JP3775584B2 - Production method of phenol-containing extract from Shiozo red perilla - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a method for producing a phenol-containing extract from salted red perilla, and more particularly to a method for producing an extract or powder containing a high concentration of phenols contained in a salted red perilla leaf.
[0002]
[Prior art]
It is known that phenols contained in red perilla leaves have antiallergic action (Monthly Food Chemical, Vol. 7, p. 37-42, 2000).
Some of the inventors established a method for producing an extract containing a high content of phenols from leaves of Labiatae plants such as red perilla, and in particular, clarified that rosmarinic acid is an active ingredient for antiallergic action ( Japanese Patent Application No. 2001-25277 Patent, Japanese Patent Application No. 2001-100346, WO02 / 62365 JP). However, red perilla leaves are highly seasonal foods and are not produced throughout the year, so it was impossible to produce extracts from fresh red perilla leaves throughout the year.
By the way, it is conventionally known that red perilla can be preserved by salting. However, it is unclear whether salted red perilla has the same level of phenols as fresh leaves, and even if phenols remain, it contains salt, so it can be used for various foods and drinks. It is difficult to apply.
Moreover, a method for efficiently extracting phenols simultaneously with desalting from salted red perilla has not been found.
[0003]
[Problems to be solved by the invention]
When applying a desalting method that is generally performed conventionally, the residual rate of phenols is reduced, so that the extract obtained from salted red perilla is used for functional foods and beverages, phenols, In particular, the rosmarinic acid content was insufficient.
An object of the present invention is to provide a method for efficiently producing red perilla extract or powder containing a high concentration of phenols from salted red perilla leaves.
[0004]
[Means for Solving the Problems]
As a result of intensive studies to solve the above problems, the present inventors have found an efficient method for producing red perilla extract or its powder containing a high concentration of phenols contained in the leaves of salted red perilla, The invention has been completed.
[0005]
The present invention according to claim 1 is characterized in that salted red perilla leaves are extracted with water under acidic conditions of pH 0.5 to 2.3 and heating conditions of 80 to 93 ° C. , and the resulting extract is adsorbed resin This is a method for producing a phenol-containing extract characterized in that an extract is obtained by chromatography using, and then the extract is concentrated and powdered as necessary.
According to a second aspect of the invention, the acidic conditions, citric acid, lactic acid, malic acid, tartaric acid, succinic acid, The process of claim 1 wherein the acidic conditions with hydrochloric acid or phosphoric acid.
The present invention according to claim 3 is the method according to claim 1 or 2 , wherein the adsorptive resin is an aromatic system or a substituted aromatic system.
The present invention according to claim 4 is the method according to claim 3 , wherein the adsorptive resin has a specific surface area of 500 to 2000 m ² / g and a pore radius of 20 to 200 mm.
The present invention according to claim 5 is the method according to any one of claims 1 to 4 , wherein the phenol is rosmarinic acid.
The present invention according to claim 6 is an extract or powder containing phenols obtained by the method according to any one of claims 1 to 5 .
[0006]
DETAILED DESCRIPTION OF THE INVENTION
In the method of the present invention for producing a phenol-containing extract from salted red perilla leaves, since the extract is also used in the food field, it is first required that the production method is harmless to the human body. The Furthermore, since the raw material is a salted product, a desalting treatment is required, but it is an important issue that the content of the target phenols is not reduced by this treatment.
[0007]
Therefore, in the present invention, a phenol-containing extract is extracted from salted red perilla leaves using water .
[0008]
The extraction is preferably carried out under acidic conditions and under heating conditions. Specifically, for example, citric acid, lactic acid, malic acid, tartaric acid, succinic acid, hydrochloric acid, phosphoric acid and the like are used as the acid, and the pH is adjusted to 0. Extraction is performed under conditions of 5 to 2.3 .
Moreover, as heating conditions, it is 30-93 degreeC, Preferably it is 80-93 degreeC, Usually, 1 to 60 minutes, Preferably 20 to 40 minutes are extracted.
[0009]
Next, a method for extracting a phenol-containing extract from salted red perilla will be described. For example, when extraction is performed using only water as a hydrophilic solvent, water is added to 100 parts by weight of the raw salted red perilla leaf. 100 to 5000 parts by weight, preferably 200 to 1000 parts by weight are used. The acidic conditions, heating conditions, etc. are as described above.
[0010]
Since the extract obtained by filtration after the extraction treatment contains salts and other components in addition to the target phenols, it is necessary to perform a purification operation such as a desalting operation.
As a purification operation for this purpose, chromatography using an adsorptive resin is suitable. That is, after passing the extract through a column packed with an adsorbent resin to selectively adsorb the phenols that are the target components in the extract, the phenols are separated and eluted with a hydrous hydrophilic solvent. Collect.
