KR20160149570A

KR20160149570A - Method for preparing schisandra chinensis comprising lignan in high concentration

Info

Publication number: KR20160149570A
Application number: KR1020150086752A
Authority: KR
Inventors: 이승주; 정승원
Original assignee: 동국대학교 산학협력단
Priority date: 2015-06-18
Filing date: 2015-06-18
Publication date: 2016-12-28

Abstract

More particularly, the present invention relates to a method for producing an extract of Omiza containing a high concentration of lignan, and more particularly, to a method for preparing an extract of Omiza containing high concentrations of lignans by hydrolyzing an organic acid after electrodialysis of the Omiza extract, To a high concentration of lignans which can improve the high value of the omija.

Description

TECHNICAL FIELD [0001] The present invention relates to a method for preparing an extract of Schizandra chinensis L.,

Schizandra chinensis Baillon ( Schizandra chinensis Baillon ) is the main medicinal plant used for raw materials and food raw materials. It is a mature fruit of Schizandra chinensis Baillon such as deciduous wood belonging to the Schizandraceae family.

Omija has five flavors and unusual orientations in sweet, spicy, spicy, spicy, and salty, and has been widely used as an omija tea, Omija liquor, and Omiza lacquer.

The main active ingredients contained in Omiza are gomisin A, B, C, D, E, F, G, K3, N, J, schizandrin A, B, C, schisandrol Lignan compound which contains various components such as oil, essential oil and pigment and has various pharmacological functions including inhibition of hepatotoxicity and is used as a raw material for medicines and food. Recently, studies on the antioxidative and antimicrobial activities of the extracts of omija and lignans have been reported, and it has been reported that the olanzoline has a pharmacological effect such as hypoglycemic action, anti-ulcer action, chronic hepatitis treatment effect, .

The lignans can be classified into four main categories according to their therapeutic properties: adaptogenic action, liver protection, liver stabilization and liver regeneration.

The augotagenic action of lignans has a unique mechanism of action and includes lignans which act as energizers, nervous system promoters, oxygenators, immunomodulators, antioxidants and skin protectants. The lignan has been found to reduce hyperlipemia and significantly inhibit the progression of arteriosclerosis. Lignans is also closely linked to increased levels of ascorbic acid in the liver, inhibition of NADPH oxidation and lipid peroxidation, and the induction of hepatic microsomal cytochrome P-450 enzymes. Liver protection Lignans are known to protect the liver from cholestasis induced by toxic substances. Furthermore, it has been demonstrated that lignan compounds contribute to liver regeneration through induction of hepatocyte growth factor mRNA transcription, promotion of hepatocyte proliferation, and increased blood flow to the liver. It is also known that the above lignan induces an increase in the activity of mitochondrial reductase glutathione and the level of glutathione in mitochondria in order to promote the regeneration ability through synthesis of intercellular glycogen and decrease of glutathione.

As the effectiveness of lignan is proved, the demand for lignan is increasing. Therefore, it is necessary to further study a method of extracting lignan at a high yield, economically and easily.

Korean Patent Laid-Open No. 10-2011-0027339

In order to solve the problems of the prior art as described above, the present invention provides a method for producing lignan, which comprises high concentration of lignan, which is a functional ingredient, by hydrolyzing an enzyme, And a method for producing the same.

Another object of the present invention is to provide a method for preparing an extract of Omiza, which can improve the high added value of Omija and further contains lignans which can be utilized for the development of various functional foods and natural materials.

It is another object of the present invention to provide an Omija extract containing a high concentration of lignans excellent in anticancer, antiviral, anti-inflammatory and vascular protection functions, and an Omija liquid containing the same.

In order to accomplish the above object, the present invention provides a method for preparing an omeprazole extract, And a step of treating a hydrolytic enzyme with an extract of Schizandra chinensis from which the free acid has been separated. The present invention also provides a method for producing an extract of Schizandra chinensis containing a high concentration of lignans.

The electrodialysis can be performed by using at least one electrode solution selected from sodium chloride (NaCl), sodium carbonate (NaCO ₃ ), sodium sulfate (Na ₂ SO ₄ ) and potassium chloride (KCl) at a room temperature under a condition of 10 to 20 V for 50 to 100 minutes And the current density is changed to 50 to 150 A / m < ² >.

