KR20190079875A - An anti-microbial compositions comprising Abeliophyllum distichum extract as an active ingredient - Google Patents

Abstract

The present invention relates to an antimicrobial composition containing an extract of Abeliophyllum distichum as an active component, which has excellent antimicrobial activity against harmful strains, scavenging ability such as DPPH and ABTS radicals, and inhibitory activity of active oxygen. For the same, the present invention contains the extract, extracted from any one or two or more sites selected among leaves, stems, fruits, roots, bark, and seeds of Abeliophyllum distichum, as the active component.

Description

An antimicrobial composition comprising an extract of Alaska pollack as an active ingredient (An anti-microbial composition comprising Abelophyllum distichum extract as an active ingredient)

The present invention relates to an antimicrobial composition comprising an extract of Helicobacter pylori, which has excellent antimicrobial activity against harmful strains, scavenging activity such as DPPH and ABTS radicals, and active oxygen inhibition as an active ingredient.

Antibiotics are generally referred to as antimicrobial agents, and particularly antimicrobial agents against bacteria, specifically, bacteria that inhibit systems for synthesizing cell walls and proteins, Or made from such materials.

Antibiotics are mainly extracted from molds, and today they are used to treat diseases caused by bacterial infections. The most common antibiotic is penicillin, made in 1928 by the British doctor Alexander Fleming.

Penicillin was the first antibiotic manufactured by mankind to respond to bacteria in earnest. A representative antibiotic developed after penicillin is methicillin, which is considered to be more effective than phenylcylin. Imatinocin was produced by partially modifying the chemical structure of penicillin.

The methicillin is classified as a beta-lactam class of antibiotics. The beta-lactam antibiotic binds to penicillin-binding proteins (PBPs) By removing the activity, it is effective.

On the other hand, antibiotic-resistant bacteria refer to bacteria that are resistant to certain antibiotics and do not listen to their effects. This includes, for example, penicillin resistant Staphylococcus aureus , which does not heal the above-mentioned penicillin efficacy. In addition, methicillin-resistant Staphylococcus aureus (MRSA) has been reported to academia for the first time in 1961 and has since become a major pathogen in hospitals worldwide. The MRSA has been found to have a unique gene that is resistant to penicillin or methicillin antibiotics. It is reported that MRSA is not infectious to healthy people, and infects mainly patients with weak immunity or patients who have undergone surgery, but it causes death due to sepsis or pneumonia when infected.

As the infections caused by pathogenic microorganisms resistant to the existing chemical-derived antibiotics are increased, recently, antimicrobial agents derived from existing chemical substances have become problematic in terms of side effects and resistance, and the demand for development of antimicrobial substances derived from natural materials has increased As a result, attempts have been made to develop various medicines or antibiotics using natural products.

Accordingly, the inventors of the present invention have made intensive studies on a novel substance which comprehensively considers functionality such as stability, economical efficiency and antibacterial activity to human body as a result of such an attempt. As a result, Activity and the like, while confirming that it is harmless to the human body through experiments. Thus, the present invention has been completed.

Korean Patent Laid-Open No. 10-2017-0122315 (June 10, 2017) Korean Patent Publication No. 10-2013-0074172 (Apr. Korean Patent No. 10-0706131 (April 04, 2007)

DISCLOSURE OF THE INVENTION An object of the present invention is to provide an antimicrobial composition comprising an extract of Helicobacter pylori, which has excellent antimicrobial activity against DPPH and ABTS radical scavenging ability and active oxygen, .

In order to achieve the above object, the composition of the present invention is characterized by comprising, as an active ingredient, an extract extracted from any one or two or more parts selected from the leaves, stem, fruit, root, do.

In addition, the active ingredient may be characterized in that it contains either or both of the components of verbascoside or polyphenol.

On the other hand, the extract may be prepared by pulverizing any one or two or more sites selected from leaves, stems, fruits, roots, husks and seeds of the brackish tree to prepare a pulverized material, and adding water or ethanol ) Is mixed and allowed to stand at room temperature for 1 to 2 days, or the extract is produced under heating, reduced pressure and vacuum.

The extract may also contain at least _{one selected from the group} consisting of purified water, C ₁ -C ₄ alcohols, glycerin, acetone, hexane, ethyl acetate, butylene glycol, propylene glycol ), Dichloromethane, chloroform and ethyl ether, or a solvent extraction or fractionation using a mixture of two or more of them.

