JP7201142B2 - Folic acid-containing acidic composition with excellent stability of folic acid - Google Patents

Description

The present invention relates to folic acid-containing acidic compositions with reduced folic acid degradation rate and/or improved folic acid stability.

Folic acid is one of the essential vitamins for living organisms, and folic acid functions as a coenzyme in vivo and participates in protein biosynthesis and the like. Thus, folic acid deficiency can lead to various diseases and disorders (eg, megaloblastic anemia, neuropathy, intestinal dysfunction, etc.). Furthermore, it has been reported that fetal neural tube abnormalities that can be caused by maternal nutritional deficiency during pregnancy can be prevented by administering folic acid, and that folic acid exhibits a protective effect against cancer, especially epithelial cancer. (Non-Patent Documents 1 and 2). Therefore, in recent years, the importance of folic acid has attracted attention.

According to the "Dietary Reference Intakes for Japanese (2015 edition)" (Non-Patent Document 3) published by the Ministry of Health, Labor and Welfare, the recommended intake of folic acid is 240 μg/day for adults, 480 μg/day for pregnant women, and 340 μg/day for lactating women. It is said to be a day. Although folic acid is contained in various foods, nearly half of the folic acid in foods is lost by cooking or heating. Absent.

Therefore, supplements and foods and drinks containing high-concentration folic acid have been developed and sold, and are gaining popularity because they can ingest the above-mentioned recommended intake of folic acid more efficiently than ingesting food. In particular, liquid supplements and beverages containing high-concentration folic acid are highly popular because of their ease of ingestion and their appeal as luxury goods.

On the other hand, folic acid is stable in a dry state, but it is known that its stability decreases in an aqueous solution, particularly in an acidic region suitable for beverages (Patent Documents 1 and 2). Patent Document 1 discloses a technique for improving the stability of folic acid in an acidic aqueous solution by forming a complex of folic acid with lactoferrin. In Patent Document 2, by blending an inorganic salt or an organic salt of one or more metals selected from the group consisting of calcium, copper, and zinc into a folic acid-containing beverage containing benzoic acids as preservatives, the beverage is obtained. Techniques have been disclosed to improve the stability of folic acid in

However, these methods are based on the premise that lactoferrin and benzoic acids are blended together with folic acid in the composition, and the use and form of supplements and beverages may be limited. There is a strong need for new means that can improve the stability of folic acid in acidic aqueous solutions.

Japanese Patent No. 4339979 Japanese Unexamined Patent Application Publication No. 2011-55828

Czeizel, A.; E. , J. Pediat. Gastroenterol. Nutr. , vol. 20, pp. 4-16, 1995 Glynn. S. A. , D. Albanes, Nutr. Cancer, vol. 22, p. 101, 1994 Dietary Reference Intakes for Japanese (2015 edition), Ministry of Health, Labor and Welfare, March 2014

An object of the present invention is to provide a folic acid-containing acidic composition in which the rate of decomposition of folic acid is reduced and/or the stability of folic acid is excellent.

As a result of intensive studies by the present inventors to solve the above problems, the presence of a protic organic solvent increases the rate of decomposition of folic acid in an acidic composition, and/or found to be less stable. It has also been found that reducing the amount of protic organic solvent present in the acidic composition reduces the rate of folic acid degradation and/or improves its stability.

The present invention is based on these findings and includes the following inventions.
[1] An acidic composition containing folic acid, wherein the content of the protic organic solvent is less than 0.1% by weight.
[2] The composition of [1], wherein the protic organic solvent is ethanol, glycerin, methanol, acetic acid, formic acid, butylamine.
[3] The composition of [1] or [2], which is a packaged beverage or a packaged jelly beverage.
[4] A method for producing an acidic composition containing folic acid, comprising blending raw materials such that the blending amount of the protic organic solvent is less than 0.1% by weight.
[5] A method for improving the stability of folic acid in an acidic composition, comprising reducing the amount of protic organic solvent contained in the composition to less than 0.1% by weight.
[6] The method of [4] or [5], wherein the protic organic solvent is ethanol, glycerin, methanol, acetic acid, formic acid, or butylamine.
[7] The method of any one of [4] to [6], wherein the acidic composition is a packaged beverage or a packaged jelly beverage.

According to the present invention, it is possible to provide a folic acid-containing acidic composition with a reduced folic acid decomposition rate and/or excellent folic acid stability. According to the present invention, it is possible to provide a folic acid-containing acidic composition that is excellent in storage stability and that enables efficient ingestion of folic acid in the recommended intake amount.

