KR20200114406A - ANT-INFLAMMATORY COMPOSITION AND ITS MANUFACTURING METHOD USING Zanthoxylum schinifolium OIL - Google Patents

Abstract

Disclosed are an anti-inflammatory composition of Zanthoxylum schinifolium oil according to a pretreatment process, and a method for manufacturing the same. The method for manufacturing the anti-inflammatory composition of Zanthoxylum schinifolium oil according to the pretreatment process of the present invention comprises: a drying step of drying fruits of Zanthoxylum schinifolium; a separation step of separating the dried fruit into pericarp and seeds; a treatment step of treating seeds by using a method selected from using seeds as they are, roasting, and steaming the same; and an oil expression step of pulverizing seeds treated in the treatment step, followed by extracting oil by pressing.

Description

Anti-inflammatory composition of Japanese pepper oil according to pretreatment process and its manufacturing method {ANT-INFLAMMATORY COMPOSITION AND ITS MANUFACTURING METHOD USING Zanthoxylum schinifolium OIL}

The present invention relates to an anti-inflammatory composition and a method for producing the same, and more particularly, to an anti-inflammatory composition and a method for producing the same, using pepper oil prepared by a pretreatment process.

Sancho ( Zanthoxylum schinifolium ) is a deciduous shrub belonging to the Unhyang family and is a fragrant plant resource. It grows widely in the south of the central part of Korea, China, Japan, Taiwan, Manchuria, etc. It is a crop that grows well in sunny places all over the country except Hamgyeongbuk-do.

Sancho has many names such as japi, gpi, Namcho, Jincho, Bancho, and Chokcho. Sancho includes limonene, citronellal, phellandrene, sanshool, and flavonoids. ) Contains a lot of ingredients.

The fruit of the Japanese pepper tree is a light green fruit, round and blue, and the oil extracted from the fruit seeds has high essential oil content, so it has been used for gastrointestinal diseases and bronchial asthma since ancient times, and is useful for dry stomach, anti-inflammatory, diuretic, anthelmintic, gastric dyspepsia, and stomach enlargement. It is used as a treatment for anorexia, neuralgia, toothache, hypotension, refrigeration, bulimia, urolithiasis, antidiarrheal cold, and asthma.

The above-described technical configuration is a background technology for aiding understanding of the present invention, and does not mean a conventional technology widely known in the art to which the present invention belongs.

Korean Registered Patent Publication No. 10-1673202 (Wonkwang University Industry-Academic Cooperation Foundation) 2016. 11. 01.

Therefore, the technical problem to be achieved by the present invention is to provide an anti-inflammatory composition of sancho oil according to a pretreatment process that can utilize acidic oil as an anti-inflammatory composition, and a method of manufacturing the same.

According to an aspect of the present invention, a drying step of drying the fruit of the Japanese pepper tree; A separation step of separating the dried fruit into pericarp and seeds; And a treatment step of treating the seed as it is with a selected one of a stir-fry treatment and a steaming treatment. And a milking step of pulverizing and milking the seeds treated in the treatment step. A method of preparing an anti-inflammatory composition of peppercorn oil according to a pretreatment process may be provided.

In the drying step, the fruit of the sancho tree may be dried in a ratio of 11 to 12% based on the moisture weight of the sancho tree.

The stir-fry treatment may be treated by roasting the seeds at 80 to 100°C for 17 to 23 minutes.

The steaming treatment may be prepared by steaming the seeds for 27 to 32 minutes.

In addition, according to another aspect of the present invention, as an anti-inflammatory composition of sancho oil, an anti-inflammatory composition of sancho oil according to a pretreatment process comprising sancho oil prepared by any one of the above-described methods may be provided.

Embodiments of the present invention can be used as an anti-inflammatory composition after extracting the sancho oil from the sancho seed, which is a food material having various physiological activities, as it is used, roasting, and steaming.

