KR101024606B1 - Natural product composition comprising anthraquinone- based compounds for enhansing control of harmful algae - Google Patents

Abstract

The present invention provides a composition for controlling harmful algae containing at least two of anthraquinone-based compounds, preferably at least two of chrysopanol, emodin, fish, lane and aloe-emodine, and harmful algae using the same The present invention relates to a control method, and the mixed composition of the present invention exhibits selective growth inhibitory effect on harmful algae such as southern algae and red algae, and thus the composition for controlling the composition is useful as an environmentally friendly and highly effective harmful algae control agent. Can be utilized.

Harmful algae, cyanobacteria, chrysopanol, emodine, fishion, plant extract fractions

Description

Mixed composition for control of harmful algae containing anthraquinone compound derived from natural products and control method using the same {NATURAL PRODUCT COMPOSITION COMPRISING ANTHRAQUINONE-BASED COMPOUNDS FOR ENHANSING CONTROL OF HARMFUL ALGAE}

The present invention provides a composition for controlling harmful algae containing at least two kinds of anthraquinone compounds derived from natural products, preferably at least two of chrysopanol, emodin, fish, lane and aloe-emodine and harmful algae using the same It is about a control method.

Harmful algal blooms (HABs) are causing various industrial problems, depending on their habitat. The outbreak of harmful algae in lakes, streams, reservoirs, fish farms, etc., 1) causes aquatic deaths (Duke et al . , Weed Sci. 50: 138-151, 2002); 2) It produces off-flavor substances and degrades the quality of drinking water and farmed fish. For example, in the habitat of the harmful algae Oscillatoria perornata flavors) (Duke et al . , Weed Sci. 50: 138-151, 2002); 3) Toxins that are harmful to humans and animals are produced, for example, some birds, such as the genus Microcystis , Nodularia , Anabaena , and Aphanizomenon . Are known to produce toxins that are harmful to humans and animals, respectively, microcystins, nodularin, anatoxin and saxitoxin (Haider et al., Chemosphere 52: 1-21, 2003); 4) causing discomfort and impeding leisure and industrial activity due to the coloring of water and the formation of scum; 5) Over-treatment of chlorine is necessary due to filter paper closure and inhibition of flocculation sedimentation during water treatment, causing economic loss.

On the other hand, in freshwater pavement, the occurrence of algae induces bulge formation, which leads to a significant decrease in the rate of hair growth, and lowers soil temperature, thereby reducing crop productivity. On golf course fairways, grass is often killed due to the occurrence of soil algae. In addition, in the construction and industrial facilities, harmful algae can cause various damages such as water pollution, machine malfunctions, aging, and aesthetic damage.

Therefore, in order to effectively cope with the problem of generation of harmful algae, physical, biological or chemical methods have been tried from various angles, but there are no satisfactory results. In particular, in the case of chemical control, which is considered to be the most practical method, there have been many researches on algae removal after generation after the flocculant. On the other hand, among the compounds used, there are a lot of compounds which are preferable to be replaced in the future because the cost and control effects are relatively emphasized rather than consideration of phosphorus toxicity or environmental stability. For example, copper sulfate, chlorine, alkaline earth metal peroxides are fast-acting and have a pronounced treatment effect but are non-selective, and for copper (Cu) heavy metal accumulation (Hullebusch et al. Water, Air, and Soil Pollution 150: 3-22, In the case of chlorine, it increases the possibility of generating carcinogenic trihalomethanes by interacting with organic substances (humic acid, fulvic acid, etc.) in water. In addition, organic synthetic herbicides, such as simazine and diuron ( Aquaculture 163: 85-99, 2004; Aquaculture 233: 197-203, 2004), may in some cases exhibit toxicity to non-target organisms. Safety is an issue.