The present invention relates to an acidic composition containing folic acid and having a reduced content of protic organic solvents.

あるいはその誘導体又は医薬品や飲食品において許容可能なその塩を意味する。誘導体としては例えば、複数のグルタミン酸が結合したポリグルタミン酸型等が挙げられるが、これらに限定はされない。 In the present invention, "folic acid" is one of the water-soluble vitamins of the vitamin B complex, a compound represented by the following structural formula called pteroylmonoglutamic acid,

Alternatively, it means a derivative thereof or a salt thereof acceptable in pharmaceuticals, food and drink. Examples of derivatives include, but are not limited to, polyglutamic acid forms in which multiple glutamic acids are bound.

Folic acid may be extracted or purified from food, or may be chemically synthesized. Foods containing folic acid are not particularly limited, but include vegetables such as asparagus, broccoli, spinach, edamame, broad bean, corn, Brussels sprouts, kale, Chinese garlic, and chrysanthemum, and lychee, strawberry, mango, avocado, durian, and the like. Mention may be made of fruits and meats such as chicken, cow and pig liver. In addition, the method for synthesizing folic acid should be based on a known method (8th Edition Food Additives Official Manual, pages D-1655 to D-1660, Hirokawa Shoten Co., Ltd., published on December 10, 2007). For example, while maintaining an equimolar aqueous solution of 2,4,5-triamino-6-hydroxypyrimidine and valaaminobenzoylglutamic acid at pH 4, an ethanol solution of α,β-dibromopropionaldehyde is added to condense it. Purified folic acid can be obtained by dissolving in an aqueous solution of pH 9 or more, adjusting the pH to 7, and removing insoluble matter (this method is not limited).

The composition of the present invention may optionally contain folic acid in an amount selected from the range of 0.1 ppm to 10 ppm, preferably 1 ppm to 10 ppm, more preferably 1 ppm to 5 ppm. For example, the composition of the present invention contains folic acid of 100 μg or more, 150 μg or more, 200 μg or more, 240 μg or more, 250 μg or more, 300 μg or more, 350 μg or more, 400 μg or more, 450 μg or more, 500 μg or more, 550 μg or more per oral intake. It can be appropriately included within the above range. For example, a range that includes the intake amount (day) of folic acid recommended in the “Dietary Reference Intakes for Japanese (2015 edition)” published by the Ministry of Health, Labor and Welfare is preferable. The term "single oral intake" refers to the amount of the composition that is orally ingested at one time, or multiple doses continuously at short time intervals (for example, 10 minutes or less, preferably 5 minutes or less). It means the total amount taken orally. When the composition is in a liquid or semi-solid (gel, sol, etc.) form, for example, 50 mL to 500 mL (typically 50 mL, 100 mL, 150 mL, 180 mL, 200 mL, 250 mL, 300 mL, 350 mL, 400 mL, 450 mL or 500 mL) is the amount. The upper limit of the amount of folic acid contained in one oral intake is not particularly limited, and can be appropriately determined from the range of, for example, 1000 μg or less, 900 μg or less, 800 μg or less, 700 μg or less, or 600 μg or less.

In the present invention, the term "protic organic solvent" means an organic solvent that dissociates by itself to generate protons, and has atoms with high electronegativity, i.e., hydrogen atoms bonded to nitrogen atoms or oxygen atoms. means Examples of such protic organic solvents include, but are not limited to, lower alcohols (ethanol, methanol), glycerin, acetic acid, formic acid, butylamine, and the like.

As detailed in the examples below, the protic organic solvent has the effect of increasing the rate of decomposition of folic acid in an acidic composition. Therefore, by reducing the amount of the protic organic solvent contained in the acidic composition, the rate of decomposition of folic acid in the acidic composition can be reduced, and the storage stability of folic acid in the acidic composition can be enhanced. can be done.

The amount of the protic organic solvent contained in the composition of the present invention is preferably reduced as much as possible in order to reduce the rate of decomposition of folic acid and/or enhance the storage stability. The storage stability of folic acid in the composition can be enhanced by reducing the content of the protic organic solvent. For example, by reducing the amount of protic organic solvent included relative to conventional acidic compositions containing folic acid, the rate of folic acid degradation is reduced and/or storage is reduced relative to conventional acidic compositions. A highly stable composition can be obtained. Therefore, the amount of the protic organic solvent contained in the composition of the present invention may be less than the amount of the protic organic solvent in conventional acidic compositions containing folic acid, and is not particularly limited. but can range from less than 0.1 wt%, such as less than 0.09 wt%, less than 0.08 wt%, less than 0.07 wt%, or less than 0.06 wt%. Particularly preferably, it can be in the range of 0.05% by weight or less, 0.04% by weight or less, 0.03% by weight or less, 0.02% by weight or less, or 0.01% by weight or less.

