KR20200015168A - Naturally occurring essential oil composition having antifungal activities against xerophilic fungi - Google Patents

Abstract

The present invention relates to an antifungal composition comprising natural essential oil. The composition according to the present invention can effectively inhibit the growth of xerotolerant fungi, and minimize the amount of essential oils used individually, thereby minimizing changes in sensory properties of foods when applied to the foods and reducing the cost by reducing the consumption of individual essential oil.

Description

Naturally occurring essential oil composition having antifungal activities against xerophilic fungi}

The present invention relates to a natural essential oil composition exhibiting antifungal activity against a dry fungus, a food packaging material comprising the same, a food packaging method, and a method for preventing or inhibiting growth of a dry fungus.

Penicillium penicillium Corruption in the 0.60 to 0.85 of the food): corylophilum) and Euro tium Les pence (a _w (water activity, Water Activity Eurotium naegeonseong fungi such as repens) is a relatively Microbiological Safety of Food is considered that the intermediate water (intermediate-moisture food) It is known to cause (Non Patent Literatures 1 to 3). Not only can P. corylophilum cause food rot, but research has also been reported to produce citrinin and fungal toxin as secondary products of metabolic processes (Non-Patent Document 4). In addition, penicillium and eurotium species are known to be common in the environment, so cross contamination with foods can easily occur.

Tolerant molds are generally resistant to a variety of external stresses, such as low pH, osmotic pressure, heat and high salt concentrations, as well as in dry environments, which can survive in foods and cause food poisoning in people who consume it. In order to control the growth of the dry-tolerant mold, a method of spraying ethanol or the like added to the food surface or adding a synthetic chemical preservative has been attempted, but it does not have a significant effect.

Recently, as health-oriented demands of consumers increase, there is a tendency to prefer natural antimicrobial agents such as essential oils rather than chemical synthetic substances as antimicrobial agents. Essential oils are highly volatile liquids extracted from plants and are known to have antimicrobial activity against various microorganisms. Essential oils having antimicrobial activity against these microorganisms can be used in the liquid or gaseous state. Essential oils can be used as an antimicrobial packaging material for foods, and have the advantage of high permeability to microorganisms contaminated in the gaps of food surfaces or food contact surfaces. This is increasing. However, the biggest limitation in applying natural essential oils to foods is that natural essential oils generally have a strong flavor and are likely to affect the sensory and physical properties of foods when used in foods. In addition, the use of a single type of essential oil alone has a disadvantage in that a large amount is required in order to exhibit an antimicrobial effect.

Accordingly, in order to treat natural essential oils in foods to control dry-tolerant molds, there is a need for a method capable of minimizing the amount of essential oils processed in foods or the amount of essential oils remaining in foods after processing.

Leistner, L., 1992. Food preservation by combined methods. Food Research International, 25 (2), 151-158. Suhr, K. I., & Nielsen, P. V., 2003. Antifungal activity of essential oils evaluated by two different application techniques against rye bread spoilage fungi. Journal of Applied Microbiology, 94 (4), 665-674. Pitt, J. I., & Hocking, A. D., 2009. The ecology of fungal food spoilage. In Fungi and food spoilage (pp. 3-9). Springer, Boston, MA. Dos Santos, C. M. C., Costa, G. L. D., & Figueroa-Villar, J. D., 2012. Identification of citrinin as the defence metabolite of Penicillium corylophilum stressed with the antagonist fungus Beauveria bassiana. Nat. Prod. Res. 26, 2316-2322.

SUMMARY OF THE INVENTION An object of the present invention is to provide an antifungal composition capable of minimizing the cost and change in organoleptic properties of foods by maintaining the antifungal effect of essential oils on dry tolerant fungi.

The present inventors found that the essential oil composition exhibiting antifungal activity against the dry fungus can be maximized antimicrobial synergy by searching for combinations and ratios of natural essential oils that cause antibacterial synergy during research on natural products. To complete.

The present invention prevents or inhibits the growth of dry fungus in food comprising the step of adding an antifungal composition against dry fungus comprising natural essential oil as an active ingredient, a food packaging material comprising the antifungal composition, and adding the antifungal composition to the food packaging material. To provide a food packaging method, the method for preventing or inhibiting the growth of dry-resistant mold in food comprising the step of treating the antifungal composition to food.

In the following examples, the combination of natural essential oils to confirm the antifungal activity against the dry fungus of the composition according to the ratio, it was confirmed that the amount of essential oils used separately from the antifungal synergy by the combination of natural essential oils. . Therefore, natural essential oils combined in certain proportions can be used as an active ingredient of the antifungal composition against dry fungus.

Hereinafter, the configuration of the present invention will be described in detail.

