JPH0640834A - Spray comprising natural antimicrobial substance and its production - Google Patents

Abstract

PURPOSE:To obtain a spray comprising a natural antimicrobial substance, controlling microorganisms existing in a target to be regulated by spraying the substance in a mist state on the target to be regulated. CONSTITUTION:Seeds are taken from Citrus paradisi (Japanese name: grapefruit) and ground. A liquid is extracted from the ground seeds and diluted with water to prepare a natural antimicrobial substance, which is packed into a container of spray. The ground extracted solution comprises, lipid, glucide, protein, water, etc., contains no arsine, no lead, no cadmium, etc., harmful to human body at all, is negative on minimum to ordinary bacteria and a group of Escherichia. The spray is useful for controlling microorganisms in rooms, automobiles, food processing spots, kitchens, hospitals, etc., in extremely high safety.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a spray composed of a natural antibacterial substance, and more specifically, by spraying it in the form of mist on an antibacterial target such as indoors, cars, food processing plants, kitchens and hospitals. The present invention relates to a spray composed of a natural bacteriostatic substance that sterilizes bacteria present in a bacteriostatic target.

[0002]

2. Description of the Related Art As is well known, indoors, cars, food processing plants,
Various bacteria exist in kitchens, hospitals, etc. Therefore,
Sprays composed of antibacterial substances are often used to control the growth of bacteria.

In this case, it is safe for the person who handles the spray, for the bactericidal target to which the spray is applied, and for the environment around it, it has no side effects, and it is easy to handle. There is a need for a spray of antibacterial material that is inexpensive.

However, in reality, there are no good antibacterial substances that satisfy the above requirements, and the antibacterial substances that have been used in the past have been so-called chemical compounds.

[0005]

According to the above-mentioned prior art, it has been recognized that the above-mentioned chemical-based antibacterial substances have the effect of controlling the bacteria present in the subject to be controlled.

However, chemical antibacterial substances are not decomposable and are not easily decomposed, which may cause a danger to the person handling the spray, the antibacterial target to which the spray is applied, and the environment around it. There was a problem that there is a property.

Further, since there is a possibility of danger,
There is a problem in that it becomes necessary to set the usage standard such as the concentration of the antibacterial substance, the usage time, and the usage target, which makes the handling extremely difficult.

Further, there has been a problem that the color may change in appearance or the like may be affected depending on the type of chemical system used by the applied bacteriostatic target.

[0009]

[Purpose] Therefore, the object of the present invention is not only to control the bacteria present in the bacteriostatic target, but especially to those who handle the spray, the bactericidal target to which the spray is applied, and the environment around it. To provide a spray consisting of a natural antibacterial substance which is safe against and easy to handle.

[0010]

[Means for solving the problem]

In order to achieve the above object, the present invention has the following technical means. That is, the present invention is a spray composed of a natural bacteriostatic substance that sterilizes the bacteria present in the bacteriostatic target, and this natural bacteriostatic substance is an aqueous solution of an extract extracted from crushed citrus paradecy seeds. It is characterized by being. This citrus paradisi is a scientific name and is written in Roman letters as "Citrus Paradisi", and as expressed by this scientific name, the following is included. That is, Japanese name Grapefruit, Pomelo, US, UK Grapefru
It, Pomelo, Brazil Pomelo, Spain Pomelo and so on.

The composition of the crushed extract of citrus paradecy seeds itself consists of lipids, sugars, proteins, water and the like. And in this, arsenic (A
s2O3), lead, cadmium, etc. are not included at all. In addition, general bacteria and coliforms are negative or minimal.

In addition, in the toxicity test of the aqueous solution of the crushed extract of citrus paradecy seeds, that is, the test of oral administration to animals such as rats, no poisoning symptoms and death cases were found, and moreover, there was no change in weight. Normal growth was observed without any occurrence, and it was confirmed to be nontoxic.

As described above, since the bacteriostatic substance is a natural product, it has been confirmed to be harmless. The analytical values and test results are clarified in the following examples.

The above-mentioned natural bacteriostatic substance is contained in a spray container. A pressure pump is attached to the container in order to spray the natural bacteriostatic substance from the inside of the container to the outside in the form of mist. Further, in order to spray the natural bacteriostatic substance under self-pressure, it is possible to mix a filling gas such as LP gas, dimethyl ether, carbon dioxide gas or nitrogen gas into the natural bacteriostatic substance. Incidentally, in order to enhance the volatility of the natural bacteriostatic substance after spraying it on the bacteriostatic target, it is possible to mix denatured alcohol with the natural bacteriostatic substance.

