KR20160115311A - Antibacterial and antifungal composition comprising plant extracts - Google Patents
Antibacterial and antifungal composition comprising plant extracts Download PDFInfo
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- KR20160115311A KR20160115311A KR1020150042684A KR20150042684A KR20160115311A KR 20160115311 A KR20160115311 A KR 20160115311A KR 1020150042684 A KR1020150042684 A KR 1020150042684A KR 20150042684 A KR20150042684 A KR 20150042684A KR 20160115311 A KR20160115311 A KR 20160115311A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3472—Compounds of undetermined constitution obtained from animals or plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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Abstract
The present invention relates to a composition for antibacterial and antifungal use using a plant extract, and the composition for antibacterial and antifungal use according to the present invention is excellent in antimicrobial and antifungal effect against fungi including gram-positive bacteria and gram- And can be used more safely as a preservative in the pharmaceutical and food fields.
Description
The present invention relates to an antibacterial and antifungal composition comprising a plant extract.
Despite improvements in living environment and hygiene, food poisoning has been increasing in recent years due to global warming, rising indoor temperatures, mandatory school meals, and increasing eating out. As a result of epidemiological studies on food and drinking water, patient and food industry workers, more than 90% of food poisoning cases have been identified as bacterial and viral, and it is urgent to prepare for harmful factors of microorganisms rather than chemical hazards.
In order to prevent food poisoning caused by microorganisms and safely store food for a long period of time, there is a method of using physical sterilization methods such as heating, ultrahigh pressure, radiation, PEF and optical pulse, and chemical antimicrobial substances such as alcohol, chlorine, hydrogen peroxide, Due to problems such as convenience and cost, synthetic preservatives such as chemical antimicrobial substances, especially nitrite, sorbic acid, sodium metabisulfite, and chlorinated compounds have been steadily used. However, the synthetic preservative has not only been continuously posed safety problems such as accumulation in the human body, but has also been avoiding its use in the food industry due to the tendency of consumers to pursue well-being. Therefore, development of a natural antimicrobial substance with safety is required.
Natural antimicrobial substances that have been traditionally used include nisin derived from proteins or enzymes present in animals and plants, chitosan extracted from crustaceans, various organic acids derived from natural products, plant essential oils and microorganisms, as well as food materials such as salt and vinegar. e-polylysine and natamycin have been reported (Korea Patent No. 0123131 and Korean Patent No. 0025933), but natural antimicrobial substances have a characteristic taste, odor, Due to poor characteristics, problems such as low antibacterial activity and narrow antimicrobial spectrum, only a few commercialized cases are still present.
Accordingly, the inventors of the present invention have conducted extensive research to solve the problems of the prior art described above. As a result, the present inventors have completed the present invention by confirming that the extracts of domestic native plants have excellent antibacterial and antifungal activity.
Accordingly, an object of the present invention is to provide an antibacterial and antifungal composition comprising an extract of domestic native plants.
Another object of the present invention is to provide a natural preservative comprising the composition for antibacterial and antifungal.
In order to accomplish the above object, the present invention provides a method for producing a plant of the genus Siegesbeckia glabrescens , Largerstroemia indica , Spiraea trichocarpa , Rhus trichocarpa ), Rhus verniciflua ( Rhus succedanea ), turbid ( Hosta longipes ), large crane flowers ( Majanthemum dilatatum ) and cassia ( Cercidiphyllum a mixture of plant extracts selected from the group consisting of japonicum .
To achieve these and other objects, the present invention provides a natural preservative comprising the composition for antibacterial and antifungal use.
The antimicrobial and antifungal composition according to the present invention has excellent antimicrobial and antifungal effects against bacteria including bacterium including Gram-positive bacteria and Gram-negative bacteria, fungi including yeast and fungi, and can be safely used as a preservative in pharmaceuticals, cosmetics, .
Hereinafter, the present invention will be described in detail.
