JP5459699B2 - Cranberry extract and method for producing the same - Google Patents

Description

  The present invention relates to a cranberry extract that is rich in proanthocyanidin A type and does not contain a large amount of fat-soluble components, and has no unpleasant taste such as astringency and astringency, and can be applied to foods.

  Proanthocyanidins are a compound group in which condensed tannins present in various plant bodies or non-hydrolyzable tannins, that is, flavan-3-ols or flavan-3,4-diols are combined as constituent units by condensation and polymerization. Proanthocyanidins are classified into several types based on the difference in binding mode. For example, two or more flavan-3-ol structures are in the 4β position → 8 position and 2β position → O → 7 position or 4β position → 6 position and 2β. If there is at least one site that binds via the position → O → 7, the A-type, two or more flavan-3-ol structures bind via the 4β position → 8 position or the 4β position → 6 position. It is known that it is classified into B type. Proanthocyanidin type B has been recognized to have various functions and physiological activities, such as hair growth (see, for example, Patent Document 1), antiviral effect (for example, see, Patent Document 2), arteriosclerosis and diabetes. Effects such as prevention of metabolic syndrome represented by (see, for example, Patent Document 3) are known. On the other hand, proanthocyanidin A type inhibits hyaluronidase activity, anti-allergy or anti-inflammatory action (for example, see Patent Document 4), and prevents adhesion of bacteria that cause urinary tract infections (for example, see Non-Patent Document 1). However, it has not been verified in detail. In the background, proanthocyanidin type B is widely contained in various foods such as vegetables and fruits, while proanthocyanidin type A is very limited to cranberries, plums, avocados, peanuts, cinnamon, etc. Another reason is that a method for efficiently extracting proanthocyanidin A type has not yet been established (see Non-Patent Document 2, for example).

  When extracting proanthocyanidins from plant raw materials, generally, a method of extraction using an organic solvent such as water or ethanol, and drying and concentration (for example, see Patent Documents 5, 6, and 7) is known. Although these methods can efficiently extract proanthocyanidin B type from fruit peels and the like, there is a disadvantage that the extraction efficiency of proanthocyanidin A type is poor. In order to compensate for the drawbacks, a method is known in which a solution obtained after extraction is subjected to an adsorption treatment with a synthetic adsorbent or a reverse-phase adsorbent (see, for example, Patent Documents 4 and 11). However, in these methods, when eluting the desired proanthocyanidins from the adsorption resin, a large amount of ethanol or acetone must be used. Along with proanthocyanidins, flavanols represented by quercetin glycosides and the like A large amount of fat-soluble components such as chlorophylls are extracted, synergistically with the astringent taste and astringent taste of proanthocyanidin A type, resulting in an unpleasant taste and the disadvantage that it cannot be used for food. In order to remove the component imparting an unpleasant taste after the extraction of proanthocyanidins, a large amount of organic solvent and a complicated process must be passed.

Journal of Natural Products, 63 (9), 1225-1228 (2000). Journal of Agricultural and Food Chemistry, 51 (25), 7513-7521 (2003).

JP 2004-307370 A JP 2005-314316 A JP 2006-182706 A Japanese Patent No. 2975997 JP-A-9-212484 Table 2005/0779825 gazette Table 2006/068212 Table 2005/030200 Table 2005/072762 JP 2008-156265 A JP 2008-156265 A

  The present invention has an unpleasant taste such as astringency and astringent taste by containing abundant proanthocyanidin A type having an effect of suppressing adhesion of bacteria causing urinary tract infection and not containing a large amount of fat-soluble components. It is an object to provide a cranberry extract that can be applied to foods.

As a result of studying to solve the above-mentioned problems, the present inventor has extracted cranberry peel or pomez with hot water at about 90 ° C. for 2 to 4 hours, thereby producing 1 part by weight of proanthocyanidin B type _2. Knowing that it contains 0.6 parts by weight or more of anthocyanidin A type ₂ and 10% (w / w) or more of a monosaccharide and exhibits a good flavor, the present invention has been completed.

