KR20010045830A - The antibacterial and antioxidant composition of natural products - Google Patents

Abstract

PURPOSE: A crude drug composition having optimum antibacterial and antioxidation action through the screening, selection and extraction of natural extracts having antibacterial and antioxidation function alone or mixtures thereof is provided, which has safety to human body and can be used for cosmetics and goods in a kitchen. CONSTITUTION: The crude drug composition consists of 0.5 to 5% by weight of a barberry root and Rubus coreanus Miq., 0.05 to 10% by weight of Prunus mume S. et Z. and gallnut, 0.1 to 5% by weight of clove respectively. The composition exhibits antibacterial effect against harmful microorganism.

Description

The antibacterial and antioxidant composition of natural products

The present invention relates to herbal extracts having antibacterial and antioxidant functions and compositions thereof. More specifically, the present invention is applied to each of the natural extracts present in nature by applying heat to the aqueous extract to measure the antimicrobial function and antioxidant capacity for each extract and antibacterial and antioxidant that can be universally applied through the appropriate combination of these The present invention relates to a method for producing a natural composition having a function.

Natural antibacterial extract is obtained by cutting off the water, so it is resistant to heat, so it is easy to process and does not lose its antimicrobial activity by heat, and above all, it is used for conventional antimicrobial chemicals that are resistant to various antibiotics and pesticides. Since it exhibits antimicrobial activity by different activity mechanisms (mechanisms), there is an advantage that there is little possibility of causing resistance. In addition, because it is a natural extract has the advantage that the toxicity of the substance itself is extremely weak than by the general chemical synthesis. Therefore, natural antibacterial extract has a great potential for industrial application in pharmaceutical, food and agriculture fields.

Existing antimicrobial agents are mostly manufactured through chemical synthesis, and their use ranges from agricultural pesticides developed for agricultural use to antibiotics used in medicine. However, as their use increases, the emergence of resistant bacteria is frequent, and recently, the situation is serious enough to anticipate the outbreak of super bacteria in the near future.

Recently, methods for mass production of antimicrobial peptides found in the skin of microorganisms or higher animals have been developed, but they are relatively weak to heat because of their physicochemical properties, and are generally used because they are very susceptible to protease, which is widely found in nature. There is a limit to this. Moreover, they lack the selectivity of bacteria, fungi and animal cells due to their mechanism of action, which makes them difficult to use as pharmaceuticals. It is relatively expensive. Therefore, there are many problems to be solved until they are utilized in agriculture and general fields.

The necessity of developing a non-toxic and pollution-free antimicrobial agent derived from nature has been raised in recent years. Food preservatives derived from various chemical synthesis that have been used for a long time are now being used even in the food field. In addition, in the field of agriculture, a movement that uses organic fertilizers a lot in the name of environmentally friendly agriculture has been developed recently, and there is even a claim to use natural enemies instead of pesticides that destroy the environment around the world. Therefore, the only solution that can solve all these problems is to find out the potential functions of various natural products in the natural world and to utilize their unique functions in our lives.

On the other hand, in Korea, there are currently no methods for combating various disease bacteria, which are difficulties in the cultivation process, such as the problem of cultivating soybean sprouts using pesticides regularly appearing in the broadcasting media, or the mushroom cultivation of special crops. The latter reason is that it is not easy to develop drugs that selectively exterminate only bacteria other than mushroom because mushroom itself is included in taxonomic fungus. Another problem is the profitability problem of specialized pesticide development companies. Therefore, the natural antibacterial extract has the advantages of very low development cost, environmentally friendly and no pollution and toxicity problems.

The present invention is a natural extract of natural extracts having excellent antimicrobial activity selected through screening of antibacterial and antioxidant functions of natural extracts alone or of an organic, nontoxic natural extract having an optimal antimicrobial and antioxidant activity through the proper composition ratio thereof. Provided are methods for preparing the composition.

Figure 1a is a table evaluating the antimicrobial activity against various microorganisms of the antimicrobial active extract in one embodiment of the present invention.

Figure 1b is a table measuring the antioxidant activity of the natural herbal extract in one embodiment of the present invention.

Figure 2a is a table measuring the antimicrobial effect effective concentration of the natural extract with antimicrobial activity in one embodiment of the present invention.

Figure 2b is a table showing the antimicrobial activity of the natural antimicrobial composition in one embodiment of the present invention.

The present invention relates to a method for preparing a herbal extract composition having a natural antibacterial and antioxidant function. The natural extraction composition of the present invention is a water-extracted through the heating of various herbal medicines in nature and search for their antibacterial and antioxidant functions, respectively, to broaden the broad antimicrobial (broad-spectrum) against various harmful microorganisms (broad-spectrum) The present invention relates to an antimicrobial natural aqueous extract and a method for preparing the composition thereof, which are prepared by appropriate mixing of extracts having respective activities.

