WO2003037087A1

WO2003037087A1 - Pesticide formulation composed of natural components

Info

Publication number: WO2003037087A1
Application number: PCT/KR2002/001897
Authority: WO
Inventors: Jeong Joo Shin; Sang Jun Kim; Soon Jong Han; Jae Min Ha; Min Hyuk Lee
Original assignee: Sk Chemicals Co., Ltd.
Priority date: 2001-10-10
Filing date: 2002-10-10
Publication date: 2003-05-08
Also published as: KR20030030349A; KR100834965B1; CN1568145A

Abstract

The present invention relates to a natural pesticide composition. More particularly, the present invention relates to a natural pesticide composition comprising a yu teng extract and ku shen extract, which may further comprise other herbal medicine extracts obtained from Meliae cortex, Meliae fructus, Azadirachta indica and the like, if deemed necessary, thus having an excellent pest control activity against various kinds of agriculturally noxious insects such as TSSM (two spotted spider mite, Tetranychus urticae), DBM (Diamond-back moth, Plutella xylostella) which have been known very hard to be placed under effective pest control. Further, the pesticide composition is shown harmless to humans and livestock animals.

Description

PESTICIDE FORMULATION COMPOSED OF NATURAL COMPONENTS

[TECHNICAL FIELD]

The present invention relates to a natural pesticide composition. More particularly, the present invention relates to a natural pesticide composition comprising a yu teng extract and a ku shen extract, which may further comprise other herbal medicine extracts obtained from Kulianpi, Chuanlianzi, Neem seed and the like, if deemed necessary, thus having an excellent pest control activity against various kinds of agriculturally noxious insects such as TSSM(two spotted spider mite, Tetranychus urticae), DBM(Diamond-back moth, Plutella xylostella) which have been known very hard to be placed under effective pest control. Further, the natural pesticide composition is shown harmless to humans and livestock animals.

[BACKGROUND ART]

Environmental pollution including soil and water pollution resulted from extensive spray of various synthetic pesticides has been raised as one of the most serious environmental problems. Therefore, researches have been mainly focused on developing natural pesticides with low toxicity or not causing pollution problems. Namely, natural pesticides that are harmless to humans and can protect agricultural products from damages by harmful insects are presently in urgent demand.

Researches using yu teng and ku shen as raw materials of a pesticide have been conducted since 1980s [Chinese Patent Nos. 1176742, 1183902, 1057159, 1194782, 97104754, 1171198]. For example, a natural pesticide containing 0.08% ku shen alkaloid was disclosed in Chinese Patent No. 1171198. Also, ku shen of plants and tobacco was described as a raw material for a pesticide composition in Chinese Patent No. 1194782, in which pharmaceutical effects were obtained via a simple process consisting of a crash extraction and mixing of extracts, which was used for the purpose of pest control in vegetables, fruits and cotten plants, particularly directed to controls against cabbage butterfly, pieris rapae larvae and nests of aphides.

However, these conventional methods were largely limited in their pest control activities, especially in controlling against TSSM and DBM with good effect, which are classified as harmful insects hard to be eliminated under conventional treatments.

Information on yu teng and ku shen has been well provided via the encyclopedia of herbal medicines, such as A colorful illustrated guide of Chinese herbal medicines, Pharmacopoeia of People's Republic of China, the encyclopedia of toxic Chinese herbal medicines and Poisonous medical botany. The above herbal medicines have been used for treating symptoms of dropsy, diuresis, antiarrhythmic and the like resulted from various strokes and also have been once applied to agriculture due to their known pesticidal activity. From the pathological point of view, those herbal medicines are known to stimulate central nervous system and also cause paralytic poisoning.

The present invention contains yu teng and ku shen extracts, which have been used as agricultural pesticides according to folk remedies, and thus have a wide insecticidal spectrum as well as excellent pest control activities with selectivity. Further, extracts obtained from Kulianpi, Chuanlianzi and Neem seed can be further added to yu teng and ku shen extracts to maximize the desired effects. Therefore, an object of this invention is to provide a novel composition of a natural pesticide which has excellent pest control activity against noxious insects which are hard to be controlled under treatments of conventional pesticides.

