KR101270972B1 - Antifungal composition containing plant extracts - Google Patents

Abstract

The present invention relates to an antifungal composition comprising a plant extract, more specifically, a composition containing clove, cinnamon, licorice, cheongung, grapefruit seed extract and acetic acid and having excellent antifungal and antifungal effects.

Description

Antifungal composition containing plant extracts

The present invention relates to an antifungal composition comprising a plant extract, and more particularly to a composition having an excellent antifungal effect, characterized in that it contains clove, cinnamon, licorice, cheongung, grapefruit seed extract and acetic acid.

Persimmon is a deciduous arborescent of the persimmon tree distributed in central and northern China. Persimmon is one of the most nutritious fruits with less moisture than other fruits, but contains about 14% of glucose and fructose. Persimmon contains little organic acid but contains pectin and carotenoids. It also contains a lot of potassium and magnesium as tannins, vitamin C, vitamin A and inorganic salts that have a salty taste, prevent hypertension, and lower blood alcohol. Vitamin A is 400 units (IU) / 100g is very high and contains a lot of vitamin C. In particular, vitamin C in persimmon is a form of vitamin C precursor, and unlike ordinary vitamin C, it is not easily destroyed when exposed to heat, water, or air.

In addition, the tannin component of persimmon is good for digestive diseases such as gastric ulcer and diarrhea, due to the strong astringent action of tannic acid. Converging action of tannic acid is also effective for the recovery of burns. Persimmon tannins have long been used as a paralytic. A stroke is usually caused by high blood pressure, and refers to a paralysis state following cerebral hemorrhage. Gamma tannin is used as a stroke because it has a high blood pressure preventing effect. In addition, tannins are effective in preventing or treating frostbite or hand and feet, preventing bacterial infections and promoting the healing of inflammation and skin sores. In addition, tannins have a strong anti-toxic effect when applied to a wound when bitten by a snake. Tannin's most important effect is to bind nicotine and release it in vitro. Tannins also bind well with other harmful metals, detoxify, penetrate pathogens, bind to proteins and coagulate to kill bacteria.

In Dongbobom, he had to eat hongsi when he had sore throat or thirst. Persimmons and persimmon leaves are very beneficial to health because they are often taken as fruit, persimmon leaf tea, or porridge, which is useful for the prevention of circulatory diseases such as hypertension and adult diseases such as diabetes.

However, persimmon exhibits a number of functions that are beneficial to the human body as above, but the technology for long-term storage of persimmon is insufficient. Since ancient times, persimmons are eaten as it is, or semi-dry (dried persimmon), Hongsi, etc. There is a problem that bacteria or fungi easily breed. In the case of contaminated dried persimmons, the number of contaminants in the dried persimmons was about 1,000 times higher than that of normal dried persimmons. The main contaminants were blue mold> yeast> other bacteria. The most influential contaminant is known as blue mold. During the drying period, there is a risk of deterioration of quality due to incorporation of foreign substances, mold growth, and discoloration. Therefore, most farms are treated with sulfur fumigation to improve the merchandise of dried persimmons during drying. Sulfur fumigation treatment helps to improve the quality of dried persimmons, including anti-oxidation of tannins, prevention of dark brown discoloration of dried persimmons, inhibition of microbial propagation and drying promotion.

However, in the production of dried persimmon, sulfur fumigation treatment is known to have sulfur dioxide (sulfurous anhydride, SO ₂ ) adversely affecting the human body, and in particular, it is known to cause fatal harm to asthma patients. Is restricted to food additive legislation. However, in domestic dried persimmon production, most farms produce dried persimmon through sulfur fumigation, and thus the controversy of hazards caused by sulfur fumigation is constantly occurring.

Accordingly, it is an object of the present invention to provide a composition exhibiting antifungal activity for use in persimmon treatment in place of the sulfur fumigation method.

In addition, an object of the present invention is to increase the safety of the product by using a natural material harmless to the human body, to provide a dried persimmon improved antifungal power.

In order to achieve the above object, the present inventors tested the antifungal activity of various plant extracts in place of the sulfur fumigation method conventionally used in the production of dried persimmon, including clove, cinnamon, licorice, cheongung, grapefruit seed extract and acetic acid An antifungal composition was prepared.

