WO2023113391A1 - Method for efficiently extracting pinosylvin from pine trees - Google Patents

Abstract

The present invention relates to a method for efficiently extracting pinosylvin found in a trace amount from pine trees, the method comprising: a pine heartwood extraction step of selecting pine raw materials and extracting lipophilic heartwood; a powdering step of powdering the lipophilic heartwood into fine particles of 20 mesh or more; a primary extraction step of obtaining an extract from the powdered lipophilic heartwood; a primary concentration step of removing an organic solvent from the obtained primary extract; a secondary extraction step of obtaining an extract from the residue remaining after the extraction; a secondary concentration step of removing an organic solvent from the obtained secondary extract; and a mixing step of mixing the primary extract and the secondary extract.

Description

Method for Efficiently Extracting Pinosylvin from Pine Trees

The present invention relates to a method for extracting and obtaining pinosylvin from pine trees with high efficiency, which has high industrial utilization due to various physiological effects such as antioxidant, anticancer, cardioprotective, anti-inflammatory, and antibacterial.

Pinosylvin (3,5-dihydroxy- trans -stilbene) is a substance isolated and detected from plants damaged by biological effects such as insect invasion, bacterial and fungal infection, or physical effects such as UV and ozone. It is a natural secondary metabolite known to be biosynthesized mainly through the phenylalanine metabolic pathway. As a polyphenolic stilbene-based compound, it is also classified as phytoalexin, a defense substance produced by plants to prevent the growth of bacteria or fungi invading from the outside. According to what is known, pinosylbin is not produced in all plants but only in some plants, and it is known to be produced by Scottish fir, eucalyptus, spruce, grapes, peanuts, lilies, etc., as far as is known.

Among the stilbene compounds, resveratrol is the substance for which R&D has been most actively conducted. It is known to have anti-oxidant action. Resveratrol is mainly distributed in grape skins, grape seeds, peanuts, peanut sprouts, mulberries, and peony seeds. In particular, it is the cause of the French paradox phenomenon, which refers to the relatively low incidence of coronary heart disease among French people discovered in the late 1980s. It is a health functional indicator component that supports the claim that the cause is due to red wine.

It has been reported that resveratrol has a strong anticancer effect by blocking all three stages of carcinogenesis, initiation, promotion, and progression, as well as antiviral, neuroprotective, anti-inflammatory, antiaging, and life extension effects. Accordingly, botanists around the world have conducted diverse studies at the molecular and genetic level in relation to the biosynthetic mechanism and control of stilbene compounds. As a result, genes involved in the biosynthesis of stilbene compounds have been isolated from pine trees as well as grapes and peanuts, which are existing sources, and a lot of information on the expression and regulation of the genes has been accumulated.

In particular, as it was discovered that stilbene-based compounds present in plants make plants resistant to external stress factors, genes encoding factors involved in generating stilbene-based compounds among plants were introduced into specific crops to cause plant disease. There have been many cases of developing transgenic disease crops that are resistant to disease-causing factors such as fungi, bacteria, viruses, and pests that cause . Method for producing a transgenic plant with increased stilbene production and the resulting plant) corresponds to this case (Patent Document 1). In addition, in Registered Patent No. 10-0610875 (Title of Invention: Method for mass production of stilbene-based compounds derived from pine needles using environmental factors and their uses), various environmental factors that cause environmental stress, such as ultraviolet rays, ozone, aluminum chloride and yeast extract, are applied to pine needles. and disclosed a method for mass production of stilbene-based compounds derived from pine needles and their uses (Patent Document 2).

(Patent Document 1) Korean Registered Patent No. 10-1566692

(Patent Document 2) Korean Registered Patent No. 10-0610875

On the other hand, stilbene compounds including pinosylvin and resveratrol show various physiological activities such as antioxidant, anti-inflammatory and anti-cancer as well as antimicrobial effects such as antifungal, antibacterial and antiviral, so they can be used for various purposes in the food and drug industry. . Accordingly, by evaluating the physiological activity of stilbene-based compounds in various patent documents, anthelmintic effect (Patent Document 3, name of the invention: composition for enhanced anthelmintic activity and method for enhancing anthelmintic toxicity), bone disease treatment effect (Patent Document 4 , Title of the invention: extraction method of resveratrol and its derivatives for bone diseases isolated from peony seeds) or anti-inflammatory of pinosylvin (Patent Document 5, title of the invention: anti-inflammatory composition containing pinosylvin and rosmarinic acid), blood vessels Angiogenesis promotion (Patent Document 6, Title of Invention: Novel Angiogenesis Promotion Use of Pinosylvin), Prevention and Treatment of Cardiovascular Disease (Patent Document 7, Title of Invention: High Concentration Pinosylvin Composition for Prevention and Treatment of Cardiovascular Disease) ), etc. are mainly disclosed.

