KR101819108B1 - Composition for Biofilm removal comprising Eugenol and Emulsifier as active ingradient for Stone Conservation and Method thereof for against Biological weathering - Google Patents

Abstract

The present invention relates to a composition for removing biofilm containing eugenol and an emulsifier as active ingredients. The composition for removing biofilm containing eugenol and the emulsifier as the active ingredients maintains stable dispersed state, shows antibacterial functions, and inhibits microbes and lichens formed on stone works outdoors. Thus, the composition for removing biofilm containing eugenol as the active ingredient can be useful for cleaning and preserving stone works from weathering.

Description

[0001] The present invention relates to a composition for removing biofilm containing eugenol and an emulsifier as an effective ingredient, and a method for preserving a marble from the biological weathering using the same.

The present invention relates to a composition for removing biofilm containing eugenol and an emulsifier as an active ingredient, and a method for removing biofilm on the surface of a masonry.

Stone has been widely used because it has good durability compared to other materials such as wood and metal, and is easily available. These kinds of stone artifacts are reminiscent of stone artifacts made in various places from prehistoric Bandal dolls to the stone pagoda erected in the Three Kingdoms period and the statues built in the tombs of the Joseon Dynasty. According to the statistics of the Cultural Heritage Administration by the end of December, 2016, the cultural assets made of stone in Korea are 80 national treasures, 521 treasures, 27 historic monuments, 14 important folk cultural assets, 32 registered cultural properties, 674 In total, about 15.2% of the total number (4,423 cases) is occupied, and 569 of the 601 of national treasures and treasures are classified as architectural cultural assets. However, due to the nature of the stone artifacts such as the tower, the buildings and the Buddha statues, many of them are exposed to the open air, so they are greatly affected by external environment such as temperature, humidity, rain and wind, air pollutants and living creatures. .

State-designated cultural property by material (Unit: case) National treasure treasure Historical Sightseeing natural
monument important
Folk
Cultural Heritage Registered cultural assets Sum Stone 80 521 27 - - 14 32 674 woodcarving 32 263 40 - - 184 27 546 metal 75 184 - - - - 53 312 Tozai 53 129 - - - - One 183 paper 61 676 - - - - 89 826 Other 27 287 428 109 456 96 479 1,882 Sum 328 2,060 495 109 456 264 681 4,423

The general weathering rate of stone is known to be very slow, but once the weathering progresses, peeling or peeling will occur very vigorously. It is known to be fundamentally affected by water, leaving weathered residues in response to water-rock reactions, and expanding its volume as the residues crystallize, accelerating physical or chemical weathering. Biological weathering occurs when organisms grow in stone cultural properties. It is caused by microbial growth from air and surrounding dust and soil, physical weathering caused by the root pressure applied by the roots of the surrounding vegetation, Weathering is known to occur at the same time. Photosynthetic autotrophs (photolithoautotrophs) that synthesize nutrients directly through photosynthesis such as algae, cyanobacteria, and lichens are grown on stone artifacts exposed to nature. When these organisms are grown and accumulated on the surface of rocks, heterotrophs that grow in energy by ingesting organic matter such as common bacteria or fungi grow, and when nitrogen compounds or sulfur compounds are present in the habitat, Chemolithoautotrophs that produce energy by using inorganic substances such as sulfuric acid bacteria are also generated. As these communities grow, hyphae of fungi and lichens physically penetrate into the rocks and gaps are formed, and the organic acids and metabolites generated during the metabolism of these organisms dissolve the minerals in the rocks, which can lead to destruction of the stone . In addition, biofilm formation by fungi, algae, and the like promotes serious surface contamination and secondary weathering of stone as well as damage by higher plants. As mentioned above, in the case of stone artifacts in the outdoors, it comes into contact with organisms in addition to the soil organic matter in the foundation, so that it is easily exposed to such biological weathering.

