KR101013946B1 - Composition comprising menadione based compound for enhansing control of harmful algae, and method for controlling harmful algae using the same - Google Patents

Abstract

The present invention is a compound selected from menadione, menadione sodium bisulphite or juglone, and at least one anthraquinone compound or a plant extract containing the same as an active ingredient, preferably The present invention relates to a mixed composition for controlling harmful algae prepared by mixing a plant extract containing high chrysopanol, and a method for controlling harmful algae using the same, wherein the mixed composition of the present invention is selective for harmful algae such as cyanobacteria and red algae. Since the growth inhibitory effect, the control composition containing the same can be usefully used as a harmful algae control agent having excellent environmental friendliness.

Harmful algae, cyanobacteria, chrysopanol, menadione, plant extracts

Description

COMPOSITION COMPRISING MENADIONE BASED COMPOUND FOR ENHANSING CONTROL OF HARMFUL ALGAE, AND METHOD FOR CONTROLLING HARMFUL ALGAE USING THE SAME}

The present invention provides a plant extract comprising a compound selected from menadione, menadione sodium bisulfite, or juglon and at least one anthraquinone compound or a plant extract containing the same as an active ingredient, preferably a chrysopanol The present invention relates to a mixed composition for controlling harmful algae prepared by mixing and a method for controlling harmful algae using the same.

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 habitats of harmful algae Oscillatoria perornata -flavor) (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. Among the compounds used, many compounds are preferred to be replaced in the future due to their relative importance in terms of cost and control effects rather than consideration of phosphorus toxicity or environmental stability. For example, copper sulfate, chlorine, alkaline earth metal peroxides are fast-acting, 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, 2003). In the case of chlorine, it is possible to work with organic substances (humic acid, fulvic acid, etc.) in water to increase the possibility of carcinogenic trihalomethanes. 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), Piserelin A and B (fischerellin A and B, Hagmann et al . , Tetrahedron Lett. 37: 6539-6542, 1996; Papke et al . , Tetrahedron Lett. 38: 379-382, 1997), oxygenated fatty acids and lysine Potassium ricinoleate, Duke et al . , Weed Sci. 50: 138-151, 2002), L-2-azetidinecarboxylic acid (Kim et al., Aquatic Botany 85: 1-6, 2006), rutacridone Rutacridone epoxide (Meepagala et al., Phytochemistry 66: 2689-2695, 2005), Ethyl 2-methylacetoacetate (Li & Hu, Appl. Environ. Microbiol. 71: 6545-6 553, 2005), and others (Duke et al . , Weed Sci. 50: 138-151, 2002; Schrader et al., Aquaculture 163: 85-99, 1998) and are known as botanical barley straw and rye grass ( 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, and they are different depending on the compound but need to be compensated for practical use, that is, they have low activity or narrow working spectrum 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 technology for developing and applying a mixture having synergy with each other is required.

On the other hand, Menadione and Zerglon have been reported to have growth inhibitory activity against microorganisms or phytoplankton (WO 2001 / 028328A1; US Patent 2002 / 0117079A1; Wright et al., Water Research 41: 1294-1302, 2007; Wright; Et al. , Environ. Technol. 28 (3): 309-320, 2007; Kessler CT, J. Chem. Ecol. 15: 2127-2134, 1989), or natural products that, when mixed with them, may exhibit significant synergistic effects. There have been no reports on the investigation of natural compounds and their application to control harmful algae as a mixture.

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, an object of the present invention is to mix a compound derived from natural products with a compound selected from menadione, menadione sodium bisulfite or juglon to bring about a significant increase in the harmful algae control effect and a new composition for harmful algae control It is to provide a harmful algae control method using the same.

Therefore, the inventors of the present invention, when a specific plant extract or an active ingredient is added to a compound selected from menadione known as vitamin K3, menadione sodium bisulfite synthesized to increase the water solubility of menadione, or juglon, to control harmful algae The present invention was completed by confirming that there is a synergistic effect.

In accordance with the above object, in the present invention, a plant extract containing at least one anthraquinone-based compound or an active ingredient thereof is added to a compound selected from menadione, menadione sodium bisulfite, or zerglon to raise harmful birds. It provides a composition for controlling harmful algae having an action.

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 present invention provides a compound selected from (A) menadione, menadione sodium bisulphite or juglone; And

(B) at least one anthraquinone compound of Formula 1 or a plant extract comprising the same;

It provides a composition for controlling harmful algae containing.

[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.]

Among the anthraquinone compounds of the present invention, preferred are chrysophanol (1,8-dihydroxy-6-methylanthraquinone) and emodin (1,3,8-trihydroxy-6-methylanthraquinone) of Chemical Formula 1. ], Lanes [rhein (1,8-dihydroxy-6-carboxyanthraquinone)], fishes [physcion (1,8-dihydroxy-3-methoxy-6-methylanthraquinone)], aloe-emodin (1, 8-dihydroxy-6- (hydroxymethyl) anthraquinone)] or mixtures thereof.

