JP6775181B2 - Concomitant agent containing terpene aldehyde - Google Patents

Description

The present invention relates to an anti-microbially active composition containing terpenaldehyde.

Although the inside of shoes used in daily life is contaminated with bacteria and fungi, there is a problem that there are few appropriate sterilization methods. In particular, many people are concerned about the stench of stuffy shoes regardless of the season. In addition, since the tinea bacillus in the shoes of tinea pedis patients grows for a long period of time, it has been pointed out that even if the foot is treated, it will be re-infected by ping pong infection. In addition, it has been shown that it is effective to sterilize and sterilize viruses, bacteria, and fungi floating in the air in closed spaces such as schools and workplaces as measures against various infectious diseases. Although artificially synthesized anti-microbial active substances have been mainly used for this purpose, artificially synthesized substances have side effects other than anti-microbial activity, environmental pollution, generation of resistant bacteria, and the like. There is also a problem, and a preparation having naturally derived antimicrobial activity is required.

Therefore, an object of the present invention is to provide a preparation composed of a naturally occurring substance having excellent antimicrobial activity. In particular, an object of the present invention is to provide a composition comprising a combination of naturally occurring components that exert strong antimicrobial activity when used in combination.

As a result of diligent studies, the present inventors have found that by using a first aldehyde, which is an acyclic terpen aldehyde, and an aldehyde different from the first aldehyde in combination, an excellent antimicrobial effect is exhibited. .. In particular, we have found that the combination of citral with an aldehyde selected from acyclic aliphatic aldehydes, terpene aldehydes and aromatic aldehydes exhibits synergistic antimicrobial activity. In addition, the present inventors have found that these concomitant composition can exert an antimicrobial effect on a space or a surface that is not in direct contact with each other due to volatilization, and have completed the present invention.

Specifically, the present invention relates to the following invention:
(1) A first aldehyde which is an acyclic terpenaldehyde and a second aldehyde which is different from the terpenaldehyde and is selected from an acyclic aliphatic aldehyde, a terpenaldehyde and an aromatic aldehyde. An anti-microbial active composition contained as an active ingredient.
(2) The antimicrobial active composition according to (1), wherein the first aldehyde and the second aldehyde are components derived from plant essential oils.
(3) The anti-microbial active composition according to (1), wherein the first aldehyde is citral.
(4) The antimicrobially active composition according to any one of (1) to (3), wherein the second aldehyde is linear or branched C6-12 alkanal.
(5) The antimicrobial active composition according to (4), wherein the second aldehyde is a linear or branched C8-10 alkanal.
(6) The antimicrobially active composition according to (5), wherein the second aldehyde is octanal, nonanal, decanal, or trimethylhexanal.
(7) The antimicrobial active composition according to any one of (1) to (3), wherein the second aldehyde is a linear, branched or cyclic C8-12 terpene aldehyde.
(8) The antimicrobial active composition according to any one of (1) to (3), wherein the second aldehyde is a linear, branched or cyclic C10 terpene aldehyde.
(9) The antimicrobial active composition according to (8), wherein the second aldehyde is citronellal or perillaldehyde which may be substituted with a hydroxyl group.
(10) The second aldehyde is a benzaldehyde which may be substituted with one or two substituents, or a cinnamaldehyde which may be substituted with one or two substituents, and here, The antimicrobial active composition according to any one of (1) to (3), wherein the substituent is a group selected from the group consisting of a hydroxyl group, a C1-4 alkyl group, and a C1-4 alkoxy group. ..
(11) The second aldehyde is a benzaldehyde or a cinnamaldehyde which may be substituted with one or two substituents, wherein the substituents consist of a hydroxyl group, a propyl group and a methoxy group. The anti-microbial active composition according to (10), which is a group selected from the group.
(12) The antimicrobial activity composition according to any one of (1) to (11), which is a volatile preparation.
(13) The anti-microbial activity composition according to (12), which has anti-microbial activity against microorganisms in the air.
(14) The antimicrobial activity composition according to any one of (1) to (11), which is a liquid preparation for spraying.
(15) The antimicrobial active composition according to any one of (1) to (14), which is used for deodorizing.
(16) The antimicrobial activity composition according to any one of (1) to (15), wherein the microorganism is a virus, a bacterium or a fungus.
(17) The antimicrobial active composition according to (16), wherein the microorganism is Bacillus subtilis, Trichophyton, or Candida.

