阅读说明：本技术 根皮素糖苷类的精炼方法 (Refining method of phloretin glycosides ) 是由 铛史晶 中岛翔平 池田由香 于 2018-08-15 设计创作，主要内容包括：本发明涉及根皮素糖苷类的精炼方法。本发明的目的在于提供一种通过进一步提高纯度而能够精炼根皮素糖苷类的新技术。一种根皮素糖苷类的精炼方法,包括：向合成吸附树脂柱供给包含根皮素糖苷类的多酚,并将根皮素糖苷类吸附于该柱；以及在将乙醇的比例为整个溶液的0.1％以上且低于45％(V/V)的水与乙醇的混合溶液作为流动相而流动之后,将乙醇的比例为整个溶液的45％以上且低于100％(V/V)的水与乙醇的混合溶液、或者乙醇作为流动相而流动,得到包含根皮素糖苷类的馏分。(The invention relates to a refining method of phloretin glycosides. The purpose of the present invention is to provide a novel technique that enables phloretin glycosides to be refined by further increasing the purity. A method for refining phloretin glycosides, comprising: supplying a polyphenol containing phloretin glycosides to a synthetic adsorption resin column, and adsorbing phloretin glycosides to the column; and flowing a mixed solution of water and ethanol having an ethanol ratio of 0.1% or more and less than 45% (V/V) of the total solution as a mobile phase, and then flowing a mixed solution of water and ethanol having an ethanol ratio of 45% or more and less than 100% (V/V) of the total solution or ethanol as a mobile phase to obtain a fraction containing phloretin glycosides.)

1. A method for refining phloretin glycosides, comprising:

supplying a polyphenol containing phloretin glycosides to a synthetic adsorption resin column, and adsorbing phloretin glycosides to the column; and

after a mixed solution of water and ethanol having an ethanol ratio of 0.1% or more and less than 45% (V/V) of the total solution is flowed as a mobile phase, a mixed solution of water and ethanol having an ethanol ratio of 45% or more and less than 100% (V/V) of the total solution or ethanol is flowed as a mobile phase, and a fraction containing phloretin glycosides is obtained.

2. The method for refining phlorizin glycosides according to claim 1, wherein,

the polyphenol containing phloretin glycosides is derived from apple.

3. The method for refining phlorizin glycosides according to claim 1 or 2, wherein,

the synthetic adsorption resin column is an aromatic resin column.

4. A beverage, wherein,

the content of phloretin glycosides is above 60mg/l, and the content of chlorogenic acid is below 120 mg/l.

5. A beverage, wherein,

phloretin glycosides are contained at 60mg/l or more, and proanthocyanidins are contained at 720mg/l or less.

6. A solid food, wherein,

the phloretin glycoside is contained at a concentration of 25g/kg or more, and chlorogenic acid is contained at a concentration of 50g/kg or less.

7. A solid food, wherein,

phloretin glycosides are included at a concentration of 25g/kg or more, and proanthocyanidins are included at a concentration of 300g/kg or less.

Technical Field

The invention relates to a refining method of phloretin glycoside.

Background

Hyperuricemia is a state in which the amount of uric acid present in blood is higher than usual. Particularly, the number of patients is increasing in developed countries, and has been increasing in japan over the past few decades. Hyperuricemia is not only a cause of gout, but also has been pointed out to be associated with cardiovascular diseases such as hypertension. Therefore, it is considered to be important to further approximate uric acid levels to normal in order to improve the state of cardiovascular diseases, in addition to preventing gout.

On the other hand, apple polyphenol, which is an extract derived from apple fruits, is known to have an effect of improving hyperuricemia by inhibiting the amount of uric acid in blood (for example, patent document 1).

The apple polyphenol comprises phloretin glycoside such as phlorizin, procyanidin (procyanidin), catechin (catechin), epicatechin (epicatechin), chlorogenic acid, quercetin (quercetin), etc.

Among them, as a method for refining phloretin glycosides, for example, the method of patent document 2 is known.

Prior art documents

Patent document

Patent document 1: japanese patent laid-open No. 2006-16927;

patent document 2: japanese patent application laid-open No. 2014-506567.

Disclosure of Invention

The purpose of the present invention is to provide a novel technique that enables phloretin glycosides to be refined by further increasing the purity.

Generally, phloretin glycosides derived from apples are refined by using a column or the like.

As a result of intensive studies, the present inventors have found that a fraction containing phloretin glycosides at a higher ratio can be obtained by performing separation under predetermined conditions in refining using this column, and have completed the present invention.

