JPH10305088A - Antimicrobial agent to which deodorizability is imparted - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an antimicrobial agent which consists of the components derived from plants as its effective component and to which deodorizability is imparted and an antimicrobial agent which is effective for food poisoning bacteria including enterophanogenic E. coli O-157. SOLUTION: This antimicrobial agent contains 'Smellnark (R)' and org. matter having a tropolone skeleton. An emulsifier is preferably incorporated further therein in addition to the 'Smellnark (R)' and the org. matter having the tropolone skeleton. The org. matter having the tropolone skeleton is preferably hinokitiol. Further, the antimicrobial agent is recommended to be made into gelatinous matter by adding a gelatinizing agent thereto.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an antibacterial agent having a deodorant property, comprising a plant-derived component as its active ingredient.

[0002]

2. Description of the Related Art Generally, a plant deodorant is obtained by extracting organic compounds from leaves, stems, etc. of a plant by a squeezing method, vacuum distillation, or the like, and mixing the organic compounds extracted from several types of plants. It has high safety because it contains plant-derived components as active ingredients. However, antibacterial properties are not generally imparted to such deodorants, and deodorant antibacterial agents having both antibacterial properties and deodorant properties are desired. By the way, in recent years,
At present, food poisoning due to food poisoning bacteria is occurring everywhere due to the spread of heating facilities in cooking facilities, etc. Especially, pathogenic Escherichia coli O157 is difficult to identify the source of infection. Has become.

[0003]

SUMMARY OF THE INVENTION Accordingly, an object of the present invention is to provide an antibacterial agent having a deodorant property, comprising a plant-derived component as its active ingredient. In particular, it is to provide an effective antibacterial agent against food poisoning bacteria such as pathogenic Escherichia coli O157.

[0004]

Means for Solving the Problems In order to solve the above problems, the present inventors have conducted intensive studies and found that a specific deodorant containing a plant extract extracted from many plants as an active ingredient has an antibacterial effect. By finding that an organic substance having a tropoton skeleton such as hinokitiol is blended, it has been found that both a high deodorizing effect and a high antibacterial effect can be obtained, and the present invention has been completed. That is, the antibacterial agent imparted with deodorant properties of the present invention is characterized by comprising Smernak (trade name) and an organic substance having a tropolone skeleton.
In addition, it is preferable that an emulsifier is further contained in addition to the organic substance having a trade name of Smernak and a tropolone skeleton. Further, as the organic substance having the tropolone skeleton, hinokitiol is preferable. It is preferable to add a gelling agent to form a gel.

[0005]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments of the present invention will be specifically described below. In the present invention, trade name Smernak is a deodorant manufactured by Tokai Kosan Co., Ltd., cedar, cypress, fir, nettle, camphor, black pine, red pine, spruce, persimmon, kumasasa, mugwort,
It is an extract that extracts about 180 kinds of organic compounds from 35 kinds of plants mainly composed of tea, birch, perilla aloe, sunshaw, ama tea slurries, etc. by using a unique squeezing device and vacuum distillation device, and blends them. . Such an extract is water-soluble and has a high deodorizing effect.

Examples of the organic substance having a tropolone skeleton include hinokitiol (β-thyaprisin), β-drabulin, and derivatives thereof. Here, hinokitiol is an oil-soluble crystalline substance present in Aomori-grown hiba oil, Taiwan hinoki oil, Western Red Cedar oil and the like, and has strong antibacterial properties. Β-Drabulin is contained in Aomori Hiba oil and has an antibacterial property equivalent to that of Hinokitiol.

[0007] The above-mentioned trade name Smernak is water-soluble,
Since hinokitiol as an organic substance having a tropolone skeleton is oil-soluble, an emulsifier (surfactant) is added to uniformly mix them. Such an emulsifier is not particularly limited as long as both can be uniformly mixed and emulsified, such as castor oil. Further, when an organic acid, an organic acid salt or an organic acid ester is added, the emulsifiability is increased. Examples of such organic acids include citric acid, sodium citrate, triethyl citrate and the like. Water is used as the solvent, but alcohol can also be added because hinokitiol has alcohol solubility. The water used here is preferably ion-exchanged water that does not contain impurities such as chlorine and minerals.

