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    • 专利摘要:
    PURPOSE: A biocidal agent containing a volatile extract of one or more natural materials as an effective component and a method for preservation of cultural assets using the agent are provided. The biocidal agent can be effectively used in preservation of cultural assets because it is excellent in durability of medicinal efficacy and free from toxicity. CONSTITUTION: The biocidal agent contains a volatile extract of one or more natural materials selected from the group consisting of Eugenia caryophyllata Thunberg, Boswellia carterii Birdwood and star anise, wherein the volatile extract of Star anise is 50% by volume or more, based on the total composition. Bacteria and insects that live in cultural assets are Trichoderma koningii, Mucor hiemalis, Aspergillus niger, Penicillium funiculosum as bacteria and Reticulitermes speratus, Lyctus linearis and Sitophilus oryzae L.
    公开号:KR20020063375A
    申请号:KR1020010003996
    申请日:2001-01-29
    公开日:2002-08-03
    发明作者:정용재；이규식
    申请人:대한민국 (관리부서:국립문화재연구소)；
    IPC主号:
    专利说明:

    Biocide composition comprising volatile extracts of natural medicines as effective components for conservation of cultural properties, and method for conserving cultural properties using the same
    [10] The present invention relates to a biocidal composition for the preservation of cultural properties using a volatile extract of natural products and a method for preserving the cultural property using the same, more specifically, the volatility of one or more natural products selected from the group consisting of cloves, frankincense and octagonal fennel The present invention relates to a biocide composition for preserving cultural property as an active ingredient and a method for preserving the cultural property using the same. In particular, it relates to a biocide composition having an antimicrobial and insecticidal effect on fungi and larvae inhabiting organic cultural properties, and a method for preserving cultural properties using the same.
    [11] It is the principle of nature that all materials on earth gradually change from high molecular material to low molecular material by the law of material circulation. Due to the principles of the natural world, cultural assets also appear in various forms of change over the years, which means the damage of cultural properties.
    [12] The damage factors involved in changing cultural properties can be numerous, but if they are largely divided into natural and artificial factors, they can be divided into physical, chemical and biological factors. Usually, cultural properties are damaged due to a complex action rather than a single action of the above factors.
    [13] When cultural properties are classified in terms of materials, they are divided into inorganic cultural properties and organic cultural properties. Mineral cultural properties generally consist of metal, stone, and glass, and organic cultural properties consist of wood, paper, fiber, and leather. In the case of small cultural assets, several kinds of materials are mixed, but in the case of wooden buildings, dozens of kinds of materials are usually mixed. Inorganic cultural properties are produced from inanimate objects and have a high melting point and boiling point, so they do not burn, and because they have little water content, they do not require moisture.They are generally weak in humidity, but hardly damage to bacteria and insects. Organic cultural properties, on the other hand, are made from living organisms and must contain certain moisture in order to maintain their shape. They are easily burned due to their low melting point and boiling point. The material properties of these organic cultural properties are provided as a nutrient source for fungi and insects, and the damages caused by erosion and erosion are almost impossible to recover from the circle. Therefore, the prevention of damage by biological factors is important in preserving organic cultural properties.
    [14] If damage caused by fungi or insects is in progress, the cultural property must be killed immediately. Various insecticides and fungicides are used to prevent damage to organic cultural properties by these insects and microorganisms. In general, the development of pesticides and disinfectants has been concentrated on the development of pesticides to solve the food problems of mankind through the quantitative increase of agricultural products, some of which have been applied to the preservation of cultural property. Gas fumigation using a mixed gas of ethylene oxide and methyl bromide, which is used for quarantine of soil herbicides, pesticides and agricultural products in agriculture, is a representative method of killing cultural properties that are being damaged by fungi and insects. It is used for controlling bacteria and insects.
    [15] As insecticides, since DDT became a practical insecticide in 1938, organic chlorine-based insecticides such as BHC, aldrin, and endrin began to be developed. The use of synthetic insecticides has exploded and the development of agricultural products and the improvement of human health have also been remarkable. Since the excellent effects of DDT have been demonstrated, an increasing number of pesticides have been developed and marketed, including organochlorine, organophosphorus and carbamite pesticides (Metcalf, RE 1980. "Changing role of insecticides in crop protection" Ann. Rev. Entomol. 25: 219-256.). It was true that the development of such insecticides had a great effect on food production and health improvement, but side effects such as the emergence of resistant pests, the major pests of latent pests, harmful to the human body and various environmental pollution problems began to appear.健, 桐 俗 圭 治, 金 澤 1973. 生態 系 ど 農藥. 岩 波 書店, 東京 .214: Brown, AWA 1978. Ecology of Pesticides.p. 525. John Wiley & Sons, New York; 金 澤 純. 田中 二 良. 1979. 水 生生 ど 農藥. 東京. 208.
    [16] As such, organic synthetic insecticides and disinfectants have contributed greatly to the vitalization of agricultural work, but on the other hand, environmental pollution and residual toxicity of the human body have emerged as practical problems. Accordingly, the development of insecticides and fungicides in natural products existing in nature is known as the most preferable way to replace the problems caused by organic synthetic pesticides.
    [17] It is known that there are about 300,000 kinds of plants on earth and many secondary metabolite components are distributed in these plants. Animals and microorganisms excrete the end metabolites out of the body, but since most of the plants accumulate in the body, it is no exaggeration to say that all of the secondary metabolites are plant components. Of these plant components, humans have discovered about 10% of the substances so far, and only a few of them are used in pharmacology, dyes, and beverages. Moreover, even if the substance is known to be currently identified and used, there are also a number of other activities that the substance has not been found.
    [18] In particular, research on the identification of antimicrobial active substances and the identification of active bodies from herbal and edible plant extracts that have been indirectly innocuous and has been used as a disease treatment agent or food material for a long time has been actively conducted.