Further, the present invention is a method for producing a spray comprising a natural bacteriostatic substance for bacteriostatically controlling the bacteria present in the bacteriostatic target. The natural bacteriostatic substance takes out seeds from Citrus paradecy and then takes them out. The seeds are crushed, and the liquid is extracted from the crushed seeds and diluted with water.

As the treatment operation for extracting the liquid from the crushed citrus paradecy seeds in the above, any method such as a heating distillation method, a hydrolysis method, a solvent extraction method and a pressing method may be used.

According to the above-mentioned manufacturing method, citrus paradeci seeds can be obtained in large quantities at low cost.
It is easy and cheap to produce using this material,
Furthermore, it can be easily applied to antibacterial subjects without paying special attention. The food itself is safe and odorless to the human body, and has no restrictions on usage or concentration.

[0018]

The preferred embodiments of the present invention will now be described in detail with reference to the accompanying drawings. First, seeds of citrus paradecy were crushed and subjected to a hydrolysis method to obtain an extract therein. The analysis results are shown in Tables 1, 2 and 3. Table 1
Is the composition analysis result, Table 2 is the toxic component analysis result, and Table 3 is the bacterial analysis result.

[Table 1]

[Table 2]

[Table 3]

The extract thus confirmed to be safe was diluted 300 times with dilution water. Of course, this dilution ratio can be selected in various other ways. Incidentally, this natural bacteriostatic substance was further subjected to an acute toxicity test in order to confirm the safety. The results of the acute toxicity test are shown below.

1. Test animals and administration route, ddy-N mouse male and female produced by Japan Medical Science Animal Materials Research Institute Co., Ltd. were introduced at about 4 weeks of age, and then preliminarily reared for 1 week to show no abnormality in health. After confirming that, it was put into the test at about 5 weeks of age. The body weight at the start of the test was 24-26 grams for males and 22 grams for females. These test animals were orally administered.

2. Room temperature, 22 degrees Celsius plus or minus 2
In and out.

3. Test liquid (natural antibacterial substance) preparation method,
The sample was diluted with distilled water for injection to give 5.79, 6.94,
8.33 and 10% (W / V) aqueous solutions were prepared and used as test solutions.

4. Administration method of test solution, etc. a. Administration method: The mice to be used were fasted for 13 hours from 10 pm on the previous day to 11:00 am on the next day, and then the test solution was forcibly orally administered once using a stomach stomach. B. LD50 value calculation method, probit method, c. Dose of the sample, by administering the test liquid to each group 20ml per kilogram body weight,
The sample dose was 2,000 milligrams per kilogram as the upper limit, and 4 levels were administered to both males and females with a common ratio of 1 and 2. D. Number of animals per test group, 5 males and 5 females.

5. The test results and the results are shown in Table 4.

[Table 4]

6. Intoxication symptoms, no abnormalities were observed immediately after administration in either male or female in any of the administration groups, and then 2
Even during the observations carried out over a week, no remarkable symptom or death was observed.
All mice started feeding immediately after the administration.

7. Autopsy findings, for all survivors,
At the end of the test, the animals were sacrificed and necropsied visually, but no abnormality was observed in the major organs in either male or female in any of the administration groups.

8. Discussion, OECD Guidelines (19
According to the acute toxicity test method of April 11, 1986, etc., the maximum dose of the test substance for mice was 2 per kilogram.
It is specified as 000 milligrams. In this test, the sample was diluted with distilled water for injection to give 5.79, 6.94, 8.
33 and 10% (W / V) aqueous solutions were prepared and administered to mice at 20 ml per kilogram to obtain 2,000 per kilogram as a sample.
Four levels were administered to both males and females at a common ratio of 1 and 2, with 0 mg as the upper limit. As a result, no poisoning symptoms were observed in both males and females after administration of the test solution, which was considered to be due to the test solution administration, and no deaths occurred at any dose level.
Further, the growth during the test period is as shown in Table 2,
All were considered to show almost normal development. Therefore, the LD50 value of oral administration of the sample was estimated to be 2,000 mg / kg or more.

9. Table 5 shows changes in body weight of test animals.
The results are shown in Table 6 and Table 7. In addition, Table 6 shows the weight of each individual male, and Table 7 shows the weight of each female individual, and the unit is gram.