The hydrolytic enzyme treatment may be performed by adding 0.01 to 5.0% by weight of β-glucosidase, galactosidase and the like to the total weight of the extract of Omiza, and the treatment is carried out at 25 to 30 ° C. for 1 to 2 hours And the reaction can be carried out.

The present invention also provides an extract of Omija, which is prepared by the above method and has an increase in lignin content by 20% or more as compared with before electrodialysis and hydrolysis.

The present invention also provides an Omija liquid comprising the above-mentioned Omija extract as an active ingredient.

According to the present invention, the extract of Schizandra chinensis can be electrodialyzed, the organic acid can be separated and treated with the hydrolytic enzyme, and the lignan, which is a functional ingredient, can be contained at a high concentration and the absorption of lignans can be promoted. In addition, according to the present invention, it is possible to improve the high added value of Omija and further contribute to the development of various functional foods and natural materials.

Figure 1 shows chromatograms of a standard solution, a control solution and an experimental solution.
2 is a graph showing the results of the calibration curve of the standard solution, seedrazine.
3 is a graph showing a calibration curve of a standard solution of hypermucin A as a standard solution.

Hereinafter, the present invention will be described in detail.

The method of the present invention includes a step of electrodialysis of an extract of Omija to separate an organic acid, and a step of treating a hydrolytic enzyme with an extract of Omija, from which the free acid has been separated. The Omija extract prepared at this stage has a high concentration of lignans do.

Omiza contains a high content of phenolic compounds, and the main component of these phenolic compounds is lignans, which are present in the form of glycosides linked to some sugars. Accordingly, in the present invention, a large amount of lignan can be separated by separating a phenol-based glycoside into a sugar and an aglycone through a hydrolytic enzyme treatment to prepare an extract of Omiza containing lignans in high purity. In addition, the present invention can maximize the hydrolytic enzyme treatment effect by inhibiting the enzyme activity inhibition at low pH by separating the organic acid in the omiza by electrodialysis prior to the hydrolytic enzyme treatment.

The Omiza extract used for the electrodialysis of the present invention can be extracted by a conventional method. As the extraction solvent, distilled water or a lower alcohol such as ethanol, methanol, butanol, acetone, ethyl acetate, dichloromethane, or an intermediate polar organic solvent may be used alone or as a mixture of two or more kinds. In particular, It is better to extract it.

Specifically, the above-mentioned Omiza extract was mixed with water at a weight ratio (w / v) of 1: 10 with respect to the weight of dried omija, and then extracted at 80 ° C for 3 hours or extracted with ethanol, methanol, butanol, acetone, ethyl acetate, dichloromethane Of lower alcohol or an intermediate polar organic solvent as an extraction solvent, and extracting solvent to the weight of dried omija at a weight ratio (w / v) of 1:10, and then extracting at room temperature for 24 hours. The extract can be prepared by a conventional extraction method such as a low temperature extraction method, a room temperature extraction method, an ultrasonic extraction method, or a reflux extraction method. The resulting extract is filtered with a filter paper, and the obtained filtrate is freeze-dried, .

The above-extracted Omiza extract can be separated into high purity through various conventional separation methods as necessary. Separation methods such as solvent distribution, precipitation separation, recrystallization, and column chromatography can be used as separation methods usable at this time.

The thus-prepared Omiza extract is neutralized to have a pH of about 5.5. At this time, the neutralization process may be performed using 0.1N NaOH in the extract of Omija, or 2mL of 1 mM NaNO ₂ solution may be added, and the treatment may be performed using 0.1N HCl and 0.2M citrate buffer.

Then, the above-mentioned Omija extract is subjected to electrodialysis to separate the organic acid contained in the Omija.

The electrodialysis can be performed using a conventional electrodialyzer. Specifically, the electrodialysis can be performed using an electrodialyzer in which a cation exchange membrane and an anion exchange membrane usable for food are arranged in order, at room temperature, at 10 to 20 V for 50 to 100 minutes And the current density can be changed by 50 to 150 A / m < ² >. At this time, an aqueous sodium chloride solution can be added to the electrodialyzer so that initial electricity flows well. As the electrode solution, at least one selected from sodium chloride (NaCl), sodium carbonate (NaCO ₃ ), sodium sulfate (Na ₂ SO ₄ ) and potassium chloride (KCl) can be used. In particular, a solution of sodium carbonate (NaCO ₃ ) It is good.

