KR20110105580A - Antimicrobial and antifungal nelumbo nucifera gaertner and the manufacturing method thereof - Google Patents

Abstract

The present invention relates to an antibacterial and antifungal agent using the white lotus leaf extract and a method for producing the same. It relates to the antibacterial and antifungal agents used and the preparation method thereof.
The antibacterial and antifungal agent comprising the white lead leaf composite composition according to an aspect of the present invention is mixed with the white lead leaf composition and the plum extract in a ratio of 1: 4. In addition, the white lotus leaf composition is mixed with white lotus leaf extract, gallnut extract, small fennel extract in a ratio of 5: 2: 2.
Antibacterial and antifungal preparation method according to another aspect of the present invention is the extract of white lotus leaf extract, the extract of the galleng extract, the small fennel extract extraction step, the extract of plum extract, the production of white lead leaf composition, the production of white lead leaf composition Steps. The extract of white lead leaf extract extracts white lead leaf extract for 3 to 4 hours at 65 to 75 ° C. by adding ethanol of 65 to 75% to the white lead leaf. In the extracting step of the gall bladder extract 45-55% ethanol is added to the gall bladder to extract the gall bladder extract at 75 to 85 ° C. for 4 to 5 hours. The small fennel extract extraction step adds 45 to 55% of ethanol to the small fennel to extract the small fennel extract at 75 to 85 ℃ for 4 to 5 hours. The plum extract extraction step is to add water to the plum and pressurized for 2 to 3 hours to extract the plum extract. The white lead leaf composition manufacturing step is to produce a white lead leaf composition by mixing the white lead leaf extract, the gall bladder extract, and the small fennel extract in a ratio of 5: 2: 2. The white lead leaf composite composition manufacturing step comprises mixing the white lead leaf composition and the plum extract in a ratio of 1: 4 to create a white lead leaf composite composition.

Description

Antimicrobial and antifungal agent using white lotus leaf extract and its manufacturing method {Antimicrobial and antifungal nelumbo nucifera gaertner and the manufacturing method

The present invention relates to an antibacterial and antifungal agent using the white lotus leaf extract and a method for producing the same, and more particularly, the selected gallola extract, small fennel extract, and plum extract selected through the screening of white lotus leaf extract and antibacterial function extracted with ethanol as an extraction solvent. It relates to the antibacterial and antifungal agents used and the preparation method thereof.

Due to the development and industrialization of modern science and technology, simple processed foods are being mass produced and consumed, and the quality level is gradually increasing. In addition, frequent trade exchanges between countries due to globalization enable large-scale imports of cheap food ingredients from abroad, causing food deterioration and corruption during distribution and storage. This is mainly due to the action of microorganisms, and therefore health disorders caused by harmful microorganisms in food hygiene, that is, food poisoning, are increasing in importance as a large social problem.

Until now, it has been common to use bacteriostatic action and bactericidal action of harmful microorganisms that cause food poisoning, as well as disinfectants or preservatives of synthetic products. However, by denaturing and killing proteins of microorganisms causing decay by heat treatment such as boiling water and steam, it destroys toxins and parasites such as sporeworms and inactivates enzymes in foods to change foods over time. The sterilization method to prevent the food has improved storage, but has a disadvantage of freshness due to changes in the texture and taste. On the other hand, bactericidal action that changes the food itself into an environment where microorganisms are difficult to grow, improves its preservation or changes the preservation environment of food, which makes it difficult for microorganisms to proliferate. . In addition, excessive use of fungicides or preservatives of artificial synthetic products or a combination of various preservatives is known to cause side effects such as carcinogenesis and mutagenesis in addition to its intended function. Avoiding synthetic preservatives is emerging as a health-oriented consumer. Therefore, there is a growing demand from consumers for the development and use of natural preservatives that have no safety problems.

