KR102228770B1 - Anti-microbial composition comprising rosemary extract and ascorbic acid as active ingredient - Google Patents

Abstract

The present invention relates to an antibacterial composition containing rosemary extract and ascorbic acid as an active ingredient, specifically rosemary extract; And at least one compound selected from ascorbic acid, calcium ascorbate, sodium ascorbate, and ascorbyl palmitate; an antibacterial composition containing as an active ingredient, and rosemary extract; And at least one compound selected from ascorbic acid, calcium ascorbate, sodium ascorbate, and ascorbyl palmitate; blended with an antibacterial object selected from foods, cosmetics, skin external preparations, pharmaceuticals and quasi-drugs to prevent microbial contamination of the antibacterial object Or how to sterilize it.
The antimicrobial composition of the present invention is made of a material that has been ingested for a long time by humans and has proven its safety, and can be safely used in products that are directly applied to the human body such as food, pharmaceuticals, cosmetics, tableware, cooking equipment, medical equipment, and beauty equipment. , By increasing the antibacterial activity of rosemary extract, it is economical to reduce the amount of rosemary extract used. In addition, according to the method of the present invention, it is possible to prevent and sterilize microbial contamination, which is economical and safe for food, pharmaceuticals, and cosmetics.

Description

Anti-microbial composition comprising rosemary extract and ascorbic acid as active ingredient}

The present invention relates to an antibacterial composition containing rosemary extract and ascorbic acid as an active ingredient, specifically rosemary extract; And at least one compound selected from ascorbic acid, calcium ascorbate, sodium ascorbate, and ascorbyl palmitate; an antibacterial composition containing as an active ingredient, and rosemary extract; And at least one compound selected from ascorbic acid, calcium ascorbate, sodium ascorbate, and ascorbyl palmitate; blended with an antibacterial object selected from foods, cosmetics, skin external preparations, pharmaceuticals and quasi-drugs to prevent microbial contamination of the antibacterial object Or how to sterilize it.

Food poisoning-related diseases and accidents are on the rise as the consumption of HMR products such as eating out and lunch boxes is increasing due to the diversification of dietary patterns due to the improvement of the income level according to the economic growth and the increase of the number of single households. In Korea, there have been accidents such as hamburger disease that caused hemolytic uremic syndrome, vibrio sepsis due to consumption of seafood, group food poisoning due to school and workplace meals, and such accidents occur every year. Due to the continuous increase in temperature due to the recent heatwave, interest in public hygiene and safety is increasing, and in particular, food poisoning problems for food and food products have emerged.

As food is changing in a variety of ways, it was necessary to extend the shelf life continuously so that it is safe and safe and does not affect the nutrition and taste of food, as well as problems of spoilage and contamination by various food-derived microorganisms such as bacteria, yeast and mold. Therefore, in the food industry, various antibacterial agents and preservatives are used to promote food preservation. Representatively, several chemically synthesized antibacterial agents such as sodium benzoate and potassium sorbate are used. However, these chemically synthesized antimicrobial agents are limited in the field and amount of use. In addition, as consumption of health-friendly and eco-friendly foods increases after the well-being craze, the need for new antimicrobial agents for natural foods is increasing due to a decrease in consumption due to a sense of resistance to chemically synthesized antimicrobial agents. In order to develop effective antimicrobial agents, it is necessary to understand and study various natural products at home and abroad. In particular, it is necessary to develop a natural antimicrobial agent that has a wide range of antibacterial and antimicrobial spectrum, an increase in antibacterial activity, and stability of formulation.

On the other hand, rosemary is an evergreen perennial plant belonging to the Lamiaceae family (Labiatae), and is widely used not only for edible but also for medicinal use, beauty, and fragrance. Rosemary extract contains many active substances such as rosemary acid, rosmanol, carnosic acid, and rosmarinic acid, and is known to have antibacterial activity against Gram-positive bacteria. , Anti-inflammatory, diuretic prevention, anti-aging, whitening, etc. It is widely used in the fields of medicine and cosmetics. In the current food industry, rosemary extract has been used to delay the oxidation process of fatty foods, improve the stability of foods, and increase the processing and storage time of foods by using strong antioxidant properties. However, the cost of such rosemary extract is very high, so if it is contained in large amounts in foods, there is a problem that the cost of the product increases.

In order to solve the above problems, the present inventor attempted to develop an antibacterial agent that can be safely used in food, while solving the problem of increasing product cost as a result by increasing the antibacterial activity of rosemary extract and reducing its amount.