As used herein, the term "terpene aldehyde" is also referred to as a terpene-based aldehyde, and is an aldehyde having an isoprene structure, and includes acyclic terpene aldehyde and cyclic terpene aldehyde. In the present specification, the "acyclic terpene aldehyde" is a terpene aldehyde that does not contain a cyclic structure, and examples thereof include citral (geranial, neral), citronellal, and hydroxycitronellal. In the specification, the "cyclic terpene aldehyde" is a terpene aldehyde having a cyclic structure, and perillaldehyde can be mentioned.

As used herein, the term "acyclic aliphatic aldehyde" means a linear or branched alkyl group having no cyclic structure and substituted with a formyl group (CHO). Examples of the acyclic aliphatic aldehyde include C1-20 alkanal, C6-12 alkanal, C8-10 alkanal, and the like, and include metanal, ethanal, propanal, isopropanal, butanal, isobutanal, and tert. − Butanal, pentanal, 2-methylbutanal, 3-methylbutanal, 2,2-dimethylpropanal, hexanal, 2-methylpentanal, 3-methylpentanal, 4-methylpentanal, 2,2-dimethyl Butanal, 2,3-dimethylbutanal, 3,3-dimethylbutanal, 2-ethylbutanal, 3-ethylbutanal, heptanal, 2-methylhexanal, 3-methylhexanal, 4-methylhexanal, 5- Methylhexanal, 2,2-dimethylpentanal, 2,3-dimethylpentanal, 2,4-dimethylpentanal, 3,3-dimethylpentanal, 3,4-dimethylpentanal, 4,4-dimethylpentanal , 2-Ethylpentanal, 3-Ethylpentanal, 2-propylpentanal, 2,2,3-trimethylbutanal, 2,3,3-trimethylbutanal, 2-Methyl2-ethylbutanal, 2- Ethyl 3-methylbutanal, octanal, 2-methylheptanal, 3-methylheptanal, 4-methylheptanal, 5-methylheptanal, 6-methylheptanal, 2,2-dimethylhexanal, 2,3- Dimethylhexanal, 2,4-dimethylhexanal, 2,5-dimethylhexanal, 3,3-dimethylhexanal, 3,4-dimethylhexanal, 3,5-dimethylhexanal, 4,4-dimethylhexanal, 4,5-dimethyl Hexanal, 5,5-dimethylhexanal, 2-ethylhexanal, 3-ethylhexanal, 4-ethylhexanal, 2,2,3-trimethylpentanal, 2,2,4-trimethylpentanal, 2,3,3- Trimethylpentanal, 2,3,4-trimethylpentanal, 2,4,4-trimethylpentanal, 3,3,4-trimethylpentanal, 3,4,4-trimethylpentanal, 2-methyl2-ethyl Pentanol, 2-methyl3-ethylpentanal, 2-ethyl3-methylpentanal, 3-ethyl3-methylpentanal, 2-ethyl4-methylpentanal, 3-ethyl4-methylpentanal , 2-propylpentanal, 2,2,3,3-tetramethylbutanal, 2,2,3,4-tetramethylbutanal, 2,2,4,4-tetramethylbutanal, 2,3 3,4-Tetramethylbutanal, 2,3,4,5-tetramethylbutanal, 3,3,4,5-tetramethylbutanal, 2,2-diethylbutanal, nonanaal, 2-methyloctanal , 3-Methyloctanal, 4-Methyloctanal, 5-Methyloctanal, 6-Methyloctanal, 7-Methyloctanal, 2,2-dimethylheptanal, 2,3-dimethylheptanal, 2,4 -Dimethylheptanal, 2,5-dimethylheptanal, 2,6-dimethylheptanal, 3,3-dimethylheptanal, 3,4-dimethylheptanal, 3,5-dimethylheptanal, 3,6-dimethyl Heptanal, 4,4-dimethylheptanal, 4,5-dimethylheptanal, 4,6-dimethylheptanal, 5,5-dimethylheptanal, 5,6-dimethylheptanal, 6,6-dimethylheptanal , 2-Ethylheptanal, 3-Ethylheptanal, 4-Ethylheptanal, 5-Ethylheptanal, 2,2,3-trimethylhexanal, 2,2,4-trimethylhexanal, 2,2,5-trimethyl Hexanal, 2,3,3-trimethylhexanal, 2,3,4-trimethylhexanal, 2,3,5-trimethylhexanal, 2,4,4-trimethylhexanal, 2,4,5-trimethylhexanal, 2,5 , 5-trimethylhexanal, 3,3,4-trimethylhexanal, 3,3,5-trimethylhexanal, 3,4,4-trimethylhexanal, 3,4,5trimethylhexanal, 3,5,5-trimethylhexanal, 2-Methyl 2-ethylhexanal, 2-methyl3-ethylhexanal, 2-methyl4-ethylhexanal, 2-ethyl3-methylhexanal, 3-methyl3-ethylhexanal, 3-methyl4-ethylhexanal, 2- Ethyl 4-methylhexanal, 3-ethyl4-methylhexanal, 4-ethyl4-methylhexanal, 2-propylhexanal, 3-propylhexanal, 2,2,3,3-tetramethylpentanal, 2,2,3 , 4-Tetramethylpentanal, 2,2,3,5-Tetramethylpentanal, 2,2,4,5- Tetramethylpentanal, 2,2,4,5-tetramethylpentanal, 2,3,3,4-tetramethylpentanal, 2,3,3,5-tetramethylpentanal, 2,3,4 4-Tetramethylpentanal, 2,3,4,5 Tetramethylpentanal, 2,3,5,5-Tetramethylpentanal, 3,3,4,4-Tetramethylpentanal, 3,3,4 , 5-Tetramethylpentanal, 3,3,5,5-Tetramethylpentanal, 3,4,5,5-Tetramethylpentanal, 3,4,5,5-Tetramethylpentanal, 4,4 , 5,5-Tetramethylpentanal, 2,2-diethylpentanal, 2,3-diethylpentanal, 3,3-diethylpentanal, 2-methyl2-propylpentanal, 3-methyl2-propylpentan Includes nal, 4-methyl2-propylpentanal, decanal, undecanal, dodecanal, tridecaneal, tetradecaneal, pentadecanal, hexadecaneal, heptadecanal, octadecaneal, nonadecanal, and icosanal. The acyclic aliphatic aldehyde is preferably a naturally occurring aldehyde, more preferably octanal, nonanal, decanal, and trimethylhexanal.