That is, the present invention
1) Proanthocyanidin A type ₂ is contained in an amount of 4% (w / w) or more and 1 part by weight of proanthocyanidin B type ₂ contained in the proanthocyanidin is 0.6 parts by weight or more. A cranberry extract characterized by containing 10% (w / w) or more of a monosaccharide .
2) According to 1) above, 3-5 parts by weight of water is added to 1 part by weight of cranberry peel or cranberry pome, followed by heating, followed by extraction at 80-90 ° C. for 2-5 hours. Method for producing cranberry extract.
Is to provide.

  According to the present invention, it is possible to provide a cranberry extract that can be applied to foods and beverages because flavor is improved by containing abundant monosaccharides and astringency is reduced by not containing a fat-soluble component. .

  Hereinafter, the present invention will be specifically described.

(Proanthocyanidins)
Proanthocyanidins are a compound group in which condensed tannins present in various plant bodies or non-hydrolyzable tannins, that is, flavan-3-ols or flavan-3,4-diols are combined as constituent units by condensation and polymerization. In addition, the proanthocyanidins of the present invention include high molecular weight proanthocyanidins such as proanthocyanidins, prodelphinidins, and propelargonidins composed of trimers, tetramers, and 10 to 30-mers of structural units, and their steric forms. Isomers and the like are also included. In addition, this proanthocyanidins are classified into several types based on the difference in binding mode. One is a proanthocyanidin B type in which two or more flavan-3-ol structures are bonded via the 4β position → 8 position or the 4β position → 6 position, and the other is a flavan-3-ol structure. It is a proanthocyanidin A type in which the bonds of 4β position → 8 position and 2β position → O → 7 position or 4β position → 6 position and 2β position → O → 7 position exist in at least one position. There are both proanthocyanidin A type and proanthocyanidin B type in cranberries.

The proanthocyanidin A type in the present invention has flavan-3-ol or flavan-3,4-diol as a structural unit, and the forms in which these are condensed or polymerized are 4β position → 8 position, and 2β position → O → 7 position, Alternatively, it is a compound group in which the bonds of 4β position → 6 position and 2β position → O → 7 position exist in at least one position, and are substances represented by the following structural formulas (1) and (2).
Structural formula (1):

構造式（２）：

Structural formula (2):

［式中、Ｒ₁は水素原子又は水酸基であり、Ｒ_２、Ｒ_３、Ｒ_４は水素原子又はガロイル基である。］

[Wherein, R ₁ is a hydrogen atom or a hydroxyl group, and R ₂ , R ₃ and R ₄ are a hydrogen atom or a galloyl group. ]

The proanthocyanidin A type ₂ described in the present invention refers to a dimer (epicatechin- (4β-8,2β-O7) -epicatechin) in which epicatechin is polymerized in the 4β position → 8 position and 2β position → O → 7 position. (Structural formula (3)).
Structural formula (3):

The proanthocyanidin B type ₂ described in the present invention refers to a dimer (epicatechin- (4β-8) -epicatechin) in which epicatechin is polymerized from the 4β-position to the 8-position (structural formula (4)).
Structural formula (4):

(Cranberry peel / cranberry pomez)
A cranberry is a small fruit tree that originates in both Europe and North America. In the present invention, its original species and cultivars can be used. Cultivation varieties are native to the state of Winsconsin, USA, and are fruit trees of the height of about 20 cm of Vaccinium macrocarpon (Vaccinium macrocarpon) or Occococcus (Vaccinium oxycoccus). Stevens, Benlear, Pilgrim, Early black, Bergman and the like.

  The cranberry extract of the present invention can be obtained by hot water extraction from cranberry peel or cranberry pomez. Cranberry peel refers to the outer part that covers cranberry fruit. Cranberry pomez is a residue after squeezing cranberry concentrated juice, and the cranberry peel occupies most of the pomez. In order to increase the extraction efficiency, it is also possible to pulverize the cranberry peel or cranberry pomez beforehand. An extract containing a large amount of proanthocyanidins A can be efficiently obtained through a hot water extraction process from cranberry peel or cranberry pomez, but considering the cost, it is cheap and easily available. Pomez is preferably used.