Cinnamon bark, citronella, may chang, gallic, thyme linalool, oregano, thyme thymol and spear It provides an antifungal composition for dry-tolerant fungus comprising two or more essential oils selected from the group consisting of mints as an active ingredient.

In the present invention, "essential oil" refers to an oily extract obtained by condensing a fragrance component from cinnamon shell, citronella, maychange, garlic, thyme linulol, oregano, thymemol or spearmint. In the preparation of the essential oil, steam distillation, solvent extraction using a volatile solvent (benzene, hexane, etc.), leaching (method of collecting the leaching sap by wounding the extraction target), compression method, supercritical extraction, etc. can be used. It is not limited thereto and may be prepared using a method known in the art. In addition, the essential oil may be a commercially available product. The essential oil can be applied as a liquid or a gas.

In one embodiment, the essential oil can be applied in gaseous state.

In the present invention, the antifungal essential oil composition exhibits antifungal activity against the dry fungus.

In the present invention, the dry-resistant mold means a mold that can grow even in relatively dry conditions, and refers to a mold that can be grown even in a water activity of 0.60-0.85.

In one embodiment, the dry resistant fungus may be Penicillium corylophilum or Eurotium repens .

The penicillium corypilum and eurotium lefen are fungi belonging to the green mold and the millet fungi, respectively, and may cause food decay in low or medium moisture foods. Penicillium corypilum can also produce fungal toxins that can cause food poisoning in people who consume it.

Therefore, the composition according to the present invention can prevent the degradation of food quality and food poisoning by inhibiting the growth of the fungi.

In the present invention, the antifungal composition may be one comprising a combination of two or more essential oils selected from the group consisting of cinnamon peel, citronella, maychange, garlic, thyme linalul, oregano, thymemol and spiramine.

In the present invention, the essential oil of the composition may include two essential oils are combined in a concentration ratio of 1: 0.1 to 1:32. For example, essential oil A and essential oil B have a concentration ratio of 1: 0.1 to 1:32, 1: 0.25 to 1:16, 1: 0.25 to 1: 8, 1: 0.5 to 1:16, 1: 0.5 to 1: 8, 1: 0.5 to 1: 4, 1: 1 to 1: 3, 1: 2 to 1: 4, 1: 2 to 1:16, 1: 4 to 1: 8 may be included in a combination ratio.

In one embodiment, the present invention is a cinnamon shell essential oil; And

It may be an antifungal composition comprising an essential oil selected from the group consisting of citronella, oregano, may chang, thyme linalul and garlic as an active ingredient.

In the above combination, cinnamon shell essential oil; And essential oils selected from the group consisting of citronella, oregano, may chang, thyme linalul and garlic. For example, the combination essential oil may be included in a concentration ratio of 1: 0.05 to 1:25, 1: 0.0625 to 1:20, and 1: 0.25 to 1:16.

In another embodiment, the present invention provides a citronella essential oil; And

It may be an antifungal composition comprising an essential oil selected from the group consisting of garlic, may chang, oregano, thymemol, spearmint and thyme linaule as an active ingredient.

In the combination, citronella essential oil; And essential oils selected from the group consisting of garlic, may Chang, oregano, thymemol, spearmint, and thyme linulul may be combined in a concentration ratio of 1: 0.01 to 1:32. For example, the combination essential oil may be included in a concentration ratio of 1: 0.05 to 1:16, 1: 0.1 to 1: 8, and 1: 0.125 to 1: 1.

In another embodiment, the present invention relates to a may Chang essential oil; And

It may be an antifungal composition comprising an essential oil selected from the group consisting of garlic, oregano and thyme linulol as an active ingredient.

In the combination, May Chang essential oil; And essential oils selected from the group consisting of garlic, oregano and thyme linulul may be combined at a concentration ratio of 1: 0.01 to 1:32. For example, the combination essential oil may be included in a concentration ratio of 1: 0.05 to 1:16, 1: 0.1 to 1: 8, and 1: 0.5 to 1: 1.

In another embodiment, the present invention is a garlic essential oil; And

It may be an antifungal composition comprising an essential oil selected from the group consisting of thyri linul, thymicol, oregano and spearamine as an active ingredient.

In the above combination, garlic essential oil; And essential oils selected from the group consisting of thyme linalul, thymicol, oregano, and spiramine may be combined at a concentration ratio of 1: 0.01 to 1:32. For example, the combination essential oil may be included in a concentration ratio of 1: 0.05 to 1:20, 1: 0.1 to 1:16, and 1: 2 to 1: 8.

In another embodiment, the present invention is a spearmint essential oil; And

It may be an antifungal composition comprising oregano or thyme linulol essential oil as an active ingredient.