Hereinafter, the present invention will be described in detail with reference to Examples. These examples are only to specifically describe the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples. In particular, the natural extracts of the specific antimicrobial and antioxidant functions exemplified in the embodiments of the present invention are only preferred specific examples, and the natural antimicrobial agents exhibit a relatively broad antimicrobial activity through the respective antimicrobial and antioxidant natural extracts or appropriate mixtures thereof. The method for producing the composition falls within the scope of the present invention. In particular, the antioxidant function discussed in the present invention is not directly related to the antimicrobial function, and for the purpose of constructing the claims of the present invention, the antioxidant function of the herbal medicine was further investigated and applied to the food field.

Example 1 Preparation of Herbal Extracts and First Search of Antimicrobial Function

Various herbal medicines used in this example were selected through literature studies introduced in various herbal medicine data collections while being easy to purchase in Korea. The details are shown in the following table.

Medicinal name used for search

50g of each medicine was prepared and extracted twice with 500ml of water for about 3 hours using a commercially available domestic electric bath. After extraction, 2 to 5 ml volume of each herbal medicine was taken to remove water for 12 to 24 hours in a reduced pressure dryer, and then dried extract of each herbal medicine was obtained. 1 ml of distilled water was added to a certain amount of dried matter of each medicinal herb, and the concentration of each medicinal extract was arbitrarily configured. The unit concentration of each medicine is shown in the following table.

Unit concentration (mg / ml) of each herbal extract prepared

After preparing the natural extract sample obtained above, 100 microliters of a specific microbial culture solution was added and solid agar medium (2% glucose, 1% peptone, 0.5% yeast extract, 2% bactoagar) was dissolved in water and sterilized by autoclave. After placing the antibiotic test discs (antibiotics disc assay, PGC scientifics) on the solidified medium), 30 microliters of specific extracts were added to each disc. The antibiotic test discs used in this antimicrobial evaluation test were standard sizes (0.95 cm diameter, 3/8 inch) recommended by the US FDA and WHO.

In this example, the microorganisms used for the evaluation of the antimicrobial for the primary search purposes were conducted on representative strains related to various disease occurrences (Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, Baker's yeast), and a sample was put on each disk. Then, after incubating for 12 to 24 hours or more at the optimum culture temperature of a given bacterium, the antimicrobial activity was determined based on the size of the plaques formed around the disc (areas in which cell growth was suppressed or cells were broken up and formed transparent) It was. In each antimicrobial test, the same amount of microbial growth inhibitor (Gibco-BRL) consisting of 100 U / ml Penicillin G, 250 ng / ml Streptomycin sulfate, and 0.0085% Amphotericin B was used as a control. As a result of the antibacterial test obtained through this process, six kinds of herbal medicines showing superior effects to the control group were selected. These were five plums, five pears, cloves, bokbunja, rhubarb, and sorrel.

Example 2. Investigation of Antimicrobial Activity of Selected Natural Aqueous Extracts

In order to further search the antimicrobial activity of the six herbal extracts retrieved in Example 1, the search was expanded to various microbial species. The concentration of each extract used in the evaluation in the present Example was maemae (31mg / ml), baejaja (24mg / ml), joiner (11mg / ml), rhubarb (17mg / ml), cloves (14mg / ml), bokbunja (12.3 mg / ml or 32 mg / ml to indicate the treatment concentration on the figure). The irradiation method was carried out in the same manner as in Example 1 and the results of the present example are shown in Figure 1a. In Figure 1a, the symbols in the table are + (equivalent or weaker than the control), ++ (usually effective), +++ (excellent), ++++ (excellent), +++++ (very excellent). Marked as. -Indicates that the effect was not significant. The antimicrobial test results showed that the six natural herbal extracts showed extremely limited antimicrobial activity against fungi such as yeast, while showing excellent antimicrobial activity against toxic or normal bacteria. The microorganisms used in this example were carried out by limiting the range to aerobic microorganism types.

Example 3. Determination of Antioxidant Degree of Natural Aqueous Extract

Antioxidant levels of the herbal extracts used in Examples 1 and 2 were measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH) (Kor. J. Pharmacogn. 24 (4) 299-303 (1993)). It was. Vitamin C was used as the standard for the antioxidant test. Specific methods are as follows. Each natural extract retrieved in Example 1 was diluted with 100% methanol, with dilution ranges of 1 mg / ml, 0.5 mg / ml 0.1 mg / ml 0.05 mg / ml, 0.01 mg / ml 0.005 mg Dilute appropriately to / ml, 0.001mg / ml. Standards are prepared in a similar concentration range, then the DPPH reagent is dissolved in 100% methanol to a final concentration of 0.2 mM. Antioxidant measurement was performed using a 96-well plate, in which 200 microliters of DPPH solution was injected into each ball, and then 50 microliters of the diluted extract was injected with two repeats of each ball. . In order to exclude the effect of the basic absorbance of the sample for accurate test, only the sample is added to the empty hole next to the same condition.These balls are added with 100% methanol, not DPPH solution. Genie subtracted the specific wavelength absorption capacity after the test was completed. In this way, measurements at specific wavelengths are only influenced by the results of the antioxidant reaction. After the solution was mixed and left at room temperature for 30 minutes, the antioxidant capacity was measured at a wavelength of 517 nm in a 96-hole plate measuring instrument (EIA reader). Assuming that the absorbance of the DPPH solution without the sample was 100, the ability to cancel this measurement by 50%, i.e., the amount of extract required to obtain the RC50 value, was shown in the table in FIG. In the table of FIG. 1B, the lower the value of the RC50 value, the higher the antioxidant degree. Antioxidant measurement results showed that the five folds showed much higher antioxidant levels than the standard. Therefore, it was confirmed that the gall bladder possesses high antioxidant function and excellent antibacterial function.

