KR100659945B1 - Extraction method for antioxidants from pine trees using natural solvents and antioxidant extracts thereby - Google Patents

Abstract

The present invention relates to a method for extracting antioxidants derived from pine bark using a natural solvent as an extracting medium, and to extracting the antioxidant from the bark of pine, using an alcoholic solution as a natural solvent from pine bark; Separating the non-polar substance from the primary extract using a natural alcohol solution and / or natural animal and vegetable oil for the primary extract, and the natural antioxidants with excellent antioxidant activity from pine bark It is an invention for providing economically.

Pine Bark, Antioxidants, Natural Solvents, Extraction, Separation

Description

Extraction method for antioxidants from pine trees using natural solvents and antioxidant extracts-1

The present invention relates to a method for extracting an antioxidant substance derived from pine bark using a natural solvent as an extraction medium and an antioxidant extract thereby.

Recently, as the elderly population is rapidly increasing and the main cause of various adult diseases is found to be free radicals, research and interest on natural antioxidants have been greatly increased.

Free radicals are oxygen that has free radicals and have very unstable chemical properties and show high reactivity to the surrounding materials, thereby oxidatively damaging cellular proteins, lipid molecules, and genetic material DNA. It is known to be very fatal to function and life.

Antioxidants are substances that have scavenging activity of these free radicals produced in the human body and prevent and improve aging and various diseases (arteriosclerosis, hypertension, cerebral hemorrhage, diabetes, heart disease, etc.) caused by oxidative stress accumulated in the human body. As a result, many studies have been conducted, and there is an urgent need for the development of powerful natural antioxidants with therapeutic functions for various diseases.

Antioxidants to remove the active oxygen can be largely divided into synthetic antioxidants and natural antioxidants.

Synthetic antioxidants such as BHA (Butylated hydroxyanisole, Butyrated hydoxyanisole), BHT (Butylated hydroxytoluene, Butyrated hydoxytoluene) are widely used, but recent studies have shown that they are toxic to synthetic antioxidants. There is a need for the development of natural antioxidants that are safe and have excellent antioxidant activity.

It is known that pine bark contains components of bioflavonoids, a natural antioxidant that strongly removes free radicals, and there have been many patent applications for extracting antioxidants from pines.

According to US Pat. No. 4,698,360, a synthetic solvent (ethyl acetate, chloroform) is repeatedly used to extract and separate antioxidants from pine bark, and inorganic salts (salt, ammonium sulfate, sodium sulfate) are used. Therefore, the manufacturing process is very complicated and the economic efficiency is very low due to the very low extraction yield (about 0.21% of the bark), and also due to the synthetic organic solvent remaining in the antioxidant extract, its safety is a problem for use as food.

According to Korean Patent No. 10-0212974, workability is very high due to the use of highly volatile synthetic organic solvents such as methanol, diethyl ether, ethyl acetate, heptane, etc. to extract and separate antioxidants from pine bark. Poor and also difficult to use as a food due to the possibility of the residual organic solvent in the antioxidant extract.

In addition, Korean Patent No. 10-0429406 shows a method for preparing an antioxidant derived from pine needles. In this case, synthetic organic solvents such as methanol and ethyl acetate are used, and Korean Patent Publication No. 199-0005694 is an antioxidant from green tea leaves. The manufacturing method of the present invention is shown. In this case, the extract is extracted by adding calcium carbonate to the water, and then the antioxidant is adsorbed using an adsorptive resin, followed by the use of synthetic organic solvents such as ethyl acetate, methanol and acetone. Is separating.