Although the mixing ratio of the above components varies depending on the type of components used, when hinokitiol is used as an organic substance having a tropolone skeleton, Smernak 0.3 to
About 30% by weight, about 0.1 to 2% by weight of hinokitiol, about 0.5 to 10% by weight of an emulsifier, and about 0.5 to 5% by weight of organic acids. If the amount of Smelnark is too small, the deodorizing effect becomes insufficient, and if it is too large, the cost increases. If the amount of hinokitiol is too small, a sufficient antibacterial effect cannot be obtained, and if it is too large, the cost increases. If the addition amount of the emulsifier and the organic acids is too small, the emulsification becomes insufficient and the hinokitiol crystallizes and precipitates,
On the other hand, if it is too large, the emulsifier and the like become cloudy and the amount of precipitate increases.

As a form of use, it can be used as an aqueous solution (liquid substance). However, for use in refrigerators, cars, indoors, etc., it is preferable to put it in a container as a gel to volatilize the active ingredient. It is convenient. In order to obtain a gel, a water-absorbing resin may be added. Active ingredients such as smernak and hinokitiol have volatility, and by making them into a gel, the surface area is increased to promote the volatilization of the active ingredients, thereby improving the deodorizing effect and the antibacterial effect. When left in the air as a gel, the water in the water-absorbent resin evaporates. At this time, the water evaporates with the active ingredient adsorbed on the surface of the resin.

When used as a gel, the water-absorbing resin is blended in an amount of about 4 to 10% by weight based on the whole gel. If the compounding ratio is too small, the surface area becomes insufficient and the volatility of the active ingredient deteriorates, and if it is too large, the volatility improves, but the bulk specific gravity becomes small, and a large container is used to contain the same amount of the effective component. I have to.

[0011]

The present invention will be described more specifically with reference to the following examples. These examples do not limit the scope of the claims. [Antibacterial effect test] The antibacterial effect was tested for each of the brand name "Smernark" alone, catechin alone, hinokitiol alone, and a mixture of "Smernark" and hinokitiol.

EXPERIMENTAL EXAMPLE 1 A solution having a predetermined concentration of "Smernark" (trade name) was used as a test solution, and the test solution was inoculated with a bacterial solution of Escherichia coli (serotype O157: H7), allowed to act at room temperature, and tested with time. The presence or absence of surviving bacteria in the liquid was examined.

Escherichia coli, serotype O157 as a test strain:
H7, non-verotoxin-producing strain (Escherichia coli ATCC 4388
8), a test strain pre-cultured at 37 ± 1 ° C. for 16 to 24 hours in an NA medium (ordinary agar medium (manufactured by Eiken Chemical Co., Ltd.)) was again inoculated into the NA medium, and then inoculated at 37 ± 1 ° C., 16 to The cells cultured for 20 hours were uniformly dispersed in physiological saline, and a bacterial solution was prepared so that the number of cells per ml was about 10 ⁸ . As a test solution, a 10, 5, 2.5% (V / V) solution of the sample was prepared as a sample (trade name “Smernark”) undiluted solution and used as a test solution. Then, 1m of bacterial solution is added to 10ml of test solution.
1 and inoculated at room temperature, and after 0.5, 1, 3 and 6 hours, one loopful of NB medium (Normal broth with 0.2% meat extract (manufactured by Eiken Chemical Co., Ltd.)) Was inoculated and cultured (cultured at 37 ° C. for 2 days) to check for growth. Table 1 shows the results. The sign + in the table indicates that there is growth, and the sign-indicates that there is no growth.

[0014]

[Table 1] Table 1 Bactericidal effect test results (according to Japan Food Research Laboratories) Test bacteria Sample concentration Growth % (V / V) 30 minutes 1 hour 3 hours 6 hours later Escherichia coli 100 (stock solution) +---(serotype 10 + + + + O157: H7) 5 + + + + + + + + + +

Experimental Example 2 Catechin was used as a sample, and catechin was purified with purified water.
The test was conducted in the same manner as in Experimental Example 1 except that 0, 0.5, and 0.1% (W / V) solutions were prepared and used as test solutions. Table 2 shows the results.
Shown in

[0016]

[Table 2] Table 2 Bactericidal effect test results (according to Japan Food Research Laboratories) Test bacteria Sample concentration Growth % (V / V) 30 minutes 1 hour 3 hours 6 hours later E. coli 1.0 ++++++ (serotype 0.5 ++++ O157: H7) 0.1 ++++

EXPERIMENTAL EXAMPLE 3 Hinokitiol was used as a sample, and 5
% (V / V) 1.0, 0.1, 0.0 with ethanol solution
A test was conducted in the same manner as in Experimental Example 1 except that 1 and 0% (W / V) solutions were prepared and used as test solutions. Table 3 shows the results.