[1] A method for refining phloretin glycosides, comprising:

supplying polyphenol containing phloretin glycosides to the synthetic adsorption resin column, and adsorbing phloretin glycosides to the corresponding column; and

after a mixed solution of water and ethanol having an ethanol ratio of 0.1% or more and less than 45% (V/V) of the total solution is flowed as a mobile phase, a mixed solution of water and ethanol having an ethanol ratio of 45% or more and less than 100% (V/V) of the total solution or ethanol is flowed as a mobile phase, and a fraction containing phloretin glycosides is obtained.

[2] The method for refining phloretin glycosides according to [1], wherein the polyphenol containing phloretin glycosides is derived from apple.

[3] The method for refining phloretin glycosides according to [1] or [2], wherein the synthetic adsorbent resin column is an aromatic resin column.

[4] A beverage contains phloretin glycoside 60mg/l or more and chlorogenic acid 120mg/l or less.

[5] A beverage contains phloretin glycosides 60mg/l or more and proanthocyanidins 720mg/l or less.

[6] A solid food contains phloretin glycoside 25g/kg above and chlorogenic acid 50g/kg below.

[7] A solid food contains phloretin glycosides 25g/kg or more and proanthocyanidins 300g/kg or less.

The present invention can provide a novel technique for refining phloretin glycosides by further improving the purity.

Drawings

FIG. 1 is a graph relating to sensory evaluation of beverages of the examples;

fig. 2 is a graph relating to sensory evaluation of the solid compositions of the examples.

Detailed Description

Hereinafter, one embodiment of the present invention will be described in detail.

The embodiment relates to a method for refining phloretin glycosides, which comprises the following steps: supplying a polyphenol containing phloretin glycosides to a synthetic adsorption resin column, and adsorbing phloretin glycosides to the column; and flowing a mixed solution of water and ethanol (hereinafter, referred to as 0.1% or more and less than 45% ethanol) having an ethanol ratio of 0.1% or more and less than 45% (V/V) of the total solution as a mobile phase, and then flowing a mixed solution of water and ethanol having an ethanol ratio of 45% or more and less than 100% (V/V) of the total solution or ethanol (hereinafter, referred to as 45% or more and less than 100% ethanol) as a mobile phase to obtain a fraction containing phloretin glycosides.

Phloretin glycosides are also called dihydrochalcone glycosides, and are a generic term for glycosides having phloretin as an aglycone. Examples of the phloretin glycosides include phlorizin, phloretin xyloglucoside, phloretin (phloretin-2 '-glucoside), phloretin-4' -glucoside, phloretin-4-glucoside, phloretin-2 ', 4' -diglucoside, phloretin-2 ', 4-diglucoside, phloretin-4, 4' -diglucoside, phloretin-2 ', 4, 4' -triglucoside, phloretin-2 '-xyloglucoside, phloretin-4-xyloglucoside, phloretin-2', 4 '-diglucoside, phloretin-2', 4-diglucoside, and phloretin-4, 4' -diglucoside, phloretin-2 ', 4, 4' -triglucoside, etc.

Phlorizin and phloretin xyloglucoside can be quantified, for example, by the method described in example 1. Further, phloretin glycosides may be treated with an enzyme and quantified as phloretin.

In the present embodiment, the phloretin glycoside-containing polyphenol is derived from apple fruits, for example. The apple is a plant of the genus Malus of the family Rosaceae, and its species and the like are not particularly limited in this embodiment. Examples of the inorganic pigment include Jinqian, Wanglin, Fuji, Qianyin, Hongyu, Australian green apple, Qianqiu, BeiDou, Luo, Snake fruit (Delcious), Sansha (sansa), autumn reflection, sunshine, world I, Red color Ji (Shinano Sweet), Asahi, India, Guoguang, Red Star (starking), and Hokkaido No. 9 (HAC-9), and one or more of them may be used.

The fruit may be any of mature fruit and young fruit, and is not particularly limited.

In addition, the polyphenol containing an apple-derived phlorizin glycoside is exemplified below, but the present invention is not limited thereto, and a polyphenol containing a phlorizin glycoside derived from another substance may be used in the treatment according to the refining method of the present embodiment.

The phloretin glycosides according to the present embodiment can be obtained, for example, by supplying an extract derived from apple juice containing phloretin glycosides, such as apple juice, to a column and refining the extract.