Electrodialysis is a process for ion-exchanging, purifying, and concentrating an electrolyte present in an aqueous solution by passing ions through an ion exchange membrane using a potential difference. The electrodialysis process facilitates the production of high purity organic acids by not only separating the organic acids from the fermentation broth but also reducing the cost of energy required for the concentration of organic acids in the lower process. In addition, electrodialysis is a very eco-friendly process because it does not generate any impurities caused by the process, unlike a general refining process.

The main parameters affecting the separation process of organic acids in electrodialysis are the current density, the concentration of the raw solution, and pH. The electrodialysis condition in the present invention is a principle of converting a salt-like substance into an acid and an alkali form using a bipolar membrane. The hydrogen ions and the hydroxide ions generated on both surfaces of the bipolar membrane are transferred to the feed compartment and the permeate compartment, respectively. In order to match the ion concentration in the compartments, the ions having an inner charge pass through the ion exchange membrane and move to the other compartment. The generated acid is accumulated.

Therefore, in the present invention, by using the electrodialysis method as described above, it is possible to effectively separate (remove) the organic acid contained in the omisza at low cost.

The degree of separation (removal) of the organic acid by electrodialysis can be confirmed by measuring the electrical conductivity and pH.

When the electrodialysis is performed as described above, the organic acid contained in the omija can be separated, and the omija extract in which the organic acid is separated can maintain the pH above a certain level, thereby inhibiting the hydrolytic enzyme activity inhibition due to the low pH. Therefore, if the electrodialysis is performed, the effect of the hydrolytic enzyme treatment can be further maximized. In addition, since the electrodialysis process does not perform chemical neutralization, the problem of metal ions remaining from the alkali substance used in neutralization can also be prevented through the electrodialysis process.

As described above, hydrolytic enzymes are applied to the extract of Omija, which is obtained by separating organic acids through electrodialysis.

The hydrolytic enzymes can be used without limitation as long as they are enzymes capable of separating glycosides from sugar and aglycons. Specifically, beta-glucosidase, galactosidase and the like are used .

The hydrolytic enzyme is added in an amount of 0.01 to 5.0% by weight, preferably 0.1% by weight, based on the total weight of the extract, and the reaction is preferably carried out at 25 to 30 ° C for 1 to 2 hours. When the hydrolytic enzyme is added in an amount of less than 0.01% by weight, the hydrolysis efficiency may be insufficient. When the hydrolytic enzyme is added in an amount exceeding 5.0% by weight, the hydrolysis efficiency is not significantly changed.

After the hydrolytic enzyme treatment, the extract of Omiza increased the lignin content by 20% or more as compared with that before the electrodialysis and hydrolytic enzyme treatment, thereby preparing an Omija extract having enhanced anticancer, antiviral, anti-inflammatory and vascular protection functions .

The present invention also provides an Omija liquid comprising the above-prepared Omija extract as an active ingredient.

According to the present invention, the above-mentioned Omiza liquid may further comprise an Omija water extract together with an Omiza extract containing a high concentration of lignan as a functional ingredient.

Preferably, the Omiza extract is contained in an amount of 2 to 10% by weight (v / v), more preferably 5% by weight, based on the total volume of the omithine solution. If the content is less than 2% by weight, the functional effect of lignans may be insignificant. If the content is more than 10% by weight, the effect of lignan relative to the usage amount is relatively low and there is no significant change in sensory characteristics and functionality, May lead to an increase in product prices. In particular, when added at 5% by weight based on the total volume of the omiza solution, the above-mentioned Omiza extract is superior in sensory properties and most effective in antibacterial activity.

In addition, the above-mentioned Omija hot-water extract may be prepared by extracting water at 30-40 ° C. with water by an ordinary method according to a conventional method.

Hereinafter, the present invention will be described in more detail with reference to examples. These embodiments are for purposes of illustration only and are not intended to limit the scope of protection of the present invention.