Because of the durability and ease of supply and demand of materials as described above, stone is widely used not only in domestic but also overseas, so that protection of stone cultural property from biological weathering is also required. Cambodia's Angkor Wat ruins The major causes of weathering in the stone cultural properties were chemical weathering of whiteness caused by the reaction of carbonate minerals with water, and biological weathering due to growth of lichens, birds and woody plants. In Spain, Carrascosa The del Campo cathedral suffered from the problem of biological weathering, such as the degradation and corrosion of the algae and heterotrophic bacteria and fungi, which accelerated the transition of organisms to higher organisms such as cetaceans and lower plants. A comparatively large number of studies have been carried out for the weathering and preservation of stone cultural properties in Korea, such as the identification of biofilm formation process and the classification of stone surface species in stone cultural properties composed of granite and sandstone. In the case of Korea, there is a hot and humid summer season in which the creatures can reproduce. Due to the change of four seasons, weathering by various organisms can easily occur. In 1990, the National Institute of Cultural Properties surveyed the status of lichen, And a chemical control process is applied to the process.

In order to preserve stone artifacts well, it is necessary to identify the cause of the damage one by one and to take measures to eliminate the cause of the stone artifacts, so that it is necessary to prevent further damage. In the preservation of stone artifacts, washing is one of the important processes performed basically. Washing is also an important step in advance for preservation treatment to prevent weathering progress such as reinforcement and waterproof treatment. In this case, the washing process should be performed in such a way that the stoneware itself is not damaged and the secondary damage by washing does not occur.

Various methods have been studied for methods and problems related to the cleaning of stoneware, and physical and chemical cleaning methods have been used. The mechanical method is a polishing method using abrasive materials such as silicon dioxide, non-silicon dioxide-based fine sand, and dry ice, which causes a change in the detail or texture of the stone due to the abrasive material itself, Chemical treatment with HF or sodium hydroxide may also result in immediate or long-term potential damage by chemicals contained in the solution. For example, immediate damage such as surface damage caused by detergents such as sulfuric acid may occur. In the long term, where algae and lichen are more active and reproducible by residues after a certain period of time due to phosphate (H ₃ PO ₄ ) Has been reported to have occurred.

Korean Patent Registration No. 10-0922608 discloses a method of preventing fungal weathering by mixing ethylene oxide and methyl bromide to prevent fungal weathering, There is a complexity and safety problem of the procedure such as the hypothesis of the scaffold for the scaffold and construction of the separate fumigation device and the step of exhausting the fumigation gas to the safe place after the treatment. In the case of using worn materials or chemicals, not only damage of stone, but also ease of work and safety of workers should be considered.

In this way, many researches on the factors causing physical weathering of stone cultural properties and physical and chemical removal methods have been conducted in Korea and abroad. However, researches and reports on environmentally friendly and low - risk removal techniques have been lacking.

Therefore, the inventors of the present invention based on an active ingredient of an environmentally friendly natural medicinal substance rather than a chemical treatment used in the conventional biofilm removal, and tried to develop a proper formulation to prevent the damage of the stone cultural property due to the physical washing method. In order to overcome the problem of biological weathering in stone cultural properties, previous studies have been carried out to investigate the natural medicinal herb extracts inhibiting the growth of biofilm formation bacteria, and the eugenol extracted from the clove has antimicrobial activity against biofilm forming microorganisms .

In addition, the present inventors have conducted a study of mixing various emulsifiers to increase the water solubility of yugenols, developed a composition for removing biofilm containing yugenol and an emulsifier which are dissolved in vigor, and stably exist as active ingredients, And the effect on the mudstone, thereby completing the present invention.

Korean Patent No. 10-0922608

It is an object of the present invention to provide a composition for removing biofilm containing eugenol and an emulsifier as an active ingredient.

In order to achieve the above object, the present invention provides a composition for removing biofilm containing eugenol and an emulsifier as an active ingredient.

The present invention also provides a method for removing biofilm on a surface of a stone including the step of treating the composition with a stoneware.

In addition, the present invention provides a method of cleaning a surface of a masonry comprising the step of treating the composition with a stone cultural property.

The composition for removing biofilms containing the eugenol and the emulsifier of the present invention as an active ingredient can be used as a biofilm for removing algae and microorganisms that are actually present in outdoor stones without causing damage to the stone while maintaining a stable dispersion state, Inhibitory effect. The composition for removing biofilm comprising the above eugenol as an active ingredient can be usefully used for preserving and washing stoneware from biological weathering.