The "protic organic solvent" in the present invention includes not only the protic organic solvent blended in the manufacturing process of the acidic composition of the present invention, but also the raw material used in manufacturing the composition. Contaminating protic organic solvents are also included. For example, ethanol is contained in almost all kinds of perfume preparations today (Uematsu et al., Shokueishi, Vol.38, No.6, pp.452-459, December, 1997). However, for commercially available fragrance formulations, although the diluent used in the final product is labeled, the solvent used for extracting the fragrance may not be labeled. Since the fragrance is mixed, the solvent contained in the fragrance before mixing may not be displayed. Therefore, even if there is no indication that the fragrance formulation contains a protic organic solvent, it may actually contain the solvent. Increasing the content of protic organic solvents in the composition can increase the rate of folic acid degradation.

The amount of protic organic solvent in the composition of the present invention can be measured by a conventionally known general method. For example, the content of the protic organic solvent in the composition is measured by mixing the composition with anhydrous sodium sulfate and acetonitrile, filtering this, and dissolving the obtained filtrate in acetonitrile as an analysis sample. (Kenji Isshiki, Shokuei Shi. Vol. 26, No. 1, pp. 39-45, 1985).

In the present invention, "acidic composition" means a composition having a pH value of 4.5 or less, for example, pH less than 4.5, pH less than 4.4, pH less than pH 4.3, pH less than pH 4.2, pH 4.5 or less. less than .1, less than pH 4.0, less than pH 3.9, less than pH 3.8, less than pH 3.7, less than pH 3.6, less than pH 3.5, less than pH 3.4, or less than pH 3.3 . The lower limit of the pH value is not particularly limited, and can be determined appropriately according to the form of the acidic composition. .9 or higher, pH 3.0 or higher, pH 3.1 or higher, or pH 3.2 or higher.

When the acidic composition of the present invention is in the form of a soft drink, the pH value is preferably less than pH 4.0. According to the Food Sanitation Act's standards for each food product, the methods of sterilizing and removing bacteria from soft drinks differ greatly at pH 4.0. The sterilization of pH 4.0 or higher requires heating at a core temperature of 85° C. for 30 minutes. Therefore, by setting the upper limit of the pH value to less than pH 4.0, the burden of the sterilization process can be reduced, and the decomposition of folic acid in the sterilization process can be reduced, which is preferable. In addition, in this specification, a pH value points out the value measured by the product temperature of 20 degreeC.

The pH value of the acidic composition of the present invention can be adjusted by adjusting the amount of acidulant added accordingly. Examples of acidulants include those commonly used in the production of pharmaceuticals and food and beverages, such as citric acid, malic acid, gluconic acid, tartaric acid, lactic acid, succinic acid, and salts thereof. can be added, one or a mixture of two or more of

In addition to folic acid, the acidic composition of the present invention contains excipients, disintegrants, lubricants, binders, antioxidants, coloring agents, anti-aggregating agents, and absorption enhancers that are acceptable as pharmaceuticals or foods. , Solvents, solubilizers, tonicity agents, stabilizers, flavoring agents, pH adjusters, fragrance preparations, sweeteners, thickeners, preservatives, vitamins, agar, etc. They can be appropriately selected and blended depending on the desired form of the product. However, it is preferable to use those components that do not contain a protic organic solvent or that have a low protic organic solvent content. For example, as mentioned above, today almost all types of perfume formulations contain ethanol as a solvent. In the present invention, instead of such a perfume preparation, a perfume preparation using one or more selected from medium-chain fatty acids and/or emulsifiers instead of ethanol as a solvent can be included. Although not particularly limited here, medium-chain fatty acids that can be used include C8, C10, and C12 triglycerides, and emulsifiers that can be used include glycerin fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, A sucrose fatty acid ester or the like can be used.

The acidic composition of the present invention can be provided in the form of pharmaceuticals (including quasi-drugs) or food and drink. These forms can be in the form of liquid compositions or semi-solid compositions (gel, sol, etc.), etc. In addition to the folic acid, the other raw materials are appropriately added according to the dosage form. can be blended and prepared according to conventional methods.

