KR100799270B1 - Natural anti-microorganism composition containing extract from zanthoxylum schinifolium as active ingredient - Google Patents

Abstract

 The present invention relates to a natural antimicrobial composition comprising an herb extract as an active ingredient. In particular, the present invention has a strong bactericidal effect against food poisoning bacteria, harmless to the human body, and provides a natural antibacterial composition containing the same and excellent thermal stability.

Japanese pepper, food poisoning

Description

NATURAL ANTI-MICROORGANISM COMPOSITION CONTAINING EXTRACT FROM ZANTHOXYLUM SCHINIFOLIUM AS ACTIVE INGREDIENT}

1 is an observation of the antimicrobial activity of the extract of the Japanese pepper with growth inhibition ring,

Figure 2 shows the anti-Vibrio activity when the methanol extract of vinegar except seeds was heat-treated at 60 ℃ (a) or 90 ℃ (b), respectively,

Figure 3 confirms the growth inhibition of Vibrio bacteria of methanol extract mixture of vinegar except 70% ethanol and seeds,

Figure 4 is a vibrio inoculum (A) inoculated in LB medium and vibrio inoculum (B) inoculated in LB medium to which methanol extract was added, except for 10% of the seed was incubated for 37 hours, respectively, and compared the turbidity of the culture solution One,

Figure 5 is a graph showing the growth inhibition effect of Vibrio bacteria by concentration of methanol extract of vinegar except seeds,

FIG. 6 shows the growth of Vibrio bacteria in LB medium containing 10% of each of ethanol methanol extract, clean call, and ethanol methanol extract / clean call (10% acid, 90% clean call composition) except seeds. It is a graph.

[TECHNICAL FIELD OF THE INVENTION]

The present invention relates to a natural antimicrobial composition comprising an extract of the herb as an active ingredient, and more particularly, to a natural antibacterial composition and an antibacterial extract that shows excellent antibacterial activity against food poisoning bacteria.

[Private Technology]

Food poisoning is a disease that occurs around the world, most of which is caused by bacteria. As food poisoning acts as a cause of food poisoning is gradually diversified and the incidence of food poisoning is increasing, interest in preventing food poisoning is increasing. In addition, as the pollution rate increases with environmental pollution, the risk of food poisoning accidents is increasing, and rapid industrialization and population increase threaten the balance of aquatic organisms and ecosystems.

In shellfish grown in contaminated water, the detection rate of food poisoning bacteria such as Staphylococcus aureus and Vibrio parahaemolyticus is high. In addition, looking at the food poisoning incidence occurred in 2000 as the statistics of the Food and Drug Administration, Salmonella, Staphylococcus aureus and enteritis vibrio accounted for more than 50% of the causes of food poisoning, it can be seen that urgent precautions for the three strains are urgently required.

division system Salmonella Staphylococcus Enteritis Vibrio Not detected Etc Number of cases (%) 104 (100) 30 (28.8) 9 (8.7) 14 (13.5) 35 (33.6) 15 (15.4) Number of patients (%) 7,269 (100) 2,591 (35.6) 824 (11.3) 235 (3.2) 2,677 (37.0) 942 (13.0)

In order to prevent the incidence of food poisoning, a lot of chemical synthetic products have been used in the past, but the alternative preservatives that are harmless or harmful to the human body have been developed from natural food materials that have been edible for a long time recently due to evasion due to safety problems of consumers. To this end, a lot of research is being actively conducted. In particular, as incomes rise and living standards rise and health concerns rise, changes in consumer preferences for foods that do not contain synthetic preservatives are facilitating research on natural antimicrobial agents that are harmless to humans. Park, Jong-Cheol (1994), Journal of Biotechnology and Bioengineering, 9 (5), 506-511 (1994); Seok-Cheol Choi, Jin-Suk Jung. (1997) Korean Society of Textile Engineers, 34 (6) p393-399 (1997); Park, Chan-Sung (1998) Journal, 5 (1), p89-96 (1998).