Furthermore, Korean Patent No. 10-0341797, which shows a method for separating and purifying natural antioxidants from safflower seeds, also uses a synthetic organic solvent such as methanol, nucleic acid, ethyl acetate, and chloroform to separate the antioxidants. Korean Patent No. 10-0109461, which shows the separation method of antioxidant components, also uses synthetic organic solvents such as methanol, ethyl acetate, and chloroform, and Korean Patent No. 10-0395464, which shows a method for extracting catechins from green tea leaves. In addition, synthetic organic solvents such as dichloromethane, ethyl acetate, acetone, etc. are used. In Korea Patent No. 10-0439012, which shows an extraction method of antioxidants from the liana, synthetic organic solvents such as methanol and ethyl acetate are also used. Doing.

As described above, according to the conventional methods, a large amount of synthetic organic solvents are used in the manufacturing process of the antioxidant, and thus, safety and legal problems are raised in using the extract as a food raw material, and volatile organic solvents must be handled. As a result, the working conditions are very demanding, and various problems occur in the quality of the antioxidant and the manufacturing process such as generating a large amount of environmental pollutants that pollute the air and water.

In particular, according to the conventional extraction method, the extraction and separation of antioxidants from pine bark consists of several complex extraction, concentration and separation processes, resulting in a very low extraction yield of antioxidants. In addition, there is a problem that the production cost is high and the production cost is very high. Also, since a large amount of various volatile synthetic organic solvents that are toxic to the human body are used during the manufacturing process of the antioxidant, work safety is low, and air pollution and water pollution can be caused. There is a serious problem that the antioxidant extract can not be used as food due to the possibility of remaining of the synthetic organic solvent in the produced antioxidant extract.

In addition, in the conventional method, since various salts are used during the extraction and separation process, that is, salt, ammonium sulfate, sodium sulfate, calcium carbonate, etc., an additional step of completely separating the salts from the antioxidant material is required, resulting in very low workability and productivity. Environmental problems such as water pollution caused by the problem will occur.

The present invention provides a high-efficiency extraction process of antioxidants from pine bark and extracts by using only fermentation alcohol solution, which is a low-volatility and excellent anti-oxidant extraction ability, to solve the problems posed by the conventional method. In order to efficiently remove non-polar substances such as rosin and to separate and concentrate antioxidants, alcoholic beverages and / or natural animal and vegetable oils are used to simplify the manufacturing process and minimize the generation of environmental pollutants to reduce the environmental treatment costs. It is to provide an extraction method that can be saved. In addition, the anti-oxidant extract derived from pine bark obtained by the extraction process according to the present invention is not only excellent in antioxidant activity but also very high in extraction yield, and the extraction method according to the present invention is an antioxidant substance having a high price competitiveness from pine bark. Can be produced in large quantities.

The present invention primarily extracts antioxidants from the bark using a fermented alcohol solution, which is a natural solvent, in order to extract natural antioxidants from the bark of pine trees, rosin and the like contained in the extract of bark using alcohol or natural animal and vegetable oils. It is an object of the present invention to provide a method for extracting natural antioxidants, characterized in that the removal of non-polar substances efficiently.

In addition, the present invention specifies conditions for ethanol concentration, amount of use, extraction temperature, etc., in order to increase the antioxidant activity and extraction efficiency of the antioxidant extract, and in order to efficiently remove the non-polar substances contained in the extract ethanol solution or animal and vegetable oil It is another object to provide an efficient method of extracting natural antioxidants by specifying the conditions for the amount of solvent used, extraction temperature, filtration and concentration method using.

Furthermore, the present invention is to provide a natural antioxidant extract obtained by the natural antioxidant extracting method, in particular, extracts having utility as raw materials and additives, such as food, cosmetics, pharmaceuticals and feed.

According to the present invention, by using only a chemically relatively stable natural solvent to increase the safety and workability, and provides an environment-friendly process, and through the simplified manufacturing process step to increase the extraction yield and antioxidant activity of antioxidants to maximize economical Raised.

The present invention relates to a method for extracting pine bark-derived antioxidants, the bark pretreatment process, bark grinding process, alcohol solution extraction process, extract concentration or drying process, non-polar substances by alcohol and / or animal and vegetable oils. It will have an extraction process and an antioxidant drying process.