For example, the acidic compositions of the present invention can be packaged beverages. The container for containing the acidic composition (liquid composition) of the present invention may be a container used as a container for beverages, and is not limited thereto. , metal can containers, and the like. The shape of the container is not particularly limited. In addition, although the volume of the container is not particularly limited, it is, for example, 50 to 500 mL (typically 50 mL, 100 mL, 150 mL, 180 mL, 200 mL, 250 mL, 300 mL, 350 mL, 400 mL, 450 mL or 500 mL), preferably 100 to 200 mL. be able to.

Alternatively, the acidic composition of the present invention can be a packaged jelly drink. The container for containing the acidic composition (semi-solid (gel, sol, etc.) composition) of the present invention may be a container used as a container for jelly beverages, and is not limited thereto. Examples include films and/or metal film containers, pouch containers, and the like. The capacity of the container is not particularly limited, but can be, for example, 50 mL to 200 mL (typically 50 mL, 100 mL, 150 mL, 180 mL, 200 mL).

The acidic composition of the present invention is obtained by mixing folic acid extracted or purified from food or chemically synthesized folic acid, an acidulant, other raw materials as described above as necessary, and water as the balance, The amount of protic organic solvent is less than the amount of protic organic solvent in conventional acidic compositions containing folic acid, less than 0.1 wt%, such as less than 0.09 wt%, 0.08 wt% %, less than 0.07 wt%, or less than 0.06 wt%, preferably 0.05 wt% or less, 0.04 wt% or less, 0.03 wt% or less, 0.02 wt% or less , or in the range of 0.01% by weight or less. The method for containing the acidic composition of the present invention in a container and for sterilization can be selected arbitrarily.

EXAMPLES The present invention will be described in more detail below with reference to Examples and Test Examples, but the present invention is not limited to these Examples.

I. Material/measurement method 1. Standard undiluted solution 100 mg of folic acid was precisely weighed in a 200 mL brown beaker, and 5 mL of 0.1N sodium hydroxide solution (Wako Pure Chemical Industries) was added and dissolved. Next, after adding and mixing 10 mL of special grade ethanol (Wako Pure Chemical), the pH of the solution was adjusted to 7.0 to 8.0 with 100 mL of deionized water and 0.1N hydrochloric acid (Wako Pure Chemical). A 200 mL brown volumetric flask and ion-exchanged water were used to make a predetermined volume, which was used as a standard stock solution for preparing a calibration curve in HPLC measurement.

2. HPLC measurement A solution sample containing folic acid was weighed and then adjusted to pH 12.0 (pH meter (F-72) manufactured by HORIBA) using a 1.0N sodium hydroxide solution (Wako Pure Chemical Industries). Then, after stirring for 10 minutes (Magnetic stirrer (RSH-1AN) manufactured by AS ONE), the pH was adjusted to 7.0 using 1.0 N hydrochloric acid (Wako Pure Chemical Industries) and filtered (0.45 μm filter manufactured by AS ONE (SY25TF )), and the obtained filtrate was subjected to HPLC measurement.
HPLC measurement was performed under the following conditions.

Apparatus: Waters ACQUITY H-Class system Column: Waters XBridge C18 5 μm 6×250 mm
Temperature: 50°C
Flow rate: 1.2ml/min
Mobile phase: 12% methanol (Wako Pure Chemical), 0.3% acetic acid (Wako Pure Chemical), 1.08% sodium octasulfonate solution (Wako Pure Chemical)

3. Base liquid 2.5% by weight of granulated sugar, 1.2% by weight of fructose, 0.8% by weight of L-ascorbic acid, 0.2% by weight of citric acid, 0.2% by weight of trisodium citrate in ion-exchanged water, and 0.0003% by weight of folic acid was added, and 1.0N hydrochloric acid (Wako Pure Chemical Industries) and 1.0N sodium hydroxide (Wako Pure Chemical Industries) were used to adjust the pH to a predetermined value to prepare a base solution.

II. Test 1: Effect of ethanol addition (1)
According to the composition shown in Table 1 below, solutions containing a base solution and ethanol or acetonitrile and having a predetermined pH (Examples 1 to 7, Comparative Examples 1E to 7E (ethanol added), Comparative Examples 1A to 7A (Acetonitrile addition)) were prepared respectively (day 0).

100 mL of each solution was filled into a light-shielded retort pouch at room temperature, and then sterilized with hot water at 80° C. for 10 minutes (constant temperature water bath (D20KP) manufactured by LAUDA). Then, each solution was stored at 50° C. for a predetermined period (ADVANTEC thermostat (AGX-345)), and the amount of folic acid in each solution after storage was measured by HPLC.

