JP2000063827A - Manufacture of antioxidant - Google Patents

Abstract

PROBLEM TO BE SOLVED: To increase a content rate of neochlorogenic acid and antioxidation activity by treating an aqueous solution containing chlorogenic acid with an alkali. SOLUTION: A raw material of chlorogenic acid is preferably coffee or sunflower because they are easily available. Crushed seeds of sunflower are extracted with about 5-10 times by weight of water or an alcohol containing water at room temperatures or elevated temperatures. The substance extracted with water is directly treated by use of sodium hydroxide, potassium hydroxide or the like, at room temperatures under pH of 10-12, while the substance extracted with the alcohol is treated in the same way after the content of alcohol is reduced to about not more than 1% by weight by means of distillation or the like. After 3 minutes to 1 hour pH is adjusted to 3-4 by adding a mineral acid such as hydrochloric acid and sulfuric acid or an organic acid such as acetic acid and then condensation is conducted under reduced pressure to produce an antioxidant after isomerization is finished. The ratio of the chlorogenic acid to isochlorogenic acid to neochlorogenic acid is changed from 3-4:1:1 to 1:1:1 by alkali treatment to give highly active antioxidation.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a neochlorogenic acid content ratio by subjecting an aqueous solution containing chlorogenic acids, particularly a water extract or a hydroalcoholic extract of a plant containing chlorogenic acids such as sunflower and coffee to alkali treatment. It relates to a method for producing an enhanced antioxidant.

[0002]

BACKGROUND ART Chlorogenic acids are used for coffee beans, fruits,
It is known as a natural antioxidant that is widely distributed in the plant kingdom such as vegetables, and is mainly used as an antioxidant in the food field. For example, chlorogenic acids are used to prevent discoloration such as fading and browning of foods and drinks (Japanese Patent Publication No. 1-22872, Japanese Patent Laid-Open No. 5-28772).
-32909, JP-A-6-93199), prevention of flavor deterioration (JP-A-6-38723), stabilization of vitamin C (JP-A-6-9603), and some other examples disclosed. Has been done.

Examples of plants containing chlorogenic acids include sunflower seeds, coffee beans, cowberry leaves, mate tea, elderberry leaves, tobacco leaves, unripe apples, pears, and sweet potatoes. Chlorogenic acids in these plants. The content ratio of each component of is various. The compounds collectively referred to as chlorogenic acids include, in addition to chlorogenic acid, isochlorogenic acid and neochlorogenic acid, which are positional isomers of chlorogenic acid.

The present inventors have previously studied the relative antioxidant activity of each component of chlorogenic acids, and found that among the chlorogenic acids, neochlorogenic acid exhibits the strongest antioxidant activity, and the content of neochlorogenic acid was found. It is preferable to use chlorogenic acids having a high ratio.
It has already been reported in Japanese Patent Publication No. 43465.

However, the content of neochlorogenic acid in the water extract of a plant containing chlorogenic acids or the hydroalcoholic extract varies depending on each plant, the production area and the harvesting time, and the content or ratio of neochlorogenic acid is artificially changed. I couldn't control it.

[0006]

The problem to be solved by the present invention is to artificially treat an aqueous solution of a chlorogenic acid derived from a natural product so as to have a stronger antioxidant activity than other chlorogenic acid isomers. It is intended to provide a method for producing an antioxidant having a high chlorogenic acid content and high antioxidative activity, and an antioxidant having an increased neochlorogenic acid content by the production method.

[0007]

Means for Solving the Problems As a result of intensive studies, the present inventors have found that by treating chlorogenic acid with an alkali, it can be easily converted into neochlorogenic acid having a high antioxidant activity, and a plant water extract or It was found that by treating the hydrous alcoholic extract with an alkali, neochlorogenic acid can be increased without decreasing the total amount of chlorogenic acids, and an antioxidant with high antioxidant activity can be obtained, and the present invention has been completed. It was

That is, the present invention provides a method for producing an antioxidant characterized by increasing the content ratio of neochlorogenic acid by subjecting (a) an aqueous solution containing chlorogenic acids to an alkali treatment, and (b) chlorogenic acids. Aqueous solution containing, characterized in that it is a water extract or hydrous alcohol extract solution of a plant containing chlorogenic acids, (a) a method for producing an antioxidant,

(C) A method for producing an antioxidant according to (a) or (b), characterized in that the alkali treatment is carried out at a pH of 10 to 12, and (d) after the alkali treatment, the treatment liquid is neutralized. A method for producing an antioxidant according to (a) or (b), characterized in that the pH of the treatment liquid is treated to 3 to 4.