In the present invention, 'pine' is defined as a plant belonging to the pine family, specifically Red pine (Pinus densiflora Sieb & Zucc.), Sea pine (Pinus thumbergii Parlatore), Pinus densiflora Sieb. & Zucc. erecta Uyeki, Pinus sylvestris L., Pine Pine (Pinus pinea, Cupressus Sempervirens), Casuarina equisetifolia JR & G. Forst., Casuarina litorea L. Casuarinaceae or Himalayan Pine (Abies pindrow) May be exemplified.

In addition, the bark refers to the bark areas that can be obtained from trunk and branch parts, except for the leaves or roots of pineaceae plants, and the pretreated bark is preferably 5 to 100 to improve solvent extraction. Can be milled into a mesh.

In addition, the term "alcoholic solution" or "fermented alcoholic solution" referred to in the whole process of the present invention is defined as a fluid liquid of the ethanol main component fermented with a starch-containing material or a sugar-containing material. .

The concentration of ethanol in the alcoholic liquor used in the bark extraction process is 1.0-95%, preferably 30-95%, and the ethanol concentration in the alcoholic liquor used in the extraction process of the nonpolar material is 10-95% preferably. 40 ~ 95% is applied, and the natural animal and vegetable oils used in the extraction process of non-polar substances include soybean oil (soybean oil), corn oil (fertilized oil), rapeseed oil, rice bran oil, sesame oil, perilla oil, safflower oil, sunflower oil, cottonseed oil, Peanut oil, olive oil, palm oil, palm oil, mixed cooking oil, refined common oil, shortening, chili seed oil, tallow or lard may be exemplified.

Hereinafter, the method of the present invention will be described in detail for each process. However, these do not limit the technical scope of the present invention.

1. Bark pretreatment process: Pine bark is used for the grinding process by removing the foreign matter adhering to the outside and the black weathered bark part of the outermost layer and separating only the bark of the red part.

2. Bark grinding process: The separated bark is made into a particle or powder state using a crusher and a crusher, and then used for the extraction process. Preferably it is ground to 5 to 100 mesh.

3. Alcohol solution extraction process: First extract the antioxidant from the ground bark powder using fermented alcohol solution as a natural solvent. The content of ethanol contained in the alcohol solution is used 1.0 to 95%, preferably the content of ethanol 30 to 95%. The ratio of the bark: alcohol solution is used by mixing in a weight ratio of 1: 3 to 1:50, preferably in a weight ratio of 1: 4 to 1:20 to extract the antioxidant. Extraction temperature is -20 ~ 90 ° C, preferably 10 ~ 70 ° C and extraction time is 5 minutes to 3 days, preferably 30 minutes to 24 hours for stationary or intermittent or continuous mixing Extracts antioxidants.

4. Extraction Concentration or Drying Process: After the primary extraction is completed, the extract and the bark are separated using known methods such as filtration or centrifugation to separate the antioxidant extract from the bark. In the filtration process, diatomaceous earth, white clay, bentonite, acidic clay, talc, perlite, activated carbon, etc. may be used as the filtration aid. The alcohol solution in the bark filter foil is mixed 1 to 20 times, preferably 3 to 15 times compared to the filter foil, and the antioxidant remaining in the bark filter foil is further extracted once or twice. The filtered extract is mixed and concentrated by removing the ethanol and water by a known vacuum concentration method so that the solid content of the extract concentrate is 1.0-30%, preferably 3.0-15%. The extract concentrate is dried as it is or by a known drying method, for example, lyophilization, reduced pressure drying, hot air drying or spray drying, and then used to separate the non-polar material.

5. Separation process of non-polar substances by alcohol solution and / or natural animal and vegetable oil: To separate non-polar substances such as rosin from extract concentrate or dried extract powder, one of two processes can be selectively applied as follows. Concentrate the antioxidants.