1 is a diagram schematically showing a method of preparing an anti-inflammatory composition of sancho oil according to a pretreatment process according to an embodiment of the present invention.
FIG. 2 is a diagram schematically showing the acid value of sancho oil prepared by the method shown in FIG. 1.
FIG. 3 is a diagram schematically showing the high oxide value of the oxidative oil produced by the method shown in FIG. 1.
4 is a diagram schematically showing the cell viability of the sancho oil prepared by the method shown in FIG. 1.
FIG. 5 is a diagram schematically showing the production of nitrogen monoxide from the sancho oil prepared by the method shown in FIG. 1.

In order to fully understand the present invention, operational advantages of the present invention, and objects achieved by the implementation of the present invention, reference should be made to the accompanying drawings illustrating preferred embodiments of the present invention and the contents described in the accompanying drawings.

Hereinafter, the present invention will be described in detail by describing a preferred embodiment of the present invention with reference to the accompanying drawings. The same reference numerals in each drawing indicate the same member.

1 is a diagram schematically showing a method of preparing an anti-inflammatory composition of sancho oil according to a pretreatment process according to an embodiment of the present invention.

As shown in this figure, the manufacturing method of the anti-inflammatory composition of peppercorns oil according to the pretreatment process according to the present embodiment includes a drying step (S10) of drying the fruit of the peppercorns, and the dried fruit is applied to the pericarp and seeds. Separation step (S20) of separating into, a treatment step (S30) of treating the seed with a selected one of use as it is, roasting treatment and steaming treatment, and a milking step (S40) of pulverizing and milking the seeds treated in the treatment step (S30). ).

The drying step (S10) is a step of drying the fruit of the Japanese pepper tree before separating it into pericarp (the part surrounding the fruit seed) and seeds.

In the present embodiment, the drying step (S10) may be provided by drying the fruit of the sancho tree at a ratio of 11 to 12% of the moisture weight of the sancho tree.

The separation step (S20) is a step of separating the fruit of the Japanese pepper tree dried in the drying step (S10) into pericarp and seeds.

In this embodiment, the seeds separated in the separation step (S20) may be made of sancho oil.

The treatment step (S30) is to use the seeds separated in the separation step (S20) as they are, that is, use (no treatment) without going through a separate treatment step (S30), roast the separated seeds, or steam the separated seeds ( Steaming treatment) can be made in one of the methods selected.

In the treatment step (S30), when the separated seeds are used as they are to make sancho oil, there is an advantage in that the rancidity progress rate is low as described below compared to the roasting treatment or steaming treatment.

The stir-fry treatment during the treatment step (S30) may be treated by roasting the separated seeds at 80 to 100°C for 17 to 23 minutes. Roasting seeds separated by the above-described temperature and time has the advantage of increasing oxidation stability as described later.

During the treatment step S30, the steaming treatment may be prepared by steaming the separated seeds for 27 to 32 minutes. Steaming the separated seeds for the aforementioned time has the advantage of suppressing the generation of nitrogen monoxide after the no-treatment method.

In this embodiment, the sancho oil prepared by the above-described method is mixed with treatments such as dry stomach, anti-inflammatory, diuretic, anthelmintic, gastric hydration, anorexia, neuralgia, toothache, hypotension, refrigeration, bulimia, urolithiasis, antidiarrheal cold, asthma, etc. It can be prepared as an anti-inflammatory composition.

FIG. 2 is a diagram schematically showing the acid value of the acidulant oil prepared by the method shown in FIG. 1, and FIG. 3 is a view schematically showing the high oxide value of the acidulant oil produced by the method shown in FIG. 4 is a diagram schematically showing the cell viability of the sancho oil prepared by the method shown in FIG. 1, and FIG. 5 is a diagram schematically showing the production of nitrogen monoxide in the sancho oil prepared by the method illustrated in FIG. 1.

The above-described method, namely, acid value, peroxide value, fatty acid analysis, RAW264.7 cell culture, cytotoxicity measurement (MTS assay), and measurement of Nitric oxide (NO) production of untreated, roasted, and steamed pepper oil are as follows.

The acid value is analyzed by modifying the AOCS method (2007). Take 1 g of an untreated, stir-fried, or steamed acidic oil fat and oil sample into an Erlenmeyer flask, and add 30 mL of a mixed solution of ethanol: ether (1:1 v/v) to dissolve it. Add 100 uL of 1% phenolphthalein solution as an indicator, and titrate with 0.1 N KOH (in ethanol) solution until the end point (pale red) is reached. A blank test was conducted without a sample, and the acid value was calculated by substituting it into this equation.