As used herein, the term "aromatic aldehyde" means a substance in which an aromatic hydrocarbon ring is replaced with an aldehyde group (including a formyl group). Examples of the aromatic hydrocarbon ring include benzene, naphthalene, azulene, anthracene, chrysene, pyrene, coronen, and keclen, and benzene is preferable. Examples of the aldehyde group contained in the aromatic hydrocarbon ring as a substituent include a formyl group, a C1-6 alkanol group, a C2-6 alkenal group, and a C2-6 alkynal group, preferably a formyl group and a C1. It is a -4alkanal group, a C2-4alkenal group, and a C2-4alquinal group, more preferably a formyl group or a propenal group. The aromatic hydrocarbon ring may be substituted with a substituent other than one or two aldehyde groups, and such substituents include a hydroxyl group, a straight chain or a branched C1-6 alkyl group, and a straight chain. Alternatively, a branched C1-6 alkoxy group can be mentioned, preferably a hydroxyl group, an isopropyl group, or a methoxy group. The aromatic aldehyde is preferably a naturally occurring aldehyde. The aromatic aldehyde may be, for example, benzaldehyde which may be substituted with 1 or 2 substituents, or cinnamaldehyde which may be substituted with 1 or 2 substituents, and preferably. Benzaldehyde, cinnamaldehyde, cuminaldehyde, and vanillin.