As a result, research on the antimicrobial activity of natural substances such as various edible plants and herbal medicines and researches to develop natural food preservatives are being actively conducted. The development and use of such natural antimicrobial materials is a good way to solve the negative side caused by the use of artificial synthetic preservatives, not to induce consumer avoidance, and to improve storage and safety.

Natural antimicrobial substances reported so far include animals, plants, proteins and enzymes, organic acids and bacteriocin, and various edible plants are known to contain considerable antimicrobial substances. Herbal medicines and edible plants, which have been used for a long time, are reported to have various physiological activities, and they can be added to foods without undergoing purification or pure separation process. There is an effect that can be obtained at the same time.

Nelumbo nucifera is a dicotyledon of the aquatic plants and is a dicotyledonous plant. It is native to southern Asia and North Australia, and is mainly grown in ponds and grown in paddy fields. Paleozoic plants widely distributed in tropical and temperate eastern Asia including Korea and China, including Korea and Japan. In Buddhism, sacred plants have been used as flowers, ornamentals and teas, and leaves and roots have been edible. The leaves in the herb are called lower lobe and are used for diarrhea, headache and dizziness, bleeding, postpartum blood treatment, enuresis, detoxification, etc. It contains dn-methylcoclaurine, liriodenine, tartaric acid, citric acid, malic acid, succinic acid and tannin.

Root and leaves of Nelumbo nucifera Gaertner have been used in folk medicine for metabolic diseases such as diabetes, hyperlipidemia and hypertension and are known to be effective for headache, bleeding and detoxification. In general, the roots and leaves of the lotus are used for medicinal and edible, especially lotus root has a lot of water-soluble fiber is known to be effective in reducing constipation and effective in lowering blood pressure. In addition, it has been found that there is a cholesterol lowering action because it contains the glycine sugar protein, and tannin contained in the lotus root has a strong astringent action and is known to be excellent in hemostatic effect and excellent in antioxidant activity.

An object of the present invention is to provide a natural antimicrobial material using white lead leaf that can solve the negative side caused by the use of artificial preservatives and not only to avoid the avoidance of consumers, but also to improve the shelf life and safety.

The antibacterial and antifungal agent comprising the white lead leaf composite composition according to an aspect of the present invention is mixed with the white lead leaf composition and the plum extract in a ratio of 1: 4. In addition, the white lotus leaf composition is mixed with white lotus leaf extract, gallnut extract, small fennel extract in a ratio of 5: 2: 2.

Antibacterial and antifungal preparation method according to another aspect of the present invention is the extract of white lotus leaf extract, the extract of the galleng extract, the small fennel extract extraction step, the extract of plum extract, the production of white lead leaf composition, the production of white lead leaf composition Steps. The extract of white lead leaf extract extracts white lead leaf extract for 3 to 4 hours at 65 to 75 ° C. by adding ethanol of 65 to 75% to the white lead leaf. In the extracting step of the gall bladder extract 45-55% ethanol is added to the gall bladder to extract the gall bladder extract at 75 to 85 ° C. for 4 to 5 hours. The small fennel extract extraction step adds 45 to 55% of ethanol to the small fennel to extract the small fennel extract at 75 to 85 ℃ for 4 to 5 hours. The plum extract extraction step is to add water to the plum and pressurized for 2 to 3 hours to extract the plum extract. The white lead leaf composition manufacturing step is to produce a white lead leaf composition by mixing the white lead leaf extract, the gall bladder extract, and the small fennel extract in a ratio of 5: 2: 2. The white lead leaf composite composition manufacturing step comprises mixing the white lead leaf composition and the plum extract in a ratio of 1: 4 to create a white lead leaf composite composition.

According to the present invention by providing an antibacterial and antifungal agent using the white lotus leaf extract, it is possible to solve the negative side caused by the use of artificial synthetic preservatives, it is possible to prevent the avoidance phenomenon from the consumer. In addition, it can solve the storage problems and safety problems of existing antibacterial and antifungal agents.