[0018]

[Table 3] Table 3 Bactericidal effect test results (according to Japan Food Research Laboratories) Test bacteria Sample concentration % (V / V) 15 minutes 30 minutes 1 hour 3 hours 6 hours later Escherichia coli 1.0 + +---(serotype 0.1 + + + + + O157: H7) 0.01 + + + + + 0 + + ++++

Experimental Example 4 Sample 1) was trade name "Smecnal" and sample 2) was hinokitiol, and 2.5% (V / V) of sample 1).
V) A solution obtained by adding about 1% of the sample 2) to the solution is used as a test solution, and bacterial solutions of various bacteria are inoculated into the test solution and allowed to act at room temperature, and the presence or absence of surviving bacteria in the test solution is determined over time. Examined.

As test strains (1) Escherichia coli, serotype O1
57: H7, non-verotoxin-producing strain (Escherichia coli ATC
C 43888), (2) Escherichia coli IFO 397
2), (3) Staphylococcus aureus
IFO 12732), (4) Salmonella enterit
idis IFO 3313), (5) Vibrio parahaemolyticus (Vibrio parah)
aemolyticus IFO 12711) and E. coli (O157: H
7) For Escherichia coli, Staphylococcus aureus and Salmonella, a test strain pre-cultured at 37 ± 1 ° C. for 16 to 24 hours in NA medium (normal agar medium (manufactured by Eiken Chemical Co., Ltd.)) was inoculated again into NA medium. The cells cultured at 37 ± 1 ° C. for 16 to 20 hours were uniformly dispersed in physiological saline to prepare a bacterial solution such that the number of bacteria per ml was about 10 ⁸ . In addition, for Vibrio parahaemolyticus, an NA medium containing 3% NaCl (NA medium containing 3% sodium chloride) at 37 ± 1 ° C.
The test strain, which had been pre-cultured for 24 hours, was again inoculated into a 3% NaCl-added NA medium, and the cells cultured at 37 ± 1 ° C. for 16 to 20 hours were uniformly dispersed in a 3% sodium chloride solution.
A bacterial solution was prepared so that the number of bacteria per cell was about 10 ⁸ .
A test solution was prepared by adding 4 ml of a 20% (W / V) ethanol solution of sample 2) to 80 ml of a 2.5% (V / V) solution of sample 1).

Then, 10 ml of the test solution was inoculated with 1 ml of the bacterial solution, allowed to act at room temperature, and after 30 minutes, 1, 3, and 6 hours, one loopful was added to an NB medium (normal broth containing 0.2% meat extract). (Manufactured by Eiken Chemical Co., Ltd.)
C. for 2 days) to check for growth. For Vibrio parahaemolyticus, an NB medium supplemented with 3% NaCl (NB medium supplemented with 3% sodium chloride) was used instead of the NB medium. Table 4 shows the results.

[0022]

[Table 4] Table 4 Bactericidal effect test results (according to Japan Food Research Laboratories) Test Bacterial growth Yes / No 30 minutes 1 hour 3 hours 6 hours later E. coli (O157: H7)----E. coli----Staphylococcus aureus----Salmonella ----Vibrio parahaemolyticus----

As can be seen from the above experimental results, the use of Smernak alone against Escherichia coli (O157: H7) has a bactericidal effect in the stock solution, but no effect upon dilution. Catechin has no bactericidal effect. Hinokitiol has a bactericidal effect. The mixture of smectar and hinokitiol has a stronger bactericidal effect against Escherichia coli (O157: H7) and also against other bacteria (Escherichia coli, Staphylococcus aureus, Salmonella, Vibrio parahaemolyticus). A strong bactericidal effect is observed.