In the present specification, the "anti-microbial active composition" is not particularly limited as long as it is a composition used for suppressing the growth of microorganisms, but is intended to be ingested into the human body. It is not a thing, preferably the surface of the living body's skin (for example, feet, hands, etc.), an object that comes into direct contact with the living body's skin such as shoes / socks and clothing, or a space such as indoors or in a car (particularly closed It is a composition used to suppress the growth of microorganisms in the space. Therefore, the antimicrobial activity composition of the present invention is not only a composition having antimicrobial activity against microorganisms existing on the surface to which the composition is applied, but also against microorganisms in the air in the space where the composition is present. Contains a composition with antimicrobial activity. Therefore, the anti-microbial active composition of the present invention can be a deodorant or an antibacterial agent.

As an example, the antimicrobial active composition of the present invention can contain the first aldehyde and the second aldehyde in an amount of 1 to 10 mg / ml (preferably 1 to 3 mg / ml), respectively. Alternatively, the anti-microbial active composition of the present invention contains the first aldehyde and the second aldehyde in an amount of 1 to 15% (w / v) (preferably 2 to 6% (w / v)), respectively. can do.

The dosage form of the antimicrobial active composition of the present invention is not particularly limited as long as it is in the form used for the purpose of the present invention, but may be, for example, a volatile preparation or a coating preparation. Specific examples include a liquid, a spray, a gel, a gel-forming kit, a polymer, or a form impregnated with a porous material.

As used herein, "antimicrobial activity" means suppressing the growth of microorganisms and does not necessarily require killing the microorganisms. "Microbial" means viruses, bacteria and fungi, including bacteria of the genus Basilus, Staphylococcus, Trichophyton, Microsporm, Epidermophyton and / or fungi of the genus Candida.

The first aldehyde which is the acyclic terpene aldehyde of the present invention and the second aldehyde which is different from the terpene aldehyde and is selected from the acyclic aliphatic aldehyde, the terpene aldehyde and the aromatic aldehyde are used. Since the anti-microbial activity composition contained as an active ingredient is excellent in anti-microbial activity as compared with the case of using it as a single agent, it can be used as an antibacterial agent or a deodorant. Further, the antimicrobial active composition containing a naturally derived aldehyde of the present invention can provide an antibacterial agent or a deodorant having less burden on the environment because the active substance is a naturally derived component. ..

(A) It is a figure and a photograph which showed the procedure of the antibacterial effect test by the combined use of citral and perilaldehyde against Bacillus subtilis (Bacterial bacillus). (B) It is a graph which plotted the result of the antibacterial effect against Bacillus subtilis (Bacterial bacillus) by the combined use of citral and perylaldehyde for each concentration which becomes score 0. The vertical axis shows the perylaldehyde concentration (mg / ml), and the horizontal axis shows the citral concentration (mg / ml). The asterisk indicates a point where FIC index = 0.5 (synergistic effect). (C) It is a photograph of a colony formed as a result of an antibacterial test against Bacillus subtilis (Bacterial bacillus) by a combination of citral and perylaldehyde. (A) It is a figure and a photograph which showed the procedure of the combined effect test of perillaldehyde and citral against Trichophyton (Arthroderma vanbreuseghemii TIMM2789). (B) It is a graph which plotted the result of the antibacterial effect against tinea bacillus (Arthroderma vanbreusehemii TIMM2789) by the combined use of perylaldehyde and citral for each concentration which becomes score 0. The vertical axis shows the perylaldehyde concentration (mg / ml), and the horizontal axis shows the citral concentration (mg / ml). The asterisk indicates that FIC index = 0.375 (synergistic effect). (A) It is a figure showing the concentration and position of the test substance dropped on the thick filter paper arranged on the surface of an agar medium in the combined effect test of perylaldehyde and citral on the growth of Bacillus subtilis (Bacterial bacillus). (B) It is a photograph of a chalet obtained as a result of a combined effect test of perylaldehyde and citral on the growth of Bacillus subtilis (Bacterial bacillus). The left shows the results of the control test (perylaldehyde alone), and the right shows the results of the combined test of perylaldehyde and citral. The upper figure shows the concentration and position of the test substance dropped in each petri dish. (A) It is a figure showing the concentration and position of the test substance dropped on the thick filter paper arranged on the surface of the agar medium in the combined effect test of various aldehydes and citral on the growth of Candida albicans (TIMM1768). (B) It is a photograph of a petri dish obtained as a result of a combined effect test of various aldehydes and citral on the development of Candida albicans (TIMM1768). The left shows the results of the control test (vanillin alone), and the right shows the results of the combination test of vanillin and citral. The upper figure shows the concentration and position of the test substance dropped in each chalet. (A) It is a figure which shows the cut surface of the petri dish under the combined effect test of perillaldehyde and citral on Candida albicans TIMM1768. The positional relationship between the paper disc, petri dish, medium, and vinyl tape is shown in the figure. Arrows indicate the diffusion of aldehydes from paper discs. (B) It is a photograph of a petri dish obtained as a result of the combined effect of perillaldehyde and citral on Candida albicans TIMM1768. The white part shows the part where Candida albicans TIMM1768 has grown, and the black part in the center shows that the growth is suppressed. FIG. 5 is a graph showing the results of measuring the area of the blocking circle obtained in FIG. 5 and plotting the points having an area of 3 cm ² (black circles) and the points having an area of 5 cm ² (black squares). The points where the FIC index is 0.583, 0.65, or 0.7 are indicated by arrows.

