JPH07250658A - Antimicrobial agent - Google Patents

Abstract

PURPOSE:To obtain an antimicrobial agent containing components extracted from the fruit shell of mangosteen with water or a water-soluble solvent as the active components, reduced in PH-dependany, having an antimicrobial effect on wideranging microorganisms and useful for reduction of the viable cell number in a food. CONSTITUTION:This antimicrobial agent contains components obtained by extracting the fruit shell of mangosteen with water or an aqueous solvent (methanol, ethanol, water/methanol mixture liquid, water/ethanol mixture liquid, etc.) as the active components. The active components are added to a food, etc., in an amount of 0.01 to 1.0wt.% on solid matter base.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a natural antibacterial agent derived from mangosteen, which has a growth inhibitory effect on a wide range of microorganisms.

[0002]

2. Description of the Related Art Conventionally, various antibacterial agents have been used to improve the storability of foods. As a synthetic antibacterial agent, the use of sorbate, benzoate, etc. has been approved and is used according to the purpose. But these are p
The antibacterial activity is weaker from neutral to weakly acidic than H = 4 to 5, and the effect is generally greatly influenced by pH. Therefore, combined use with salt, ethanol, organic acid, etc., or multiple antibacterial agents A combination is needed. In recent years, although synthetic antibacterial agents have excellent effects, the safety remains uncertain, and there is a growing need for safer natural antibacterial agents.For example, formulations of lysozyme, protamine, polylysine, spice extracts, etc. have been developed. , Has been commercialized. However, these natural antibacterial agents are generally characterized by a weaker antibacterial effect and a narrower antibacterial spectrum than synthetic antibacterial agents. In addition to this, the sensory aspects such as taste and smell and the cost aspect are inferior to the present level, and it is currently only a supporting role for synthetic antibacterial agents. The effect of natural antibacterial agents is greatly influenced by pH, as is the case with synthetic antibacterial agents.

On the other hand, a water-soluble extract of mangosteen fruit shell is known to have a whitening effect and an anti-inflammatory effect (Japanese Patent Laid-Open No. 4-244004). Furthermore, the organic solvent extract of mangosteen fruit shell contains various xanthones,
It is also known to have anti-inflammatory, anti-immunosuppressive and anti-diarrheal effects (Jones, WD, Albrecht, W.
L., Monro, NLand Stewart, KT: J.Med.Chem.20,594 (19
77) ； Balasubramanian, K. And Rajagopalan, K.: Phytoche
mistry.27 (5): 1552-1554 (1988) and others). However, nothing is known about the use of water or a water-soluble solvent extract of mangosteen fruit husks as an antibacterial agent.

[0004]

The object of the present invention is excellent in safety and stability against pH, can be added to foods, cosmetics, etc., and has a versatile property of inhibiting growth against a wide range of microorganisms. To provide an antibacterial agent.

The present inventors have conducted various studies to solve these problems, and as a result, an antibacterial agent containing an extract obtained by extracting mangosteen fruit shells with water or a water-soluble solvent as an active ingredient has an extremely excellent antibacterial activity. Found to have activity,
The present invention has been completed.

[0006]

That is, the present invention provides an antibacterial agent characterized by containing, as an active ingredient, a component extracted from mangosteen fruit shells with water or a water-soluble solvent, and mangosteen fruit shells containing water or water. The present invention relates to a method of using a component extracted with a water-soluble solvent as an antibacterial agent.

The mangosteen used in the present invention is a mangosteen of the Hypericumaceae family, and its scientific name is Garcinia man.
Gostana L., a plant widely cultivated in Southeast Asia and tropical Asia.

The mangosteen fruit shell used in the present invention is
The fruit shell is obtained by removing the pulp from mangosteen fruit, and the antibacterial agent of the present invention can be obtained by the following method using the fruit shell as a raw material.

