KR20220075053A - Composition of repellent containing plant oil and related chemical compounds for Controlling Haemaphysalis longicornis - Google Patents

Abstract

The present invention relates to a natural repellent composition for controlling small mites of Saturella montanya essential oil and Amorasia rusticana essential oil, and more specifically, Saturella montanya essential oil, ah It relates to a natural repellent composition comprising Morasia rusticana essential oil and their active ingredients or derivative compounds of active ingredients as an active ingredient.

Description

Repellent active composition for controlling small mites comprising plant essential oil and related compounds {Composition of repellent containing plant oil and related chemical compounds for Controlling Haemaphysalis longicornis}

The present invention relates to a composition for controlling small mites, and more particularly, having repellent activity against mites, Satureja montana ( Satureja montana ) essential oil, Amoracia rusticana ) Active ingredients of essential oil Or, it relates to a repellent composition comprising a derivative compound of the active ingredient.

Ticks are the first arthropods recognized as vectors of pathogens, and are currently recognized as major arthropod vectors that transmit zoonoses related diseases to humans and livestock along with mosquitoes. In particular, Haemophysalis longicorni ( Haemophysalis longicorni ), a member of the family Mites, is found throughout the country from May to August, and is known to be parasitic on animals that live outdoors.

Small Sophie tick is a major bloodsucking tick that transmits Sever Fever with Thrombocytopenia Symdrome Virus (SFTSV). In Korea, over the past five years, a total of about 1,000 patients infected with SFTSV and 180 deaths are threatening the lives of the people.

The fatality rate of SFTS patients in Korea is about 18% due to the small sophie tick, but there are no vaccines or antivirals that exist around the world, posing a threat to human survival. In addition, the development of new drugs such as vaccines or therapeutics that can treat patients infected with SFTSV takes a long time of more than 10 years including clinical trials, so realistically, we focus on the control of small sophia tick, a major vector of zoonotic diseases. It is imperative to match

DEET ( N,N -diethyl- m -toluamide) is a compound that protects humans and animals from mosquitoes/mite, and is known as a repellent with excellent repellent activity and durability. According to the US Centers for Disease Control and Prevention, approximately 30% of the US population uses DEET and DEET-based products as mosquito/mite repellents, and it is estimated that approximately 23-29% of children are exposed to DEET. DEET is effective as a repellent, but its use is limited because side effects have been reported for children, pregnant women, and fetuses. Therefore, it is expected that the development of repellents using plant-derived biomaterials will make a great contribution to public health, agro-food, and disease science fields in order to effectively control the small Sophia mite.

Patent Document 1: Korean Patent Publication KR 20090040269 A

The present inventors confirmed that Saturella montana essential oil and Amorasia rusticana essential oil exhibit a high repelling effect against small Sophie mite while searching for a method for controlling Sophie mite, and completed the present invention.

Accordingly, it is an object of the present invention to provide a repellent composition for controlling small mites comprising active ingredients of Saturella montana essential oil and Amorasia rusticana essential oil, and derivative compounds of these active ingredients.

In order to achieve the above object, the present invention relates to a natural repellent for small cattle mites, 5-isopropyl-2-methylphenol, an active ingredient of Saturella montanya essential oil, and 5-isopropyl-, a derivative thereof Provided is a repellent active composition comprising as an active ingredient at least one compound selected from 2-methylaniline, phenethyl isothiocyanate, an active ingredient of Amorasia rusticana essential oil, and benzyl isothiocyanate, a derivative compound thereof .

In addition, the present invention provides a tick repellent prepared using the repellent active composition.

Saturella montana essential oil, Amorasia rusticana essential oil, and their active ingredients and derivative compounds of active ingredients according to the present invention are plant-derived natural repellents for small ticks, and have excellent repelling activity against small ticks, It is effective as a simple, economical and eco-friendly formulation.

1 is a diagram showing a climbing test (climbing test) test carried out in the present invention.
2 shows the compounds (1) 5-isopropyl-2-methylphenol, (2) 5-isopropyl-2-methylaniline, (3) benzyl isothiocyanate and (4) phenethylisothio, which were carried out in the present invention. It is a graph showing repelling activity after 60 minutes according to 1:1 mixing of each compound of cyanate.