On the other hand, the composition is characterized in that it is in the form of a pharmaceutical composition for treatment, or in the form of a health functional food, or in the form of a cosmetic composition, or in the form of a food additive or a feed additive.

Also, the composition is added to at least one selected from the group consisting of medicines for the prevention of food poisoning, foods for preventing food poisoning, fibers, daily necessities, building materials requiring antibacterial activity, and households.

The antimicrobial composition of the present invention has an excellent antimicrobial activity against harmful strains, DPPH and ABTS radical scavenging ability, and active oxygen inhibition.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a photograph showing antimicrobial activity of extract extracts and alcohol extracts of Streptococcus mutans against the leaf extract of Helicobacter pylori according to a preferred embodiment of the present invention. FIG.
2 is a photograph showing the antimicrobial activity of the extracted solvent and the butylene glycol extract on the Candida albicans of the Leucocephala leaf extract according to a preferred embodiment of the present invention.
FIG. 3 is a photograph showing the antimicrobial activity of extracts of Solanum tuberosum L. and extracts of Staphylococcus aureus against the extract of Solanaceae according to a preferred embodiment of the present invention.
FIG. 4 is a photograph showing antimicrobial activities of E. coli and Salmonella typhimurium extract of Solanum tuberculosis leaf extract according to a preferred embodiment of the present invention, respectively. FIG.
FIG. 5 is a graph showing the DPPH radical scavenging activity of Leucocephala leaf extract according to a preferred embodiment of the present invention.
FIG. 6 is a graph showing the ABTS radical scavenging activity of Leucocephala leaf extract according to a preferred embodiment of the present invention.
FIG. 7 is a graph showing the FRAP performance of Leucumber leaf extract according to a preferred embodiment of the present invention.
FIG. 8 is a graph showing the reducing power (reducing power) of the extract of Leucocephala leaves according to a preferred embodiment of the present invention.
9 is a graph showing verbascoside standard curve and spectral analysis according to the analysis conditions of optimal HPLC of Helicobida plant extract according to a preferred embodiment of the present invention.

The antimicrobial composition according to the present invention can be used as an effective ingredient in extracts from any one or two or more sites selected from leaf, stem, fruit, root, And may contain any one or both of the pharmacologically acceptable transition enzymes or excipients.

A method for extracting the extract of Helicobacter pylori according to a preferred embodiment of the present invention is a method for grinding a leaf, a stem, a fruit, a root, a skin, And the mixture is mixed with water or any one of ethyl alcohol and ethanol and allowed to stand at room temperature for 1 to 2 days. Alternatively, the extract can be prepared under heating, reduced pressure and vacuum.

In a further embodiment, the extract is selected from the group consisting of purified water, C ₁ -C ₄ alcohols, glycerin, acetone, hexane, ethyl acetate, butylene glycol, The solvent may be prepared by solvent extraction or fractionation using a mixture of any one or more of propylene glycol, dichloromethane, chloroform and ethyl ether.

Meanwhile, as an embodiment for extracting the extract of Myristella japonica according to the present invention through solvent extraction, first, the desired part is cut, washed, dried and crushed. At this time, washing and drying and crushing of the tree species (including leaves, stems, fruits, roots, husks and seeds) is to increase the contact surface with the solvent.

The Abelophyllum distichum , Nakai) is a plant of the ash tree, and it is called "the fine line tree" which resembles the round fan of the fruit. This line is a shrub that lives only in Korea and is the world's only natural monument of one genus, It has been verified through various studies that it has a large amount of polyphenol component of 10 times.

In particular, verbascoside, which is an active ingredient of Spirulina extract, is a compound belonging to phenylpropanoid glycoside, and various physiological activities such as antioxidative, anti-inflammatory, antibacterial and immunosuppressive effects have been reported. (Wu et al., 2007; Cardinali et al., 2012).

On the other hand, the extracted solution can be prepared by extracting the solution containing the active ingredient through the solvent extraction method for the pulverized tree species (including leaves, stems, fruits, roots, husks and seeds).