Korean Patent Registration No. 10-0319252

FEMS Microbiology Letters. 224:247-253. Journal of Food Protection, 72(8): 1744-1752, 2009. Molecules. 17: 3989-4006.

Therefore, the main object of the present invention is to provide an antibacterial agent that can be safely used in foods, while solving the problem of increasing product cost as a result by increasing the antibacterial activity of rosemary extract and reducing its amount.

Another object of the present invention is to provide a method for preventing microbial contamination of foods, cosmetics, medicines, etc. safely at a lower cost by using rosemary extract.

According to one aspect of the present invention, the present invention is rosemary extract; And at least one compound selected from ascorbic acid, calcium ascorbate, sodium ascorbate, and ascorbyl palmitate; it provides an antibacterial composition containing as an active ingredient.

In the antimicrobial composition of the present invention, rosemary extract and ascorbic acid are contained in a weight ratio of 6.25:2,500 to 6.25:9,000, and the rosemary extract is contained in a concentration of 6.25 µg/ml or more, and the ascorbic acid is at a concentration of 2,500 µg/ml or more It contains as, and is preferably used for antibacterial use against Gram-positive bacteria.

In the antimicrobial composition of the present invention, rosemary extract and calcium ascorbate are contained in a weight ratio of 25: 3,200 to 25: 9,000, and the rosemary extract is contained in a concentration of 25 µg/ml or more, and the calcium ascorbate is 3,200 µg/ml It is contained in the above concentration, and is preferably used for antibacterial use against Gram-positive bacteria.

In the antimicrobial composition of the present invention, rosemary extract and sodium ascorbate are contained in a weight ratio of 12.5: 3,200 to 12.5: 9,000, and the rosemary extract is contained in a concentration of 12.5 µg/ml or more, and the sodium ascorbate is 3,200 µg/ml It is contained in the above concentration, and is preferably used for antibacterial use against Gram-positive bacteria.

In the antimicrobial composition of the present invention, rosemary extract and ascorbyl palmitate are contained in a weight ratio of 25: 3,200 to 25: 9,000, and the rosemary extract is contained in a concentration of 25 µg/ml or more, and the ascorbyl palmitate is 3,200 µg It is contained in a concentration of /ml or more, and is preferably used for antibacterial use against Gram-positive bacteria.

In the antimicrobial composition of the present invention, rosemary extract and ascorbic acid are contained in a weight ratio of 87.5:5,000 to 87.5:9,000, and the rosemary extract is contained in a concentration of 87.5 µg/ml or more, and the ascorbic acid is in a concentration of 5,000 µg/ml or more. It is contained as, and is preferably used for antibacterial use against Gram-negative bacteria.

According to another aspect of the present invention, the present invention is rosemary extract; And at least one compound selected from ascorbic acid, calcium ascorbate, sodium ascorbate, and ascorbyl palmitate; blended with an antibacterial object selected from foods, cosmetics, skin external preparations, pharmaceuticals and quasi-drugs to prevent microbial contamination of the antibacterial object Or provide a way to sterilize.

In the method of the present invention, it is preferable to prevent contamination or sterilize Gram-positive bacteria by mixing rosemary extract at a concentration of 6.25 to 40 µg/ml and ascorbic acid at a concentration of 2,500 to 9,000 µg/ml in the antibacterial object. .

In the method of the present invention, it is preferable to prevent contamination or sterilize gram-positive bacteria by mixing rosemary extract at a concentration of 25 to 40 µg/ml and calcium ascorbate at a concentration of 3,200 to 9,000 µg/ml in the antibacterial object. Do.

In the method of the present invention, it is preferable to prevent contamination or sterilize Gram-positive bacteria by mixing rosemary extract at a concentration of 12.5 to 40 µg/ml and sodium ascorbate at a concentration of 3,200 to 9,000 µg/ml in the antibacterial object. Do.

In the method of the present invention, it is possible to prevent or sterilize contamination of Gram-positive bacteria by mixing rosemary extract at a concentration of 25 to 40 µg/ml and ascorbyl palmitate at a concentration of 3,200 to 9,000 µg/ml in the antibacterial object. desirable.

In the method of the present invention, it is preferable to prevent contamination or sterilize Gram-negative bacteria by mixing rosemary extract at a concentration of 87.5 to 200 µg/ml and ascorbyl palmitate at a concentration of 5,000 to 9,000 µg/ml in the antibacterial object. desirable.