In order to achieve the above object, the present invention relates to a natural repellent for small cattle mites, 5-isopropyl-2-methylphenol, an active ingredient of Saturella montanya essential oil, and 5-isopropyl-, a derivative thereof Provided is a repellent active composition comprising as an active ingredient at least one compound selected from 2-methylaniline, phenethyl isothiocyanate, an active ingredient of Amorasia rusticana essential oil, and benzyl isothiocyanate, a derivative compound thereof .

In the present invention, "Saturella montanya" belongs to the family Lamiaceae, and is a perennial semi-shrub that grows wild in Europe, Russia, Turkey, and northern Africa. Saturella montana is a herb with a strong scent and has been used as a spice, tea, and seasoning since ancient times. “Amorasia rusticana” belongs to the cruciferous family, and is a perennial plant that grows widely in southeastern Europe and western Asia. The root of Amorasia rusticana is called mustard radish and is widely used as a spice for its spicy taste.

The Saturella montanya essential oil in the present invention can be used that is distributed and sold in the market, and the Amorasia rusticana essential oil is extracted by continuous steam distillation.

In the present invention, the '5-isopyrophyl-2-methylphenol' compound is represented by the formula (1), and the '5-isopropyl-2-methylaniline' is a compound represented by the following formula (2), and ' The 'phenethylisothiocyanate' compound is represented by the following formula (3), and the 'benzyl isothiocyanate' is a compound represented by the following formula (4).

<Formula 1>

<Formula 2>

<Formula 3>

<Formula 4>

The 5-isopyrrophyll-2-methylphenol compound of formula (1) and 5-isopropyl-2-methylaniline of the formula (2) may be those that are distributed and sold in the market, and, in addition, the formula (2) The phenethyl isothiocyanate compound and benzyl isothiocyanate compound of (3) and (4) may also be those that are distributed and sold in the market.

The compounds represented by the above formulas (1) to (4) exhibit excellent repelling activity against small mites (see Tables 6 and 7), respectively, and compounds represented by formulas (1) to (4) A mixture of two or more of them also exhibits excellent repelling activity against small mites (see Table 8).

In the present invention, the compounds may be included in a concentration of 0.1 to 10.0% with respect to the repellent composition. A preferred concentration is 0.5 to 10% concentration.

If the concentration of Saturella Montanya essential oil and Amorasia rusticana essential oil compound is less than 0.1% of the repellent composition, the repelling effect may be low, and when it exceeds 10.0% of the total weight, the content of other general additives is relatively It may be difficult to achieve the expected repelling effect in terms of the mixing ratio.

In the present invention, the repellent composition may be used in the form of one or more formulations selected from the group consisting of aerosols, sprays, liquids, emulsions, coatings, creams, capsules, ointments, and granules, preferably an aerosol It can be used in the form of formulations of agents, emulsions, sprays or creams.

The repellent comprising the active ingredient compound of Saturella montanya essential oil, Amorasia rusticana essential oil, and derivative compounds thereof according to the present invention is directly sprayed on an area where the occurrence of small Sophia mites is high, or a conventional method of treating repellents It can be sprayed directly on people and livestock or used in household products including clothing.

The repellent may be prepared and applied in various forms, and may be dissolved and dispersed in one or more solvents for spraying or application, or by using conventional surfactants as an extender such as a liquid solvent, pressurized liquefied gas, lotion and/or thickener. It is preferably used in the form of a spray, an aerosol, an emulsion, or a cream in combination with an agent.

In order to prepare such various types of preparations, the active ingredient compounds of Saturella montanya essential oil and Amorasia rusticana essential oil according to the present invention, and derivative compounds thereof, solvents and additives commonly used in the field of repellents can be used. have. In addition, the content of the active ingredient in the repellent is used in an amount effective to exhibit a repelling effect against mites, and the specific content at this time may vary depending on the form and application method of the preparation.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited by the examples.