Therefore, in the future, it is necessary to minimize the damage to harmful algae while protecting the environment. To this end, there is a need for a technology that suppresses the initial occurrence of algae in a preventive manner rather than after the outbreak of algae using safer compounds. In other words, by searching for environmentally friendly and safer biologically-derived "harmful algae control agents (natural / biochemicals)" (Schrader KK, ACS Symposium Series 848: 195-208, 2003), it is possible to quickly prevent the generation of harmful algae. It is necessary to use as a control agent. Biochemical agents or natural products reported in the literature of properties that may serve this purpose include lysine and its derivatives (Hehmann et al., J. Appl. Phycol . 14: 85-89, 2002), ferulic acid. , Trans -cinnamic acid, quinone compound (Schrader et al . , Bull Environ.Contam.Toxicol . 60: 651-658, 1998), basilyl (bacillamide, Jeong et al . , Tetrahedron Lett. 44: 8005-8007, 2003) , Fischerellin A and B, Hagmann et al . , Tetrahedron Lett. 37: 6539-6542, 1996; Papke et al . , Tetrahedron Lett. 38: 379-382, 1997), oxidized fatty acids and Potassium ricinoleate (Duke et al . , Weed Sci. 50: 138-151, 2002), L-2- azetidinecarboxylic acid (Kim et al., Aquatic Botany 85: 1-6, 2006), rutacris Rutacridone epoxide (Meepagala et al., Phytochemistry 66: 2689-2695, 2005), Ethyl 2-methylacetoacetate (Li & Hu, Appl. Environ.Microbiol. 71: 6545 -6553, 2005), others (Duke et al . , Weed Sci. 50: 138-151, 2002; Schrader et al., Aquaculture 163: 85-99, 1998), and plants include ripening barley straw and Lolium perenne (Duke et al., Weed Sci . 50: 138-151, 2002; Schrader, Agric. Res. 51:21, 2003). On the other hand, as a natural product for the algae patented or published in Korea, the oleander plant extract (Korean Patent No. 456467), the perennial plant or its extract (Korean Patent No. 621917), azetidine derivative (Korean Patent No. 592798), Organic acid (Korean Patent No. 231912), Prodigiosin (Korean Patent No. 661175), Cypress (Korean Patent Publication No. 2006-0103310), Naturally Aging Enzyme Solution of Various Natural Materials (Korean Patent Publication No. 2005-0122919 No.), Bacillus culture filtrate (Korean Patent No. 610527), natural minerals and various plants (Korean Patent No. 623993), Myeongja tree fruit (Korean Patent No. 543747).

However, they tend to have lower overall control efficiency than using copper lactate, hydrogen peroxide, or organic synthetic compounds, which may differ from compound to compound but need to be compensated for practical use, ie, low activity, narrow spectrum of action, and low price. Expensive and other problems have appeared (Duke et al ., Weed Sci. 50: 138-151, 2002). Therefore, technologies that can compensate for these shortcomings must be continuously developed, and as one of the methods, a technique for developing and applying a mixture having synergy with each other is required.

On the other hand, studies to use the fraction of the plant extract or the pure separation component itself as an algae component is varied as described above, but it is confirmed that there is a synergistic effect when mixing the different fractions and in a suitable ratio There is no case of remixing. In addition, it has been reported that anthraquinone compound single components ( Crysophanol , Fishion , etc.) have algicidal activity (Cantrell et al., ARKIVOC 7: 65-75, 2007; US Patent Publication 2003 / 0012804A1; Schrader et al., Bull Environ.Contam.Toxicol . 66: 801-807, 2001), but recently reported that synoxol and fish were synergistic in controlling powdery mildew in cultivated crops (Yang et al . Pest Manag. Sci. 63: 511-515, 2007), synergistic effects on the control of noxious algae between two compounds of the same family (eg, chrysophanol and emodine) have not been reported.

An object of the present invention is to find a mixture of substances having a synergistic effect with each other, in order to control harmful algae more effectively with a low phosphorus toxicity and a nature-friendly substance instead of synthetic organic compounds or inorganic substances which are a problem in safety. To provide a method for manufacturing and controlling harmful algae using the same.

More specifically, it is an object of the present invention to provide a new composition for controlling harmful algae, and a method for controlling harmful algae using the same, which can effectively control harmful algae by mixing anthraquinone-based compounds derived from natural products.