The present invention relates to a method for the treatment of diseases such as Siegesbeckia glabrescens , Largerstroemia indica , Spiraea trichocarpa , Rhus trichocarpa , Rhus succedanea , Hosta longipes , Majanthemum dilatatum ) And a plant extract selected from the group consisting of Cercidiphyllum japonicum .
The antidepressant used in the antimicrobial and antifungal composition according to the present invention is an annual herbaceous plant belonging to the Asteraceae family, and exhibits a blood pressure lowering effect due to its vasodilating action, and is useful as a medicament effective against limb paralysis, muscle skeletal pain, weakness in the back, knee, . Bongong is a deciduous broad-leaved arboreous tree belonging to Bacillus subtilis. It is mainly used for ornamental or gardening because its flowering period is long and its flowers are beautiful. The mane is a deciduous shrub belonging to the Rosaceae. It grows frequently in the mountains, and the roots have the effect of heat and convergence. It is a deciduous broad-leaved tree belonging to Rhus verniciflua. It causes serious inflammation when it touches the skin, and it is used as a medicinal material effective for gastrointestinal diseases, kidney stones, liver diseases and physiological impurities. Sumatranopsis is a deciduous broad-leaved arboreous tree, similar to lacquer trees, but the whole leaf is rather small, has no hairs, and grows mainly in the lower parts of warmer regions. Vivacea is a perennial herbaceous plant belonging to the lily family, which blooms in summer, and young leaves are eaten as herbs and cultivated for ornamental purposes. The large Grus bloom is a perennial herbaceous plant belonging to the lily tree. All plants except root are used as a medicinal substance called mulogecho. It has a hemostatic effect on urine bleeding, hematemesis and menstrual hyperplasia. Cassia is a deciduous broad-leaved arboreous tree belonging to the Cassia cunea, and is mainly used as a raw material for topical medicines, confectionery, cooking and spices.
The plant extract used as the active ingredient of the antibacterial and antifungal composition according to the present invention may be a plant leaf, a stem, a core material, a bark, a flower or a combination thereof. Preferably, the plant extract is selected from the group consisting of the above ground parts (stem, leaf, flower, and seed), bark of bongo tree, ground part of non-vertebrate, Tree bark and core material, flowers of large crane, or any mixture of these extracts.
The plant extracts may be used alone, but they may exhibit inhibitory activity against a broader range of microorganisms when used in combination.
In one embodiment according to the present invention, the plant extract may be a combination of a true-to-ground surface part, a rabbit ground part, and a folium lacquer tree stalk. In another embodiment according to the present invention, the plant extract may be a combination of balsam bark, folium lacquer tree stems and large crane flower flowers. In another embodiment according to the present invention, the plant extract may be a combination of a mane sprout leaf, a rabbit top part, a folium lacquer tree stalk and a large crane flower flower. In another embodiment according to the present invention, the plant extract may be a combination of a ginseng root stalk, a rabbit top part, a faltering stamina leaf stalk, and a large crane flower. In another embodiment according to the present invention, the plant extract may be a combination of phloem var. Lacquer leaf, rabbit top part, phloem lacquer trunk and large crane flower. In another embodiment according to the present invention, the plant extract may be a combination of cassia bark, rabbit top, phloem lacquer stem and large crane flower. In another embodiment according to the present invention, the plant extract may be a combination of a cedar wood core material, a rabbit ground part, a folium lacquer tree stalk and a large crane flower flower.
Each of the plant extracts described above can be prepared by a conventional extraction method using an extraction solvent. Examples of the extraction solvent include, but are not limited to, water, C 1 -C 4 alcohol, ethyl acetate or a mixed solvent thereof.
The plant extract may be extracted at a temperature of 90 to 100 DEG C for 2 to 6 hours when water is used as an extraction solvent, but is not limited thereto.
In addition, the plant extract may be extracted for 1 to 3 hours at a temperature of 20 to 40 ° C using a C 1 to C 4 alcohol, ethyl acetate or a mixed solvent thereof, but is not limited thereto.