The composition is clove extract 10-25% by weight, cinnamon extract 5-15% by weight, licorice extract 5-15% by weight, cheongung extract 1-10% by weight, grapefruit seed extract 1-10% by weight, acetic acid 45-60% by weight It is characterized by that.

The composition is characterized in that it is used for spraying or fumigation during drying of persimmons.

The composition is a clove extract, cinnamon extract, licorice extract, cheongung extract, grapefruit seed extract is dried clove, cinnamon, licorice, cheongung, grapefruit seeds, respectively, after grinding and grinding into powder and mixed with ethanol and water to extract It is obtained by filtration and concentration under reduced pressure.

The extract is characterized in that the mixed solvent of water and ethanol and each powder sample is mixed and extracted in a ratio of 4: 1, and then centrifuged to obtain a supernatant and then filtered with a filter paper.

Method for producing an antifungal composition of the present invention is as follows. Cloves, cinnamon, licorice, Cheongung, and grapefruit seeds are dried, ground and ground into powder. Each sample is shaken in a mixture of ethanol and water, shaken well, left under room temperature, filtered under reduced pressure and concentrated under reduced pressure. Obtain crude extract and use it as a sample.

Water and ethanol are mixed at a ratio of 1: 1 and each sample is mixed at a ratio of 4: 1 and extracted with shaking. The extract is centrifuged to obtain a supernatant, which is then filtered and used as each extract.

Clove extract 10-25 wt%, cinnamon extract 5-15 wt%, licorice extract 5-15 wt%, Cheongung extract 1-10 wt%, grapefruit seed extract 1-10 wt%, acetic acid 45-60 wt% Prepare an antifungal composition. When the clove extract is added more than 25% by weight, the unique flavor of the clove is too strong, the color of the persimmon changes to black as the dried persimmon progresses, and the persimmon and grapefruit seed extracts are added to the persimmon due to the peculiar bitter taste. Reduces the quality of. In addition, when the acetic acid is added more than 60% by weight, the sour smell peculiar to vinegar loses its fragrance during the drying period, but the acidity of the dried persimmon is changed.

The antifungal composition is immersed in the antifungal composition in the early stage of drying the persimmon or spraying the antifungal composition on the persimmon is harmless to the human body and the antifungal force is improved to facilitate the storage and storage of dried persimmon.

The composition prepared by the method of the present invention is harmless to the human body but inhibits the growth or reproduction of bacteria or fungi in dried persimmons.

In addition, dried persimmon prepared using the composition of the present invention is hygienic, uniform distribution of internal moisture and high sugar content. In addition, there is an advantage that the color and shape is maintained for a long time in the distribution process, the taste is better.

Figure 1 shows the morphological characteristics of the strain isolated from contaminated dried persimmon.
Figure 2 is a measure of the antifungal power of natural medicinal plant extracts against blue mold by the paper disk method.
Figure 3 is the antifungal force of the concentration of each extract for the fungus was measured by the paper disk method.
Figure 4 shows the change in color of dried persimmon after spraying clove extract, cinnamon extract, licorice extract, cheongung extract, grapefruit seed extract and acetic acid on dried persimmon, respectively.
5 is a measure of the antifungal power according to the ratio of the clove extract, cinnamon extract, licorice extract, cheongung extract, grapefruit seed extract and acetic acid by the paper disk method.
6 is a measure of the antifungal power according to the concentration of clove extract, cinnamon extract, licorice extract, cheongung extract, grapefruit seed extract and acetic acid by the paper disk method.
Figure 7 shows the change in the number of bacteria of dried persimmon sprayed test and control according to the storage period of each region.
Figure 8 shows the change in the number of molds of dried persimmon sprayed test and control according to the storage period of each region.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these Examples are only illustrative description of the present invention and the scope of the present invention is not limited to these Examples.

Example  1: Preparation of extracts of natural medicinal plants

1-1. material

The natural medicinal plants used in the experiment were purchased from Geumsan Medicinal Center and Seoul Kyungdong Herbal Medicine Market. The scientific names and scientific names of these collected samples are shown in Table 1. The collected medicinal plants are dried in a drier at 50 ° C. for 2 days, and then ground to a powder. 20 g of each powder sample is taken into a 500 ml Erlenmeyer flask, mixed with ethanol and water, shaken well and left under room temperature in the dark. It was. After 5 days under reduced pressure filtration and concentrated under reduced pressure at 40 ℃ with a rotary vacuum concentrator (N-1001S-WD) to obtain a crude ethanol extract was used as a sample.