Thus, the present inventors confirmed that the composition containing two or more of the anthraquinone-based compounds derived from natural products as an active ingredient has a marked synergistic effect on the control of harmful algae compared to the case of using one anthraquinone-based compound alone The present invention has been completed. Particularly, among the plant extracts containing anthraquinone compounds, fractions containing high one or more of chrysopanol, emodine, fishion, lane, and aloe-emodine are mixed to form chrysopanol, emidene, fishion, lane And it contains a synergistic effect on the control of harmful algae when it contains two or more of aloe-emodine, which results in improving the availability of natural products.

In accordance with the above object, in the present invention, two or more kinds of anthraquinone compounds derived from natural products, more preferably, two or more kinds of chrysopanol, emodine, fish, lane and aloe-emodine as active ingredients Provided is a composition for controlling harmful algae, and more particularly, provides a composition for controlling harmful algae containing chrysopanol, emodin, fish or a mixture thereof as an active ingredient.

According to the other object, the present invention provides a method for controlling harmful algae comprising treating the harmful algae composition in water or soil.

The present invention is described in more detail below.

The composition for controlling harmful algae according to the present invention contains two or more kinds selected from the anthraquinone compounds of the formula (1) as active ingredients.

[Formula 1]

[In Formula 1, R ₁ is H, R ₂ is selected from H, OH or OCH ₃ , R ₃ is selected from CH _3, COOH or CH ₂ OH.]

The anthraquinone compounds are chrysphanol (1,8-dihydroxy-6-methylanthraquinone), emodin (1,3,8-trihydroxy-6-methylanthraquinone), lanes as shown in the following table. [rhein (1,8-dihydroxy-6-carboxyanthraquinone)], fishion [physcion (1,8-dihydroxy-3-methoxy-6-methylanthraquinone)], aloe-emodin (1,8-dihydroxy -6- (hydroxymethyl) anthraquinone)].

The anthraquinone compounds of the present invention may be extracts or concentrates thereof which are directly synthesized or extracted from plants, more preferably Rumex, Rheum, Rhamnus, Polygonum ), Achyranthes, Aloe, Ardisia, Ant, Aster, Chassis, Chrysanthemum, Eupatorium, Evodia , Extracts from plants selected from Ligusticum, Paulownia, Petasites, Mentha, Inula, Hemerocallis, and mixtures thereof. Can be.

In the prymouth , Rumex crispus , Rumex japonicus , Rumex aquaticus , Rumex longifolius , Rumex conglomeratus , Rumex maritimus Rumex obtusifolius , Rumex gmelini , Rumex acetosa , Rumex acetosella are preferred to use the roots. Rheum undulatum , Rheum coreanum , Rheum palmatum , Rheum emodi , Rheum officinale , Rheum hotaoense , Rheum hotaoense , Rheum hotaoense Rheum nanum , Rheum glabricaule , Rheum ribes , Rheum spiciforme , Rheum tanguticum , or Rheum wittrochii It is preferable to use roots. It is preferable to use the roots of herbaceous persimmons , the main bones , and the mud-dangling as the herbaceous genus , and the seeds of the cassia mimosoides and the cassia tora . Other plants include Achyranthes bidentata , Ardisia japonica , Aster tataricus , Eupatorium spp, Aloe spp, Polygonum spp, and Sorghum tree ( Evodia officinalis ) , Hemerocallis fulva , Inula helenium , Mint ( Mentha haplocalyx ), Paulownia coreana , Butterbur ( Petasites japonicus ), Rhamnus davurica , Rhamnus crenata , Wormwood, It is preferable to use whole plants of chrysanthemum.

The plant extract according to the present invention may be prepared by extracting whole plants, plant roots, plant seeds or dried products thereof with a solvent, preferably by extracting the root of the larvae, rhubarb root or dried products thereof with a solvent. have. As the solvent, (C1-C5) alcohol or (C1-C5) alcohol aqueous solution, hexane, ethyl acetate, acetone, methylene chloride, chloroform, ethyl ether and the like may be used alone or in combination.