In one embodiment of the present invention, the plant extract may be prepared through two extraction steps. That is, the extract may be a mixture of the first extract extracted first using the first extraction solvent and the second extract extracted using the second extraction solvent.
In one embodiment of the invention, the extraction process may be accompanied by sonication to enhance extraction efficiency. That is, ultrasonic treatment can be applied while extracting each plant with an extraction solvent. The ultrasonic treatment conditions are set at 130 W and 20 kHz at intervals of 20 seconds, but the present invention is not limited thereto. Conventional conditions used for plant extraction may be applied.
The composition for antibacterial and antifungal use according to the present invention may contain a mixture of the above extract in an amount of 1 to 10% by weight based on the total amount of the composition. When the mixture of the extracts is used in an amount of 3 wt% to 7 wt% based on the total amount of the composition, it may exhibit an excellent antimicrobial activity that the present invention desires to achieve.
The antimicrobial and antifungal composition according to the present invention may be adjusted to have a pH ranging from 2.5 to 4.0 in order to maximize the antimicrobial effect. The pH range may be adjusted using an organic acid selected from the group consisting of organic acids such as citric acid, malic acid, lactic acid, tartaric acid, and combinations thereof
The antimicrobial and antifungal compositions according to the present invention exhibit inhibitory activity against bacteria (gram-positive and gram-negative) and fungi including yeast and fungi.
Examples of bacteria that can be inhibited by the composition of the present invention include bacteria such as Bacillus cereus , Enterobacter aerogenes , Listeria monocytogenes , Staphylococcus epidermidis ), Streptococcus mutans mutans , Escherichia coli , Proteus vulgaris , Salmonella enterica subsp. Enterica , Salmonella enterica, enteritidis , Vibrio parahaemolyticus and Vibrio vulnificus .
Examples of fungi that can be inhibited by the composition of the present invention include yeasts and fungi. Examples of yeasts include Candida albicans , Debaryomyces sp. hansenii there is . hansenii , and Pichia anomala . Examples of fungi include Aspergillus flavus , Mucor, circinelloides , Penicillium digitatum ) and Rhizopus oryzae .
The antimicrobial and antifungal composition according to the present invention has excellent antimicrobial and antifungal effects against bacteria, yeasts and fungi including Gram-positive bacteria and Gram-negative bacteria, and thus can be safely used as a preservative in pharmaceutical and food fields.
In addition, the present invention provides a natural preservative comprising the above antibacterial and antifungal composition.
The natural preservative according to the present invention can prevent deterioration and decay of food and increase preservation ability by killing harmful microorganisms and inhibiting proliferation.
[Example]
Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.
Example 1: Preparation of plant extracts
<1-1> Manufacture of extracts from bamboo trees, bamboo trees, mane japonks, lacquer trees, foxtail lacquer, Viviculae, big crane flowers and Cassia
Plant samples purchased from the market in the form of buildings were crushed into powder by a dry mill. To 10 g of the powder was added 200 ml of water and then heated at 100 ° C for 4 hours to prepare an extract of each plant. Thereafter, the extract was filtered and lyophilized using a freeze dryer to prepare a powdery plant extract.
<1-2> Manufacture of extracts from bamboo trees, bamboo trees, mane japonks, lacquer trees, foxtail lacquer, vivaceous, big crane flowers and cassiae
Plant samples purchased from the market in the form of buildings were crushed into powder by a dry mill. Ethanol (200 ml) was added to 10 g of the powder, and a primary extract of each plant was prepared using an ultrasonic shredder (Vibra Cell, Sonics and Materials Inc. USA) at a frequency of 130 W and 20 kHz every 20 seconds. Thereafter, the primary extract was filtered, and 200 ml of ethanol was further added to the filtrate residue to prepare a secondary extract of each plant in the same manner. Then, the above-prepared primary and secondary extracts were mixed, filtered and lyophilized using a freeze dryer to prepare a powdery plant extract.
