JPH02286609A - Antimicrobial composition - Google Patents

Abstract

PURPOSE:To obtain an antimicrobial composition obtained from a natural material, having no effect on flavor or taste of a food and having strong antimicrobial properties and useful as a preservative for food or the like by extracting grape with water or mixed solvent of water and organic solvent. CONSTITUTION:The novel antimicrobial composition obtained by extracting a grape with water or mixed solvent of water and organic solvent (e.g. methanol, ethanol or acetone). The grape and extracting solvent are preferably used at a ratio of extracting solvent of about 3-20 pts.wt. based on 1 pt.wt. grape. Although the grape may be directly used, when dried grape, pericarp left after removing the juice from the grape, seed or peduncle is used it is advantageous from the viewpoint of handling properties, yield of extract, etc. The antimicrobial composition can be sufficiently utilized for food as well as cosmetic, animal feed, industrial good.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a novel antibacterial composition obtained by extraction from grapes.

[Conventional technology]

Traditionally, sorbic acid,
Chemically synthesized products such as benzoic acid and propionic acid are permitted as food additives, but the applicable foods are limited, and because they are chemically synthesized products, their use may give a negative image to the food product. For these reasons, natural, low-toxicity preservatives are required.

The existence of antibacterial substances in natural products has been known for a long time, and their utility as food preservatives has been studied.

For example, many spices are known to contain strong antibacterial substances.

However, spices often have a negative effect on the flavor and taste of the food itself due to their unique flavor and taste, and are only used as preservatives for a limited number of foods.

[Problem to be solved by the invention]

An object of the present invention is to provide a substance obtained from natural products that does not adversely affect the flavor or taste of foods and has strong antibacterial properties.

[Means to solve the problem]

The present invention is an antibacterial composition obtained by extracting grapes with water or a mixed solvent of water and an organic solvent.

Although the grapes used in the present invention may be fresh, it is preferable to use dried raisins for storage, or the pericarp, seeds, and stems that remain after removing the juice, for ease of handling and yield of extract. It is convenient from

For extraction, water or a mixed solvent of water and an organic solvent such as methanol, ethanol, n-propanol, isopropanol, acetone, etc. can be used. The ratio of the organic solvent is preferably in the range of 0 to 80% (by volume). If the proportion of the organic solvent is higher than this, it becomes difficult to extract the water-soluble antibacterial composition. The ratio of grapes to extraction solvent may be within a range that does not interfere with the extraction operation. Preferably, 3 to 20 parts by weight of the extraction solvent is used per 1 part by weight of grapes.

A centrifugal separation method or a filtration method is used to obtain the extract, and a filter aid such as diatomaceous earth or bentonite can be used during the filtration operation. Further, the extract may be colored depending on the grape raw material used, and in this case, it is preferable to treat the extract with activated carbon or other suitable dye adsorbent to decolorize it. In this respect, the antibacterial composition of the invention is composed of components different from the grape pigment, which is already a natural additive.

The extracted antibacterial composition may be concentrated and dried as it is, or may be dried and powdered after dissolving ills, water-soluble dextrins, etc. in order to improve dissolution and dispersibility in foods.

The antibacterial composition of the present invention has an R7 value of 0.84.0.58.0.51 in a chromatogram obtained by thin layer chromatography using benzene:ethanol mixed solvent (5:1) as a developing solvent and silica gel as a carrier. It consists of a compound of Therefore, the antibacterial composition of the present invention is not a single composition but is composed of a plurality of components. However, each component compound having these R2 values is also included in the present invention.

The antibacterial composition of the present invention can be used as a preservative for all kinds of foods, including various fish paste products, side dishes, pickles, and confectionery, and there are no restrictions at all.

The antibacterial composition of the present invention can be fully utilized not only for foods but also for non-food items such as cosmetics, animal feed, industrial chemicals, etc.

〔Example〕

EXAMPLES Hereinafter, the present invention will be explained in more detail with reference to Examples.