(E) An aqueous extract of a plant containing chlorogenic acids is treated with an alkali at a pH of 10 to 12, and then alcohol is added to remove the resulting precipitate.
(A) or the method for producing an antioxidant according to (b),

(F) The plant containing chlorogenic acids is sunflower, and the method for producing an antioxidant according to (ii) or (ii),

(H) The method for producing an antioxidant according to (ii) or (ii), characterized in that the plant containing chlorogenic acids is coffee.

(B) An antioxidant containing chlorogenic acids in which the content ratio of neochlorogenic acid is increased by the production method described in any one of (a) to (h) above.

[0014]

BEST MODE FOR CARRYING OUT THE INVENTION The chlorogenic acids referred to in the present invention collectively refer to chlorogenic acid, isochlorogenic acid and neochlorogenic acid represented by the following chemical formulas. Chlorogenic acid and caffeic acid are available as commercial products, but isochlorogenic acid and neochlorogenic acid are not available as commercial products, and are extracted from plants such as sunflower seeds and then purified using a column chromatography method or the like. Obtained.

[0015]

[Chemical 1]

In the present invention, examples of raw materials for obtaining chlorogenic acids include sunflower seeds, coffee beans,
Examples include immature apples, tobacco leaves, pears, cowberry leaves, and sweet potatoes. The content of chlorogenic acids in these plants is considered to vary depending on the time of harvest, variety, and cultivated place, but one example is as shown in the table below.

[0017]

Of these, coffee beans, sunflowers belonging to the family Asteraceae and the genus Sunflower (Japanese name: Hyuga, Han name: Sunflower, Scientific name: Helianthus ann) are preferred because of their easy availability.
uusL.) so called fruit. It is mainly used as a raw material for sunflower oil and as a pet feed. Further, the meal after pressing the oil is usually used as a feed for livestock, which is also a suitable raw material of the present invention. In the present invention, it is more preferable that the lees after pressing the oil have the fat and oil components removed with hexane or another solvent. The outline of the method for obtaining chlorogenic acids from sunflower seeds is as follows.

First, sunflower seeds are crushed by a crusher or the like. It may be used as it is if the shell is already crushed by crushed oil cake or the like. Next, for these, about 5
Extract by adding 10 times (weight) water or hydrous alcohol. When extracting with water, it may be at room temperature, but 50 to 10
It is more preferable to use hot water heated to 0 ° C. or an autoclave (for example, 120 ° C. × 2 atm).

When hydrous alcohol is used, for example, an alcohol solution of 80% or less, preferably 65% or less, is used at room temperature to 70 ° C., preferably 5%.
Extract by heating at around 0 ° C. The extraction time is preferably 1 to 2 hours or more. Usually, this extract is concentrated under reduced pressure and then dried to be powdered or made into a paste to finish. As the extraction alcohol, for example, methyl alcohol, ethyl alcohol, propyl alcohol, isopropyl alcohol or the like can be used.

The extract containing chlorogenic acids is used as it is in the case of water extraction, and in the case of hydrous alcohol extract, after the content of the alcohol is adjusted to, for example, about 1% by weight or less by means such as distillation. Used for alkaline treatment. By performing the alkali treatment of the present invention, chlorogenic acid is isomerized to isochlorogenic acid or neochlorogenic acid, and the three isomers are in an equilibrium state.

Examples of the alkaline substance used for the alkali treatment include sodium hydroxide, potassium hydroxide,
Examples thereof include calcium hydroxide, sodium carbonate, sodium hydrogen carbonate, potassium carbonate and the like. At the time of addition, sodium hydroxide and potassium hydroxide can be preferably mentioned rather than carbonate-based alkalis that generate carbon dioxide gas. Examples of the aqueous alkaline solution used in the present invention include aqueous solutions of about 1 to about 20% by weight of the above alkaline substances.

When the alkali treatment is carried out, the pH of the extract is usually 3 to 4, and the pH is adjusted using the above alkaline aqueous solution. However, if the pH is less than 9, isomerization hardly proceeds, When it is more than 12, a hydrolysis reaction occurs, is sparingly soluble in water, and a large amount of caffeic acid having a lower antioxidative activity than that of chlorogenic acid, isochlorogenic acid and neochlorogenic acid is produced. Adjust pH to 10-12. Particularly preferably, the pH is 11 to
Adjust to 12.

The reaction temperature for carrying out the alkali treatment is usually room temperature (about 20 ° C.), but at a high temperature of 80 ° C. or higher, a hydrolysis reaction occurs and caffeic acid is generated.
Room temperature to 50 ° C is preferable.