In the above combination, spearmint essential oil; And oregano or thyme linulol essential oil may be combined in a concentration ratio of 1: 0.01 to 1:32. For example, the combination essential oil may be included in a concentration ratio of 1: 0.05 to 1:16, 1: 0.1 to 1: 8, and 1: 0.25 to 1: 2.

In another embodiment, the present invention may be an antifungal composition comprising thyrinal essential oil and thymicol essential oil as an active ingredient.

In the above combination, the time linarul essential oil and the time thymol essential oil are combined in a concentration ratio of 1: 0.01 to 1: 8, such as 1: 0.05 to 1: 4, 1: 0.1 to 1: 2, 1: 0.5 to 1: 1. It may be.

In the following examples, it was confirmed that the antifungal synergistic effect for the penicillium corypilum in the combination of cinnamon essential oil and citronella essential oil. Based on the above combination, cinnamon peel essential oil and citronella essential oil and May Chang essential oil, each of which showed a partial synergy effect, further confirmed the antifungal effect, and the combination of the three essential oils to penicillium corypilum It was confirmed that antifungal synergistic effects were observed.

Thus, the present invention may include an antifungal composition comprising cinnamon essential oil, citronella essential oil, and maychang essential oil as active ingredients.

In this combination, the cinnamon husk essential oil, citronella essential oil and maychang essential oil have a concentration ratio of 1: 0.5: 0.5 to 1: 4: 4, such as 1: 1: 1 to 1: 2: 2, 1: 2: 1 To 1: 2: 2.

In addition, according to the following examples, as a result of treating the food by combining two essential oils selected from the group consisting of cinnamon shell, citronella, garlic, may Chang, thyme mol, spearmint, thyme linulul and oregano, It was confirmed that there is an antifungal partial synergistic effect against Tium refense.

In the present invention, the content with respect to the total weight of the composition of the essential oil may vary depending on the concentration of the essential oil to be combined, the subject to which the composition of the present invention is applied.

On the other hand, the term 'comprising as an active ingredient' herein means containing an amount sufficient to achieve the antimicrobial efficacy or activity of the essential oil as described above. In the present invention, the essential oil may include 0.0001 μl / mL or more, such as 0.001 μl / mL or more, 0.004 μl / mL or more, but is not limited thereto.

In one embodiment of the invention, the essential oils combined in the composition of the present invention are, for example, when cinnamon peel essential oil and citronella essential oil are combined, the cinnamon peel essential oil is at least 0.0001 μl / mL, preferably 0.001 μl. / mL or more, more preferably 0.0015 μL / mL or more, and the Citronella essential oil contains 0.0001 μL / mL or more, preferably 0.001 μL / mL or more, and more preferably 0.0015 μL / mL or more.

In another embodiment of the invention, when May Chang essential oil is further combined with the combination, the May Chang essential oil is at least 0.0001 μL / mL, preferably at least 0.001 μL / mL, more preferably at least 0.0015 μL / mL. May be included.

Since the essential oil has no adverse effects on the human body even when excessively administered as a natural product, the upper limit of the amount of the essential oil included in the composition of the present invention can be carried out by those skilled in the art.

In addition, the present invention provides an antifungal food packaging material for inhibiting the growth of dry-resistant fungi comprising the antifungal composition. In one embodiment, the dry resistant fungus is Penicillium penicillium corylophilum ) or Eurotium repens .

According to the present invention, when the food packaging material is used, the amount of the essential oil is minimized than the amount of the essential oil used in the conventional food packaging material, thereby minimizing the organoleptic effect on the food and reducing the cost.

In addition, the present invention provides a food packaging method for inhibiting the growth of dry-resistant mold in food, comprising the step of applying or coating the antifungal composition on a food packaging material.

In the present invention, the step of applying or coating the antifungal composition on the food packaging material, Cinnamon bark (Cinnamon bark), Citronella (Citronella), May chang (Garlic), Garlic (Garlic), Thyme (Thyme) linalool), oregano, oregano, thyme thymol, and spearmint may further comprise combining the two or more essential oils selected from the group consisting of an antifungal composition.

Specific combinations of essential oils and antifungal compositions in the above method may equally apply or mutatis mutandis described above.

In addition, the present invention provides a method for preventing the growth of dry-tolerant mold in food, comprising the step of treating the antifungal composition to food.

In the present invention, the dry-resistant fungus may be Penicillium corylophilum or Eurotium repens .

In the present invention, the food may be a low water activity food or an intermediate-moisture food. The low moisture and medium moisture foods mean foods having a moisture content of about 1 to 20% by naturally occurring or by a drying or dehydration process. Food groups belonging to the low and medium moisture foods may be cereals (whole grains and whole grains), spices, spices, dry herbs, powdered foods, coffee, breads, confectionery, dried fruits, dried vegetables, seeds or beef jerky It is not limited.

