KR100775395B1 - Compounds for red tide prevention comprising natural plant extracts - Google Patents

Abstract

Compositions for prevention of red tide comprising natural plant extracts are provided to control and prevent red tide selectively by killing red tide-causing bacteria in an environment-friendly manner, thereby replacing chemical agents and loess for preventing red tide. A composition for prevention of red tide by inhibiting growth of Cochlodinium polykrikoides comprises the extract of at least one natural plant which is selected from Sophora flavescens Ait, sweet flag and Eugenia caryophyllata and is prepared by extracting natural plants with water, hydrophilic organic solvent or a mixed solvent thereof, and further comprises pharmaceutically acceptable diluents or excipients.

Description

Red tide prevention comprising natural plant extracts as an active ingredient {compounds for red tide prevention comprising natural plant extracts}

1 is a photograph of death of coclodinium red tide using the iris extract according to the present invention.

Figure 2 is a photograph of death of coclodinium red tide using the clove extract according to the present invention.

Figure 3 is a photograph of death of coclodinium red tide using the ginseng extract according to the present invention.

Figure 4 is a photograph of the death of coclodinium red tide by the loess spray according to the present invention.

The present invention relates to a red tide control composition comprising a natural plant extract as an active ingredient, and more particularly to a red tide control composition containing any one or more extracts of red ginseng, irises and cloves as an active ingredient.

The present invention has been developed worldwide, the development of the control agent of the red tide source that causes enormous economic damage every year in the aquaculture industry site, Wawa imitation algae which is the causative organism of red tide using natural extracts of ginseng, iris and clove, which are derived from natural products ( kokeulrodinium belonging to渦鞭毛藻類) (Cochlodinium Polykrikoides ) to the algae effect, to develop a product that is excellent in the environment-friendly and biodegradable red algae control agent in the present invention.

Red tide phenomena occur on coasts around the world, and the number of occurrence areas and the number of occurrences not only in Korea but also in many countries continues to increase, and the economic damage of the aquaculture industry due to red tide is increasing year by year. With the development of industry, along with coastal inflow of various pollutants, domestic and industrial wastewater, which is rich in perishable organic pollutants and growth promoting substances, such as wastewater containing nitrogen (N) and phosphorus (P), flows into the sea. The frequency of red tide caused by flagellar algae is increasing rather than red tide due to sea eutrophication and increased silicate. In addition, as the frequency of red tide caused by flagella algae increases, the red tide phenomenon is increasing and prolonged, and the frequency of red tide caused by harmful flagella algae is increasing.

In general, species that occur mainly in the coast of Korea and produce toxic substances that directly kill fish or shellfish are species belonging to the genus of braids such as Cochlodinium , Gymnodinium , Gyrodinium , Heterosigma . Among them, the Cochlidinium genus is a phytoplankton that lurks in the state of dormant spores in seawater, but starts its activities when the water temperature rises above 23 ° C and becomes rich in nutrients. Photosynthetic action, but because of the movement of the vortex with flagella, is also characterized by animal characteristics. One or several cells are connected and the size of the cells is 20 to 40 μm (1 μm is 1 millionth of a meter), although they are not toxic on their own. Can be lethal by breathing difficulties. In the coast of Korea, fish and shellfish have been killed since 1989. Especially, from 1990 to 1993, fish and shellfish have been killed in large quantities from August to October and from 1995 to October in 1995 from Chungmu coast, Yonsei ~ Hakrimdo, Sayangdo, and the South Seas Lethal. Currently, the main areas of origin are Goheung Na-ho, Yeosu Sea Bay, Namhae, Tongyeong, Busan Coast, and South East Sea.