Further, if the alkali treatment time is 1 minute or less, the reaction does not proceed sufficiently, and if it is 3 hours or more, a hydrolysis reaction and decomposition of the active ingredient occur, so that the reaction time is 3 minutes to 1 minute.
Time is preferable.

The alkaline treatment liquid obtained as described above is usually adjusted to acidic pH with an acidic aqueous solution to terminate the isomerization reaction with alkali. At the end of the isomerization reaction, if the pH is higher than 5, the solution will change color and p
When H is less than 2, a hydrolysis reaction with an acid occurs and caffeic acid is produced, so that the adjusted pH is 3 to
4 is preferable.

As the acidic aqueous solution used in the present invention, an approximately 1 to 20% by weight aqueous solution of an acidic substance can be exemplified. Further, as the above-mentioned acidic substance, for example, hydrochloric acid, sulfuric acid, nitric acid, mineral acids such as phosphoric acid, acetic acid, citric acid,
Organic acids such as malic acid and ascorbic acid can be mentioned, and hydrochloric acid and sulfuric acid are preferable.

The antioxidant of the present invention can be obtained by concentrating under reduced pressure the above alkaline treatment liquid whose pH is adjusted to 3 to 4 with an acidic aqueous solution. If necessary, the alkaline treatment liquid having a pH adjusted to 3 to 4 may be desalted and then concentrated under reduced pressure to obtain a composition with an appropriate diluent or carrier.

Examples of such diluents or carriers include solid diluents or carriers such as dextrin, glucose, cyclodextrin and sucrose, water,
Liquid diluents or carriers such as ethanol, propylene glycol, glycerin and surfactants can be mentioned. That is, the antioxidant of the present invention can be processed into a liquid form, a powder form, a granule form or any other suitable dosage form by using such a diluent or carrier.

The antioxidant of the present invention can be added to, for example, foods and drinks, fragrances, pigments, cosmetics, pharmaceuticals, etc.
Oxidation of these materials can be prevented. The amount of the antioxidant of the present invention to be added to foods and drinks, flavors and dyes is not particularly limited, and can be widely selected depending on the kind of foods and drinks and flavors, dyes, etc. It is suitable to add within the range of about 0.01 to about 1.0% by weight of food and drink.

The isomer ratio of chlorogenic acids in plants containing chlorogenic acids has not been reported in many cases. For example, the isomer content ratio of chlorogenic acid: isochlorogenic acid: neochlorogenic acid in sunflower seed extract is Usually, it is 3 to 4: 1: 1, and the isomer content ratio in the coffee bean extract is usually 10: 1: 1 (measured value is 1: 1).
0: 3: 4), and the content of neochlorogenic acid is low in both cases.

According to the present invention, it is possible to artificially bring the isomer ratio of chlorogenic acids to a value close to 1: 1: 1 by increasing the content of neochlorogenic acid, which has the highest antioxidant activity. A highly active antioxidant can be produced.

[0033]

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

Example 1 0.3 g of chlorogenic acid (Tokyo Kasei Special Grade, manufactured by Tokyo Chemical Industry Co., Ltd.) was dissolved in 100 ml (pH 2.8), and the pH was adjusted to 11 with a 15% sodium hydroxide aqueous solution. And stirred at room temperature for 3 minutes. Two
The pH was adjusted to 2.8 with a 0% aqueous hydrochloric acid solution, and the mixture was concentrated under reduced pressure with a rotary evaporator.

As a result, 0.31 g of the product (a) was obtained. Table 1 shows the changes in chlorogenic acids due to the alkali treatment. The content in the table indicates the weight / volume% of chlorogenic acids in the solid content, and the ratio indicates the content of each component when the chlorogenic acid content is 1. The high performance liquid chromatography analysis method is as follows.

(High-performance liquid chromatography analysis method) Column used: Inertsil (registered trademark of GL Sciences Inc.) ODS-2, 5 μ 4.6 φ × 250 mm Solvent: water-methanol = 70-30 (volume ratio) pH 2.7 (phosphorus) Adjusted with acid) Flow rate: 0.7 ml / min

In the table, each component of chlorogenic acids is abbreviated as follows. Chlorogenic acid: Chloro., Neochlorogenic acid: Neo., Isochlorogenic acid: Iso., Caffeic acid: Caff.

[0038]

[Table 1]

Reference Example 1 1000 ml of distilled water was added to 100 g of sunflower seeds, and chlorogenic acids were extracted at 90 to 95 ° C. for 3 hours. After filtration with a Buchner funnel, the mixture was concentrated under reduced pressure with a rotary evaporator. To this concentrated dried product, 84 ml of 70% aqueous ethanol solution was added and stirred for 1 hour. After filtration, the filtrate was concentrated under reduced pressure with a rotary evaporator. As a result, 7.6 g of sunflower seed extract was obtained.