Example 1. Extraction of essential oil from plants

In the present invention, Saturella montana essential oil used commercially sold and distributed essential oil, and Amorasia rusticana was obtained by extracting the root part by continuous steam distillation extraction method.

After washing the roots of Amorasia rusticana in a plant state with distilled water, it was ground and dried for more than 48 hours in a well-ventilated place. 1000 ml of distilled water was injected into 100 g of the pulverized sample and extracted for 3 to 6 hours at 100 to 150 ° C using a steam distillation extraction device (SDE). At this time, the extraction solvent was hexane 300, a non-polar organic solvent ml was used. The extracted hexane extract was dried over anhydrous magnesium sulfate, concentrated under reduced pressure, sealed and refrigerated.

Example 2. Repellent activity assay for small Sophia tick

Example 2-1. Small Sophie mite nymphs used in the repelling activity assay

The larvae, nymphs, and adults were used for the nymph stage showing the highest density among larvae, nymphs, and adults, and wild individuals were collected and used from the grasses near the Jeju ranch. The collected ticks were stored at a temperature of 26°C and a relative humidity of 70±5 before use.

Example 2-2. Repellent activity assay method using climbing test

A climbing test was used to test the repelling activity of the Amorasia rusticana essential oil extracted in Example 1 and the commercially available Saturella montana essential oil against small Sophie mites (see FIG. 1).

The climbing test is a method that utilizes the behavioral response of a tick that climbs vertically, and the activity that suppresses the climbing behavior can be used as an indicator of the tick's repelling effect.

After designating the upper part of the filter paper measuring 8.0 cm in length × 1.0 cm in width as the sample processing zone (height 1.0 cm) and the center part as the tick processing zone (1.0 cm in height), 20 μl of diluted samples in various concentrations were uniformly applied to the A zone. processed. The treated sample was volatilized at room temperature for 30 and 60 minutes, respectively, and then mounted on a cradle, and 10 nymphs were inoculated into the B zone. As a negative control, ethanol, which is a dilution solvent of the sample, was used.

After recording the number of mites that traversed area A where each sample was treated, the repelling rate (%) was expressed by the following equation. where T is the number of ticks that do not avoid in the sample treatment area (mite crossing the A area), and C is the number of ticks that do not avoid in the negative control group.

Evasion rate (%) = (C-T)/C × 100

표준오차로 나타내었다. The evasion rate was calculated using the SPSS statistical program (version 12.0), and the average

It is expressed as standard error.

Example 2-3. Evaluation of repelling activity of Saturella montana essential oil and Amorasia rusticana essential oil

As a result of assaying the repelling activity of Saturella montana essential oil and Amorasia rusticana essential oil using the climbing test, as shown in Table 1 below, both essential oils had excellent repelling activity of 100% up to 60 minutes at 5.0% and 3.0% concentrations. showed Table 1 below shows the results of repelling activity against small mites nymphs of Saturella montana essential oil and Amorasia rusticana essential oil using a climbing test.

sample density(%)

표준오차, %)Evasion rate (average

standard error, %) Elapsed time (min) after sample application 30 60 Saturella Montana Essential Oil 5.0 100.0±0.0 100.0±0.0 3.0 100.0±0.0 100.0±0.0 1.0 86.7±3.3 63.3±3.3 Amorasia Rusticana Essential Oil 5.0 100.0±0.0 100.0±0.0 3.0 100.0±0.0 100.0±0.0 1.0 76.7±3.3 56.7±3.3

Example 3. Component analysis and active ingredient search of Saturella montana essential oil and Amorasia rusticana essential oil

Example 3-1. Analysis of components of Saturella Montanya essential oil and Amorasia Rusticana essential oil

The components of Saturella montanya essential oil and Amorasia rusticana essential oil were analyzed using gas chromatography-mass spectrometry (Hewlett-Packard HP 6890, H5973 IV series).

HP Innowax capillary column and DB5 column (0.25 mm i.d. × 0.25 μm thickness × 2,990 cm) were used for the column, and helium was used as the carrier gas, and the temperature was 51-201°C (2°C). /min).