(1) When using alcohol solution as a separation medium: Alcohol solution for separating non-polar substances is 10 to 95% of ethanol, preferably 40 to 95%. The concentration of the extract concentrate or extract powder: alcohol solution is mixed in a ratio of 1: 1 to 1:30 by weight, preferably 1: 3 to 1:15, and then the non-polar substances are separated to concentrate the antioxidant. Extraction temperature is extracted at -20 ~ 90 ° C, preferably 0 ~ 70 ° C and extraction time is 5 minutes to 24 hours, preferably 30 minutes to 10 hours for stationary or intermittent or continuous mixing To separate the non-polar material. Alcohol solutions containing antioxidants and nonpolar substances can be separated by known filtration or centrifugation methods.

(2) When using natural animal and vegetable oils as separate media: natural animal and vegetable oils, that is, soybean oil (soybean oil), corn oil (fertilized oil), rapeseed oil, rice bran oil, sesame oil, perilla oil, safflower oil, sunflower oil, cottonseed oil, peanut oil, Olive oil, palm oil, palm oil, mixed edible oil, refined common oil, shortening, red pepper seed oil, tallow or pork fat alone or a mixture of two or more kinds thereof may be used. The ratio of the extract concentrate or extract powder: natural oil is 1: 0.5 to 1:20 by weight ratio, preferably 1: 1 to 1:10 by mixing the ratio of the non-polar substances to separate the oil layer to concentrate the antioxidant. Separation temperature of non-polar material is extracted at 0 ~ 120 ° C, preferably 20 ~ 80 ° C and extraction time is 30 minutes ~ 24 hours, preferably 1.0 hours ~ 10 hours for stationary or intermittent mixing or continuous The nonpolar material is separated by mixing. The concentrated antioxidant extract and natural oils containing nonpolar substances can be separated by known filtration or centrifugation.

Optionally, when the non-polar substance is to be separated using the extract powder, the non-polar substance may be extracted using a natural oil after adding 5.0 to 95% of the alcoholic beverage at a weight ratio of 1.0 to 20 times that of the extract powder. At this time, the separation temperature of the non-polar material is extracted at -20 ~ 90 ° C, preferably 10 ~ 80 ° C and the extraction time is 30 minutes to 24 hours, preferably 1.0 hours to 10 hours The nonpolar material is separated by continuous mixing.

6. Antioxidant Drying Process: Antioxidant concentrated extracts from which nonpolar substances have been removed are prepared in the form of concentrated liquid or solid powder by known concentration, drying and grinding processes.

<Example 1>

1) Extraction and filtration process: After removing the outermost layer from the bark obtained from the 30-year-old red pine (Pinus densiflora Sieb & Zucc.) Growing in Samcheok, Gangwon-do, Korea, a bark powder (30 mesh) was obtained using a crusher and a grinder. 1.0Kg bark powder was mixed with 8.0Kg of natural alcohol solution containing 60% ethanol, and then extracted in a shaking constant temperature water bath heated at 50 ° C for 3 hours and filtered. After adding 4.0Kg of alcohol solution consisting of 60% ethanol and 40% water to the bark filter cake, the mixture was extracted for 1 hour in a shaking constant temperature bath heated at 50 ° C and filtered.

2) Concentration and Drying Process: After extracting the filtrates obtained in step 1), ethanol and water were removed using a vacuum concentrator to obtain 1,230 g of an antioxidant primary extract concentrate. The extract concentrate was spray dried to obtain 95 g of antioxidant powder.

3) Separation of nonpolar substances and drying of antioxidants: After mixing 100g of alcohol with 30g of antioxidant powder obtained in 2) process, the nonpolar substances were separated and filtered in a 30 ° C shaking constant temperature water bath for 3 hours. Filter foil containing antioxidants was dried at 50 ° C for 3 hours and then ground to give 14.2 g of reddish brown antioxidant powder.