Extraction can be carried out using a conventional method, for example, by adding about 5 to 20 L of (C1 to C5) alcohol to 1 kg of dried plant roots and leaching at 20 to 60 ℃ for 6 hours to 20 days After obtaining, it can be further extracted with the other organic solvents exemplified above to obtain an organic solvent extract. In addition, the plant extract may be performed a fractionation process commonly used in the art, such as column chromatography. At this time, the column that can be used for the column chromatography is a silica gel column, reverse phase silica gel column or Sephadex column, and the eluent is an organic solvent such as hexane, ethyl acetate, chloroform, methylene chloride, acetone, C _1-5 alcohol, etc. Or these mixed liquid can be used.

In a preferred embodiment of the present invention, the anthraquinone-based compound may be obtained from the extract extracted with a solvent of a plant selected from the root of Rumex (Rumex), the root of Rheum (Rheum), the hexane fraction and It is more preferable to use a mixture of ethyl acetate fractions because of the synergistic action against pest control.

The hexane fraction is a step of preparing an alcohol extract by extracting a plant selected from the roots of the above-mentioned plants, more preferably roots of Rumex, roots of Rheum with (C1 ~ C5) alcohol , And the alcohol extract can be prepared by dissolving in (C1 ~ C5) alcohol or (C1 ~ C5) alcohol aqueous solution and extracting it with hexane to obtain a hexane extract.

The ethyl acetate fraction is prepared by extracting a plant selected from the roots of the above-mentioned plants, more preferably roots of Rumex, roots of Rheum with (C1 ~ C5) alcohol to prepare an alcohol extract. Step, dissolving the alcohol extract in (C1 ~ C5) alcohol or (C1 ~ C5) alcohol aqueous solution and separating it into a hexane layer, and (C1 ~ C5) alcohol or (C1 ~ C5) alcohol aqueous solution layer, and the Extracting the aqueous solution of (C1 ~ C5) alcohol or (C1 ~ C5) alcohol with ethyl acetate may be prepared through the step of obtaining an ethyl acetate extract.

The (C1 ~ C5) alcohol is more preferably used methanol or ethanol, concentrated by extracting the extract obtained by extraction with hexane or ethyl acetate under reduced pressure, and the concentrate is repeated silica gel column chromatography and Sephadex LH-20 column Purification by chromatography may be used to purely separate the anthraquinone compound.

Referring to the mixing ratio of the anthraquinone-based compound contained in the composition for controlling harmful algae according to the present invention, any one of (A) and aloe-emodin of chrysopanol, emodine, fish, lane and aloe The mixing ratio of the sum (B) of the other substances to be mixed can be adjusted in the range of 0.5 to 99.5%: 99.5 to 0.5% by weight ratio (A: B). If any one of the mixing weight ratio is less than 0.5% or more than 99.5%, the synergy effect due to mixing may be insignificant, and therefore, mixing in the above range is preferable.

When the hexane fraction and the ethyl acetate fraction are mixed, the amount may be adjusted to 0.5 to 99.5%, depending on the content of the anthraquinone compound in each fraction. If any one of the mixing weight ratio is less than 0.5% or more than 99.5%, the synergy effect due to mixing may be insignificant, and therefore, mixing in the above range is preferable.

The composition for controlling harmful algae containing two or more of the anthraquinone-based compounds derived from natural products according to the present invention as an active ingredient exhibits a selective control effect on harmful algae. The harmful algae include Microcystis , Anabaena , Oscillatoria , Oscillatoria , Aphanizomenon , Nodularia , Cochlodinium algae, and the like. . In particular, it exhibits a very high control against cyanobacteria such as Microcystis spp., Anabaena spp., Etc. , and is not harmful Chlorella spp., Senedesmus spp. Green algae such as) show a relatively low control effect. The selective control effect of the compounds on algae varies slightly from compound to compound. For example, chrysophanol inhibits microcistis tenfold more strongly than lanes, but has weaker activity than lanes for Cedencemus. Therefore, the control effect can be improved by varying the combination depending on the species to be controlled.

The mixed composition for controlling harmful algae of the present invention is a pharmaceutically acceptable solid carrier and liquid carrier for stable expression of the effect, enhancement of adhesion to the organism to be applied, and simplicity of transportation and treatment within the range of not inhibiting the effect of the mixture. , Surfactants such as liquid diluents, liquefied gas diluents, solid diluents, or other suitable auxiliaries such as emulsifiers, dispersants or foaming agents.