In order to cope with the enormous economic damages of the aquaculture industry caused by these harmful red tide organisms, many researches are being actively conducted to control the red tide phenomenon. There are physical, biological and chemical control methods for removing and removing red tide organisms for controlling harmful red tide organisms. Representative examples include air injector or filtration device, ultrasonic treatment and ozone 153 treatment method, filtration or centrifugation of red algae to recover from the sea surface, clay or polymer flocculant to adsorb, aggregate and precipitate red tide organisms. Numerous methods such as methods using natural enemies or bacteria of red tide-inducing organisms, chemicals that destroy the cells of red tide organisms (e.g., hydrogen peroxide, copper sulfate, chlorine, herbicides, etc.) and natural or derivatives thereof. There is. Among the control methods, the relatively high effect is the clay adsorption method using ocher, which utilizes the property of spraying clay in seawater to adsorb and settle the red tide organisms, and to destroy the cells of the red tide organisms by eluting aluminum ions in the clay. This method was mainly studied in Japan and has been conducted in Korea since 1985 by the National Fisheries Research and Development Agency. However, the ocher spray method is non-selective, and when the flow of algae is strong, the control effect is lowered, and red tide occurs again when the amount of sunshine and water temperature increase. Existing methods of red tide control also suffer from disadvantages such as non-selective red tide, tidal and water temperature, safety against non-target organisms, environmental pollution, human toxicity, and difficulty in mass production. Therefore, the development of environment-friendly technology to compensate for these shortcomings is required.

Therefore, the present inventors have made a thorough research to overcome the problems of the prior art, in the case of red tide control composition containing any one or more extracts of red ginseng, iris and clove as an active ingredient, as a pesticide of harmful tide organisms It was confirmed that it can be applied as an eco-friendly control agent to replace various red tide control agents used when red tide occurs, thus completing the present invention.

The present invention, in order to solve the limitations and side effects of using the red tide control agent used in the red tide organisms shown above, by searching for a material using a proven natural material, in particular, a material having an excellent killing effect to the harmful algae Based on the results of the search and research, it is possible to prove the killing effect of harmful red algae on the extract of red ginseng, iris and clove, and to improve the industrial added value and to completely exclude the dysfunction by the use of chemicals. The purpose is to develop the product. In order to achieve the above object, the present invention takes extracts of red ginseng, irises and cloves that have anti-bacterial and antibacterial activity, to identify the biological activity of red tide-causing organisms, and to identify the killing effects of red tide organisms and to produce products suitable for economic efficiency. There is a purpose.

According to an aspect of the present invention, the present invention provides a red tide control composition containing an extract of any one or more of red ginseng, irises and cloves as an active ingredient.

The ginseng (苦 蔘, Sophora flavescens Ait) used in the present invention belongs to the legumes (Leguminosae) and is a perennial plant is Korea, China, Japan and the like. Commonly used ginseng root, the main components include several kinds of quinolizidine, alkaloid, flavonoid, saponin. In oriental medicine, it has been used for diuretic, antipyretic, antibacterial and anti-Trichomonas action, clinically used for eczema, dermatitis, skin disease, and is known to be effective in bacterial and enteritis. Iris (菖蒲, sweet flag) belongs to Chunnamseong Province (Araceae), and several seaweed plants have a fragrant substance throughout the roots, leaves and roots. Common irises use roots, which contain essential oils, tannins, starch, palmitic acid and vitamin C. Essential oils include asarone, eugenol, calameone and sesquiterpene. Iris containing these ingredients is used as a folk remedy, for example, to help digestion by bringing out gastric juice and hydrochloric acid in the stomach. Diarrhea, inflammation of the ears, skin disease, athlete's foot, analgesic, anticonvulsions, antitussive expectoration, hypothermia, Efficacy in blood circulation, and used in the West as an insect repellent, tonic, and fragrance raw material (Information Seop, Shin Min Gyo, Ph.D., Yeonglimsa, pp278-280, 1998). Also known as Evergreen, Clove (丁香, Eugenia caryophllus) is a pink flower that is the source of clove. Cloves include Eugenol, acetyl eugenol, α-caryophyllene, beta-cario, ethylene, methyl-n-amiketone, methyl-n-heptylketone, sesquiterpene, furfural, wanlin, tannin, oil, lead and oleanolic acid. It is contained. Clove has anti-inflammatory, analgesic, antithrombotic, and digestive system effects, and is used as a fragrance, antibacterial, cleaning agent, insect repellent, anesthetic, antiseptic, soap, and rust inhibitor.

In the present invention, the extract of red ginseng, irises and cloves may be extracted with water, a hydrophilic organic solvent or a mixed solvent thereof, but is preferably extracted with a hydrophilic organic solvent. Here, the hydrophilic organic solvent may be used diluted in water to 50 to 100%.