Natural materials commercialized or exhibiting antimicrobial properties include proteins such as lysozyme, polylysine, protamine, cornalbumin, and avidin, organic acids such as acetic acid, banjoic acid, allicic acid, and succinic acid, pectin degradation products, browning reactants, and lower grades. Fatty acid esters and spices (Yang Ji-Young, Jong-Hong Han, Hyun Rok Kang, Mi-Kyung Hwang, Jae-Woo Lee (2001) The Korean Society of Food Hygiene and Safety, 16 (1), p37-40). Many spices are known to have antimicrobial activity against many bacteria, the best known of which are garlic and onion.

However, the development of natural plant-derived material having excellent antimicrobial activity against food poisoning bacteria is still insufficient.

In order to solve the problems of the prior art, an object of the present invention is to provide a plant-derived antibacterial substance having a strong antibacterial activity against food poisoning bacteria.

It is another object of the present invention to provide a plant-derived antibacterial substance that is safe for human body and can be consumed as food without objection.

It is another object of the present invention to provide a plant-derived antimicrobial substance that can be easily added to food poisoning-induced foods to prevent the development of food poisoning.

In order to achieve the above object, the present invention provides an antimicrobial composition comprising an herb extract as an active ingredient.

In another aspect, the present invention provides a food comprising 0.5 to 20% by weight of the antimicrobial composition.

Hereinafter, the present invention will be described in detail.

The present inventors can use it as a substitute for the synthetic antimicrobial compound, and while searching for anti-food poisoning substances that can be safely applied to the human body from natural plants, we confirmed that the Japanese vinegar has excellent antimicrobial activity and completed the present invention. It was.

The herb is a plant belonging to the Unhyang family, the scientific name Zanthoxylum schinifolium , also called the branch or the herb. Japanese pepper is widely grown in Northeast Asia such as Korea, Japan and China, and the bark, leaves and fruits of Japanese pepper are used for spices, medicinal and oil-making (Lee Jong-won (1998), Korean Society of Food and Nutrition, 11 (5), p493-498).

The antimicrobial material of the present invention is an extract of the Japanese herb. The vinegar extract may be prepared by a conventional plant extract manufacturing method, specifically, a volatile extract manufacturing method. In the method of preparing volatile extract, vinegar is crushed, concentrated under reduced pressure by adding ethanol, and volatilized vapor is passed through a cooling tube to be collected in a round flask in the liquid form of condensate, and the collected liquid is concentrated to volatile extract. How to prepare.

The part of the vinegar used for the vinegar extract may be the whole or part of the vinegar, and preferably one or more kinds of fruit bark (leaf), leaves, branches, seeds, roots or stems except seeds, the seed extract Compared to other parts, the antibacterial activity of the extract is better.

The extraction solvent may be distilled water or an organic solvent, and the organic solvent may be lower alcohol having 1 to 5 carbon atoms, hexane, ethyl acetate or chloroform. The extraction solvent may be added in a weight ratio of 1: 1 to 1: 20 to the dry weight of the Japanese anchovy, the extraction solvent may be used alone or in the form of a mixture of one or more, and the extraction process using another solvent sequentially Can be repeated.

Extraction conditions are reduced pressure at high temperature, the temperature is 60 to 250 ℃, preferably 90 to 220 ℃, the pressure is up to 100 mmHg, preferably 10 to 50 mmHg. Extraction time can be appropriately adjusted according to the solvent or the use weight of the vinegar, preferably 6 hours to 48 hours, but is not limited thereto.

The vinegar extract of the present invention is preferably an extract obtained by methanol or ethanol extracting at least one selected portion from the group consisting of barberry, skin, leaves, branches, roots and stems except for the seeds of the vinegar.

The present invention provides an antimicrobial composition comprising an herb extract as an active ingredient. The antimicrobial composition of the present invention preferably has antimicrobial activity against food poisoning bacteria. The antimicrobial composition may include the herb extract alone, and may further include other pharmacologically acceptable substances. The pharmacologically acceptable substance may be an existing antimicrobial or bactericide, and may be a diluent or an excipient. Exemplary antimicrobial agents include proteins such as lysozyme, polylysine, protamine, cornalbumin, and avidin, and organic acids such as acetic acid, banjoic acid, allic acid and succinic acid, pectin degradation products, browning reactants, lower fatty acid esters, or spices. .