1 to 8 show the results of the dispersing ability of yugunol according to the concentration of each emulsifier. (Fig. 1: Tween 20, Fig. 2: Tween 40, Fig. 3: Tween 80, Fig. 4: Span 20, Fig. 5: Span 80, Fig. 6: Propylene glycol,
9 to 11 are the results showing the Yugenol dispersing ability according to the emulsifier mixture. (Figure 9: Tween 20 + Tween 80, Figure 10: Span 20 + Span 80, Figure 11: Span 20 + Tween 20)
12 and 13 are the results of antimicrobial effect of the composition for removing biofilm according to the present invention on fungi collected from domestic stoneware.
14 and 15 are the results showing the antimicrobial effect of the composition for removing biofilm of the present invention on fungi collected from foreign stoneware.
Fig. 16 shows the results of the changes of the stone samples by the biofilm removing composition of the present invention.
A: Before the composition for removing the biofilm of the stone sample
B: After the composition for removing the biofilm of the stone sample was treated
C: Weight comparison before and after treatment of composition for removing biofilm of stone samples
17 is a mineralogical analysis result of a sample of a stone material by the composition for removing biofilm of the present invention and a control group of the present invention.
FIG. 18 is a graph showing the change of the outdoor stonework in Korea by the biofilm removal composition of the present invention.
A: composition for biofilm removal
B: Comparison of microscopic observation before / after treatment with composition for biofilm removal
C: Comparison of microbial activity before and after treatment with composition for removing biofilm
FIG. 19 shows the results of the change of the outdoor stoneware of the present invention with the biofilm removal composition of the present invention.
A: Comparison of overall appearance before and after treatment with composition for removing biofilm and control group
B: Detailed appearance change by composition for biofilm removal and control treatment
C: Comparison of microbial activity before and after treatment by composition for biofilm removal and control treatment

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for removing biofilm containing eugenol and an emulsifier as an active ingredient.

The eugenol is a compound represented by the following formula (1).

[Chemical Formula 1]

The eugenol is called clove ( Syzygium aromaticum ), wormwood (wormwood), cinnamon, sweet basil ( Ocimum but are not limited to, those derived from other materials, such as, for example, basilicum , star anise, lemon balm, vanilla, celery or ginger, Anything can be used.

The above eugenol may be used in the form of a biologically acceptable salt, and as the salt, an acceptable alkali addition salt is useful. The expression " biologically acceptable salt " means any organic or inorganic addition salt that has a relatively non-toxic, harmless effective action on the subject, and the side effects of this salt do not impair the beneficial effects of the eugenol. These salts include alkali metal salts (such as sodium salts and potassium salts) and alkaline earth metal salts (such as calcium salts and magnesium salts). For example, aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, oleamine, potassium, sodium, tromethamine, .

And the eugenol may be dispersed using the emulsifier. The emulsifier is a substance that is added to facilitate the preparation of the emulsion and to stably maintain the resulting emulsion, and usually a surfactant having at least one hydrophilic group and at least one hydrophobic group in one molecule acts as an emulsifier. In addition, a surfactant having high hydrophilicity is used as an emulsifier of O / W type emulsion, and a surfactant having high hydrophobicity is used as an emulsifier of W / O type emulsion.