1-2. Extract Preparation Method

Extraction and yield measurement of 150 medicinal plants were carried out as follows. Water and 99% ethanol were prepared at a ratio of 1: 1, and then the ratio of the solvent and the sample was mixed at 4: 1 and extracted with shaking at 30 ° C. at 200 rpm for 12 hours. The extract was centrifuged at 16,000 x g for 10 minutes to obtain a supernatant, which was then filtered through filter paper (Whatman No. 2).

Experimental Example  1: Isolation of contaminated dried persimmon

The dried persimmons were pulverized with a blender (HR-2094) to separate the bacteria from the normal and contaminated dried persimmons using 0.85% NaCl. That is, 0.1 ml of the diluted solution was smeared into each medium and incubated in a 25 ° C. incubator (DS-BIP250) for 3 days. At this time, LB, MRS, NB, YM, YPD, Czapek-Dox and PDA were used as the culture medium. On the other hand, after culturing, the morphological characteristics such as shape, size, color selection, and gloss of colonies appeared in each medium were observed and confirmed, and each strains expected to be different strains were tooth-picking. In order to screen the fungus while screening the fungus while inhibiting the growth of bacteria from the dried persimmon raw materials, the experiment was divided into antibiotic-added medium and no-added medium, and the antibiotic used was streptomycin sulfate salt (Sigma, S6501). ) Was added to 50 μg per ml of medium.

The percentage of contaminants isolated from dried persimmons was the highest in normal dried persimmons, ranging from 3.09 × 10 ² to 6.18 × 10 ³ CFU / g, depending on the type of medium. Next, yeasts ranged from 3.00 × 10 ² to 3.45 × 10 ³ and other types of bacteria from 1.80 × 10 ² to 3.09 × 10 ³ . Therefore, the main contaminants of normal persimmons were blue mold>yeast> other bacteria.

In the case of contaminated dried persimmons, one species of blue mold was the main contaminant, followed by light purple and white yeast. For light purple yeast, 2.41 × 10 ⁵ ～ 3.8 × 10 ⁶ CFU / g, White yeast ranged from 1.00 × 10 ⁵ to 3.8 × 10 ⁶ CFU / g.

      From the results, the main contaminants of contaminated persimmons were blue mold> two types of yeast (light purple and white), and the most contaminants which had the greatest effect on the quality of dried persimmon during drying or storage of dried persimmon were blue mold. You can conclude.

      In this experiment, after collecting different bacteria among the isolated strains of each medium, K2-1, K-1, K-3 for mold, H2-1, H2-2 for yeast, and S-1 for bacteria , S-2, S-3, S-4, S-5, S-6, SC-1, S2-1, S2-2 and S2-3, respectively, were numbered and the results are shown in FIG.

       As shown in FIG. 1, in the case of mold, K2-1 was estimated to be a blue mold, and in the case of yeast, H2-1 was estimated to be an ivory yeast and H2-2 was a light purple yeast. In bacteria, about 10 strains were isolated, S-3 is white wrinkles, S-4 is Bacillus strain, S-5 is Bacillus strain, similar to S-4. The shape is similar to mold. In particular, S2-2 was analyzed to have a yellowish shine unlike other bacteria, and S2-3 and SC-1 were found to be white as fungi-like bacteria.

Experimental Example  2 Identification of Strains Isolated from Contaminated Dried Persimmon

Among the strains isolated from contaminated dried persimmon, the major strains of dried persimmon were identified by 16SrDNA nucleotide sequence separation and 16S ribosomal DNA sequencing method.

Table 2 shows the results of the identification of strains isolated from contaminated dried persimmons. For yeast and fungal strains, the nucleotide sequences of the 26S D1 / D2 and ITS regions of each strain were examined and compared with the nucleotide sequences of the strains registered in NCB GenBank (Yeast strain) / CBS DB (fungal strain). , H2-2 Citeromyces matritensis , Metschnikowia sp.

Molds K-1, K2-1 and K-3 are Cladosporium sp., Penicillium sp. And Aspergillus sp.

In the case of bacteria, 16S rRNA gene sequences of each strain were examined and compared with those of strains registered using NCBI GenBank and EzTaxon server 2.1. S-1 and S2-2 were Acinetobacter , respectively. sp. And Microbacterium sp.