The antimicrobial composition of the present invention preferably contains 0.5 to 20% by weight of an herb extract, but the content may be appropriately adjusted according to the type of additive or the formulation of the antimicrobial composition. The formulation of the antimicrobial composition can be appropriately adjusted according to the purpose of use, and representative representative formulations applicable are granules, powders, syrups, solutions, aerosols, emulsions, suspensions or spirits.

In addition, the antimicrobial composition of the present invention may further include an alcohol disinfectant such as alcohol, ethanol, methanol, a commercial alcohol disinfectant. Since the extract of the present invention of the present invention has a synergistic effect of remarkably improving the antimicrobial activity when used in combination with the alcohol-based disinfectant, it is possible to maximize the antimicrobial effect by adding an alcohol-based disinfectant to the antimicrobial composition. A preferred ratio having an optimal synergistic effect is that the mixture of the vinegar extract and the alcohol-based disinfectant in a 1:99 to 1: 4 weight ratio, the synergistic effect is little or out of the weight ratio range may be uneconomical compared to the effect. The alcoholic disinfectant is preferably ethanol.

The antimicrobial composition of the present invention is harmless to the human body and can be used as a food preservative, a food anti-corruption agent or a food disinfectant, and it is very excellent in workability by treating the acidic extract as an active ingredient before heat processing because it is stable to heat.

The present invention also provides a food comprising an antimicrobial composition. The food may contain 0.5 to 20% by weight of the antimicrobial composition, the antimicrobial activity of the food may be insufficient when the content of the antimicrobial composition is less than 0.5% by weight, the effect is not increased when used exceeds 20% by weight It can be uneconomical.

The food of the present invention is a conventional food, preferably a spice, seasoning or wasabi. In particular, wasabi is a food used together with ingestion of sashimi, which is the main cause of food poisoning, and can effectively sterilize harmful strains growing on sashimi.

Hereinafter, examples of the present invention will be described. The following examples are only for illustrating the present invention and the present invention is not limited to the following examples.

Example 1 Preparation of Japanese Herb Extract

1-1. Japanese Herb Water Extract Manufacturer                     

The Japanese herb trees purchased at the Seoul Kyungdong Market were classified into seeds and parts except seeds (dry bark, leaves, branches, roots, and stems), dried, and then crushed. Distilled water was added to each crushed product at a 1:10 weight ratio, concentrated under reduced pressure at 100 ° C, and the obtained volatile extract was filtered through a filter paper (Wathman No. 2). The filtrate was filtered under reduced pressure again using a rotary vacuum evaporator and then concentrated to 1/10 of the volume to be used as a test liquid for the water fractionation layer.

1-2. Manufacture of Methanol Extract

The part of the vinegar except for the seed was crushed, ethanol was added at a 1:10 weight ratio, and concentrated under reduced pressure at 60 ° C. The volatilized vapor was passed through a cooling tube in which water flowed at 4 ° C. was collected in a round flask in the form of a condensate, and the collected solution was concentrated 5 times at 60 ° C. to prepare a volatile extract.

Example 2: Antimicrobial Test of Japanese Herb Extract

The antimicrobial test of the extract was conducted using the disk method (Davidson, P. M and Parish, ME (1989) Food technology , 43, p148-155).

The strains were stored in the laboratory: Shigella flexmari , Serratia liquefacieus , Salmonella enteritidis , Salmonella hedelberg , Salmonella typhimurium ( S. typhimurium ) ATCC14028, S. javiana , Salmonella typhimurium ATCC14023, E. coli O157: H7, 3 Bacillus subtilis , Enterobacter calacae , Staphylococcus aureus Reus ( Staphylococcus aureus ) and Vibrio parahaemolytics were used.

Each of the strains was inoculated in 10 mL LB medium, incubated at 37 ° C. for 16 hours, and 0.1 mL was taken and evenly spread on LB solid medium. 70 μl of the water extract or the methanol extract of the vinegar was dropped on a sterile 10 mm paper disc, dried, and incubated at 37 ° C. for 24 hours to measure the diameter of the clear zone around the disc. At this time, as a control, distilled water and methanol 70 ul were added to the paper disk in the same manner and measured. Table 2 below shows the antimicrobial activity of the methanol extract of cauliflower (part of the cauliflower except seeds).