(Patent Document 3) Republic of Korea Patent Publication 10-1993-0011817

(Patent Document 4) Korean Registered Patent No. 10-0447607

(Patent Document 5) Korean Registered Patent No. 10-0795513

(Patent Document 6) Republic of Korea Patent Publication 10-2014-0103400

(Patent Document 7) Korean Registered Patent No. 10-1660626

Pinosylbin is known to be mainly isolated and detected from the leaves (pine needles) and wood of pine trees, but the amount of pinosylbin that can be obtained from pine trees is very small when it is actually separated and purified for industrial use. Therefore, according to the industrial utilization of pinosylbin, research and development are continuing to increase the content of pinosylvin, which is naturally present in a very small amount, or to obtain pinosylvin with high efficiency. In a preceding paper (Thesis 1, Title: Development of Ultrasonic Extraction Process for Production of High-Pinosylvin-Content Extract of Pine Needles), pinosylvin was extracted by applying an 80% ethanol aqueous solution and an ultrasonic extraction method to freeze-dried pine needles, resulting in a level of 0.2 mg per g. A trace amount of pinosylvin could be obtained. In addition, there has been a case of artificially increasing the production of pinosylvin by promoting the activity of stilbene biosynthetic enzymes using genetically modified microorganisms (Thesis 2, title: A novel process for obtaining pinosylvin using combinatorial bioengineering in Escherichia coli ).

(Thesis 1) Biosystems Engineering (2003) Vol. 28 No. 4: 325-334

(Paper 2) World J Microbiol Biotechnol (2016) 32(6): 102

The present invention has been made to solve the above-mentioned problems, and an object of the present invention is to select pine trees containing pinosylbin in order to obtain concentrated pinosylbin from pine trees, and among pine trees, areas containing high concentrations of pinosylbin. is extracted and extracted through several steps to maximize pinosylvin extraction efficiency. However, it is not limited to the technical problem as described above, and another technical problem may be derived from the following description.

A method for efficiently extracting pinosylbin from pine trees according to an embodiment of the present invention includes a pine heart material extraction step of selecting a pine raw material and extracting a lipophilic heart material; A powdering step of powdering the lipophilic core material into fine particles of 20 mesh or more; A first extraction step of obtaining an extract from the powdered lipophilic core material; and a secondary extraction step of obtaining an extract from the residue remaining after extraction.

In addition, the pine heartwood extraction step may include a selection step of selecting pine trees having a diameter of 50 cm or more; A drying step of drying the moisture content of the selected pine trees to 19% or less; A separation step of removing the outer skin and sapwood of the dried pine tree to separate a core material composed of a fibrous core material and a lipophilic core material; and a lipophilic core material extraction step of extracting the lipophilic core material by removing the fibrous core material from the core material.

In addition, the powdering step may include a powdering step of micronizing the particle size to 20 mesh or more in order to maximize the amount of pinosylbin eluted from the extracted lipophilic core material.

In addition, in the first extraction step, the powdered lipophilic core material is added to the organic solvent in an amount of 1 to 50% by weight and then heated at the boiling point of the solvent for 0.5 to 5 hours to ensure that pinosylvin is sufficiently eluted in the organic solvent It may include a first extraction step.

In addition, in the second extraction step, the residue generated in the first extraction step is added to an organic solvent in an amount of 1 to 50% by weight and then heated at the boiling point of the solvent for 0.5 to 5 hours so that pinosylvin is sufficiently eluted in the organic solvent. A secondary extraction step may be included.

