JP6858675B2 - Foods and drinks containing amperopsin and polyglutamic acid - Google Patents

Description

The present invention relates to foods and drinks containing ampelopsin and polyglutamic acid.

Amperopsin is a flavonol compound contained in wisteria tea.
Wisteria tea, a plant containing ampelopsis, is classified into the genus Ampelopsis of the Grapes family, which is mainly produced in China, and its Chinese name is Akito snake grape. The scientific name is Ampelopsis grossedentata. This plant is mainly distributed in provinces and autonomous regions such as Guangxi, Guangxi, Yunnan, Guizhou, Hunan, Hubei, Jiangxi and Fujian in China. Beverages made from these stems and leaves are commonly used as tea by the Zhuang and Yao people in provinces and autonomous regions such as Guangxi and Hunan in China (this is called "Fujicha"), and colds and throats. It is also used for pain. Amperopsin has various physiological and pharmacological activities exhibited by Fujicha, such as therapeutic action for liver disease (Patent Document 1), lipase inhibitory action (Patent Document 2), matrix protease inhibitory action (Patent Document 3), and antibacterial action (Patent Document 3). It is specified as the main body of action and activity such as 4), deterioration prevention action of fragrance (Patent Document 5), and fading prevention action of dye (Patent Document 6). Further, the applicant has found an action of suppressing salt-dependent hypertension generated by salt intake, and has already applied for a patent application (Japanese Patent Application No. 2017-053146) as a composition for improving hypertension.

The amount of ampelopsin contained in the above-mentioned beverage (beverage containing an extract of wisteria tea) decreases rapidly with the passage of time. That is, ampelopsin is extremely unstable when dissolved or dispersed in water. The present inventors have confirmed this fact by previous studies, and it can be said that ampelopsin is an extremely unstable substance in the presence of water. Therefore, in order to make amperopsin a food and drink that can be ingested in a stable state, it must be extracted from the above wisteria tea at the time of eating and drinking and drunk promptly, or water should be removed promptly after extraction to make it a dry product, or blood pressure. The lowering effect cannot be expected.
It is clear that such instability of ampelopsin hinders the spread of foods and drinks containing wisteria tea and wisteria tea extract. However, a technique for stabilizing ampelopsin in an aqueous solution has not yet been provided.

On the other hand, the above-mentioned action of ampelopsin to suppress salt-dependent hypertension can be further enhanced by reducing the daily amount of salt ingested by meals. However, reducing the salt content of a diet reduces the saltiness of the food, which is directly linked to a decrease in appetite. Therefore, it is necessary to improve eating habits and it is actually difficult to reduce the intake of salt derived from the diet. Therefore, for the purpose of reducing the salt content in the diet without lowering the salty taste, a method of using potassium chloride instead of salt, a method of adding various amino acids (Patent Document 7), and polyglutamic acid are added. A method (Patent Document 8) and the like have been proposed.

Japanese Unexamined Patent Publication No. 2003-26584 Japanese Unexamined Patent Publication No. 2003-12536 JP-A-2002-173424 JP-A-2002-159566 Japanese Unexamined Patent Publication No. 2004-18756 JP-A-2002-65201 JP-A-2002-345430 International Publication No. 2007/108558

The present inventors have studied foods and drinks containing polyglutamic acid and ampelopsin as foods and drinks having a blood pressure lowering effect while satisfying the desire for salty taste. In the process, it was found that both ampelopsin and polyglutamic acid became unstable in the aqueous solution, and in particular, amperopsin decreased at a higher rate as the concentration of polyglutamic acid increased. Therefore, a technique for stabilizing ampelopsin was investigated.

That is, an object of the present invention is to provide a food or drink composition containing ampelopsin and polyglutamic acid and stabilizing ampelopsin.