Antimicrobial activity Number of bacteria (CFU / ml) Antibacterial activity (growth inhibition ring, mm) badge Shigella Flagmary - 3.78 X 10 ⁹ - SS Serratia Liquefax - 1.13 X 10 ¹⁰ - SS Salmonella Enteritidis + 9.62 X 10 ⁹ + S Salmonella Hedelberg ++ To X 10 ⁸ 14 SS Salmonella typhimurium ATCC14028 + 6.90 X 10 ⁹ + SS Salmonella typhimurium ATCC14023 - 4.90 X 10 ⁹ - SS Salmonella Zabiana - 1.25 X 10 ¹⁰ - SS E. coli O157: H7 + 6.35 X 10 ⁹ + MAC SCB4 - To X 10 ⁸ - PCA Takana - To X 10 ⁸ - PCA Bacillus - To X 10 ⁸ - PCA Enterobacter Kalkai - 1.89 X 10 ¹⁰ - MAC Staphylococcus aureus ++ To X 10 ⁶ 14 PCA Vibrio parahemoliticus ++ To X 10 ⁹ 16 TCBS

(-: No antimicrobial activity, +: <12 mm, ++:> 12 mm,

SS: Salmonella & Sigela Baby, MAC: MacConkey Baby, PCA: Plate Count Baby, TCBS: Vibrio Selected Baby)

In Table 2, the methanol extract except for the seed showed proliferative activity against Staphylococcus aureus, Vibrio parahemoliticus, Salmonella Hedelberg, and E. coliO157, and a clear clear zone was observed.

1 is a growth inhibitory ring in order to measure the antimicrobial activity of the extract of the medicinal herb, a is a medicinal herb leaf and eggplant methanol extract (1), herb leaf and eggplant extract (2) in Staphylococcus aureus (2) ), And the growth inhibitory ring formation was measured by placing a disk containing each of the methanol seed extract (3) and the seed number extract (4). b is the result of the disk test for Salmonella Heideberg, c is the Vibrio parahemoliticus.

Staphylococcus strains were inhibited by methanol extract except for seed, and vinegar extract, except seed (FIG. Growth is inhibited (Fig. 1b), Vibrio bacteria can be confirmed that the growth is inhibited by all four extracts. That is, since the extract of the vinegar has different antibacterial activity according to the extracting site or the solvent of the vinegar, the extract having the strongest antimicrobial activity against each bacterium or the extract in the form of a mixture is used for sterilization for a wide range of strains. Can be used. In addition, the extract of the present herb of the present invention has been confirmed that there is an excellent antimicrobial effect on Salmonella, Staphylococcus aureus, enteritis vibrio, which accounts for more than 50% of the food poisoning causative bacteria, can be used for the prevention of food poisoning.                     

Example 3: Thermal Stability Test of Sancho Extract

Methanol leaves and eggplant methanol extract was used as an antimicrobial material, and strains were confirmed for thermal stability using Vibrio parahemoliticus.

0.1 mL (10 ⁸ CFU / mL) of Vibrio parahemoliticus culture was plated on an LB plate, dried with sterile 10 mm paper discs, and then dried for 20 minutes at 30, 60, 67 and 90 ° C. For comparison experiments, 70% ethanol with inhibitory activity to bacteria was dropped on 70 ul of paper discs, respectively, and then dried at 37 ° C. for 24 hours to measure growth inhibition.

FIG. 2 shows that anti-Vibrio activity was compared with other commercially available disinfectants when the methanol extract was heat-treated at 60 ° C. (a) or 90 ° C. (b), respectively, (1) is methanol, and (2) Is Kamika-SD, (3) is Clinchol and (4) is ethanol methanol extract except seeds. In Figure 2, the diameter of the growth inhibiting ring of Vibrio parahemoliticus was confirmed to be about 18 mm regardless of the treatment temperature in all the heat treatment, it can be seen that the vinegar extract of the present invention has a thermal stability.