Herein, the active ingredient may include one or both of the components in verbascoside or polyphenol. In one embodiment for extracting the active ingredient, the solvent used in the solvent extraction method Examples of the solvent include purified water, C ₁ -C ₄ alcohol, glycerin, acetone, hexane, ethyl acetate, butylene glycol, propylene glycol, dichloromethane dichloromethane, chloroform and ethyl ether, or a mixture of two or more of them may be used.

In this extraction step, the concentration of soluble solids, minus the amount of insoluble solids in the total solids amount, is preferably extracted until the concentration is between 20 DEG BRIX and 22 DEG BRIX. This is because components such as organic acid are most widely distributed in the above concentration range.

The extract solution is filtered through a filter (about 55 mu m) and then concentrated by a known centrifugal thin film concentrator. In one embodiment, the extraction solution may be concentrated through a centrifugal thin film concentrator rotating at a temperature of 75 to 85 DEG C at 1,000 to 1,400 rpm as a condition for the concentration.

The concentrate formed and concentrated by the concentrator as described above is cooled in a freezer at -30 to -35 캜 for 10 to 14 hours and then lyophilized again at a temperature of about -45 캜 for 46 to 50 hours, And a fine line tree extract in powder form having a particle size of 10 to 200 mesh can be obtained through a known mill.

The term " extract " as used herein generally refers to a crude extract, but in broader terms it also includes fractions fractionated. That is, the bitter gourd extract includes not only those obtained by using an extraction solvent but also those obtained by further applying a purification process thereto. For example, a fraction obtained by passing the above extract through an ultrafiltration membrane having a constant molecular weight cut-off value, a separation by various chromatographs (manufactured for separation according to size, charge, hydrophobicity, or affinity) , Fractions obtained through various purification methods that are additionally included are also included in the scope of the present invention.

The extract may be formulated into various forms depending on the application subject and the purpose of use, such as a pill, a tablet or a capsule. In one embodiment, the extract of Helicobida plant of the present invention is excellent in selective growth inhibition effect and antioxidative activity against microorganisms, and the composition containing the same is useful as a medicament which can treat, prevent or alleviate infectious diseases caused by oxidation and bacteria, Functional foods, cosmetics, food, and feed additives.

In addition, the composition according to the present invention is useful as a medicament for the prevention of food poisoning, a food for preventing food poisoning, a fiber, a sanitary napkin or an antimicrobial pad, a daily necessity requiring antimicrobial activity, May be added and added to the species.

Hereinafter, the present invention will be described more specifically by way of Examples and Experimental Examples.

EXAMPLES Preparation of Extracts Extracted from Leaf Tree Leaves

Leaves and stems were collected from spikelets cultivated in Chougok - ri, Goesan - gun, Chungbuk Province. The collected leaves and stems were washed and dried, and the dried leaves and stems were cut into 5 - 6cm size. The cut leaves and stems were separated from each other and immersed in a 70% (w / w) alcohol solution of 10 times (w / w) and extracted for 24 hours. The extracted solution was filtered with a milipore filter (55 탆), concentrated using a centrifugal thin film concentrator, and used as a sample for the experiment. The obtained samples were stored in a freezer maintained at a temperature of -30 DEG C until the day of the experiment.

[Experimental Example 1] Analysis of Acteoside (verbascoside)

In this experiment, the contents of acteoside (verbascoside) and isoacteoside (isoverbascoside), which are present in the brackish tree, were compared through sequential extracts of Leuconthus japonicus. In addition, direct extraction (reflux), ultrasound, And verbascoside contents by the solvent temperature condition were investigated. The analytical conditions of HPLC for verbascoside content analysis are as follows.

· HPLC condition

Column: Capcell pak C18, 4.6 ml i.d. × 250 mm L., 5 μm, Shiseido, Japan

Flow rate: 1.0 ml / min

Detection: PDA, 330 nm

Mobile phase: (A) 0.1% H ₃ PO ₄ / DW, (B) ACN

· Injection volumn: 10 μl

Gradient: Time (min) A Conc. (%) B Conc. (%) 0 90 10 7 75 25 15 75 25 23 40 60 25 0 100 30 0 100 32 90 10 40 stop

The verbascoside standard curve and spectral analysis results according to the optimal HPLC analysis conditions are shown in FIG. Referring to FIG. 9, when verbascoside was analyzed according to analysis conditions of optimum HPLC, a peak was observed at about 13.8 minutes, and a standard spectrum was observed at 330 nm as a result of wavelength analysis. Based on the results obtained from the above results, the extracts obtained for each condition were analyzed in order to obtain optimal extraction (sequential extraction, direct extraction, ultrasonic extraction, supercritical extraction) and conditions (temperature, solvent%) of the leaves.