The main configuration of the present invention is as follows.
(1) Contains ampelopsin, polyglutamic acid and an organic acid, and contains 0.005 to 1% by mass of ampelopsin, 0.01 to 3% by mass of polyglutamic acid and 0.003 to 0.3% by mass of an organic acid per composition. Food and beverage composition.
(2) The food or drink composition according to (1), wherein the organic acid is one or more substances selected from gluconic acid, lactic acid, succinic acid, malic acid, tartaric acid, and phytic acid.
(3) The food or drink composition according to (1) or (2), which has a pH of 2.5 to 5.
(4) The composition according to any one of (1) to (3), wherein ampelopsin is derived from a wisteria tea extract.

INDUSTRIAL APPLICABILITY The present invention provides a stable food or drink containing ampelopsin and polyglutamic acid. Food and beverage compositions containing one or more substances selected from gluconic acid, lactic acid, succinic acid, malic acid, tartaric acid, and phytic acid as organic acids for stabilizing amperopsin include soft drinks, dairy drinks, and lactic acid bacteria. It can be applied to beverages containing a large amount of water such as beverages, green tea beverages, black tea beverages, fruit juice beverages, vegetable beverages, fermented milk, jelly, ice cream, and ice cream. In particular, it has a favorable flavor as a tea beverage such as green tea beverage and black tea beverage, and ampelopsin does not decrease even after long-term storage.
In addition, since the composition of the present invention contains polyglutamic acid, when it is drunk at the time of meal, it gives a favorable salty taste to the food to be ingested, so that the salt content of the food can be reduced. In addition, by taking it at the time of eating and drinking, it is possible to control blood pressure normally by acting synergistically with the blood pressure suppressing effect of ampelopsin.

The present invention contains ampelopsin, polyglutamic acid and an organic acid, and contains 0.005 to 1% by mass of ampelopsin, 0.01 to 3% by mass of polyglutamic acid and 0.003 to 0.3% by mass of an organic acid per composition. It is an invention relating to the food and drink composition.
As mentioned above, wisteria tea is a plant belonging to the genus Ampelopsis in the family Grapes, and its Chinese name is scabbard. The scientific name is Ampelopsis grossedentata. Amperopsin, which is mainly distributed in provinces such as Guangxi, Guangxi, Yunnan, Guizhou, Hunan, Hubei, Jiangxi, and Fujian, and autonomous regions of China, is a compound represented by the following formula.

AMPelopsin, for example, Fujicha (Ampelopsis grossedentata), DaiKano snake grape (Ampelopsis megalophylla), Guangdong snake grape (Ampelopsis cantoniensis), Kenponashi (Hovenia dulcis), Onoeyanagi (Salix sachalinensis), Yorehamatsu (Pinus contorta), Erythrophleum africanum and It can be isolated and purified from an extract of a plant selected from Katsura (Cerciphysyllum japonicum). Among these, wisteria tea is preferable.
Specifically, ampelopsis can be obtained from Ampelopsis planta, which is a plant belonging to the genus Ampelopsis, as follows.
That is, the extract obtained by extracting the branches and leaves of dried wisteria tea with hydrous ethanol is concentrated, subjected to column chromatography using, for example, a porous resin (DIAION HP-20), and eluted with 80% by volume hydrous methanol. Amperopsin is obtained in the picture. This can be further purified by reverse phase silica gel column chromatography or recrystallization. The purified ampelopsin is also sold as a reagent, and this can also be used.

For food and drink compositions such as health foods, a commercially available extract containing 10% by mass or more of the above ampelopsin can be used. In order to obtain such an extract from wisteria tea, the following operations are performed.
Using crushed dried wisteria tea leaves or stems or powder as an extraction raw material, it is put into water or a hydrophilic organic solvent or a mixed solvent thereof, and extracted using an arbitrary device at room temperature or a temperature below the boiling point of the solvent. Can be obtained by

Examples of the organic solvent used for extraction include lower alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene glycol, propylene glycol and iso. Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as propylene glycol and glycerin.
Further, a mixed solvent of the above-mentioned hydrophilic organic solvent and water can be used. When a mixed solvent of water and a hydrophilic organic solvent is used, the amount of lower alcohol is 30 to 90 parts by mass with respect to 10 parts by mass of water, and the amount of lower aliphatic ketone is 10 parts by mass of water. On the other hand, it is preferable to add 10 to 40 parts by mass, and in the case of polyhydric alcohol, 10 to 90 parts by mass with respect to 10 parts by mass of water.