[0011]
The adsorptive resin used in the present invention is classified into aromatic, substituted aromatic and acrylic based on its chemical structure. The aromatic adsorptive resin is a crosslinked styrene-based porous polymer such as Diaion HP20, Diaion HP21, Sepabead SP825, Sepabead SP850 (all trade names, manufactured by Mitsubishi Chemical Corporation), and the like. It is done. The substituted aromatic-based adsorbent resin is a resin in which a bromine atom is bonded to an aromatic nucleus of an aromatic polymer, and examples thereof include Sepabead SP207 (trade name, manufactured by Mitsubishi Chemical Corporation). The acrylic adsorptive resin has a methacrylic acid ester polymer as a skeleton, and examples thereof include Diaion HP2MG (trade name, manufactured by Mitsubishi Chemical Corporation). Among these adsorptive resins, aromatic and substituted aromatic systems are preferable, and aromatic systems are more preferable.
Furthermore, adsorbent resin has a specific surface area of greater than 500 meters ² / g, preferably they are preferably 500~2000m ² / g, and a pore radius smaller than 200 Å, preferably 20～200A, more preferably 30 to A resin of 150 kg is preferable.
[0012]
Next, regarding the amount of the adsorptive resin used, the volume ratio of the resin amount to the extract is from 1: 1 to 1: 300, preferably from 1: 3 to 1: 150.
By passing the extract through the column filled with the adsorbent resin, the phenols are selectively adsorbed by the resin.
[0013]
After adsorbing phenols to the resin, the non-adsorbing substance is washed away by passing water. The amount of water used at this time is 1 to 20 times, preferably 3 to 10 times the amount of the column resin.
Thereafter, phenols are eluted using a hydrous hydrophilic solvent as an eluent. Here, as the hydrous hydrophilic solvent, for example, a water-ethanol mixed solution such as a 70% aqueous ethanol solution is suitable. Water and ethanol are used in a volume ratio of 10:90 to 80:20, preferably 50:50 to 20:80. The amount of the eluent is 1 to 20 times the amount of the column resin, and 2 to 10 times the preferred amount.
[0014]
Here, phenols contained in red perilla leaves will be described.
Examples of phenols include flavonoids, anthocyans, and phenolic acids. Flavonoids include flavones, isoflavones, flavonols, flavanones, and the like, and examples of anthocyan include cyanidin, delphinidin, shisonin, and glycosides thereof.
[0015]
Examples of the phenolic acid include rosmarinic acid, chlorogenic acid, coumaric acid, caffeic acid, gallic acid, vanillic acid, ferulic acid, protocatecic acid and the like.
In addition to the content of rosmarinic acid in phenolic acids, it is the main active ingredient for antiallergic action.
[0016]
In the present invention, the above eluate (extract) can be used as a red perilla extract as it is, but it is preferable to concentrate and powder as necessary. In this case, the eluate is first concentrated under reduced pressure. Next, freeze drying or spray drying is performed to pulverize the obtained concentrated liquid.
In addition, when spray drying is performed, powdering can be efficiently performed by adding dextrin, lactose, or the like as a binding material. Moreover, it is possible to adjust the phenol content.
[0017]
【Example】
Hereinafter, the present invention will be specifically described with reference to experimental examples, but these do not limit the present invention.
Example 1
After adding 50000 ml of hot water of 90 ° C. and 500 g of citric acid to 5000 g of salted red perilla whole leaves (pH 2.3), stirring at 93 ° C. for 30 minutes and extracting, filtered using No. 6 filter paper, Cooled to room temperature.
An open column (350 ml) filled with 350 ml of an aromatic adsorptive resin (trade name: Sepabead SP850, specific surface area 1000 m ² / g, pore radius 35-45 mm, manufactured by Mitsubishi Chemical Corp.) (Inside diameter 8 cm × length 40 cm), chromatography was performed.
Then, it was washed through 3500 ml of water to remove non-adsorbing components. Next, it was eluted by flowing 3000 ml of 70% ethanol aqueous solution.
[0018]
The obtained eluate was concentrated under reduced pressure to Bx24 and then freeze-dried to obtain 43 g of powder. This powder contained 76.0% by weight of phenols (per weight of powder), and the rosmarinic acid content was 35.5% by weight (per weight of powder).
Phenols were measured by the Prussian blue method (see Methods in Enzymology, vol. 234, 432-433). That is, rosmarinic acid (manufactured by extrasynthese) dissolved in a 1% citric acid solution was obtained from a calibration curve prepared in advance as a standard. The amount of rosmarinic acid was analyzed by HPLC with reference to Planta Medica, 63, 177-179 (1997). That is, the HPLC analysis was carried out in a gradient system using two liquids, liquid A (0.1% trifluoroacetic acid-containing distilled water) and liquid B (0.1% trifluoroacetic acid-containing acetonitrile) as the eluent. . As a column, ODS120T (manufactured by Tosoh Corporation, 4.6 mm × 150 mm) was used, the flow rate was 0.8 ml / min, and detection was performed at UV 320 nm. The gradient conditions are as follows.