    [19] Representative plant extracts include spices and herbal medicines, the oldest of which are garlic, onions, peppers, and many other spices, and many herbals such as ganoderma lucidum and licorice, and extracts of edible plants including mugwort and green tea have antibacterial properties. It is reported to be. Allicin in garlic is an antimicrobial active that acts as an inhibitor of -SH enzyme (Park, UY, Chang, DS and Cho, HR 1992. Antimicrobial effect of Lithospermum erythrorhizon extract.J. Korea Soc.Food Nutr 21: 97-100.), And antimicrobial activity against Pythium ultimum , which causes crop stalks from medicinal plants, has been reported to be effective in 9 species, including charcoal, cyglia and rhubarb. (Bae Su-bong, Yeon-sun Oh. 1990. Exploration of antimicrobial medicinal plants for the control of Pythium ulimum soil pathogen. Korean Journal of Mycology 18: 102).
    [20] Essential oils are primarily secondary metabolites secreted by the endocrine glands of plants. They are found in almost all parts of the plant, including flowers, leaves, fruits, wood, and bark.
    [21] In the late 18th century, significant advances in chemical studies began to accelerate the study of chemical molecular structures of essential oils, and many kinds of essential oils derived from natural resources have been known in modern times. The basic skeleton is a component of C 10 -C 15 which has a wide variety of structures and functional groups, and is used as a natural fragrance resource in the East and West. In other words, natural essential oils with unique fragrances and flavors are applied in a surprisingly wide range of industries, including perfumes, flavors and pharmaceuticals (Seoul National University Natural Products Research Institute. Research on the development of traditional natural flavors.
    [22] In the field of preservation of cultural properties using natural products, the results of insect repellent effects on medicinal plants used as insect repellents from ancient times reported that there was an insect repellent effect on clove and cinnamon. Studies on the Insect Repellent Effectiveness Ph.D degree of Dissertation. Basic experiments and clothing including Aureus ( S. aureus ) reported more than 82% antimicrobial activity (Lee Hyun Sook. 1998. Dyeing, antibacterial, deodorant, master degree of Dissertation, Graduate School of Sungkyunkwan University.) Although there have been few studies on insecticides and bactericides for the preservation of tributaries and textile cultural assets due to fungi and insect damage using natural products, few studies have been conducted.
    [23] The currently widely used method of killing fungi and worms in cultural properties is gas fumigation. At present, a mixed gas of ethylene oxide and insecticide methyl bromide is used for controlling fungi and insects that cause cultural properties worldwide. However, the gas fumigant is excellent in stability and fast-acting effect, but since there is no residual oil, it is preferable to carry out the treatment of the dry wood after the fumigation by a separate preservative. In addition, methyl bromide, a colorless and odorless toxic chemical currently used, is 50 times more potent to control the pollution of the ozone layer than CFC, and developed countries have stopped using it except quarantine.
    [24] Fumigation treatment is a very excellent means of killing the perpetrated organisms. However, since the drug is in gaseous state, it does not remain in the material of the cultural property, so it has little effect on the material. Although there is an advantage, the drug does not last for a long time and has a disadvantage of being very harmful to the human body. Therefore, in the case of small artifacts it is necessary to install a low-toxic and persistent insect repellent or fungicide to prevent the damage of the fungus.
    [25] Currently, 15-20 g / m3 of p-dichlorobenzene is used as an insect repellent, 20g / m3 of paraformaldehyde (p-formaldehyde) and 60g / m3 of thymol are used as a chemical, but it is applied to a cultural property as a chemical. There is a problem below.
    [26] In particular, organic cultural properties deteriorate naturally over a long period of time, and the material is very weak. In addition, due to its material properties, discoloration and hydrolysis may occur depending on the conditions of the surrounding environment (light, temperature, humidity, dust, harmful gas, etc.), which may accelerate deterioration. It should be selected and used that does not affect.
    [27] Therefore, the inventors of the present invention solve the persistence and toxicity of the drug, which is a problem of the prior art for the preservation of the cultural property, as a result of earnest research to search for pesticides and fungicides for the preservation of the fiber and tributary cultural property from natural products, the ancient Non-volatile Fragrances of Natural Herbs and Spices Used in Korea from the Five Incenses added to Buddha's Dress Relics, Cloves, Frankincense, Gwakyang, Celadon, Aloe, and Octagonal Fennel And extracting volatiles to confirm biocides, and analyzing the fragrance components of the volatiles to determine the extent to which they affect the material properties of organic cultural properties. Eventually, the biocide composition including volatile substances of one or more natural products selected from the group consisting of cloves, frankincense and octagonal fennel was found to be excellent for preserving cultural property, and thus, the present invention was completed.
    [28] Accordingly, an object of the present invention is to provide a biocide composition effective for controlling fungi and worms inhabiting cultural assets, especially tributaries and fibrous cultural assets.
    [29] Still another object of the present invention is to provide a method of preserving cultural property using the biocide composition.
    [1] 1 shows fungi (A: Mucor hiemalis ), B: Aspergillus niger , C after 72 hours using volatile extract extracted by solvent-distillation extraction from cloves. : It is a figure which shows the growth inhibitory effect of Penicillium funiulosum , D: Trichoderma koningii .
    [2] Fig. 2 shows fungi (A: mucor hyemalis, B: aspergillus niger, C: penicillium funniolsum) after 72 hours using volatile extract extracted from solvent frankincense by solvent-distillation extraction method. , D: A diagram showing the growth inhibitory effect of Trichoderma Koninji.
    [3] Figure 3 shows fungi (A: mucor hyemalis, B: Aspergillus niger, C: penicillium funniolo) after 72 hours using volatile extract extracted from octagonal fennel by solvent-distillation extraction method. It is a figure which shows the growth inhibitory effect of breath, D: Tricorderma Koninjieye).
    [4] Figure 4 shows fungi (A: mucor hyemalis, B: aspergillus niger, C: penicillium funniolsum, D: trichoderma conin, 72 hours after using the control group (methylene chloride). It is a figure which shows the growth inhibitory effect of GIA.
    [5] Figure 5a is a mixture of clove and octagonal volatile extracts (A: 1: 1, B: 1: 2, C: 1: 3, D: 1: 4) using a mixture (total volume 10ul) It is a figure which shows the growth inhibitory effect on Aspergillus niger after 72 hours.