The verbascoside content of extracts by sequential extraction method was analyzed by HPLC at 203 nm wavelength. No verbascoside was detected in the hexane layer, and it was detected in ethylacetate layer and methanol layer. In the ethylacetate layer, 2 peaks were overlapped at the same time. As a result of analyzing each peak through spectrum analysis, it was confirmed that peak 1 is verbascoside because the peak 1 is similar to the spectrum of verbascoside standard material.

In the previous experiment, various peaks were analyzed by HPLC at 330 nm wavelength. As a result, verbascoside was not detected in the hexane layer and it was detected in ethylacetate layer and methanol layer.

As a result, two peaks were observed at the wavelength of 330 nm similar to that of the previous 203 nm wavelength analysis and two similar peaks of the ethylacetate layer. As a result of the spectrum analysis, it was found that the first peak was verbascoside.

The verbascoside contents of methanol extracts (50%, 100%) and solvent extracts (50% ethanol extracts) were verbascoside As a result of observing the content, verbascoside content was found to be the highest in 50% ethanol extraction.

In order to observe the verbascoside content of the leaf extracts by direct ultrasonic extraction at 60 ℃, verbascoside contents of the extracted solvents were examined by methanol, ethanol, 50% methanol and 50% ethanol after the extraction. And verbascoside contents were the highest in 50% ethanol extract.

In order to observe the verbascoside content of Leaf Leaf Extract through Direct Reflux Extraction, the extraction solvent was extracted with methanol, ethanol and water, and the verbascoside content of the extract was observed. , Respectively.

In order to observe the verbascoside contents of the supercritical extracts of Leucumber leaf, the extracts obtained by dissolving the extracts obtained by the supercritical process in 1% ethanol and water (DW) were directly analyzed by HPLC analysis.

The extraction method and the verbascoside content of the extraction conditions are summarized in Table 2 below.

Extraction Condition Verbascoside
Mg / 100 ml
extract Solvent Condition
(Temp., Run time) Sequential extraction
Step soxhlet ① Hexane 68 ° C, 8 hr N.D. @ 2 EtOAc 77 ° C, 8 hr 20.86 ③ MeOH 65 ° C, 8 hr 1725.26 Reflux extraction
Reflux ④ MeOH 65 ° C, 2 hr 847.05 ⑤ EtOH 78 ° C, 2 hr 921.60 ⑥ Hot-water 100 ° C, 2 hr 70.77 Ultrasonic extraction
Ultrasound-
assisted ⑦ 50% MeOH Ambient, 30 min 150.43 ⑧ 50% MeOH, 60 캜 60 ° C, 30 min 337.09 ? 50% EtOH Ambient, 30 min 248.37 ⑩ 50% EtOH, 60 属 C 60 ° C, 30 min 422.16 (11) 100% MeOH Ambient, 30 min 236.31 ⑫ 100% MeOH, 60 캜 60 ° C, 30 min 402.25 ⑬ 100% EtOH Ambient, 30 min 332.30 ⑭ 100% EtOH, 60 캜 60 ° C, 30 min 470.02 Supercritical extraction
Sc-CO ₂ - ^* 50 ° C, 280 bar,
180 min N.D. Water soluble layer 0.64

^* Sampling: 10 mg Sc-CO ₂ extract in 1 ml EtOH

The highest verbascoside detection rate was obtained at 60 ℃ ultrasonication (30min) using ethanol (50%, 100%) on the verbascoside by HPLC-PDA.

Based on the above conditions, the contents of verbascoside were analyzed by hot water extraction and alcohol extraction as well as the stem leaf. In order to prepare a sample solution for the analysis of the content of verbascoside leaf, stem verbascoside, 100 ml of solvent was added to 5 g of the analytical sample, and 100 ml of solvent (hexane, ethylacetate and methanol) Stepwise down extraction (8 hours each) was carried out. Afterwards, 100 ml of the solution was filtered through Whatman No. 2 filter. The purified solution was filtered using a sylinge filter (0.45 μm PVDF) The filtrate was used as the test solution.