In the present invention, examples of the hydrophilic organic solvent include lower alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol and isopropyl alcohol; Lower aliphatic ketones having 2 to 5 carbon atoms such as acetone and methyl ethyl ketone; Lower polyhydric alcohols having 2 to 5 carbon atoms such as 1, 3-butylene glycol, propylene glycol, and glycerin, and the like, and these have a hydrophilic alcohol group (OH) or a ketone group (COC) and thus have a similar polarity. Similar extractive activity for

In the present invention, the extraction method of the extract may be used a natural product extraction method of a commonly used plant, preferably may be used in addition to organic solvent extraction method, hot water extraction and steam distillation method. Hot water extraction is performed for 30 minutes to 2 hours at 60 ~ 100 ℃ depending on the state of the sample. Extracts extracted with hot water are usually centrifuged, take only supernatants, and lyophilize to prepare powders, verify biological activity or use them as commercialized products. In addition, the steam distillation method puts ginseng, irises and cloves itself or dried and ground materials into an extractor, and distills water, and when steam is mixed into the extractor, the active ingredient of the sample is extracted by the mixed steam. Only the oil layer is separated from the water layer to obtain essential oils. The extraction time is appropriate for 1 hour 30 minutes to 2 hours depending on the sample state.

In the present invention, the red tide control composition may further comprise a pharmaceutically acceptable diluent or excipient, in addition to the extract of red ginseng, green vesicles and cloves having an algic effect as an active ingredient.

In the present invention, the red tide control composition is preferably characterized by having a control effect on Cochlodinium polykrikoides . In the embodiment of the present invention, the algae effect on the red tide-inducing organism was tested.

When described in detail by the accompanying table as follows.

The main components of natural ginseng, irises and cloves of the present invention are shown in Table 1.

Table 1. Natural substances Gosam , Iris, cloves  Main ingredient of the extract

Kinds main ingredient Gosam saponin, sapogenol, sophoraflavoside, lupine alkaloids, (+)-isomatrine, kushequinone A, kurarinol, kushenolA, B, E, F, kushenol J, K, L, M calamus tannin, vitamin C, acorin, palmitic acid, starch, eugenol, asarylaldehyde, asarone, shyobunone, epishyobunone, isoshyobunone, calamendiol, isocalamendiol, calacone, acorone, acoronene, acorenone, α-pinene, camphene, camphene , borneol, calamene, caryophyllene, elemene, curcumin, selinene, acolamone, isoacolamone. cloves Eugenol, eugenyl acetate, caryophyllene, isocaryophyllene, isocaryophyllene, acetyl eugenol, α-caryophyllene, beta-cario, lene, methyl-n-amiketone, methyl-n-heptylketone, sesquiterpenes, furfural, warnilin, tannin, oil, Lead, oleanolic acid

The main action of these components is shown in Table 2. As shown in Table 2, the action of the main components of the natural substance extract is composed of substances that exhibit antifungal, antibacterial and antiparasitic activity, and is also configured to have an inhibitory and killing effect on harmful algae.

Table 2. Natural substances Gosam , Iris, cloves  Main action of extract

main ingredient Main action matrine Anti-ulcer, antipyretic, gastrointestinal contraction flavonoids Antibacterial, antifungal asarone (C ₁₂ H ₁₆ O ₃ ) Strong antiviral and antibacterial effect, antiseptic effect, blood circulation promotion, blood pressure increase effect, tonic effect eugenol (C ₁₀ H ₁₂ O ₂ ) Antioxidant, antibacterial, antiseptic, antifungal, otitis, antiviral, antiseptic

In addition, one of these extracts contains major ingredients such as shyobunone, calamene, acorone and camphene. In general, the antifungal and antimicrobial effects of essential oils consist mainly of phenylpropane and terpine. It is determined by the activity and content of oxides, esters, alcohols, phenols, aldehydes, ketones, monoterpines, sesquiterterpines and the like. The components of these essential oils are not just molecular, but have a very complex structure and have been shown to have antiviral, antibacterial, antifungal, and antiworm activity.

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

Example  1. Harmful organisms Algae effect  Preparation of Extract

Extracts of natural ginseng, irises and cloves used in the present invention are sun-dried flowers, rhizomes and rhizomes from a solvent group consisting of water, ethanol, isopropyl alcohol, methanol, propanol, butanol, glycerin, butylene glycol An extract obtained using one or more solvents selected. The harmful algae killing effect extract of the present invention is contained in the range of 1 to 50% by weight of the extract obtained above.