Example 1

In September, 2012, one of the omija harvested at Mungyeong was selected to have a size of 0.7-0.9 mm and dried at 20 ° C at low temperature. 1,100 ml of distilled water was added to 110 g of the dried omija as an extraction solvent and extracted at 30 ° C to prepare an Omija extract (pH 2.7).

The pH of the Omiza extract was adjusted to 5.5, and 500 ml of Omiza extract was placed in an electrodialyzer (MicroAcylizer-S3, Asahi Chemical Co., Shizuoka, Japan) for electrodialysis at room temperature and 12V for 180 minutes. At this time, the electrodialyzer used an electrodialyzer in which five cation exchange membranes (AMX-SB) and anion exchange membranes (CMX-SB) were arranged in order. In the concentrating chamber of the electrodialyzer, 500 ml of 0.5% sodium chloride aqueous solution was added for initial current flow, and 500 ml of 5% sodium carbonate solution was used as the electrode solution. The electroconductivity was measured to determine the degree of elimination of neutralized organic acid, and it was confirmed that 95% or more of the organic acid contained in the extract of Omija was separated, and the electrodialysis was terminated.

After the electrodialysis, 0.1% by weight of β-glucosidase and galactosidase were added to the Omiza extract, respectively, and the mixture was reacted at 25 to 30 ° C. for 1 to 2 hours for hydrolytic enzyme treatment.

Experimental Example 1. Determination of Organic Acid Separation by Electrodialysis

The electrical conductivity and organic acid were measured in order to confirm the degree of separation of organic acid by the electrodialysis process in the Example 1.

First, in Example 1, the extract of Omiza before electrodialysis was filtered using Watson filter paper (20μ), and the electrical conductivity and pH immediately after extraction were measured using an electric conductivity meter. Thereafter, the pH of the Schizosacralis japonica extract was adjusted to 5.5, and then the electrical conductivity and pH were measured. After completion of the electrodialysis, the electrical conductivity and pH were measured, and the results are shown in Table 1 below.

In addition, 0.1 M sodium hydroxide solution was titrated in 10 ml of the extract using an Omija extract before electrodialysis and an Omija extract after completion of electrodialysis until the pH reached 8.3, and the acidity was measured according to the following formula (1). The results are shown in Table 2 below.

[Equation 1]

sample Electrical conductivity (mS / cm) Immediately after extraction, Omija extract 3.26 Omija extract at pH 5.5 18.24 Omija extract after electrodialysis 1.16

sample pH Acidity (% citrate) Immediately after extraction, Omija extract 2.67 0.0219 After electrodialysis 5.5 0.0012

As shown in Tables 1 and 2, when the electrodialysis process was performed prior to the hydrolytic enzyme treatment of Omiza extract according to the present invention, the electrical conductivity fell from 17.58 mS / cm to 1.62 mS / cm, , And the acidity of the extract of Omija was 0.0219, which was about 0.0012 after electrodialysis, indicating that about 95% of the organic acid was isolated.

Experimental Example 2. Measurement of Lignan Content

In order to measure the content of lignans contained in the extract of Omija after the electrodialysis and hydrolytic enzyme treatment in Example 1, the following experiment was conducted.

First, the hydrolytic enzyme-treated Omija extract of Example 1 was subjected to electrodialysis and then lyophilized and powdered. 500 mg of the powder was dissolved in 20 ml of methanol, sonicated for 20 minutes and filtered. After filtration, 20 ml of methanol was added to the residue, and the mixture was subjected to ultrasonic extraction for 20 minutes and filtration. The filtrate was combined and the filtrate was adjusted to 50 ml with methanol and filtered through a 0.45 μm nylon membrane syringe filter.

As the reference material, the superficial lignin A and sizindrin were dissolved in methanol (Duksan, HPLC grade), and the stock solution was appropriately diluted with methanol and used as a standard solution. As a control, the omija extract prepared in Example 1 was lyophilized and powdered. 500 mg of the powder was dissolved in 20 ml of methanol, sonicated for 20 minutes and filtered. After filtration, 20 ml of methanol was added to the residue, and the mixture was subjected to ultrasonic extraction for 20 minutes and filtration. The filtrate was combined and the filtrate was adjusted to 50 ml with methanol and filtered through a 0.45 μm nylon membrane syringe filter.