[Production of gel-like material ] Gel-like material 1 Smernak 2.5% by weight, hinokitiol 1.0% by weight, undenatured alcohol (ethanol) 5% by weight, polyoxyethylene hydrogenated castor oil (Nippon Surfactant Industries, Ltd.) as an emulsifier Product name NIKKOL HC
O-50) 2.0% by weight and polyoxyethylene hydrogenated castor oil (trade name NIKKOL HCO-60, manufactured by Nippon Surfactant Industries, Ltd.) 2.0% by weight, citric acid 0.3% by weight, sodium citrate 0.2 % By weight, water-absorbing resin (trade name: KI gel 201K, manufactured by Kuraray) 4.0
Weight%, and the mixing ratio so that the balance becomes ion-exchanged water,
Hinokitiol was dissolved in undenatured alcohol, and hinokitiol, smernak and other components dissolved in the alcohol were dissolved in ion-exchanged water and mixed uniformly. Next, a water-absorbent resin was added to and mixed with this mixed solution to obtain a pale yellow translucent gel-like substance 1.

Gel 2 The amount of the unmodified alcohol (ethanol) was adjusted to 20.0%.
The same treatment as described in Gel 1 above was carried out except that the amount was changed to% by weight, to obtain a pale yellow translucent gel 2. The gel 1 and the gel 2 contain alcohol and thus have an alcohol odor. However, when the gel is left for 2 to 3 days, the alcohol volatilizes and the alcohol odor disappears.

Gel 3 : 2.5% by weight of Smernak, 1.0% by weight of hinokitiol, 3.0% by weight of triethyl citrate, polyoxyethylene hydrogenated castor oil as emulsifier (product name NIKKOL HCO-, manufactured by Nippon Surfactant Industries, Ltd.) 5
0) 1.0% by weight, water-absorbent resin (trade name K manufactured by Kuraray)
I-gel 201K) 4.0 wt%, the balance being ion-exchanged water, hinokitiol, triethyl citrate, and NIKKOL HCO-50, and a portion of ion-exchanged water was added thereto and emulsified with a mixer. did. Next, the remaining ion-exchanged water and Smernak were added and mixed uniformly. Next, a water-absorbent resin was added to and mixed with this mixture to obtain a pale yellow translucent gel-like substance 3. This gel 3 was substantially odorless.

[Test of antibacterial effect of gel-like material] A gel-like material 3 was used as a sample 1) and a gel-like material 2 was used as a sample 2).
As shown in FIG. 1, a gel material (specimen) 10 was packed in a container 11 and a plate medium 12 coated with bacterial solutions of various bacteria.
Into a sealed container (about 2.7 liters)
After culturing at 10 ° C. and 35 ° C. in No. 3, the viable cell count was measured.

The same test strain as that used in Experimental Example 4 above was used, and Escherichia coli (O157: H7),
For E. coli, Staphylococcus aureus and Salmonella, NA
37 ± in the medium (normal agar medium (Eiken Chemical Co., Ltd.))
The test strain pre-cultured at 1 ° C. for 16 to 24 hours was again inoculated into an NA medium, and the cells cultured at 37 ± 1 ° C. for 16 to 20 hours were uniformly dispersed in a phosphate buffer. A bacterial solution was prepared to be about 10 ³ . As for Vibrio parahaemolyticus, a medium containing 3% NaCl (NA medium containing 3% sodium chloride) at 37 ± 1 ° C., 16 to 24 ° C.
The test strain pre-cultured for 3 hours is again inoculated into 3% NaCl-added NA medium, and the cells cultured at 37 ± 1 ° C. for 16 to 20 hours are uniformly dispersed in a 3% sodium chloride solution. A bacterial solution was prepared to be 10 ³ .

For Escherichia coli (O157: H7), Escherichia coli, Staphylococcus aureus and Salmonella, 0.1 ml of the bacterial solution was spread on an SA plate medium (standard agar medium (manufactured by Eiken Chemical Co., Ltd.)) 12. Closed container 13 with 10
And incubated at 10 ° C for 7 days and at 35 ° C for 2 days.
The number of viable bacteria was measured. For Vibrio parahaemolyticus, an SA plate medium supplemented with 3% NaCl (SA medium supplemented with 3% sodium chloride) was used instead of the SA plate medium. In addition, those cultured without the specimen 10 (control) were also tested in the same manner. Table 5 shows the results. In Table 5, the symbol "-" indicates that the cells did not grow.