The anti-microbial active composition of the present invention is selected from a first aldehyde which is an acyclic terpene aldehyde and an aldehyde different from the terpene aldehyde, which is an acyclic aliphatic aldehyde, a terpene aldehyde and an aromatic aldehyde. It can be obtained by mixing with a second aldehyde.

In addition, the antimicrobial activity composition of the present invention can be included in the form of a kit together with an outer box, a container, a diluent, a gelling agent, and / or a description of a preparation method / administration method. When the antimicrobial active composition of the present invention is supplied as a kit, different constituents of the antimicrobial active composition may be packaged in separate containers and contained in one kit, or may be included in one kit. Even if only some of the components of the antimicrobial active composition (at least the first aldehyde and the second aldehyde) are included in the kit and another component is provided separately from the kit. Good. When the anti-microbial active composition of the present invention is supplied as a kit, the necessary constituents are preferably mixed immediately before use in order to obtain the anti-microbial active composition of the present invention. For example, the kit of the present invention comprises a first aldehyde which is an acyclic terpene aldehyde and an aldehyde different from the terpene aldehyde, which is selected from an acyclic aliphatic aldehyde, a terpene aldehyde and an aromatic aldehyde. It can be a kit for preparing an anti-microbial active composition comprising the second aldehyde.

In one aspect, the present invention comprises a first aldehyde which is an acyclic terpen aldehyde and an aldehyde different from the terpen aldehyde, which is selected from an acyclic aliphatic aldehyde, a terpen aldehyde and an aromatic aldehyde. It may be a method for preparing an anti-microbial active composition, which comprises mixing with a second aldehyde.

For example, the present invention comprises a first aldehyde which is an acyclic terpen aldehyde and a second aldehyde which is different from the terpen aldehyde and is selected from an acyclic aliphatic aldehyde, a terpen aldehyde and an aromatic aldehyde. The present invention relates to a method for suppressing the growth of microorganisms by applying or volatilizing (diffusing) an anti-microbial active composition containing an aldehyde. Preferably, the present invention comprises applying the antimicrobial active composition to the surface of the skin (eg, feet, hands, etc.) or to an object that comes into direct contact with the skin of a living body, such as shoes, socks, clothing, etc. It relates to a method of suppressing the growth of. Alternatively, the present invention relates to a method for suppressing the growth of microorganisms in the air, which comprises volatilizing and diffusing the antimicrobial active composition into a space (particularly, a closed space) such as a room or a car. In these methods, the method of suppressing the growth of microorganisms may be appropriately read as the method of deodorizing.

