KR20210073869A - Method of preparing juniper extract containing polyphenol - Google Patents

Abstract

The present invention relates to a method for preparing a juniper extract containing polyphenols, and more particularly, to a method for preparing a juniper extract containing a high content of polyphenols by increasing a polyphenol content of the juniper tree by using supercritical water, subcritical water, and oxidation reaction. The present invention provides a method for preparing a polyphenol-containing juniper extract by using eco-friendly subcritical water and supercritical water, thereby replacing a risky conventional organic solvent extraction and providing an extraction method with secured stability, and providing an effect of achieving excellent polyphenol extraction efficiency (process time, polyphenol content) compared to hot water extraction. In addition, it is possible to improve a polyphenol conversion rate of phenolic compounds in the juniper extract through an additional oxidation reaction, thereby achieving an effect of providing an environmentally friendly hydrolysis process not involving the generation of neutralized salts and residual salts due to the conventional acid/base reaction.

Description

Method of preparing juniper extract containing polyphenol {Method of preparing juniper extract containing polyphenol}

The present invention relates to a method for producing a juniper extract containing polyphenols, and more particularly, by using supercritical water, subcritical water, and oxidation reactions to increase the polyphenol content of juniper trees, thereby providing a high content of polyphenols. It relates to a method for preparing a juniper extract containing

Reactive oxygen species (ROS) generated by intracellular metabolic processes cause damage to biomolecules including DNA, and continuous oxidative damage causes chronic chronic diseases such as cancer, arthritis, aging, autoimmune disorders, and cardiovascular diseases. It acts as a cause of diseases and degenerative diseases. Substances that can prevent such oxidative damage include antioxidants, and polyphenols, which are secondary metabolites of plants, are one of the representative antioxidants that stabilize free radicals of ROS through hydroxyl substitution.

On the other hand, juniper, which has been used in the private sector for a long time, is called dupine in oriental medicine and is used for the treatment of sweating, diuresis, neuralgia, and rheumatism. The pharmacological action of juniper extract is known to have gastric action, expectorant action, antibacterial action, antifungal action, antirheumatic action, diuretic action, antiseptic action, sedative action, anti-inflammatory action, and sterilization action. Cocci, Escherichia coli, Typhus, Shigella and the like are known. In order to use the active ingredient of the juniper tree, the active ingredient is extracted from the juniper tree using water, a solvent, a mixed solvent, etc., or a juniper extract containing the active ingredient is prepared and used.

Accordingly, 'Korea Patent Registration No. 10-0941133' relates to caprylyl glycol ethylhexylglycerin having antiseptic and anti-inflammatory activity, juniper extract, leek extract and perilla extract, and a cosmetic composition containing the same as active ingredients. It discloses a juniper extract obtained by using an organic solvent such as acetone, ethyl acetate, chloroform, etc. as a constituent of a cosmetic composition, but this organic solvent is a raw material for products directly related to stability such as cosmetics or food. There is a problem in that it is difficult to secure stability to use as a composition, and when using such a composition, it may appear as a problem of various physical disorders after a long period of time.

In general, active ingredients of natural products have the characteristic of being more efficiently dissolved in organic solvents, so there is an urgent need for a method for extracting natural products that guarantees stability while ensuring the same or high extraction efficiency as those organic solvents.

Therefore, in the present invention, water is used as a solvent to secure stability and extraction efficiency while replacing the conventional organic solvent, and at the same time, hydrolysis of supercritical water, extraction of subcritical water, and oxidation reaction of the extract have a high content of polyphenol, an excellent antioxidant An object of the present invention is to provide a method for preparing a juniper extract containing

KR 10-0941133 B1 KR 10-2012-0122509 A

In order to solve the above problems, an object of the present invention is to provide a method for producing a juniper extract using subcritical water, which is eco-friendly and secure, and has high extraction efficiency of active ingredients.

Another object of the present invention is to provide a method for preparing a polyphenol-containing juniper extract using supercritical water hydrolysis and subcritical water extraction.

The present invention in order to solve the above object,

preheating purified water to form pressurized hot water;

converting the pressurized hot water into a subcritical state to form subcritical water;

extracting the juniper tree with the subcritical water to form a juniper subcritical water extract; and

It provides a method for producing a polyphenol-containing juniper extract comprising; filtering and deodorizing the juniper subcritical water extract.