<1-3> Manufacture of extracts from Guanxia province, Bongo tree, Mannoptera, Lacquer tree, Fagaceae lacquer, Viviculae, Big crane and Cassia
Plant extracts were prepared in the same manner as in Example 1-2 except that methanol was used instead of ethanol as the extraction solvent.
<1-4> Manufacture of extracts from natural sources, bongo tree, maneoptera, lacquer tree, fenugreek var.
The plant extract was prepared in the same manner as in Example 1-2 except that ethyl acetate was used instead of ethanol as the extraction solvent.
Test Example 1: Measurement of antimicrobial activity of plant extracts
The antimicrobial activity of the plant extract prepared in Example 1 was analyzed using an agar plate culture diffusion method. To the sterilized agar medium cooled to 40 ° C, 1 μl / ml of each microbial culture listed in the following Table 1 was added and mixed, and 20 ml of each was added to the petri dish to prepare an agar plate culture medium. Rounded grooves having a diameter of 9 mm were formed on the prepared agar plate medium using a sterilized stainless steel pipe. Each of the plant extracts diluted with 1% (W / V) ≪ / RTI > for 24 hours. After the incubation, the diameter of the clear zone of the microorganism growth which did not proliferate due to the diffusion of the antimicrobial substance around the grooves of each agar plate culture medium was measured using Scan 1200 (Inter Science, France) Respectively. The results are shown in Tables 1 and 2 below.
Above ground
fell
Above ground
Tree leaf
stem
Ram
amount
castle
three
Germ
Ram
Well
castle
three
Germ
mother
phut
this
stem
leaf
fell
Core material
Flower
Ram
amount
castle
three
Germ
Ram
Well
castle
three
Germ
mother
phut
this
As shown in Tables 1 and 2, the plant extract according to the present invention showed excellent antibacterial and antifungal activity against a wide range of microorganisms. Especially, the extracts of P. japonicus, C. japonica, and C. japonica showed excellent antimicrobial activity against E. coli among Gram-negative bacteria. And has excellent antifungal activity.
Example 2: Preparation of a mixture of plant extracts
The plant extracts prepared in Example 1 were mixed in various combinations and ratios as shown in Table 3 below to prepare mixtures.
Test Example 2: Antimicrobial and antifungal activity measurement of a mixture of plant extracts
The mixture of the plant extracts prepared in Example 2 was diluted to a concentration of 1% (W / V), and antibacterial activity and antifungal activity were measured in the same manner as in Test Example 1. [ The results are shown in Table 4.
Ram
amount
castle
three
Germ
Ram
Well
castle
three
Germ
mother
phut
this
As shown in Table 4, it was confirmed that the mixture of the plant extracts prepared in Example 2 had excellent antibacterial and antifungal activity against all 18 species including Gram-positive bacteria, Gram-negative bacteria and fungi.
Test Example 3: Measurement of antimicrobial activity of a mixture according to plant extract concentration
(W / V), 2.5% (W / V), and 5.0% (W / V) of the plant extract prepared in Example 2 were tested for antimicrobial activity and antifungal activity, , Antibacterial activity and antifungal activity were measured for the three microorganisms in the same manner as in Test Example 1. [ The results are shown in Table 5.
As shown in Table 5, it was confirmed that the antimicrobial activity was increased in proportion to the concentration of the plant extract mixture.
Based on the above results, the concentration of the plant extract mixture was fixed at 5%.
Test Example 4: Measurement of antimicrobial activity of mixed composition having different pH
The pH of the mixture of plant extracts prepared in Example 2 was adjusted to pH 3.5, pH 5.0 and pH 6.5 using citric acid, respectively, in order to examine the antibacterial and antifungal activity according to pH. Thereafter, the antimicrobial activity was measured in the same manner as in Test Example 1. The results are shown in Table 6.