The emulsifier may be selected from and mixed with two or more emulsifiers selected from the group consisting of Tween 20, Tween 40, Tween 80, Span 20, Span 80, propylene glycol, glycerin and lecithin, But is not limited thereto. Other examples of emulsifying agents include carbocyclic acids and salts thereof (such as alkaline soaps of sodium, potassium and ammonium, metallic soaps of calcium or magnesium, lauric, palmitic, stearic and oleic acids, Organic basic soaps), alkylphosphates or phosphoric esters, acid phosphates, diethanolamine phosphates, potassium cetylphosphates, ethoxylated carboxylic acids or polyethylene glycol esters, PEG-n acrylates, linear fatty alcohols having 8 to 22 carbon atoms (Branched from 2 to 30 moles of ethylene oxide and / or from 0 to 5 moles of propylene oxide with a fatty acid having 12 to 22 carbon atoms and an alkylphenol having 8 to 15 carbon atoms in the alkyl group) Fatty alcohol polyglycol ethers (e.g., Laureth-n, Cetearas-n, Steareth-n, Oleth-n) Glycol ethers (e.g., PEG-n stearate, PEG-n oleate, PEG-n cocoate); Monoglycerides and polyol esters; C12-C22 fatty acid mono-and di-esters of adducts from 1 to 30 moles of ethylene oxide with a polyol; Fatty acids and polyglycerols such as monostearate glycerol, diisostearoyl polyglyceryl-3-diisostearate, polyglyceryl-3-diisostearate, triglyceryl diisostearate, polyglyceryl- 2-sesquiisostearate or polyglyceryl dimerate (a mixture of compounds from a number of these substance classes is also suitable); Fatty acids and saccharose esters (such as sucrose esters), glycerol and saccharose esters (such as sucrose glycerides), sorbitol and sorbitol (such as sucrose esters), polyoxyethylene Sorbitan mono- and di-esters of saturated and unsaturated fatty acids having from 6 to 22 carbon atoms, and ethylene oxide adducts, polysorbate-n series, sorbitan esters (e.g., sesquiisostearate, Sorbitan, PEG- (6) -isostearate sorbitan, PEG- (10) -sorbitan laurate, PEG-17-diolate sorbitan), glucose derivatives, C8-C22 alkyl-mono and oligo- Seed, and an ethoxylated analogue of glucose, a O / W emulsifier (for example, methylglucose-20 sesquistearate, sorbitan (E.g., methyl glucoside diolate / methyl glucose isostearate), sorbitan monostearate (sorbitol), sorbitan monostearate (sorbitol) But are not limited to, sodium alginate, sodium alginate, sodium alginate, sodium alginate, sodium alginate, sodium alginate, sodium alginate, sodium alginate, sodium alginate, The present invention also relates to a method for producing a compound represented by the following formula (1): wherein R 1 is selected from the group consisting of an alkyl group, an alkenyl group, an alkenyl group, an alkenyl group, an alkynyl group, , Alkylamidazolines, oxadiamines having a chain containing a heterocycle, pyridine derivatives, isoquinotenes, cetylpyridinium chlora Such as cetyltrimethylammonium bromide (CTBA), stearylalkonium, amide derivatives, alkanolamides such as acylamide DEA, ethoxylated amides such as PEG-n acylamide, oxy Deamide; Polysiloxane / polyalkyl / polyether copolymers and derivatives, dimethicone, copolyols, silicone polyethylene oxide copolymers, silicone glycol copolymers, propoxylated or POE-n ethers (Meroxapol), poloxamers or poly (Oxyethylene) m-block-poly (oxypropylene) n-block (oxyethylene), a zwitterionic surfactant having at least one quaternary ammonium group and at least one carboxylate and / or sulfonate group in the molecule The zwitterionic surfactant is betaine, for example, N-alkyl-N, N-dimethyl ammonium glycinate, coco alkyldimethyl ammonium glycinate, N-acylaminopropyl-N, N-dimethyl ammonium glycinate, 2-alkyl-3-carboxymethyl-3-hydroxyethylimidazoline having 8 to 18 carbon atoms in each alkyl or acyl group, Also included within the scope of the invention are cocoamylaminoethyl hydroxyethyl carboxymethyl glycinate, N-alkyl betaine, N-alkyl amino betaine, alkyl imidazoline, alkyllopeptides, lipoamino acids, self emulsifying bases, and KFDePolo, A short textbook of cosmetology, Chapter 8, Table 8-7, p250-251. Nonionic emulsifiers such as PEG-6 beeswax (and) PEG-6 stearate (and) polyglyceryl-2-isostearate [Apifac], glyceryl stearate and and PEG-100 stearate [Arlacel 165] PEG-5 glyceryl stearate [arlatone 983 S], sorbitan oleate and polyglyceryl-3 ricinoleate [Arlacel 1689], sorbitan stearate and sucrose cocoate [arlatone 2121], glyceryl stearate Cetomacrogol Wax], cetearyl alcohol and colisorbate 60 and PEG-150 and Stearate-20 [Polawax GP 200, Polawax NF [Emulgade PL 1618], cetearyl alcohol and Cetearyl-20 [Emulgade 1000NI, Cosmowax], cetearyl alcohol and PEG-40 castoryl alcohol [Emulgade F Special] , Cetearyl alcohol and PEG-40 castor oil and sodium cetearate [Emulgade F], stearyl alcohol and stearyl-7 and stearase-10 [Emulgator E 2155], cetearyl alcohol and stearyl-7 and stearase-10 [Emulsifying wax USNF], glyceryl stearate Propyleneglycol iseseth-3 acetate [Hetester PHA], cetearyl alcohol and Ceteth-12 and oles-12 and PEG-75 stearate Gelot 64, propylene glycol ceteth-3 acetate [Hetester PCS] [Lanfitol Wax N 21], PEG-6 stearate and PEG-32 stearate [Tefose 1500], PEG-6 stearate and Ceteth-20 and Stearase-20 [Tefose 2000] TES-20 and glyceryl stearate and Stearase-20 [Tefose 2561], glyceryl stearate and Cetearase-20 [Teginacid H, C, X].