[Table 5]

[Table 6]

[Table 7]

Next, Example 1 of the spray comprising the natural antibacterial substance having the above characteristics will be described. The natural bacteriostatic substance is an extract obtained by crushing grapefruit seeds, diluted 300 times with water, and further diluted 30 to 3,000 times with water. Is contained in a spray container. And a pressure pump is attached to the container,
The pressurizing pump sprays the above-mentioned natural bacteriostatic substance from the inside of the container to the outside in the form of mist.

A second embodiment will be described. In the spray container of this Example 2, 60% of the natural bacteriostatic substance of the above Example 1 and LP gas, dimethyl ether, carbon dioxide gas,
40% of a filling gas such as a mixed gas of nitrogen gas and LP gas and dimethyl ether is mixed, and the natural bactericidal substance is sprayed in a mist form by the self-pressure of the filling gas.

Next, a third embodiment will be described. In the spray container of the third embodiment, 40% of the natural bacteriostatic substance of the first embodiment, 40% of the filling gas of the second embodiment, and 20% of denatured alcohol are mixed, and the self-pressure of the filling gas. Thus, the natural bacteriostatic substance and the denatured alcohol are sprayed in the form of mist, and the volatility becomes high after the bacteriostatic target is sprayed by the denatured alcohol.

Next, Test Example 1 using a natural bacteriostatic substance will be described. This Test Example 1 is a measurement of the minimum inhibitory concentration of a natural bacteriostatic substance against various microorganisms.

The test method of Test Example 1 will be described.
First, the test strains are as follows. Bacteria (excluding lactic acid bacteria); Escherichia coli IFO
3301, Pseudomonas
aeruginosa) IFO13275, Pseudomonas fluorescens IAH12
001, Nicrococcus lute
us) ATCC9341, Staphylococcus aureus IFO12732, Bacillus subtilis ATCC
6633, Bacillus cereus
) ATCC 19637, Bacillus megaterium (B
acillus megaterium) IFO12108, lactic acid bacterium; Lactobacillus brevis
IFO3345, Lactobacillus plantarum IFO3070, Leuconostoc mesenter
oides) IFO3426, Streptococcus faecalis IFO12964, yeast; Candida albicans I
FO1594, Hansenula Anomala Bar. Anomala (Hansenula anomala var.anomala) IFO1021
3. Saccharomyces ce
revisiae) IFO1950, Zygosaccharomyces rouxii IFO187
6, mold; Aspergillus niger
ger) IFO4407, Cladosporium Cladosporium IFO6
348, Penicillium citr
inum) IFO7784, Rhizopus stonifa (Rizo
pus stolonifer) IFO5411.

The culture medium for enrichment is as follows. Bacteria ...
Mueller Hinton Broth
(Difco)), lactic acid bacteria ... Heart Infusion Broth (Difco), yeast ...
5% agar-added glucose peptone medium (Nissui Pharmaceutical), mold
・ Potato dextrose agar medium (Eiken Chemical).

The medium for measuring sensitivity is as follows. Bacteria: Mueller Hinton Medium
nton Medium (Difco)), lactic acid bacteria ··· Heart Infusion Agar (Difco), mold, yeast · · 1.5% agar-added glucose peptone medium (Nissui Pharmaceutical).

The plate for sensitivity measurement was prepared by adding an arbitrary amount of the test sample to a medium for sensitivity measurement kept at 50 to 60 ° C. after sterilization and lysis, thoroughly mixing, dispensing into a petri dish and solidifying. The plate was used for sensitivity measurement.

The preparation of the bacterial solution for inoculation is as follows. Bacteria: Inoculate the subcultured test bacterium into the enrichment medium, and incubate at 35 degrees Celsius for 18 to 20 hours, then use the enrichment medium so that the number of bacteria is 10 6 per milliliter. It was diluted to obtain a bacterial solution for inoculation. However, Pseudomonas fluorescens was cultured at 30 degrees Celsius. Mold ··· Subcultured test strain was inoculated into the culture medium for enrichment and cultured at 25 degrees Celsius for 7 days, and the spores (conidia) formed were sterilized with 0.005% dioctyl sodium sulfosuccinate solution to about 10 The cells were floated so as to be a multiplicity of cells and used as a bacterial solution for inoculation. Yeast ··· Subcultured test bacteria are inoculated into the culture medium for enrichment, cultured at 25 degrees Celsius for 2 days, and then suspended in sterilized physiological saline so that the number of cells is about 10 6 It was a liquid.