In the present specification, fruit juice refers to a liquid component obtained when fruit is squeezed into juice. Juicing refers to an operation of extracting and separating liquid components from fruits. For example, fruit juice is obtained by crushing fruit into an appropriate size and squeezing the crushed material. In the present embodiment, whether or not the treatment such as concentration or dilution with water is performed on the fruit juice is not particularly limited, and can be appropriately set by a person skilled in the art according to the column or the like used, for example.

An example of the method for refining phloretin glycosides according to the present embodiment will be described below.

First, apple juice is supplied to a synthetic adsorbent resin column, and phloretin glycosides are adsorbed to the column.

Then, a mixed solution of water and ethanol was refined as a mobile phase to obtain phloretin glycosides. Specifically, phloretin glycosides refined in the following manner were obtained: the method comprises flowing 0.1% or more and less than 45% ethanol as a mobile phase (first elution treatment), then flowing 45% or more and 100% or less ethanol as a mobile phase (second elution treatment), and recovering a fraction obtained by the second elution treatment.

In the refining method of the present embodiment, a synthetic adsorbent resin column can be used. The synthetic adsorbent used for the column is an insoluble three-dimensionally crosslinked structure polymer having large pores, which adsorbs various organic substances by van der Waals (van der Waals) force.

Examples of the synthetic adsorption resin column include aromatic resin columns such as styrene-divinylbenzene and acrylic resins, acrylic resin columns, and acrylonitrile resin columns, and those skilled in the art can appropriately set the column. Among them, an aromatic resin column is preferably used, and a styrene-divinylbenzene aromatic resin column is more preferably used, from the viewpoint of further improving the purity of the phloretin glycoside. Examples of the styrene-divinylbenzene aromatic resin column include a macroporous adsorbent resin (SEPABEADS, registered trademark, the same applies hereinafter) SP70, a formaldehyde resin (DIAION, registered trademark, the same applies hereinafter) HP20 (manufactured by mitsubishi chemical corporation), and the like.

In the first elution treatment, 0.1% or more and less than 45%, preferably 5% or more and less than 45%, more preferably 15% or more and 40% or less, and further preferably 20% or more and 40% or less of ethanol is flowed as a mobile phase. In this case, the volume of the eluent can be set to, for example, 1 to 3 for space velocity SV (space velocity, CV/h) and 1 to 4 for CV (column volume (resin volume)).

The pH of the eluent is also not particularly limited, and may be, for example, 7 or less.

In the second elution treatment, 45% or more and 100% or less, preferably 50% or more and 80% or less, and more preferably 60% or more and 80% or less of ethanol is flowed as a mobile phase. In this case, the volume of the eluent can be set to, for example, SV 1 to 3 and CV 2 or more. The pH of the eluent is also not particularly limited, and may be, for example, 7 or less.

For example, concentrated apple juice (Brix about 60 to 70) is diluted 3 times and supplied to an SP70 column, thereby adsorbing phloretin glycosides to the column. Specifically, the juice was entirely infused with SV 1. Next, water was passed at SV 1 and CV 1.5 to wash the column. After confirming that Brix (Brix) is reduced to 0.2% or less at the column outlet, 37% ethanol was passed at SV 1 and CV 3 (first elution), and 70% ethanol was passed at SV 1 and CV 3 (second elution).

Further, although the object to be adsorbed to the column is described by taking apple juice as an example, the present invention is not limited to this, and may be, for example, a crude polyphenol product obtained from apple juice or the like. The crude polyphenol product can be obtained by a known method, for example, by adsorbing components contained in apple juice on a synthetic adsorbent resin column and then subjecting the column to an elution treatment using a mixed solution of water and ethanol as a mobile phase, wherein the ratio of ethanol is 5 to 40% (V/V) of the total solution.

The phloretin glycoside thus obtained may be used as it is or may be subjected to a step such as concentration and drying and then used in the next step.

The phloretin glycoside according to the present embodiment is not particularly limited, but can be used for the production of pharmaceuticals, Quasi Drugs (Quasi-Drugs), foods and beverages, and the like containing phloretin glycoside. For example, the phloretin glycoside according to the present embodiment can be used as an active ingredient, and can be mixed with an excipient, a binder, a stabilizer, a disintegrant, a lubricant, a flavoring agent, a suspending agent, a coating agent, and any other optional ingredients as appropriate to prepare a preparation according to a predetermined method. The dosage form includes tablets, pills, capsules, granules, powders, syrups and the like, and tablets are preferred from the viewpoint of easy ingestion.