The emulsifying agent used in the composition of the present invention may be Tween 20 and Span 20.

The emulsifier Tween 20 and Span 20 may be mixed at a ratio of 6 to 8: 2 to 4.

The emulsifier may have a hydrophilic-lipophilic balance (HLB) value of 13 to 15

The composition for removing biofilm may comprise 4 to 6 parts by weight of eugenol, 8 to 12 parts by weight of an emulsifier, and 80 to 90 parts by weight of distilled water.

The composition for removing the biofilm containing eugenol and an emulsifier as an active ingredient may be prepared in any of the formulations conventionally produced in the art, and may be, for example, a foam, a gel, a soap or a powder , Preferably in a liquid state.

The biofilm removal may be to remove the lichens and microorganisms on the surface of the stone.

In the concrete examples of the present invention, various emulsifiers were stably dispersed in various concentrations at different concentrations (Figs. 1 to 11), and the composition for removing biofilm having the above composition showed an inhibitory effect in experiments using fungi 12 to 15). The biofilm removing composition was found not to cause damage to the stone (Figs. 16 to 17), and showed a sustained inhibitory effect on lichens and microorganisms when used in the actual outdoor stoneware (Figs. 18 to 19).

Therefore, the composition containing eugenol and an emulsifier according to the present invention as an effective ingredient can be usefully used as a composition for removing biofilm.

The present invention also provides a method for removing biofilm on a surface of a stone surface, comprising the step of treating a stone cultural property with a composition containing eugenol and an emulsifier as active ingredients.

The organism may be a licentiate or a microorganism, and may be any microorganism that forms a biofilm on the surface of a stone to cause surface contamination or secondary weathering.

The composition can be used pure or diluted to a concentration suitable for use.

The composition can be applied to the stone cultural property by a direct spraying method, a coating method, a fumigation method, an orienting method, or a packing method, but is not limited thereto. The time for application to the stone cultural property is preferably, but not limited to, a clear day in the afternoon on which rain does not occur.

Formulations for applying the composition to the stone cultural property include, but are not limited to, aerosols for direct spraying, tinctures for fumigation treatment, liquid solutions, and matrix gels for sustained release formulations.

The application effective amount of the composition varies depending on the components and the application means. For example, in the aromatic treatment method, the concentration in the air is usually adjusted to 50 ml / m ³ or more, preferably 125 ml / m ³ or more.

The present invention also provides a method for cleaning a surface of a stone surface comprising treating a stone cultural property with a composition for removing biofilm containing eugenol and an emulsifier as an effective ingredient.

In the specific examples of the present invention, the composition for stable dispersion of yogurol was determined by varying the concentration of various emulsifiers (Figs. 1 to 11), and the composition for removing biofilm having the above composition showed an inhibitory effect in experiments using fungi 12 to 15). The biofilm removing composition was found not to cause damage to the stone (Figs. 16 to 17), and it was found that the biofilm-removing composition has a sustained inhibitory effect on algae and microorganisms by spraying on the actual outdoor stones (Figs. 18 to 19).

Therefore, the method of treating the biofilm-removing composition containing the eugenol and the emulsifier according to the present invention as an active ingredient to the mudstone can be effectively used for removing and cleaning the biofilm on the surface of the mudstone.

Hereinafter, the present invention will be described in detail with reference to examples.

EXAMPLES The following Examples and Experiments are for the purpose of illustrating the present invention, but the present invention is not limited by the following Examples and Experimental Examples.

Example  1. Emulsifier selection and mixing ratio search

Eight kinds of emulsifiers known to be edible or eco - friendly were selected for the selection of emulsifiers to confirm the emulsifying and dispersing power for yugenols. The emulsifiers used were Tween 20, Tween 40, Tween 80, Span 20, Sorbitan Monooleate, Propylene Glycol, 99% polyoxyethylene sorbitan monolaurate, ), Glycerin (99%), and lecithin. Sigma-Aldrich used a 99% pure standard reagent.

The emulsifier was added 3%, 5%, 7%, and 10%, respectively, and the remaining volume was filled with distilled water and mixed to observe the milky emulsification phenomenon. In order to accurately determine the degree of emulsification, the dispersions of yugenols and emulsifiers were observed using a congo red dye.