The culture was carried out by using a nichrome wire loop (internal diameter of about 1 mm) on the susceptibility measuring plate to inoculate the bacterial solution.
Streaks about 1 to 2 centimeters, bacteria is 35 degrees Celsius
Molds were cultivated at ˜37 ° C. for 18 to 20 hours, fungi at 25 ° C. for 7 days, and yeast at 25 ° C. for 2 days. However, Pseudomonas fluoresc
ens) was cultured at 30 degrees Celsius.

For the determination, the minimum concentration at which the growth was inhibited after culturing was defined as the minimum inhibitory concentration of the sample against microorganisms.

Table 8 shows the result of the test method as described above. The unit is ppm.

[Table 8]

Next, Test Example 2 using a natural bacteriostatic substance will be described. This Test Example 2 is a measurement of the effect of a natural bacteriostatic substance on the growth of Escherichia coli.

The effect of natural bacteriostatic substances on the growth of Escherichia coli was investigated in LB liquid medium over time, and the results are shown in FIG. A typical sigmoid-like growth curve was drawn in the medium inoculated with the inoculum of Escherichia coli in the medium without addition of the natural bacteriostat, but the growth of Escherichia coli was significantly suppressed in the presence of the natural bacteriostat. When the concentration of the natural bacteriostatic substance was 0.0025% or more, no growth of the inoculated Escherichia coli was observed even after culturing at 37 degrees Celsius for 2 days.

The effects of natural bacteriostatic substances can be understood by the above test examples. In addition, the concentration for various microorganisms can be known.

[0044]

As described in detail above, the present invention has the following effects. According to the first aspect of the present invention, the citrus paradecy seeds can be industrially obtained in a large amount and can be obtained at a low cost, so that the natural antibacterial substance to be contained in the spray container can be produced in a large amount at a low cost. In particular, since this citrus paradesi is food itself, it is safe when sprayed in the form of mist in the interior, car, food processing plant, kitchen, hospital, etc. There is no limit to the concentration used. That is, it has flexibility in use. In other words, it has the advantage of having an extremely high bacteriostatic action.

According to the inventions of claims 2 and 3, citrus paradisi is a food that is easily available, and has the advantage that it is easily available in large quantities at low cost.

According to the invention of claim 4, there is an advantage that the natural antibacterial substance can be easily sprayed in the form of mist.

According to the fifth aspect of the present invention, there is an advantage that it is easy to spray a wide range on the bacteriostatic target.

According to the invention of claim 6, it is very easy to take out the seeds of citrus paradecy, to crush the seeds, and to extract and extract the extract from the crushed seeds. It has an advantage that it can be mass-produced and is highly safe in manufacturing.

[Brief description of drawings]

FIG. 1 is a graph showing a growth inhibitory effect of natural bacteriostatic substances on Escherichia coli.

Claims (6)

[Claims] 1. A spray comprising a bacteriostatic substance for sterilizing bacteria present in the bacteriostatic target by spraying the bacteriostatic target in a mist form, and a container containing the bacteriostatic substance.
A spray comprising a natural bacteriostatic substance, characterized in that the bacteriostatic substance is an aqueous solution of an extract extracted from crushed citrus paradecy seeds. 2. A spray comprising a natural bacteriostatic substance according to claim 1, characterized in that the citrus paradesy is grapefruit. 3. A spray comprising a natural bacteriostatic substance according to claim 1, characterized in that said citrus paradecy is a pomelo species. 4. The spray is equipped with a pressurizing pump in order to spray an aqueous solution of an extract extracted from crushed citrus paradecy seeds to the outside from the inside of the container in a mist form. A spray comprising the natural bacteriostatic substance according to claim 1. 5. A spray comprising a natural bacteriostatic substance according to claim 4, wherein the spray comprises an aqueous solution of an extract extracted from crushed citrus paradecy seeds and a filling gas. 6. A method for producing a spray comprising a bacteriostatic substance for sterilizing bacteria present in the bacteriostatic target by spraying the bacteriostatic target in the form of mist, and a container containing the bacteriostatic substance. The bacteriostatic substance is a natural bacteriostatic substance characterized by being obtained by extracting seeds from citrus paradecy, crushing the extracted seeds, extracting the liquid from the crushed seeds, and diluting it with water. A method of making a spray consisting of.