First, the obtained mangosteen fruit husk is crushed to an appropriate size. Subsequently, this fruit shell may be directly subjected to the extraction operation using the extraction solvent, but it is preferable to perform the extraction operation after drying.

The extraction solvent used in the present invention may be water or a water-soluble solvent. Examples of the water-soluble solvent include lower alcohols such as methanol, ethanol and propanol, and acetone. Further, a mixed solvent thereof can be used. The extraction solvent can be appropriately selected and used, but due to cost and ease of handling,
Water, methanol, ethanol, a water / methanol mixed solution, and a water / ethanol mixed solution are preferable. The mixing ratio of the mixed liquid is arbitrary, but in view of extraction efficiency, water: methanol or water: ethanol is 40:60 to 60:40 in volume ratio.
Is preferred.

Extraction can be carried out by immersing the mangosteen fruit shell in such an extraction solvent. During extraction, it is preferable to stir the solvent. The extraction method may be either a heat extraction method or a room temperature extraction method, and the extraction temperature is not particularly limited as long as it does not boil the extraction solvent, but if it is too low, it takes an extraction time, and therefore it is preferably 3
The temperature is 0 to 70 ° C, particularly preferably 40 to 60 ° C.

The amount of mangosteen fruit husks added to the extraction solvent is not particularly limited and may vary depending on the number of extractions, the extraction temperature, the extraction time, etc., but it is preferable to use 2 to 10% by weight of the husks relative to the solvent. It is also preferable from the viewpoint of the surface of distilling off the solvent after extraction.

The number of times of extraction varies depending on the extraction time, the extraction temperature, etc., but is usually 1 to 3 times. The extraction time depends on the extraction temperature, etc.
1 to 5 hours are preferable, and 2 to 3 hours are particularly preferable for each extraction. In the case of extraction at room temperature, it is 10 to 24 hours, preferably 15 to 20 hours.

Next, the extraction solvent, fruit shells,
The residue is filtered from the mixture of extracts, and the filtrate is concentrated or diluted as needed to obtain a liquid, paste-like, or solid antibacterial water-soluble extract of mangosteen fruit husks. The obtained extract can be used as it is or after purification as an antibacterial agent.

The antibacterial agent of the present invention may contain the above water-soluble extract as an active ingredient, and the extract alone can be used as an antibacterial agent. It can be mixed with the additives used in (1) and formulated by a known method to be used. Further, food additives and the like can be added to the antibacterial agent.

The antibacterial agent of the present invention contains various foods, health foods,
It can be added to drugs, quasi-drugs, cosmetics, livestock feed, pet food, etc. that require an antibacterial agent, and it is preferable to add it to various foods because it is a natural product and excellent in safety. By adding to these, spoilage due to fungi can be prevented.

Examples of foods to which the antibacterial agent of the present invention can be added include ordinary foods in general, for example, fish paste products such as kamaboko, bamboo rings, date rolls, tsumi, hanpen, and Satsuma fried, ham, sausage, bacon, beef jerky. , Whale meat and meat products such as hamburger steak and meatballs,
Delicacies such as squid, octopus smoked, grilled fish, dried fish, raw sea urchin, steamed sea urchin, seafood, kelp Tsukudani, seaweed Tsukudani, shellfish Tsukudani, Takuan pickle, koji pickle, soy sauce pickled, salted, miso pickled, Pickles such as pickled vegetables, dairy products such as cheese, yogurt, butter, custard cream and whipped cream, wine, sweet sake, carbonated drinks, non-carbonated drinks, fermented milk and other drinks, dressings, fruit sauces, ketchups, sauces such as sauces Kind,
Examples include seasonings such as soy sauce and miso, high-sugar-concentration foods such as jam and jelly, as well as raw noodles, breads, confectionery, and prepared foods.

Regarding the method of adding the antibacterial agent of the present invention, even if it is added in advance to the material of the additive such as food,
It may be added during the production of the additive such as food, and may be appropriately selected in consideration of workability.