    [6] Fig. 5B shows the volatile extract of clove (A), octagonal fennel (C), and a mixture (B) containing clove and volatile extract of octagonal fennel at a volume ratio of 1: 2 in a total volume of 10 μl, after 72 hours. Figures comparing the growth inhibitory effect on Pergillus niger.
    [7] Figure 6a is a diagram showing the insecticidal ability 48 hours after the termite inoculation in the volatile extract (left) and the control (right) of the clove.
    [8] Figure 6b is a view showing the insecticidal 24 hours after inoculation of termites in the volatile extract (left) and the control (right) of the octagonal fennel.
    [9] 7 is a view showing the insecticidal ability 7 days after inoculation of rice weevil (A) and thigh wood (B) in the volatile extract of clove, frankincense and octagonal fennel respectively.
    [30] The biocide composition for preserving cultural properties as a volatile extract of the herbal medicine according to the present invention comprises a volatile extract extracted from at least one natural product selected from the group consisting of cloves, frankincense and octagonal fennel.
    [31] In addition, the biocide composition for preserving the cultural property contains at least volatile extract of octagonal fennel, preferably characterized in that 50% or more based on the total volume of the composition.
    [32] As used herein, the term "volatile extract" means a material obtained by a solvent-distilled extraction method.
    [33] Thus, the biocide composition contains a fragrance component of at least one natural substance selected from the group consisting of cloves, frankincense and octagonal fennel, wherein the fragrance component of the natural substance contains eugenol or anetol.
    [34] Clove ( Eugenia caryophyllata THUNBERG), which is a component of the biocide composition of the present invention, is an unflowered bud of Eugenia caryophyllata Thunberg, an evergreen tree belonging to Myrtaceae.
    [35] Frankincense ( Boswellia carterii BIRDWOOD), which is a component of the biocide composition of the present invention, is native to the Middle East and India, and belongs to the olive family and is excellent in aromaticity. In the present invention, as well as excellent antimicrobial activity against the experimental strain is added for the aromaticity.
    [36] The octagonal fennel ( Star anise or Illicium verum Hook.fil.), Which is one component of the biocide composition of the present invention, is grown in southwestern China (Guangxi) and is also grown in North Vietnam, southern India, Japan, and the like, and has a height of 6-10 m. A tree belonging to the family Magnoliaceae.
    [37] The biocide composition of the present invention is preferably characterized in that it is applied to organic cultural properties, such as cultural properties, in particular fibers, paper, wood, leather.
    [38] The most common fungi inhabiting these cultural properties are known to be filamentous fungi, and currently known filamentous fungi (about 4,320 genus and 46,300 species) are basidiomycetes (about 550 genus and 15,000 species), junctional fungi (about 245 genus and 1,300 species) It is classified into fungi (about 1,700 genus, 15,000 species) and incomplete fungi (about 1,825 genus, 15,000 species). In particular, there are about 100 species of filamentous fungi that occur in organic cultural properties, including the species of Chatomium sp., Trichoderma sp., Aspergillus sp., And Penicillium sp. sp.), Alternaria sp., Stachybotrys sp., etc. Also known as fungi and discoloration bacteria that cause wood decay and contamination. Pergillus species, Penicillium species, Fusarium sp., Mucor sp., Rizopus sp.
    [39] In the present invention, in order to test the antimicrobial activity of the biocide composition, Mukor Hiemalis (Zygomycotina) (Mucor hiemalis, Accession number: ATCC 8690), Trichoderma Koninjiai (Deuteromycotina)Trichoderma koningii, Accession number: KCTC 6042) and Ascomylota niger (Ascomycota)Aspergillus niger, Accession number: KCCM 11239) and penicillium funniolum (Penicillium funiulosum, Accession number: KCCM 12040).
    [40] Representative larvae inhabiting organic cultural properties also include Coleoptera, Termite (Isoptera), Thysanura, Psocoptera, Hymenoptera, and Blattaria. Termite damage is most severe.
    [41] In the present invention, in order to confirm the insecticidal properties of the biocide composition, termites of the termite ( Reticulitermes speratus ), the bark of the bark ( Lyctus linearis GOEZE) and rice weevil ( Sitophilus oryzae L. Use
    [42] The termites are small insects, most of which are less than 1 cm in length, and feed on plant materials such as wood and dead plants. Termites inflict cultural assets such as wood, tributaries, and fibres, and the distributed species in Korea mainly damage the wood of houses and wood conditioners.
    [43] The thigh wood adult insects escaped by drilling holes of about 1 to 2 mm on the surface of the wood to avoid 3 to 8 mm. The damage of wood by the thighwood roots occurs mainly in the larval stage, and is usually cooled by making tunnels along the fiber direction in sapwood.
    [44] The length of the rice weevil adult is about 2.3-3.4mm, reddish brown, many entangled traces on the back, and the larva is about 2.5-3.0mm in the shape of white maggot. It occurs 3 to 4 times a year and overwinters as a larva or adult, and females eat grains from the end of May. Adults add dozens of grains. It is based on all kinds of cereals and processed foods, and is used as a test insect to check the pesticidal properties of chemical pesticides.
    [45] In the present invention, in order to assay the insecticidal and antibacterial activity against the bacteria and worms, first, non-volatile substances and volatile substances are extracted from natural products. Preferably, as a component for use as a biocide composition for preserving cultural property, a volatile substance of clove, frankincense and octagonal is selected from five flavors (clove, frankincense, quill, celestial aroma, and fennel).
    [46] In addition, the volatile extract may be used singly or selectively, depending on the kind of the main fungi, worms inhabiting the cultural property.
    [47] As a method for extracting nonvolatile and volatile substances of these natural products, first, nonvolatile materials are obtained by using ultrasonic extraction, and volatiles are extracted by using solvent-distillation extraction.
    [48] The biocide composition of the present invention for application to a cultural property, preferably organic cultural properties such as fibers, tributaries can be used by diluting the extracted volatile extract pure or in a concentration suitable for use.
    [49] Means for applying the biocide composition to the cultural property include, but are not limited to, a direct spray method, application method, fumigation method, aroma treatment method, packaging with the biocide composition is coated.