1 is a flow chart of an embodiment of a method for producing an antibacterial and antifungal agent comprising a white lead leaf composite composition according to the present invention.

One embodiment of the antibacterial and antifungal agent comprising the white lead leaf composite composition according to the present invention is mixed with the white lead leaf composition and plum extract in a ratio of 1: 4. The white lotus leaf composition, the white lotus leaf extract, the gall bladder extract, the small fennel extract is mixed in a ratio of 5: 2: 2.

 The antimicrobial and antifungal preparations include white lead leaf extract extraction step (S10), five-leaf extract extract step (S20), small fennel extract extraction step (S30), plum extract extraction step (S40), and white lead leaf composition manufacturing step. (S50), and white lead composite composition manufacturing step (S60).

White leaf extract extract step (S10) by adding about 70% ethanol 10 times to the white leaf and extract the white leaf extract for 4 hours at 70 ℃.

In the gall extract extract step (S20), 50% ethanol is added about 10 times to the gall extract to extract the gall extract for 5 hours at 80 ° C.

In the small fennel extract extraction step (S30), 50% of ethanol is added to the small fennel by about 10 times to extract the small fennel extract at 80 ° C. for 5 hours.

In the plum extract extraction step (S40) by adding about 5 times the water to the plum and pressurized for 3 hours to extract the plum extract.

White lead leaf composition manufacturing step (S50) is mixed with the white lead leaf extract, the gall bladder extract, and the small fennel extract in a ratio of 5: 2: 2 to make a white lead leaf composition.

 White lead leaf composite composition manufacturing step (S60) by mixing the white lead leaf composition and the plum extract in a ratio of 1: 4 to produce a white leaf composite composition of antibacterial and antifungal agents.

Hereinafter will be described an experiment to determine the antimicrobial activity of the white lotus leaf composite composition.

The white lotus leaf used in this study was harvested from July to September. It was supplied from Hamyang Sangrim Farming Cooperative in Hamyang-gun and used for frozen storage. The gallola, Sohhoe, and plum were directly purchased from cultivated farms or herbal medicines.

In this experiment, antibacterial and antifungal activity was measured by using the white lotus leaf extract and the white lotus leaf composite composition.

White lotus leaf extract extract

First, white lotus leaf extract was extracted. Water, 30% ethanol, 50% ethanol, 70% ethanol was added to 200 g of white lotus leaf, and extracted at 60 ° C. for 5 hours, filtered using filter paper (Whatman No. 2), followed by rotary evaporator (Eyela 2Cs-50, Rikakikai Co., Tokyo, Japan) was concentrated under reduced pressure to obtain a hot water extract and an ethanol extract, the extraction yield of each solvent was measured by 105 ℃ drying method.

After selecting a solvent with excellent extraction yield, extraction was performed at different extraction temperature and time under the same conditions of 200 g of white lead leaf and 2 L of solvent, filtered using filter paper (Whatman No. 2), and then rotary evaporator (Eyela 2Cs- 50, Rikakikai Co., Tokyo, Japan) was concentrated under reduced pressure at 50 ℃ or less to obtain an extract according to the conditions, each extraction yield was measured by 105 ℃ drying method.

In the optimum extraction conditions determined by the above process, the extraction yield during standing and stirring was compared and measured using a 105 ° C. drying method.

 Extraction Yield Results by Solvent Concentration

30, 50, 70% ethanol with different concentrations of water was added to 200 g of white lotus leaf and extracted at 60 ° C for 5 hours, and the extraction yields of hot water extract and ethanol extract by concentration were measured using 105 ° C drying. As shown in Fig. 1, the extraction yield of 70% ethanol as the solvent was the highest at 6.05%. Where% yield is solid in extract or fraction (g) / raw material (g) × 100.

density(%) Extraction yield (%) Water
30% ethanol
50% ethanol
70% ethanol 3.52
4.66
4.92
6.05

 Extraction yield according to extraction temperature and time

In the results of Table 1, 200 g of white lotus leaf was extracted by using 70% ethanol, which showed the highest extraction yield, as a solvent. As a result of measuring the extraction yield of each condition by 105 ℃ drying method, the extraction yield was the highest when extracted at 80 ℃ for 6 hours. However, the extraction yield was not high compared with the extraction for 4 hours at 70 ℃, and considering the energy consumed to extract, the best in terms of efficiency was 6.25% when extracted for 4 hours at 70 ℃.