The control composition of the present invention may preferably be formulated in emulsion, hydrated, granulated, powdered, capsule and gelled formulations, and is preferably provided as a contact through a formulation such as a donut for buoyancy of the formulation. .

The present invention provides a harmful algae control method for controlling harmful algae by treating the composition for harmful algae according to the present invention in water or soil in which harmful algae as illustrated above. At this time, it is preferable to block the mass growth in advance by treating the composition in the early stage of the generation of harmful algae.

The harmful algae control method according to the present invention may have a low harmful algae control effect when the concentration of the active ingredient is too low when treated in the water or soil where the harmful algae is generated, and is uneconomical when the concentration of the active ingredient is too high. Since it may adversely affect the ecosystem, it is preferable to treat the harmful algae composition to a concentration of 0.01 μg / ml to 100 μg / ml based on the active ingredient, and the weight of the mixture of the hexane fraction and the ethyl acetate fraction In the case of the reference, it is preferable to treat the algae in water or soil to a concentration of 1 μg / ml to 100 μg / ml, which is appropriate in consideration of the type of harmful algae, the density of growth, the degree of growth, and the water environment. I can regulate it.

As described above, the mixture of the present invention uses an anthraquinone-based compound derived from a natural product in a pure state or a fraction of a plant extract, which controls and mixes harmful algae such as cyanobacteria relatively well while being less toxic to beneficial algae. Depending on the combination, the algae spectrum can also be changed and the activity is increased in the mixed treatment rather than in each single treatment. Therefore, it is possible to reduce the amount of side effects to the environment is possible to reduce the amount more, and the economic efficiency will also be good because the excellent algae activity is maintained. In addition, it is possible to use a mixture of extract fractions to increase the utilization efficiency of the extract sample. Therefore, the mixture of the present invention can be usefully used as a harmful algae control agent which is eco-friendly and excellent in efficacy.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

Example 1 Fraction layer mixing treatment of extract

(1) Preparation of Plant Extract Fraction

Rumex crispus roots collected outdoors were cut into 0.2 cm or less, immersed in methanol, left at room temperature for 5 days, filtered and concentrated under reduced pressure to obtain a methanol extract. It was dissolved in 70% methanol, extracted twice with the same amount of hexane, and the hexane layer and the aqueous layer were concentrated under reduced pressure. The concentrated aqueous layer was dissolved in distilled water and extracted twice with the same amount of ethyl acetate. The anthraquinone-based compound content was examined by the HPLC analysis method previously established with the hexane layer and the ethyl acetate layer thus obtained, and the content of chrysopanol, emodine, and pion in the hexane layer was 195, 17, 34 ㎍, respectively. It was / mg, ethyl acetate layer was 30.5, 68, 18.3 ㎍ / mg, respectively, hexane layer was chrysopanol, ethyl acetate layer was relatively high in the emodine. The column used for HPLC analysis was C18 (2) Phonomenex Luna (5 μm, 150 mm × 4.6 mm). The mobile phase was a solution containing 0.1% TFA in 75% methanol (volume) and the flow rate was 1 mL per minute. The extract sample was dissolved in methanol at a level of 1 mg / mL, and then injected into 10 μl and detected at 254 nm.

(2) Confirmation of control effect on seaweed

Hexane extract and ethyl acetate extract prepared in the above prepared in the final concentration of 0.0 ~ 0.6 ㎍ / ㎖, 0.0 ~ 2.5 ㎍ / ㎖, respectively, or a mixture of various mixtures after the growth inhibitory activity against blue algae It was confirmed by the method.

As a harmful alga, microcystis aeruginosa UTEX 2388, a cyanobacteria distributed by the Korea Institute of Bioscience and Biotechnology, was used.