 <Example 2>

In the separation and nonoxidation of the nonpolar substance of Example 1, the nonpolar substance was extracted and removed in a 30 ° C shaking constant temperature water bath for 5 hours using 70 g of alcohol and 100 g of soybean oil instead of 100 g of alcohol, followed by centrifugation. Soybean oil layer and alcohol layer were separated. Solid non-polar antioxidants were dried at 50 ° C for 3 hours and then ground to give 13.2 g of reddish brown antioxidant powder.

<Example 3>

Antioxidants were prepared in the same manner except for using alcohol containing 95% ethanol in the extraction and filtration process of Example 1. Finally, reddish brown antioxidant powder obtained 12.3 g.

 <Example 4>

Antioxidants were prepared in the same manner except for using a ethanol solution containing 30% ethanol in the extraction and filtration process of Example 1. Finally, reddish brown antioxidant powder obtained 13.5 g.

Example 5

In the non-polar material separation and antioxidant drying process of Example 3, non-polar material was extracted and removed for 5 hours in a 30 ° C shaking constant temperature water bath by using 70 g of alcohol and 100 g of soybean oil instead of 100 g of alcohol, followed by centrifugation. One soybean oil layer and a ethanol solution layer were removed. Solid non-polar antioxidants were dried at 50 ° C for 3 hours and then ground to give 12.1 g of reddish brown antioxidant powder.

<Example 6>

In the nonpolar substance separation and antioxidant drying process of Example 4, the nonpolar substance was extracted and removed in a 30 ° C shaking constant temperature water bath for 5 hours using 80g of alcohol and 120g of soybean oil instead of 100g of alcohol. The soybean oil layer and the alcohol solution layer were removed. Solid non-polar antioxidants were dried at 50 ° C for 3 hours and then pulverized to give 11.3 g of reddish brown antioxidant powder.

<Example 7>

In the nonpolar material separation and antioxidant drying process of Example 1, the nonpolar material was extracted for 5 hours in a 5 ° C shaking constant temperature water bath and then filtered. The filtered foil containing concentrated antioxidants was dried at 50 ° C. for 3 hours and then ground to give 15.7 g of reddish brown antioxidant powder.

<Example 8>

In the nonpolar material separation and antioxidant drying process of Example 3, the nonpolar material was extracted for 5 hours in a 5 ° C shaking constant temperature water bath, and then filtered. Filter foil containing concentrated antioxidants was dried at 50 ° C for 3 hours and then ground to give 13.5 g of reddish brown antioxidant powder.

Comparative Example 1

Pine bark extract was prepared by the same method as that disclosed in US Pat. No. 4,698,360 as follows. The bark used for extraction was 100g of bark powder (30 mesh) isolated from red pine native to Samcheok, Gangwon-do. 500ml of distilled water was mixed with bark and heated at 100 ° C. for 2 hours to extract antioxidants and filtered. The filtrate was cooled to 20 ° C and salt was added until saturated, and the resulting precipitate was filtered off. Antioxidant was separated from the aqueous layer by mixing 100 ml of ethyl acetate in the extract filtrate. The separation process using ethyl acetate was repeated two more times, and then all of the ethyl acetate layers were combined and dehydrated by adding 5.0 g of sodium sulfate thereto. The dehydrated ethyl acetate layer was removed 80% of the ethyl acetate by vacuum concentration. 200 ml of chloroform was added to the concentrated ethyl acetate layer to obtain a precipitate, and the precipitate was separated by filtration. The precipitate was finally washed by adding 50 ml of chloroform and the precipitate was dried at 40 ° C. Finally, 0.21 g of antioxidant powder was obtained.