In addition, the organic solvent, acetic acid, methanol, ethanol, n-propanol, isopropyl alcohol, n-butanol, formic acid, ammonia, chloroform, dichloromethane, ethyl acetate, acetone, dimethylformamide, acetonitrile, dimethyl sulfoxide A solution obtained by diluting at least one of side, nitromethane, propylene carbonate, 1,4-dioxane, diethyl ether, tetrahydrofuran, pentane, hexane, benzene, and toluene in distilled water at a ratio of 50 to 100% by volume may include

In addition, the pine extract containing pinosylbin according to another embodiment of the present invention is characterized in that it is prepared by a method for efficiently extracting pinosylbin from the above-described pine tree.

The method for extracting pinosylbin from pine trees according to the present invention selects pine trees containing a particularly large amount of pinosylbin, selects only lipophilic heartwood from wood parts, and uses it as an extract (solute) for pinosylvin extraction, as well as extracting pinosylbin. There is an effect of maximizing the extraction efficiency of pinosylvin from pine trees by removing pine parts having a low pinosylvin content using an organic solvent that is easy to use.

In particular, by pulverizing the extracted lipophilic core material into fine particles of 20 mesh or more, it is effective to maximize the surface area of the lipophilic core material exposed to the organic solvent so that pinosylvin is maximally eluted into the organic solvent. In addition, after obtaining the primary pine extract containing highly concentrated pinosylbin, a secondary pine residue extract additionally containing pinosylbin is obtained from the remaining residue, and mutually mixed to maximize the content of pinosylbin in the extract, thereby maximizing the pinosylbin per unit treatment. has the effect of maximizing the yield of

Through this, there was a problem in that the amount of pinosylvin extracted when trying to obtain pinosylvin from pine trees was extremely small. It has the effect of securing the economic feasibility of pinosylvin raw material with high industrial use due to various physiological activities by obtaining highly concentrated pinosylvin more efficiently by showing yield.

In addition, the method for extracting pinosylvin from pine trees according to the present invention and the pine extract containing pinosylbin prepared according to the method can show the commonly known physiological activity of pinosylvin, and have known uses (antioxidation, anti-aging, antibacterial). , antifungal, antiviral, anti-inflammatory, anticancer, promoting angiogenesis, prevention and treatment of cardiovascular diseases, etc.).

1 is a flowchart showing a method for efficiently extracting pinosylbin from pine trees according to an embodiment of the present invention.

Figure 2 is a flowchart showing the pine heartwood extraction step in the method of extracting pinosylbin from pine trees.

Figure 3 is a standard calibration curve for pinosylvin standard solution compared to high performance liquid chromatography (HPLC) peak range obtained for quantitative analysis of pinosylvin according to an embodiment of the present invention.

Figure 4 is a high-performance liquid chromatography chromatogram of pine heartwood extracts A, B, and C compared to pinosylbin 0.1 mg / ml standard solution according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. Embodiments described below relate to a method for extracting pinosylvin from pine trees with high efficiency compared to conventionally known methods.

The embodiments and drawings introduced below are provided as examples to sufficiently convey the spirit of the present invention to those skilled in the art. In addition, unless otherwise defined, the technical terms and scientific terms used in the present invention have meanings commonly understood by those of ordinary skill in the art to which this invention belongs, and in the following description and accompanying drawings, the present invention Descriptions of well-known functions and configurations that may unnecessarily obscure the gist of will be omitted.

Hereinafter, the technical idea of the present invention will be described in more detail using the accompanying drawings. Since the accompanying drawings are only examples shown to explain the technical idea of the present invention in more detail, the technical idea of the present invention is not limited to the form of the accompanying drawings.

1 is a flowchart showing a method for efficiently extracting pinosylbin from pine trees according to an embodiment of the present invention, and FIG. 2 is a flowchart showing the pine heartwood extraction step in the method for extracting pinosylbin from pine trees in detail. am.

Referring to FIG. 1, in the method of extracting pinosylbin from pine trees according to an embodiment of the present invention, a pine heart material extraction step (S100) of selecting a pine raw material and extracting a heart material, and a powdering step of pulverizing the extracted pine heart material (S200), a first extraction step of extracting pinosylvin from powdered pine heartwood with an organic solvent (S300), and a second extraction step (S400) of extracting pinosylvin from the residue generated in the first extraction step with an organic solvent. can include Below, each step is described in detail.