The antimicrobial composition of the present invention is made of a material that has been ingested for a long time by humans and has proven its safety, and can be safely used in products that are directly applied to the human body such as food, pharmaceuticals, cosmetics, tableware, cooking equipment, medical equipment, and beauty equipment. , It is economical because the amount of rosemary extract can be reduced by increasing the antibacterial activity of rosemary extract. In addition, according to the method of the present invention, economical and safe microbial contamination and sterilization of foods, pharmaceuticals, cosmetics, and the like can be prevented and sterilized.

The antimicrobial composition of the present invention contains, as an active ingredient, at least one compound selected from ascorbic acid, calcium ascorbate, sodium ascorbate, and ascorbyl palmitate together with rosemary extract, at a lower concentration than when using rosemary extract alone. It is characterized by showing excellent antibacterial and bactericidal effects.

The rosemary extract of the present invention can be obtained by extracting rosemary (scientific name: Rosmarinus officinalis L) as a raw material for extracting a plant that is commonly used. At this time, as the extraction solvent, an extraction solvent commonly used for plant extraction, such as distilled water, C1~C4 alcohol, acetone, ethyl acetate, chloroform, methylene chloride, or a mixed solvent thereof, can be used, preferably distilled water, ethanol , It is better to use alcohol or a mixed solution thereof. Preferably, it is good to use the leaves of rosemary as an extraction material. More preferably, after pulverizing rosemary leaves, it is recommended to use the one obtained by adding a 50 to 95% (v/v) ethanol solution of 5 to 10 times the weight of the pulverized product and extracting at 0 to 50°C for 5 hours or longer. It is good, and more preferably 7 to 9 times the weight of 70 to 85% (v/v) ethanol solution is added, and it is good to use the one obtained by shaking extraction at 30 to 40° C. for 10 hours or more. In addition, it is preferable to use after extraction by the above method, followed by filtration and concentration.

Ascorbic acid is known as vitamin C and is one of the important antioxidants. When ascorbic acid is used alone, it hardly exhibits antibacterial activity against microorganisms that are harmful to food, and even if it does, it exhibits only a weak effect at a high concentration of 10% or more. According to the present invention, it is possible to increase the antibacterial activity of rosemary extract by using together such ascorbic acid and a salt or analog thereof. Ascorbic acid has been developed and used in various salt forms, and the Ministry of Food and Drug Safety has six types such as sodium L-ascorbate, L-calcium ascorbate, L-ascorbyl stearate, and L-ascorbyl palmitate in a form that can be used as a food additive. It is registered.

According to the present invention, when rosemary extract and ascorbic acid are used in combination (combination 1), from a concentration of 6.25 µg/ml of rosemary extract and a concentration of 2,500 µg/ml of ascorbic acid, when a combination of rosemary extract and calcium ascorbate is used (combination 2 ) From the concentration of 25 μg/ml of rosemary extract and 3,200 μg/ml of calcium ascorbate, when using the combination of rosemary extract and sodium ascorbate (combination 3) concentration of 12.5 μg/ml of rosemary extract and 3,200 μg/ml of sodium ascorbate From the concentration of ml, when the rosemary extract and ascorbyl palmitate are used in combination (combination 4), excellent antibacterial activity against gram-positive bacteria can be exhibited from the concentration of 25 µg/ml of rosemary extract and 3,200 µg/ml of ascorbyl palmitate. . In addition, in the case of Combination 1, it can exhibit excellent antibacterial activity against Gram-negative bacteria from a concentration of 87.5 µg/ml of rosemary extract and 5,000 µg/ml of ascorbic acid. When the concentration of the rosemary extract or the concentration of the ascorbic acid compound is lowered at each concentration, the antimicrobial activity may be greatly reduced or may not be exerted, so the minimum concentration and the ratio of the two components can be said to be a very important factor in exerting the antibacterial activity.