The anthraquinone compounds of the present invention can be used for organic synthesis or separated from plants. The plants include Rumex, Rheum, Rhamnus, Rhamnus, Polygonum, Achyranthes, Aloe, Aloe, Ardisia, Ant (Aster), Cassia, Chrysanthemum, Eupatorium, Evodia, Ligusticum, Paulownia, Paulownia, Petetasites, Mentha ), A plant belonging to the genus Inula, Hemerocallis, and a mixture thereof. 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 , Mugwort, Chrysanthemum It is preferable to use whole plants of.

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, water, (C1-C5) alcohol, hexane, ethyl acetate, acetone, methylene chloride, chloroform, ethyl ether and the like can 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 plant extract is the root of the genus (Rumex), the root of Rheum (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 Extracting a plant selected from the group consisting of (C1 ~ C5) alcohol to prepare an alcohol extract, dissolving the alcohol extract in (C1 ~ C5) alcohol or (C1 ~ C5) alcohol aqueous solution and hexane, ethyl acetate Or hexane fraction prepared by extracting with methylene chloride, and concentrating the obtained hexane extract, ethyl acetate extract or methylene chloride extract under reduced pressure. Fraction or methylene chloride fraction.

In a more preferred embodiment, the anthraquinone-based compound of Formula 1 is a hexane, ethyl acetate or hexane using an extract extracted from the root of Rumex (Rumex), the root of Rheum (Rheum) extract with methanol or ethanol After obtaining a methylene chloride extract and concentrated under reduced pressure, the concentrate was purified and separated by repeated silica gel column chromatography and Sephadex LH-20 column chromatography.

An anthraquinone compound of Formula 1 or a plant extract containing the same as an active ingredient according to the present invention, a mixed composition of menadione, menadione sodium bisulfite or juglon has 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.

In mixing Compound B (at least one of the anthraquinone compounds) with Compound A (one of Menadione, Menadione Sodium Bisulfate, and Zerglon), the weight ratio of Compound A to Compound B active ingredient is 1 to 99%: It may be mixed in the range of 99% to 1%, more preferably 5 to 95%: 95 to 5%. If any one of the above mixed weight ratios is less than 1% or more than 99%, the synergistic effect of the mixing may be insignificant, so it is preferable to mix in the above range. In the case of a mixture of menadione and chrysopanol, in order to obtain a better effect, 60 to 95% by weight of menadione, 5 to 40% by weight of chrysophanol, or 5 to 30% by weight of menadione and 70 to chrysopanol It is preferred to mix at 95% by weight, and even more preferably at 70 to 95% by weight of menadione and 5 to 30% by weight of chrysophanol.

In the case of containing a plant extract containing at least one of the anthraquinone compounds, it is preferable to consider the anthraquinone compounds in the plant extract. The content of the anthraquinone-based compound contained in the plant extract may vary depending on the preparation method of the plant extract, so there is no need to limit the content, but it has a range of 0.01 to 99.99 wt% based on the total weight of the plant extract.

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 the adverse effects on the ecological environment may appear, it is preferable to treat the harmful algae control composition to a concentration of 0.01 μg / ml to 100 μg / ml based on the active ingredient, which includes the types of harmful algae, density of occurrence, The growth can be appropriately adjusted in consideration of the quality of the environment and the water environment.

As described above, the mixture of the present invention is relatively toxic to the beneficial algae, while controlling the harmful algae, such as southern algae relatively well and the activity is increased in the mixed treatment rather than 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. Therefore, the mixture of the present invention can be usefully used as a pest control agent that is natural 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 Mixed Composition of Menadione and Tertiary Hexane Extract

Menadione is a vitamin K3 compound known to have microbial control effects and has been reported to have control activity against phytoplankton (Wright DA, Water Research 41: 1294-1302, 2007). Searching for other compounds that are synergistic with menadione is important for practical research because it can reduce chemical doses and improve efficacy. In this experiment, the interaction of the algae activity by mixing the hexane extract fraction layer was investigated.

(1) Preparation of Hexane 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 was concentrated under reduced pressure. As a result of investigating the content of the hexane layer obtained by the previously established HPLC analysis method, chrysophanol and emodine were relatively high among the anthraquinone compounds. 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) control effect of the control composition

The control effect of the hexane fraction layer of the plant extract prepared above and menadione (purity 99%, manufactured by Sigma Aldrich) or menadione sodium bisulfite (purity 95%, manufactured by Sigma Aldrich) was controlled. Evaluated.

Specifically, the hexane fractionation layer has a final concentration of 0.0-0.6 μg / ml and menadione or menadione sodium barsulphite in a final concentration of 0.05-0.2 μg / ml. Growth inhibition activity was confirmed.