In the present specification, the 'water activity' is the ratio (Ps / Po) of the water vapor pressure (P) of pure water to the water vapor pressure (Ps) indicated by the water in the food at any temperature. It is associated with chemical, biological and physical reactions in foods that change color, odor and flavor.

The low and medium moisture foods generally have a water activity of 0.60 to 0.85.

In the present invention, the step of treating the antifungal composition to food may be a step of spraying the composition on food.

The antifungal composition of the present invention not only effectively inhibits the growth of dry-tolerant mold, but also minimizes the amount of essential oils used individually due to antifungal synergies obtained by combining essential oils, thereby changing the sensory characteristics of foods when applied to foods. This can be minimized and the cost savings from reducing the use of individual refineries can also be achieved.

1 is a diagram illustrating a method for measuring the minimum inhibitory concentration (MIC) of essential oil gas for P. corylophilum .
Figure 2 is a schematic of the checkerboard assay method of cinnamon essential oil and citronella essential oil for P. corylophilum .
Figure 3 is a schematic diagram of the airtight container for confirming the antimicrobial activity of the combination of cinnamon peel essential oil, citronella essential oil and May Chang essential oil for P. corylophilum inoculated in beef jerky.

Advantages and features of the present invention and methods for achieving them will be apparent with reference to the embodiments described below in detail. However, the present invention is not limited to the embodiments disclosed below, but will be implemented in various forms, and only the embodiments are intended to complete the disclosure of the present invention, and the general knowledge in the technical field to which the present invention pertains. It is provided to fully convey the scope of the invention to those skilled in the art, and the present invention is defined only by the scope of the claims.

[ Example  One] Penicillium Corylophilum P. corylophilum )on  Determination of Minimum Inhibitory Concentration of Natural Refinery Gas

In order to determine the minimum inhibitory concentration (MIC) of natural essential oils against P. corylophilum , a gas antimicrobial activity measuring device and gas antimicrobial activity measurement method (Korean Patent No. 10-1406387) was used.

First, P. To prepare the extract of spore corylophilum, Dichloran 18% were incubated with P. corylophilum Glycerol agar (DG18) and the frequency divider 0.1% peptone water with 30% glycerol (0.1% PW30G) in 10 mL of the agar medium, The colonies on the culture medium containing the fungi were separated using a spreader, and then, a sterile gauze was used to filter the colony of P. corylophilum and the suspension of 0.1% PW30G. The spore count of the suspension was counted using a Neubauer counting chamber (hemocytometer) and a microscope. The spore suspension was diluted to a concentration of 10 ⁵ or 10 ⁶ spores / mL using 0.1% PW30G, and then mixed with 5 mL of the diluted spore suspension to prepare 10 mL of P. corylophilum spore extract for use in the experiment.

DG18 was prepared and autoclaved. Place 0.35 mL of the liquid medium in the upper container of the antimicrobial activity measuring device, and harden it. Dispense 10 μl of P. corylophilum spore extract (ca. 10 ⁶ spores / mL) to allow the microorganism to be adsorbed onto the medium. 30 minutes). Natural essential oils were binary diluted in stages using dimethyl sulfoxide (DMSO). After diluting the natural essential oil dilution by 10 μl into the lower container of the antimicrobial activity measuring device containing paper disc (Advantec Toyo Kaisha. Ltd., Tokyo, Japan), the upper container and the lower container were quickly combined and sealed. The sealed containers were incubated for 120 hours in a 25 ° C. incubator, and then visually observed colony formation on the medium to set the minimum concentration of essential oil gas without colony to minimum inhibition concentration (MIC). Figure 1 shows a method for measuring the MIC of the essential oil gas for P. corylophilum , the measurement results are shown in Table 1.

essential oil Minimal inhibitory concentration (µl / ml) Scientific name Common name Use area Allium sativum Garlic Root 0.0390 Cinnamomum zeylanicum Cinnamon bark Bark 0.1563 Litsea cubeba May chang Fruit 0.1563 Cymbopogon nardus Citronella Grass 0.3125 Thymus zygis  CT thymol Thyme thymol Leaf 0.3125 Origanum vulgare Oregano Leaf 0.6250 Mentha spicate Spearmint Flowering herb 0.6250 Thymus zygis  CT linalool Thyme linalool Flower 0.6250

As shown in Table 1, all of the garlic, cinnamon peel, maychange, citronella, thymemol, oregano, spearmint and thyme linul essential oils all exhibited low minimum inhibitory concentrations (MICs) of 0.6250 μl / mL or less . It was confirmed that the antifungal effect on corylophilum is excellent.