Experimental Example  3: Antibacterial and Antifungal Activity of Natural Medicinal Plant Extracts

Dried persimmon contaminants (K2-1, K-3, K-1, S-1, S-2, S-3, S-4, S-5, The antimicrobial and antifungal properties of S-6, S2-1, S2-2, S2-3, SC-1, H2-1, H2-2) were analyzed by the paper disk method, and the results are shown in Table 3.

(Unit: mm)

Among the 150 natural medicinal plant extracts selected in this experiment, 47 extracts with antimicrobial activity against 15 bacteria isolated from contaminated dried persimmons were analyzed. However, in the case of natural extracts with antimicrobial activity, 6 to 7 natural products with comprehensive activity against isolated bacteria were analyzed.

In general, about 47 extracts with antimicrobial activity were found to have excellent activity against bacteria, and natural extracts showing antifungal activity against blue fungus, the main contaminant of dried persimmon, were analyzed as extracts of clove, licorice, celery, and cinnamon. .

Persimmon leaves showed good activity against main contaminants of dried persimmon, but were analyzed as pesticides and excluded from this experiment. Natural extracts with the best antibacterial and antifungal activity were analyzed as clove extract. In the case of clove extract, as shown in Table 2, the clear zones were 20 mm, 24 mm, and 51 mm for the K2-1, K-3, and K-1 bacteria, and the antifungal activity against the fungus was very excellent. In particular, the extract of clove has a wide range of antifungal and antibacterial activity against the H2-1, H2-2 group and bacteria, etc., was expected to be widely used in controlling persimmon pollutants.

Experimental Example  4: blue mold ( Penicillium sp For.) Antifungal activity

The antifungal activity of blue mold, which is the main contaminant of dried persimmon, of the extracts of 150 kinds of natural medicinal plants used in this experiment was analyzed by the paper disk method, and the results are shown in FIG. 2.

As shown in Figure 2 extracts of clove, cinnamon, cheongung, licorice were analyzed to have excellent antifungal activity against blue fungus. That is, the clear zone of the clove extract and the licorice extract was 12 mm, the cinnamon extract 10 mm, and the uterine extract 9 mm with respect to 50 µl of these extracts.

The composition of the present invention was selected from the extract of clove, cinnamon, licorice, cheongung out of 150 kinds of medicinal plants through the results of Experimental Example 3, Experimental Example 4, was configured by further selecting acetic acid, grapefruit seed extract.

Experimental Example  5: blue mold ( Penicillium sp By concentration of each extract for.) Antifungal activity

Blue fungus ( Penicillium) , the main contaminant of dried persimmon In order to verify the antifungal activity according to the concentrations of clove extract, cinnamon extract, celery extract, and licorice extract, which showed antifungal activity in sp. The results are shown in Table 4 and FIG. At this time, as a control for each extract was used nystatin (Nystatin) and mycoazole (Mycoazole).

The antifungal activity of each extract formed a clear zone with an average of 8-16 mm, and in particular, the cinnamon extract formed a clear zone at a low concentration of 10 μl. Licorice extract and cheongung extract were analyzed to show antifungal activity from about 30 μl concentration.

When loading 30 μl of clove extract (50% ethanol extract), 50 μl of stock solution of Nystatin (1.5 μg / mL) and Mycoazole (10 μg / mL), respectively, used as a control. High antifungal activity was seen similarly to when loading.

Experimental Example  6: extract In dipping  Dried persimmon chromaticity

In order to analyze the effect of each extract on the color and aroma of dried persimmon, the selected persimmons, cinnamon, licorice, cheongung, grapefruit seed extract and acetic acid were sprayed on dried persimmons, respectively. The change is shown in FIG. 4.

As shown in FIG. 4, the clove extract showed superior antifungal activity compared to other extracts, but as the drying of the dried persimmon proceeded, the color of the dried persimmon was changed to black. In addition, cloves, like cinnamon, are very strong in clove, which has a strong influence on the quality of dried persimmon.

In the case of licorice, the licorice-specific aroma was less rejected, not dark, and did not significantly affect the color of dried persimmon. On the other hand, the cheongung had no significant effect on the appearance of dried persimmons, but the fragrance was poor and the antifungal activity was relatively lower than that of other extracts.