Temperature Extraction yield (%) 2 hours 4 hours 6 hours 50 ℃
60 ℃
70 ℃
80 ℃ 3.79
6.02
6.03
6.02 3.80
6.0
6.25
6.27 3.84
6.05
6.27
6.28

Extraction yield with standing and stirring

When 70% ethanol, which is the optimum extraction condition identified in the results of Table 1 and Table 2, was extracted for 4 hours at 70 ° C with a solvent, the extraction yields during standing and stirring were compared. The extraction yield was more than 1.2 times better.

Extraction Type Extraction yield (%) Stationary
Shaking 5.20
6.25

Ginseng Extract, Small Fennel Extract, Plum Extract

Extracting quintet, small fennel and plums selected as antimicrobial material, added 10 times 50% ethanol to each quince, small fennel for 5 hours at 80 ℃ and filtered using filter paper (Whatman No.2). Next, the mixture was concentrated under reduced pressure with a rotary evaporator (Eyela 2Cs-50, Rikakikai Co., Tokyo, Japan), and the plums were extracted by pulverizing only the pulp part after washing and pressing and leaching for 3 hours by adding 5 times the water to the sample. The filtrate was filtered using filter paper (Whatman No. 2), concentrated under reduced pressure with a rotary evaporator (Eyela 2Cs-50, Rikakikai Co., Tokyo, Japan), and centrifuged to remove impurities.

Antibacterial and antifungal activity measurement

Use strain and medium

The strains used in the antibacterial and antifungal experiments are microorganisms involved in food decay, decay, and hygiene, as shown in Table 4. Gram-positive bacteria include Staphylococcus aureus ATCC 25923, which causes enterotoxin to cause food poisoning, Bacillus subtilis ATCC 6633, which is involved in the decay of protein foods, Bacillus cereus ATCC 9634, which is widely distributed in nature, causing food deterioration, Streptococcus mutans ATCC 25175, Gram-negative bacteria include Escherichia coli ATCC 23736, an indicator bacterium of food hygiene contamination, Salmonella typhimurium ATCC 14028, a pathogen of common infectious diseases, Pseudomonas aeruginosa ATCC 25923, a virulence bacterium that causes bacterial food poisoning, Vibrio parahaemolyticus ATCC As a fungus, Aspergillus oryzae ATCC 9362, Aspergillus niger ATCC 10254, Aspergillus flavus ATCC 10124, Trichophyton mentagrophytes KCTC 6316 were used. The strain was used by the Korea Institute of Bioscience and Biotechnology (Biological Resource Center) Gene Bank (Korean Collection for Type Cultures, KCTC), and was used in Staphylococcus aureus , Bacillus subtilis , Bacillus cereus , Escherichia coli , and Salmonella typhimurium. , Pseudomonas aeruginosa strains were inoculated with platinum and inoculated in nutrient broth (NB, Difco, USA) medium at 37 ° C, Vibrio parahaemolyticus was inoculated in marine broth (MB, Difco, USA) medium at 30 ° C, Streptococcus mutans was brain heart infusion After inoculation in (BHI, Difco, USA) medium, the cells were incubated for 24 hours in an incubator at 37 ° C. Aspergillus oryzae, Aspergillus niger, Aspergillus flavus, Trichophyton mentagrophytes were inoculated in potato dextrose agar (PDA, Difco, USA) medium. It was used by incubating for 48 to 60 hours in the incubator ℃.