Example 2 5 g of the sunflower seed extract obtained in Reference Example 1 was dissolved in 100 ml of distilled water (pH
3.8), the pH was adjusted to 11 with a 15% aqueous sodium hydroxide solution, and the mixture was stirred at room temperature for 3 minutes. The pH was adjusted to 3 to 4 with 20% hydrochloric acid, and concentrated under reduced pressure with a rotary evaporator. As a result, 5.2 g of product (b) was obtained. Table 2 shows the analytical values of the obtained chlorogenic acids.
The content in the table indicates the weight / volume% of chlorogenic acids in the solid content, and the ratio indicates the content of each component when the chlorogenic acid content is 1.

[0041]

[Table 2]

Example 3 1000 ml of distilled water was added to 100 g of sunflower seeds, and chlorogenic acids were extracted at 90 to 95 ° C. for 3 hours. After filtration with a Buchner funnel, 15
The pH was adjusted to 11 with an aqueous sodium hydroxide solution, and the mixture was stirred at room temperature for 3 minutes. P of this filtrate with 20% hydrochloric acid
Set H to 3-4, and use Bri on a rotary evaporator.
It concentrated under reduced pressure until x became 20. 99.
58 ml of 5% ethanol was added and stirred for 1 hour to precipitate insoluble matter. After filtering the precipitate, it was concentrated under reduced pressure with a rotary evaporator to obtain 7.8 g of the product (c).

Example 4 To 100 g of coffee beans was added 1000 ml of distilled water, and chlorogenic acids were extracted at 85 to 90 ° C. for 3 hours. After filtration with a Buchner funnel, the mixture was concentrated under reduced pressure with a rotary evaporator. 140 ml of 70% aqueous ethanol solution was added to this concentrated dry solid, and the mixture was stirred for 1 hour. After filtration, the filtrate was concentrated under reduced pressure with a rotary evaporator to obtain 15.8 g of a coffee bean extract.

5 g of this coffee bean extract was dissolved in 100 ml of distilled water (pH 5.5), the pH was adjusted to 11 with a 15% aqueous sodium hydroxide solution, and the mixture was stirred at room temperature for 3 minutes. The pH was adjusted to 5.5 with 20% hydrochloric acid, and concentrated under reduced pressure with a rotary evaporator. As a result,
1 g of product (d) was obtained. Table 3 shows the analytical values of the obtained chlorogenic acids. The content in the table indicates the weight / volume% of chlorogenic acids in the solid content, and the ratio indicates the content of each component when the chlorogenic acid content is 1.

[0045]

[Table 3]

Example 5 10 parts by weight of the product (b) obtained in Example 2 and dextrin (NSD # 500, D)
19 parts by weight of E10-13, manufactured by Nippon Sanyo Kogyo Co., Ltd.) were mixed to obtain an antioxidant composition (a).

(Example 6) 20 parts by weight of the product (b) obtained in Example 2, 30 parts by weight of ethanol (special grade reagent, manufactured by Wako Pure Chemical Industries, Ltd.), and 50 parts by weight of water were mixed to give an anti-oxidant agent. An oxidant composition (b) was obtained.

Example 7 10 parts by weight of the product (d) obtained in Example 4 and dextrin (NSD # 500, D)
79 parts by weight of E10-13, manufactured by Nippon Sanyo Kogyo Co., Ltd.) were mixed to obtain an antioxidant composition (c).

(Test Example of Antioxidant Activity) The products (a) to (d) of Examples 1 to 4 were added to 0.01%, 0.115%,
0.115%, 0.037% added, and Examples 5-7
0.33% of each of the antioxidant compositions (a) to (c) are added, and the results of the test by Test method 1 (Rhodan iron activity test method) are shown in Table 7, and Test method 2 (of β-carotene). Table 8 shows the results of the test conducted by the anti-fading test method). BHA in Table 7 is an abbreviation for butylhydroxyanisole, and storage conditions for the test in Table 8 are pH 6, 3500 l.
ux, 20 ° C.

(Test method 1) Rhodan iron activity test method Autoxidation of linoleic acid was carried out, and the peroxide value was measured by the rhodan iron method (thiocyanate method). That is, 2.5%
1 ml of linoleic acid (99.5% ethanol solution),
Put a mixed solution of 2 ml of 0.05M phosphate buffer (pH 7.0) and 2 ml of distilled water into a screw cap test tube (φ34 × 80 mm), add the test sample to the target concentration, and shade at 37 ° C. Saved below.