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. 6 days after treatment of the test solution, the absorbance was measured and investigated through the absorbance-building correlation equation (Kim et al., Aquatic Botany 85: 1-6, 2006) (Equation 1). , 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

[Equation 1, Y represents the absorbance, X represents the building.]

[Equation 2]

E = (X + Y)-XY / 100

[In Formula 2, X represents% inhibition at p concentration of Compound A, and Y represents% inhibition at q concentration of Compound B.]

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.

As a result of the experiment, as shown in FIG. 1, the measured value was higher than expected in the menidione 0.05 ~ 0.15 ㎍ / ㎖ + hexane fraction of 0.1 ~ 0.5 ㎍ / ㎖ of each concentration combination showed a strong synergistic effect, this experimental condition In order to completely control the microcistis at, treatment with Menadione 0.05 μg / ml + Hexane fraction layer 0.3 μg / ml, or Menadione 0.1 μg / ml + Hexane fraction layer 0.15 μg / ml was sufficient. However, in the case of alone, it was possible to treat the menadione and hexane fractionation layers at concentrations of 0.6 and 0.2 μg / ml.

On the other hand, experiments with menadione sodium bisulfite (water-soluble menadione) showed a slightly lower effect than menadione, but showed the same tendency in synergy (data not shown).

Example 2 Mixed Composition of Menadione and Rhubarb Plant Extracts

In this experiment, we investigated the interaction of algae activity with the mixing of Rheum undulatum extract produced in China with menadione.

(1) Securing Rhubarb Plant Extracts and Menadione

Plant extracts were purchased from a Chinese company (Shaanxi Jiahe Phytochem Co., LTD), an ethanol extract sample (containing 57.9% of emodin) from Rheum palmarum dried roots (hereinafter referred to as Rp-K6). Dion used Sigma-Aldrich company (99% purity).

(2) Confirmation of control effect on seaweed

Menadione and Rp-K6 were prepared in the final concentration of 0.05 ~ 0.2 ㎍ / ㎖, 0.126 ~ 1.0 ㎍ / ㎖, respectively, or a mixture of various mixtures, 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 shown in FIG. 2, it was shown that the measured value was higher than expected at menidione 0.05 to 0.15 ㎍ / ml + Rp-K6 0.125 to 0.5 ㎍ / ml in each concentration combination, showing a strong synergy. To completely control the microcistis at, treatment with 0.05 g / ml of menadione + 0.5 g / ml of Rp-K6, or 0.1 g / ml of menadione + 0.125 g / ml of Rp-K6 was sufficient. However, it was possible to treat menadione and Rp-K6 at concentrations of 0.2 and 1.0 or higher (µg / ml) alone.

Overall, higher synergy was observed in the extract mix than in the following single component mix (Example 3), presumably because other ingredients present in traces in the extract are more influential toward efficacy. This feature shows very desirable results showing that the use of moderately separated extracts (eg, hexane fractionation layers) rather than purely separated components is more advantageous in terms of efficacy and economics.

Example 3 Mixed Composition of Menadione and Anthraquinone Compound

In addition to the extract, the pure separation component of the extract, when mixed with menadione as in Example 1 and Example 2, was examined whether there is a synergistic effect in the algae action.

(1) securing the compound

Chrysophanol (purity 98%), emodin (purity 90% or more), and menadione (purity 99%) were all purchased from Sigma-Aldrich.

(2) Confirmation of control effect on seaweed

In the case of menadione + chrysopanol, the growth inhibitory activity against cyanobacteria was confirmed after preparing single or various mixture combinations in the ranges of 0.05 to 0.1 μg / ml + 0.05 to 0.2 μg / ml, respectively. In the case of menadione + emodine, the growth inhibitory activity against cyanobacteria was confirmed after the preparation of single or various mixtures in the range of 0.05 ~ 0.1 ㎍ / ㎖ + 0.125 ~ 1.0 ㎍ / ㎖ at the final concentration, respectively. 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 and Table 2, the measured value was higher than expected at various concentration combinations, which showed synergy. When mixed with menadione, chrysophanol had a relatively better synergy than emodidine. In each concentration combination, the superior combination that completely controls the microcistis at this experimental condition was 0.05 g / ml of menadione + 0.2 g / ml of chrysophanol, 0.1 g / ml of menidione and 0.1 g / ml of cresophanol, and menidione. 0.05 μg / ml + emodine 0.5 μg / ml.