[0030]

[Table 5] Table 5 Antibacterial activity test results (according to Japan Food Research Laboratories) Test bacteria Number of viable bacteria per plate 35 ° C culture 10 ° C culture colon bacterium Sample 1) 0 (O157: H7) Sample 2) 0 0 Control 91 179 samples 1) 0 0 E. coli sample 2) 0 0 Control 49 141 Sample 1) 0-Staphylococcus aureus Sample 2) 0- Control 96 - Sample 1) 0 0 Salmonella Sample 2) 0 0 Vs. irradiation 95 76 sample 1) 0 - Vibrio parahaemolyticus sample 2) 0 - versus irradiation 37 -

From the results shown in Table 5, it can be seen that even when the antibacterial agent of the present invention is made into a gel, the active ingredient volatilized from the gel shows a high antibacterial effect on various bacteria. In addition, about Staphylococcus aureus and Vibrio parahaemolyticus, 10
At ℃, the cells did not grow, indicating that the cells were weak to low temperatures.

[Test for deodorizing effect of gel-like material] 180 g of gel-like material 1 (G1) and 170 of gel-like material 2 (G2) 170
g in a container having a diameter of 72 mm and a height of 55 mm,
With the upper part of the container opened, it was installed in various places to test the deodorizing effect. Table 6 shows the results.

[0033]

Table 6 specimens G1 G1 G1 G1 G1 G2 G2 location toilet Refrigerator Kitchen-room office interior toilet refrigerator average size 220 l 8 tatami mats 14.4 tatami mats 283 l Number of arrangements 2 6 2 5 2 1 3 Average installation days (days) 32.5 35.3 32.5 26.6 25.5 26 23.7 Average reduction / day (g) 1.8 2.19 2.72 3.82 4.16 0.92 2.31 Average temperature (℃) 10 5 16.5 22.8 25 10 5 Average humidity (%) 50 53 Odor check good good good good good good good freshness retention check good good

From the results shown in Table 6, it can be seen that the deodorizing effect is good at any installation location. In addition, when installed in a refrigerator, freshness of fresh food such as vegetables placed in the refrigerator is 20 to 4 times higher than when no gel-like material is installed.
It was held for about 0% longer. This is probably because the antibacterial effect was exhibited.

[Analysis of Content of Heavy Metals, Poisons, etc.] The analysis of the content of heavy metals, poisons, etc. was carried out on the liquid obtained by the same treatment as described for the gel 3 except that the gelling agent was not added. The test was performed. Table 7 shows the results. Table 7
It can be seen from the results that no heavy metals or toxic substances were detected from the antibacterial agent of the present invention.

[0036]

[Table 7] Table 7 Analytical test results (according to Japan Food Research Laboratories ) Analytical test items Results Detection limit analysis method EPN not detected 0.05 ppm gas chromatographic method Parathion not detected 0.05 ppm gas chromatographic method Methyl dimeton detection Not detected 0.05 ppm Gas chromatographic method Methyl parathion Not detected 0.05 ppm Gas chromatographic method Arsenic (as As) Not detected 0.1 ppm DDTC-Ag Spectrophotometric method Lead Not detected 0.05 ppm Atomic spectrophotometric method Cadmium Not detected 0.01 ppm Atomic absorption method Photometric method Total mercury not detected 0.01 ppm reduced vapor atomic absorption spectrophotometric method Total chromium not detected 0.5 ppm diphenylcarbazide spectrophotometric method Cyan Not detected 0.1 ppm pyridinepyrazolone spectrophotometric method

[Acute Oral Toxicity Test in Mice] The same liquid substance used in the above-mentioned analysis test for heavy metals, toxic substances, etc. was used as a sample, and an acute oral toxicity test in mice was carried out in accordance with the OECD Guideline for Chemical Toxicity Test (1987). A toxicity test (limit test) was performed at the Japan Food Research Laboratories.