Table 2 below shows the measurement results of the amount of folic acid in each solution after storage. In addition, the numerical value of the amount of folic acid in the table is shown as a relative value with the amount of folic acid in the solution on the 0th day being set to "100".

As shown in Table 2, it was confirmed that in Comparative Examples 1E to 7E in which the proton-donating organic solvent ethanol was added, the folic acid decomposition rate was significantly increased.

On the other hand, in Comparative Examples 1A to 7A with the addition of acetonitrile (SP value, ethanol: 12.7, acetonitrile: 11.9) having a dissolution parameter (SP value) similar to that of ethanol, folic acid similar to that of the example Amounts were confirmed and the addition of acetonitrile was not found to significantly increase the rate of folic acid degradation.

These results indicate that the increased rate of degradation of folic acid observed in Comparative Examples 1E-7E is not due to the solubility of folic acid in the solvent, but due to the presence of ethanol upon addition of ethanol. .

III. Test 2: Effect of ethanol addition (2)
A solution (Examples 8 and 9, Comparative Example 8E) containing a base solution and ethanol and having a pH of 3.5 was prepared according to the composition shown in Table 3 below (Day 0).

Each solution was filled in a retort pouch, sterilized and stored in the same manner as in Test 1 above, and the amount of folic acid in each solution after storage was measured by HPLC.

Table 4 below shows the measurement results of the amount of folic acid in each solution after storage. In addition, the numerical value of the amount of folic acid in the table is shown as a relative value with the amount of folic acid in the solution on the 0th day being set to "100".

As shown in Table 4, the addition of ethanol reduced the amount of folic acid after storage (compared with Example 4 in Table 2 above), but the examples containing 0.03% by weight and 0.05% by weight of ethanol Compared to 8 and 9, Comparative Example 8E containing 0.5% by weight of ethanol was confirmed to have a significantly lower amount of folic acid after storage. On the other hand, between Examples 8 and 9, no significant difference was observed in the amount of folic acid after storage.

This result confirms that the higher the ethanol content, the higher the folic acid decomposition rate, but it is suggested that when the ethanol content is 0.05% by weight or less, the folic acid decomposition rate does not change much. be done.

IV. Test 3: Effect of ethanol addition (3)
Solutions containing folic acid (Example 10, Comparative Example 9E) were prepared according to the compositions shown in Table 5 below (Day 0). Fragrance 1 is a fragrance obtained by using a medium-chain fatty acid and a glycerin fatty acid ester as a solvent without containing ethanol, and is a fragrance made into a solution, and fragrance 2 contains 50% by weight of ethanol as a solvent (final product concentration 0.1% by weight). It is perfume.

Each solution was sterilized and stored in the same manner as in Test 1 above, and the amount of folic acid in each solution after storage was measured by HPLC.

Table 6 below shows the measurement results of the amount of folic acid in each solution after storage. In addition, the numerical value of the amount of folic acid in the table is shown as a relative value with the amount of folic acid in the solution on the 0th day being set to "100".

As shown in Table 6, it was confirmed that the addition of ethanol increased the decomposition rate of folic acid in the solution, even in the form of being contained in perfume.

Generally, in beverages and water-based foods, ethanol-containing preparations are used as fragrances in order to improve dispersibility. However, the above results show that it is advantageous to use ethanol-free fragrances, that is, to use ethanol-free raw materials in order to reduce the rate of decomposition of folic acid and stabilize it. rice field.

Claims (4)

An acidic composition having a pH of 4.5 or less, containing folic acid and a perfume preparation containing the solvent and not containing ethanol, which contains a perfume produced using one or more selected from medium-chain fatty acid triglycerides and emulsifiers as a solvent. (excluding acidic compositions containing folic acid and caffeine), characterized in that the content of ethanol is less than 0.1% by weight. The composition according to claim 1, which is a packaged beverage or a packaged jelly beverage. An acidic composition having a pH of 4.5 or less, containing folic acid and a perfume preparation containing the solvent and not containing ethanol, which contains a perfume produced using one or more selected from medium-chain fatty acid triglycerides and emulsifiers as a solvent. (excluding an acidic composition containing folic acid and caffeine), comprising blending raw materials such that the blending amount of ethanol is less than 0.1% by weight. 4. The method of claim 3, wherein the acidic composition is a packaged beverage or a packaged jelly beverage.