When the anti-microbial activity composition of the present invention is used by coating, it can be applied to a target place once to several times a day, if necessary. In addition, when the antimicrobial active composition of the present invention is used by volatilization / diffusion, it may be installed in a space for several hours to several months as necessary or by a method of promoting volatilization as necessary. it can.

The present invention will be described in more detail below with reference to examples, but this does not limit the scope of the present invention. References cited throughout the specification of the present application are incorporated herein by reference in their entirety.

(Example 1) Antibacterial effect of citral and perillaldehyde in combination against the odor-causing bacterium (Bacillus subtilis) Inoculate the SDA medium in a 35 mm petri dish with Bacillus subtilis (Bacillus subtilis), and put the two petri dishes in a closed box. It was installed in (inner diameter 150 × 100 × 98; combination 1.47L). 200 μl each of a citral alone solution, a perylaldehyde alone solution, or a mixed solution of citral and perylaldehyde diluted with 40% isopropanol was dropped onto the center of the filter paper (9 cm in diameter) attached to the lid of the closed box, and immediately covered. And sealed. After allowing to stand at 23 to 26 ° C. for 6 hours, the chalet was taken out from the closed box, the chalet was sealed with a parafilm, and the mixture was cultured at 37 ° C. for 20 hours. The number of Bacillus subtilis colonies appearing on the medium of the petri dish was counted and scored (Fig. 1A).

The antibacterial effect of the solution at the mixing ratio of each of the two components was displayed by the number of colonies on the agar medium by the checkerboard method, and was displayed as a score of 3 grades from 0 to 2 depending on the number of colonies. A plot of each concentration with a score of 0 is shown in FIG. 1B. FIC index = 0.5 (synergistic effect) at the value of the star mark was obtained. A photograph of the formed colonies is shown in FIG. 1C. As can be seen from the photograph, the effect of reducing not only the number of colonies but also the size of the colonies was observed at 1.25 mg / ml of perillaldehyde and 2.5 mg / ml of citral.

(Example 2) Effect of combined use of perylaldehyde and citral on the growth of tinea bacillus (Arthroderma vanbreusehemii TIMM2789) (sealed box)
After inoculating 10,000 minutes of Arthroderma vanbreusehemii TIMM2789 into SDA medium in a 35 mm petri dish, two petri dishes were placed in a closed box in the same manner as in Example 1. 200 μl each of citral alone solution, perillaldehyde alone solution, or citral and perillaldehyde mixed solution diluted with 40% isopropanol was dropped onto the center of the filter paper (9 cm in diameter) attached to the lid of the closed box, and the lid was closed immediately. Sealed. After allowing to stand at 23 to 26 ° C. for 16 hours, the chalet was taken out from the closed box, the chalet was sealed with a parafilm, and the mixture was cultured at 30 ° C. for 144 hours. The degree of growth of the hyphae of Trichophyton that appeared on the medium of the petri dish was scored and displayed (Fig. 2A).

The antifungal effect of the solution at the mixing ratio of each of the two components by the checkerboard method was displayed as a score of 4 grades from 0 to 3 depending on the degree of growth of ringworm hyphae on the agar medium. A plot of each concentration with a score of 0 is shown in FIG. 2B. FIC index = 0.375 (synergistic effect) at the value of the star mark was obtained.

(Example 3) Effect of combined use of various aldehydes including perillaldehyde and citral on the growth of Bacillus subtilis (Bacillus subtilis) (aromatogram)
Was applied Bacillus subtilis and (Bacillus subtilis) McFarland value 0.5 10-fold dilutions 100μl of (1 × ¹⁰ 8 CFU / ml) in Conradi stick to the standard agar surface of 90mm petri dish (10 ⁶ cells / dish ). Perillaldehyde serially diluted with 40% isopropanol was added dropwise to each of three thick filter papers placed on the surface of the agar medium as shown in FIG. 3A. Two similar petri dishes were prepared, and 50 μl of 40% isopropanol was added dropwise to the center of the first petri dish as a control, and 50 μl of citral diluent was added dropwise to the center of the second petri dish. All the solutions were dropped onto a thick filter paper as quickly as possible, covered with a petri dish, tightly sealed with vinyl tape, and then allowed to stand at 37 ° C. for 20 hours. The diameter of the inhibition circle on the agar medium after culturing was measured, and the difference in size between the case of using only the control perillaldehyde and the case of adding citral in the center was examined.