Acid value (mg KOH/g oil) = 5.611 ×(a-b) ×f / S

a: Consumption amount of 0.1 N KOH/ethanol standard solution of this test (mL)

b: Consumption of 0.1 N KOH/ethanol standard solution in blank test (mL)

f: Potency of 0.1 N KOH/ethanol standard solution (factor)

S: sample weight (g)

Acid value is an essential item in determining the quality of oils and fats and foods containing oils and fats, and in particular, it is a standard indicating the degree of rancidity of oils and fats by measuring the amount of free fatty acids in which fatty acids are not bound to glycerides. If the storage condition of food fats and oils is poor, rancidity occurs in which fatty acids are released by hydrolysis or oxidation, so it can be seen that the quality of fats and oils decreases as free fatty acids increase.

As shown in FIG. 2, the acid value was the lowest at 1.65±0.29mg/g in untreated sancho oil, 2.84±0.24mg/g in steamed processed sancho oil, and 3.47±0.26 in roasted processed sancho oil. It appears somewhat higher in the order of mg/g.

Therefore, the acid value indicating the degree of rancidity of fats and oils is that untreated processed sancho oil has a lower rate of rancidity than steamed and roasted processed sancho oil.

The peroxide value is analyzed by modifying the AOCS method (2007). To a 250 mL Erlenmeyer flask, add 25 mL of a mixed solution of acetic acid: chloroform (3:2 v/v) to 2 g of the acidic oil maintenance sample and dissolve. After that, 1 mL of saturated KI solution is added, mixed, and left in the dark for 10 minutes. Add 75 mL of distilled water and 1 mL of 1% soluble starch solution, and titrate with 0.01 N sodium thiosulfate (Na ₂ S ₂ O ₃ ) solution until the end point (colorless). The blank test proceeds without processing the sample, and the peroxide value is calculated by substituting it into this equation.

Peroxide value (meq/kg) = (a-b) ×0.01 ×f ×1000 / S

a: Consumption amount of 0.01 N Na2S2O3 standard solution of this test (mL)

b: Consumption of 0.01 N Na2S2O3 standard solution in blank test (mL)

f: The titer of 0.01 N Na2S2O3 standard solution (factor)

S: sample weight (g)

Peroxide value refers to the number of mg equivalents of peroxide contained in 1 kg of oil and fat, and is an index indicating the degree of rancidity in the initial stage of oil and fat. However, since it indicates the initial rancidity of fats and oils, the higher the peroxide value, the less fresh, and after the content of hydroperoxide generated from the rancidity of fats and oils reaches the highest level, peroxides are carbonyl compounds such as aldehydes and ketones. Since it decomposes into, the peroxide value tends to be low for oils that have passed a long time since rancidity occurred.

As shown in Fig. 3, the peroxide value of the peppercorn oil according to the pretreatment processing process is the lowest at 144.27±3.24mg/g in the untreated peppercorn oil, and 156.44±1.57mg/g in the steamed peppercorn pepper oil, as shown in FIG. It appears somewhat higher in the order of 188.38±1.92mg/g in processed pepper oil. Therefore, the peroxide value, which indicates initial rancidity, is less likely to be initially oxidized than untreated and roasted sancho oil.

Fatty acid composition analysis is analyzed by modifying the AOCS method (1989). In a glass test tube, add about 100 mg of acido oil and 500 uL of fatty acid internal standard heptadecanoic acid (C17:0; 1 mg/mL hexane) and then 0.5 N NaOH/methanol solution 2 mL Add to melt. After that, heat it for 10 minutes using a heating block heated to 100°, cool it on ice, and add 4 mL of 14% BF3-methanol to derivatize it in a 100° heating block for 40 minutes. After cooling the derivatized sample on ice, 2 mL of hexane and 5 mL of saturated NaCl were added, rotated for 2 minutes, and a portion of the supernatant was filtered through a 0.45 μm hydrophobic membrane filter to analyze fatty acids. Use as a sample.