    [50] In addition, the formulation for applying the biocide composition of the present invention to cultural properties include aerosols for direct spraying, tinctures for fumigating or aroma treatment, liquids, matrix gels for sustained release formulations, and the like. It is not limited.
    [51] The effective amount of the cultural property applied to the biocide composition of the present invention varies depending on the components and the application means, but for example, in the aroma treatment method, the concentration in the air is usually adjusted to 50 ml / m 3 or more, preferably 125 ml / m 3 or more. .
    [52] Hereinafter, although an Example, a comparative example, and a test example demonstrate this invention in detail, this invention is not limited to these examples.
    [53] Example
    [54] 1. Experimental Materials
    [55] ① Herbal Medicine
    [56] Cloves ( Eugenia caryophyllata THUNBERG), frankincense ( Boswellia carterii BIRDWOOD), and octagonal fennel ( Ilicium verum Hook.fil.) Were purchased and used for various experiments.
    [57] ② Experiment bacteria
    [58] Experimental strains for antimicrobial activity were identified from Mucor Hiemalis (ZCC: ATCC 8690), a conjugated fungus (Zygomycotina), and Trichoderma Koninzieye (KCTC 6042), an incomplete fungus (Deuteromycotina), and Korea Microorganism Conservation Center. Ascomycota Aspergillus niger (Accession No .: KCCM 11239) and Penicillium funniusum (Accession No .: KCCM 12040) were distributed in total and four kinds of potato glucose agar medium at 26 ° C incubation temperature. (Potato dextrose agar, manufactured by Dipco Co., Ltd.) was used. As insects to check insecticidal properties, broad-leafed bark, termite and rice weevil were used.
    [59] Example 1 Extraction of Volatile Compounds from Herbs
    [60] 100 g of herbal medicines of clove, frankincense and octagonal extract were extracted for 3 hours using 125 ml of methylene chloride re-distilled in a solvent-distilled extraction apparatus. Anhydrous sodium sulfate was added to the extract to remove water, and concentrated to 10 ml each under a nitrogen gas stream, which was used for antimicrobial and insecticidal experiments, and was used as an analytical sample for GC / MS.
    [61] Example 2 Preparation of a Mixture of Volatile Extracts of Herbs
    [62] The clove volatile extract prepared in Example 1 and the octagonal volatile extract were mixed in a volume ratio of 1: 2 and used for the first antimicrobial activity test.
    [63] Comparative Example 1: Extraction of Non-Volatile Compounds from Medicinal Herbs
    [64] 4 g of herbal medicines of clove, frankincense and octagonal were weighed and transferred to a 50 ml Erlenmeyer flask, and ultrasonic extraction was performed at room temperature with 20 ml of ethanol for 1 hour. The extract was collected by filtration with filter paper, and 20 ml ethanol was added to the residue, followed by repeated ultrasonication for 1 hour, followed by filtration. The collected extract was concentrated under reduced pressure at 40 ° C. with a rotary evaporator, and dissolved in 4 ml of ethanol, which was used for the first antibacterial and insecticidal experiment.
    [65] Test Example 1 Antibacterial Activity Measurement
    [66] 1) Antimicrobial Activity of Volatile Extracts
    [67] Each sterilized growth medium was coagulated by dispensing 15 ml each of Petri dishes (Φ = 80 mm, h = 10 mm), and the sterilization medium was dispensed by dispensing 5 ml of each medium into test tubes. Subsequently, various test bacteria liquids pre-incubated while storing in a 50 ° C. water bath were aseptically added and mixed well, and then dispensed onto a base medium to make a double plate medium.
    [68] The volatile extract obtained in Example 1 was absorbed by 50 µl into sterile paper disks, and then placed on a Petri dish lid so as not to be directly closed on a test plate medium. Subsequently, the resultant was sealed with a sealing tape, incubated at 26 ° C. for 3 days, and then the clear zone (mm) of the upper part of the disk was measured to search for antibacterial activity. The results are shown in the table below and FIGS.
    [69] Growth inhibition zone (mm) clovesfrankincenseOctagonal fennel Mukor Hiemalis654080 *Aspergillus Niger5880 * 80 *Penicillium funniumsum80 * 80 * 80 *Trikoderma Koninjiai57-80 *
    [70] 80 * means 100% growth inhibition rate.
    [71] In order to exclude the antimicrobial activity of the solvent itself, only methylene chloride was added and used as a control (FIG. 4).
    [72] As shown in the table and FIGS. 1 to 3, the volatile extracts of the cloves formed a clear zone in the Mucor hyemalis, Aspergillus niger, Trichoderma Koninziai experimental strains and penicillium funniolsum This inoculated plate medium was able to observe that the growth of bacteria was suppressed on the entire surface (FIG. 1). Volatile extract of frankincense is penicillium funniolsum, except Trichoderma Koninji,Strong antimicrobial activity was shown on the entire surface of plate media inoculated with Mucor Hiemalis and Aspergillus niger (FIG. 2). Volatile extract of octagonal fennel inhibited the growth of bacteria inoculated on plate medium in all strains showed the strongest antibacterial activity among the experimental medicine (Fig. 3).
    [73] 2) Antimicrobial activity against mixtures of volatile extracts
    [74] First, in order to confirm the minimum inhibitory concentrations of the four strains of the volatile extracts of the clove and octagonal fennel, each of the volatile extracts in 50 Petri dishes (Φ = 80mm, h = 10mm) inoculated with 50 ㎕ and 25 ㎕, respectively. , 10µl, 5µl and 2µL were absorbed into the paper disk and placed in the cap so as not to directly contact the medium. After 3 days of incubation, the antimicrobial activity was measured by measuring the clear zone (mm). The results are shown in the table below.
    [75] Cloves in mmOctagonal Fennel (unit: mm) 50 μl25 μl10 μl5 μl2 μl50 μl25 μl10 μl5 μl2 μl Aspergillus Niger585245402580 * 28--- Penicillium funniumsum80 * 80 * 80 * 80 * 6080 * 80 * --- Mukor Hiemalis656042402480 * 72--- Trikoderma Koninjiai575552442880 * 80 * ---
    [76] 80 * means 100% inhibition.