As a result, as shown in FIGS. 1 to 3, the Tween series having a high hydrophilic-lipophilic balance (hydrophilic-lipophilic balance) showed a phenomenon in which the eugenol was dispersed in water. However, As shown in FIGS. 4 and 5, when the HLB was low in the Span series, the lipophilicity was high and the mixture was well mixed with yugenol but was not mixed with water. Likewise, in the case of propylene glycol (FIG. 6), glycerin (FIG. 7) and lecithin (FIG. 8) having low HLB, they are mixed with yugenol but are not mixed with water and emulsified. Appeared quickly.

Example  2. Determination of mixing ratio of emulsifier

Based on the results obtained in Example 1, various HLB conditions were made as shown in Table 2 using a Tween series emulsifier to find an appropriate HLB.

Mixing concentration ratio (by volume ratio) Tween 20
(HLB 16.7) One 3 5 7 9 Tween 80
(HLB 15.0) 9 7 5 3 One HLB 15.17 15.51 15.85 16.19 16.53

 As a result of the mixing, separation was observed over time as shown in FIG. 9, and similar results were obtained in a mixture of propylene glycol and glycerin having a low HLB of about 4. Thus, it was confirmed that HLB was inadequate, and using Span series emulsifier, HLB conditions were created.

Mixing concentration ratio (by volume ratio) Span 20
(HLB 8.6) One 3 5 7 9 Span 80
(HLB 4.3) 9 7 5 3 One HLB 4.73 5.59 6.45 7.24 8.17

As a result, as shown in FIG. 10, the separation phenomenon appeared even when the HLB was 8 or more. To adjust the HLB in the range of 8 to 15, Span 20 and Tween 20 were mixed as shown in Table 4.

Mixing concentration ratio (by volume ratio) Span 20
(HLB 8.6) One 3 5 7 9 Tween 20
(HLB 16.7) 9 7 5 3 One HLB 15.89 14.27 12.65 11.03 9.41

As a result, as shown in FIG. 11, when Span 20 and Tween 20 were mixed at a ratio of 3: 7 and the HLB was adjusted to 14.27, precipitation and separation did not appear even after 1 day of mixing. Thus, Span20: Tween20 = 3: The mixing ratio of the cleaning composition was found to be Yugenol: emulsifier: dispersion medium = 5:10:85.

Example  3. A composition for removing biofilm containing eugenol and an emulsifier as an active ingredient Antimicrobial  Verify Active

Fungi in microorganisms are known to be one of the major causes of biological weathering that causes mycelial damage and physical damage in addition to damages caused by their own metabolites. The effect of the composition for removing biofilm including the mixture of eugenol and emulsifier obtained through Example 2 was confirmed.

Example  3-1. For fungi collected from domestic stoneware Antimicrobial  Verify Active

In order to confirm the effect of the composition for removing biofilm, the effect of the biofilm removing composition containing the eugenol and emulsifier of the present invention as an active ingredient was confirmed on seven fungi collected from the mudstone of Youngryung in Yeoju city, Kyonggi Province.

Specifically, Tween 20 and Span 20 were mixed in a ratio of 7: 3 as in Example 2 to prepare an emulsifier. The emulsifier, eugenol, and sterilized distilled water were mixed at a ratio of 10: 5: 85, To prepare a composition for removing biofilm. The samples collected from Youngryung were suspended in sterilized distilled water and cultured for 5 days at 28 ° C in a PDA medium (Potato Dextrose Agar: 20.0g of Potato dextrose broth, 18.0g of Agar, 50ppm of Chloram phenicol, 20ppm of Nalidixic acid and 1000ml of distilled water) After colonies were identified by conventional methods in the industry, seven representative fungi were selected as shown in Table 5. Each of the selected fungi was suspended in a suspension of 100 μl each of the colonies to prepare a suspension, which was then suspended in Czapek`s agar medium ( ₃ g NaNO ₃ , 1 g K ₂ HPO ₄ , 0.5 g MgSO ₄ .7H ₂ O, 0.5 g KCl, FeSO ₄ .7H ₂ O, Sucrose 30g, Chloram phenicol 50ppm, Nalidixic acid 20ppm, Agar 18g, and distilled water 1000ml), and a sterilized 8mm paper disk was placed on a culture plate. The composition for removing the biofilm was dropped on a paper disk at 50 후 and incubated at 28 캜 for 5 days. The inhibition zone was then measured. As a control group, distilled water and emulsifier (distilled water: emulsifier = 90 : 10) were respectively scored and compared in the same way.