The temperature of the injection port (injector) was set to 211 ℃, ionization (ionization potential) was carried out at 70 eV.

The results of component analysis of Saturella montanya essential oil and Amorasia rusticana essential oil are shown in Tables 2 and 3 below, respectively.

Table 2 below shows the chemical composition of Saturella Montanya essential oil.

Retention Time (min) Library Search Composition ratio (%) 4.935 Alpha-Pinene 1.37 5.222 campen 0.13 5.735 beta-pinene 0.27 5.910 beta-myrcene 1.06 6.445 Alpha-terpinene 0.65 6.590 Osso-shimen 11.59 6.675 limonene 0.56 6.735 eucalyptol 0.48 7.230 Gamma-terpinene 1.37 7.930 linol 1.62 9.215 Bornell 0.14 9.600 Alpha-terpineol 0.32 10.585 Para-Man's-6-en-2-one 0.15 11.170 thymol 0.53 11.370 5-isopropyl-2-methylphenol 76.40 13.265 Beta-caryophyllene 2.45 13.740 Alpha-caryophyllene 0.30 15.500 Caryophyllene Oxide 0.41 26.810 13-dococenamide 0.20

As shown in Table 2 above, as a result of analyzing the compounds contained in Saturella montana essential oil, 5-isopropyl-2-methylphenol (76.40%), ortho-cymene (11.59%), beta-caryophyllene (2.45) %), linanol (1.62%), alpha-pinene (1.37%) and gamma-terpinene (1.37%) were identified.

Table 3 below shows the chemical composition of Amorasia rusticana essential oil.

Retention Time (min) Library Search Composition ratio (%) 4.841 Butyl isothiocyanate 1.43 6.035 Deccan 20.54 6.218 1,1-dimethoxycyclohexane 3.77 9.631 dodecane 2.13 13.805 Phenethylisothiocyanate 65.86 24.956 1,2-Benzenedicarboxyl acid 6.28

As shown in Table 3, as a result of analyzing the compounds contained in Amorasia rusticana essential oil, phenethylisothiocyanate (65.86%), decane (20.54%), 1,2-benzenedicabyl acid (6.28%) ), 1,1-dimethoxycyclohexane (3.77%), dodecane (2.13%) and butylisothiocyanate (1.43%) were identified.

Example 3-2. Evaluation of repelling activity of Saturella Montanya essential oil ingredients

The repelling activity of the components of Saturella montana essential oil analyzed in Example 3-1 against the small Sophie mite was assayed using a climbing test. The results are shown in Table 4 below.

In order to evaluate the repelling activity of the components of the Saturella Montanya essential oil, 5-isopropyl-2-methylphenol, ortho-cymene, beta-caryophyllene, linanol, beta-myrcene, alpha-pinene and gamma -Terpinene was purchased from Sigma-Aldrich and used. Table 4 below shows the repelling activity results of the components of Saturella Montanya essential oil against the small mites nymph using a climbing test.

sample density(%)

표준오차, %)Evasion rate (average

standard error, %) Elapsed time (min) after sample processing 30 60 5-isopropyl-2-methylphenol 1.0 100.0±0.0 83.3±3.3 Osso-shimen 1.0 6.7±3.3 0.0±0.0 Beta-caryophyllene 1.0 26.7±3.3 10.0±10.0 linol 1.0 0.0±0.0 0.0±0.0 beta-myrcene 1.0 10.0±10.0 0.0±0.0 Alpha-Pinene 1.0 0.0±0.0 0.0±0.0 Gamma-terpinene 1.0 0.0±0.0 0.0±0.0

As shown in Table 4, 5-isopropyl-2-methylphenol at a concentration of 1.0% exhibited excellent repelling activity of 83.3% up to 60 minutes. On the other hand, ortho-cymene, beta-caryophyllene, linanol, beta-myrcene, alpha-pinene and gamma-terpinene did not exhibit low or no repelling activity at a concentration of 1.0%. Therefore, it was confirmed that the active ingredient showing repelling activity of Saturella montana essential oil was 5-isopropyl-2-methylphenol.