A dried and crushed product of wisteria tea is put into a treatment tank filled with an extraction solvent, and the mixture is allowed to stand for 30 minutes to 2 hours with occasional stirring as necessary to elute soluble components, and then filtered to form a solid product. Is removed, the extraction solvent is distilled off from this extract, and the extract is dried to obtain an extract. The amount of the extraction solvent is preferably 5 to 15 times the amount (mass ratio) of the extraction raw material, and the extraction conditions are usually about 1 to 4 hours at 50 to 95 ° C. when water is used as the extraction solvent. Is. When a mixed solvent of water and ethanol is used as the extraction solvent, it is usually about 30 minutes to 4 hours at 40 to 80 ° C.

When the extraction solvent is distilled off from the obtained extract, a paste-like concentrate is obtained. Further drying gives a solid extract. In the present invention, as long as the content of ampelopsin is 10% by mass or more, preferably 20% by mass or more, the above extract or a concentrated solution thereof may be used. These may be used after being purified by a method such as activated carbon treatment, adsorption resin treatment, ion exchange resin, or liquid-liquid countercurrent distribution.
Therefore, an extract extracted from the above-mentioned wisteria tea and having an increased concentration of ampelopsin can also be used as a raw material for foods and drinks of the present invention.

The content of ampelopsin in the composition can be analyzed by a known method such as HPLC. The outline of the quantification method is as follows.
-Preparation of sample solution Approximately 20 mg of a sample (extract) is precisely weighed, distilled water is added, and sonication is performed to dissolve the sample (extract) to make exactly 50 mL. Accurately dilute 2 mL of this solution to 50 mL to make a sample solution.
・ Preparation of standard solution and preparation of calibration curve
Weigh 2.00 mg of a standard product (manufactured by Sigma-ALdrich), add an appropriate amount of 100% acetonitrile to dissolve it, and then add acetonitrile to make exactly 25 mL, and add 80 μg / mL of ampelopsin standard stock solution. Prepare. This standard stock solution is diluted exactly 5-fold with distilled water to prepare a 16 μg / mL amperopsin standard solution. The injection volume to HPLC is 10, 20, 40 μL, and a calibration curve is prepared based on the peak of ampelopsin. This measuring method can also be used for quantifying amperopsin contained in foods and drinks.
-HPLC measurement conditions Measurement can be performed under the standard measurement conditions shown in Table 1 below.

Amperopsin has a chimase inhibitory effect and suppresses the increase in blood pressure caused by the renin-angiotensin system.
Chymotrypsin is a chymotrypsin-like enzyme produced and secreted by mast cells. It has been known that human blood vessels have an angiotensin (Ang) II production mechanism that does not belong to the conventional renin-angiotensin (RA) system. In recent years, it has been found that the serine protease involved in this production mechanism is chimase. AngII has recently attracted attention as a factor involved in pathological tissue remodeling, which is widely regarded as an organ disorder, as well as blood pressure regulation in the circulatory system.
By the action of renin, angiotensinogen produces angiotensin I, which consists of 10 amino acid residues. Then, the two residues at the C-terminal are cleaved by the action of angiotensin converting enzyme (ACE), chimase, and cathepsin G, and converted to angiotensin II (AngII). Human chimase is believed to be a specific AngII-producing enzyme that acts in human tissues rather than in the circulating blood.