TABLE 1

TABLE 2

(3) Control effect of cyanobacteria according to the mixing ratio in the same dose of chrysopanol + menadione mixture

The synergy was further confirmed, and the experiment was performed to determine the difference in the control effect according to the mixing ratio of each compound in the same dose of the mixture. That is, the mixing ratio of the chrysophanol (A) and the menadione (B) compounds to the weight ratio is 0:10, 2: 8, 4: 6, 5: 5, 6: 4, 8: 2, and 10: 0, respectively. The prepared sample was dissolved in a solvent as in Example 1 to prepare test compound solutions of various concentrations. This was diluted in an allene medium containing cyanobacteria (Microcistis aeruginosa) to investigate the growth inhibitory activity of the test solution at 0.015-1.0 ㎍ / ㎖ concentration conditions, all other experimental conditions were the same as in Example 1. The synergistic effect of the mixing ratio was evaluated by first obtaining the actual 50% growth inhibition concentration (actual value EC50) in each sample, and then calculating the theoretical 50% growth inhibition concentration (theoretical value EC50) by using Equation 3 (Gisi). Et al. , Trans. Brit. Mycol. Soc. 85: 299-306, 1985). At this time, if the actual 50% growth inhibition concentration (acting EC50) of each sample with different mixing ratio is lower than the theoretical 50% growth inhibition concentration (theoretical EC50), synergism, if it is equal, the antagonism is larger It shows the action.

&Quot; (3) "

EC50 (Th) = (a + b) / [a / EC50 (A) + b / EC50 (B)]

Here, a and b represent the mixing ratios of Compounds A and B, and EC50 (A) and EC50 (B) represent 50% growth inhibitory concentrations when Compounds A and B were treated alone.

As a result, as shown in Table 3, the measured value was higher than the theoretical value at all concentration combinations, and it showed synergistic effect. Among these mixing ratios, the effect was very good. It was less than 40% and the next good effect was when the mixture ratio of menadione and chrysopanol was 20% and 80%.

[Table 3]

Example 4 Mixed Composition of Zerglon and Anthraquinone Compound

When mixing with an anthraquinone-based compound, it was examined whether synergistic effect appeared in the algae action as in Example 7 even when juggleon was used instead of menadione.

(1) securing the compound

Chrysophanol (purity 98%), emodin (purity 90% or more), and zerglon (99% purity) were all purchased from Sigma-Aldrich.

(2) Confirmation of control effect on seaweed

Zerglon, known to be contained in walnut tree immature bark, is a natural ingredient of naphthoquinone family that has a similar chemical structure to menadione, and its growth inhibition activity against cyanobacteria is stronger than menadione. In the case of Zeroglon + Chrysopanol, growth inhibitory activity against cyanobacteria was confirmed after preparing single or various mixtures in the range of 0.0125 to 0.05 μg / ml + 0.05 to 0.2 μg / ml, respectively. . In the case of zerglon + emodine, single or various mixture combinations were prepared in the range of 0.0125 to 0.05 μg / ml + 0.125 to 1.0 μg / ml, respectively. All other experiments were performed as in Example 4 and the interaction between the two mixtures was evaluated by Colby method.

As a result of the experiment, as shown in Table 4 and Table 5, the measured value was higher than expected at various concentration combinations, which showed synergy. In each concentration combination, the excellent combination that completely control the microcistis at this experimental condition was: Zerglon 0.0125 μg / ml + Chrysophanol 0.1 μg / ml, Zergon 0.025 μg / ml + Chrysophanol 0.05 μg / ml, Zerglon 0.025 μg / ml + emodin 0.25 μg / ml.

[Table 4]

TABLE 5

1 is a graph showing that the mixture of the hexane fractionation layer of menadione and the extract of Roots has a synergistic effect against microcistis aeruginosa control,

2 is a graph showing that a mixture of menadione and Rhubarb root extract (containing 55.9% of emodin) is synergistic with respect to microcystis aeruginosa control.

Claims (10)

(A) a compound selected from menadione, menadione sodium bisulphite or juglone; And (B) hexane fraction or rhubarb ethanol extract of anthraquinone-based compound selected from chrysophanol or emodin, or an extract of methanol containing same; Containing, and the weight ratio of menadione or menadione sodium bisulfite: chrysophanol is 1: 0.25-4, and the weight ratio of menadione or menadione sodium bisulfite: emodine is 1: 1.25-20, Gloss: Chrysophanol weight ratio of 1: 2 to 8, Zerglon: Emodine weight ratio of 1: 5 to 40, the composition for controlling harmful algae. delete delete delete delete 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. Claim when the 1 or claim 8 compositions for harmful algae control micro seutiseu (Microcystis), Ana vena (Anabaena), come la thoria (Oscillatoria), Aphanizomenon Menon (Aphanizomenon), no dyulra Liao (Nodularia) or co keulrodi titanium ( Cochlodinium ) A method for controlling harmful algae, characterized in that it is treated in the water or soil inhabiting the genus. The method of claim 9, Toxic algae control method, characterized in that for treating the harmful composition of the mixed composition to 0.01 μg / ㎖ to 100 ㎍ / ㎖ in water or soil on the basis of the active ingredient.

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