(Hot water extraction)
The cranberry extract which is the present invention can be obtained by extracting the cranberry peel or cranberry pomez with hot water. The extraction conditions vary depending on the production area of the cranberries and the harvest time, but can be appropriately selected within the following conditions.

  The solvent to be extracted is hot water. The temperature of hot water is preferably in the range of 80 to 100 ° C, more preferably in the range of 90 to 100 ° C. When the temperature of the hot water is lower than 80 ° C., the extraction rate of proanthocyanidins is deteriorated.

  The extraction time is in the range of 2 to 5 hours, more preferably 3 to 4 hours. When the extraction time is up to 4 hours, the extraction rate of proanthocyanidins increases in proportion to the increase in the extraction time. However, when the extraction time exceeds 4 hours, the increase in the extraction rate gradually slows down and the extraction time exceeds 5 hours. Is also meaningless.

  The amount of the solvent is preferably in the range of 3 to 5 parts by weight with respect to 1 part by weight of cranberry peel or cranberry pomez. When the amount is less than 3 parts by weight, the extraction rate of proanthocyanidins is poor, and when the amount exceeds 5 parts by weight, the extraction rate of proanthocyanidins is high, but the extraction amount of monosaccharides is reduced, and astringency and astringency are increased. Therefore, it is not suitable for food.

  The solution after extraction with hot water obtained under the above conditions is filtered with a cloth, stainless steel filter, filter paper, celite or the like to remove cranberry pomez impurities, whereby the cranberry extract of the present invention can be obtained. Further, the cranberry extract after filtration may be concentrated, or may be pulverized by a treatment such as spray drying treatment or freeze drying treatment. By pulverizing, it becomes easy to add to food and it is easy to grasp the amount added, so it is excellent in convenience. When powdering, the filtered hot water extract is concentrated, 20 to 100% (w / w) excipient is added to the solid content of the extract, and spray-dried after sterilization However, it is preferable because it is easy and economical.

(Cranberry extract)
By the above process, a cranberry extract containing 0.6 parts by weight or more of proanthocyanidin A ₂ type and 1% or more of monosaccharide (w / w) with respect to 1 part by weight of proanthocyanidin B ₂ type is obtained. Can do. Since the said cranberry extract is excellent in flavor compared with the extract containing many conventional proanthocyanidins A type ₂ , it can be easily used as a raw material for foods and beverages. In addition, since it contains proanthocyanidin A type ₂ above a certain concentration, it is applied to health foods or pharmaceuticals that have an inhibitory effect on the activity of hyaluronidase, anti-allergic or anti-inflammatory action, and prevent bacteria from causing urinary tract infections. Can be expected.

  Examples of the present invention will be described below, but the present invention is not limited thereto.

(Examination time)
The extraction time was examined. 3 parts by weight of water (150 ml) is added to 1 part by weight (50 g; 36 g of water) of cranberry pomez, which is the residue of American cranberry juice, heated to 90 ° C., and heated to reflux for 1 to 6 hours under reflux cooling. went. The extraction solution obtained after hot water extraction was filtered using Celite (Standard Supercell; manufactured by Wako Pure Chemical Industries, Ltd.), and then concentrated under reduced pressure until the volume became less than 50 ml to obtain a concentrated solution of the extraction solution. Next, water was added to the concentrated solution so that the volume became 50 ml, and the solid content contained in the concentrated solution was determined.

  The solid content was calculated by the following method. After appropriately diluting with water so that the concentration of the solid content contained in the concentrate was 20 mg / ml or less, 5 ml of the diluted solution was placed in a beaker and allowed to stand at 110 ° C. for 3 hours. Then, it dried in the desiccator for 1 hour or more at room temperature, the weight of the beaker after drying was calculated | required, the weight of the tare was subtracted from the weight, and solid content concentration of the concentrate was calculated | required.

To the concentrated solution, starch (Stabilose S-10; manufactured by Matsutani Chemical Co., Ltd.) having an amount equal to the solid content determined by the above method was added as an excipient, and freeze-dried to obtain a powdered cranberry extract. Subsequently, the total amount of proanthocyanidins (w / w), the proanthocyanidin ratio (A ₂ / B ₂ ), and the amount of monosaccharides (w / w) contained in the cranberry extract were determined.