The present invention in order to solve the other object,

preheating purified water to form pressurized hot water;

converting the pressurized hot water into a supercritical state to form supercritical water;

hydrolyzing the juniper tree with the supercritical water to form a juniper hydrolyzate;

extracting the juniper hydrolyzate with subcritical water to form a juniper subcritical water extract; and

It provides a method for producing a polyphenol-containing juniper extract comprising; filtering and deodorizing the juniper subcritical water extract.

The present invention provides a method for producing a polyphenol-containing juniper extract using eco-friendly subcritical water and supercritical water, thereby replacing the conventional organic solvent extraction with risk and providing a secure extraction method, Polyphenol extraction efficiency (process time, polyphenol content) is superior to hot water extraction.

In addition, it is possible to improve the polyphenol conversion rate of phenolic compounds in the juniper extract through an additional oxidation reaction, and environmentally friendly hydrolysis that does not involve the generation of neutralized salts and residual salts due to the conventional acid/base reaction There is an effect that can provide a process.

1 is a graph showing the change in permittivity and change in polarity according to the temperature of water.
Figure 2 shows a manufacturing process diagram of juniper polyphenol extract using subcritical water extraction according to an embodiment of the present invention.
Figure 3 shows a manufacturing process diagram of juniper polyphenol extract using supercritical water hydrolysis and subcritical water extraction according to an embodiment of the present invention.
Figure 4 shows a manufacturing process diagram of juniper polyphenol extract using supercritical water hydrolysis, subcritical water extraction and oxidation reaction according to an embodiment of the present invention.
Figure 5 is a structural diagram of the extraction apparatus used in the manufacture of juniper polyphenol extract according to an embodiment of the present invention.

In the present specification, when a part "includes" a certain component, this means that other components may be further included rather than excluding other components unless otherwise stated. And the terminology used in this specification is for describing the embodiments, and is not intended to limit the present invention. As used herein, the singular also includes the plural, unless the phrase specifically states otherwise.

The present invention relates to a method for producing a juniper extract containing polyphenols.

According to one aspect, the present invention comprises the steps of preheating purified water to form pressurized hot water; converting the pressurized hot water into a subcritical state to form subcritical water; extracting the juniper tree with the subcritical water to form a juniper subcritical water extract; and filtering and deodorizing the juniper subcritical water extract; provides a method for producing a polyphenol-containing juniper extract comprising a.

The step of forming subcritical water in this manufacturing method may form subcritical water at a temperature range of 300 to 350 °C, preferably at a temperature of 320 °C to 340 °C. When the temperature is less than 300℃, the dielectric constant of subcritical water is high, making it difficult to replace organic solvents such as ethanol, methanol, and ethyl acetate because it is not suitable for extracting medium polar substances. The thermal power is lowered, and when subcritical water is extracted, the solubility of the oil component is reduced, which may lower the extraction yield. In addition, when the temperature is more than 350 ℃, the dielectric constant of water is significantly lowered as close to the critical temperature range is converted to a non-polar solvent phase may not be suitable for polyphenol extraction of the present invention.

In the method for producing a polyphenol-containing juniper extract of the present invention, the extraction pressure of subcritical water is preferably adjusted in the range of 100 to 150 bar. When the pressure is less than 100 bar, the subcritical water temperature section is a lower pressure section compared to the high temperature section of 300 ° C. Due to the low boiling point increase, it may be difficult to prevent oxidation and carbonization of extracts containing polyphenols during extraction of subcritical water. In addition, when the pressure is more than 150 bar, the density of subcritical water increases and solubility increases, while the rate of material diffusion in the target sample decreases, thereby reducing the overall extraction rate, and the dielectric constant of subcritical water is significantly lowered. Corresponding to the pressure, the extraction performance of the non-polar material is improved than the extraction performance of the intermediate polar material, so it may not be suitable for extracting the polyphenol which is the intermediate polar material desired in the present invention.