First, microcistis aeruginosa incubated in culture medium (Allen medium) was inoculated, and the initial concentration of algae at this time was about 0.06 at 670 nm using a microplate reader. Was made. 10 ml of the solution was injected into a 50 ml culture bottle, and 100 µl of the test solution and the control solution prepared as follows were administered to the strain culture. For test solution preparation, each methanol extract was dissolved in dimethylsulfoxide containing 100 μg / ml of Tween 20 ™ and then prepared at various concentrations using the same solvent. As a result, the final concentrations of dimethylsulfoxide and Tween 20 in the culture were about 1% and 2 μg / ml, respectively. On the other hand, a solution in which the solvent was added to distilled water under the same conditions was used as a control. The algal cultures to which these test agents were administered were then incubated at 25 ° C., 14 hour photoperiod, 40-60 μmol ⁻² s ^−1, and 100 rpm. The absorbance was measured 6 days after treatment of the test solution, and was investigated through the absorbance-building correlation (Kim et al., Aquatic Botany 85: 1-6, 2006) (Equation 1). Afterwards, the algal control effect of the composition for the control was calculated as the degree of inhibition of dry matter in the control group (%).

[Equation 1]

Y = -0.00837X ² + 0.2524X + 0.00383

In the above formula, Y represents absorbance and X represents dry weight.

The interaction between the two mixtures was evaluated by Colby SR. Weeds 15: 20-22, 1967. If the actual control value (observed value) indicated by the mixture treatment is greater than the expected value (expected by Equation 2, expected value), it indicates synergism, equality, and antagonism action will be.

[Equation 2]

E = (X + Y)-XY / 100

Where X is% inhibition at p concentration of Compound A and Y is% inhibition at q concentration of Compound B.

As a result of the experiment, as shown in FIG. 1, the observed value was higher than expected in the hexane layer 0.1-0.3 ㎍ / ml + ethyl acetate 0.3-1.25 ㎍ / ml in each concentration combination, showing a strong synergy. To completely control the microcistis, a hexane layer 0.15 μg / ml + ethyl acetate 1.25 μg / ml, or a hexane layer 0.3 μg / ml + ethyl acetate 0.6 μg / ml was sufficient. However, in the case of alone, the hexane layer and the ethyl acetate layer had to be treated at concentrations of 0.6 and 2.5 ㎍ / ml, respectively.

We tried to see if this result could actually produce beneficial results in practical terms. That is, the yield of the hexane layer and the ethyl acetate layer obtained by the above method from the ethanol extract of the root of Rheum platanum , a rhubarb plant, was about 1-2% and about 20%, respectively. In general, since the activity of the hexane layer is relatively higher than that of the ethyl acetate layer, only the hexane layer will be used. However, when the two fractions were mixed and treated at a concentration capable of controlling 95% of the microcistises, the calculated volume was more than twice as high as that of the hexane layer alone.

Example 2 Mixing Treatment of Chrysopanol and Emodine

When the mixed treatment as in Example 1 in the extract as well as the purely separated components of the extract, it was examined whether there is a synergistic effect in the algae action.

(1) securing the compound

Chrysophanol (purity 98%) and emodine (purity 90% or more) were all purchased from Sigma-Aldrich.

(2) Confirmation of control effect on seaweed

Chrysophanol + emodine was prepared in the range of 0.125 ~ 0.5 ㎍ / ㎖ + 0.25 ~ 1.0 ㎍ / ㎖ in the final concentration, respectively, and the growth inhibitory activity against cyanobacteria was confirmed. All other experiments were performed as in Example 1 and the interaction between the two mixtures was evaluated by Colby method.

As a result of the experiment, as shown in Table 1, the measured value was higher than expected at various concentration combinations, which showed synergy. In each concentration combination, the excellent combination that completely controlled the microcistis at the present experimental conditions was 0.125 µg / ml of chrysophanol + 0.25-0.75 µg / ml of emodin.

On the other hand, except that the elapsed days after the treatment composition treatment was changed from 6 days to 9 days, another experiment was carried out under the same experimental conditions and the result of the investigation showed synergy as shown in FIG.

TABLE 1

Example 3 Mixing Treatment of Emodine and Fishion

When not only the extract but also the pure separation component of the extract as in Example 2, the mixture was examined whether there is a synergistic effect in the algae action.

(1) securing the compound

Emodine (purity 90% or more) purchased from Sigma-Aldrich company and Pion (purity 90% or more) purely separated from the larvae were used.