Example  2. Gosam , Iris, cloves  Experimental method of red tide control effect of extract

In order to confirm the control effect of red ginseng, irises and cloves extract on red tide-induced red tide organisms, the red ginseng, irises and clove extracts extracted by the extraction method of Example 1 are causing the biggest problems in the current red tide-inducing organisms in Korea. known kokeulrodinium poly Cri Koh des (Cochlodinium polykrikoides ) was confirmed for the control effect.

First of all, natural ginseng, irises, fennel extracts and ocher were compared and tested. The control effect of red tide-causing organisms was tested using the 12 well plate method.

Cochlodinium ( Cochlodinium) polykrikoides ) were cultured in F / 2 medium (Guillard & Ryther 1962, Guillard 1975) prepared by seawater from the National Fisheries Research and Development Institute under 25 ° C., photoperiod 24 hours, pH 7.5, and light intensity 2500 lux.

Cultured Cochlodinium 1 g each of polykrikoides was added to 12 well plates, and the extract of red ginseng, irises and cloves prepared in Example 1 was administered. Each concentration was administered in the range of 0.001% to 5.0% compared to the culture medium added coclodinium red tide. Coclodinium red tide without the extract was used as a control.

Ocher has been used as a substance to rescue red tide by focusing on the colloidal particles' ability to coagulate and adsorb suspended solids (nutrients and microplankton) in seawater. In other words, the principle of remedy is to agglomerate and settle the red tide organisms in the red soil when red tide occurs. The ocher, which is known for its excellent removal effect of coclodinium red tide, was also compared with natural extracts. After the ocher was sufficiently diluted in F / 2 medium prepared with seawater, it was administered in the range of 0.005% to 2.5% compared to the culture medium added coclodinium red tide. Similarly, coclodinium red tide without the extract was used as a control.

Example  3. Gosam Experimental Method of Red Tide Control Effect in the Mixing Treatment of Water and Iris Extract

Example 1. The extract of the iris and Sophora flavescens extract the one manufactured by: administering to the addition of the culture medium compared to 0.001% ~ 2.0% of the red tide organisms kokeulrodinium culture is mixed, in a ratio Cochlodinium The interaction with polykrikoides was confirmed. Coclodinium red tide without the mixed extract was used as a control.

Example  4. Gosam , Irises and cloves  Experimental Method of Red Tide Control Effect in Mixed Treatment of Extracts

Example 1 The extract of Sophora flavescens, iris, and the clove extract prepared from 1: 1: 1 were mixed at a ratio by administering to the culture solution was added over 0.001% ~ 2.0% of the red tide organisms cultured kokeulrodinium Cochlodinium The interaction with polykrikoides was confirmed. Coclodinium red tide without the mixed extract was used as a control.

Experimental Results 1. Red tide-causing organisms by concentration of natural substance extract and yellow soil Algae effect  Verification

Cochlodinium Cochlodinium polykrikoides ) was added to a 12well plate and the results of the experiments with the administration of each natural substance and ocher are shown in the following Tables 3, 4, 5 and FIGS. 1, 2, and 3.

Table 3. Effect of Controlling Coclodinium Red Tide Using Iris Extract

density(%) calamus 0.001 0.005 0.01 0.02 0.03 0.04 0.05 0.1 0.2 0.5 1.0 2.0 effect - - - - - - ++ +++ +++ +++ +++ +++

(+++: kills immediately, ++: kills within 10-20min, +: kills within 1hr,-: does not kill)

Table 4. cloves  Control of Coclodinium Red Tide Using Extracts

density(%) cloves 0.1 0.15 0.2 0.25 0.5 1.0 2.0 5.0 effect + ++ ++ ++ ++ +++ +++ +++

(+++: kills immediately, ++: kills within 10-20min, +: kills within 1hr,-: does not kill)

Table 5. Gosam  Control of Coclodinium Red Tide Using Extracts

density(%) Gosam 0.001 0.01 0.05 0.08 0.1 0.2 0.3 0.4 0.5 effect - - - + +++ +++ +++ +++ +++ Gosam 0.6 0.7 0.8 0.9 1.0 2.0 3.0 4.0 5.0 effect +++ +++ +++ +++ +++ +++ +++ +++ +++

(+++: kills immediately, ++: kills within 10-20min, +: kills within 1hr,-: does not kill)

Table 6. Effect of Coclodinium Red Tide Control on Spraying Yellow Clay

density(%) ocher 0.005 0.01 0.02 0.04 0.08 0.16 0.32 0.64 1.25 2.5 effect - - - - - + ++ +++ +++ +++

In the case of ocher, it is not based on the degree of coarse but on the degree of coagulation.