    [77] As can be seen from the table, the volatile extract of the clove showed antimicrobial activity even at low concentration (below 5μl), but showed antibacterial activity against all strains at 10μl or more. The volatile extract of octagonal fennel showed no antibacterial activity at concentrations of 10 μl or less, but showed antimicrobial activity at concentrations of 25 μl or more, so the minimum inhibitory concentrations of the volatile extracts of clove and octagonal fennel were measured at 10 μl and 25 μl, respectively.
    [78] Antimicrobial activity was measured in the same manner as in 1) of Test Example 1, except that the volatile extracts of the clove and octagonal fennel obtained in Example 2 were mixed and used in a volume ratio of 1: 1 to 1: 4. As the test bacteria, Aspergillus niger with the smallest growth inhibitory effect was used in the above table.
    [79] As shown in FIG. 5A, the mixture of the volatile extracts (total volume of 25 μl and 10 μl, respectively) observed that the growth of bacteria was suppressed on the entire surface in both plate media inoculated with Aspergillus niger. Could. The results are shown in the table below.
    [80] Mixture (volume ratio)Aspergillus Niger Clove: Octagonal fennel25 μl10 μl 1: 15028 1: 25250 1: 35330 1: 45212
    [81] As can be seen from the table, it can be seen that high antibacterial activity was exhibited even at the minimum concentration of volatile extracts of cloves and octagonal fennel. In particular, the mixture of clove and octagonal volatile extract 1: 2 showed the highest antimicrobial activity, and as shown in FIG. 5B, their antimicrobial activity (clear zone 50mm) and clove (clear zone 45mm) and octagonal fennel (clear Zone 0 mm) showed excellent antimicrobial activity at a concentration of 10 μl compared to each volatile extract. Overall observations with the clear zone and the naked eye show a synergistic effect of about 2 times.
    [82] 3) Evaluation of Effective Amount for Antimicrobial Activity of Volatile Extracts
    [83] Airtight container (2000 cm 3) to check the antibacterial activity of the volatile extract of clove and octagonal fennel obtained in Example 1 and the mixture of clove and octagonal fennel prepared in Example 2 (volume ratio 1: 2) according to the appropriate concentration in air. After preparing PDA medium, inoculated with 300 μl (1 × 10 4 spores / ml) of the test strain Aspergillus niger, and 100 μl, 250 μl, 500 μl and 1000 μl of each extract were absorbed into the filter paper. The growth rate of the strains was compared after culturing for 3 days by placing the cap on the lid so as not to directly contact the medium. The results are shown in the table below.
    [84] Aspergillus Niger 100 μl250 μl500 μl1000 μl Cloves-50%60%90% Octagonal fennel-50%80%100% Mixture (octagonal fennel 2: cloves 1)60%90%100%200%
    [85] As can be seen from the table, the clove volatile extract showed a growth inhibition of 90% at 1000μl concentration, the octagonal fennel showed a growth inhibition of more than 80% at 500μl. However, the mixture of clove and octagonal volatile extract (volume ratio 1: 2) showed growth inhibition of more than 90% at 250 μl concentration and showed more than 60% growth inhibition at 100 μl concentration, and the mixture showed strong sterilization power at the concentration of 125 ml / ㎥ or more. It can be seen.
    [86] Comparative Test Example 1 Determination of Antimicrobial Activity of Nonvolatile Extracts
    [87] Each sterilized growth medium was coagulated by dispensing 15 ml each of Petri dishes (Φ = 80 mm, h = 10 mm), and the sterilization medium was dispensed by dispensing 5 ml of each medium into test tubes. Subsequently, various test bacteria liquids pre-incubated while storing in a 50 ° C. water bath were aseptically added and mixed well, and then dispensed onto a base medium to make a double plate medium.
    [88] The nonvolatile extract obtained in Comparative Example 1 was absorbed by 50 µl in sterile paper discs, placed on a test plate medium, and then incubated at 26 ° C. for 3 days, followed by a clear zone (mm) around the disc. Was measured to search for antimicrobial activity.
    [89] In order to exclude the antimicrobial activity of the solvent itself, only the same ethanol as the treatment concentration was used as a control.
    [90] The results of measuring the antimicrobial activity of the nonvolatile extracts are shown in the following table.
    [91] Growth inhibition zone (mm) clovesfrankincenseOctagonal fennel Mukor Hiemalis34-- Aspergillus Niger44-- Penicillium funniumsum40-6 Trikoderma Koninjiai40--
    [92] As can be seen from the table, was observed a clear zone for all test strains in the non-volatile extract of clove, had a non-volatile extract from frankincense, did not show the antimicrobial activity in all strains, non-volatile extract of octagonal fennel is penny chamber It has a weak antimicrobial activity in Leeum Funniulsum.
    [93] As can be seen from the above figures and tables, it can be seen that the volatile extracts and their mixtures have an antimicrobial effect on the main fungus inhabiting cultural properties compared to the nonvolatile extracts, and also compared to the volatile extracts. It was found that the mixture had a synergistic effect.
    [94] Test Example 2: Determination of Insecticidality of Volatile Substances
    [95] 1) Insecticide against termites
    [96] 400 μl of each extract obtained in Example 1 was absorbed into a paper disk and placed on a filter paper (5 × 5 cm, 1.0 g). In the same population, 10 soldiers and 100 workers were collected and placed in a sealed box (2000cm 3 ) controlled with humidity by gypsum and sealed.
    [97] 2) Insecticidal properties on the thigh wood
    [98] Twenty thigh wood chips were placed in Petri dishes (Φ = 80 mm, h = 10 mm) filled with 4 g of sawdust and absorbed 200 μl of each extract obtained in Example 1 on a paper disk. Thereafter, direct contact with food from insects was placed and sealed to observe mortality after 14 days of incubation in a 30 ° C thermostat.
    [99] 3) Insecticide against rice weevil
    [100] 20 rice weevil was put in a petri dish (Φ = 80mm, h = 10mm) filled with 10g of brown rice and treated the extract in the same manner as 2) and then cultured for 14 days at 30 ℃ constant temperature and confirmed the mortality.