In the extraction process, the use of a solvent having a polarity similar to the polarity of the material to be extracted can maximize the extraction yield by increasing solubility. Referring to FIG. 1, water at atmospheric pressure has a high melting point or boiling point due to very strong hydrogen bonding between molecules, and has a high dielectric constant, ε, which is an electrical property of the material (ε>80, at 25℃) polarity Extraction of the compound can be easily performed. In addition, water has properties such as surface tension, viscosity, permittivity, density, and solubility that change with increasing temperature and pressure, so it is effective for extraction of low- or non-polar compounds such as phenolic compounds as well as polar compounds according to the change in characteristics. can be used

The extraction process is the most important process in obtaining physiologically active substances from natural products, and Subcritical Water Extraction (SWE), which extracts using only pure water, changes the relative permittivity, which is the physicochemical property of water, according to polarity. It is a very effective technique that selectively dissolves bioactive substances and facilitates concentration operation. Water is inexpensive, non-toxic, and can be safely disposed of after use, and its properties can be changed simply by temperature and pressure control, so it can be used in various ways for extracting active ingredients depending on the purpose. It is effective in extracting non-polar active ingredients due to the relative dielectric constant.

Subcritical water (SW) is water that remains liquid under pressure (<22.1 MPa) between its boiling point (100°C) and critical temperature (374°C). Pressurized hot water Or superheated water. Compared to conventional reflux cooling extraction, subcritical water extraction (SWE) has a shorter extraction time, consumes a small amount of solvent, and is selective for physiologically active substances by controlling the polarity of the solvent widely through temperature and pressure control. extraction may be possible.

Among the methods for producing a polyphenol-containing juniper extract of the present invention, the method for producing a juniper extract using subcritical water may proceed as shown in the process diagram shown in FIG. 2 , and includes a raw water supply unit, a purified water forming unit, a purified water preheating unit, a subcritical and It can be extracted using the extraction apparatus shown in FIG. 5 including a supercritical conversion device unit, a hydrolysis and extraction unit, an extract purification unit, an extract recovery unit, and a control unit.

Preferably, purified water may be formed by filtering and purifying the raw water supplied from the raw water supply unit through the purified water forming unit, and the purified water may be preheated through the purified water preheating unit to form pressurized hot water. The pressurized hot water can be transferred to the subcritical and supercritical conversion device unit to convert it to a subcritical state, and the subcritical water and juniper can be reacted for subcritical extraction through the hydrolysis and extraction unit. Thereafter, the extracted subcritical water extract may be filtered and deodorized through the extract purification unit, and then recovered through the extract recovery unit.

According to another aspect, the present invention comprises the steps of preheating purified water to form pressurized hot water; converting the pressurized hot water into a supercritical state to form supercritical water; hydrolyzing the juniper tree with the supercritical water to form a juniper hydrolyzate; extracting the juniper hydrolyzate with subcritical water to form a juniper subcritical water extract; and filtering and deodorizing the juniper subcritical water extract; provides a method for producing a polyphenol-containing juniper extract comprising a.

Forming the supercritical water in this manufacturing method may form the supercritical water at a temperature range of 374 to 425 °C, preferably at a temperature of 380 °C to 400 °C. The critical temperature of supercritical water is 374°C, and it can exhibit a characteristic that the number of hydroxide ions greatly increases from the critical temperature. The supercritical water hydrolysis reaction induces the elution action of the cell wall of hydroxide ions, and a temperature of at least 374°C may be required to carry out this reaction. When the formation temperature of supercritical water exceeds 425 ℃, the hydroxyl ion ratio of water to organic substances is very high, which shows the decomposition ability to dissociate most ions and covalent bonds to hydrocarbon compounds. It may be difficult to carry out the hydrolysis reaction of the present invention.

In the method for producing a polyphenol-containing juniper extract of the present invention, the forming pressure of supercritical water is preferably adjusted in the range of 221 to 300 bar. The critical pressure of supercritical water is 221 bar, and since it is a critical pressure that can provide the heat capacity required to rise to the critical temperature of water, it may be difficult to form supercritical water at a pressure of 221 bar or less. In addition, when the pressure exceeds 400 bar, the density of supercritical water is increased, so that the solubility of the non-polar material increases while the material diffusion rate decreases, thereby causing a retrograde phenomenon that can lower the overall hydrolysis rate.