[ Example  2] Cinnamon  Peel essential oil gas Citronella  Refinery gas Combination  Antifungal Synergy Check

At present, since the experimental method for confirming the antibacterial synergy by combining natural essential oils is not standardized, a checkboard assay, which is a method of confirming the antimicrobial synergy by combining liquid antimicrobial substances, was used.

The antimicrobial activity measuring devices (vial type) are listed in 4 rows x 4 columns. Add 0.35 mL of DG18 agar (see Example 1) to the upper container of the antimicrobial activity measuring device and solidify for 30 minutes, and dispense 10 μl of P. corylophilum spore extract (10 ⁶ spores / mL) to adsorb the microorganisms onto the medium. It was maintained for 30 minutes in a biosafety hood (22 ± 2 ℃). An 8 mm diameter paper disc (Advantec Toyo Kaisha. Ltd., Tokyo, Japan) was placed in the lower container of the antimicrobial activity measuring device. Liquid refinery gases were sequentially diluted using DMSO until they reached 1 / 8th the concentration of the minimum inhibitory concentration (MIC) of the refinery gas. 2.5 μL of essential oil A, which was diluted from MIC concentration to 1/8 MIC concentration, was dispensed in the transverse direction (4 vials) of 16 vials, and binary diluted from MIC concentration to 1/8 MIC concentration in the longitudinal direction (4 vials). 2.5 μl of Essential Oil B was dispensed. Immediately after dispensing, the upper vessel and the lower vessel were quickly combined and sealed. The sealed containers were incubated for 120 hours in a 25 ° C. incubator, and then visually observed colony formation on the medium. In case colonies of P. corylophilum were not formed, fractional inhibitory concentration (FIC) and fractional inhibitory concentration index (FICI) were calculated to determine the antifungal synergy. At this time, if FICI is less than or equal to 0.5, synergistic effect, partial synergistic effect between 0.5 and 1, independent between 1 and 4, and antagonistic effect if 4 or more Judging there was. The said measuring method is shown in FIG. 2, and the measurement result is shown in Table 2. FIG.

[Equation]

Essential Oil A Refinery B Essential Oil Concentration (μl / mL) FIC FICI result Essential Oil A Refinery B Total refinery Essential Oil A Refinery B Cinnamon  Cinnamon bark Citronella (Citronella) 0.0390 0.0781 0.1171 0.2500 0.2500 0.5000 Synergy Garlic Citronella 0.0195 0.0390 0.0585 0.5000 0.1250 0.6250 Partial synergy Garlic Thyme linalool 0.0195 0.0781 0.0976 0.5000 0.1250 0.6250 Partial synergy May chang Cinnamon bark 0.0195 0.0781 0.0976 0.1250 0.5000 0.6250 Partial synergy May chang Citronella 0.0195 0.1563 0.1758 0.1250 0.5000 0.6250 Partial synergy Citronella Oregano 0.1563 0.0781 0.2344 0.5000 0.1250 0.6250 Partial synergy Cinnamon bark Oregano 0.0195 0.3125 0.3320 0.1250 0.5000 0.6250 Partial synergy Garlic Thyme thymol 0.0195 0.0390 0.0585 0.5000 0.2500 0.7500 Partial synergy Garlic May chang 0.0195 0.0390 0.0585 0.5000 0.2500 0.7500 Partial synergy Garlic Oregano 0.0195 0.1563 0.1758 0.5000 0.2500 0.7500 Partial synergy May chang Oregano 0.0781 0.1563 0.2344 0.5000 0.2500 0.7500 Partial synergy Garlic Spearmint 0.0195 0.1563 0.1758 0.5000 0.2500 0.7500 Partial synergy Citronella Thyme thymol 0.1563 0.0390 0.1953 0.5000 0.2500 0.7500 Partial synergy Citronella Spearmint 0.1563 0.1563 0.3125 0.5000 0.2500 0.7500 Partial synergy Citronella Thyme linalool 0.1563 0.1563 0.3125 0.5000 0.2500 0.7500 Partial synergy Thyme linalool Spearmint 0.3125 0.1563 0.4688 0.5000 0.2500 0.7500 Partial synergy

As shown in Table 2, synergistic or partial synergistic effects could be confirmed in various combination treatment groups of essential oil gases for P. corylophilum . In particular, the combination of cinnamon essential oil and citronella essential oil in a ratio of 1: 2 showed that the growth of P. corylophilum could be suppressed in a smaller amount than that of each essential oil alone. . In the combination treatment group, the FICI value was 0.5000, and it was confirmed that the combination of two essential oil gases had an antifungal synergistic effect against P. corylophilum .