Comparative Example 2

Pine bark extract was prepared by the same method as the method disclosed in Korean Patent No. 10-0212974 as follows. The bark used for extraction was 100g of bark powder (30 mesh) isolated from red pine native to Samcheok, Gangwon-do. A mixture of 1.0 liter methanol and water (methanol: water = 3: 1 v / v) was added to the bark, and extracted at room temperature for 3 days and then filtered. To increase the extraction yield, two more methanol solutions were added, followed by extraction in the same manner and filtered. The filtered methanol extracts were combined and concentrated to 330 ml using a vacuum concentrator. 330 ml of methanol was added to the concentrated extract, and then 660 ml of heptane was mixed and the nonpolar material was removed for 2 hours in a 30 ° C mixed shake water bath. Two more times in the same manner using heptane to remove the nonpolar material. To further remove the nonpolar material, 300 ml of diethyl ether was added to the methanol extract, and the nonpolar material was removed for 2 hours in a 30 ° C mixed shake water bath. The heptane was used twice more in the same manner to further remove the nonpolar material. The methanol layer from which the non-polar material was removed was removed by methanol concentration, and the remaining aqueous layer was dried using a spray dryer to finally obtain 2.48 g of an antioxidant powder.

Experimental Example

1) Samples used for measuring antioxidant activity: The powdered pine bark-derived antioxidant samples obtained in Examples 1 to 8 and Comparative Examples 1 and 2 were dissolved in methanol for each concentration (eg, 50, 100, 300, 500, 700 μg / ml).

2) Antioxidant activity measurement method: The antioxidant activity of the sample was investigated by measuring DPPH (1,1-diphenyl-2-picrylhydrazine) radical scavenging activity and superoxide radical scavenging activity. At this time, Trolox, a water-soluble compound of BHA and vitamin E, which are representative antioxidants, was used as an indicator of activity test.

A) DPPH radical scavenging activity assay

Using a 96-well plate, 10 μl of each sample solution was added to each well, 90 μl of 1.5 x 10-4 M DPPH solution dissolved in ethanol was added and allowed to react for 10 minutes, followed by a microplate measuring instrument at 517 nm. Absorbance was measured. DPPH radical scavenging activity according to the concentration was calculated based on the following equation. The absorbance of the sample added with ethanol instead of the DPPH solution was blank, and methanol was used as a control.

DPPH radical scavenging activity (%) = (absorbance change of sample measured after 10 minutes) x 100 / absorbance of control

B) Determination of Superoxide Radical Scavenging Activity

Superoxide radical scavenging activity was measured using Riboflavin / EDTA / Nitroblue Tetrazolium (NBT) method. Using a 96-well plate, add 90 μl of 0.03 mM Riboflavin, 1 mM EDTA, 0.6 mM Methionine, and 0.03 mM NBT solution dissolved in 50 mM Potassium Phosphate Buffer (pH 7.8) per well and 10 μl of each sample solution. The activity of scavenging the superoxide generated by the reaction was measured. The superoxide-generating photochemical reaction was carried out using two fluorescent lamps installed in a light irradiation box made of aluminum foil, and the distance between the lamp and the reaction vessel was adjusted to 1000 Lux. It was measured using a microplate reader. The superoxide radicals generated by the photochemical reaction react with NBT to form a blue formazan, which increases the light absorption. When the superoxide radical is eliminated by the sample, the formation of the formazan decreases and the light absorption is reduced. Therefore, the superoxide scavenging activity of the sample was calculated according to the following equation using methanol as a control.

Radical scavenging activity (%) = (1-absorbance change of sample) x 100 / absorbance change of control

From the absorbance results of each sample obtained in the DPPH radical scavenging activity and the superoxide radical scavenging activity, the concentration of the sample required to inhibit radical formation by 50% is expressed as an ED50 (μg / ml) value, and the antioxidant activity of each sample is determined by measuring method (Table 1 and Table 2) were compared, and the total amount of antioxidant activity (extraction yield x relative antioxidant activity) in terms of total amount of antioxidant activity of each sample was compared (Table 3).