Strains ATCC / KCTC NO. Gram positive bacteria Staphylococcus aureus
Bacillus subtilis
Bacillus cereus
Streptococcus mutans 25923/1621
6633/1021
9634/1012
25175/3065 Gram negative bacteria Escherichia coli
Salmonella typhimurium
Pseudomonas aeruginosa
Vibrio parahaemolyticus 23736/1041
14028/2515
27853/2004
17802/2729 Fungi Aspergillus oryzae
Aspergillus niger
Aspergillus flavus
Trichophyton mentagrophytes 9362/6377
10254/6906
10124/6143
-/ 6316

Determination of Antimicrobial and Antifungal Activity of Extracts from White Leaf and White Leaf

Paper disc agar diffusion was performed to determine the antimicrobial activity of white lotus leaf extract and white lotus leaf composite composition. Bacteria Staphylococcus aureus , Bacillus subtilis , Bacillus cereus , Escherichia coli , Salmonella typhimurium , Pseudomonas aeruginosa are nutrient broth (NB, Difco, USA) medium, Vibrio parahaemolyticus is marine broth (MB, Difco, USA) medium, Streptococcus mutans is brain (BHI, Difco, USA) Inoculated with 8 kinds of bacteria in the medium and incubated for 9 to 12 hours, the cells were harvested by centrifugation (7,000 rpm, 20 min) activated cells. The collected cells were diluted with sterile water and adjusted to an absorbance of 0.5 at 600 nm using a UV spectrophotometer (Genesys 10UV, Thermo., USA), and 0.1 ml of each suspension was added to nutrient agar medium ( Vibrio parahaemolyticus was used for marine agar medium, Streptococcus mutans were spread evenly over the brain heart infusion agar medium) and then absorbed the white lotus leaf extract and the white lotus leaf composite composition to a concentration of 0.5, 1, 2, 4 mg / disc on each plate medium (Φ 8 mm). , Advantec Co.) was incubated for 24 hours in a 37 ℃ incubator ( Vibrio parahaemolyticus 30 ℃), and the presence and size of the clear zone around the paper disc was measured.

In order to measure the antifungal activity of the white lotus leaf extract and the white lotus leaf composite composition, the mycelia of each fungus were sterilized with sterile gauze and sterilized so that the absorbance was 0.5 at 600 nm using a UV spectrophotometer (Genesys 10UV, Thermo., USA). After diluting with water, inoculating 0.1 ml each of potato dextrose agar (PDA, Difco, USA) medium, and coagulating by pouring PDA medium before solidification. A sterilized paper disc (Φ 8 mm, Advantec Co.), which absorbed white lead leaf extract and white lead leaf composition to concentrations of 0.5, 1, 2, and 4 mg / disc, was placed on a plate medium containing test bacteria, and 30 ° C. After incubation for 24 to 48 hours, the presence and size of the clear zone around the paper disc were measured.

The antibacterial and antifungal activity was measured by paper disc agar diffusion method using a sample of white lead leaf extract and white lead leaf composite composition after heat treatment at 100 ° C. for 30 minutes and at 121 ° C. for 15 minutes.

Two Gram-positive bacteria ( Bacillus cereus, Streptococcus mutans ), two Gram-negative bacteria ( Escherichia coli, Vibrio parahaemolyticus ), and two fungi ( Aspergillus oryzae, Trichophyton mentagrophytes ) were selected from the strains in Table 4. Centrifuged (7,000 rpm, 20 min), and the microbial suspension obtained by diluting the collected cells with sterile water and the spore suspension with fungal mycelium sterilized with sterile gauze were prepared using UV spectrophotometer (Genesys 10UV, Thermo., USA). Adjust the absorbance to 0.5 at nm. 0.1 ml of each suspension and spore suspension are evenly spread on a suitable plate medium, and a paper disc (Φ 8 mm, Advantec Co.) is placed on the surface of each plate medium, followed by 4 mg / Absorption at the disc concentration was carried out for 24 to 48 hours at the appropriate temperature (bacteria 37 ℃, Vibrio parahaemolyticus and fungi 30 ℃) and the presence and size of the clear zone around the paper disc was measured.