Among the methods for producing a polyphenol-containing juniper extract of the present invention, the method for preparing a juniper extract using supercritical water and subcritical water may proceed as shown in the process diagram shown in FIG. 3 , and a raw water supply unit, a purified water forming unit, and a purified water preheating unit , can be extracted using the extraction device shown in Figure 5, including subcritical and supercritical conversion device unit, hydrolysis and extraction unit, extract purification unit, extract recovery unit, and control unit.

Preferably, purified water may be formed by filtering and purifying the raw water supplied from the raw water supply unit through the purified water forming unit, and the purified water may be preheated through the purified water preheating unit to form pressurized hot water. The pressurized hot water can be transferred to the subcritical and supercritical conversion device unit to convert to a supercritical state, and hydrolysis of juniper can be performed with supercritical water through the hydrolysis and extraction unit, and the subcritical and supercritical conversion device Subcritical extraction can be carried out in the extraction unit by reacting the hydrolyzed juniper with the water converted to the subcritical state through the division. Thereafter, the extracted subcritical water extract may be filtered and deodorized through the extract purification unit, and then recovered through the extract recovery unit.

The method for producing a polyphenol-containing juniper extract of the present invention may preferably use supercritical water, subcritical water and oxidation reaction. By further including an oxidation reaction in which the juniper extract is subjected to an oxidation reaction under an oxygen atmosphere gas, the oxidation reaction of phenolic compounds in the juniper extract can be further accelerated, thereby increasing the amount of polyphenol produced in the extract.

The method for preparing a polyphenol-containing juniper extract using supercritical water, subcritical water and oxidation reaction includes the steps of preheating purified water to form pressurized hot water; converting the pressurized hot water into a supercritical state to form supercritical water; hydrolyzing the juniper tree with the supercritical water to form a juniper hydrolyzate; extracting the juniper hydrolyzate with subcritical water to form a juniper subcritical water extract; oxidizing the juniper subcritical water extract to form a juniper oxidation reactant; and filtering and deodorizing the juniper oxidation reactant.

At this time, the step of forming the oxidation reactant may be performed in a temperature range of 135 to 180 ° C. When the temperature is less than 135 ° C, the memalad reaction of the phenolic compound may not occur, and when the temperature is greater than 180 ° C, the extract Carbonization of hydrocarbon compounds that are vulnerable to heat contained within may be induced.

In addition, the oxidation reaction time may preferably be 10 to 30 minutes, and when the reaction time is less than 10 minutes, the Maillard reaction of the phenolic compound may not occur, and when the reaction time is more than 30 minutes, a significant effect does not appear. it may not be

Among the methods for producing a polyphenol-containing juniper extract of the present invention, the method for preparing a juniper extract using supercritical water, subcritical water and oxidation reaction may proceed as shown in the process diagram shown in FIG. 4, and includes a raw water supply unit, a purified water forming unit, Purified water preheating unit, subcritical and supercritical conversion device unit, hydrolysis and extraction unit, oxidation reaction unit, extract purification unit, extract recovery unit, oxygen gas supply unit and control unit to be extracted using the extraction device shown in FIG. can

Preferably, purified water may be formed by filtering and purifying the raw water supplied from the raw water supply unit through the purified water forming unit, and the purified water may be preheated through the purified water preheating unit to form pressurized hot water. The pressurized high-temperature water can be transferred to the subcritical and supercritical conversion device unit to convert it to a supercritical state, and hydrolysis of juniper can be performed with supercritical water through a supercritical hydrolysis reaction inside the hydrolysis and extraction unit. , it is possible to perform subcritical extraction of the noganju supercritical water hydrolyzate by supplying the subcritical water converted through the subcritical and supercritical conversion device to the inside of the extraction unit. Thereafter, the oxidation reaction of the subcritical water extract can be performed by supplying oxygen gas from the oxygen gas supply unit into the oxidation reaction unit, and the extract can be transferred to the extract recovery unit through the extract purification unit to obtain the juniper extract.

In the present invention, hydrolysis of juniper using supercritical water is a process for improving extraction efficiency by breaking the cell wall and dissolving the eluted phenolic compound in subcritical water, and may be included in the manufacturing method of the juniper extract of the present invention The oxidation reaction is a process for obtaining a juniper extract containing a high concentration of polyphenols by improving the polyphenol conversion of phenolic compounds by promoting the oxidation reaction at a specific temperature and pressure under an oxygen atmosphere gas. It may be included in the preparation method of the extract.