(2) Confirmation of control effect on seaweed

Emodine + fish was prepared in the range of 0.1 ~ 0.8 ㎍ / ㎖ + 0.15 ~ 0.6 ㎍ / ㎖, respectively, at a final concentration, and after 6 days, growth inhibition activity against cyanobacteria was confirmed. . All other experiments were performed as in Example 1 and the interaction between the two mixtures was evaluated by Colby method.

As a result, as shown in Table 2, the measured value was higher than expected in the combination of 0.2 μg / ml of emodin + 0.3 μg / ml of psion and showed synergy. At other concentrations, however, weak synergy was observed.

TABLE 2

Example 4 Mixing Treatment of Chrysophanol and Fishion

When not only the extract but also the pure separation component of the extract as in Example 2, the mixture was examined whether there is a synergistic effect in the algae action.

(1) securing the compound

Chrysophanol (purity 98%) purchased from Sigma-Aldrich company and fish pion (purity 90% or more) purely separated from the larvae were used.

(2) Confirmation of control effect on seaweed

Chrysophanol + fish was prepared in the final concentration range of 0.05 to 0.2 μg / ml + 0.15 to 0.6 μg / ml, respectively, or a combination of various mixtures, and growth inhibitory activity against cyanobacteria was confirmed. All other experiments were performed as in Example 2 and the interaction between the two mixtures was evaluated by Colby method.

As a result, as shown in FIG. 3, the measured value was higher than expected at various concentration combinations, indicating good synergy. The combination which showed the best activity under these experimental conditions was found in the treatment condition of 0.05 μg / ml of chrysophanol + 0.3 μg / ml of fish.

Example 5 Mixing Treatment of Emodine and Lane

When not only the extract but also the pure separation component of the extract as in Example 2, the mixture was examined whether there is a synergistic effect in the algae action.

(1) securing the compound

Emodine (purity 90% or more) and lane (purity 98%) purchased from Sigma-Aldrich Company were used.

(2) Confirmation of control effect on seaweed

Emodine + lanes were prepared in the range of 0.1 ~ 0.8 ㎍ / ㎖ + 0.125 ~ 1.0 ㎍ / ㎖ in the final concentration, respectively, and then the growth inhibitory activity against cyanobacteria was confirmed. All other experiments were performed as in Example 2 and the interaction between the two mixtures was evaluated by Colby method.

As a result, as shown in FIG. 4, the measured value was higher than expected at various concentration combinations, indicating a good synergy. The combination showing the best synergistic activity under these experimental conditions was found in the treatment conditions of 0.4 ~ 0.5 ㎍ / ㎖ Emodine + 0.25 ~ 0.5 ㎍ / ㎖ concentration.

1 is a graph showing that the mixture of the hexane fraction layer and the ethyl acetate fraction layer of the extract of Rinja root has a synergistic effect on the control of microcistis aeruginosa,

FIG. 2 is a graph showing that the mixture of chrysopanol and emodine is synergistic with respect to microcystis aeruginosa control,

3 is a graph showing that the mixture of chrysopanol and fish cation has a synergistic effect on microcystis aeruginosa control,

4 is a graph showing that the mixture of emodin and lanes is synergistic with respect to microcistis aeruginosa control.

Claims (12)