Growth inhibition activity of the white lotus leaf composite composition was measured by the liquid medium dilution method. Bacteria Staphylococcus aureus , Bacillus subtilis , Bacillus cereus , Escherichia coli , Salmonella typhimurium , Pseudomonas aeruginosa are nutrient broth (NB, Difco, USA) medium, Vibrio parahaemolyticus is marine broth (MB, Difco, USA) medium, Streptococcus mutans is brain (BHI, Difco, USA) Inoculated each bacterial solution in the medium and incubated for 9 to 12 hours at 37 ℃ ( Vibrio parahaemolyticus 30 ℃), and then centrifuged (7,000 rpm, 20 min) the activated bacteria solution and the collected cells Dilution with sterile water was used to adjust the absorbance to 0.5 at 600 nm using a UV spectrophotometer (Genesys 10UV, Thermo., USA). White lead leaf composite composition adjusted to 0, 25, 50, 100, 200, 400, 800, 1,600 ppm (0, 0.025, 0.05, 0.1, 0.2, 0.4, 0.8, 1.6 mg / ml) 50 μl of each bacterial suspension was inoculated into 950 μl of the appropriate liquid medium mixture and incubated at the appropriate temperature for 24 hours, and then absorbance was measured at 600 nm to calculate the growth inhibition effect.

The mycelia of fungi Aspergillus oryzae , Aspergillus niger , Aspergillus flavus and Trichophyton mentagrophytes were sterilized with sterile gauze and diluted with UV spectrophotometer (Genesys 10UV, Thermo., USA) to absorbance at 600 nm to 0.5 and then white lead leaf complex. White leaf composite composition and appropriate liquid medium whose composition is adjusted to 0, 25, 50, 100, 200, 400, 800, 1,600 ppm (0, 0.025, 0.05, 0.1, 0.2, 0.4, 0.8, 1.6 mg / ml) 50 μl of each spore suspension was inoculated into 950 μl of the mixed solution and incubated at 30 ° C. for 24 hours, and then visually determined minimal inhibition concentration (MIC) with significant inhibition.

Results and Discussion

Antibacterial and Antifungal Activity of Baekyeonyeon Extract

Smear the bacterial suspension and spore suspension of each activated test strain on a suitable plate medium, and load the paper disc which adsorbed 70% ethanol extract of white lotus leaf to 0.5, 1, 2, 4 mg / disc concentration. ℃, Vibrio parahaemolyticus and fungi 30 ℃) after incubation for 24 hours was measured the presence and size of the clear zone around the paper disc was as shown in Table 5. Bacillus cereus, Aspergillus oryzae, Escherichia coli strains showed clear zone at 1 mg / disc concentration, while Streptococcus mutans, Vibrio parahaemolyticus, Staphylococcus aureus, Trichophyton mentagrophytes strains showed clear zone at 2 mg / disc concentration. Activity was shown. Clear zones were observed at 4 mg / disc for Bacillus subtilis and Salmonella typhimurium strains, and at 4 mg / disc for Pseudomonas aeruginosa, Aspergillus niger and Aspergillus flavus .

Inhibition zone (mm) 0.5mg / disc 1mg / disc 2mg / disc 4mg / disc Staphylococcus aureus
Bacillus subtilis
Bacillus cereus
Streptococcus mutans
Escherichia coli
Salmonella typhimurium
Pseudomonas aeruginosa
Vibrio parahaemolyticus
Aspergillus oryzae
Aspergillus niger
Aspergillus flavus
Trichophyton mentagrophytes -
-
-
-
-
-
-
-
-
-
-
- -
-
++
-
+
-
-
-
++
-
-
- +
-
++
++
++
-
-
++
++
-
-
+ ++
++
+++
+++
+++
+
-
++
+++
-
-
++

Where + is less than 10 mm, ++ is 10 to 13 mm, and +++ is 14 to 17 mm.