The present invention will be described in more detail by the following examples. However, the following examples are intended to illustrate the present invention, and the scope of the present invention is not limited by the examples. It is provided to complete the disclosure of the present invention, and to completely inform those of ordinary skill in the art to which the present invention pertains to the scope of the invention.

<Example>

Example 1 - Preparation of Juniper Polyphenol Extract Using Subcritical Water Extraction

Juniper tree using subcritical water in the same process as the extraction process diagram shown in FIG. A polyphenol extract was prepared. After purifying raw water with a purified water filtration device using a pressurized pump (pressure 8bar, flow rate 30ml/min) and transferring the purified water to the pre-heater, The purified water was pre-heated (120℃, 8bar) to form pressurized hot water, and the pressurized hot water was transferred to the subcritical water phase conversion device using a high-pressure metering pump to convert to subcritical water (300℃, 100bar) state. did. Thereafter, the metering valve between the subcritical water conversion device and the extractor was gradually opened and closed to maintain the phase equilibrium of the extractor internal pressure and temperature to a subcritical water state (300° C., 100 bar), and then subcritical water hydrolysis was performed for 30 minutes. Next, open and adjust the back pressure regulator installed between the extractor and the micro filter to set the back pressure to 100 bar (At this time, the subcritical water extraction temperature is maintained at 300 °C, and the internal pressure of the reactor is set at 100 bar. oil), the juniper powder charged inside the reactor was extracted using subcritical water for 30 minutes, and the extracted extract in the reactor was filtered through a microfilter and then passed through a deodorizing filter to undergo a deodorization process and then extract It was recovered in the recovery tank.

Example 2 - Preparation of Juniper Polyphenol Extract Using Supercritical Water and Subcritical Water Extraction

Juniper tree using supercritical water and subcritical water in the same process as the extraction process diagram shown in FIG. A polyphenol extract was prepared. After purifying raw water with a purified water filtration device using a pressurized pump (pressure 8bar, flow rate 30ml/min) and transferring the purified water to the pre-heater, The purified water was preheated (120℃, 8bar) to form pressurized hot water, and the pressurized hot water was transferred to the supercritical water phase conversion device using a high-pressure metering pump and converted to supercritical water (420℃, 240bar) state. did. Thereafter, the metering valve between the supercritical water conversion device and the extractor was slowly opened and closed to maintain the phase equilibrium in the supercritical water state (420° C., 240ba) of the internal pressure and temperature of the extractor, and then supercritical water hydrolysis was performed for 30 minutes. Next, the back pressure was set to 100 bar by opening and closing the back pressure regulator mounted between the extractor and the micro filter, and the subcritical water extraction temperature was maintained at 300 °C, and the internal pressure of the reactor was maintained at 100 bar. did. The juniper powder charged inside the reactor was extracted using subcritical water for 30 minutes, and the extracted extract in the reactor was filtered through a microfilter and then passed through a deodorizing filter to undergo a deodorization process and then recovered to the extract recovery tank. did.

Example 3 - Preparation of Juniper Polyphenol Extract Using Supercritical Water, Subcritical Water, and Oxidation Reaction

Juniper using supercritical water, subcritical water, and oxidation reaction in the same process as the extraction process diagram shown in FIG. A polyphenol extract was prepared. After purifying raw water with a purified water filtration device using a pressurized pump (pressure 8bar, flow rate 30ml/min) and transferring the purified water to the pre-heater, The purified water was preheated (120℃, 8bar) to form pressurized hot water, and the pressurized hot water was transferred to the supercritical water phase conversion device using a high-pressure metering pump and converted to supercritical water (420℃, 240bar) state. did. Thereafter, the metering valve between the supercritical water conversion device and the extractor was slowly opened and closed to maintain the phase equilibrium in the supercritical water state (420° C., 240ba) of the internal pressure and temperature of the extractor, and then supercritical water hydrolysis was performed for 30 minutes. Next, the back pressure was set to 100 bar by opening and closing the back pressure regulator mounted between the extractor and the micro filter, and the subcritical water extraction temperature was maintained at 300 °C, and the internal pressure of the reactor was maintained at 100 bar. did. The juniper powder charged inside the reactor was extracted using subcritical water for 30 minutes, and the extracted extract in the reactor was then filtered through a microfilter and passed through a deodorizing filter to go through a deodorization process and then to the extract recovery tank. recovered. The extract transferred to the recovery tank was oxidized under oxygen atmosphere gas at 150° C. and 20 bar for 15 minutes to promote the oxidation reaction of phenolic compounds in the extract to prepare a juniper extract with increased polyphenol production.