In addition, the grapefruit seed extract used did not significantly affect the color and aroma of the dried persimmon, but it was shown that the bitter taste peculiar to the grapefruit seed extract may cause deterioration of the dried persimmon. Apple cider vinegar has a sour smell that is unique to vinegar, but the fragrance is lost during the drying period, so there is no problem in terms of smell.

Experimental Example  7: setting the mixing ratio of the extract

After setting the mixing ratio of the extract of clove, cinnamon, licorice, cheonggung and acetic acid, grapefruit seed selected in the present invention and then using it to analyze the antifungal force against the fungus ( Penicillium sp.) The results are shown in Table 5 and Figure 5 is shown.

The clear zone size was 10-14 mm depending on the total 28 mixing conditions. The blending ratio of the clove extract and cinnamon extract was found to be the highest when the ratio is 20-30% of the total blending ratio.

Among the six extracts, the antifungal activity was lower than that of the other vinegar when the ratio of the other extract except the vinegar was 10-40% or more.

That is, the total blending ratio of vinegar was the best antifungal power when the ratio of the other extract is about 10-30%. In the case of vinegar, the antifungal activity was remarkably excellent when the vinegar accounted for more than 50% of the total meal ratio.

The concentration of each extract was 50 μl of cinnamon extract, 30-40 μl of clove extract, 10% acidity of vinegar, and 20 times of grapefruit seed extract. Controls of Myconazole and Nystatin showed 12 mm clear zones at 1.5 μl and 10 μl concentrations of 1 mg / ml, respectively. Potassium metabisulfite showed a clear zone of 12 mm at 10,000 ppm. Shown (FIG. 6).

Example  2: preparation of antifungal composition

The present inventors prepared an antifungal composition by combining 18.18% by weight of clove extract, 9.09% by weight cinnamon extract, 9.09% by weight licorice extract, 4.55% by weight celery extract, 4.55% by weight grapefruit seed extract, 54.54% by acetic acid.

Control

Stock solutions of Nystatin (1.5 μg / ml) and Mycoazole (10 μg / ml) were sprayed onto 50 μl dried persimmon, respectively.

Experimental Example  8: Changes in the Number of Bacteria and Molds of Dried Persimmon During Drying

In Experimental Example 8, in order to prove the efficacy of the composition of Example 2, Sangbuk Campus (386 Sangji-dong, Sangju-si), Cape Persimmon Distribution Center (214 Deodong-dong, Sangju-si), Changju 1-ri, Sori 1-ri, 21, Sojang 1-ri, Cheongha-myeon, Cheongha-myeon, 319, etc. In order to ensure accuracy by selecting four places, the dried persimmon drying system was manufactured by hand, and the control and test zones were sprayed at an average spraying rate of 300 ml for each of the four regions (November 5, 2010, 11, 2010). On April 6, 2010, November 8, 2010, November 11, 2010, a total of four sprays were installed), and 600 stripping strips were installed in each drying system in each region. By January 7, experiments were conducted on bacterial and mold number changes and are shown in FIGS. 7 and 8.

Referring to Figures 7 and 8, the number of bacteria and fungus in the test zones showed less values in all four regions than the control, it was confirmed that the composition prepared by the method of the present invention has the effect of antibacterial and antifungal.

.

Claims (5)

Clove Extract 10-25%, Cinnamon Extract 5-15%, Licorice Extract 5-15%, Cheongung Extract 1-10%, Grapefruit Seed Extract 1-10% and Acetate 45-60% Antifungal composition for drying persimmons.
delete The method according to claim 1,
The composition is dried antifungal composition for drying persimmon, characterized in that it is used for spraying or fumigation during drying of persimmon.
The method according to claim 1,
The composition is a clove extract, cinnamon extract, licorice extract, cheongung extract, grapefruit seed extract is dried clove, cinnamon, licorice, cheongung, grapefruit seeds, respectively, after grinding and grinding into powder and mixed with ethanol and water to extract Antifungal composition for drying persimmon obtained by filtration and concentration under reduced pressure.
The method according to claim 1,
The clove extract, cinnamon extract, licorice extract, cheongung extract, grapefruit seed extract is a mixture of water and ethanol and extract each powder sample of clove, cinnamon, licorice, cheongung, grapefruit seeds in a weight ratio of 4: 1, The antifungal composition for persimmon preservation, which is obtained by centrifugation and taking the supernatant, followed by filtration with filter paper.

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