A photograph of the petri dish is shown in FIG. 3B. Table 1 shows the results of measuring the diameter of an inhibition circle having a size close to that of the fungus growth inhibition region on this photograph. When not only 40% isopropanol but also 5% citral was added to the center of the agar medium, the result was that the blocking circle due to the surrounding perylaldehyde rapidly increased. Similarly, combined effects were detected with cinnamaldehyde, cuminaldehyde, vanillin, citronellal and citral.

(Example 4) Effect of combined use of various aldehydes and citral on the growth of Candida albicans (TIMM1768) (aromatogram)
2 × 10 ⁶ Candida albicans TIMM1768 were applied to the surface of Sabouraud agar medium in a 90 mm chalet with a conlarge stick. Vanillin serially diluted with 40% isopropanol was added dropwise to each of the three locations shown in FIG. 4A on a thick filter paper placed on the surface of the agar medium in an amount of 50 μl. Three similar dishes were prepared, with 50 μl of 40% isopropanol as a control in the center of the first dish and 50 μl of 5% or 10% citral diluent in the center of the second and third dishes. Dropped. The operation was carried out as quickly as possible, all the solutions were dropped on thick filter paper, the petri dish was covered, and the surroundings were tightly sealed with vinyl tape. After standing culture at 37 ° C. for 20 hours, the diameter of the inhibition circle on the agar medium was measured, and the difference in size between the control vanillin and 40% isopropanol and the case where citral was added in the center was examined.

The results are shown in FIG. 4B. With the addition of vanillin, 5% and 10% citral, blocking circles were confirmed in both cases. The diameter of the circle was measured by superimposing a blocking circle having a size close to the growth blocking region of the fungus on the photograph shown in FIG. 4B. Table 2 shows the diameters of the blocking circles at the sites where vanillin of various concentrations was dropped. Compared with the case where only 40% isopropanol was added dropwise to the center of the agar medium, the results showed that the inhibition circle due to the addition of vanillin was larger when 5% and 10% citral were added. Therefore, it was shown that citral enhances the action of vanillin.

When a similar experiment was conducted on the combined action with citral using cinnamaldehyde or perylaldehyde instead of vanillin, a strong combined effect with citral was detected for both cinnamaldehyde and perylaldehyde.

(Example 5) Effect of combined use of perillaldehyde and citral on fungi (effect of combined use of volatile components)
By the cold weather method (aromatogram descent method), the antibacterial effect of the gas component against Candida, which belongs to fungi similar to ringworm, which is the causative agent of athlete's foot, was detected. As a material, citral (Nakarai 091-03 M5E4518) and perylaldehyde (Aldrich 21, 829-4 13812DN) were used. Candida albicans TIMM1768 was inoculated into Sabouraud-agar (SDA) medium in a petri dish (φ = 90 mm) so as to be 2 × 10 ⁶ per petri dish. Thick filter paper (φ = 8 mm; manufactured by ADVANTEC) was attached to the central part of the lid of the chalet with double-sided tape. Citral and perillaldehyde solutions are diluted with 40% isoamyl alcohol to a concentration of 0,2.5,5,7.5, 10,12.5, or 15w / v% of perillaldehyde and a concentration of 0,2.5. The mixture was mixed so as to be in combination with 5, 7.5, 10, 12.5, or 15 w / v% citral, and 50 μl of each was added dropwise to a thick filter paper attached to the shearlefta.

As shown in FIG. 5A, after dropping the solution onto a thick filter paper, the petri dish lid was quickly closed and sealed with vinyl tape. The cells were cultured at 37 ° C. for 20 hours, and the growth state of Candida and the circle of inhibition by aldehyde were photographed with a digital camera. From the captured image, the size of the blocking circle was measured with the area calculation software lenaraf220b, and determined as the strength of the antifungal effect. (At that time, the standard was that the diameter of the thick filter paper was 80 mm and the diameter of the petri dish was 90 mm.)