Comparative Example 1 - Preparation of juniper polyphenol extract using hot water extraction

After purifying raw water with a purified water filtration device using a pressurized pump (pressure 8bar, flow rate 30ml/min) and transferring the purified water to the pre-heater, Purified water was preheated (120° C., 8 bar) to form hot water. Afterwards, the back pressure regulator installed between the extractor and the micro-filter was opened and closed to set the back pressure to 8 bar, and the hot water extraction temperature was maintained at 100 ° C to extract the juniper powder charged inside the reactor. (The internal pressure of the reactor is maintained at 8 bar) After filtering the extraction residue in the micro filter, it is passed through a deodorizing filter to undergo a deodorization process, and then recovered to the extract recovery tank.

In order to facilitate the description in the specification, the juniper extracts of Examples 1 to 3 are named A, B, and C, respectively, and the juniper extract of Comparative Example 1 is named D.

<Experimental example>

Experimental Example 1 - Analysis of total polyphenol content

In order to analyze the total polyphenol content of juniper extracts according to Examples and Comparative Examples, a color development method using Folin-Ciocalteu phenol reagent was used. After mixing 100 μL of each extract and 2 mL of 2% sodium carbonate solution and reacting for 3 minutes, 100 μL of 50% Paulinciocalto phenol reagent was added and reacted for 3 minutes at room temperature. Then, the blue reactant was transferred to a 96 well plate, and absorbance was measured at 750 nm. At this time, gallic acid was used as a standard material, and the result was expressed as mg gallic acid equivalent (GAE)/g extract.

<Results and evaluation>

Result 1 - Analysis of Total Polyphenol Content in Juniper Extract

The total polyphenol content of the juniper extract according to Examples 1 to 3 and Comparative Example 1 was confirmed through Experimental Example 1, and the results are shown in Table 1 below.

Juniper extract (C) according to Example 3 showed the highest total polyphenol content (78.6 mg GAE/g extract), and Juniper extract (B) according to Example 2 and Juniper extract according to Example 1 (A) showed the next highest total polyphenol content. In addition, compared with the polyphenol content shown in the juniper extract (D) according to Comparative Example 1, the polyphenol content was about 2 times higher in A, 2.7 times in B, and about 4.5 times higher in C.

Through this, it was confirmed that even if the same solvent, water, was used, the juniper extract containing a higher polyphenol content could be prepared according to the method for preparing the polyphenol-containing juniper extract of the present invention.

Claims (4)

preheating purified water to form pressurized hot water;
converting the pressurized hot water into a subcritical state to form subcritical water;
extracting the juniper tree with the subcritical water to form a juniper subcritical water extract; and
Method for producing a polyphenol-containing juniper extract comprising; filtering and deodorizing the juniper subcritical water extract.
The method of claim 1,
In the step of forming the subcritical water, the subcritical water is a method for producing a polyphenol-containing juniper extract using subcritical water, characterized in that it is formed in a temperature range of 300 to 350 ℃.
preheating purified water to form pressurized hot water;
converting the pressurized hot water into a supercritical state to form supercritical water;
hydrolyzing the juniper tree with the supercritical water to form a juniper hydrolyzate;
extracting the juniper hydrolyzate with subcritical water to form a juniper subcritical water extract; and
Method for producing a polyphenol-containing juniper extract comprising; filtering and deodorizing the juniper subcritical water extract.
4. The method of claim 3,
In the step of forming the supercritical water, the supercritical water is a method for producing a polyphenol-containing juniper extract, characterized in that it is formed in a temperature range of 374 to 425 ℃.

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