Example 3-3. Evaluation of repelling activity of Amorasia rusticana essential oil ingredients

The repelling activity of the components of the Amorasia rusticana essential oil analyzed in Example 3-1 against the small Sophia mite was assayed using a climbing test.

In order to evaluate the repelling activity of Amorasia rusticana essential oil components, butyl isothiocyanate, decane, dodecane and phenethyl isothiocyanate were purchased from Sigma-Aldrich and used.

Table 5 below shows the repellent activity results of the components of Amorasia rusticana essential oil against the nymph of small mites using the climbing test.

sample density(%)

표준오차, %)Evasion rate (average

standard error, %) Elapsed time (min) after sample processing 30 60 Butyl isothiocyanate 1.0 13.3±3.3 0.0±0.0 Deccan 1.0 0.0±0.0 0.0±0.0 dodecane 1.0 6.7±3.3 0.0±0.0 Phenethylisothiocyanate 1.0 100.0±0.0 93.3±3.3

As shown in Table 5, 5-isopropyl-2-methylphenol at a concentration of 1.0% exhibited excellent repelling activity of 93.3% up to 60 minutes. On the other hand, butyl isothiocyanate, decane and dodecane did not show low repellent activity or no repellent activity at a concentration of 1.0%. Therefore, it was confirmed that the active ingredient showing repelling activity of Amorasia rusticana essential oil was phenethyl isothiocyanate.

Example 5. Evaluation of repelling activity of active ingredients of Saturella montana essential oil and Amorasia rusticana essential oil, and derivatives thereof

Example 5-1. Comparison of repelling activity of 5-isopropyl-2-methylphenol and its derivative compounds

Small Sophie Charm of '5-isopropyl-2-methylphenol compound' and its derivative compound '5-isopropyl-2-methylaniline', an active ingredient of Saturella Montanya essential oil confirmed in Example 3-2 Repellent activity by concentration for ticks (0.5%, 1.0%, 2.0% and 3.0%) was compared using a climbing test.

Table 6 below shows the repelling activity results of 5-isopropyl-2-methylphenol and its derivative compounds against the nymph of small mites.

sample density(%)

표준오차, %)Evasion rate (average

standard error, %) Elapsed time (min) after sample processing 30 60 5-isopropyl-2-methylphenol 3.0 100.0±0.0 100.0±0.0 2.0 100.0±0.0 100.0±0.0 1.0 100.0±0.0 83.3±3.3 0.5 63.3±5.8 26.7±5.8 5-isopropyl-2-methylaniline 3.0 100.0±0.0 100.0±0.0 2.0 86.7±8.8 63.3±6.7 1.0 76.7±8.8 40.0±5.8 0.5 56.7±5.8 23.3±5.8

As shown in Table 6, 5-isopropyl-2-methylphenol exhibited 100% repelling activity at concentrations of 3.0% and 2.0% up to 60 minutes, and 83.3% at concentrations of 1.0% and 0.5% up to 60 minutes. showed repellent activity. In addition, 5-isopropyl-2-methylaniline exhibited repelling activity of 100%, 63.3%, and 40%, respectively, up to 60 minutes at concentrations of 1.0%, 2.0%, and 3.0%.

Example 5-2. Comparison of repelling activity of phenethylisothiocyanate and its derivative compounds

Concentration (0.5%, 0.5%; The repelling activity according to 1.0%, 2.0% and 3%) was compared using the climbing test.

Table 7 below shows the repelling activity results of phenethyl isothiocyanate and its derivative compounds against small mites nymphs.

sample density(%)

표준오차, %)Evasion rate (average

standard error, %) Elapsed time (min) after sample processing 30 60 Benzylisothiocyanate 3.0 100.0±0.0 100.0±0.0 2.0 100.0±0.0 86.7±6.7 1.0 96.7±3.3 73.3±3.3 0.5 63.3±5.8 46.7±5.8 Phenethylisothiocyanate 3.0 100.0±0.0 100.0±0.0 2.0 100.0±0.0 100.0±0.0 1.0 100.0±0.0 93.3±3.3 0.5 73.3±5.8 53.3±5.8

As shown in Table 7, phenethyl isothiocyanate exhibited repelling activity of 100% at concentrations of 3.0% and 2.0% up to 60 minutes, and repelling activity of 93.3% at concentrations of 1.0% and up to 60 minutes. . Benzyl isothiocyanate exhibited repelling activity of 100% at a concentration of 3.0% up to 60 minutes, and at concentrations of 2.0% and 1.0%, repelling activity of 86.7% and 73.3%, respectively, up to 60 minutes.