The antibacterial agent of the present invention is used in a solid content of 0.01 to 1.0% by weight, preferably 0.05 to 0.5% by weight, based on the whole additive such as food. Appropriate. If the amount is less than 0.01% by weight, no sufficient effect can be expected, and if the amount exceeds 1.0% by weight, the effect is not enhanced depending on the bacterial species and the efficiency is poor.

The antibacterial agent of the present invention has a wide range of effects on various fungi, and examples of bacteria include Escherichia c
oli), Enterobacter cloacae
acae), Klebsiella pne
umoniae), Pseudomonas aeruginosa, and acetic acid bacteria (A
Gram-negative bacteria such as cetobacter aceti), Staphylococcus aureus, and Bacillus subtilis
tilis), Clostridium botulinus and other Gram-positive bacteria, and as a fungus, Aspergill
us niger), Rhizopus and other species
Genus), molds such as Penicillium (genus Penicillium) such as blue mold, yeasts such as Saccharomyces cerevisiae. Since the antibacterial agent of the present application is derived from a natural product and has high safety, it is particularly useful as an antibacterial agent for food additives.

[0021]

EXAMPLES Examples will be shown below, but the present invention is not limited to these examples.

Example 1 Dried mangosteen fruit shell 2
0 g was crushed, heated and extracted with 500 ml of water at 50 ° C. for 3 hours, filtered, and concentrated under reduced pressure to obtain 1.68 g of a brown solid substance.

Example 2 Dried mangosteen fruit shell 2
Crush 0g and mix with water / ethanol (volume ratio 1: 1) 5
The mixture was heated and extracted with 00 ml at 50 ° C. for 3 hours, filtered, and concentrated under reduced pressure to give a brown solid 3.16.
g was obtained.

Example 3 Dried mangosteen fruit shell 2
0 g was crushed, heat-extracted with 500 ml of ethanol at 50 ° C. for 3 hours, filtered, and concentrated under reduced pressure to obtain 2.04 g of a brown solid substance.

Example 4 Dried mangosteen fruit shell 2
0g was crushed and mixed with water / methanol (volume ratio 1: 1) 5
The mixture was heated and extracted with 00 ml at 50 ° C. for 3 hours, filtered, and concentrated under reduced pressure to give a brown solid 2.88.
g was obtained.

(Example 5) Dried mangosteen fruit shell 2
0 g was crushed, extracted with 500 ml of methanol at room temperature for 18 hours, filtered, and concentrated under reduced pressure to obtain 1.40 g of a dark brown solid substance.

Example 6 Dried mangosteen fruit shell 2
Crush 0g and mix with water / acetone mixture (volume ratio 1: 1) 50
Extraction was performed with 0 ml at 50 ° C. for 3 hours, followed by filtration and concentration under reduced pressure to give a brown solid 0.88.
g was obtained.

Test Example 1 Using the extracts obtained in each of the above Examples, their antibacterial activities were evaluated by the following experiments. The medium used was tryptosoy broth medium (Eiken Chemical Co., Ltd.) to which agar (manufactured by Wako Pure Chemical Industries, Ltd.) was added, and each extract had a final concentration of 0.01 to 1.0% by weight. And sterilized. In addition, in order to confirm the effect of pH, the pH of the medium before sterilization was adjusted so that the final pH after sterilization was 6 or 7. The prepared medium is dispensed into a petri dish and solidified, and then a diluted solution of the suspension of the test bacterium that has been liquid-cultured in advance (about 200 to 500 cells / dish: adjusted to 0.1 ml) is agar medium. Inoculated on. Bacteria used were 3 types of bacteria and 2 types of fungi shown in Table 1. Bacteria were cultivated at 37 ° C for 48 hours and at 30 ° C for 72 hours, the extract concentration required to inhibit the growth of the bacteria was determined. Was measured. The results are shown in Table 1. The pH in the table indicates the pH of the medium after addition of the antibacterial agent obtained in each Example and sterilization, and the numerical values in the column are expressed as the minimum inhibitory concentration (% by weight).