Fatty acid was analyzed using gas chromatography (GC; Agilent 7693A, Agilent Co., Palo Alto, CA, USA), and the analysis conditions are shown in Table 1 below.

Instrument Condition Model Gas chromatography 7693A (Agilent Co., USA) Column DB-waxetr column
30 mⅹ0.25 mm, id,0.25-㎛ film thickness (Agilent Co., USA) Oven Temperature 100°-2min -3°-240°-30℃/min -260℃ -5min Injector Splitless mode, 240° Detector Flame ionization detector, 260° Carrier gas N2 20 mL/min Injection volume 1 μL

The average content of fatty acids in the pepper oil according to the pretreatment process is as follows.

The average content of major fatty acids in untreated sancho oil is palmitic acid (C _16:0 ) 3.09%, margaric acid (C _17:0 ) 11.65%, oleic acid (C _18:1 ) 11.20%, linoleic acid (C _{18: 2} ) 44.27%, gamma-linolenic acid (C _18:3 ) 28.58%, arachidic acid (C _20:0 ) 1.21%, and the average content of major fatty acids in roasted pepper oil is palmitic acid (C _16:0 ) 3.57%, margaric acid (C _17:0 ) 14.51%, oleic acid (C _{18: 1} ) 12.76%, linoleic acid (C _18:2 ) 43.41%, gamma-linolenic acid (C _18:3 ) 24.56%, arachidicdic acid(C _20:0 ) 1.20%, the average content of major fatty acids in steamed pepper oil is palmitic acid (C _16:0 ) 3.06%, margaric acid (C _17:0 ) 11.89%, oleic acid (C _18:1) ) 10.72%, linoleic acid (C _18:2 ) 43.79%, gamma-linolenic acid (C _18:3 ) 29.36%, arachidic acid (C _20:0 ) 1.19%.

In terms of fatty acid composition, unsaturated fatty acids such as oleic acid, linoleic acid, and gamma-linolenic acid appear in the order of 84.05%, 83.86%, and 80.73%, respectively, as untreated, steamed, and roasted sancho oil, respectively, and saturated fatty acids are roasted. The composition is 19.27%, 16.14%, and 15.95%, respectively, with treatment processing, steaming processing processing, and no processing processing. This is because the roasting process has high oxidation stability, but the untreated processed pepper oil contains a lot of unsaturated fatty acids, which are essential fatty acids, so it is high in terms of nutrition and functionality.

RAW264.7 cells, a macrophage cell line of the mouse used in this experiment, were DMEM (Dulbecco'modified eagle medium) medium with 10% FBS (fetal bovine serum) and 1% P/S (penicilin-streptomycin) added for cell culture. Use. When the cultured RAW264.7 cells are 80% confluent, the cells are detached using a Cell Scraper, centrifuged at 4,000 rpm for 4 minutes, and then subcultured every 2 days.

The cytotoxicity of RAW264.7 to pepper oil according to the pretreatment process was measured by slightly modifying the method of 5-(3-carboxyme thoxyphenyl)-2H-tetraza lium inner salt (MTS). Dispense RAW264.7 cells into a 96-well plate at 2*10 ⁴ cells/well and incubate in an incubator at 37°C and 5% CO ₂ for 24 hours. Thereafter, the samples were treated at different concentrations (10, 50, 100, 200, 500 μg/mL) to RAW264.7 cells and cultured for 18 hours. Thereafter, 20µl of MTS solution per well is added, reacted for 2 to 3 hours in an incubator, and absorbance is measured at 490 nm using a microplate reader (SpectraMax M5, Molecular Devices, CA, USA).

The results of measuring the cell viability of RAW 264.7 macrophages by MTS assay by sancho oil according to the pretreatment processing process are shown in FIG. 4 and are as follows. The 5-(3-caroboxymeth-oxyph enyl)-2H-tetra-zolium inner salt (MTS) assay is a method of measuring the conversion of MTS to formazan by mitochondrial dehydrogenases. RAW 264.7 macrophages were treated with fermented mixtures for each concentration of medicinal plants, cultured for 24 hours, and then treated with MTS. As a result, when the survival rate of the control group not treated with sancho oil according to the pretreatment process was 100%, 500 ug It was confirmed that the cell viability was more than about 80% in the acidophilus oil, excluding the roasted processed acidophilus oil of /nL, and the acidophilic oil sample of 200 ug/mL or less did not show cytotoxicity to RAW 264.7 cells.