Example 6. Evaluation of synergistic effect on repelling activity according to 1:1 mixing of active ingredients of Saturella montana essential oil and Amorasia rusticana essential oil and a derivative compound thereof

Repellent compounds identified in Examples 5-1 and 5-2 (1) 5-isopropyl-2-methylphenol, (2) 5-isopropyl-2-methylaniline, (3) benzyl isothiocyanate And (4) the repelling activity was tested using a climbing test to confirm the synergistic effect of repelling activity according to 1:1 mixing of each compound of phenethyl isothiocyanate, and validation of the significance of the synergistic effect of repelling activity is Duncan's multiple A range test was used ( P <0.05).

The results of comparison of repelling activity according to 1:1 mixing of the active ingredients of Saturella montana essential oil and Amorasia rusticana essential oil and their derivative compounds are shown in Table 8 below.

sample density(%)

표준오차, %)Evasion rate (average

standard error, %) Elapsed time (min) after sample processing 60 5-isopropyl-2-methylphenol (1) 0.5 26.7±5.8ef 5-isopropyl-2-methylaniline (2) 0.5 23.3±5.8f Benzylisothiocyanate (3) 0.5 46.7±5.8cd Phenethylisothiocyanate (4) 0.5 53.3±5.8bc (1)+(2) 0.5 40.0±10.0d (1)+(3) 0.5 63.3±5.8ab (1)+(4) 0.5 66.7±5.8a (2)+(3) 0.5 36.7±5.8de (2)+(4) 0.5 43.3±5.8cd (3)+(4) 0.5 70.0±10.0a

As shown in Table 8, repelling activity and (1)+(2), (1)+(3), (1)+(2), (1)+(3), (1)+(4), (2)+(3), and (3)+(4) mixture (mixing 1:1 weight ratio) after 60 minutes compared the repelling activity, compound (1)+( 2), (1) + (3), (1) + (4) and (3) + (4) in the mixture was confirmed a significant increase in repelling activity than each alone treatment (see FIG. 2).

Claims (6)

5-isopropyl-2-methylphenol (1), a substance of Saturella Montanya essential oil, 5-isopropyl-2-methylaniline (2), a derivative of this substance, phenethyliso, a substance of Amorasia rusticana essential oil A repellent active composition for controlling small mites, characterized in that it contains at least one compound selected from among thiocyanate (3) and benzyl isothiocyanate (4), which is a derivative thereof, as an active ingredient.
According to claim 1,
The compound is characterized in that it contains 0.1 to 30.0% concentration of the composition, repellent active composition for controlling small mites The method of claim 1,
The active ingredient is a mixture of 5-isopropyl-2-methylphenol (1) and 5-isopropyl-2-methylaniline (2); a mixture of 5-isopropyl-2-methylphenol (1) and phenethylisothiocyanate (3); a mixture of 5-isopropyl-2-methylphenol (1) and benzylisothiocyanate (4); a mixture of 5-isopropyl-2-methylaniline (2) and phenethylisothiocyanate (3); a mixture of 5-isopropyl-2-methylaniline (2) and benzylisothiocyanate (4); Or a mixture of phenethyl isothiocyanate (3) and benzyl isothiocyanate (4); 4. The method of claim 3,
The ratio of the components of each mixture is 1:1 by weight, characterized in that the repellent active composition for controlling small Sophie mites A mite repellent prepared using the repellent active composition for controlling a small sophia mite according to any one of claims 1 to 4. 6. The method of claim 5,
The repellent is a mite repellent, characterized in that it is used in the form of one or more formulations selected from the group consisting of aerosols, emulsions, sprays and creams.

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