The pine heartwood extraction step (S100) is a step of selecting pine raw materials and drying them to extract the lipophilic core material therein. In detail, the pine heartwood extraction step (S100) may include a log selection step (S110), a drying step (S120), a heartwood separation step (S130), and an oleophilic heartwood extraction step (S140), which can be referred to in FIG. there is.

The raw wood sorting step (S110) is a step of sorting hardwood, that is, pine, for extracting the heartwood. At this stage, pine trees with a diameter of 50 cm or more can be selected. When the diameter of a pine tree is less than 50 cm, the content of polyphenols extracted from the heartwood or the level of antioxidant power is low, or the amount of the extract eluted with an organic solvent is less than the set standard in many cases, and in this case, the present invention is difficult to achieve the intended purpose. Since this is difficult, you can select pine trees with a diameter of more than 50 cm and proceed to the next step.

The next step, the drying step (S120), is a step of drying the selected pine logs. In detail, after logging the selected pine logs, the logs can be dried until the moisture content of the logs is 25% or less, preferably 19% or less. The drying method for drying such raw wood can mainly utilize natural drying, and drying can be performed for more than 25,000 hours, approximately 3 years or more in an environment of -10 to 40 ° C and a humidity of 45 to 85%. .

Accordingly, the moisture content of pine wood can be dried to 19% or less, and in the case of such pine wood, when a blow is applied to the wood with a wooden hammer, moisture is condensed inside and a clear bell sound can be heard, based on this, the moisture content is 19 % or less. However, this is one method, and the moisture content inside the tree can be measured by inserting the humidity meter into the tree and waiting for the humidity measurement set time.

The core material separation step (S130) is a step of separating the core material by removing the outer skin and sapwood of the pine log dried in the drying step (S120). In detail, a pine log can be divided into a core formed in the inner center, a sapwood surrounding the core, and an outer skin surrounding the sapwood and forming an exterior. This is because most of the antioxidants and polyphenols are contained in the heartwood among the constituents of raw wood, so only these parts are separated separately to maximize the scent of pinosylvin extracted from pine trees.

The lipophilic core material extraction step (S140) is a step of obtaining a lipophilic core material by separating the lipophilic core material containing the polyphenol content and fragrance component from the core material and the fibrous core material on which lignin or the like is deposited. Among the core materials, the lipophilic core material with a high content of aromatic components and polyphenols is a characteristic part for constituting the effect of the present invention, and in the case of a fibrous core material, the extraction efficiency of pinosylvin is greatly reduced due to the high content of lignin, which is a simple fiber. am.

The powdering step (S200) of the method of extracting pinosylbin from the pine tree of FIG. 1 is a step of extracting the pine core material and then pulverizing the extracted pine core material. In the powdering step (S200), the particle size of the extracted lipophilic core material can be powdered to a level of fine particles of 20 mesh or more. This allows pinosylvin, the desired component, to be eluted to the maximum extent from the lipophilic core material, thereby increasing the extraction amount of pinosylvin as well as extracting the desired component within a short period of time.

In detail, in the powdering step (S200), the separated lipophilic core material is brought into contact with the impeller rotating at a set speed, and the lipophilic core material powder atomized to 20 mesh or more generated by the frictional force between the impeller and the lipophilic core material is obtained. It may be made in the form of, but it is not limited to this, and equipment capable of atomizing the lipophilic core material into particles of 20 mesh or more can be utilized.

In detail, in the first extraction step (S300), the pine heartwood powder obtained in the powdering step (S200) is mixed with an organic solvent at a ratio of 1 to 50% by weight and heated at the boiling point for 0.5 to 5 hours to dissolve pinosylvin in the organic solvent. This step is to ensure sufficient elution. This is because the amount of pinosylvin eluted in the organic solvent may be extremely small when the weight % of the pine heartwood powder is lower than this, and conversely, when the amount exceeds 50% by weight, pinosylbin is released from the organic solvent in which the organic components of the pine tree are eluted. This is because it is difficult to separate the pine extract containing it.

In detail, in the first concentration step (S400), a vacuum concentration method may be applied to obtain a pure extract from which the organic solvent is completely removed, and finally, it may be exposed to a vacuum state. This is because, when the organic solvent remains in the extract, the content of pinosylbin in the total weight is low, and the desired effect may be reduced by treatment of the extract.