In the examples, percentages are based on weight unless otherwise specified.

Example 1 The skin, stem (after being sun-dried for about 2 months and stored at room temperature) and seeds (after being air-dried at night and stored at -15°C) of Koshu grapes were each ground. 10 grams of 50 volume % acetone per 1 kg of each crushed tissue! , was added and extracted by stirring for 1 hour.

Next, this was suction-filtered, 2% Celite was added to the filtrate, and the mixture was further suction-filtered to obtain a clear extract. This extract was concentrated under reduced pressure and then lyophilized to obtain a light brown powder. The yield of powder obtained from each tissue was 6.1 g from the pericarp, 14.7 g from the stem, and 29.6 g from the seed.

An aqueous solution of this powder was added to a liquid medium [meat extract 0. 3%, peptone 0.5%, magnesium sulfate (MgS04 ・7
H20) 0.1%, 0.0005% to 0 at pH 7,03
．． 05%, and the effect on the growth of Bacillus subtilis was investigated at 35°C.

The results are shown in Figures 1.2 and 3.

Furthermore, Fig. 1 shows the effect of pericarp extract on the growth of Bacillus subtilis, Fig. 2 shows that of rachis extract, and Fig. 3 shows that of seed extract.

As shown in Figure 1, the pericarp extract is Antibacterial properties are observed when 0.005% is added.

Furthermore, as is clear from FIGS. 1 and 2, the addition of 0.101% of the rachis extract and the addition of 0.005% of the pericarp extract have approximately the same antibacterial activity.

Furthermore, as shown in FIGS. 1 and 3, the addition of 0.01% seed extract and the addition of 0.005% pericarp extract have comparable antibacterial activity.

Next, the Koshu grape skin extract was fractionated by thin layer chromatography on a silica gel carrier.

Here, the thin layer used was 20×20 cm with a thickness of 0.4 mm. An ethanol solution containing 30 μm of the extract was spotted in a strip, and developed at 25° C. for 30 minutes using benzene/ethanol (5:1) as a solvent. After air drying, bands were detected under UV light (365 nm).

The results are shown in FIG. That is, R is 0.84.
Three bands of 0.58 and 0.51 are detected.

Next, as shown in Figure 4, it is divided into five fractions, each fraction is scraped off, the extract is eluted with ethanol, concentrated to about 1/10 in an evaporator, and then passed through a membrane filter (
0.45 μm) and used as a sample.

The yield for each portion was 1:16 mg (8,9%) of fraction per 6 thin layers, 180 μ of extract, 2:44 μ (24,
5%), fraction 3: 47 mg (26,0%), fraction 4: 5
0 mg (27,8%), fraction 5: 36 mg (20,1%
)Met.

Both amounts were added to the culture medium at a concentration of 0.01%, and Bacillus subtilis
) was used as a test bacterium to examine its antibacterial activity.

The results are shown in FIG.

As shown in Figure 5, fraction 2 has the highest activity (and fraction 3 follows. Fraction 2 and fraction 3 together account for about 50% of the total activity.
%.

Example 2 Riesling, Saint-Emilion, Chardonnay, Koshu, Merlot, Muscat Heli-A, Cabernet Sauvignon,
The antibacterial properties of an aqueous ethanol extract were investigated using Cabernet Franc grapes.

That is, the skins, stems, and seeds that are obtained as residual aroma when tasting various wines are washed with water, air-dried in the shade, and ground in a coffee mill. 40 to 50% ethanol is added to the powder, and the mixture is shaken at room temperature for 30 minutes. This was filtered through Toyo No. 5c (manufactured by Toyo Roshi), and the filtrate was concentrated under reduced pressure until it became a slag.

This was dried in a high back oven (60°C) to obtain a powder.