As shown in FIG. 5B, Candida grew as yeast on an agar medium and formed white colonies on the surface of the medium. When the concentrations of citral and perylaldehyde increased, a "blocking circle" was formed under the thick filter paper to which the solution was added, but the solution of perylaldehyde and citral alone formed a "blocking circle". A clearly larger blocking circle was formed than in the case.

From the result of taking a picture of the blocking circle of the culture chalet and measuring the area of the blocking circle by image analysis, the point where the area is 3 cm ² (the line where the lower points are plotted in the graph) and the point where it is 5 cm ² (graph) The result of plotting the middle and upper points) is shown in FIG. From this graph, the FIC index, which is an index of the combined effect, was obtained. As a result, values of 0.583, 0.65, and 0.7 were obtained, and it was clarified that a synergistic effect was obtained.

In addition, as a result of conducting a similar experiment using cuminaldehyde instead of perillaldehyde, the combined effect of cuminaldehyde and citral was confirmed (results not shown).

(Example 6) Combined antibacterial effect of various aldehydes and citral against fungi in liquid Regarding the combined antibacterial effect due to the volatile nature of the essential oil components confirmed in Examples 1 to 5, the combined antibacterial effect when directly sprayed with a spray or the like. In order to examine the effect, the combined effect on the cultured fungus in the solution was examined. As the strain used, Candida albicans TIMM1768 strain was used as a representative fungus. Bacteria were suspended in 1 / 3RPMI1640 medium (2.5% bovine serum free) was dispensed such that 10 ^4/100 [mu] l / well in 96-well plates min. Citral and decanal were diluted with DMSO to 10% w / w and further diluted with 1/3 RPMI1640 medium (v / v). 50 μl of each of the prepared decanal and citral solutions was added to the 96-well plate inoculated with the fungus to the final concentration shown in Table 3. All experiments were performed in triplicate (n = 3). In order to suppress the volatilization and diffusion of the active ingredient, a transparent plate seal was attached so as to cover the 96-well plate, the plate was further covered, and the cells were cultured at 37 ° C. for 20 hours. Under these culture conditions, the Candida albicans TIMM1768 strain grows from yeast to hyphal form, and the hyphae adhere to the bottom of the plastic, and the hyphae are quantified on the surface of the mycelium of a pigment (CV) called crystal violet. It has been found that it can be measured by the amount of adsorption. This CV staining method was used.

As shown in Table 3, Citral alone C.I. In order to suppress hyphal growth of albicans by 70% or more, a concentration of citral higher than the maximum concentration of 0.001% in the experiment conducted this time was required. In addition, Decanal alone C.I. A concentration of 0.006% was required to suppress hyphal growth of albicans by 70% or more. On the other hand, by using these two kinds of aldehydes in combination, the amount of citral was reduced to 0.0002% and the amount of decanal was reduced to 0.002%, and the same antibacterial effect could be obtained. That is, it was found that an antibacterial effect can be exhibited with a solution that is 5 times or more thinner for citral and 3 times thinner for decanal.

Similar tests were performed using octanal, nonanal, trimethylhexanal, hydroxycitroneral, perylaldehyde, benzaldehyde, cuminaldehyde, or vanillin instead of decanal, and all of them had the same antibacterial effect as decalal in combination with citral. Was obtained (results not shown).

Claims (5)

An anti-microbially active composition containing citral and perylaldehyde as active ingredients , the composition containing 1.25 to 25 mg / mL of perylaldehyde and 0.625 to 25 mg / mL of citral. Contains,
An antimicrobial active composition in which the microorganism is Bacillus subtilis, Trichophyton, or Candida . The anti-microbial active composition according to claim 1, which is a volatile preparation. The antimicrobial activity composition according to claim 1 or 2 , which has antimicrobial activity against microorganisms in the air. The antimicrobial active composition according to any one of claims 1 to 3 , which is a liquid preparation for spraying. The antimicrobial active composition according to any one of claims 1 to 4 , which is used for deodorizing.