Kimase activity inhibitory activity (in vitro) can be confirmed by testing by the following method, which is a modification of the method described in Eur J Biochem 268 (22), 5885-5893 (2001).
This evaluation method is a simple method using a substrate obtained by modifying angiotensin I, which is a substrate of human chimase, with Dnp / Nma, and when the chimase cleaves this substrate and produces angiotensin II, fluorescent color is developed. The outline of the evaluation method is described below.
The incubation buffer is a 20 mM phosphate buffer containing 100 mM NaCl, and the total amount of the incubation solution is 100 μL. First, 5 μL of a sample was added, adjusted so that 0.0012 units of standard human chymase (manufactured by SIGMA-ALDRICH) were contained therein, preincubation was performed at room temperature for 30 minutes, and then the substrate Dnp / Nma-modified angiotensin I was added. Add to a final concentration of 200 μM and incubate at 37 ° C. for 30 minutes. Add 25 μL of 0.5 M NaOH to complete the incubation. The emission fluorescence (460 nm) of the produced Dnp angiotensin II is measured, and the production amount is calculated from the standard curve prepared by the standard Dnp angiotensin II. Using the control to which the test sample is not added as a control, the inhibition of human chimase activity and the inhibition rate of the test sample are determined.
Purified ampelopsin suppresses chymase activity 100% at a concentration of 200 μM.

In addition, the effect of improving hypertension can be confirmed by orally administering 2% saline solution to a model animal (mouse) having hypertension by giving it in a state of free water intake for 12 weeks. Ampelopsin is administered at 200 mg / kg (body weight) to improve salt-induced hypertension.

The above-mentioned wisteria tea extract is added to the food and drink composition of the present invention as it is or diluted with water or the like. In order to exhibit the pharmacological or physiological effect of ampelopsin, 0.005 to 1% by mass of ampelopsin is added to the food or drink. If it is less than 0.005% by mass, a pharmacological effect such as a hypertension improving effect or a physiological effect cannot be expected, and if it exceeds 1% by mass, the astringent taste and bitterness peculiar to amperopsin become stronger.

Further, in the present invention, 0.01 to 3% by mass of polyglutamic acid and 0.003 to 0.3% by mass of organic acid are blended. If the concentration of polyglutamic acid is less than 0.01% by mass, there is no salty taste reinforcing effect on the food, and if it exceeds 3% by mass, the flavor as a food or drink is affected, which is not preferable. The food and drink of the present invention is adjusted to pH 2.5 to 6 using an acid or alkaline aqueous solution. When the pH exceeds 6, ampelopsin decreases over time.
Polyglutamic acid (Poly-γ-Glutamic Acid) is an amino acid polymer (polymer) in which D-glutamic acid and L-glutamic acid are mixed, and its DL ratio is about 8: 2. This substance is characterized by having a large number of negative ions, and natto Neva is a polymer in which 30 to 5000 pieces of this glutamic acid are bound. As the polyglutamic acid used in the present invention, γ-polyglutamic acid in the mucilage of natto may be extracted and used, or γ-polyglutamic acid secreted outside the cells of the genus Bacillus such as Bacillus natto may be used. In addition, there is no problem even if levan is contained in the natto mucilage or simultaneously secreted by Bacillus natto. Polyglutamic acid can be produced by chemical synthesis, enzymatic synthesis, or culture using cells of the genus Bacillus such as Bacillus natto. The technique for producing from Bacillus natto is known, and it may be produced by the method disclosed in International Publication No. 2003/049771. When Bacillus natto or the like is used, polyglutamic acid contained in the mucilage of natto can be extracted, or polyglutamic acid secreted by the cells to the outside of the cells can be used.
In general, polyglutamic acid in natto mucilage and polyglutamic acid secreted by the genus Bacillus under normal culture conditions are high molecular weight substances and extremely viscous. be able to.
In addition, polyglutamic acid has a calcium absorption promoting action and is commercially available as a food additive for promoting calcium absorption. This can also be used. As a commercial product, the trade name "Cultake" manufactured by Ajinomoto Co., Inc. can be exemplified.