Therefore, in the case of Combination 1 based on the above concentration, it is preferable to apply the concentrations of rosemary extract and ascorbic acid at a concentration of 6.25 to 40 µg/ml and 2,500 to 9,000 µg/ml, respectively, for antibacterial effect against Gram-positive bacteria. Preferably, it would be good to apply at a concentration of 6.25 to 20 µg/ml and 2,500 to 5,000 µg/ml, more preferably at a concentration of 6.25 to 10 µg/ml and 2,500 to 3,000 µg/ml, respectively. In addition, for the antimicrobial effect against Gram-negative bacteria in Combination 1, it is preferable to apply at a concentration of 87.5 to 200 µg/ml and 5,000 to 9,000 µg/ml, and more preferably 87.5 to 100 µg/ml and 5,000 to gram-negative bacteria, respectively. It would be good to apply at a concentration of 7,000 µg/ml, more preferably at a concentration of 87.5 to 90 µg/ml and 5,000 to 5,500 µg/ml, respectively. In the case of combination 2, it is preferable to apply the concentrations of rosemary extract and calcium ascorbate at a concentration of 25 to 40 µg/ml and 3,200 to 9,000 µg/ml, respectively, for antibacterial effect against Gram-positive bacteria, and more preferably 25 It would be good to apply at a concentration of ~ 35µg/ml, 3,200 ~ 6,000µg/ml, more preferably at a concentration of 25 ~ 30µg/ml and 3,200 ~ 4,000µg/ml, respectively. In the case of combination 3, it is preferable to apply the concentration of rosemary extract and sodium ascorbate at a concentration of 12.5 to 40 µg/ml and 3,200 to 9,000 µg/ml, respectively, for antibacterial effect against Gram-positive bacteria, and more preferably 12.5 each. It would be good to apply at a concentration of ~ 30µg/ml, 3,200 ~ 6,000µg/ml, more preferably at a concentration of 12.5 ~ 20µg/ml and 3,200 ~ 4,000µg/ml, respectively. In the case of Combination 4, it is preferable to apply the concentrations of rosemary extract and ascorbyl palmitate at a concentration of 25 to 40 µg/ml and 3,200 to 9,000 µg/ml, respectively, for antibacterial effect against Gram-positive bacteria, and more preferably each It would be good to apply at a concentration of 25 to 35 µg/ml and 3,200 to 6,000 µg/ml, more preferably at a concentration of 25 to 30 µg/ml and 3,200 to 4,000 µg/ml, respectively.

In addition, in the case of Combination 1 based on the above ratio, it is preferable to apply the ratio of rosemary extract and ascorbic acid in a weight ratio of 6.25:2,500 to 6.25:9,000 for antibacterial effect against Gram-positive bacteria, and more preferably 6.25:2,500 It would be good to apply the weight ratio of ~ 6.25: 5,000, more preferably 6.25: 2,500 ~ 6.25: 3,000 weight ratio. In addition, for the antibacterial effect against Gram-negative bacteria in Combination 1, it is preferable to apply it in a weight ratio of 87.5:5,000 to 87.5:9,000, more preferably in a weight ratio of 87.5:5,000 to 87.5:7,000, and more preferably 87.5: It would be better to apply in a weight ratio of 5,000 ~ 87.5: 5,500. In the case of combination 2, it is preferable to apply the ratio of rosemary extract and calcium ascorbate in a weight ratio of 25: 3,200 to 25: 9,000, and more preferably 25: 3,200 to 25: 6,000 for antibacterial effect against Gram-positive bacteria. As such, it would be better to apply it in a weight ratio of 25: 3,200 to 25: 4,000 more preferably. In the case of combination 3, it is preferable to apply the ratio of rosemary extract and sodium ascorbate in a weight ratio of 12.5: 3,200 to 12.5: 9,000, more preferably 12.5: 3,200 to 12.5: 6,000 for antibacterial effect against Gram-positive bacteria. As, more preferably, it would be good to apply in a weight ratio of 12.5: 3,200 to 12.5: 4,000. In the case of combination 4, it is preferable to apply the ratio of rosemary extract and ascorbyl palmitate in a weight ratio of 25: 3,200 to 25: 9,000 for antibacterial effect against Gram-positive bacteria, and more preferably 25: 3,200 to 25: 6,000. It would be good to apply in a weight ratio, more preferably in a weight ratio of 25: 3,200 to 25: 4,000.

The composition of the present invention can be applied by adding it to antibacterial objects such as foods, cosmetics, external preparations for skin, pharmaceuticals and quasi-drugs, and also machines or apparatuses such as tableware, kitchen equipment, beauty equipment, and medical equipment of the present invention. It can be applied by immersing in the composition or its diluent, or by spraying the composition of the present invention or its diluent on the surface of the device by these machines.