[ Example  3] Cinnamon  Peel essential oil, Citronella  Refinery and may  Window of essential oil Combination  Confirm antimicrobial synergy

In Example 2, the antifungal synergistic effect of P. corylophilum was confirmed in the combination of cinnamon essential oil and citronella essential oil. Thus, the experiment was conducted by further combining the May Chang essential oil gas which showed a partial synergistic effect (FICI = 0.6250) in common when combined with cinnamon shell essential oil and citronella essential oil.

Antimicrobial activity measuring devices (64 vials) were listed in 4 rows x 4 columns x 4 sets. 0.35 mL of DG18 agar was added to the upper container of the antimicrobial activity measuring device and allowed to solidify for 30 minutes . Then , 10 μl of P. corylophilum spore extract (10 ⁶ spores / mL) was dispensed and the laminal flow biosafety hood (22 ± 2) was applied. 30 minutes). An 8 mm diameter paper disc (Advantec Toyo Kaisha, Ltd., Tokyo, Japan) was placed in the lower container of the antimicrobial activity measuring device. Liquid petroleum refinery gases were sequentially diluted from MIC of the refinery gas to 1/8 of the MIC using DMSO. 1.7 μl of essential oil A, which was diluted from MIC concentration to 1/8 MIC concentration, was dispensed in the transverse direction (4 vials) of 16 vials and binary diluted from MIC concentration to 1/8 MIC concentration in the longitudinal direction (4 vials). 1.7 μl of Essential Oil B was dispensed. Thus prepared four antibacterial activity measuring device set consisting of 16 vials. Thereafter, 1.7 μl of essential oil C diluted binary from MIC concentration to 1/8 MIC concentration was dispensed into each experimental instrument. Immediately after dispensing, the upper vessel and the lower vessel were quickly combined and sealed. The sealed containers were incubated in a 25 ° C. incubator for 120 hours and visually observed for colony formation on the medium. The antifungal synergistic effect of the essential oil gas combination was calculated by calculating FIC and FICI according to the same method as in Example 2. Confirmed. The results are shown in Table 3 below.

Essential Oil Concentration (μl / mL) FIC FICI result Cinnamon  skin Citronella may  window Total refinery Cinnamon  skin Citronella may  window 0.0195 0.0390 0.0195 0.0781 0.1250 0.1250 0.1250 0.3750 Synergy 0.0195 0.0390 0.0390 0.0975 0.1250 0.1250 0.2500 0.5000 Synergy 0.0195 0.0390 0.0781 0.1366 0.1250 0.1250 0.5000 0.7500 Partial synergy

In Example 2, the concentrations of cinnamon peel essential oil and citronella essential oil, which showed antifungal synergistic effects on P. corylophilum , were fixed, and the concentrations of the Maychang essential oil gas were different. When essential oils and May Chang essential oils were treated in combination in a ratio of 1: 2: 1 or 1: 2: 2, it was confirmed that the growth of P. corylophilum can be suppressed in a smaller amount than that of each essential oil. It was. FIC values of 0.3750 and 0.5000 in the combination treatment group, respectively, can be seen that there is an antifungal synergistic effect in the combination of three refinery gas.

[ Example  4] Confirmation of antifungal effect of essential oils in medium moisture foods

Cinnamon peel essential oil, citronella essential oil, maychang essential oil, and combinations that showed the lowest FICI in the previous example showed strong antibacterial activity against P. corylophilum (concentration ratio (volume ratio of 1: 2: 1)) The antimicrobial activity of was determined in P. corylophilum inoculated on beef jerky.

A sealed container for the experiment was prepared using a round polystyrene container. Figure 3 is a schematic diagram of a closed vessel manufactured to treat the refinery gas on the beef jerky surface. A small Petri dish lid was placed to place the jerky coupon inside the sealed container. The upper part of the refinery gases used to measure the MIC was inverted and placed next to the lid of a small Petri dish, and a sterile 8 mm paper disc was placed inside the upper part of the refinery frame. In order to maintain the internal relative humidity of the sealed container at 93%, 20 mL of saturated potassium nitrate was placed in the bottom of the sealed container at least 12 hours before the experiment, and then stored at 25 ° C.

After sterilizing the beef jerky coupon cut to 2 cm × 4 cm at room temperature (22 ± 2 ° C.), inoculate 10 points of 100 μl of P. corylophilum spore mixture (ca. 10 ⁵ spores / mL) on the jerky coupon. spot inoculation) and dried in a laminar flow biosafety hood (22 ± 2 ° C) for 1 hour. The dried jerky coupon was placed on the lid of a small Petri dish inside the sealed container, and the liquid refined gas was dispensed onto a paper disc. The amount of refined liquid oil dispensed was such that the final concentration of the refinery gas when it was all vaporized in a closed container was 0.0097, 0.0195, 0.0390 or 0.0781. For the essential oil gas combination treatment, the discs were dispensed onto the paper disc so that the concentrations of cinnamon shell essential oil, citronella essential oil, and maychang essential oil liquids were 1: 2: 1. After dispensing the liquid essential oil, the sealed container was sealed using parafilm and incubated at 25 ° C. for 120 hours.