Alternatively, although not particularly limited, when the food or drink is produced in the form of a food or drink, the food or drink may be a food or drink for special use such as a specific health food or a nutritional functional food. Specific examples of the foods and drinks include nutritional supplementary foods (nutritional supplements in the form of tablets, etc.), refreshing beverages, yogurt, jelly, candy, bread, cookies, crackers (cracker), pizza crust, butter cake, ice cream, gummy candy, cakes and breads such as chewing gum, liquid foods, and foods for patients.

The beverage may be, for example, a canned beverage. Alternatively, the composition may be an alcoholic beverage to which alcohol is added.

In addition, the present invention is not limited to foods for humans, and may be added to feeds for poultry, for example.

As described above, according to the present embodiment, a novel technique capable of refining phloretin glycosides by further improving the purity thereof can be provided.

In the present embodiment, phloretin glycoside is obtained by column-refining polyphenol derived from apple or the like, and then eluting phloretin glycoside adsorbed on a resin. Conventionally, phloretin glycoside has been refined by performing a primary elution treatment using about 40% ethanol, but the method of the present embodiment is produced by a method that effectively utilizes fractions that have been conventionally disposed of as waste. The phloretin glycoside obtained by the method is a high-purity phloretin glycoside fraction which cannot be obtained by the prior art. Since pharmacological effects such as lowering of uric acid level cannot be obtained at the phloretin glycoside concentration contained in ordinary apple juice, even if the polyphenol fraction-extracted material having an increased phloretin glycoside concentration is dissolved and adjusted to a high concentration, there is a problem in taste (bitter and astringent tastes in many cases) due to the influence of inclusions, and there is a problem in that browning and the like occur. On the other hand, a beverage having a sufficient pharmacological effect can be produced for the first time by using the high-purity phloretin glycoside according to the present embodiment.

Chlorogenic acid and procyanidins are known as substances that reduce the taste or cause browning in apple polyphenols.

In addition, in the present specification, chlorogenic acid means 3-caffeoylquinic acid.

Procyanidins are catechins which are n polymers of catechins (n >1, n being an integer) or a generic name thereof, and do not include n ═ 1. The catechins include, but are not limited to, catechin, epicatechin, gallocatechin, and epigallocatechin. The proanthocyanidin is not particularly limited as long as it is a catechin n-polymer or a mixture thereof. In addition, natural procyanidins may be used, or synthesized procyanidins may be used.

Chlorogenic acid can be quantified by the method described in example 1, and procyanidins can be quantified by the method described in Japanese patent application laid-open No. 2009-156813.

For example, according to the present embodiment, the following beverage can be constituted: the total content of phloretin glycosides, preferably phloretin and phloretin xyloglucan is 60mg/l or more, the content of chlorogenic acid is 120mg/l or less and/or the content of procyanidins is 720mg/l or less. That is, according to the refining method of the present embodiment, the content of chlorogenic acid is 120mg/l or less and/or the content of proanthocyanidins is 720mg/l or less, and the content of phlorizin glycosides is 60mg/kg or more, and therefore, it is possible to provide a beverage containing more phlorizin glycosides, which is improved in taste by suppressing bitterness and astringency, and also suppressed in browning.

In addition, since phloretin glycosides have not been sufficiently refined by one column treatment, many times of refining have been repeated to form a nutritional supplement in the form of a tablet effective for reducing uric acid concentration in blood. As a result, the nutritional supplement could not be actually produced from the viewpoint of cost and the like.

On the other hand, according to the present embodiment, since phloretin glycosides having higher purity can be obtained, a nutritional supplement having lower cost can be produced. That is, according to the present embodiment, it is possible to contribute to realizing a nutritional supplement containing phloretin glycosides capable of reducing uric acid concentration in blood.

For example, according to the present embodiment, a food (solid food) can be provided, which is a solid such as a tablet containing phloretin glycosides, preferably phlorizin and phloretin xyloglucan in a total amount of 25g/kg or more, a chlorogenic acid content of 50g/kg or less, and/or a procyanidin content of 300g/kg or less.

In addition, since bitterness and astringency are further suppressed to further improve taste and further suppress browning, the following solid foods are preferred: the total content of phloretin glycosides, preferably phloretin and phloretin xyloglucan, is above 25g/kg, the content of chlorogenic acid is below 20g/kg and/or the content of procyanidins is below 150 g/kg.