    [101] At this time, in order to exclude the insecticide of the solvent itself added, all tests were set to the control group added only methylene chloride in the same concentration as the treatment concentration.
    [102] The results of confirming the insect killing by culturing after placing the volatiles on the thigh wood, termites and rice weevil not directly close to the volatiles are shown in the following table.
    [103] activation clovesfrankincenseOctagonal fennel termite+++-+++ Thigh tree+-+++ Rice weevil--++
    [104] As can be seen from the table and FIG. 6, the insecticidal activity of termite octagonal volatile extract was within 24 hours after inoculation, and the volatile extract of cloves showed 100% mortality after 48 hours.
    [105] Insecticidal properties of the thigh bark showed volatile extracts of octagonal fennel and cloves 100% and 20% mortality, respectively, within 7 days after inoculation (FIG. 7).
    [106] The insecticidal activity of rice weevil was low, but the rice weevil was strong enough to be used as a test insect to confirm the pesticidal properties of chemical pesticides. Although difficult to follow, the volatile extract of octagonal fennel showed 20% mortality within 7 days after inoculation and 50% after 20 days).
    [107] Comparative Test Example 2: Determination of Insecticidality of Nonvolatile Extract
    [108] 1) Insecticide against termites
    [109] 1 g of filter paper was absorbed by 1 ml of each extract obtained in Comparative Example 1. Thereafter, 10 soldiers and 100 worker ants were collected from the same group, and fed to a humidity-controlled box with a gypsum and bred at 26 ° C. for 14 days to confirm mortality.
    [110] At this time, only ethanol was added at the same concentration as the treatment group as a control.
    [111] In order to confirm the insecticidal properties of the nonvolatile extract, the results of confirming the insecticidal properties of the termites against the ethanol extract are shown in the following table.
    [112] Number of termites that die in two weeks ControlclovesfrankincenseOctagonal fennel 1 day110110110110 2 days110-110110 3 days110-110110 9th110-90110 12 days100-60100 14 days90-5090
    [113] As shown in the table, it was confirmed that the ethanol extract of the clove was 100% mortality 48 hours after inoculation, the ethanol extract of frankincense and octagonal selves showed almost no mortality against termites.
    [114] As can be seen from the above figures and tables, it can be seen that the volatile extract has a pesticidal effect on the main larvae inhabiting the cultural property as compared to the nonvolatile extract.
    [115] Test Example 3: Component Analysis and Bioactive Substance Search
    [116] For the component analysis of each herbal extract obtained in Example 1 was analyzed by gas chromatography (GC) and gas chromatography / mass spectrometer (GC / MS). The instrument used Autospec (manufactured by Micromass, UK) and the column (DB-5) used a fused silica capillary column (30 m × 0.25 nm). The column temperature was programmed at 15 ° C./min up to 60-280 ° C. and the detector and injector temperatures were 290 ° C.
    [117] Bioactive substances were searched through TradiMed (Dongyang Media), a traditional oriental medicine database at Seoul National University.
    [118] The results of component analysis on the volatile extracts of cloves, frankincense and octagonal fennel are shown in the table below.
    [119] <Contents of Volatile Extract of Clove>
    [120] Ingredient numberRTcompound One13:43Benzoic acid, 2-hydroxy-, methyl methyl ester 216:12Cavicol 320:21Eugenol 421:42Trans-cayophilene 522:42α-fumulene 626:40Cayophilene oxide 727:38β-Eudesmol 828:10Cayophila-4 (12), 8 (13) -diene-5-β-ol 929:07Cayophilene oxide 1031:41Benzylbenzoate
    [121] <Vegetable Volatile Extract-Containing Ingredients>
    [122] Ingredient numberRTcompound One5:233-karen 28:101,8-cineol 38:47Trans-β-oxymen 49:39N-octanol 510:33L-Linarul 613:55Cyclopropane, 2- (1,1-dimethyl-2-pentenyl) -1,1-dimethyl-cyclopropane 714:36n-octyl acetate 816:561-bornyl acetate 926:29Hexyl octanoate 1036:01Manul 1136:49Neosembre A 1237:47Sembren-C 1338:00Bertisiol 1441:041,5,9-cyclotetedecatene, 1,5,9-trimethyl-12-
    [123] <Volatile ingredients containing octagonal fennel>
    [124] Ingredient numberRTcompound One5:25α-Pipen 27:34δ-3-karen 38:131-limonene 410:07α-terpinene 510:33L-Linarul 613:40α-terpineol 713:57Estragol 817:50Anetol 920:21p-methoxyphenylacetone 1020:02Methyl-4-methoxybenzoate 1122:25Trans-α-bergamoten 1222:431-propanone, 1- (4-methoxyphenyl)- 1323:56Trans-methyl isoeugenol 1425:57Nerolidol 1526:20Spaturenol 1626:332- (1-cyclopentenyl) furan 1728:18Crotonic acid, O-benzaldehyde doceter 1828:34Alpha-cardinol 1929:17Pheniculin 2043:411-hexene, 2- (P-anisyl) -5-methyl-
    [125] As can be seen from the table, there were 10 volatile components in the volatile extract of clove, 14 volatile components in the volatile extract of frankincense, and 20 volatile components in the volatile extract of octagonal fennel.
    [126] The chemical structural formulas of eugenol and anetol, the main components of the clove and octagon, are as follows:
    [127]
    [128] Eugenol
    [129]
    [130] Anethole
    [131] Test Example 4: Material stability test
    [132] 1) Experimental material
    [133] To determine the degree of damage to cultural property tributaries printing and two acquaintances after a preservation copy paper purchased in Insa-dong branch that kind (pH 7.0 or higher triglyceride basis weight 70~72.89g / m 3 (A4 size), the subsidiary Hansol first) Newspaper paper (basis weight 54g / m 3 , manufactured by Hansol Paper Co., Ltd.) was used and used in various experiments.
    [134] In addition, cotton yarn (100% cotton), silk yarn (100% silk, twisted Z) and embroidery thread (viscose rayon 100%) were purchased and used in various experiments to measure the degree of damage to the textile cultural properties.