The antibacterial and antifungal activity of six strains was measured by paper disc agar diffusion method using a sample heat-treated with 70% ethanol extract of white lotus leaf at 100 ° C for 30 minutes and 121 ° C for 15 minutes. Even after strong heat treatment, the antibacterial activity of 70% ethanol extract of white lotus leaf was almost unchanged.

Inhibition zone (mm) 100 ℃, 30min 121 ℃, 15min Escherichia coli
Bacillus cereus
Vibrio parahaemolyticus
Streptococcus mutans
Aspergillus oryzae
Trichophyton mentagrophytes +++
+++
++
+++
+++
++ +++
+++
++
+++
+++
++

Where + is less than 10 mm, ++ is 10 to 13 mm, and +++ is 14 to 17 mm.

 Antimicrobial and Antifungal Activity Results of Baekyeon Leaf Composite Compositions

Smear the bacterial suspension and spore suspension of each activated test strain on a suitable plate medium, and load the paper disc adsorbed so that the white lotus leaf composite composition is 0.5, 1, 2, 4 mg / disc concentration. Vibrio parahaemolyticus and fungi 30 ℃) after incubation for 24 hours, the presence and size of the clear zone generated around the paper disc was measured as shown in Table 7. Escherichia coli, Salmonella typhimurium strains showed clear zone at 0.5 mg / disc concentration, and Trichophyton mentagrophytes, Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, Vibrio parahaemolyticus, Aspergillus oryzae, Streptococcus mutans strains at 1 mg / disc concentration. It showed strong antibacterial and antifungal activity, and clear zone was appeared at 4 mg / disc concentration for Bacillus subtilis strain. Aspergillus niger and Aspergillus flavus strains showed no clear zone at 4 mg / disc.

The antimicrobial and antifungal activity of the white lotus leaf complex composition was higher than that of 70% ethanol extract, especially the Salmonella typhimurium, Pseudomonas aeruginosa, and Trichophyton mentagrophytes .

Inhibition zone (mm) 0.5mg / disc 1mg / disc 2mg / disc 4mg / disc Staphylococcus aureus
Bacillus subtilis
Bacillus cereus
Streptococcus mutans
Escherichia coli
Salmonella typhimurium
Pseudomonas aeruginosa
Vibrio parahaemolyticus
Aspergillus oryzae
Aspergillus niger
Aspergillus flavus
Trichophyton mentagrophytes -
-
-
-
+
+
-
-
-
-
-
- ++
-
++
+
++
++
++
++
++
-
-
+++ ++
-
+++
+++
+++
++
++
++
+++
-
-
++++ +++
+++
+++
++++
+++
+++
++
+++
++++
-
-
++++

Where + is less than 10 mm, ++ is 10 to 13 mm, +++ is 14 to 17 mm, and ++++ is 18 mm or more.

The antibacterial and antifungal activity of the six strains was measured by paper disc agar diffusion using a sample heat-treated at 100 ℃ for 30 minutes and at 121 ℃ for 15 minutes. Even after the heat treatment, the antimicrobial activity of the composite composition was almost unchanged and appeared to be stable to heat.

Therefore, when used as a preservative in most processed foods that require a heat treatment process to improve the palatability and shelf life of the food, it is expected to be easy to apply.

Inhibition zone (mm) 100 ℃, 30min 121 ℃, 15min Escherichia coli
Bacillus cereus
Vibrio parahaemolyticus
Streptococcus mutans
Aspergillus oryzae
Trichophyton mentagrophytes +++
+++
+++
++++
++++
++++ +++
+++
+++
++++
++++
++++

Where + is less than 10 mm, ++ is 10 to 13 mm, +++ is 14 to 17 mm, and ++++ is 18 mm or more.

