracts, pH adjusters, etc. alone Or you may mix | blend together.
Examples of the sweetener include sugar, glucose, fructose, isomerized liquid sugar, glycyrrhizin, stevia, aspartame, fructooligosaccharide, cyclic oligosaccharide, artificial sweetener and the like.
Examples of the sour agent include citric acid, tartaric acid, malic acid, lactic acid, fumaric acid, and phosphoric acid in addition to fruit juices extracted from natural ingredients.
Examples of inorganic acids and inorganic acid salts include phosphoric acid, disodium phosphate, sodium metaphosphate, and sodium polyphosphate. Examples of organic acids and organic acid salts include citric acid, succinic acid, itaconic acid, malic acid, sodium citrate and the like.
In addition, β-carotene, vitamin A, vitamin B other than folic acid (for example, vitamin B ₁ , vitamin B ₂ , niacin, pantothenic acid, etc.), biotin, vitamin C, vitamin D, vitamin E , Coenzyme Q10, α-lipoic acid, carnitine, calcium, iron, magnesium, zinc, copper and the like.
Containers used for container-packed beverages, like ordinary beverages, are molded containers mainly composed of polyethylene terephthalate (so-called PET bottles), metal tubes, paper containers combined with metal foil and plastic films, bottles, etc. It can be provided in the usual form. A PET bottle is preferable. Moreover, the container-packed drink here means what can be drunk without dilution.

“Folic acid” refers to folic acid described above; tetrahydrofolic acid, dihydrofolic acid; substituted tetrahydrofolic acid substituted at the 5-position and / or 10-position; and polymerized polyglutamate (having 2 to 8 glutamic acid residues). means. Specific examples include folic acid, 7,8-dihydrofolic acid, 5,6,7,8-tetrahydrofolic acid, 5-methyltetrahydrofolic acid, 5,10-methylenetetrahydrofolic acid, 5,10-methenyltetrahydrofolic acid, 5, 10-formiminotetrahydrofolic acid, 5-formyltetrahydrofolic acid (leucovorin), 5-formyliminotetrahydrofolic acid, 10-formyltetrahydrofolic acid, polymerized polyglutamylfolic acid (having 2 to 8 glutamic acid residues), esters thereof The body and their salts and mixtures thereof.
Examples of the polymerized polyglutamyl folic acid include dihydrotayl, triglutamyl, tetraglutamyl, pentaglutamyl and hexaglutamyl derivatives of tetrahydrofolic acid such as 5-formimino- (6S) -tetrahydrofolic acid.
Also, unless otherwise limited, the term “folic acid” encompasses both enantiomers (αS) and (αR), and for tetrahydrofolic acid, its diastereomers ((6S, αS), (6S, αR) , (6R, αS) and (6R, αR)).
The tetrahydrofolic acid derivative and the derivative in the polymerized polyglutamyl folic acid derivative mean a substituted product at the 5-position and / or the 10-position.
In the present invention, preferred folic acid is folic acid classified into the above-mentioned synthetic product (food additive) from the viewpoint of in vivo utilization efficiency and content control, specifically, BASF Takeda Vitamin Co., Ltd. Available from DSM Japan Inc. From the viewpoint of chemical structure, pteroyl monoglutamic acid is particularly preferred. In addition, if derived from natural products, teas such as green tea, oolong tea, black tea, beans such as soybeans, broad beans, mustard vegetables, spinach, mizukake vegetables, nabara vegetables, green asparagus, spring chrysanthemum, sweet potatoes, broccoli, What can be obtained (extracted) from fruits such as vegetables such as chingen vegetables, strawberries, papayas, oranges and mandarin oranges can be used. Methods for extracting folic acid from these foods are known, such as a method of extracting with hot water or hot water in the case of tea leaves, a method of recovering the extract by removing the insoluble fraction from the juice in the case of beans, vegetables and fruits. The general method can be adopted. In some cases, it is possible to use the juice as it is without extraction.