In detail, in the second extraction step (S500), the pine heartwood powder residue obtained in the first extraction step (S300) is mixed with an organic solvent in an amount of 1 to 50% by weight and heated at the boiling point for 0.5 to 5 hours to dissolve the organic solvent. This step is to ensure that silvin is sufficiently eluted. This is because the amount of pinosylvin eluted in the organic solvent may be extremely small when the weight% of the pine heartwood powder residue is lower than this, and conversely, when the amount exceeds 50% by weight, the pinosylvin from the organic solvent in which the organic components of the pine tree are eluted. This is because it is difficult to isolate the pine extract containing .

In detail, in the secondary concentration step (S600), a vacuum concentration method may be applied to obtain a pure extract from which the organic solvent is completely removed, and finally, it may be exposed to a vacuum state. This is because, when the organic solvent remains in the extract, the content of pinosylbin in the total weight is low, and the desired effect may be reduced by treatment of the extract.

In addition, the mixing step (S700) is a step of mutually mixing the pine extract prepared through the first concentration step (S400) and the second concentration step (S600), and acetic acid to facilitate the acquisition of pinosylvin during the mixing process , methanol, ethanol, n-propanol, isopropyl alcohol, n-butanol, formic acid, ammonia, chloroform, dichloromethane, ethyl acetate, acetone, dimethylformamide, acetonitrile, dimethyl sulfoxide, nitromethane, propylene Organic solvents such as carbonate, 1,4-dioxane, diethyl ether, tetrahydrofuran, pentane, hexane, benzene, toluene, etc. may be added, but are not limited thereto, and are generally referred to as organic solvents. Note that all are included.

Hereinafter, differences in pinosylvin extraction efficiency that may occur depending on the age and part of pine trees according to an embodiment of the present invention will be described in detail. The following experiment was conducted to confirm the difference in pinosylvin extraction efficiency according to the age and part of pine trees used as raw materials. The experimental name for the experiment is defined as follows.

Experiment 1. Comparison of pinosylvin extraction efficiency according to the age of pine trees

Experiment 2. Comparison of pinosylvin extraction efficiency according to the difference in pine tree parts

1. Experiment conditions

(1) Analysis target sample

The samples to be analyzed were pine trees aged about 55 and 97 years with a diameter at the chest height of less than 50 cm and heartwood of pine trees with a diameter at the chest height of 61 to 78 cm at the age of 180 to 250 years corresponding to an embodiment of the present invention. To confirm pinosylvin extraction efficiency, leaves, branches, and sapwood parts were used. Pinosylvin standard material used for quantitative analysis of pinosylbin was 98% or higher purity pinosylvin (product number: PHL805160) purchased from Sigma aldrich.

(2) Experimental conditions and experimental methods

The experiment utilized a 70% methanol extraction method and proceeded in the following order.

go. Sample collection and powderization (size of 20 mesh or more)

me. Add 100 mL of 70% methanol to 10 g of powdered sample in a round bottom flask and cool for more than 30 minutes.

all. After heating the mixture of the sample and 70% methanol in a heating extractor (65℃) for 3 hours, cool it quickly and transfer it to a separatory funnel

la. This concentrate was concentrated and dried under nitrogen gas on a water bath at 40°C or less.

mind. Filter this with a 0.45 ㎛ membrane filter and use it as a test solution

bar. Put the test solution in a vial and measure the result by high-performance liquid chromatography

In addition, the test equipment for instrumental analysis used the test model below, and the instrumental analysis conditions were set as follows.

- Column: YMC ODS-A column (4.6 × 150 mm, 5 μm)

- Instrument: Agilent 1200 HPLC system

- Instrumental analysis conditions are shown in Table 1

Instrument	Agilent 1200 HPLC system
Column	YMC ODS-A column
Injection vol.	10 µL
Column temp. (℃)	30℃
Flow rate	System 1 (HPLC): 0.7 mL/min
UV wavelength	System 1 (HPLC): 210, 254, 280, 330 nm
Mobile phase	Solvent A: 0.02 TFA in Acetonitrile Solvent B: 0.02% TFA in water

For quantitative analysis of pinosylvin, standard solutions prepared by diluting pinosylvin at 6 concentrations (0.001, 0.005, 0.01, 0.05, 0.1 and 0.5 mg/mL) were prepared and compared to peak ranges of high-performance liquid chromatography for pinosylvin standard solutions. A calibration curve was obtained, and the standard calibration curve is shown in Table 2 and FIG. 3 below.

