Extract of Athyrium niponicum with antibacterial and antioxidant activity

专利摘要:
PURPOSE: An extract of Athyrium niponicum having antimicrobial and antioxidant activities is provided, thereby obtaining natural antimicrobial agent and antioxidant having improved activities. CONSTITUTION: The extract of Athyrium niponicum having antimicrobial and antioxidant activities is produced by the steps of: adding 6 liter of ethanol into 600 g of dried Athyrium niponicum and suspending them for 48 hours; filtering the suspension with a filter to obtain an ethanol fraction; adding 6 liter of ethanol into the remaining solution to obtain an additional ethanol fraction; concentrating the ethanol fraction under reduced pressure and freeze-drying the concentrate; dissolving the freeze-dried ethanol extract into 500 milliliter of a mixed solution of water and ethanol in the volume ratio of 9:1; adding 500 milliliter of chloroform into the ethanol extract-containing solution to obtain a chloroform extract; sequentially adding ethylacetate and butanol into the chloroform extract to obtain fractions of ethylacetate, butanol and water; and filtering and concentrating them under reduced pressure and freeze-drying the concentrates.
公开号:KR20020095146A
申请号:KR1020020074150
申请日:2002-11-20
公开日:2002-12-20
发明作者:박인호；송종호；김민주
申请人:박인호；
IPC主号:

专利说明:

Extract of Athyrium niponicum with antibacterial and antioxidant activity
[4] The present invention looks at the antimicrobial and antioxidant activity using the extract of fern. More specifically, the present invention is to obtain the fern extract sequentially with ethanol, chloroform, ethyl acetate, butanol and water using the polarity of the solvent and then to measure the antimicrobial function and antioxidant capacity to provide a material as an antibacterial and antioxidant agent .
[5] Most of the existing antimicrobials used in the past have been manufactured through chemical synthesis, ranging from pesticides to antibiotics used as medicines. However, the emergence of resistant bacteria to such chemical synthetic antimicrobial agent is a frequent occurrence, and most of these chemical synthetic antimicrobial agent itself has a high toxicity because it is harmful to copper and plants, the use of a chemical synthetic antimicrobial agent tends to be avoided.
[6] In addition, artificial synthetic antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate (PG), and tert-butyl hydroquinone (TBHQ), which are widely used to prevent long-term preservation and rancidity of foods, On the other hand, thermal stability is poor, and harmful substances are generated to raise safety issues. Therefore, the search for new safe natural antioxidants, especially antioxidants contained in foods, is required. Tocopherol is one of the most known natural antioxidants, and it is known to be very practical because it has the highest antioxidant capacity, and phenolic substances, ascorbic acid, carotenoids, sulfur compounds, and nitrogen compounds have been reported to have antioxidant properties.
[7] Food rancidity and oxidation in the body not only lowers food quality and nutritional value, but also causes various diseases and aging. Damage caused by harmful microorganisms is not only in the food industry, but also in medicine, agriculture, storage, distribution, etc. Since it is a serious problem facing a wide range of areas, there is a demand for the development of functional materials having natural antioxidant and antimicrobial activity that can ensure more stability.
[8] The present invention is to search for antibacterial and antioxidant functions using the extract of the fern to provide a functional material material as a natural antibacterial and antioxidant.
[1] 1 is a table evaluating the antimicrobial activity using the solvent extract of the dog fern.
[2] 2 is a table measuring the minimum growth inhibitory concentration of chloroform and ethyl acetate extract.
[3] 3 is a table measuring the antioxidant activity of the fern solvent extract.
[9] The present invention obtains the sequential extract using the fern native to Sanya of our country, search for the antimicrobial activity against various microorganisms and search the antioxidant function to evaluate the value as a food preservative to find the potential as a natural functional material to provide. Next, in order to more easily understand the present invention, preferred embodiments of the present invention are presented. The embodiments presented herein are merely presented to more easily understand the present invention, but the present invention is not limited thereto.
[10] Example 1. Preparation of sequential extract using dog fern
[11] Experimental material used in the present invention (Athyrium niponicum) using the outpost (basement) before the leaves grow, was identified using a plant illustration. 6 liters of ethanol was added to 600 g of dried dog fern, and then suspended at room temperature for 48 hours. The resultant was filtered through a filter paper to obtain an ethanol fraction, and the residue was re-extracted by adding 6 liters of ethanol to the residue. And lyophilized using a lyophilizer (F / D). The dried ethanol extract was redissolved in 500 ml of water: ethanol (9: 1), and 500 ml of chloroform was added thereto to obtain a chloroform extract. Ethyl acetate, butanol and water extracts were added sequentially to the aqueous fraction to obtain ethyl acetate, butanol and water extracts. Each sequential extract was filtered through a filter paper, concentrated under reduced pressure with a rotary vacuum condenser, and freeze-dried using F / D. Used as a material.
[12] Example 2. Determination of antimicrobial activity using ethanol and each sequential extract
[13] The antimicrobial activity of the five extracts obtained in Example 1 was determined by the method of disc diffusion of Bauer by selecting 20 pathogenic microorganisms (Am. J. Clin. Pathol., 45, 495, 1966). Each extract was re-dissolved in ethanol and dipped into sterilized discs (8mm, thin, Toyo Co., Japan) to concentrations of 0.25 and 0.5 mg / disc, and dried completely. 100 microliters of each strain cultured one day prior to antimicrobial activity assay were evenly applied to the plate medium, and the disc was completely adhered to the plate medium to which each test strain was applied, and in a 4 ° C. refrigerator for diffusion of each extract. After 1 hour of incubation, the presence and diameter of the zona pellucida around the disc were measured in mm units after 18 hours incubation at 37 ° C. The ethanol, chloroform and ethyl acetate extracts of dog fern showed antimicrobial activity among 16 assay bacteria among 20 assay bacteria at a dose of 0.25mg per disc, and antimicrobial activity was similar in size. The result of Example 1 is considered to be higher than the antimicrobial activity of other plant-derived natural substances.
[14] Example 3 Determination of Minimum Growth Inhibition Concentration of Chloroform and Ethyl Acetate Extract
[15] As a result of performing Example 2 using the five extracts extracted in Example 1, chloroform and ethyl acetate extracts having strong antibacterial activity were selected, and the method of Mann and Markham was modified to measure the minimum growth inhibitory concentration (J. Appl). Microbiol., 84, 538, 1998), inoculated with 25 μl of each assay suspended in a test tube containing 9.9 ml of medium and 100 μl of the extract at 660 nm for absorbance of 0.1 ± 0.02. Absorbance was measured at 660 nm after incubation in a shaker at 37 ° C., and the minimum concentration at which the absorbance was less than 0.1 and no growth of microorganisms was determined as the minimum growth inhibition concentration of microorganisms and is shown in FIG. 2. The minimum growth inhibitory concentration of chloroform and ethyl acetate extract was found to be 5-40 ppm. These results show antimicrobial activity comparable to that of grapefruit seed extract, which is widely used as a natural antimicrobial agent.
[16] Example 4. Determination of Antioxidant Activity of Ethanol and Each Sequential Extract
[17] Antioxidant activity was measured using 5 extracts extracted in Example 1 using 1,1-diphenyl-2-picrylhydrazyl (DPPH) reagent (Nature, 26, 1199, 1958). The DPPH solution was prepared by dissolving DPPH 16 mg in 100 ml ethanol and then mixing 100 ml of distilled water and filtering the resultant in a filter paper. After 30 minutes of mixing a 100 μl sample solution of a certain concentration into 4.9 ml of this solution, the decrease in absorbance was measured at 528 nm. At this time, the absorbance of the control without the sample was set to 100 and the value was reduced to 1/2, that is, the amount of the sample required to have RC50 (reduce concentration 50%) was measured. Shown in The ethanol extract showed 20 ~ 40ppm of RC50 value, 20ppm of chloroform extract, and 10 ~ 20ppm of ethyl acetate extract and butanol extract. The lower the value of RC50, the higher the antioxidant activity.
[18] In view of the antibacterial activity and antioxidant ability of the fern extract and sequential extract developed in the present invention, it will be possible to secure a substance that can enter the market as a new antibacterial and antioxidant. As a natural functional material, it is highly applicable to antibacterial purpose in crop cultivation field and antimicrobial preservative in food field, and it is also considered that it is possible to apply natural antimicrobial property to past cosmetics and new material in medicine field.

权利要求:
Claims (3)
[1" claim-type="Currently amended] Natural Composition Using Athyrium niponicum Extract and Sequential Extract
[2" claim-type="Currently amended] The composition according to claim 1, which has a property of antimicrobial activity.
[3" claim-type="Currently amended] The composition of claim 1 having properties of antioxidant function

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同族专利:

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公开号 | 公开日

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KR100483338B1|2005-04-18|

引用文献:

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公开号 | 申请日 | 公开日 | 申请人 | 专利标题

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法律状态:
2002-11-20|Application filed by 박인호

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2002-11-20|Priority to KR10-2002-0074150A

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2002-12-20|Publication of KR20020095146A

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2005-04-18|Application granted

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2005-04-18|Publication of KR100483338B1

优先权:

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申请号 | 申请日 | 专利标题

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KR10-2002-0074150A|KR100483338B1|2002-11-20|2002-11-20|Extract of Athyrium niponicum with antibacterial and antioxidant activity|