The white lead leaf composite composition was adjusted to 0, 25, 50, 100, 200, 400, 800, 1,600 ppm (0, 0.025, 0.05, 0.1, 0.2, 0.4, 0.8, 1.6 mg / ml) 50 μl of each bacterial suspension was inoculated into 950 μl of the appropriate liquid medium mixture, and after 24 hours of incubation at a proper temperature, the absorbance was measured at 600 nm. 100 ppm of Staphylococcus aureus , 400 ppm of Bacillus subtilis, Bacillus cereus , Streptococcus mutans , Escherichia coli, Salmonella typhimurium were inhibited more than 50% at 200 ppm concentration, 63.80% at Pseudomonas aeruginosa, Vibrio parahaemolyticus at 75 ppm concentration. % Growth was inhibited and growth was completely inhibited in all 8 test bacteria at 1,600 ppm.

Inhibitory effect (%) Concentration (ppm) 25 50 100 200 400 800 1600 Staphylococcus aureus1
Bacillus subtilis
Bacillus cereus
Streptococcus mutans
Escherichia coli
Salmonella typhimurium
Pseudomonas aeruginosa
Vibrio parahaemolyticus 33.96
16.78
24.28
39.72
44.79
43.87
63.80
73.54 38.36
17.24
26.59
42.13
46.80
45.09
63.80
77.64 77.36
21.91
30.78
42.99
42.25
49.26
64.02
78.43 79.52
28.21
34.10
50.12
52.47
54.63
68.87
80.70 88.85
51.22
76.30
50.27
79.52
71.17
68.95
96.63 91.51
52.87
84.39
57.25
91.22
82.67
92.05
97.24 100
100
100
100
100
100
100
100

The white lead leaf composite composition was adjusted to 0, 25, 50, 100, 200, 400, 800, 1,600 ppm (0, 0.025, 0.05, 0.1, 0.2, 0.4, 0.8, 1.6 mg / ml) after incubation for 24 hours at the proper temperature by by 50 ㎕ inoculated to each of the spore suspension in the appropriate broth mixture 950 ㎕, as in the inhibition measures the minimum inhibitory concentrations of significant back lotus leaf hybrid composition results in Table 10 is determined by the naked eye Trichophyton Mentagrophytes 200 ppm, Aspergillus oryzae 800 ppm, Aspergillus niger and Aspergillus flavus showed minimal inhibition at 1,600 ppm.

Concentration (ppm) 25 50 100 200 400 800 1600 Aspergillus oryzae Aspergillus niger Aspergillus flavus Trichophyton mentagrophytes O
O
O
O O
O
O
O O
O
O
O O
O
O
X O
O
O
X X
O
O
X X
X
X
X

Where O is Inhibition and X is No inhibition.

Claims (2)

Antibacterial, characterized in that it comprises a white lotus leaf extract, a gall extract and a small fennel extract in a ratio of 2: 2: 2, and a white lotus leaf composition in which the plum extract is mixed in a ratio of 1: 4. And antifungal agents. 65 to 75% ethanol is added to the white lotus leaf to extract the white lotus leaf extract at 65 to 75 ℃ for 3 to 4 hours,
45 to 55% of ethanol is added to the gall to extract the gall bladder extract at 75 to 85 ° C. for 4 to 5 hours,
Extracting the small fennel extract at 75 to 85 ° C. for 4 to 5 hours by adding 45 to 55% of ethanol to the small fennel;
Extracting the plum extract by adding water to the plum and pressurizing for 2-3 hours;
Mixing the white lotus leaf extract, the gall bladder extract, and the small fennel extract in a ratio of 5: 2: 2 to form a white lotus leaf composition;
Antibacterial and antifungal production method comprising the step of mixing the white lotus leaf composition and the plum extract in a ratio of 1: 4 to create a white lotus leaf composite composition.

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