Example 4 Comparison of Antimicrobial Activity of Japanese Herb

Vibrio parahemolyticus, the causative agent of enteritis vibrio, was tested for antimicrobial activity against existing alcohol- and chlorine-based disinfectants.

0.1 mL of Vibrio parahemoliticus (~ X 10 ⁹ CFU / ml) was inoculated in TCBS agar and dried, and the paper disc was placed on the medium, followed by 64 ppm of 70% ethanol, Camica-SD and Sancho. Methanol extracts of tree leaves and branches were dropped by 70 ul, respectively, and cultured at 37 ° C. for 16 hours to observe the antimicrobial activity. As shown in Table 3, the antimicrobial activity was found in the order of 70% ethanol> vinegar leaves and eggplant methanol extract> Kamika-SD.

Sancho Extract 70% ethanol Kamika-SD Clear zone (cm) 1.2 1.8 0

In addition, it was confirmed that the synergistic effect of the methanol extract of the vinegar and eggplant of the present invention when used with the existing alcohol-based disinfectant.

0.1 mL of Vibrio parahemoliticus of the same bacterial count was inoculated into LB agar, dried, and then absorbed by 100 uL of a 70% ethanol mixture containing methanol extract of 10%, 1% and 0.1% on sterile paper discs. . The solvent was completely evaporated and placed on a test plate medium, incubated at 37 ° C. for 24 hours, and growth inhibition activity was observed.

Growth inhibition activity results are shown in Figure 3, the size of the growth inhibition ring is shown in Table 4 below. Figure 3 confirms the growth inhibition of Vibrio bacteria by methanol extract mixture except 70% ethanol and seeds, (1) is a 70% ethanol mixture containing 0.1% methanol extract, (2) 70% ethanol mixture containing 10% of the vinegar methanol extract, (3) is a 70% ethanol mixture containing 1% of the vinegar methanol extract, and (4) 70% ethanol.                     

Diameter of Inhibitory Ring (mm) 70% ethanol 100 ul 14 10% Japanese Herb Extract ^* 100 ul 17.5 1% Japanese Herb Extract 100 ul 17.5 0.1% Japanese Herb Extract 100 ul 18 Japanese Herb Extract 10 uL 12 ^* : Acid extract 100 ul + 70% ethanol 900 ul

In Table 4 and Figure 3, 70% ethanol when mixed with the methanol extract of the vinegar except for the seed can be seen about 2 times the inhibition ring increase regardless of the mixing ratio of the methanol extract.

Example 5 Determination of Minimum Inhibitory Concentration of Japanese Herb Extract

Minimal inhibitory concentrations of the extracts were tested using the Turbidmetric Assay method (Davidson, P. M and Parish, ME (1989) Food technology , 43, p148-155).

To the LB medium (10 ¹² to 10 ¹³ CFU / mL) inoculated with 0.1 mL of Vibrio parahemoliticus culture solution, methanol extract was added at a concentration of 0%, 0.5%, 3% and 10%, respectively. While culturing for 33 hours at ℃ ℃ absorbance at a wavelength of 600 nm using a spectrometer (Shimazu UV-1601) for each time zone test group. At this time, the number of viable cells was determined by inoculating the diluent solution by step into the 3M Petri-film and LB agar for general bacteria using the sample for each test group.

The results are shown in FIGS. 4 and 5.

Fig. 4 shows the incubation of Vibrio bacteria (A) inoculated in LB medium and Vibrio bacteria (B) inoculated in LB medium supplemented with ethanol methanol extract except for 10% seed, respectively. 5 is a graph showing the growth inhibition effect of Vibrio bacteria by concentration of methanol extract of vinegar except seeds. 4 and 5 it can be seen that methanol extract except for the seed inhibits the growth of Vibrio bacteria.

In addition, the growth inhibition of Vibrio bacteria of methanol extract except for seed was compared with the conventional alcohol disinfectants. As an alcohol-based disinfectant, Cincol commercially available from Jinro Co., Ltd. was used, and was carried out in the same manner as the above experiment. The results are shown in FIG.

