JPH0827005A - Antimicrobial agent - Google Patents

Abstract

PURPOSE:To obtain an antimicrobial agent, using beta-D-glucopyranoxy- phloroglucinol which is a natural organic compound, capable of manifesting antimicrobial properties against microorganisms such as dental carious bacteria, periodontotic bacteria or molds parasitic on plants, derived from natural products and having high safety. CONSTITUTION:This antimicrobial agent contains beta-D-glucopyranoxy- phloroglucinol of the formula as an active ingredient. The antimicrobial agent is useful as a durability improving agent for a food and drink and/or an antimicrobial agent and effective against dental carious bacteria and periodontotic pathogenic bacteria and further acts as an antifungal agent. The antimicrobial agent is useful as an antifungal agent in storing and transporting especially vegetables and fruits.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an antibacterial agent, and more particularly to a highly safe antibacterial agent containing a natural organic compound as an active ingredient.

[0002]

2. Description of the Related Art Conventionally, it is derived from natural products, especially citrus peels, is easily soluble in water, has no taste and odor, and is stable to heat.
As for the phloroglucinol-based antibacterial substance having excellent antibacterial activity and high safety, no examples have been known in which the structure was successfully isolated by isolating and purifying it.

On the other hand, β-D-glucopyranoxy phloroglucinol (hereinafter sometimes referred to as LE-V) is
For having excellent antibacterial properties against microorganisms (eg, caries, periodontal bacteria and fungi parasitic on plants),
Neither patents nor other research reports have been made.

[0004]

At present, a chemically synthesized product is unavoidably used as an antibacterial agent, but there is a problem in terms of safety for human body and environment. Therefore, in the industry, development of an excellent antibacterial agent which is highly safe, easy to handle, and has a wide application range is desired.

[0005]

The present invention has been made in order to meet the needs of the above-mentioned industry, and attention was paid to natural products in terms of safety.

[0006] Therefore, the inventors of the present invention have conducted extensive studies in search of an antibacterial substance derived from a natural product, and have confirmed that the peel of citrus fruits such as lemon, grapefruit, valencia orange and navel orange contains the antibacterial substance. Of. And, not only succeeded in separating and purifying the antibacterial substance, but also succeeding in determining the structure of the purified substance (LE-V), and further confirming the excellent antibacterial property of this purified substance, The invention was completed.

That is, the present invention relates to an antibacterial agent containing LE-V as an active ingredient. The LE-V substance is a natural substance derived from citrus peel and is a phloroglucinol derivative, and as a result of the structure determination, β-D-glucopyranoxy phloroglucinol (β-D-glucopyranoxy ph
loroglucinol) was identified.

LE-V is lemon, grapefruit,
It is present in squeezed lees of citrus fruits such as Valencia orange and navel orange, as well as in the peel and juice, and can be extracted and purified from these. As the extraction solvent, a solvent such as water, hot water, alkaline water or acidic water, or an organic solvent such as methanol, ethanol or acetone can be used for extraction. Even in the crudely purified state, the antimicrobial effect is exhibited if the LE-V component is in a sufficient amount. The LE-V substance is not only highly safe and has an excellent antibacterial action, but also has a broad antibacterial spectrum, and thus can be widely used as various antibacterial agents. Moreover, it is easily soluble in water, tasteless, odorless, and stable to heat, so it is extremely easy to use and can be used in a very wide range of applications.

For example, the LE-V substance can be widely used for the following purposes. Adds directly or indirectly to food and drink to improve shelf life or use as an antibacterial agent;
Antibacterial agents against periodontal pathogens (for example, drugs applied directly to the affected area, mouthwash, mouthwash, and toothpaste); antifungal agents; antifungal agents during storage and transportation of fruits and vegetables (For example, as a coating agent, a wax agent, and a substitute for a post-harvest pesticide).
Examples of the present invention will be described below.

[0010]

Example 1 The peel of lemon squeezed lees crushed with a mixer was allowed to stand in ethanol at 37 ° C. for 3 days for extraction. This was distilled under reduced pressure with an evaporator to remove ethanol. This was dissolved in water, and liquid-liquid solvent fractionation was performed using a separating funnel.

In the liquid-liquid solvent fraction, hexane, chloroform, ethyl acetate, water-saturated butanol and water were used as a solvent, and fractionation was carried out sequentially to obtain respective soluble fractions. Of these fractions, the water-saturated butanol fraction was distilled under reduced pressure to dryness and then dissolved in water. This was further fractionated by preparative HPLC to obtain a fraction showing activity.
By repeating preparative HPLC for this fraction,
Purified material (LE-V) was obtained. This was freeze-dried to prepare a powder product.

Regarding the obtained LE-V substance, ¹ H N
MR analysis, ¹³ C NMR analysis and other analyzes were performed to obtain the physicochemical properties shown in Table 1 below.

[0013]

[Table 1]

From these results, the structure of LE-V was determined, and LE-V was β-D-glucopyranoxy phloroglucinol.
Was identified. Its chemical structural formula is as shown in Chemical Formula 2 below.

[0015]

Embedded image

[0016]

Example 2 The antibacterial activity of the LE-V substance against caries and periodontal disease bacteria was measured and confirmed as follows.

Regarding tooth decay, St known as the causative fungus
reptococcus mutans ATCC 7270 was used. For periodontal disease, we used bacteria that cause gingivitis and alveolar pyorrhea. That is,
Prevotella intermedia and A are the causative bacteria of gingivitis.
Cinomyces viscosus NIAH 1010 was used, and Porphyromonas gingivalis 381 was used as the causative bacterium of alveolar pyorrhea. Brain heart infusion (BHI) medium was used as the growth medium for S. mutans and A. viscosus, and gum (GAM) medium was used as the growth medium for P. intermedia and P. gingivalis.