    [135] 2) Experiment Method
    [136] ① Manufacture of paper and fiber specimens
    [137] A total of 45 pieces (3 types of paper × 15 pieces) of 15 pieces of 15 pieces of paper and 3 pieces of paper were 15 mm long and 110 mm long. X 30 specimens).
    [138] ② pretreatment of specimen
    [139] Filter paper which absorbed 0.5ml of volatile extracts of clove, frankincense and octagon in a sealed container in which 240 g of ammonium nitrate (NH 4 NO 3 ) was dissolved in 100 ml of distilled water and maintained at 60% humidity. 2g) was put together so as not to directly touch the paper and the fiber specimens, and maintained at a temperature of 80 ℃ and the samples were taken according to each condition and the material condition was measured.
    [140] For comparison, paper specimens and fiber specimens treated with methylene chloride (CH 2 Cl 2 ) only under the same conditions were used as controls.
    [141] ③ Material Condition Survey
    [142] end. Whiteness
    [143] In order to observe the color change of the paper material, it was measured using a colorimeter (Minolta CR-200, Japan), and the change in color was expressed by the color difference (ΔE) of the L * a * b * color system according to KS A 0063. . L * a * b * One of the perceptually even coverage of color space recommended in 1976 by the Commission International de I'Eclairage. CIE 1976 (L * a * b * ) is called the color space and is represented by CIELAB. The color difference (ΔE * ab ) calculation by L * a * b * colorimetric system is as follows:
    [144] ΔE * ab = ((ΔL) 2 + (Δa * ) 2 + (Δb * ) 2 ) 1/2
    [145] In the above formula, L * , a * , b * are the differences between the brightness indexes L * , chromatactic indexes a *, and b * of two objects in the L * a * b * color system defined in KS A 0067 respectively.
    [146] I. Acidity
    [147] The acidity of the material was measured to determine the degree of damage of the paper material. Acidity of the paper surface was measured using a pH meter (Hanna Hi 9024C, USA) in accordance with KS M 7053, and the averages were expressed.
    [148] All. Strength
    [149] In order to determine the physical state due to hardening and relaxation of paper materials and to measure the state of the material and the degree of aging, the strength was measured. The stiffness strength was measured using a stiffness strength device (Toyo Boseki MIT-S, Japan) under a 500g load in accordance with KS M 7065, and the results were averaged and displayed. The test pieces were cut into a width of 15 mm and a length of 110 mm to measure 15 sheets each.
    [150] la. Tensile strength
    [151] In order to measure the condition of the fibers and the degree of damage, the tensile strength of the fibers was measured. The actual tensile strength was measured with a real tensile strength tester (Daekyung Tech, Korea) based on KS K 0323, and the average of the results was displayed. The test piece was cut out to 30 cm in length, and each measured 30 pieces. The formula for tensile strength is as follows:
    [152] Tensile Strength (g / den) = S D / D
    [153] Where S D is tensile strength when dry and D is quantitative fineness.
    [154] hemp. Ion Chromatography
    [155] The change of anion was measured by ion chromatography (Dionex CD20, IP20, USA) in order to measure the chemical composition change of paper material by the fragrance component of volatile extract. Volatile extracts of clove, frankincense and octagonal fennel were placed in 5 50ml Erlenmeyer flasks containing 0.2g of three paper specimens pretreated at 80 ° C for 30 days. Ions in the paper were eluted. In order to remove the suspended paper fibers in each eluted solution, 1.2 ml was dispensed in an Eppendorf tube, and then centrifuged at 10,000 rpm for 10 minutes (Juan MR1822, USA), and the supernatant was collected to measure anion concentration. Are averaged and displayed respectively.
    [156] Measuring conditions
    [157] Column: AS4A-SC
    [158] Buffer: 1.8 mM Na 2 CO 3 + 1.7 mM NaHCO 3
    [159] Flow rate: 2ml / min
    [160] 3) Experiment result
    [161] ① Material stability result for feeder
    [162] end. Whiteness
    [163] In order to confirm the color change of the paper by the fragrance component of the volatile extract quality, the results of measuring the whiteness of two kinds of pre-treated preservative paper, newspaper paper, and Korean paper are shown in the following table.
    [164] Preservation 2NewspaperAfter Control11.6418.9319.72 cloves12.2819.4120.21 frankincense11.3218.0918.88 Octagonal fennel10.8918.2220.21
    [165] As can be seen from the table, the whiteness of the paper specimens by the volatile extract of clove, frankincense and octagonal fennel did not show a significant difference compared to the control.
    [166] I. Acidity
    [167] In order to confirm the pH change of the paper by the fragrance component of the volatile extract quality, the results of measuring the pH of two kinds of pretreated preservative paper, newspaper paper, and Korean paper are shown in the following table.
    [168] Preservation 2NewspaperAfter Control7.506.936.59 cloves7.776.816.39 frankincense7.436.866.49 Octagonal fennel7.596.796.72
    [169] As can be seen from the table, the pH change of the paper specimens due to the volatile extract of clove, frankincense, and octagonal fennel was not significantly different from the control.
    [170] All. Strength
    [171] In order to confirm the degree of damage of the paper by the fragrance of the volatile extracts, the results of the measurement of the fracture strength of two pretreated preservative papers, newspaper paper, and Korean paper are shown in the following table.
    [172] Preservation 2NewspaperAfter Control571.940.40235.7 cloves501.842.6799.4 frankincense601.9155.64558 Octagonal fennel491.931.47124.1
    [173] As can be seen from the table, there were no significant differences in the strength of cutout strength between two kinds of preservative papers and newspaper papers due to the volatile extract of clove, frankincense, and octagonal fennel. In the case of Hanji, there was a significant difference in the severity strength in all extracts compared to the control group.However, unlike Yangji, which is a mechanical paper, the paper was hand-floated by hand. It is the result that Hanji itself does not maintain uniform strength compared to sunny paper because it maintains the wet strength only by interaction of the fiber of buckwheat.
    [174] la. Ion Chromatography
    [175] 7 kinds of anion for pretreated preservation paper, newspaper paper, and Hanji paper to measure the chemical composition change of paper material by fragrance of volatile extract-, Cl-, NO2 -, Br-, NO3 -2, PO4 -3, SO4 -2of The result of having measured the concentration is shown in the following table.