“Polyphenols” mean a series of compounds having a phenol structure having a plurality of hydroxyl groups, and examples include flavones, flavonols, flavanones, flavanols, anthocyanidins, and isoflavones. Specifically, as the flavone, rutin, luteolin, etc., as flavonol, quercetin, kaempferol, etc., as flavanone, hesperidin, naringin, etc., as flavanol, catechins, etc., as anthocyanidin, cyanidin, etc., Examples of isoflavones include genistein, genistin, daidzein, and examples of catechins include epicatechin, epigallocatechin, epicatechin gallate, epigallocatechin gallate, catechin, gallocatechin, catechin gallate, and gallocatechin gallate. it can.
In addition, chlorogenic acids, protoanthocyanidins, theaflavins, thealvidins and the like can be mentioned.
The preferred polyphenol compounds are catechins, particularly preferably tea-derived epigallocatechin gallate, epicatechin gallate, epigallocatechin or epicatechin, and more preferably epicatechin gallate.
The origin of the above polyphenols is not particularly limited, but among them, tea, coffee, cocoa, wine, grapes, apples, berries, cherry, lemon, orange, sikhshire, grapefruit, melon, onion, broccoli, From spinach, carrots, celery, eggplant, garlic, cabbage, me cabbage, cucumber, buckwheat, brown rice, soybeans, red beans, barley, basil, tarragon, mint, oregano, thyme, sage, rosemary and mixtures thereof, especially Those derived from tea are suitable because they can be easily applied as beverages. In particular, catechins contained in tea are more preferable because of their excellent water solubility and their track record as favorite beverages that are drunk on a daily basis.
As a method for extracting polyphenols from these plant raw materials, a known or well-known general method such as a method of extracting with water or hot water and filtering the residue or a method of removing insoluble substances from the juice can be adopted. .

“Vitamin B ₁₂ ” and “Vitamin B ₆ ” are not particularly limited, and may be commercially available products produced industrially or “vitamin B ₁₂ ” or “vitamin B” contained in food. ₆ "may be a per se.

  The “zinc” is not particularly limited as long as it is zinc that can be used as a food or as a pharmaceutical, particularly zinc having absorbability in the body.

Next, the blending amounts of folic acid and polyphenols will be described.
According to the dietary intake standard (2005 edition) published by the Ministry of Health, Labor and Welfare, the recommended daily intake of folic acid depends on body weight, age, health, sex, and other factors, but 240 μg / day for adults and pregnant women 440 μg / day.
Considering the amount of folic acid taken from daily foods and drinks, the minimum content of folic acid contained in the composition of the present invention is about 1/10 of the recommended amount, that is, 24 μg / day. It is estimated that folic acid is excreted from the body and is harmless, and its utilization rate in vivo is estimated to be 50% (folic acid is decomposed by digestive enzymes in the small intestine and absorbed into the body. In view of the fact that the utilization rate in vivo is estimated to be 50% through various stages, the maximum content can be about 20 times the recommended adult dose, that is, 4,800 μg.
From this viewpoint, the appropriate folic acid intake from the folic acid-containing composition of the present invention is 24 μg / day to 4,800 μg / day, preferably 100 μg / day to 4,800 μg / day, particularly preferably 200 μg / day to A guideline of 4,800 μg / day can be used.
Specifically, in the case of a beverage composition, one or two drinks per day are taken as 24 μg / 500 mL to 4,800 μg / 500 mL, preferably 100 μg / 500 mL to 4,800 μg / 500 mL, particularly preferably Is 200 μg / 500 mL to 4,800 μg / 500 mL. In the case of a PET bottle beverage, since there is a high possibility of exposure to sunlight, an even higher concentration of 300 μg / 500 mL to 4,800 μg / 500 mL is preferable.
In the case of pharmaceutical compositions, supplements, and food compositions, the content may be appropriately determined based on the recommended amount per day.
In the invention of the folic acid-containing composition of the present invention, the content of polyphenols relative to folic acid is important from the viewpoint of stabilization, but the content of folic acid itself is simply based on the above recommended amount. It can be determined as appropriate.

  “Stabilization” in “polyphenols in an amount sufficient to stabilize folic acid” means at least stability to folic acid when placed under a fluorescent lamp (artificial meteor) at 5,000 lux at 20 ° C. It means that the folic acid can be found, preferably when the folic acid is present for at least 1 week under the above conditions, and the presence of folic acid is at least 50%, particularly preferably 70% or more.