Example 4 Determination of the Range of Antimicrobial Activity of Natural Extracts

Among the herbal extracts obtained in Example 1 were selected rhubarb, ume, bokbunja, cloves, gallengja and the like, and the extracts are stored at room temperature to remove the precipitate formed by centrifugation. The supernatant liquid from which the precipitate was removed was dried under reduced pressure to obtain a dried product, and the weight thereof was prepared to have a final concentration of 0.01%, 0.05%, 0.1%, 0.5%, and the like. At this time, the dilution of the dried liquid is prepared a new liquid medium except agar in the solid agar medium prepared in Example 1 and used as a diluent. The prepared solution will later be used as a culture for a specific bacterium, so the dilution of extract prepared at each concentration is passed through a 0.2 micrometer (um) sterile filter to eliminate the possibility of contamination of all microorganisms that may be derived from the medium and the sample. 50 microliters or 100 microliters of test microorganisms pre-cultivated in the media solution of each extract concentration were added to each medium solution by incubation for 5 to 7 hours in the incubator, and then the relative By comparing and measuring the number, it is possible to confirm the effect of inhibiting the growth of microorganisms by concentration according to each extract. The growth of microorganisms in the medium can be determined by incubating each microorganism at an optimal temperature for a given time after inoculation and measuring the absorbance at 600 nm. If the growth of microorganisms is good, the absorbance is high and growth is inhibited. The value of absorbance becomes very low. Relative comparisons of these numbers can indirectly confirm the extent of growth. The measurement result of this Example is shown in FIG. 2A. As a result of the test in this example, it can be seen that the lowest growth inhibition concentration of E. coli (E. coli) is 0.01%, 0.5% for clove and bokbunja, respectively, 0.01% for the rhubarb. In this way, the ermine was extensively investigated for various intestinal harmful bacteria, and the minimum action concentration was 0.01-0.05% for each bacterium. On the other hand, the five gall extracts were excluded from the experiment by inducing precipitation by reacting with certain components in the medium when mixed with the medium. However, as shown in FIG. 1a, the excellent antimicrobial activity of the five oocytes detected in the solid agar medium was confirmed in the liquid medium experiment. If there was no problem of precipitation, it was assumed that the liquid medium measurement showed higher antimicrobial activity.

Example 5. Evaluation of antimicrobial activity through complex composition of antimicrobial natural extracts

By properly mixing the respective natural extracts based on the results obtained in Example 4, it is possible to construct an optimal natural extract composition showing antimicrobial activity against a variety of microorganisms. For example, in Figure 1a, the hawk showed antimicrobial activity against E. coli, but there was no activity in the case of the gall bladder. However, in the case of gall bladder, ume showed the opposite activity against the bacterium without antimicrobial activity. Therefore, in this example, the antimicrobial activity of each natural extract composition was evaluated for representative strains (performed in Example 1) according to the ratio of each composition. The test was carried out on the solid agar medium with the same disk assay as in Example 1 and the evaluation scale applied the criteria as in Example 2 and the results are tabulated in FIG. 2B.

The extract or composition having antibacterial and antioxidant function prepared according to the present invention is a non-toxic natural functional composition that is non-toxic to human body, which has a high possibility of application as an antimicrobial purpose in crop cultivation field and an antimicrobial preservative in food field and also has antimicrobial function. It can be used as a new material in the field of applicability to cosmetics and kitchen utensils and pharmaceuticals. In particular, the results in Example 5 show that the proper combination of each antimicrobial herbal extracts are complementary in the antimicrobial properties of each extract. Therefore, it can be seen that the herbal extract composition of the present invention exhibits general antimicrobial activity against harmful microorganisms.

Claims (4)

Natural composition with complementary antibacterial and antioxidant functions derived from herbal extracts The method of claim 1, wherein the antimicrobial herb extract is a method of producing a natural composition consisting of rhubarb, lumberjack, ume, locust, bokbunja, cloves in an independent or suitable mixing ratio. The method according to claim 2, wherein the anti-oxidant material comprises fivefold, cloves, bokbunja and joiner, independently or at least two or more kinds thereof are mixed. The method according to claim 2, wherein the concentration of each of the herbal extracts constituting the composition having an antimicrobial function is in the range of 0.5% to 5% of the concentration of sulfur and bokbunja and at least 0.05 Complex natural herbal composition that is included in the range of up to 5 ~ 10% in the range of% ~ 0.1% and clove is included in the range of 0.1% ~ 5%, and optimally mixed with two or more herbal extracts, such as ume, galleng Manufacturing method.