In addition, since the composition of the present invention can exhibit sterilizing power at the above concentration, it can be used as a sterilizing disinfectant. In particular, since all components are raw materials with low skin irritation, it can also be used as a sterilizing and disinfecting agent for skin of humans, companion animals and livestock.

In addition to each of the above components, the composition of the present invention may further contain other materials as long as it does not lower the antibacterial activity of the composition of the present invention. At this time, the material to be added is preferably a material that can be used in food or food additives.

The composition of the present invention may contain a solvent for dissolution or dispersion of each component. At this time, the solvent is preferably alcohol, water, or a mixture thereof. In addition, the composition of the present invention may be formulated in various forms acceptable pharmaceutically, food, or cosmetic.

The present invention also provides a method of preventing or sterilizing microbial contamination of the antimicrobial object by blending these two components into an antibacterial object based on the effect of the two components used as active ingredients of the composition of the present invention as described above. At this time, it is preferable to apply the blending concentration of the two components at the concentration suggested above.

In the method of the present invention, when the two components are mixed with the antibacterial object, it is preferable to use a mixer or a homogenizer to uniformly distribute the two components of the antibacterial object at the concentrations given above, and the antimicrobial object cannot be homogenized. In this case, it is preferable to ensure that the remaining homogenized portion is at the concentration given above.

Hereinafter, the present invention will be described in more detail through examples. Since these examples are for illustrative purposes only, the scope of the present invention is not to be construed as being limited by these examples.

[Example]

1. Manufacture of rosemary extract

After the dried rosemary leaves were pulverized with a blender, 70 to 85% (v/v) ethanol was added to 100 g of the pulverized product in an amount of 8 times, and extracted with shaking at 30 to 40° C. at 180 rpm for 12 hours.

The extract was filtered (Whatman No. 2 filter paper) to obtain an extract, and concentrated under reduced pressure to form powder.

2. Preparation of test solution

Ascorbic acid (S/N PHR1008), sodium ascorbate (S/N A7631), calcium ascorbate (S/N 1086356) and ascorbyl palmy Tate (Ascorbyl palmitate) (S/N 1043105) was purchased and used from Sigma.

Ascorbyl palmitate was dissolved in 100% ethanol, and ascorbic acid, sodium ascorbate, and calcium ascorbate were dissolved in purified water and used.

The rosemary extract of Example 1 was dissolved in 100% ethanol and used, and stored frozen at -20°C until the test.

The test solutions were used according to the set concentrations just before the minimum inhibitory concentration test and the fractional inhibitory concentration test.

3. Evaluation of antibacterial activity of rosemary extract and ascorbic acid compounds

As shown in Table 1 below, the antibacterial activity of the rosemary extract and four ascorbic acid compounds was evaluated. For the evaluation of antimicrobial activity, a method of measuring minimum inhibitory concentration (MIC), which is one of the methods for measuring antibacterial activity provided by CLSI (The Clinical & Laboratory Standards Institute), was used.

Test strains were E. coli ATCC 10536 and S. aureus ATCC 6538. First, the test strain was spread on TSA (Tryptic soy agar) medium and cultured at 36°C for 18 to 24 hours, and the cultured strain was inoculated in 10 ml of TSB (Triptic soy broth) medium and cultured at 36°C for 18 to 24 hours. After the culture was diluted with physiological saline, the number of viable cells ^{was adjusted to about 10 6} cfu/ml, which was then used as a test strain suspension.

The test solution prepared in Example 2 was added to the test strain suspension in which the number of viable cells was adjusted for each concentration, and the MIC of each material was measured.

Rosemary extract and ascorbyl palmitate dissolved in 100% ethanol were contained in less than 5% of the final volume for accurate results, so that antimicrobial activity by ethanol was not observed for each bacteria.

Based on the MIC results, in order to check the sterilization power, the solution of each MIC test group was inoculated on agar medium to perform a minimum bactericidal concentration (MBC) experiment, and the concentration of the material in the test group in which bacteria do not grow on the agar medium. MBC was determined by checking. The MBC results of each material are shown in Table 1.