The organic acid used in the present invention may be any organic acid that can be blended in foods and drinks. Examples thereof include itaconic acid, citraconic acid, citric acid, acetic acid, succinic acid, gluconic acid, lactic acid, malic acid, tartaric acid, fumaric acid, phthalic acid, oxalic acid, and phytic acid. Preferably, the organic acid is a substance selected from gluconic acid, lactic acid, succinic acid, malic acid, tartaric acid, phytic acid, or a mixture of one or more of these acids.

Examples of foods and drinks to which the technique of the present invention can be applied include soft drinks, milk drinks, lactic acid bacteria drinks, green tea drinks, black tea drinks, fruit juice drinks, vegetable drinks, fermented milk, jelly, ice cream, frozen desserts, and the like. It is not always limited to these.
Other food additives, fruit juices, and the like can be used in the food and drink composition of the present invention as long as the object of the present invention is not impaired. Specifically, dietary fibers and thickeners such as hemicellulose, lignin, guagam, konjakmannan, isagol, alginic acid, agar, carrageenan, chitin, carboxylmethylcellulose, polydextrose, cooking oil, calcium, iron, sodium, zinc. , Copper, potassium, phosphorus, magnesium, iodine, manganese, selenium and other minerals, vitamin A, vitamin C, vitamin D, vitamin E, vitamin K, niacin, folic acid, pantothenic acid and other fat-soluble or water-soluble vitamins , Glycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, phospholipid, arabic gum, xanthan gum, tragacanth gum, locust bean gum and other emulsifiers and dispersants, bulking agents, excipients, preservatives / oxidation Inhibitors, flavor modifiers and fragrances, flavoring agents such as sodium chloride, sodium glutamate, glycine, multitoll, low-calorie sweeteners such as aspartame, coloring agents, etc.

The present invention will be further described below with reference to production examples of ampelopsin and test examples using the ampelopsin.
<Production example 1 of wisteria tea extract>
15 times the amount of water was added to 1 part by weight (200 g) of dried wisteria tea, heated to 90 ° C., extracted for 1 hour and filtered to obtain the first extract. Next, 12 times the amount of water was added to this residue, the mixture was heated to 90 ° C., and then filtered to obtain a second extract. Both extracts were combined, concentrated under reduced pressure, and about 300 mL of the concentrate was frozen at −40 ° C. and further dried in a freeze-dryer to obtain a dried product. This was pulverized and sieved with a 60-mesh sieve to obtain 85 g of a passing product. The ampelopsin content in this composition was 65.8% by mass.

<Production example 2 of wisteria tea extract>
A 50% ethanol aqueous solution was added in an amount of 15 times to 1 part by weight (200 g) of dried wisteria tea, heated with a reflux chiller, extracted for 1 hour and filtered to obtain the first extract. Next, 12 times the amount of a 50% ethanol aqueous solution was added to this residue in the same manner, the mixture was heated to 90 ° C., extracted for 30 minutes and filtered to obtain a second extract. Both extracts were combined, concentrated under reduced pressure, and about 300 mL of the concentrate was frozen at −40 ° C. and further dried in a freeze-dryer to obtain a dried product. This was pulverized and sieved with a 60-mesh sieve to obtain 78.1 g of a passing product. The ampelopsin content in this composition was 52.9% by mass.

<Production example 3 of wisteria tea extract>
In the same manner as in Production Example 1, 15 times the amount of water was added to 1 part by weight of dried wisteria tea, heated to 90 ° C., extracted for 1 hour, and filtered to obtain the first extract. Next, 12 times the amount of water was added to this residue, the mixture was heated to 90 ° C., extracted for 30 minutes and filtered to obtain a second extract. Both extracts were added up and concentrated under reduced pressure to obtain about 300 mL of the concentrate. Further, 78 g of the guar gum decomposition product was added to the concentrate, frozen at −40 ° C., and further dried by a freeze-drying device to obtain a dried product. This was pulverized and sieved with a 60-mesh sieve to obtain 163.1 g of a passing product. The ampelopsin content in this composition was 24.7% by mass.