To measure the growth inhibitory activity of the LE-V substance against these fungi, 10 ⁵ cells / ml of the bacteria were inoculated into ⁵ ml of broth medium added at various concentrations, and the mixture was incubated at 37 ° C. for 2 hours.
It was cultured for one day. However, since P. gingivalls is an obligate anaerobic bacterium, the anaerobic jar (BB
L). Then, the culture solution was added to an agar medium at 0.
After adding 1 ml and culturing this at 37 ° C. for 2 days, the presence or absence of colonies was observed.

Antibacterial property was judged to be sterilized when there was no colony. Then, the correlation between the sample concentration and the antibacterial effect was examined, and the strength of the activity was indicated by the minimum inhibitory concentration (MIC) for sterilizing the same bacteria at the minimum concentration. The obtained results are shown in Table 2 below, and LE-V was found to have excellent growth inhibitory effect and sterilizing effect against dental caries and periodontal disease bacteria.

[0020]

[Table 2]

[0021]

Example 3 The growth inhibitory activity of the LE-V substance against the fungi generated during fruit storage was measured and confirmed as follows.

Test moulds were isolated from moldy stored lemon fruits. And Penicillium digi with green mold
Spores mixed with tatum and blue mold Penicillium italicum were obtained. The growth medium is potato dextrose agar (P
DA) medium was used.

The growth inhibitory activity of the LE-V substance against these molds was measured by 5 ml of an agar medium supplemented with LE-V.
Was made into a petri dish (4.5 cm in diameter), and a hole having a diameter of 2 mm was opened at the center of the petri dish. There 10 ⁵ spores / ml
Mold fungus spores of 5 μl each at 25 ° C
Cultured for 7 days. Measure the circle diameter of the mycelium of the grown mold,
The degree of proliferation was observed. The results are shown in Table 3 below.
It was confirmed that -V has an excellent mold growth inhibitory effect.

[0024]

[Table 3]

[0025]

Example 4 The antibacterial property of LE-V against microorganisms (bacteria, yeast) was measured and confirmed as follows.

As bacteria, Lactobacillus casei, Strept
ococcus lactis is Escherichia coli IFO in GAM medium
3301, Salmonella typhimurium IIP 1000, Bacillus s
ubtilis AJ 1235 was cultured in NB medium. As yeast, S
accharomyces cerevisiae IFO 1346, Hansenula anomal
a IAM 12914 was cultured in YM medium. LE-V for each medium
Was added by changing the concentration, and autoclaved (121
℃, 15 minutes), inoculate each bacterial suspension, bacteria at 37 ℃,
The yeast was cultured at 25 ° C. and the sterilization concentration of LE-V was examined.
The results obtained are shown in Table 4 below.

[0027]

[Table 4]

[0028]

Example 5 Solution A (LE-V 50 g, deionized water 5 k
g), and solution B (sucrose 50 g, citric acid 8 g, sodium citrate 8 g, vitamin C 1 g, flavor 10 g, 1
/ 5 concentrated lemon juice 100 g and water 5 kg) were prepared.

Solution A and solution B were separately held in a plate sterilizer at 90 to 95 ° C for 3 minutes for sterilization, then cooled to 5 to 10 ° C and stored in a tank. Then, aseptically fill the paper container with the mixed solution thus obtained,
A long-term storable lemon juice drink for preventing tooth decay was manufactured.

[0030]

Example 6 A toothpaste for preventing dental caries was produced with the following formulation.

Formulation (%) Calcium hydrogen phosphate dihydrate 45.0 Glycerin 10.0 Sorbitol 25.0 LE-V 2.0 CMC-Na 0.5 Carrageenan 0.5 Saccharin sodium 0.2 Lauryl sulfate Sodium 1.2 Vitamin E acetate 0.1 Peppermint flavor 1.0 Water balance

[0032]

Example 7 A LE-V blended preparation was prepared using LE-V 25.0%, glycerin fatty acid ester 8.0%, sodium acetate 4.5%, and ethanol 62.5%.

This compounded formulation and a commercially available wax formulation (manufactured by Koto) were mixed at a ratio of 2: 3 to prepare an LE-V wax formulation. This formulation is extremely useful for storing citrus fruits such as lemons, oranges, and grapefruits, as well as various fruits and fruits and vegetables, and has a long shelf life as a substitute for post-harvest pesticides that are mainly chemical synthetic products that have been conventionally used. It is also extremely superior in terms of safety.

[0034]

EFFECTS OF THE INVENTION Since the antibacterial agent according to the present invention is derived from a natural product, it has not only excellent antibacterial properties but also extremely high safety, and it is very useful for preventing mildew of fruits and vegetables and antibacterial of foods and drinks. It is valid. Further, since it has a high activity of inhibiting the growth of cavities and periodontal bacteria, it can be appropriately prepared in, for example, toothpaste, mouthwash, and various foods and drinks.

Further, since the structure of the active ingredient of the antibacterial agent was determined by the present invention, it is expected that a novel antibacterial agent will be created by producing a derivative of this compound.

Claims (5)

[Claims] 1. A β-D-glucopyranox phloroglucinol (β-D-glucopyranox) represented by the following chemical formula 1.
yphloroglucinol) as an active ingredient. Embedded image 2. An antibacterial agent and / or a shelf life improver for food and drink.
Or it is an antibacterial agent, The antibacterial agent of Claim 1 characterized by the above-mentioned. 3. The antibacterial agent according to claim 1, wherein the antibacterial agent is an antibacterial agent against caries and periodontal pathogens. 4. The antibacterial agent according to claim 1, wherein the antibacterial agent is an antifungal agent. 5. The antibacterial agent according to claim 1, wherein the antibacterial agent is an antifungal agent during storage and transportation of fruits and vegetables.