C18 column. High glomerulin A and sizzydrine were detected in the control and experimental groups at 254 nm. High myrin A and sizzardine content in the samples were measured using a standard curve. The analysis was performed according to the conditions shown in Table 3 below.

Instrument Agilnet 1260 Infinity Detector PDA detector wavelength 254 nm column Shiseido Capcell PAK C18 MG ((4.6 mm x 250 nm, 5 m) Mobile phase Acetonitrile: DW: formic acid = 70: 30: 0.1 flux 0.6 ml / min Dose 10 μl Oven temperature 30 ℃

The chromatograms of the standard solution, the control solution and the test solution are shown in FIG. 1, and the calibration curve results of the standard solution are shown in Table 4, FIGS. 2 and 3, and the control solution and the experimental solution Of the present invention are shown in Tables 5 and 6. < tb > < TABLE > The following results were obtained through five repeated experiments.

Sizandrin The superstition A STD level Concentration (/ / ml) area STD level Concentration (/ / ml) area One 0.103 3.21551 One 0.145 3.68673 2 0.206 6.32714 2 0.291 752706 3 0.825 25.94334 3 1.1625 3115882 4 3.3 103.57177 4 4.65 123.6125 5 6.6 211.73378 5 9.3 253.16393

&Quot; (2) "

As shown in Tables 5 and 6, in the control group without electrodialysis and hydrolytic enzyme treatment, the content of sizzardine was 0.056 ± 0.0005 mg / g, and the content of hypermucin A was 0.11 ± 0.0002 mg / g. On the other hand, in the experimental group of Example 1 in which hydrolytic enzyme treatment was performed according to the present invention, the sizzolidine content was 0.067 ± 0.0039 mg / g and the hypericin A content was 0.013 ± 0.0007 mg / g. From these results, it can be seen that the hydrolysis enzyme-treated Omiza extract according to the present invention increased the lignan content by about 20% as compared with the Omiza extract without electrodialysis and hydrolytic enzyme treatment.

Although the present invention has been described in terms of the preferred embodiments mentioned above, it is possible to make various modifications and variations without departing from the spirit and scope of the invention. It is also to be understood that the appended claims are intended to cover such modifications and changes as fall within the scope of the invention.

Claims

Separating the organic acid by electrodialysis of the extract of Omija; And
Treating the hydrolytic enzyme with an extract of Schizandra chinensis isolated from the free acid;
Wherein the lignan is present at a high concentration.

The method according to claim 1,
The extract of Omiza is prepared by adding at least one solvent selected from the group consisting of distilled water, ethanol, methanol, butanol, acetone, ethyl acetate and dichloromethane to a weight ratio of dried seaweed to a dry weight ratio of 1:10 (w / v) A method for producing an Omija extract containing a high concentration of lignans.

The method according to claim 1,
The electrodialysis can be performed by using at least one electrode solution selected from sodium chloride (NaCl), sodium carbonate (NaCO ₃ ), sodium sulfate (Na ₂ SO ₄ ) and potassium chloride (KCl) at a room temperature under a condition of 10 to 20 V for 50 to 100 minutes Wherein the current density is changed in a range of 50 to 150 A / m < ² >.

The method according to claim 1,
Wherein the hydrolytic enzyme is any one selected from the group consisting of beta-glucosidase, galactosidase, and mixtures thereof.

The method according to claim 1,
Wherein the hydrolytic enzyme treatment is carried out by adding 0.01 to 5.0% by weight based on the total weight of the Schizandra chinensis extract and reacting at 25 to 30 ° C for 1 to 2 hours to prepare an omija extract containing high concentrations of lignans .

The method according to claim 1,
Wherein the extract of Omiza after the hydrolytic enzyme treatment has a lignan content increased by at least 20% as compared with that before treatment with electrodialysis and hydrolytic enzymes.

An extract of Omija produced by the method of any one of claims 1 to 6.

An omiza solution characterized by comprising an extract of Omiza prepared by the method of any one of claims 1 to 6 as an active ingredient.

9. The method of claim 8,
Wherein the Omiza extract is contained in an amount of 2 to 10% by weight (v / v) based on the total volume of the Omija liquid.