Samples stored over time 0.1 ml, 75%
Exactly 3 minutes after adding 0.05 ml of 0.02 M ferrous (II) chloride (3.5% hydrochloric acid aqueous solution) to a mixed solution of 4.8 ml of aqueous ethanol solution and 0.05 ml of ammonium thiocyanate solution, 500 nm The absorbance at was measured. In this method, the absorbance of 0.3 is usually regarded as the rancid branch point of linoleic acid, and therefore the antioxidant activity was evaluated using the period (days) until the peroxide value of the sample reached this value as the number of induction days.

(Test Method 2) Test Method for Preventing Fading of β-Carotene β-Carotene is widely used as an edible pigment, but it is oxidatively decomposed and fades under conditions such as light, heat, oxygen and humidity. The anti-fading effect of β-carotene was examined as an antioxidant test for chlorogenic acids. That is, after mixing 0.03% by weight of water-soluble β-carotene (manufactured by Sankyo Co., Ltd.) and 0.1% by weight of potassium sorbate (preservative), pH was 20 ° C and pH was 20 ° C.
6, stored under 3500 lux, and the absorbance at 500 nm was measured over time. The absorbance at 500 nm was used as the amount of β-carotene, and the residual rate was used for the evaluation of the activity. β-
The carotene residual rate is calculated by the following formula.

[0053]

[Formula 1]

Example 8 Alkaline Treatment pH Conditions The sunflower seed extract obtained in Reference Example 1 was treated with an alkali treatment time: 3 minutes, an alkali treatment temperature: 20 ° C., a treatment solution concentration: 5%, 15%. PH 8 ~ with sodium hydroxide solution
Table 4 shows the results of processing in No. 13. From Table 4 pH 11
It can be seen that the alkaline treatment at ~ 12 maximizes the content of neochlorogenic acid.

[0055]

[Table 4]

(Example 9) Alkaline treatment time The sunflower seed extract obtained in Reference Example 1 was adjusted to pH 11 with a 15% aqueous sodium hydroxide solution, the alkaline treatment temperature was 20 ° C, and the concentration of the treated solution was 5%. The results of treatment for 7 minutes to 7 hours are shown in Table 5. It can be seen from Table 5 that the content of neochlorogenic acid is highest in the treatment time of 3 minutes to 1 hour.

[0057]

[Table 5]

(Example 10) Alkaline treatment temperature The sunflower seed extract obtained in Reference Example 1 was adjusted to pH 11 with a 15% aqueous sodium hydroxide solution, and the alkali treatment time was 3 minutes and the concentration of the treated solution was 5% to 20 to 20%. Table 6 shows the results of the treatment at 80 ° C. From Table 6, it was found that the treatment at 20 to 50 ° C. gave the highest content of neochlorogenic acid.

[0059]

[Table 6]

[0060]

[Table 7]

[0061]

[Table 8]

[0062]

INDUSTRIAL APPLICABILITY The present invention artificially treats an aqueous solution of chlorogenic acids derived from natural products to increase the content of neochlorogenic acid, which has a stronger antioxidant activity than other chlorogenic acid isomers. It is possible to provide a method for producing an antioxidant having an oxidative activity, and an antioxidant having an increased content of neochlorogenic acid by the production method.

Claims (8)

[Claims] 1. A method for producing an antioxidant, which comprises increasing the content ratio of neochlorogenic acid by treating an aqueous solution containing chlorogenic acids with an alkali. 2. The method for producing an antioxidant according to claim 1, wherein the aqueous solution containing chlorogenic acids is a water extract or hydrous alcohol extract solution of a plant containing chlorogenic acids. 3. The method for producing an antioxidant according to claim 1, wherein the alkali treatment is carried out at pH 10-12. 4. The method for producing an antioxidant according to claim 1, wherein after the alkali treatment, the treatment liquid is neutralized to adjust the pH of the treatment liquid to 3 to 4. 5. The anti-oxidant according to claim 1, wherein the aqueous extract of the plant containing chlorogenic acids is alkali-treated at pH 10 to 12 and then alcohol is added to remove the resulting precipitate. Oxidizing agent manufacturing method. 6. The method for producing an antioxidant according to claim 1, wherein the plant containing chlorogenic acids is sunflower. 7. The method for producing an antioxidant according to claim 1, wherein the plant containing chlorogenic acids is coffee. 8. An antioxidant containing chlorogenic acids in which the content ratio of neochlorogenic acid is increased by the production method according to any one of claims 1 to 7.