<Production example 4 of wisteria tea extract>
In the same manner as in Production Example 1, 15 times the amount of water was added to 1 part by weight of dried wisteria tea, heated to 90 ° C., extracted for 1 hour, and filtered to obtain the first extract. Next, 12 times the amount of water was added to this residue, the mixture was heated to 90 ° C., extracted for 30 minutes and filtered to obtain a second extract. Both extracts were added up and concentrated under reduced pressure to obtain about 300 mL of the concentrate. Further, 78 g of γ-cyclodextrin was added to the concentrated solution, frozen at −40 ° C., and further dried in a freeze-drying device to obtain a dried product. The ampelopsin content in this composition was 26.4% by mass.

<Test Example 1>
Confirmation test of destabilizing effect of ampelopsin by polyglutamic acid 1. Test method An aqueous solution containing 100 ppm (0.1% by mass), 300 ppm (0.3% by mass), and 1000 ppm (1% by mass) of ampelopsin was prepared using a wisteria tea extract having an ampelopsin content of 89.9% by mass. did. Polyglutamic acid (“Cultake” manufactured by Ajinomoto Co., Inc.) was added to this aqueous solution, and the effect of ampelopsin on storage stability was evaluated.
In addition, polyglutamic acid was dissolved in the above-mentioned aqueous solution of ampelopsin so as to have the concentration shown in Table 2. Then, the pH of each test solution was measured. Then, after storing at 60 ° C. for 1 week, the ampelopsin content was analyzed by liquid chromatography.

The measurement by liquid chromatography was carried out according to the following procedure.
(1) Reagents used-Acetonitrile (for HPLC)
・ Distilled water ・ Phosphoric acid (special grade reagent)
-0.1% phosphoric acid aqueous solution-Amperopsin standard product (Amperopsin content ≥ 98%
(Manufactured by HPLC SIGMA-ALDRICH))
(2) HPLC method Column: InertStainC18 Inner diameter 3.0 mm x length 100 mm,
Particle size 3 μm (manufactured by GL Sciences Co., Ltd.)
Mobile phase: Solution A: 0.1% aqueous phosphoric acid solution, Solution B: acetonitrile,
A / B ・ ・ ・ 85/15
Column temperature: 40 ° C
Detection wavelength: UV290nm
Flow velocity: 1.0 mL / min
Injection volume: 2 μL
(3) Concentration of sample and standard solution ・ Preparation of sample solution 1 mL of the sample is dissolved in acetonitrile to make exactly 10 mL, which is used as the sample solution.
-Preparation of standard solution 10 mg of amperopsin standard product was precisely weighed, 100% acetonitrile was added in an appropriate amount and dissolved by ultrasonic treatment, and acetonitrile was added to make exactly 50 mL. After preparing 200 μg / mL of amperopsin standard stock solution, 0. .625, 1.25, 2.5, 5, 10, 20 μg / mL is appropriately diluted with acetonitrile to prepare a standard solution. 2 μL of each of the sample solution and the standard solution was injected into the HPLC under the condition (2), the area of ampelopsin was obtained, the calibration curve (y = bx + a: a = section, b = slope) was obtained from the standard solution, and the ampelopsin content was calculated. did.
2. Test results The test results are shown in Table 2 below.

As shown in Table 2, it was found that the higher the concentration of commercially available polyglutamic acid, the more destabilized amperopsin. It was also found that amperopsin alone decreased by about 30 to 40% in an aqueous solution.