The lower the ED50 value of the sample, the higher the antioxidant activity of the sample. As shown in Table 1, the extraction yield of the antioxidant according to the present invention was 4.6 to 6.4%, and the maximum yield was 21 times higher than that of US Patent No. 4,698,360 (0.21%), and Korean Patent No. 10-0212974 (2.5%). The extraction yield was about 2.6 times higher. In addition, as shown in Table 2, in the case of the antioxidant activity against DPPH radicals, the antioxidant material produced by the present invention showed better results than the conventional methods, and showed about 1.1 to 1.7 times the antioxidant activity compared to Trolox. In the case of antioxidant activity against superoxide, the antioxidant prepared by the present invention showed similar antioxidant activity to the sample according to US Pat. No. 4,698,360, but showed about 1.7 times higher antioxidant activity than the sample according to Korean Patent No. 10-0212974. In particular, it showed a very strong superoxide scavenging antioxidant activity of about 1.7-2.2 times compared to BHA and 27-35 times higher than Trolox. Therefore, as shown in Table 3, the antioxidant material prepared by the present invention has significantly higher extraction yield and antioxidant activity than conventional methods, and when converted to the total amount concept multiplied by the extraction yield and antioxidant activity, DPPH compared to conventional control methods Antioxidant activity according to the present invention showed a total amount of antioxidant activity of at least 3.5 times to 45 times according to the antioxidant activity measurement method, and at least 2.6 times to 30 times according to the Superoxide antioxidant activity measurement method. The economic feasibility of the

 Comparison of Extraction Yield and DPPH Scavenging Activity of Antioxidants sample Extraction yield (%) DPPH clearing activity (ED50, μg / ml) Relative antioxidant activity (%), compared to Trolox BHA - 6.5 392 Trolox - 25.5 100 Example 1 5.8 21.0 121 Example 2 5.4 17.5 146 Example 3 5.0 23.5 109 Example 4 5.5 16.8 152 Example 5 5.0 24.2 105 Example 6 4.6 15.3 167 Example 7 6.4 22.5 113 Example 8 5.5 20.3 126 Comparative Example 1 0.21 29.0 88 Comparative Example 2 2.5 42.5 60

 Comparison of Antioxidant Superoxide Eliminating Antioxidant Activity sample Superoxide Scavenging Activity (ED50, μg / ml) Relative antioxidant activity (%), compared to BHA Relative antioxidant activity (%), compared to Trolox BHA 12.7 100 1,575 or more Trolox More than 200 Less than 6.4 100 Example 1 6.8 187 2,940 or more Example 2 6.2 205 More than 3,230 Example 3 7.3 174 2,740 or more Example 4 5.7 223 3,510 or more Example 5 7.2 176 2,780 or more Example 6 5.9 215 More than 3,390 Example 7 7.5 169 2,640 or more Example 8 6.9 184 2,880 or more Comparative Example 1 6.4 198 More than 3,390 Comparative Example 2 9.5 134 2,110 or more

Comparison of Antioxidant Activity by Manufacturing Method Applying Total Amount of Antioxidant Activity Sample or Sample Preparation Method Total amount of antioxidant activity by DPPH method Comparison of relative gross activity by DPPH method (fold) Total Antioxidant Activity by Superoxide Method Comparison of Relative Total Amount Activity by Superoxide Method Example 1 707 38.3 977 26.1 Example 2 789 42.7 1,024 26.6 Example 3 547 29.6 849 21.1 Example 4 840 45.5 996 29.6 Example 5 523 28.3 794 21.0 Example 6 772 41.8 872 23.9 Example 7 730 39.5 933 26.2 Example 8 695 37.7 1,095 24.4 Comparative Example 1 18 1.0 ** 42 1.0 ** Comparative Example 2 149 8.1 332 8.0

* Total amount of antioxidant activity = Extraction yield X Relative antioxidant activity (Apply data from Table 1 and Table 2)

** Calculates Comparative Example 1 as 1.0 in comparing relative total activity of antioxidant activity

As described above, the present invention extracts an antioxidant from pine bark using a fermentation alcohol solution, which is a relatively chemically stable physical solvent, and extracts such as rosin and the like using fermentation alcohol solution and / or natural animal and vegetable oils from the extract. Through the manufacturing process characterized by separating the non-polar material can produce an antioxidant having a high extraction yield and a high antioxidant activity and can provide an eco-friendly manufacturing process that is very easy to work.