Pinosylvin conc. (mg/mL)	Peak area
0.001	93.28
0.005	337.09
0.01	740.58
0.05	3647.93
0.1	7771.22
0.5	20112.80

3. Experimental results

(1) Experimental results under Experiment 1 conditions

Referring to Table 3 and FIG. 4 below, the experimental results of Experiment 1 conditions for comparing pinosylvin extraction efficiency according to the age difference of pine trees can be confirmed.

pine tree			pinosylvin
classification	age (years)	bust diameter (cm)	content in the extract (mg/g)	content compared to sample (mg/g)
Comparative Example 1	55	25	1.55	0.06
Comparative Example 2	97	43	6.20	0.23
Example 1	180	61	12.37	0.41
Example 2	250	78	9.57	0.55
Example 3	190	68	13.24	0.40

According to one embodiment of the present invention, the content of pinosylvin extracted from the heartwood of pine trees with a breast height of 50 cm or more is 0.40-0.55, whereas the content of pinosylvin extracted from the heartwood of pine trees with a breast height of 50 cm or less is 0.06-0.23 mg/g. As shown, it was confirmed that the extraction yield of pinosylvin according to an embodiment of the present invention was remarkably high.

(2) Experimental results under Experiment 2 conditions

Referring to Table 4 below, it can be seen the experimental results of Experiment 2 conditions for comparing the extraction efficiency of ㄸ and other pinosylbin in the difference in the parts of pine trees.

pine tree				pinosylvin
classification	age (years)	bust diameter (cm)	part	content in the extract (mg/g)	content compared to sample (mg/g)
Comparative Example 1	55	25	heartwood	1.55	0.06
Comparative Example 2	97	43	heartwood	6.15	0.23
Comparative Example 3	180	61	leaf	1.15	0.04
Comparative Example 4			egg plant	8.67	0.32
Comparative Example 5			sapwood	7.65	0.28
Example 1			heartwood	12.37	0.41
Comparative Example 6	250	78	leaf	1.01	0.04
Comparative Example 7			egg plant	7.55	0.28
Comparative Example 8			sapwood	6.43	0.24
Example 2			heartwood	9.57	0.55
Comparative Example 9	190	68	leaf	1.11	0.04
Comparative Example 10			egg plant	8.53	0.32
Comparative Example 11			sapwood	7.77	0.29
Example 3			heartwood	13.24	0.40

According to an embodiment of the present invention, the content of pinosylvin extracted from the heartwood of a pine tree having a chest height of 50 cm or more is 0.40-0.55, while the content of pinosylvin extracted from other parts such as leaves, branches, and sapwood of the same pine tree is 0.04-0.32 It was confirmed that the amount of pinosylbin extracted from the lipophilic heartwood of pine trees with a breast height of 50 cm or more in diameter was remarkably high. In particular, pine trees containing a lot of pine trees are selected, and only lipophilic heartwood is selected from wood parts to be used as an extract (solute) for pinosylbin extraction. Also, pine parts with low pinosylbin content are selected by using an organic solvent that is easy to extract pinosylvin. By removing it, there is an effect of maximizing the extraction efficiency of pinosylvin from pine trees.

In particular, by pulverizing the extracted lipophilic core material into fine particles of 20 mesh or more, it is effective to maximize the surface area of the lipophilic core material exposed to the organic solvent so that pinosylvin is maximally eluted into the organic solvent. In addition, after obtaining the primary pine extract containing highly concentrated pinosylbin, a secondary pine residue extract additionally containing pinosylbin is obtained from the remaining residue, and mutually mixed to maximize the content of pinosylbin in the extract, thereby maximizing the pinosylbin per unit treatment. has the effect of maximizing the yield of

Through this, there was a problem in that the amount of pinosylvin extracted when trying to obtain pinosylvin from pine trees was extremely small. It has the effect of securing the economic feasibility of pinosylvin raw material with high industrial use due to various physiological activities by obtaining highly concentrated pinosylvin more efficiently by showing yield.