<Test Example 2>
Search for a substance that stabilizes ampelopsin From the results of Test Example 1, an aqueous solution of ampelopsin destabilizes by about 30 to 40%, so a substance that stabilizes this was searched for.
Search test for ampelopsin stabilizer using commercially available ingredients 1. Test method A wisteria tea extract having an ampelopsin content of 89.9% by mass was prepared, and an aqueous solution containing 100 ppm of ampelopsin was prepared using the extract. Various commercially available natural product-derived antioxidants or organic acids selected by conducting a preliminary test in advance were added to this aqueous solution to evaluate the effect of ampelopsin on the storage stability.
The substance to be evaluated was dissolved in an aqueous solution of ampelopsin at the concentrations shown in Table 3. Then, the pH of each test solution was measured. Then, after storing at 60 ° C. for 1 week, the ampelopsin content was analyzed by liquid chromatography.

The measurement by liquid chromatography was carried out according to the following procedure.
(1) Reagents used-Acetonitrile (for HPLC)
・ Distilled water ・ Phosphoric acid (special grade reagent)
-0.1% phosphoric acid aqueous solution-Amperopsin standard product (Amperopsin content ≥ 98%
(Manufactured by HPLC SIGMA-ALDRICH))
(2) HPLC method Column: InertStainC18 Inner diameter 3.0 mm x length 100 mm,
Particle size 3 μm (manufactured by GL Sciences Co., Ltd.)
Mobile phase: Solution A: 0.1% aqueous phosphoric acid solution, Solution B: acetonitrile,
A / B ・ ・ ・ 85/15
Column temperature: 40 ° C
Detection wavelength: UV290nm
Flow velocity: 1.0 mL / min
Injection volume: 2 μL
(3) Concentration of sample and standard solution ・ Preparation of sample solution 1 mL of the sample is dissolved in acetonitrile to make exactly 10 mL, which is used as the sample solution.
-Preparation of standard solution 10 mg of amperopsin standard product was precisely weighed, 100% acetonitrile was added in an appropriate amount and dissolved by ultrasonic treatment, and acetonitrile was added to make exactly 50 mL. After preparing 200 μg / mL of amperopsin standard stock solution, 0. .625, 1.25, 2.5, 5, 10, 20 μg / mL is appropriately diluted with acetonitrile to prepare a standard solution. 2 μL of each of the sample solution and the standard solution was injected into the HPLC under the condition (2), the area of ampelopsin was obtained, the calibration curve (y = bx + a: a = section, b = slope) was obtained from the standard solution, and the ampelopsin content was calculated. did.

2. Test results The test results are shown in Table 3 below.

As shown in Table 3, commercially available natural antioxidants were not very effective, and high stability effects were observed for phytic acid, tartaric acid, and malic acid.
From this test result, it was clarified that the organic acid has an action of stabilizing ampelopsin.

<Test Example 3>
Search test for substances that stabilize an aqueous solution containing ampelopsin and polyglutamic acid From the results of Test Example 1 and Test Example 2, the stabilizing action of an aqueous solution containing ampelopsin and polyglutamic acid was evaluated using an organic acid.
1. 1. Test method An aqueous solution containing 300 ppm (0.03% by mass) of ampelopsin was prepared using a wisteria tea extract having an ampelopsin content of 89.9% by mass. A solution was prepared by adding polyglutamic acid (“Cultake” manufactured by Ajinomoto Co., Inc.) to this aqueous solution so as to be 0.1% by mass. To this aqueous solution, 0.1% by mass of vitamin C, 0.05% by mass of citric acid, 0.1% by mass of gluconic acid, 0.05% by mass of fermented lactic acid, and 0.1% by mass of succinic acid are added to stabilize the storage of ampelopsin. The effect on sex was evaluated.
After measuring the pH of each test solution, it was stored at 60 ° C. for 1 week, and then the ampelopsin content was analyzed by liquid chromatography.