Comparison of Growth Inhibition Cycle of Fungi Collected from Domestic Stoneware (Unit: mm)
inhibition zone (mm) Activity Distilled water Emulsifier Emulsifier + Yugenol Distilled water Emulsifier Emulsifier + Yugenol Trichoderma harzianum 0 0 8.33 - - ++ Trichoderma atroviride 0 0 15.67 - - +++ Purpureocillium lilacinum 0 0 15.00 - - +++ Pestalotiopsis sp . 0 0 17.67 - - +++ Penicillium oxalicum 0 0 15.00 - - +++ Diaporthe melonis 0 0 20.00 - - +++ Aspergillus aculeatus 0 0 8.00 - - ++

As a result, as shown in Table 5 and FIG. 12 to FIG. 13, no growth-inhibitory ring formation was observed in the control group of distilled water and emulsifier-treated group, but a strong antifungal activity was confirmed in the detergent mixed with emulsifier and yugenol

Example  3-2. Overseas  For fungi collected from stoneware Antimicrobial  Verify Active

In order to confirm that the effect of the composition for removing biofilm can be applied to other fungi living in foreign stoneware, 12 kinds of fungi were selected in the same manner as in Experimental Example 1-1 with respect to the samples collected from the ruins of Wat phou in Laos And the antifungal activity of the composition for removing biofilm was compared.

Comparison of growth inhibition of fungi collected from foreign stoneware (Unit: mm) Inhibition zone (mm) Activity Distilled water Emulsifier Emulsifier +
Yugenol Distilled water Emulsifier Emulsifier +
Yugenol Trichoderma harzianum 0 0 7.67 - - ++ Phoma sp . 0 0 9.00 - - ++ Penicillium simplicissimum 0 0 8.33 - - ++ Penicillium citrinum (LF8) 0 0 7.00 - - ++ Penicillium citrinum (LF1) 0 0 6.67 - - ++ Neosartoria fischeri 0 0 15.00 - - +++ Lasiodiplodia theobromae 0 0 0.67 - - + Fusarium sp . 0 0 6.33 - - + Fusarium solani 0 0 6.33 - - + Fusarium oxysporum 0 0 7.33 - - ++ Aspergillus sp . 0 0 7.33 - - ++ Aspergillus fumigatus 0 0 8.00 - - ++

As a result, as shown in Table 6 and FIGS. 14 to 15, no growth-inhibitory ring formation was observed in the control group of distilled water and emulsifier-treated group, but a strong antifungal activity against most of the fungi was observed in the detergent mixed with emulsifier and yugenol

Example  4. Stability evaluation for stone

The composition for removing biofilm with antimicrobial activity confirmed in Example 3 was treated with a composition for removing biofilm for evaluating the material stability for the effect on the stone when used in actual stone, .

Specifically, 10 ml of the composition for removing biofilm prepared in Example 2 and granite sample (3 to 4 g) were granulated at 25 ° C for 15.6 days, Considering the removal of chemicals due to rainfall at the time of treatment, the average value of rainfall days in the last 10 years was calculated and reflected. After the treatment, the stone sample and the composition for removing biofilm were centrifuged at 5,000 rpm for 10 minutes to recover a stone sample. The stone sample was washed twice with distilled water, centrifuged at 5,000 rpm for 20 minutes to remove the remaining biofilm removing composition, Lt; 0 > C for 24 hours. The surface state and color of the dried sample were confirmed with a stereomicroscope (Nikon SMZ1500). In addition, the weight of the emulsion was measured to 0.0001 g with a precision electronic balance (Shimadzu AW320), and the weight change before and after the treatment was compared using the emulsified group treated with distilled water and the untreated group as the control group.

As a result, it was confirmed that there was no difference between before and after the chemical reaction as shown in Fig. 16, and it was confirmed that the weight change was also within 0.15% and there was no apparent change.

X-ray diffractometer (Rigaku D / max-2200) was applied to the stone samples obtained by the above method in order to observe the changes in the composition of the samples in terms of mineralogy. The currents were 40kV and 40mA, respectively. The 2θ was set to 5˚ ~ 70˚ at 2˚ / min, and the control group was set up as in the external control group.