The lower limit of the blend of polyphenols to be blended in the composition of the present invention means the minimum amount necessary for stabilizing folic acid, and the minimum blending amount (coexistence amount) is the same as that of folic acid as will be apparent from the examples described later. The molar ratio is 0.05 times (in the case of ECG, 0.05 times in weight ratio), preferably 0.1 times in molar ratio to folic acid (in the case of ECG, 0.1 times weight ratio). On the other hand, polyphenols such as catechins are non-toxic and effective in reducing body fat, but if added excessively unnecessarily, problems such as bitterness and astringency peculiar to catechins occur. The content of is preferably 100 mg / day to 1,000 mg / day (in beverages, 100 mg / 500 mL to 1,000 mg / 500 mL).
For example, when folic acid is 100 μg / day to 4,800 μg / day (100 μg / 500 mL to 4,800 μg / 500 mL), the minimum amount of polyphenols required for stabilization is 0.05 times ECG, 5 μg / day. To 240 μg / day (in the case of beverages, 5 μg / 500 mL to 240 μg / 500 mL), preferably 0.1 μg / day to 480 μg / day (10 μg / 500 mL to 480 μg / 500 mL).
However, from the viewpoint of sufficiently ingesting the necessary polyphenols, the polyphenol content is preferably 100 mg / day to 1,000 mg / day (100 mg / 500 mL to 1000 mg / 500 mL for beverages). With such a content, as is apparent from the above, the intake of polyphenol is sufficient, and as a result, folic acid is naturally stabilized.

Similarly, according to the Ministry of Health, Labor and Welfare's dietary intake standard (2005 version), the recommended daily amount of “Vitamin B ₁₂ ” is 2.4 μg / day, while the recommended amount of folic acid is 240 μg (440 μg for pregnant women). For this reason, when blending “vitamin B ₁₂ ”, it is preferable to blend 0.005 to 0.05 times by weight the amount of folic acid. Preferably, the standard can be 1.2 μg / day to 12 μg / day. Specifically, in the case of a beverage composition, it is 1.2 μg / 500 mL to 12 μg / 500 mL, assuming that one or two drinks per day.

Similarly, according to the Ministry of Health, Labor and Welfare's dietary intake standard (2005 edition), the recommended daily dose of “vitamin B ₆ ” is 1.4 mg for adult males, while the recommended intake of folic acid is 240 μg (440 μg for pregnant women). Therefore, when blending “vitamin B ₆ ”, it is preferable to blend 3 to 10 times the amount of folic acid.

The recommended daily amount of “zinc” is 9 mg for adult males and 7 mg for females. On the other hand, the recommended amount of folic acid is 240 μg (440 μg for pregnant women), and zinc is not essential in the composition of the present invention. Since it plays an auxiliary role, when blending “zinc”, it is preferable to blend 10 to 50 times by weight thereof with respect to folic acid.

  The folic acid-containing composition of the present invention, particularly the beverage composition, can further enhance the folic acid stabilization effect by combining storage under light-shielding conditions. Storage under light-shielding conditions means that the beverage itself is not directly exposed to light, such as storage of the beverage in a transparent or opaque container in a light-shielded box or storage of the beverage in a light-shielding container. Such storage. Here, the light-shielding condition refers to a state in which about 50% or more, preferably about 70% or more, more preferably about 90% or more of visible light is cut. In particular, since light with a short wavelength has high energy, it is desirable that light having a wavelength of about 340 nm or less is cut by about 80% or more, more preferably about 90% or more. For example, when filled in a metal can container, the above light shielding conditions are substantially satisfied.

Folic acid used in the present invention can be quantified by high performance liquid chromatography, microbial assay, or ELISA. Further, the polyphenols used in the present invention can be quantified by a high performance liquid chromatography method, an iron tartrate colorimetric method, a foreign dennis method, a foreign thiocult method, or the like.

  EXAMPLES Hereinafter, although this invention is demonstrated further in detail based on an Example, this invention is not limited to these Examples.