The measurement by liquid chromatography was carried out according to the following procedure.
(1) Reagents used-Acetonitrile (for HPLC)
・ Distilled water ・ Phosphoric acid (special grade reagent)
-0.1% phosphoric acid aqueous solution-Amperopsin standard product (Amperopsin content ≥ 98%
(Manufactured by HPLC SIGMA-ALDRICH))
(2) HPLC method Column: InertStainC18 Inner diameter 3.0 mm x length 100 mm,
Particle size 3 μm (manufactured by GL Sciences Co., Ltd.)
Mobile phase: Solution A: 0.1% aqueous phosphoric acid solution, Solution B: acetonitrile,
A / B ・ ・ ・ 85/15
Column temperature: 40 ° C
Detection wavelength: UV290nm
Flow velocity: 1.0 mL / min
Injection volume: 2 μL
(3) Concentration of sample and standard solution ・ Preparation of sample solution 1 mL of the sample is dissolved in acetonitrile to make exactly 10 mL, which is used as the sample solution.
-Preparation of standard solution 10 mg of amperopsin standard product was precisely weighed, 100% acetonitrile was added in an appropriate amount and dissolved by ultrasonic treatment, and acetonitrile was added to make exactly 50 mL. After preparing 200 μg / mL of amperopsin standard stock solution, 0. .625, 1.25, 2.5, 5, 10, 20 μg / mL is appropriately diluted with acetonitrile to prepare a standard solution. 2 μL of each of the sample solution and the standard solution was injected into the HPLC under the condition (2), the area of ampelopsin was obtained, the calibration curve (y = bx + a: a = section, b = slope) was obtained from the standard solution, and the ampelopsin content was calculated. did.

2. Test results The test results are shown in Table 4 below.

As shown in Table 4, 0.1% by mass of gluconic acid, 0.05% by mass of fermented lactic acid, 0.1% by mass of succinic acid, 0.05% by mass of malic acid and 0.1 in an aqueous solution containing amperopsin and polyglutamic acid. %, Tartaric acid 0.05% by mass / 0.1% by mass, Phytic acid 0.05% by mass / 0.1% by mass, all of which are stored at 10% or more as compared with the control-free test solution. Improved stabilization of ampelopsin in the study.

From the results of Test Examples 1 to 3 above, when ampelopsin coexists with water, the ampelopsin is extremely unstable and rapidly decreases, and when polyglutamic acid coexists, the decrease accelerates, and this ampelopsin It was confirmed that amperopsin was stabilized when an organic acid was added to an aqueous solution containing polyglutamic acid and the pH of the solution was set to 5 or less.
Therefore, it was found that it is effective to add an organic acid to the composition containing ampelopsin and polyglutamic acid to bring the pH to 5 or less.

<Beverage production example>

300 g of water was added to 6 g of dried wisteria tea leaves, and the mixture was extracted at 100 ° C. (boiling) for 15 minutes.
This was cooled and then filtered to obtain a wisteria tea extract.
As shown in Table 5, wisteria tea extract powder, fermented lactic acid, and polyglutamic acid were added to and dissolved in this wisteria tea extract. Finally, water was added to make the total amount 100 g to make a tea beverage.
The pH of this tea beverage was 3.96, and it had a favorable flavor when drunk, making it a highly palatable product.

Claims (4)

Food and beverage composition containing ampelopsin, polyglutamic acid and organic acid, and containing 0.005 to 1% by mass of ampelopsin, 0.01 to 3% by mass of polyglutamic acid and 0.003 to 0.3% by mass of organic acid per composition. Stuff. The food or drink composition according to claim 1, wherein the organic acid is one or more substances selected from gluconic acid, lactic acid, succinic acid, malic acid, tartaric acid, and phytic acid. The food or drink composition according to claim 1 or 2, wherein the pH is 2.5 to 5. The composition according to any one of claims 1 to 3, wherein the ampelopsin is derived from the wisteria tea extract.