For direct extraction, 50 ml of solvent was added to 2.5 g of the analytical sample and extracted for 2 hours in 50, 85, 100% methanol and ethanol. Thereafter, the extract solution was filtered using Whatman No. 2, filtered through a 0.45 μm PVDF syringe filter, and the filtrate was used as a test solution.

In the case of ultrasonic extraction, methanol and ethanol (20 ml) were added to 1 g of the analytical sample for 30 minutes, and the extracted solution was filtered using Whatman No. 2 and then diluted to 20 ml. After filtration using a 0.45 μm PVDF syringe filter, the filtrate was used as a test solution.

In the case of leaf and stem hot water extraction, 900 g of water was added to 100 g of powder and extracted at 100 ° C for 2 hours. The extraction yields were 12% and 12% in leaves and 6% in leaves, respectively.

For the extraction of leaves, 12 kg of 70% EtOH was added to 12 kg of powder and extracted at room temperature for 24 hours. The extraction yield was 10.8% (1.3 kg).

[Experimental Example 2] Antimicrobial activity test of Leaf leaf extract

Antimicrobial activity against cavities, vaginitis bacteria, purulent bacteria and food poisoning bacteria

Streptococcus mutans , Candidia albicans , Staphylococcus aureus , E. coli and Salmonella typhimurium used in the strain culture were cultured under the conditions shown in Table 1 below .

turn Microorganism name Use badge Growing conditions Remarks One Escherichia coli
( E. coli ) EMB Agar
(Eosine Methylene Blue Agar) 35 - 37 ° C
Aerobic food poisoning 2 Vaginitis
( Candidia albicans ) PDA
(Potato Dextrose Agar) 25 - 30 ° C
Flowable anaerobic vaginitis 3 Staphylococcus aureus
( Staphylococcus aureus ) TSA
(Tryptic strain Agar) 35 - 37 ° C
Aerobic Purulent dermatitis and food poisoning 4 Cavities
( Streptococcus mutans ) BHI
(Brain Heart Infusion Agar) 35 - 37 ° C
Flowable anaerobic cavity 5 Salmonella
( Salmonella typhimurium ) SS agar
(Salmonella-
Shigella Agar) 35 - 37 ° C
Aerobic food poisoning

In order to analyze the antimicrobial activity of the extracts of Leucocephala leaf extract (hot water, 70% alcohol, butylene glycol) against the Toxins, Vinegars, Staphylococcus aureus, Escherichia coli and Salmonella strains, each microorganism was first cultured under optimal culture conditions according to 1 × ¹⁰ 10 CFU / ㎖ after concentration in liquid culture it was plated on each of the 100㎕ solid medium. Then, a paper disc having a diameter of 8 mm sterilized was placed on a solid medium, and 50 μl of each extract was loaded according to the dilution drainage. The antimicrobial activity was measured by measuring the diameter (unit: mm) of the clear zone (growth inhibition circle) after culturing the solid medium in an incubator for 48 to 72 hours at the optimum temperature. The results of the respective tests are shown in Tables 4 to 11 and Figs. 1 to 4 below.

Antimicrobial Activity of Solanum sp. Item division Diameter (mm) A Control
(DMSO) 8.5 B green tea
(Butyl 1/10) 10.7 C Leaf tree leaves
(1/10 of heat) 13.9 D Leaf tree leaves
(Butyl 1/10) 15.1 E Leaf tree leaves
(EtOH 1/10) 16.3

Antimicrobial activity of extracts from the leaves Item division Diameter (mm) A Control
(DMSO) 8.5 B green tea
(Butyl 1/10) 8.4 C Leaf tree leaves
(EtOH 1/10) 12.1 D Leaf tree leaves
(EtOH 1/20) 8.5 E Leaf tree leaves
(EtOH 1/40) 8.5

Antimicrobial Activity of Solanum sp. Item division Diameter (mm) A Control
(DMSO) 8.5 B green tea
(Butyl 1/10) 9.5 C Leaf tree leaves
(1/10 of heat) 8.5 D Leaf tree leaves
(EtOH 1/10) 9.0 E Leaf tree leaves
(Butyl 1/10) 9.2