In addition, the method for extracting pinosylvin from pine trees according to the present invention and the pine extract containing pinosylbin prepared according to the method can show the commonly known physiological activity of pinosylvin, and have known uses (antioxidation, anti-aging, antibacterial). , antifungal, antiviral, anti-inflammatory, anticancer, promoting angiogenesis, prevention and treatment of cardiovascular diseases, etc.).

So far, the present invention has been looked at with respect to preferred embodiments. Those skilled in the art to which the present invention pertains will be able to understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an illustrative rather than a limiting point of view. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the equivalent scope will be construed as being included in the present invention.

Claims (10)

1. Pine heartwood extraction step of selecting pine raw materials and extracting lipophilic heartwood;

   A powdering step of powdering the lipophilic core material into fine particles of 20 mesh or more;

   A first extraction step of obtaining an extract from the powdered lipophilic core material;

   A first concentration step of removing organic solvents from the extracted first extract;

   A secondary extraction step of obtaining an extract from the residue remaining after extraction;

   Secondary concentration step of removing the organic solvent from the extracted secondary extract; and

   Method for efficiently extracting pinosylbin from pine, characterized in that it comprises a mixing step of mixing the primary extract and the secondary extract.
2. According to claim 1,

   The pine raw material is a species belonging to the genus Pinus spp. in the botanical classification Pinus spp. Common names pine or pine ( Pinus densiflora ), black pine or sea pine ( Pinus thunbergia ), pine tree ( Pinus rigida ), pine tree ( Pinus koraiensis ) ), a method for efficiently extracting pinosylbin from a pine tree, characterized in that at least one of a pine tree ( Pinus parviflora ), and a snow pine tree ( Pinus pumila) .
3. According to claim 1,

   The pine heartwood extraction step,

   A screening step of selecting pine trees with a diameter of 50 cm or more;

   A drying step of drying the moisture content of the selected pine trees to 19% or less;

   A separation step of removing the outer skin and sapwood of the dried pine tree to separate a core material composed of a fibrous core material and a lipophilic core material; and

   A method for efficiently extracting pinosylbin from pine, characterized in that it comprises a lipophilic heart material extraction step of extracting the lipophilic heart material by removing the fibrous heart material from the heart material.
4. According to claim 1,

   The powdering step is a method for efficiently extracting pinosylbin from pine, characterized in that the size of the particles is micronized to 20 mesh or more to maximize the amount of pinosylbin eluted from the extracted lipophilic core material.
5. According to claim 1,

   In the first extraction step, the powdered lipophilic core material is added to the organic solvent in an amount of 1 to 50% by weight and then heated at the boiling point of the solvent for 0.5 to 5 hours so that pinosylvin is sufficiently eluted in the organic solvent. A method for extracting pinosylbin from pine trees.
6. According to claim 1,

   The primary concentration step is a method for extracting pinosylbin from pine, characterized in that to obtain a pure extract from which the organic solvent is completely removed in a vacuum state after concentrating the organic solvent including the extract in the primary extraction step under reduced pressure.
7. According to claim 1,

   In the second extraction step, the residue generated in the first extraction step is added to an organic solvent in an amount of 1 to 50% by weight and then heated at the boiling point of the solvent for 0.5 to 5 hours so that pinosylvin is sufficiently eluted in the organic solvent. A method for extracting pinosylbin from pine trees.
8. According to claim 1,

   The secondary concentration step is a method for extracting pinosylbin from pine, characterized in that to obtain a pure extract from which the organic solvent is completely removed in a vacuum state after concentrating the organic solvent including the extract in the secondary extraction step under reduced pressure.
9. According to claims 5 to 8,

   The organic solvent is acetic acid, methanol, ethanol, n-propanol, isopropyl alcohol, n-butanol, formic acid, ammonia, chloroform, dichloromethane, ethyl acetate, acetone, dimethylformamide, acetonitrile, dimethyl sulfoxide, A solution obtained by diluting at least one of nitromethane, propylene carbonate, 1,4-dioxane, diethyl ether, tetrahydrofuran, pentane, hexane, benzene, and toluene in distilled water at a ratio of 50 to 100% by volume A method for extracting pinosylbin from pine trees.
10. A method for extracting pinosylvin from pine, characterized in that the extraction by the method of any one of claims 1 to 9.

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