Jerky coupons cultured with P. corylophilum were placed in a Whirl-pak bag containing 10 mL of 0.1% PW30G, and pummeled for 1 minute using a homogenizer. Decimal dilution of the jerky and 0.1% PW30G mixture using 0.1% PW30G was then plated onto DG18 agar medium. DG18 agar medium was incubated at 25 ° C. for 48 hours, and the colonies (dark green colonies) estimated to be P. corylophilum were counted and shown in Table 4 below. The theoretical limit of detection of plate smears was 0.1 log CFU / cm ² .

essential oil P. corylophilum  Number of objects (log CFU Of cm ² ) Initial value Essential Oil Concentration (μl / mL) 0 0.0097 0.0195 0.039 0.0781 Cinnamon  skin A2.9 ± 0.1c A5.2 ± 0.3a B4.4 ± 0.3b A4.5 ± 0.3b AB3.2 ± 0.4c B3.1 ± 0.3c Citronella A2.9 ± 0.1e A5.2 ± 0.3a A4.8 ± 0.2b B4.0 ± 0.3c A3.7 ± 0.5cd A3.6 ± 0.4d may  window A2.9 ± 0.1d A5.2 ± 0.3a A5.2 ± 0.3a B4.3 ± 0.2b AB3.5 ± 0.3c B2.9 ± 0.3d Cinnamon  skin, Citronella  Of May and May window A2.9 ± 0.1b A5.2 ± 0.3a A4.9 ± 0.1a C3.2 ± 0.0b B3.0 ± 0.4b B3.1 ± 0.3b

In the same column in Table 4, uppercase means that there is no significant difference ( P≤0.05 ), and lowercase means that there is no significant difference in the same row ( P≤0.05 ).

As shown in Table 4, the initial population of P. corylophilum inoculated on the jerky surface was ca. 2.9 log CFU / cm ² , the population was ca. Increased to 5.2 log CFU / cm ² . The minimum concentrations of each essential oil gas showed no significant difference from the initial population of P. corylophilum : cinnamon essential oil (0.0390 μl / mL), citronella essential oil (> 0.0781 μl / mL), and Maychang essential oil (0.0781 μl / mL) and three petroleum refinery gases (0.0195 μl / mL). At 0.0195 μl / mL, the population of P. corylophilum when the combination of cinnamon essential oil, citronella essential oil, and May Chang essential oil gas (concentration ratio (volume ratio of 1: 2: 1)) was treated, respectively. It was significantly lower ( P ≦ 0.05) compared to the population when treated with a single treatment. This result indicates that the combination of three refined gases can inhibit the growth of P. corylophilum in the jerky at lower concentrations than the single refined gas.

[ Example  5] Eurotium Lefence ( Eurotium repens )on  Determination of Minimum Inhibitory Concentration of Natural Refinery Gas

To determine the minimum inhibitory concentration (MIC) of the natural essential oil for E. repens, the experiment was performed in the same manner as in Example 1, except that instead of the E. repens P. corylophilum the first embodiment. The results are shown in Table 5 below.

essential oil Minimum inhibitory concentration ( MIC ) (Μl / mL) Scientific name Common name Use site Allium sativum Garlic Root 0.0390 Cinnamomum  zeylanicum Cinnamon bark Bark 0.1563 Cymbopogon nardus Citronella Grass 0.1563 Litsea cubeba May chang Fruit 0.1563 Origanum vulgare Oregano Leaf 0.1563 Mentha spicate Spearmint Flowering herb 0.3125 Thymus zygis  CT linalool Thyme linalool Flower 0.3125 Thymus zygis  CT thymol Thyme thymol Leaf 0.3125

As shown in Table 5, garlic, cinnamon husk, may chang, citronella, thyme thymol, oregano, spearmint and thyme linul essential oils all showed low MICs below 0.3125 μl / mL . It was confirmed that the antifungal effect on repens is excellent.

[ Example  6] Eurotium Lefence ( Eurotium repens )on  For refinery gas Combination  Identify antifungal synergies

In order to confirm the antifungal synergistic effect against Eurotium repens by combining natural essential oils, the experiment was carried out in the same manner as in Example 2. The results are shown in Table 6.