Antibacterial Activity of Butyleneglycol Leaf Extract of Leaf Leaf on Vinegar Bacteria Item division Diameter (mm) A Control
(DMSO) 8.5 B green tea
(Butyl 1/10) 10.1 C Leaf tree leaves
(Butyl 1/10) 13.5 D Leaf tree leaves
(Butyl 1/20) 8.5 E Leaf tree leaves
(Butyl 1/40) 8.3

Antimicrobial Activity of Leaf Extract Solvent on Staphylococcus aureus Item division Diameter (mm) A Control
(DMSO) 8.5 B green tea
(Butyl 1/10) 9.5 C Leaf tree leaves
(1/10 of heat) 11.1 D Leaf tree leaves
(Butyl 1/10) 9.1 E Leaf tree leaves
(EtOH 1/10) 15.9

Antimicrobial Activity of Spirulina Leaf Extracts on Staphylococcus aureus Item division Diameter (mm) A Control
(DMSO) 8.5 B green tea
(Butyl 1/10) 8.5 C Leaf tree leaves
(EtOH 1/10) 12.9 D Leaf tree leaves
(EtOH 1/20) 10.0 E Leaf tree leaves
(EtOH 1/40) 8.7

Antibacterial activity of Escherichia coli on Leaf Extract Solvent Item division Diameter (mm) A Control
(DMSO) 8.5 B Leaf tree leaves
(EtOH 1/10) 12.8 C Leaf tree leaves
(1/10 of heat) 9.9 D green tea
(Butyl 1/10) 9.1

Antimicrobial activity of Salmonella sp. Item division Diameter (mm) A Control
(DMSO) 8.5 B Leaf tree leaves
(1/10 of heat) 8.5 C Leaf tree leaves
(EtOH 1/10) 9.6 D green tea
(Butyl 1/10) 9.4

As shown in Table 4 to Table 11 and FIG. 1 to FIG. 4, it was confirmed that the extract of Leucumber leaf had antimicrobial activity against Toxins, Nitrification bacteria, Staphylococcus aureus, Escherichia coli and Salmonella, The antimicrobial activity of Leuconthus sp.

In the case of the extract of 70% of the leaves of Leucumber leaf, the antimicrobial activity of the microorganism was found to be excellent in terms of efficacy. However, when the concentration of the extract was diluted 1/10 times or more, Significant results have also been obtained that at least 10% should be added to the product in order to have a definite effect as an antimicrobial agent.

[Experimental Example 3] Antioxidant activity test using Leaf leaf extract

1. DPPH radical scavenging ability

Radical scavenging activity of 1,1-diphenyl-picryllhydrazyl (DPPH, Sigma-Aldrich Co.) was determined by adding 100 μl of a 0.1 mM DPPH solution dissolved in methanol to the extracts of Pinus rigida (6.25, 12.5, 25, 50 and 100 ㎍ / ㎖) was added to each well and incubated in the dark room for 30 minutes. Then, the absorbance was measured at 517 nm using a microplate reader.

Here, A sample represents the absorbance of the sample and the DPPH reaction solution, A blank1 represents the single absorbance of the sample, A blind represents the single absorbance of the DPPH solution, and A blank2 represents the blank sample.

On the other hand, to investigate the radical scavenging ability of the extracts of spruce leaves, the scavenging ability of DPPH radicals, which are relatively stable radicals, was investigated. As a result of observing the color reaction of purple DPPH radicals at the stable stage by mixing the DPPH radicals with the extracts of the spikelet leaves (6.25, 12.5, 25, 50 and 100 ㎍ / ㎖) The DPPH radicals were decreased, and the results were shown in FIG. 5 as a low-performance percentage graph.

2. ABTS radical scavenging ability

To measure the radical scavenging activity, 7.4 mM ABTS and 2.6 mM potassium persulfate (Sigma-Aldrich Co.) were dissolved in 24 (v / v) acetonitrile (Sigma-Aldrich Co.) After incubation for an hour to form ABTS radicals, the solution was diluted with distilled water to an absorbance value of 1.5 at 760 nm. After diluting ABTS solution (20 μl) with the extracts of the vinegar extracts (62.5, 125, 250, 500 and 1,000 μg / ml) prepared by concentration, the absorbance was measured at 760 nm using a microplate reader.