[0019]
0-10 min: A liquid / B liquid = 90/10
10-35 min: A liquid / B liquid = 90/10 → A liquid / B liquid = 60/40
35-40 min: A liquid / B liquid = 60/40 → A liquid / B liquid = 0/100
40-45 min: A liquid / B liquid = 0/100
[0020]
Example 2
To 1000 g of salted red perilla leaves, 2000 ml of water and 83 ml of concentrated hydrochloric acid were added (pH 0.5), and the mixture was extracted by stirring at 80 ° C. for 40 minutes, followed by filtration using No. 6 filter paper and cooling to room temperature.
An open column (350 ml) filled with 350 ml of an aromatic adsorptive resin (trade name: Sepabead SP825, specific surface area 1000 m ² / g, pore radius 50-60 mm, manufactured by Mitsubishi Chemical Corp.) (Inside diameter 8 cm × length 40 cm), chromatography was performed.
Thereafter, it was washed with 1200 ml of water to remove non-adsorbing components such as salt. Subsequently, 1000 ml of 80% ethanol aqueous solution was flowed and eluted.
The eluate with an 80% aqueous ethanol solution was concentrated under reduced pressure to Bx24 and then freeze-dried to obtain a powder. This was analyzed in the same manner as in Example 1. As a result, it contained 48.9% by weight of phenols and 23.1% by weight of rosmarinic acid, and the powder weight was 9.2 g.
[0021]
Example 3
After adding 50000 ml of hot water of 90 ° C. and 500 g of citric acid to 5000 g of salted red perilla whole leaves (pH 2.3), stirring at 93 ° C. for 30 minutes and extracting, filtered using No. 6 filter paper, Cooled to room temperature.
An open column filled with 350 ml of an aromatic adsorbent resin (trade name: Sepabead SP850, specific surface area 1000 m ² / g, pore radius 35-45 mm, manufactured by Mitsubishi Chemical Corporation) with the extract thus obtained ( (Inside diameter 8 cm × length 40 cm), chromatography was performed.
Then, it was washed through 3500 ml of water to remove non-adsorbing components. Next, it was eluted by flowing 3000 ml of 70% ethanol aqueous solution.
[0022]
Next, the obtained 70% ethanol eluate was concentrated under reduced pressure to Bx24, and 1000 g of dextrin (trade name: Amicol 6-H, manufactured by Nissho Chemical Co., Ltd.) was added and spray-dried. As a result, 1043 g of red perilla extract powder was obtained.
[0023]
Comparative Example 1
After adding 5000 ml of water and 50 g of citric acid to 500 g of salted red perilla leaves (pH 2.3) and stirring for 30 minutes at 93 ° C., the mixture was filtered using No. 6 filter paper and cooled to room temperature. .
An open column filled with 35 ml of an acrylic adsorptive resin (trade name: Diaion HP2MG, specific surface area 500 m ² / g, pore radius 200-300 mm, manufactured by Mitsubishi Chemical Corp.) (Inside diameter 8 cm × length 40 cm), chromatography was performed.
Thereafter, it was washed with 350 ml of water to remove non-adsorbing components. Next, it was eluted by flowing 300 ml of 70% ethanol aqueous solution.
[0024]
Next, the 70% ethanol eluate was concentrated under reduced pressure to Bx24 and then lyophilized to obtain a powder. This was analyzed in the same manner as in Example 1. As a result, it contained 24.6% by weight of phenols and 11.6% by weight of rosmarinic acid, and the powder weight was 4.4 g.
[0025]
【The invention's effect】
INDUSTRIAL APPLICABILITY The present invention provides a production method that can efficiently recover phenols contained in red perilla using salted red perilla leaves as a raw material. Used as anti-allergic foods and drinks that have the effect of improving symptoms such as hay fever and atopic dermatitis by adding red perilla extract containing phenol itself or concentrated and powdered foods to foods and drinks it can.

Claims (6)

The salted red perilla leaves were extracted with water under acidic conditions of pH 0.5 to 2.3 and heated conditions of 80 to 93 ° C. , and the resulting extract was subjected to chromatography using an adsorptive resin to obtain an extract. Subsequently, the said extract is concentrated and pulverized as needed, The manufacturing method of the phenol containing extract characterized by the above-mentioned. The method according to claim 1 , wherein the acidic condition is an acidic condition with citric acid, lactic acid, malic acid, tartaric acid, succinic acid, hydrochloric acid or phosphoric acid. The method according to claim 1 , wherein the adsorptive resin is an aromatic system or a substituted aromatic system. The method according to claim 3 , wherein the adsorbent resin has a specific surface area of 500 to 2000 m ² / g and a pore radius of 20 to 200 mm. The method according to any one of claims 1 to 4 , wherein the phenol is rosmarinic acid. The extract or powder containing the phenols obtained by the method according to any one of claims 1 to 5 .