Essential Oil A Refinery B Essential Oil Concentration (μl / mL) FIC FICI result Essential Oil A Refinery B Total refinery Essential Oil A Refinery B Cinnamon bark May chang 0.0781 0.0195 0.0976 0.5000 0.1250 0.6250 part
Synergy Cinnamon bark Garlic 0.0781 0.0049 0.0830 0.5000 0.1250 0.6250 part
Synergy Cinnamon bark Citronella 0.0781 0.0195 0.0976 0.5000 0.1250 0.6250 Partial synergy Garlic Thyme thymol 0.0049 0.1563 0.1612 0.1250 0.5000 0.6250 Partial synergy Garlic May chang 0.0195 0.0195 0.0390 0.5000 0.1250 0.6250 Partial synergy Spearmint Thyme linalool 0.0390 0.1563 0.1953 0.1250 0.5000 0.6250 Partial synergy Cinnamon bark Thyme linalool 0.0781 0.0781 0.1563 0.5000 0.2500 0.7500 Partial synergy Garlic Spearmint 0.0195 0.0781 0.0976 0.5000 0.2500 0.7500 Partial synergy Garlic Thyme linalool 0.0097 0.1563 0.1660 0.2500 0.5000 0.7500 Partial synergy Garlic Oregano 0.0195 0.0390 0.0585 0.5000 0.2500 0.7500 Partial synergy May chang Thyme linalool 0.0781 0.0781 0.1563 0.5000 0.2500 0.7500 Partial synergy Oregano Spearmint 0.0390 0.1563 0.1953 0.2500 0.5000 0.7500 Partial synergy Thyme linalool Thyme thymol 0.1563 0.0781 0.2344 0.5000 0.2500 0.7500 Partial synergy

As shown in Table 6, the partial synergy effect was confirmed in the experimental group of various combinations of refinery gas for E. repens . Among them, the combination of cinnamon shells and maychang essential oils, the combination of cinnamon shells and garlic essential oils, the combination of cinnamon shells and citronella essential oils, the combination of garlic and thymol petroleum essential oils, the combination of garlic and maychang essential oils, and the spearmint and thyrina The combination of rule essential oils showed a FICI value of 0.6250, indicating a partial antifungal synergy.

Claims (15)

Cinnamon bark, Citronella, May chang, Garlic, Thyme linalool, Oregano, Thyme thymol and Spearmint Penicillium corypilum containing at least two essential oils selected from the group consisting of antifungal composition against corylophilum ) or Eurotium repens . The method of claim 1,
Cinnamon shell essential oil; And
Antifungal composition comprising an essential oil selected from the group consisting of citronella, oregano, may chang, thyme linarul and garlic. The method of claim 1,
Citronella essential oils; And
An antifungal composition comprising an essential oil selected from the group consisting of garlic, may chang, oregano, thymemol, spearmint and thyme linaule as an active ingredient. The method of claim 1,
Mei Chang essential oil; And
Antifungal composition comprising an essential oil selected from the group consisting of garlic, oregano and thyme linaule as an active ingredient. The method of claim 1,
Garlic essential oil; And
An antifungal composition comprising an essential oil selected from the group consisting of thyre linul, thymicol, oregano and spearamine as an active ingredient. The method of claim 1,
Spearmint essential oil; And
Antifungal composition comprising oregano or thyme linaul essential oil as an active ingredient. The method of claim 1,
An antifungal composition comprising thyme linul essential oil and thymicol essential oil as active ingredients. The method of claim 2,
The active ingredient is an antifungal composition comprising cinnamon essential oil and citronella essential oil. The method of claim 1,
The active ingredient is an antifungal composition comprising two essential oils in a concentration ratio of 1: 0.01 to 1:32. The method of claim 1,
An antifungal composition comprising cinnamon essential oil, citronella essential oil, and maychang essential oil as active ingredients. The method of claim 10,
Cinnamon peel essential oil, citronella essential oil and May Chang essential oil is an antifungal composition comprising a concentration ratio of 1: 0.5: 0.5 to 1: 4: 4. Penicillium corypilum containing the antifungal composition according to any one of claims 1 to 11. antifungal food packaging for corylophilum ) or Eurotium repens . Claims 1 to 11 of the penicillium corylophilum or Eurotium repens in food, comprising applying or coating the antifungal composition according to any one of claims 1 to 11 in a food packaging. Food packaging methods to prevent or inhibit growth. Penicillium corypilum in food, comprising the step of treating the antifungal composition according to any one of claims 1 to 11 to food. corylophilum ) or methods of preventing or inhibiting growth of Eurotium repens . The method of claim 14,
Wherein the food is a low-moisture food or a medium-moisture food in the method of preventing or inhibiting the growth of penicillium corypilum or eurotium lepens.

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