As shown in Table 6, the antimicrobial activity of each of the mixed compositions was significantly increased as the pH was lowered. However, the antimicrobial activity against fungi was not significantly different from that of bacteria. Therefore, it was confirmed that the inhibition of growth by pH was more affected by bacteria than fungi.
Test Example 5: Measurement of antimicrobial activity of mixed composition according to kinds of organic acid and concentration
(W / V) and 0.5% (W / V) of citric acid, malic acid and lactic acid were added to the mixture of the plant extracts prepared in Example 2 in order to examine the antibacterial and antifungal activity according to the kind and concentration of organic acid. And 1.0% (W / V). Thereafter, the antimicrobial activity was measured in the same manner as in Test Example 1. The results are shown in Table 7.
As shown in Table 7, the growth of bacteria was significantly inhibited by the concentration of organic acid added rather than the type of organic acid added. In addition, lactic acid was found to have a higher effect on bacterial growth inhibition than other organic acids.
Example 3: Preparation of natural preservative
(W / V) 1,3-butanediol containing 0.5% (W / V) citric acid and 0.5% (W / V) lactic acid respectively was mixed with the plant extract prepared in Example 2 at a concentration of 5.0% To prepare a natural preservative.
Test Example 6: Measurement of antimicrobial activity of a natural preservative
The antimicrobial activity of the natural preservative prepared in Example 3 was measured in the same manner as in Test Example 1. For the control bacteria, commercially available natural preservatives, X, Y and Z, were used. The results are shown in Table 8.
As shown in Table 8, the natural preservative containing the mixture of the plant extract according to the present invention exhibited more excellent antibacterial and antifungal activity as compared with the antimicrobial activity of commercially available natural preservatives.
Claims (7)
Wherein the extract is extracted from a plant leaf, stem, core material, bark, flower or a combination thereof.
Wherein the extract is extracted with water, a C 1 to C 4 alcohol, ethyl acetate or a mixed solvent thereof.
Characterized in that the mixture is contained in an amount of 1% to 10% by weight, based on the total amount of the composition, of an antimicrobial and antifungal composition
Wherein the mixture is any one selected from the group consisting of antimicrobial and antifungal agents:
(1) a mixture comprising a 1: 1: 1 weight ratio of extracts of ground-up part, rabbit top part and phloem stalactite stem;
(2) a mixture comprising a 1: 1: 1 weight ratio of barkwood bark, foliar russell trunks and extracts of large crane flower flowers;
(3) a mixture comprising a 1: 1: 1: 1 weight ratio of a mane sprout leaf, a rabbit top part, an extract of a foliage of Sumatran spikelets and a large crane flower;
(4) a mixture comprising an extract of a 1: 1: 1: 1 weight ratio of a root lacquer tree stalk, a rabbit top part, a foliage lacquer tree stalk and a large crane flower flower;
(5) a mixture comprising a 1: 1: 1: 1 weight ratio of foliar russia leaf, rabbit top part, foliage of Sumac lacquer tree and extract of large crane flower;
(6) a mixture comprising a 1: 1: 1: 1 weight ratio of extracts of cassia bark, rabbit topsoil, foliage lacquer tree stems and large crane flower flowers;
(7) A mixture comprising extracts of 1: 1: 1: 1 weight ratio cadmium core, rabbit top, foliage lacquer tree stems and large crane flower flowers.