The manufacturing process according to the present invention can increase the extraction yield of antioxidants at least 2.6 to 21 times as compared to the conventional control method, and also has an excellent antioxidant activity than the antioxidants prepared by the conventional methods in the antioxidant activity of the extract. Seemed. This is because the conventional methods are composed of a complex process of several steps, causing a lot of antioxidants lost during the process, and also due to applying the extraction separation process of non-polar materials having low efficiency. When converting the total amount of antioxidants in terms of the total amount multiplied by the extraction yield and the antioxidant activity of the extract, the method for preparing an antioxidant according to the present invention provides an antioxidant extract of at least 2.6 times and up to 45 times higher than conventional methods.

In addition, in the conventional method, various problems caused by using various synthetic organic solvents; In other words, fire hazard due to high volatility of solvent, worker safety, waste disposal cost, solvent reprocessing cost, restriction of food use due to organic solvent remaining in extract, but the present invention is harmless natural fermentation used as food The use of alcoholic solutions and natural animal and vegetable oils solves these problems, making the manufacturing process relatively simple and safe, producing less environmental pollutants, and making the antioxidant extracts ready to be used as food, superior activity and price. It can provide competitive natural antioxidants and their manufacturing process. In addition, the extract according to the present invention can be used as additives or raw materials for cosmetics, pharmaceuticals or feed.

In addition, by using domestic pine bark, which can be obtained as a by-product of domestic field and timber industry, it is possible to manufacture and supply highly active natural antioxidants at low cost, thus recycling domestic forest by-products, increasing the added value of pine trees, and In addition to increasing income, we also contribute to the development of the food industry, improvement of national health and quality of life through the development of various functional foods, processed foods, and medicines using the produced antioxidants. It is a useful invention that can greatly contribute to improving the balance of payments

Claims (9)

Extracting an antioxidant from pine bark using a ethanol solution which is a natural solvent; Separating the non-polar substances from the primary extract using a natural alcohol solution and / or natural animal and vegetable oil for the primary extract, pine bark derived natural antioxidants. According to claim 1, wherein the alcohol solution is characterized in that the ethanol concentration of 1.0 ~ 95% fermentation alcohol, pine bark derived natural antioxidants extraction method. According to claim 1, The natural animal and vegetable oil is soybean oil (soybean oil), corn oil (jade oil), rapeseed oil, rice bran oil, sesame oil, perilla oil, safflower oil, sunflower oil, cottonseed oil, peanut oil, olive oil, palm oil, palm oil, mixed cooking oil , Refined common oil, shortening, red pepper seed oil, tallow and lard selected from the group consisting of two or more oil, pine bark-derived natural antioxidants extraction method. The pine bark of claim 1 is pine, red pine (Pinus densiflora Sieb & Zucc.), Sea pine (Pinus thumbergii Parlatore), pine tree (Pinus densiflora Sieb. & Zucc. For. Erecta Uyeki), old tree L.), pine bark (Pinus pinea, Cupressus Sempervirens), causarina equisetifolia JR & G. Forst., Casuarina litorea L. Casuarinaceae, and Abies pindrow. A method for extracting natural antioxidants derived from pine bark, characterized in that it is applied alone or two or more. According to claim 3, The amount of natural animal and vegetable oil is used to extract non-polar substances by using a weight ratio of 0.5 to 20 times the extract concentrate or dried extract, pine bark-derived natural antioxidants extraction method . The method of extracting natural antioxidants derived from pine bark according to claim 1, wherein the antioxidants are separated by separating the non-polar substances from the primary extract using both the natural alcoholic solution and the natural animal and vegetable oil. delete delete delete