Test substance MBC concentration (㎍ / ㎖) E. coli S. aureus Rosemary extract 350 50 Ascorbic acid 10,000 10,000 Calcium ascorbate >25,000 >25,000 Sodium ascorbate >25,000 >25,000 Ascorbyl Palmitate >10,000 >10,000

As a result of the antibacterial activity analysis, the MBC concentrations of rosemary extract and ascorbic acid for E. coli were 350 µg/ml and 10,000 µg/ml, respectively. For S. aureus , the MBC concentration of rosemary extract was 50 µg/ml, and the MBC concentration of ascorbic acid was 10,000 µg/ml, the same as that of E. coli. Rosemary extract showed stronger antibacterial activity against S. aureus. Except for ascorbic acid, the MBC concentration of the three other ascorbic acid compounds exceeded 10,000 ㎍/㎖ for both bacteria. In particular, calcium ascorbate and sodium ascorbate had a high concentration of 25,000 ㎍/㎖ for both bacteria. It appeared as MBC concentration.

Based on these results, a composite composition was prepared by combining each of four ascorbic acid compounds based on rosemary extract, which is a material with excellent antibacterial activity.

4. Evaluation of antibacterial activity of rosemary extract and ascorbic acid compound composite composition

A composite composition between the rosemary extract and the four ascorbic acid compounds was prepared. The composition of the prepared composite composition is shown in Table 2.

Composite composition extract Ascorbic acid Composition 1
Rosemary extract Ascorbic acid Composition 2 Ascorbyl Palmitate Composition 3 Calcium ascorbate Composition 4 Sodium ascorbate

The minimum sterilization concentration for the composite compositions 1 to 4 was evaluated. The test method used the method of experimenting by putting each antibacterial material in rows and columns using the form of a checkerboard. In the row direction, rosemary extract was diluted 2 times starting from the MBC concentration of 350 μg/ml, diluted 32 times, and applied at a concentration of 10.9 μg/ml, and in the column direction, one of the four ascorbic acid compounds. Was selected and diluted 2 times from the concentration of 10,000 µg/ml, which is the MBC concentration, and applied at a concentration of 312.5 µg/ml, which is the maximum 32-fold dilution. The concentration at the intersection point in each row and column was checked.

MBC was measured in the same manner as in Example 3. The results are shown in Table 3.

Experimental solution MBC concentration (㎍ / ㎖) E. coli S. aureus Rosemary extract Ascorbic acid Rosemary extract Ascorbic acid Composition 1 87.5 5,000 6.25 2,500 Composition 2 350 0 25 3,200 Composition 3 350 0 25 3,200 Composition 4 350 0 12.5 3,200

The antimicrobial activity of Compositions 1 to 4 was far superior to that of the rosemary extract and 4 types of ascorbic acid compounds.

In the case of E. coli , it showed excellent activity in Composition 1, which is a composite composition of rosemary extract and ascorbic acid, and the concentration of rosemary extract at this time was 1/4 times lower than that of a single treatment, and ascorbic acid was also 1/ It is twice the concentration.

In the case of S. aureus , all of the compositions 1 to 4 showed excellent activity at a much lower concentration than the concentration at the time of single treatment. In the case of Composition 1 with the best activity, rosemary extract showed activity at a concentration that is 1/8 times lower than that of a single treatment and ascorbic acid at a concentration that is 1/4 times lower. In the case of Compositions 2 and 3, activity was exhibited at 1/2 times lower concentration of rosemary extract, and in case of Composition 4 at 1/4 times lower concentration of rosemary extract. Except for ascorbic acid, the antimicrobial activity of the three ascorbic acid compounds was very low, with the MBC concentration exceeding 25,000µg/ml against the two bacteria. However, when treated with rosemary extract, the MBC concentration was 3,200µg/ml, showing a strong antibacterial activity. Was confirmed.

5. FBC evaluation of composite composition

Fractional bactericidal concentrations (Fractional Bactericidal Concentration, FBC) of the composite composition for the rosemary extract and four ascorbic acid compounds were evaluated. The fractional sterilization concentration value can be obtained by the method described in Equation 1 below.

[Equation 1]

In Equation 1, [A] is the concentration of compound A, MBC _A is the MBC value of compound A, and FBC _A is the MBC value for each test bacteria of compound A. Compound B is also applied in the same way as compound A. Index criteria were classified according to the criteria mentioned in Pei (2009)'s paper (see Table 4).

division shame Synergy <1 Partial synergy effect One Additive effect 1-2 Antagonism >2

The evaluation results of the FBC value are shown in Table 5.