Table 3, 4, 5 and coclodinium red algae treated with red ginseng, irises and clove extracts respectively were immobilized and killed in the case of red ginseng extracts, and the iris and clove extracts were lysed and destroyed. It can be seen that. In addition, these natural extracts can be seen that the killing effect is very excellent when compared with the yellow soil of Table 6 and FIG.

Experiment Result 2. Gosam Red tide-causing organisms when mixed with natural iris extracts Algae effect  Verification

Experimental results showed that red ginseng and irises were effective in controlling red tide at lower concentrations, as shown in Experiment 1. Accordingly, the results of the experiments in which red ginseng and irises were mixed in a 1: 1 ratio by adding Cochlodinium polykrikoides to a 12 well plate were shown in Table 7 and FIG. 5.

Table 7. Iris Gosam  Controlling Coclodinium Red Tide by Addition of 1: 1 Extract

density(%) Irises + Gosam 0.001 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09 0.1 effect - - - - - - + ++ ++ ++ +++ Irises + Gosam 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 2.0 effect +++ +++ +++ +++ +++ +++ +++ +++ +++ +++

(+++: kills immediately, ++: kills within 10-20min, +: kills within 1hr,-: does not kill)

Experiment Result 3. Gosam , Irises and cloves  Red tide-causing organisms when mixed with natural substance extracts Algae effect  Verification

As shown in Experiment 1, all extracts of red ginseng, irises and cloves were effective in controlling red tide at lower concentrations. Thus, Cochlodinium polykrikoides was added to 12 well plates, and red ginseng, irises and cloves were mixed in a 1: 1: 1 ratio and then administered to red tide, as shown in Table 8 below.

Table 8. Iris, Gosam  And cloves  Controlling Coclodinium Red Tide by Additive Extract 1: 1: 1

density(%) Irises + Ginseng + Cloves 0.001 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09 0.1 effect - - - - - - - - + ++ +++ Irises + Ginseng + Cloves 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 2.0 effect +++ +++ +++ +++ +++ +++ +++ +++ +++ +++

(+++: kills immediately, ++: kills within 10-20min, +: kills within 1hr,-: does not kill)

In the results shown in Table 7 and Figure 5 can be seen that the coclodinium red tide treated by mixing the ginseng and iris extract 1: 1 mixed cells even when administered at low concentrations. In addition, it can be seen that these natural extracts have excellent killing effects even at very low concentrations when compared with loess. In addition, in Table 8, even when all the three extracts were mixed in the same ratio, the algae effect appeared at a much lower concentration than when compared with ocher.

As such, extracts of red ginseng, irises and cloves according to the present invention are representative of red tide-inducing organisms, coclodinium polycricoides ( Cochodinium). Polykrikoides ) is very effective for killing or inhibiting proliferation within a short time, and it is considered to be a very useful red tide control agent to supplement the disadvantages of the red tide control agents that may occur additionally and to be environmentally friendly and stable.

As described above, according to the present invention it can be seen that the extract of red ginseng, irises, and cloves as an effective ingredient of the red tide control composition. This invention is expected to replace the chemicals and loess used as a red tide control agent in the aquaculture industry, extracts using natural materials, and is expected to have high economic added value. In addition, by developing an environment-friendly red tide control agent containing such an extract can be used as a useful aspect to prevent and remove the enormous damage of red tide that occurs annually worldwide.

Claims (4)

Red tide control composition containing an extract of any one or more of red ginseng, irises and cloves as an active ingredient. The red tide control composition according to claim 1, wherein the extract is extracted with a hydrophilic organic solvent. The red tide control composition of claim 1, wherein the composition further comprises a pharmaceutically acceptable diluent or excipient. According to claim 1, Cochlodinium ( Cochlodinium) polykrikoides ) Red tide control composition characterized in that it has a control effect.

Images