Therefore, in this experiment, it was determined that if the experiment was conducted at a sample treatment concentration of 200 μ or less of sancho oil, it would not have a significant effect on the experimental result, and the sample concentration was conducted at a sample concentration of 200 μ or less.

The concentration of NO is 2*10 ⁴ cells/well in a 96-well plate and incubated in an incubator under conditions of 37°C and 5% CO ₂ for 24 hours, and then sampled by concentration (10, 50, 100, 200). Μg/mL) 2 hours pre-treatment, then LPS (100 ng/mL) treatment and incubation for 18 hours. After adding the same amount of Griess reagent as the culture medium, after 5 minutes, the absorbance was measured at 540 nm using a microplate reader (SpectraMax M5, Molecular Devices, CA, USA). The concentration of NO is calculated based on the standard curve for each concentration of NaNO ₂ .

Nitric oxide (NO), a reactive nitrogen species (RNS) that appears in inflammatory reactions, is a free radical in an unstable gaseous state. As a transport material for the cardiovascular, nervous and immune systems, it maintains intracellular homeostasis, anticancer action, and transports neurotransmitters. And a substance having various functions such as cytotoxicity. On the other hand, when excessively generated, it acts as a toxic radical, activating the inflammatory reaction that causes damage to cells or tissues, as well as promoting the biosynthesis of inflammatory mediators to intensify inflammation.

The results of treating the cell with 100 ng/mL of LPS for 18 hours after two hours of pretreatment of the sancho oil according to the pretreatment processing process are shown in FIG. 5. In the group not treated with LPS, the concentration of NO was 3.28±1.01 μM, and in the group treated with LPS, the concentration of NO was 23.96±1.2 μM, which was higher than that of the sample treatment group. Sancho oil according to the pretreatment processing process showed high NO production inhibitory ability depending on the concentration, and untreated sancho oil was 8.17±2.04~15.63±0.79μM, showing the highest NO inhibition generation ability. The steaming-processed sancho oil shows 11.46±1.33~17.67±0.29μM, the stir-processed sancho oil 12.41±0.92~18.16±0.36μM in the order of NO production inhibition.

Therefore, sancho oil, which is milked by a non-treatment process, shows excellent anti-inflammatory effects, and is highly likely to be used as a material application value of sancho in anti-inflammatory related fields.

As described above, the present invention is not limited to the described embodiments, and it is obvious to those of ordinary skill in the art that various modifications and variations can be made without departing from the spirit and scope of the present invention. Therefore, it should be said that such modifications or variations belong to the claims of the present invention.

S10: drying step
S20: separation step
S30: processing step
S40: milking stage

Claims (5)

A drying step of drying the fruits of the Japanese pepper tree;
A separation step of separating the dried fruit into pericarp and seeds; And
A treatment step of treating the seeds as they are, using a stir-fry treatment, or a steaming treatment; And
A method for producing an anti-inflammatory composition of peppercorn oil according to a pretreatment process comprising a milking step of pulverizing and milking the seeds treated in the treatment step. The method according to claim 1,
The drying step is a method for producing an anti-inflammatory composition of sancho oil according to a pretreatment process, characterized in that drying the fruit of the sancho tree at a ratio of 11 to 12% of the moisture weight of the sancho tree. The method according to claim 1,
The stir-fry treatment is a method for producing an anti-inflammatory composition of sancho oil according to a pretreatment process, characterized in that the seeds are roasted at 80 to 100°C for 17 to 23 minutes. The method according to claim 1,
The steaming treatment is a method for producing an anti-inflammatory composition of sancho oil according to a pretreatment process, characterized in that the seeds are steamed for 27 to 32 minutes. As an anti-inflammatory composition of pepper oil,
An anti-inflammatory composition of peppercorn oil according to a pretreatment process comprising peppercorn oil prepared by the method of any one of claims 1 to 4.

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