Experimental strain Experimental solution FBC _Rosemary FBC _Ascorb FBC value division E. coli Composition 1 0.5 0.25 0.75 Synergy Composition 2 One 0 One Partial synergy effect Composition 3 One 0 One Partial synergy effect Composition 4 One 0 One Partial synergy effect S. aureus Composition 1 0.125 0.25 0.375 Synergy Composition 2 0.5 0 0.5 Synergy Composition 3 0.5 0 0.5 Synergy Composition 4 0.25 0 0.25 Synergy

Composition 1 showed a synergistic effect against E. coli. On the other hand, for S. aureus , the FBC values of all compositions were calculated to be 0.5 or less, and showed a high synergistic effect. Particularly, in the case of Composition 1, it was confirmed that the FBC value of the rosemary extract was 0.125, indicating that activity was exhibited at a concentration much lower than the MBC value when single treatment. It is confirmed that the value of FBC ascorb is 0 because the ascorbic acid compounds used in Compositions 2 to 4 did not show antibacterial and bactericidal activity against both bacteria, so the denominator value of Equation 1 is recognized as infinity and is expressed as 0. It was confirmed that rosemary extract and ascorbic acid compounds exhibited synergistic effects more than synergistic effects against E. coli and S. aureus , and particularly high synergistic effects against S. aureus, which are Gram-positive bacteria.

Claims (12)

Rosemary extract containing carnosic acid; And at least one ascorbic acid compound selected from ascorbic acid, calcium ascorbate, sodium ascorbate and ascorbyl palmitate; as an active ingredient,
An antibacterial composition for gram-positive bacteria, containing the rosemary extract at a concentration of 6.25 µg/ml or more and the ascorbic acid compound at a concentration of 2,500 µg/ml or more. The method of claim 1,
Containing the rosemary extract and the ascorbic acid in a weight ratio of 6.25: 2,500 to 6.25: 9,000,
Containing the rosemary extract at a concentration of 6.25 µg/ml or more and ascorbic acid at a concentration of 2,500 µg/ml or more,
Antibacterial composition against Gram-positive bacteria. The method of claim 1,
It contains the rosemary extract and the calcium ascorbate in a weight ratio of 25: 3,200 to 25: 9,000,
Containing the rosemary extract at a concentration of 25 μg/ml or more, and the calcium ascorbate at a concentration of 3,200 μg/ml or more,
Antibacterial composition against Gram-positive bacteria. The method of claim 1,
Containing the rosemary extract and the sodium ascorbate in a weight ratio of 12.5: 3,200 to 12.5: 9,000,
Containing the rosemary extract at a concentration of 12.5 μg/ml or more, and the sodium ascorbate at a concentration of 3,200 μg/ml or more,
Antibacterial composition against Gram-positive bacteria. The method of claim 1,
It contains the rosemary extract and the ascorbyl palmitate in a weight ratio of 25: 3,200 to 25: 9,000,
Containing the rosemary extract at a concentration of 25 ㎍ / ㎖ or more, the ascorbyl palmitate at a concentration of 3,200 ㎍ / ㎖ or more,
Antibacterial composition against Gram-positive bacteria. delete Rosemary extract containing carnosic acid at a concentration of 6.25 µg/ml or more; At least one ascorbic acid compound selected from ascorbic acid, calcium ascorbate, sodium ascorbate, and ascorbyl palmitate at a concentration of 2,500 µg/ml or more; A method for preventing or sterilizing contamination of Gram-positive bacteria of the antibacterial object by mixing it with an antibacterial object selected from food, cosmetics, skin external preparations, pharmaceuticals, and quasi-drugs. The method of claim 7,
The method characterized in that for preventing or sterilizing contamination of Gram-positive bacteria by mixing the rosemary extract at a concentration of 6.25 to 40 µg/ml and the ascorbic acid at a concentration of 2,500 to 9,000 µg/ml in the antibacterial object. The method of claim 7,
The method characterized in that for preventing or sterilizing contamination of Gram-positive bacteria by mixing the rosemary extract at a concentration of 25 to 40 µg/ml and the calcium ascorbate at a concentration of 3,200 to 9,000 µg/ml in the antibacterial object. The method of claim 7,
The method characterized in that for preventing or sterilizing contamination of Gram-positive bacteria by mixing the rosemary extract at a concentration of 12.5 to 40 μg/ml and the sodium ascorbate at a concentration of 3,200 to 9,000 μg/ml to the antibacterial object. The method of claim 7,
The method characterized in that for preventing or sterilizing contamination of Gram-positive bacteria by mixing the rosemary extract at a concentration of 25 to 40 µg/ml and ascorbyl palmitate at a concentration of 3,200 to 9,000 µg/ml in the antibacterial object. delete