1. Folic acid stability test over time;
In 20 mL of 50 mM phosphate buffer (pH 6), folic acid (manufactured by Kanto Chemical Co., Inc., molecular weight 441) was dissolved to a concentration of 100 μM (final concentration). In this folic acid solution, epigallocatechin (EGC; molecular weight 306) (manufactured by Funakoshi Co., Ltd.) (sample 1), epigallocatechin gallate (EGCG; molecular weight 458) (manufactured by Funakoshi Co., Ltd.) (sample 2) were used as polyphenol compounds. ), Epicatechin (EC; molecular weight 290) (manufactured by Funakoshi Co., Ltd.) (sample 3) and epicatechin gallate (ECG; molecular weight 442) (manufactured by Funakoshi Co., Ltd.) (sample 4) were added at 10 μM (final concentration), Samples 1 to 4 were used. The folic acid was tested for stability over time by placing these samples under an artificial weather apparatus (manufactured by Nippon Medical Chemical Co., Ltd.) at 25 ° C. and 5,000 lux.
In addition, the weight conversion value of each component here is folic acid; 44.1 mg, EGC; 3.06 mg, EGCG; 4.58 mg, EC 2.9 mg, ECG; 4.42 mg.
At the same time, as a comparative example, a folic acid solution placed in the dark (no light irradiation) (sample 5), a folic acid solution without a polyphenol compound placed under an artificial meteor at 25 ° C. and 5,000 lux (sample 6) The folic acid solution (sample 7) to which only 100 μM (final concentration) of vitamin C (L-ascorbic acid; manufactured by Wako Pure Chemical Industries, Ltd.) was added was also tested.
After placing the sample for a certain period of time, folic acid was measured over time. As a control, the same concentration of vitamin C was used for comparison.

2. Measurement of folic acid;
The measurement of folic acid was performed using high performance liquid chromatography equipped with an ultraviolet absorption detector. Using an ODS column (made by Wako Pure Chemical Industries, Ltd.) having an inner diameter of 4.6 mm and a length of 150 mm and a particle size of 3 μm, 12% acetonitrile (including 0.05% phosphoric acid) was passed at a flow rate of 0.8 mL / min. . The sample was passed through a 0.45 μm filter, and folic acid was quantified by monitoring ultraviolet absorption at 290 nm.

3. Catechin measurement;
Catechin was measured using high performance liquid chromatography equipped with an electrochemical detector. Using an ODS column (made by Wako Pure Chemical Industries, Ltd.) having an inner diameter of 4.6 mm and a length of 150 mm and a particle size of 3 μm, 12% acetonitrile (including 0.05% phosphoric acid and 0.01% ethylenediaminetetraacetate disodium) was used. The liquid was passed at a flow rate of 0.8 mL / min. The sample was passed through a 0.45 μm filter, and each catechin was quantified by monitoring the reaction at an applied voltage of 500 mV.

4). Results of folic acid stability over time;
The results (residual amount of folic acid) in the time stability test are shown in FIG.
All of the folic acid in the folic acid solution without the polyphenol compound (Sample 6) disappeared on the first day under light irradiation conditions. On the other hand, in the folic acid solution to which only vitamin C was added (sample 7), the deterioration of folic acid was somewhat suppressed by the addition of vitamin C, but various catechins were added as polyphenol compounds when compared at the same concentration (10 μM). In samples 1 to 4, folic acid remained longer. Therefore, it can be said that the stability of folic acid was increased by the addition of polyphenols such as catechin.
In addition, as a result of examining the difference in stabilization of folic acid depending on the type of catechin contained in the leaves, all catechins are 0.1 mol times the concentration of folic acid (in the case of ECG, 0.01 times by weight). It had the effect of improving stability. Moreover, it turned out that the effect of stabilization has a high effect which stabilizes folic acid in order of ECG>EC>EGCG> EGC.

1. Concentration dependency test of polyphenol The concentration dependency effect when the concentration of epicatechin gallate (ECG) was changed as a polyphenol compound was examined. The results are shown in FIG.
In 20 mL of 50 mM phosphate buffer (pH 6), folic acid (manufactured by Wako Pure Chemical Industries, Ltd.) 100 μM (final concentration: 44.1 mg / L) was dissolved. In this folic acid solution, epicatechin gallate (ECG; molecular weight 442) (manufactured by Funakoshi Co., Ltd.) 5 μM (sample 8.2.21 mg), 10 μM (sample 9.4.42 mg), 20 μM (sample 10) was used as a polyphenol compound. 8.84 mg) (final concentration) was added, and the samples were placed under a 5,000-lux artificial meteor (manufactured by Nippon Kayaku Machinery Co., Ltd.) to test the folic acid stability over time.
At the same time, as a comparative example, a folic acid solution placed in the dark (no light irradiation) (sample 11) was also tested in the same manner.
After placing the sample for a certain period of time, folic acid was measured over time.
As a result, the stabilizing effect of folic acid was observed even in the 5 μM test group with the lowest concentration (molar ratio 0.05 times, in the case of ECG, 0.05 times weight ratio). The effect depends on the amount of addition, and is 10 μM (0.1 times in terms of molar ratio, about 0.1 times in terms of weight in the case of ECG) and 20 μM. A higher folic acid stabilization effect was found at twice the amount of 20 μM.