[0029]

[Table 1] Test strain Bacillus subtilis IFO-6633 Escherichia coli IFO-3543 Staphylococcus aureus IFO-12732 Saccharomyces cerevisiae IFO-0213 Aspergillus niger ATCC-6275

From the above results, the effect of the antibacterial agent of the present invention is as follows.
It was confirmed that it is stable to pH and has an excellent growth-inhibiting effect even on a wide range of microorganisms. Thus, the antibacterial agent of the present invention is effective against various bacteria and fungi related to spoilage, including Staphylococcus aureus, which is one of food poisoning bacteria.

(Comparative Example 1) By the same method as in Test Example 1, the concentration of potassium sorbate required for inhibiting the growth of bacteria was measured. The results are shown in Table 2. The pH in the table is the medium pH after sterilization by adding the antibacterial agent of each test example.
The numerical value in the column is represented by the minimum inhibitory concentration (% by weight).

Comparative Example 2 By the same method as in Test Example 1, 40% by weight of ethanol, 10% by weight of acetic acid and 50% by weight of water were used.
Was mixed to prepare a formulation, and the concentration necessary for inhibiting the growth of the fungus was measured. The results are shown in Table 2. Since the formulation of this comparative example had a possibility that the composition changed when autoclaved along with the medium, only the comparative example after autoclaving the medium after autoclaving to a predetermined concentration before dispensing and solidifying the medium A plate for evaluation was prepared by adding the formulation and gently mixing with the medium.

Comparative Example 3 In the same manner as in Test Example 1, the concentration of sodium benzoate required to inhibit the growth of bacteria was measured. The results are shown in Table 2.

[0034]

[Table 2] Test strain Bacillus subtilis IFO-6633 Escherichia coli IFO-3543 Staphylococcus aureus IFO-12732 Saccharomyces cerevisiae IFO-0213 Aspergillus niger ATCC-6275

From the above results, it was confirmed that the effect of the synthetic antibacterial agent depends on pH as compared with the antibacterial agent of the present invention. Further, its antibacterial activity was often significantly lower than that of the antibacterial agent of the present invention.

(Test Example 2) 0.5% of the mangosteen husk extract of Example 1 was added to the custard cream produced according to a conventional method to examine the antibacterial effect. For comparison, a non-addition group and a 0.5% potassium sorbate addition group were used.
The mixing ratio of custard cream is shown in Table 3. These 3
The seed custard cream was stored at 37 ° C., and the number of general viable bacteria in the custard cream was measured every day. In measuring the number of general viable bacteria, a standard agar medium (manufactured by Nissui Pharmaceutical Co., Ltd.) is used as the medium, and after sterilization, 50 is used so as not to solidify.
The sample was incubated at about ℃, mixed with physiological saline and incubated at 37 ℃ for 48 hours, and the number of colonies that appeared was counted to calculate the number of general viable bacteria in 1 g of custard cream. The results are shown in Table 4.

[0037]

[Table 3]

[0038]

[Table 4]

[0039]

INDUSTRIAL APPLICABILITY The antibacterial agent of the present invention is highly stable against pH and has an excellent growth inhibitory action against a wide range of microorganisms. In addition, it is derived from natural products and is highly safe. By adding it to foods, etc., it suppresses the number of viable bacteria in foods and is used as a powerful antibacterial agent.

Claims (4)

[Claims] 1. An antibacterial agent, which comprises as an active ingredient a component extracted from mangosteen fruit shell with water or a water-soluble solvent. 2. The antibacterial agent according to claim 1, wherein the water-soluble solvent is a lower alcohol. 3. The antibacterial agent according to claim 1, wherein the water-soluble solvent is water-containing methanol or water-containing ethanol. 4. A method of using a component extracted from mangosteen fruit shell with water or a water-soluble solvent as an antibacterial agent.