  The total amount of proanthocyanidins was calculated from the total amount of flavanols determined by the hydrochloric acid-vanillin method (RB Broadhurst, et al, J. Ssi. Food Agric., Vol. 29, 788-794 (1978)). The subtracted value was used.

  The proanthocyanidin ratio was used by attaching a fluorescence detector (manufactured by Hitachi High-Technologies Corporation) to HPLC (manufactured by JASCO Corporation). The measurement was performed under the following conditions.

Eluent A; 10% acetonitrile solution containing 0.1% trifluoroacetic acid Eluent B; 40% acetonitrile solution containing 0.1% trifluoroacetic acid Eluent flow rate; 1.0 ml / min column; ODS column Column oven Temperature; 40 ° C
Excitation wavelength: 276 nm
Fluorescence wavelength; 316nm
Injection volume: 10 μL
Concentration gradient condition; Eluent B was increased linearly from 0% to 100% from the start of analysis to 40 minutes. Thereafter, the eluent B was kept at 100% until 50 minutes.

  The amount of monosaccharide was measured by HPLC using the phosphate-phenylhydrazine method (Hisao Inoue et al., 17, 7-17 (2003)), and indicated by the sum of arabinose, fructose, glucose, galactose, and sucrose. . The measurement by HPLC was performed under the following conditions.

Eluent A; 95% acetonitrile containing 0.5% phosphoric acid Eluent B; 75% acetonitrile containing 0.5% phosphoric acid Eluent flow rate; 1.0 ml / min Column oven temperature;
Excitation wavelength: 330 nm
Fluorescence wavelength: 470nm
Injection volume: 5μL
Concentration gradient condition: Eluent B 0% was kept for 20 minutes from the start of analysis. From 20 minutes to 40 minutes, the eluent B was linearly increased to 60%. The eluent B was held at 60% for 40 to 50 minutes. Thereafter, the eluent B was increased linearly from 60% to 100% in 55 minutes. Thereafter, the eluent B was kept at 100% until 65 minutes.

  Table 1 shows the total amount of proanthocyanidins, the ratio of proanthocyanidins, and the amount of monosaccharides at each extraction time.

As shown in Table 1, when hot water extraction was performed at 90 ° C., the amount of proanthocyanidins extracted increased with an increase in the extraction time when the extraction time was 4 hours or less. Change is no longer seen. Also, the extraction time exceeds 4 hours, A ₂ / B ₂ ratio also because it is 0.6 or less, proanthocyanidin A ₂ type extraction efficiency was confirmed to be reduced. For the amount of monosaccharides, no effect due to changes in extraction time was observed.

  From the above results, it can be seen that the optimum extraction time is 2-5 hours, most preferably in the range of 3-4 hours.

(Examination of amount of extraction solvent)
The amount of extraction solvent was examined. In accordance with the method of Example 1, 2 to 8 parts by weight (100 to 400 ml) of water was added to 1 part by weight (50 g; 36 g of water) of cranberry pomez, heated to 90 ° C., and refluxed for 2 hours. And hot water extraction was performed. Thereafter, lyophilization was carried out in the same manner as described in Example 1 to obtain a cranberry extract. The total amount of proanthocyanidins contained in the extract (w / w) and the ratio of proanthocyanidins (A ₂ / B ₂ ) And the amount of monosaccharide (w / w) was determined. The results are shown in Table 2.

  As shown in Table 2, when the extraction solvent is less than 3 parts by weight relative to 1 part by weight of cranberry pomez, the proanthocyanidin content after extraction is small, and when the extraction solvent exceeds 5 parts by weight, the monosaccharide content after extraction is The astringent and astringent taste of the resulting cranberry extract is reduced, making it unsuitable for food use.

  From the above results, it can be seen that the optimum amount of solvent is in the range of 3 to 5 parts by weight per 1 part by weight of cranberry peel or cranberry pomez.