As a result, as shown in FIG. 17, there were no differences in feldspar, quartz, and biotite, which are major minerals forming granite, in both the treated and control groups, and it was confirmed that they do not cause damage or change in mineralogical aspects due to the tide of the present invention.

Experimental Example  1. Identification of the inhibitory effects of lichen and microorganisms adhered to actual outdoor stonework

Experimental Example  1-1. Confirmation of effect on domestic stone

The effect of the biofilm removing composition obtained through the above examples on actual outdoor stonework was confirmed.

Specifically, 50 ml of a detergent prepared according to the above Example 2 was sprayed on the whole stone of the stone material (35 cm × 23 cm × 95 cm) located in the Korean traditional culture university by spraying once. Precise photographing, stereomicroscopic photographing, and ATP measurement for surface contamination were carried out at the same position before, 3 weeks after, and 5 months after the treatment, and detailed measurements were carried out according to the manual of the manufacturer .

As a result, as shown in FIG. 18, at the same position, four places were observed at a magnification of 50 at a magnification of 50. As a result, white stippled pathogens were struck off and partly exposed to the lower part of the algae layer. It was observed that the algae discolored with yellowing. In addition, it was confirmed that cracks were enlarged as the paper body was dried as a whole, and the powder formed on the surface of the paper body was damaged and fell off from the surface, so that the paper body was exposed and some paper bodies were found to be damaged. It was also confirmed that the red stippling pathogens formed between the rocks were also lost.

In addition, the measurement results of the changes in the five locations in the same position before and after the treatment using the ATP measuring device (3M Clean-trace). At 3 weeks after treatment, the decrease in ATP was 88.8%, 98.0% in 2, 96.2% in 3, 43.9% in 5, and 95.5% in 5, respectively. Surface microbial activity was low.

Experimental Example  1-2. Overseas  Confirmation of effect on stone

Cambodia does not show much rain during November and March when it is cold and dry in the northeast monsoon season, but from April to May it starts from May to October with strong winds, moisture and rain, . In the warmest April, the maximum temperature rises to 38 ° C. In the coldest January, the maximum temperature rises to 20 ° C.

In order to confirm the effect of the composition for removing biofilm according to the present invention under these different climatic conditions, 50 ml of the composition for removing biofilm was sprayed to the whole stoneware with a sprayer on a stone member in Preah Pithu, Cambodia , 2 months (dry season), 5 months (dry season) and 8 months (wet season).

As a result, as shown in FIG. 19, the measurement area was dry and the color of the lichen was changed overall, but the color change was more prominent in the treated stone for biofilm removal. In the case of the biofilm removal composition, it was observed that the leaves of the foliar pathogens fell off from the surface of the rock while the emulsifier, distilled water, and untreated control group showed color change only, I did. Even after 8 months of wet weather, which is a favorable climate for bio-growth, the biofilm removal composition treatment group showed extremely low lichen area compared to other treatment groups.

In addition, ATP measurement was performed to confirm surface contamination, and compared with before treatment and 2 months after treatment at the same site. In addition to the effect on the lichen treatment, ATP measurement value was 60.1% And it was found that the composition for removing biofilm of the present invention has an inhibitory effect on microorganisms and lichens even under different climatic conditions in Korea.

Claims (10)

Eugenol and
Tween 20 and Span 20 are mixed in a ratio of 6 to 8: 2 to 4 as an active ingredient,
Wherein the emulsifier has a hydrophilic-lipophilic balance (HLB) value of 13 to 15.
delete delete delete The composition for removing biofilm according to claim 1, wherein the composition contains 4 to 6 parts by weight of eugenol, 8 to 12 parts by weight of an emulsifier, and 80 to 90 parts by weight of distilled water.
The composition for removing biofilm according to claim 1, wherein the biofilm removal process removes the lichen and microorganisms on the surface of the stone.
A method for removing biofilm on a surface of a stone surface comprising treating the composition of claim 1 to a stone cultural property.
The method of claim 7, wherein the organism is a lichen or a microorganism.
The method according to claim 7, wherein the composition is treated by a spraying method, a coating method, a fumigation method, an orientation treatment method or a packing method to a stone cultural property.
A method for cleaning a surface of a masonry comprising the step of treating the composition of claim 1 to a stone cultural property.

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