As test animals, 4-week-old ICR male and female mice were purchased from Japan SLC Co., Ltd., and were preliminarily reared for about one week to confirm that there was no abnormality in their health. Five test animals were housed in polycarbonate cages, each being housed in a breeding room set at a room temperature of 23 ± 2 ° C. and an illumination time of 12 hours / day. Feed (mouse / rat solid feed (Lab MR Stock, manufactured by Nihon Nosan Kogyo Co., Ltd.)) and drinking water (tap water) were allowed to be taken freely.

The test and control groups each used 10 males and 10 females, and the test animals were fasted for about 4 hours before administration. After measuring the body weight, in the test group, 20 ml /
Specimens were given a single oral gavage at a dose of kg using a gastric probe. The control group received 0.6 ml of purified water for males and 0.5 ml for females in the same manner. The observation period was 14 days, and the administration was performed frequently on the administration day and once a day from the next day. Body weight was measured every week after administration, and comparison between groups was performed at a significance level of 5% using t-samples. All animals were necropsied at the end of the study period.

As a result of the above test, no deaths were observed during the observation period for both males and females. As a clinical symptom, in the test group, a decrease in locomotor activity was observed in all cases from a few minutes after administration in both males and females, writhing reaction and abnormal body posture (prone position) were observed in 2 males and 1 female. Was. In addition, staggered walking was also observed in four males and four females. The agony reaction was 30 minutes after administration,
Other symptoms generally recovered after 5 hours, and 1 hour after administration.
No abnormalities were seen after the day. In the control group, no abnormality was observed throughout the observation period. Table 8 shows the results of measuring the body weight at 1 week and 2 weeks after administration as a change in body weight.
The body weight was represented by a mean value ± standard deviation (unit: g). The number of animals is shown in parentheses. This weight measurement showed no difference in weight gain between the test group and the control group in both sexes. Necropsy findings showed no major organ abnormalities in each group in both sexes at necropsy after the observation period.

According to the OECD Guidelines for Chemical Toxicity Test (1987), when the sample is a liquid, the dose is 2 per 100 g of body weight.
ml (20 ml / kg) should not be exceeded, and no deaths were observed at the highest dose that could be administered in this study, and no abnormalities were seen at necropsy. Therefore, it was confirmed that the lethal dose of a single oral administration of the sample to mice was 20 ml / kg or more for both males and females.

[0042]

Table 8 Weight change administration group Before administration After administration 7 days 14 days test group 27.1 ± 0.9 (10) 31.3 ± 1.5 (10) 34.8 ± 2.0 (10) male control group 27.4 ± 1.0 (10) 32.6 ± 1.6 (10) 36.0 ± 2.0 (10) test group 22.9 ± 0.6 (10) 24.7 ± 1.1 (10) 28.2 ± 1.7 (10) Female control group 22.5 ± 0.5 (10) 25.6 ± 1.1 (10) 28.2 ± 1.8 (10)

[0043]

As described above, according to the antibacterial agent imparted with deodorizing properties of the present invention, a plant-derived component is used as its active ingredient, so that it is highly safe and has both deodorizing properties and antibacterial effects. be able to. Further, by making the antibacterial agent imparted with deodorant properties of the present invention a gel, it is possible to promote the volatilization of the active ingredient and enhance the deodorant and antibacterial effects. In particular, a high antibacterial effect is obtained against food poisoning bacteria such as pathogenic Escherichia coli O157.

[Brief description of the drawings]

FIG. 1 is an explanatory diagram of a method for testing an antibacterial effect of a gel.

[Explanation of symbols]

 Reference Signs List 10 Gel-like substance (sample) 11 Container 12 Plate medium 13 Sealed container

Claims (4)

[Claims] 1. An antibacterial agent having deodorant properties, comprising Smernak (trade name) and an organic substance having a tropolone skeleton. 2. The deodorant-providing antibacterial agent according to claim 1, further comprising an emulsifier in addition to the organic substance having a trade name of Smernaq and a tropolone skeleton. 3. The antibacterial agent imparted with deodorant properties according to claim 1, wherein the organic substance having a tropolone skeleton is hinokitiol. 4. The antibacterial agent having a deodorant property according to claim 1, wherein a gelling agent is further added to form a gel.