    [176] samplePreservation 2 F - Cl - NO 2 - Br - NO 3 -2 PO 4 -3 SO 4 -2Control026.68000.68014.05 cloves028.62000.14014.34 frankincense029.74000.37014.95 Octagonal fennel027.18000.51013.86 sampleNewspaper F - Cl - NO 2 - Br - NO 3 -2 PO 4 -3 SO 4 -2Control0 000.780.0139.57 cloves0 000.640.0440.36 frankincense0 000.850.0438.53 Octagonal fennel0 00One0.0432.18 sampleAfter F - Cl - NO 2 - Br - NO 3 -2 PO 4 -3 SO 4 -2Control0 001.095.586.61 cloves0 00One2.557.51 frankincense0 001.062.919.19 Octagonal fennel0 001.602.827.23
    [177] As can be seen from the table, all the paper specimens did not show a significant difference from the control. However, the significant result shown in the nitrate anion (NO 3 -2 ) is due to the difference due to ammonium nitrate (NH 4 NO 3 ) added as a water active reagent.
    [178] ② Material stability result for fibers
    [179] end. Tensile strength
    [180] In order to measure the condition and the degree of damage of the fiber due to the fragrance component of the volatile extract, the results of the measurement of the actual tensile strength for silk, cotton, and embroidery yarns are shown in the following table.
    [181] sampleControlclovesfrankincenseOctagonal fennel Cotton yarn0.600.580.620.58 Silk thread0.890.890.860.87 Embroidery thread0.450.500.490.51
    [182] As can be seen from the above table, the actual tensile strength values of all the fibers pretreated with cloves, frankincense, and octagons were not significantly different from those of the control group.
    [183] As a result of investigating the effect of volatile extracts of herbal medicines on paper and fiber materials, volatile extracts of clove, frankincense, and octagonal fennel did not show significant difference in whiteness, acidity and stiffness compared to the control group. As a result of measuring the concentration of the seven anions using the component did not show a difference in concentration. In addition, there was no significant difference in the actual tensile strength of the three types of fiber yarns compared to the control group, and the fragrance components of the volatile extracts of clove, frankincense, and octagonal fennel did not damage paper and fiber materials. It was confirmed that the material was stable.
    [184] As described above, it can be seen that the volatile extracts and their mixtures have antibacterial and insecticidal effects on the main fungi and worms inhabiting cultural properties, and the synergistic effect of the volatile extracts compared to the volatile extracts. It can be seen that there is. Therefore, volatile extracts can be used alone or in combination according to the kinds of fungi and worms inhabiting the cultural property.
    [185] In addition, the biocide composition for the preservation of cultural properties as a volatile extract of one or more natural products selected from the group consisting of cloves, frankincense and octagonal fennel according to the present invention has been a cultural property of the cultural properties due to the influx of fungi and insects into the exhibition space In addition to physical control methods such as the installation of filters to block damage, it also enables effective preventive management against the damage of fungi and insects of organic cultural properties.
    [186] In addition, the biocide composition for preserving the cultural property of the present invention is effective in the sustained and stable cultural property is solved by solving the problem of persistence and toxicity of the drug, which is a problem of conventional control for preserving the cultural property.
    权利要求:
    Claims (12)
    [1" claim-type="Currently amended] A biocide composition for preserving cultural properties, comprising a volatile extract of at least one natural product selected from the group consisting of cloves, frankincense and octagonal fennel.
    [2" claim-type="Currently amended] The biocide composition according to claim 1, which comprises at least volatile extract of octagonal fennel.
    [3" claim-type="Currently amended] The biocide composition according to claim 1 or 2, wherein the octagonal volatile extract is 50% or more based on the total volume of the composition.
    [4" claim-type="Currently amended] The biocide composition according to claim 1 or 2, wherein the volatile extract is extracted by a solvent-distilled extraction method.
    [5" claim-type="Currently amended] The method of claim 1, wherein the cultural habitat organism is a fungus Trichoderma koningii , Mucor hiemalis , Aspergillus niger , Penicillium funniol Locust ( Penicillium funiulosum ), the larvae are termites ( Reticulitermes speratus ), thigh bark ( Lyctus linearis GOEZE) and rice weevil ( Sitophilus oryzae L.), characterized in that the biocide composition for cultural property preservation.
    [6" claim-type="Currently amended] The biocide composition for cultural property preservation according to claim 1, wherein the cultural property is an organic cultural property.
    [7" claim-type="Currently amended] The biocide composition according to claim 6, wherein the organic cultural property is a tributary or a fiber.
    [8" claim-type="Currently amended] The biocide composition according to claim 1, wherein the volatile extract of one or more natural products selected from the group consisting of cloves, frankincense and octagonal fennel contains eugenol or anetol.
    [9" claim-type="Currently amended] A method of preserving cultural property, characterized in that the cultural property is treated with the biocidal composition for cultural property preservation according to claim 1 or 8 by a direct spray method, a coating method, a fumigation method, an aroma treatment method or a packaging method.
    [10" claim-type="Currently amended] 10. The method of claim 9, wherein the cultural property is treated by aroma treatment.
    [11" claim-type="Currently amended] The method according to claim 10, wherein the effective amount of the aroma treatment method is 50 ml / m 3 or more in terms of concentration in air.
    [12" claim-type="Currently amended] 10. A method according to claim 9 having a formulation of an aerosol, tincture, liquid or matrix gel.
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    同族专利:
    公开号 | 公开日
    KR100421538B1|2004-03-18|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    法律状态:
    2001-01-29|Application filed by 대한민국 (관리부서:국립문화재연구소)
    2001-01-29|Priority to KR10-2001-0003996A
    2002-08-03|Publication of KR20020063375A
    2004-03-18|Application granted
    2004-03-18|Publication of KR100421538B1
    优先权:
    申请号 | 申请日 | 专利标题
    KR10-2001-0003996A|KR100421538B1|2001-01-29|2001-01-29|Biocide composition comprising volatile extracts of natural medicines as effective components for conservation of cultural properties, and method for conserving cultural properties using the same|
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