Place 1 inch of the obtained dry powder into an L-shaped tube and add 9.9% of Nutrient Broth (beef extract 0.3%, peptone 0.5%, magnesium sulfate 0.1%, pH 5.5). After adding 5 mff1, sealing with a silicone stopper and autoclaving, the bacteria [Bacillus subtili
s) 0.5 ml of IAM 1069) suspension (1 drop of preculture solution (pH 6, 8, 30°C, 110-12 hours) in 10 mR sterile water) was inoculated using a dry sterilized volumetric pipette. This time was defined as 0 hours. , shaking culture was performed at 30°C.

After 6 hours, 1 mfl was sampled and diluted with 9 ml of sterile water to give a viable count of 500 cells/level. This l
ml to a sterile petri dish, and add 0.3% beef extract, 0.5% peptone, 0.0ml magnesium sulfate to a sterile agar medium.
1% agar, 1.3% agar, pH 6,8) was added, and after culturing at 37°C for 148 hours, the number of viable bacteria was counted.

The results are shown in Table 1.

Powder extracted from the pericarp, stem, and seeds of each grape variety 0.2
When 0.02% was added, clear antibacterial activity was observed in all results, but when 0.02% was added, there were considerable differences depending on the grape variety and tissue.

Table 1-1 Table 1-2 Table 1-3 Example 3 Mixture 1 of Koshu grape skin, stem, and dried seeds
70% methanol 10I per kg! , and extracted using a homogenizer at 2000 rpm for 10 minutes. The suspension was filtered under pressure with nitrogen. Activated carbon was added to the filtrate to give a concentration of 2%, and after stirring for 15 minutes, the mixture was filtered under pressure with nitrogen.

The filtrate was concentrated under reduced pressure at 50°C, and the refractive index of the concentrated liquid was measured while removing methanol, and the filtrate was concentrated until the refractive index became 60 to 70%. Two times the amount of pure methanol was added to the concentrate, and the resulting precipitate was filtered off and concentrated to dryness under reduced pressure to obtain 107 g of powder. 0.05-0.5% of this powder
Trypto soy agar medium containing (tryptone 1.7%, peptone 0.3%, glucose 0.28%, potassium dihydrogen phosphate 0.25%, sodium chloride 0.5%, agar 1.3
%) 10mffi was dispensed into a sterile petri dish.

After solidification, 1 platinum loop of test bacteria suspension (culture solution obtained by taking 1 platinum loop from slant culture, inoculating it into 10 mk Tristosoy liquid medium (pH 7,0), and culturing with shaking at 37°C for 6 hours) was smeared. The cells were cultured at 37°C for 172 hours, and the growth of the bacteria was determined by the presence or absence of colonies. The results are shown in Table 2.

As shown in Table 2, the antibacterial composition of the present invention suppresses the growth of Durum-positive bacteria and Durum-negative bacteria at an addition amount of 0.3%.

Table 2; No growth +; Growth, Bacillus 5ubtilis Staphylococcus aureogeus S
almonella typhimurium Ente
robactor aerogenes Escheri
chia coli Pseudomonas aeruginosa [Effects of the Invention] The antibacterial composition of the present invention does not have any adverse effect on the flavor or taste of food even when added to food, so it satisfies the demands of the food industry and is highly appreciated by the industry. It brings great benefits.

[Brief explanation of drawings]

Figure 1 shows the effect of Koshu grape skin extract on the growth of Bacillus subtilis, Figure 2 shows the effect of Koshu grape pericarp extract on the growth of Bacillus subtilis, and Figure 3 shows the effect of Koshu grape skin extract on the growth of Bacillus subtilis. The effect on growth on the same side is shown. FIG. 4 shows a thin layer chromatogram of Koshu grape skin extract. FIG. 5 shows the antibacterial activity of each fraction obtained by thin layer chromatography. Patent Applicant Asama Kasei Co., Ltd. Agent Patent Attorney Shige Takashi Shirai Agent Patent Attorney Kei Sakaguchi Figure 761

Claims (1)

[Claims] (1) An antibacterial composition obtained by extracting grapes with water or a mixed solvent of water and an organic solvent.