Wherein said composition is selected from the group consisting of Bacillus cereus , Enterobacter aerogenes , Listeria monocytogenes , Staphylococcus epidermidis ), Streptococcus mutans mutans , Escherichia coli , Proteus vulgaris), Salmonella Entebbe Rica (Salmonella enterica subsp. Enterica), Salmonella Entebbe utility disk (Salmonella enteritidis), Vibrio para Molly Tee hee Caicos (Vibrio parahaemolyticus ), Vibrio vulnificus , Candida albicans ( Candida albicans ), Debariomyces hansenii there is . hansenii , Pichia anomala , Aspergillus, flavus , Mucor circinelloides , Penicillium < RTI ID = 0.0 > digitatum and Rhizopus oryzae ), which has an antibacterial activity against a microorganism selected from the group consisting of:
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| KR1020150042684A KR20160115311A (en) | 2015-03-26 | 2015-03-26 | Antibacterial and antifungal composition comprising plant extracts |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20200031729A (en) * | 2018-09-14 | 2020-03-25 | 대한민국(환경부 국립생물자원관장) | Composition having Antimicrobial Effects Using Croton poilanei Extract |
| KR20200031730A (en) * | 2018-09-14 | 2020-03-25 | 대한민국(환경부 국립생물자원관장) | Composition having Antifungal Effects Using Croton poilanei Extract |
| KR20200063438A (en) | 2018-11-28 | 2020-06-05 | 주식회사 뉴랜드올네이처 | Cosmetic composition comprising Hosta longipes extract or fermented product and use thereof |
| KR20200069765A (en) * | 2018-12-07 | 2020-06-17 | 대한민국(농촌진흥청장) | Composition for controlling cobweb disease comprising extract of Hosta longissima HONDA as an active ingradient and uses thereof |
| KR20200092186A (en) * | 2019-01-24 | 2020-08-03 | 대구한의대학교산학협력단 | Composition comprising glycoprotein isolated from Rhus verniciflua stokes for preventing or treating sepsis or septic shock |
| KR20210071226A (en) * | 2019-12-06 | 2021-06-16 | 대한민국(농촌진흥청장) | Composition for controling plant diseases comprising Hosta longissima extracts and fractions, and uses thereof |
| KR102368892B1 (en) * | 2021-05-20 | 2022-03-03 | 주식회사 유진큐어 | A composition for anti-inflammatory comprising extracts of single leaf cremastra and sigesbeckia glabrescens makino |
| KR20220081111A (en) * | 2020-12-08 | 2022-06-15 | 대한민국(농촌진흥청장) | Composition for controling plant diseases comprising Hosta longissima extracts and fractions, and uses thereof |
-
2015
- 2015-03-26 KR KR1020150042684A patent/KR20160115311A/en not_active Application Discontinuation
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20200031729A (en) * | 2018-09-14 | 2020-03-25 | 대한민국(환경부 국립생물자원관장) | Composition having Antimicrobial Effects Using Croton poilanei Extract |
| KR20200031730A (en) * | 2018-09-14 | 2020-03-25 | 대한민국(환경부 국립생물자원관장) | Composition having Antifungal Effects Using Croton poilanei Extract |
| KR20200063438A (en) | 2018-11-28 | 2020-06-05 | 주식회사 뉴랜드올네이처 | Cosmetic composition comprising Hosta longipes extract or fermented product and use thereof |
| KR20200069765A (en) * | 2018-12-07 | 2020-06-17 | 대한민국(농촌진흥청장) | Composition for controlling cobweb disease comprising extract of Hosta longissima HONDA as an active ingradient and uses thereof |
| KR20200092186A (en) * | 2019-01-24 | 2020-08-03 | 대구한의대학교산학협력단 | Composition comprising glycoprotein isolated from Rhus verniciflua stokes for preventing or treating sepsis or septic shock |
| KR20210071226A (en) * | 2019-12-06 | 2021-06-16 | 대한민국(농촌진흥청장) | Composition for controling plant diseases comprising Hosta longissima extracts and fractions, and uses thereof |
| KR20220081111A (en) * | 2020-12-08 | 2022-06-15 | 대한민국(농촌진흥청장) | Composition for controling plant diseases comprising Hosta longissima extracts and fractions, and uses thereof |
| KR102368892B1 (en) * | 2021-05-20 | 2022-03-03 | 주식회사 유진큐어 | A composition for anti-inflammatory comprising extracts of single leaf cremastra and sigesbeckia glabrescens makino |
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|---|---|---|---|
| A201 | Request for examination | ||
| E902 | Notification of reason for refusal | ||
| E601 | Decision to refuse application |