A beverage containing folic acid was prepared and tested for stabilization with polyphenols. The manufacturing method of each sample is as follows.

Example A
Green tea extract (Teafuran 90S, manufactured by ITO EN Co., Ltd.) was dissolved in ion-exchanged water so as to be 1 g / L, and folic acid (manufactured by Kanto Chemical Co., Ltd.) was added to a final concentration of 22 mg / L. 10 mL of this solution was placed in a threaded test tube and autoclaved at 121 ° C. for 20 minutes.

Example B
1 L of hot water was added to 100 g of oolong tea leaves, and the tea leaves were removed after standing for 10 minutes. Thereafter, the mixture was centrifuged at 5000 × g for 20 minutes at room temperature, and concentrated and dried. This oolong tea extract powder was dissolved in ion-exchanged water so as to be 4 g / L, and folic acid and vitamin C were added to final concentrations of 4.4 mg / L and 0.3 g / L, respectively. 10 mL of this solution was placed in a threaded test tube and autoclaved at 121 ° C. for 20 minutes.

Example C
1.25 L of 95 ° C. hot water was added to 100 g of tartary buckwheat and extracted for 20 minutes. The extract was cooled, then filtered with buckwheat and meshed, and centrifuged at 3000 rpm for 15 minutes. The buckwheat extract was concentrated and lyophilized. This buckwheat extract powder was suspended in ion-exchanged water so as to be 1 g / L, and folic acid was added so as to be 4.4 mg / L. 10 mL of this solution was placed in a threaded test tube and autoclaved at 121 ° C. for 20 minutes.

Example D
1 L of ion exchange water was added to 10 g of barley tea and boiled for 10 minutes. To this barley tea extract, folic acid was added to a concentration of 33 mg / L, and 10 mL of this solution was placed in a test tube with a screw and autoclaved at 121 ° C. for 20 minutes.

Comparative Example Folic acid was added to ion-exchanged water to a concentration of 44 mg / L, and autoclaved at 121 ° C. for 20 minutes. The stability of folic acid over time was tested by placing the prepared sample in an climatic instrument at 25 ° C. and 5,000 lux for 8 days.

Measurement of polyphenols Polyphenols were measured using the foreign dennis method and the high performance liquid chromatography method. The foreign dennis method used tannic acid (Nacalai Tesque Co., Ltd.) as a standard substance.

  The results are shown in Table 1.

(result)
In the comparative example, the residual ratio of folic acid was 0, while the folic acid-containing beverages of Examples A, B, C, and D were about 70%, 70%, 85%, and 55%, respectively. , B, and C maintained high folic acid residual ratios of about 70% and 85%.

  According to the present invention, folic acid can be stably retained by the coexistence of polyphenols, particularly catechins. Therefore, the present invention can be effectively applied to PET bottle-packed folic acid-containing beverages that are exposed to irradiation conditions in stores or the like.

It is a graph which shows the folic acid residual rate at the time of adding various catechins as a polyphenol compound. Example 1 It is a graph which shows the relationship between epicatechin gallate density | concentration and the folic acid residual rate at the time of adding epicatechin gallate as a polyphenol compound. (Example 2)

Claims (5)

1 mol of folic acid, folic acid containing at least 0.05 mol of tea-derived epigallocatechin gallate, epicatechin gallate, epigallocatechin and epicatechin A method of stabilizing the composition. The method for stabilizing a folic acid-containing composition according to claim 1, wherein the folic acid-containing composition is a beverage composition. The method for stabilizing a folic acid-containing composition according to claim 2, wherein the folic acid concentration is 24 µg / 500 mL to 4,800 µg / 500 mL, and the catechin content is 100 mg / 500 mL to 1,000 mg / 500 mL. The method for stabilizing a folic acid-containing composition according to any one of claims 1 to 4, wherein the catechin is epigallocatechin gallate. The folic acid is one or more types of folic acid selected from folic acid, dihydrofolic acid, tetrahydrofolic acid and derivatives thereof, polymerized polyglutamate and derivatives thereof, esters thereof and salts thereof. A method for stabilizing a folic acid-containing composition as described.