ABTS radicals are known to be highly stable in aqueous solution similar to DPPH radicals. In order to investigate the scavenging ability of DPPH radical scavenging ability and other types of radicals in the extracts of Sprague-Dawley leaves, The radical scavenging ability was investigated. As a result of observing the color reaction of ABTS radicals showing blue color at stable stage by mixing the DPPH radicals with the extracts of spikelet leaves (62.5, 125, 250, 500 and 1,000 μg / ml) The ABTS radicals were decreased in a concentration-dependent manner, and the results are shown in FIG. 6 as a low-performance percentage graph.

3. Measurement of FRAP Capability

100 μl of 0.2 M sodium phosphate buffer (pH 6.6) and 100 μl of 1% potassium ferricyanide (Sigma) were added to 100 μl of Leucon leaf extract at different concentrations (0.25, 0.5, 1 and 2 mg / 100 μl of trichloroacetic acid (Sigma-Aldrich Co.) was added to each well, and the mixture was centrifuged at 12,000 rpm for 10 minutes. After that, 20 μl of 0.1% ferric chloride (Sigma-Aldrich Co.) was added, reacted for 20 minutes at 37 ° C in a constant temperature water bath, and the reducing power was measured at 700 nm using a microplate reader.

To investigate the antioxidant activity of Leuconostoc sp. Leaf extract, the reducing power on the inhibition of iron oxidation in the iron oxidation (Fe ^{2 +} → Fe ^{3 +} ) model, . As the concentration of the treatments was increased, the reducing power was increased when treated with the extracts of 62.5, 125, 250, 500 and 1,000 μg / The effect was observed to be high in the concentration. Fig. 7 shows the results of the FRAP performance of Leucocephala leaf extract.

4. Reducing power measurement

Reducing power was determined by adding 1 ml of 200 mM sodium phosphate buffer (pH 6.6) and 1 ml of 1% potassium ferricyanide (Sigma) to 1 ml of lyophilized broth extracts of 31.25, 62.5, 125, 250 and 500 μg / -Aldrich, USA). The mixture was incubated at 50 ° C for 20 minutes, then 1 ml of 10% trichloroacetic acid (w / v) was added, centrifuged at 3,000 rpm for 10 minutes, and 1 ml of distilled water and 0.1% ferric chloride ), And the absorbance was measured at 700 nm. The results of the reducing power performance test of Leuconostoc spider leaf extract are shown in FIG.

As a result of the study on the reducing power ability of the extracts of Leuconostoc sp. Leaves, the extracts of Leuconostoc sp. Leaves were found to increase Reducing power ability in a concentration dependent manner.

Claims (10)

An antimicrobial composition comprising, as an active ingredient, an extract extracted from any one or two or more sites selected from leaf, stem, fruit, roots, husks and seeds of a spiny tree. The method according to claim 1,
The above-
The present invention relates to a method for preparing a pulverized product by pulverizing at least one selected from leaf, stem, fruit, roots, husks and seeds of a bracken tree to prepare a pulverized product, mixing water or any one of ethyl alcohol and ethanol And allowed to stand at room temperature for 1 to 2 days,
Wherein the extract is prepared under heating, reduced pressure and vacuum. The method according to claim 1,
The above-
But are not limited to, water, purified water, C ₁ -C ₄ alcohols, glycerin, acetone, hexane, ethyl acetate, butylene glycol, propylene glycol, dichloromethane ), Chloroform (chloroform), and ethyl ether, or a mixture of two or more thereof. 4. The method according to any one of claims 1 to 3,
Wherein said composition is in the form of a pharmaceutical composition for treatment. 4. The method according to any one of claims 1 to 3,
Wherein said composition is in the form of a health functional food. 4. The method according to any one of claims 1 to 3,
Wherein said composition is in the form of a cosmetic composition. 4. The method according to any one of claims 1 to 3,
Wherein said composition is in the form of a food additive. 4. The method according to any one of claims 1 to 3,
Wherein said composition is in the form of a feed additive. 4. The method according to any one of claims 1 to 3,
Wherein the composition is added to at least one member selected from the group consisting of medicines for the prevention of food poisoning, foods for the prevention of food poisoning, fibers, daily necessities, building materials and household articles. The method according to claim 1,
Wherein the active ingredient comprises one or both components selected from the group consisting of verbascoside and polyphenol.

Images