(Examination of extraction temperature)
The extraction temperature was examined. According to the method of Example 1, 3 parts by weight (150 ml) of water was added to 1 part by weight (50 g; 36 g of water) of American cranberry pome and heated at 60 ° C., 70 ° C., 80 ° C., or 90 ° C. Hot water extraction was performed under reflux cooling for 4 hours. Thereafter, lyophilization was carried out in the same manner as described in Example 1 to obtain a cranberry extract. The total amount of proanthocyanidins contained in the extract (w / w) and the ratio of proanthocyanidins (A ₂ / B ₂ ) And the amount of monosaccharide (w / w) was determined. The results are shown in Table 3.

  As shown in Table 3, it was confirmed that when the extraction temperature was lowered, the extraction efficiency of proanthocyanidins was lowered. On the other hand, no difference in the ratio of proanthocyanidins and the amount of monosaccharides due to temperature changes was confirmed.

  From the above results, it is understood that the optimum extraction temperature is in the range of 80 to 90 ° C.

(Examination of differences depending on the production area of cranberries)
A comparative study was conducted to determine whether the cranberry extract differs depending on the production area of the cranberries. According to the method of Example 1, 3 parts by weight (150 ml) of water was added to 1 part by weight (50 g; 36 g of water) of cranberry pomez, which is a residue of chili cranberry juice, and 60 ° C, 70 ° C, 80 ° C, Or it heated at 90 degreeC and hot water extraction was performed under reflux cooling for 4 hours. Thereafter, lyophilization was carried out in the same manner as described in Example 1 to obtain a cranberry extract. The total amount of proanthocyanidins contained in the extract (w / w) and the ratio of proanthocyanidins (A ₂ / B ₂ ) And the amount of monosaccharide (w / w) was determined. The results are shown in Table 4.

As shown in Table 4, it was confirmed that a cranberry extract rich in proanthocyanidins A type ₂ and monosaccharides can be obtained in the range of 80 to 90 ° C., regardless of the production area of cranberries. .

(Examination of extraction solvent)
The extraction solvent was examined. According to the method of Example 1, 3 parts by weight (150 ml) of water was added to 1 part by weight (50 g; 36 g of water) of cranberry pomez, which is a residue of squeezed American cranberries, and heated at 90 ° C. for 3 hours. Hot water extraction was performed under reflux cooling, and lyophilization was performed in the same manner as in the method described in Example 1 to obtain a cranberry extract (Invention 1) according to the present invention. On the other hand, by using 30%, 50%, or 90% ethanol as a solvent and extracting in the same manner as described above, comparative examples (comparative examples 1 to 3) were obtained. The total amount of proanthocyanidins (w / w), the proanthocyanidin ratio (A ₂ / B ₂ ), and the amount of monosaccharides (w / w) contained in each extract were measured in the same manner as in Example 1. The results are shown in Table 5.

As shown in Table 5, it was confirmed that the cranberry extract of the present invention contained abundant monosaccharides, although the concentration of proanthocyanidins and the extraction efficiency of proanthocyanidin A type ₂ were lower than those of the comparative examples. It was done.

  Subsequently, the sensory evaluation test of each extract was performed. The sensory evaluation test was performed by a blind test for three panelists, and the taste and the overall taste were evaluated. The results are shown in Table 6.

  As shown in Table 6, it was confirmed that the cranberry extract according to the present invention is less astringent than the comparative example and exhibits a good flavor with an excellent balance between sourness and sweetness. On the other hand, it was confirmed that the extract of the comparative example was very astringent and unsuitable for use in foods. This is because extraction of cranberry pomes using ethanol extracts a large amount of flavanols such as quercetin contained in cranberries due to the polarity of the solvent, and extraction of monosaccharides that is efficiently extracted when extracted using hot water It is thought that the flavor was impaired by the worsening of the taste.

Claims (2)

Containing proanthocyanidin 4% (w / w) or more, and, with respect to the proanthocyanidin B ₂ type 1 part by weight contained in the proanthocyanidin state, and are proanthocyanidin A _2-inch 0.6 parts by weight or more, A cranberry extract containing 10% (w / w) or more of a monosaccharide . Cranberry pericarp or to cranberry port female 1 part by weight, after heating with the addition of water 3 to 5 parts by weight, in the range of 80-90 ° C., cranberry of claim 1, wherein the extracting 2-5 hours A method for producing an extract.