FIG. 6 shows the growth of Vibrio bacteria in LB medium containing 10% of each of methanol extract, clean call and seeds of methanol extract / clean call (10% pepper, 90% clean call composition) except seeds. It is a graph observed. In Fig. 6, the methanol extract treatment group except the seed showed the best vibrio growth inhibitory activity, and the clean call treatment group and the acid / clean call treatment group also inhibited the growth of Vibrio, but the activity except the seed It was found to be weaker than the wood methanol extract alone treatment group.

Example 6: Preparation of ethanol extracts from Japanese vinegar and validation

Using ethanol as the extraction solvent was carried out in the same manner as in Example 1 to prepare a ethanol extract of Japanese vinegar. The vinegar ethanol extract is extracted from the part excluding the seed of the vinegar.                     

The antibacterial activity of Japanese ethanol extract was 70% ethanol, grapefruit extract (Killol-plus), alkaline ionized water, CG-100 (GCL), guard M-53 (Lotte), clean call (Jinro), Haikyu and Duo CleanX2. It was compared with (Duo clean).

0.1 mL of Vibrio parahemoliticus (~ X 10 ⁹ CFU / mL) was inoculated in LB agar, dried, and the paper discs were placed on the medium, and 50 ul of each of the ethanol extracts of ethanol and other disinfectants were dropped, and 37 ° C. Incubation at 16 hours was observed for inhibition. As a result, the antimicrobial activity was found in the order of vinegar ethanol extract ≒ 70% ethanol> grapefruit extract> Haikyu (Table 5).

Disinfectant Name Application concentration Vibrio parahemoliticus Clear zone (mm) Remarks Duo clean X 2 (50%) ~ X 10 ⁷ CFU / mL 0 Ztech Biomedical 70% Ethanol (Propanol-A) 95% 0 Japanese ethanol extract 10% 16 Grapefruit extract Stock solution 13 F Bank Alkaline ionized water Stock solution 0 Shin Gimpo Nonghyup CG 100 Stock solution 0 GCL corporation GuardM-53 Stock solution 11 Lotte Clean Call Stock solution 0 Course High-Q Stock solution 11

In other words, the ethanol extract of vinegar extracted with ethanol capable of food and beverage as a solvent has a significant antimicrobial activity as compared to the existing disinfectant, it can be confirmed that the utility as a food.

In addition, the minimum growth inhibition concentration of Vibrio bacteria was measured. According to the number of Vibrio parahemoliticus bacteria, 10%, 6%, 5%, 3%, and 1% ethanol was incubated with ethanol extract, and the absorbance was measured at 600 nm. As a result, Japanese pepper tree ethanol extract of 10%, 6%, 5% was inhibited up to ~ X 10 ⁴ to ~ X 10 parahaemolyticus growth of ⁷ CFU / ml, respectively, using the Japanese pepper tree ethanol extract in 1% and 3% When inhibiting the growth of Vibrio bacteria up to 10 ⁵ CFU / ml showed. Therefore, the addition content of the Japanese ethanol extract can be calculated as 5%.

As mentioned above, the vinegar extract of the present invention not only has a strong bactericidal effect against food poisoning bacteria but also is harmless to human body and can be widely used as a substitute for a conventional synthetic fungicide, and has excellent thermal stability and is easy for food processing. It can be applied to maintain the antimicrobial activity.

Claims (7)

Methanol methanol extract extracted with two or more portions selected from the group consisting of the bark of fruit, leaves, branches, roots, and stems of methanol; Contains ethanol aqueous solution as an active ingredient Salmonella Entebbe utility disk (Salmonella enteritidis), Salmonella hedel bug (Salmonella hedelberg), Salmonella typhimurium (Salmonella typhimurium) ATCC14028, and Vibrio para H. Molina T kusu (Vibrio parahaemolyticus) against one or more strains selected from the group consisting of antimicrobial Antimicrobial composition, characterized in that it has activity. delete delete The composition of claim 1, wherein the antimicrobial composition further comprises at least one antimicrobial substance selected from the group consisting of grapefruit extract, lysozyme, polylysine, protamine, cone albumin, avidin, and organic acids. delete Food comprising the antimicrobial composition according to claim 1 or 4 in 0.5 to 20% by weight. Food additives containing 0.5 to 20% by weight of the antimicrobial composition according to claim 1 or 4.

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