As active ingredients, chrysophanol and ememodine; Emodin and fish; Chrysopanol and fish; And anthraquinone-based compounds selected from the group consisting of emodine and lanes, wherein the weight ratio of chrysophanol: emodine is 1: 1 to 6, the weight ratio of emodine: fishy: 1: 1.5-3 It contains a weight ratio of 1 to 4: 3 to 12, the weight ratio of emodine: lane 1: 1: 2525 to 1.25, composition for controlling harmful algae. delete The method of claim 1, The anthraquinone-based compound may include roots of the genus Rumex, roots of the Rheum, roots of Ligusticum, and fruit of the genus Chassis; Achyranthes bidentata , Ardisia japonica , Aster tataricus , Eupatorium spp, Aloe spp, Polygonum spp, Evodia officinalis , Royal trout Hemerocallis fulva , Inula helenium , Mint ( Mentha haplocalyx ), Paulownia coreana , Butterbur ( Petasites japonicus ), Rhamnus davurica , Rhamnus crenata , Wormwood, Chrysanthemum and mixtures thereof A composition for controlling harmful algae, characterized in that obtained from an extract prepared by extracting a plant from a solvent with a solvent or a concentrate thereof. The method of claim 3, wherein The solvent is a composition for controlling harmful algae, characterized in that selected from water, (C1-C5) alcohol, hexane, ethyl acetate, acetone, methylene chloride, chloroform, ethyl ether or a mixture thereof. The method of claim 4, wherein The anthraquinone-based compound may include roots of the genus Rumex, roots of the Rheum, roots of Ligusticum, and fruit of the genus Chassis; Achyranthes bidentata , Ardisia japonica , Aster tataricus , Eupatorium spp, Aloe spp, Polygonum spp, Evodia officinalis , Royal trout Hemerocallis fulva , Inula helenium , Mint ( Mentha haplocalyx ), Paulownia coreana , Butterbur ( Petasites japonicus ), Rhamnus davurica , Rhamnus crenata , Wormwood, Chrysanthemum and mixtures thereof Composition for controlling harmful algae, characterized in that obtained from the hexane fraction and ethyl acetate fraction of the plant selected from the group consisting of. The method of claim 5, The hexane fraction is the root of the genus (Rumex), the root of the genus Rheum, the root of Ligusticum (Ligusticum), the fruit of the tea (Cassia); Achyranthes bidentata , Ardisia japonica , Aster tataricus , Eupatorium spp, Aloe spp, Polygonum spp, Evodia officinalis , Royal trout Hemerocallis fulva , Inula helenium , Mint ( Mentha haplocalyx ), Paulownia coreana , Butterbur ( Petasites japonicus ), Rhamnus davurica , Rhamnus crenata , Wormwood, Chrysanthemum and mixtures thereof Preparing an alcohol extract by extracting a plant selected from the group consisting of (C1-C5) alcohol; And Dissolving the alcohol extract in (C1 ~ C5) alcohol or (C1 ~ C5) alcohol aqueous solution and extracting it with hexane to obtain hexane extract; A composition for controlling harmful algae, which is prepared from. The method of claim 5, The ethyl acetate fraction is the root of the genus (Rumex), the root of the genus Rheum, the root of the Ligusticum (Ligusticum), the fruit of the genus Chassis (Cassia); Achyranthes bidentata , Ardisia japonica , Aster tataricus , Eupatorium spp, Aloe spp, Polygonum spp, Evodia officinalis , Royal trout Hemerocallis fulva , Inula helenium , Mint ( Mentha haplocalyx ), Paulownia coreana , Butterbur ( Petasites japonicus ), Rhamnus davurica , Rhamnus crenata , Wormwood, Chrysanthemum and mixtures thereof Preparing an alcohol extract by extracting a plant selected from the group consisting of (C1-C5) alcohol; Dissolving the alcohol extract in (C1 ~ C5) alcohol or (C1 ~ C5) alcohol aqueous solution and separating it into a hexane layer, and (C1 ~ C5) alcohol or (C1 ~ C5) alcohol aqueous solution layer; And Extracting the (C1 ~ C5) alcohol or (C1 ~ C5) alcohol aqueous layer with ethyl acetate to obtain an ethyl acetate extract; Harmful algae control composition, characterized in that it was prepared through. delete delete The method of claim 1 Harmful birds are selected from the group consisting of algae of the genus Microcystis , Anabaena , Oscillatoria , Aphanizomenon , Nodularia and Cochlodinium Characterized in that, harmful algae control composition. Of claim 1, claim 3 to claim 7, and claim 10, wherein any one of the harmful if algae micro-a for controlling composition seutiseu (Microcystis), Ana vena (Anabaena), come la thoria (Oscillatoria) wherein Aphanizomenon Menon ( Aphanizomenon ) , Nodularia or Cochlodinium ( Cochlodinium ), including the treatment of harmful algae, including the treatment of water or soil inhabiting algae. The method of claim 11, The harmful control composition is characterized in that the treatment to the concentration of 0.01 μg / ㎖ to 100 ㎍ / ㎖ in water or soil based on the concentration of the active ingredient, harmful algae control method.

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