[DISCLOSURE OF INVENTION]

The present invention is characterized by a natural pesticide composition comprising an effective amount of yu teng extract and an effective amount of ku shen extract, respectively.

Further, the natural pesticide composition of this invention may comprise further at least one extract selected from the group consisting of Kulianpi, Chuanlianzi and Neem seed extracts along with yu teng extract and ku shen extract.

The characteristics of natural plants used as raw materials in this invention are summarized as follows. Yu teng is a leguminous plant where all parts of the plant or its root can be used as a raw material for herbal medicines and is known to be well acclimated to tropical or subtropical weather. Yu teng grows under a relatively shadowy condition and is preferred to grow on a thick-layered soil with good drainage. The main habitats of yu teng include Kwangdong, Kwangseo, Wunnam, Bokgun, Honam, Henam provinces in China and Taiwan, and yu teng produced in Kwangdong and Haenam are especially considered to have high qualities. The roots of yu teng contain extremely toxic materials and are thus able to kill or poison insects. Yu teng is able to kill hundreds different kinds of insects and with rapidity. According to the result s obtained by Department of Plant Protection in Hwanam Institute of Agriculture(China), yu teng is easily decomposed by exposure to light and easily oxidized in the air. Therefore, there are no materials left as a residue after use and also it will not cause any environmental pollution. Further, it is shown very toxic only to insects and fish, but not to mammals.

Ku shen is a dried root of Sophora flavescens and produced throughout the mainland of China. In general, products from Sanseo, Hobuk, Hanam and Habuk provinces in China are considered excellent. It is acclimated to warm weather and thick layered, fertile, well-drained sandy soil or clay soil. It is preferred to collect ku shen in spring and autumn, more preferably in autumn. Roots exhumed to remove the upper part of root and fibrous root are rinsed with water to get rid of mud and sand, and then dried in the sun. Ku shen sliced and dried in the sun is called 'ku shen pyun'.

Chuanlianzi is a dried fruit of Fructus toosendan and largely distributed through the area of Suchuan, Hobei, Guiju, Hanam, Anhui, Jiangsu provinced in China and the like. The Kulianpi is of a spherical shape with a size of 2 - 3.2 cm in diameter and the surface takes on a color of crimson yellow or brown yellow. Kulianpi is a little glossy, and usually dented or shrunk on the surface. Its inside is partitioned into 6 - 8 parts and there is a brown oval seed in each part. It has a unique smell tastes sour.

Kulianpi is a bark of dried tree of Melia toosendan sieb. Its main habitats are Suchuan, Hubei, Anhui, Jiangsu, Hanam, Guiju, Shandong and Yuannan provinces in China. Kulianpi contains toosendarin which tastes bitter. It has a shape of an irregular flat plate and its tube and width are irregular. It has a pattern of crossing vertical lines and yellow brown bark holes. When the rough bark of Kulianpi is removed, there appears white or yellow white inside.

Neem seed of the Azadirachta indica, is thick of leaves and branches and belongs to a tropical forest tree whose trunk is high and green. It grows rapidly and is highly tolerant of extreme conditions such as drought, high temperature and sterile conditions. In general, the higher the trunk grows, the deeper the root digs into the ground. The original habitats of Neem seed are India, Myanmar and Pakistan. However, they have been implanted and are now cultivated in Thai, Iran, Dominica and China. Neem seed contains an active ingredient called azadirachtin known to have various pharmaceutical effects.

Natural plants described above are utilized in the present invention by means of some samples prepared according to standard methods. First, external dirts and pollutants are removed from the natural plants, and then sliced or crushed for easier extraction. Unlike conventional methods used for extracting a single or combined herbal medicines, the present invention conducted the extraction using solvents classified into two different groups of polar and non-polar solvents, wherein all the herbal medicines used in the present invention were determined to use one of the solvents grouped based on the characteristics of each herbal medicine.

Yu teng extract was prepared by using conventional non-polar organic solvents such as benzene, toluene, xylene, cyclohexane, n-hexane, n-pentane, n- heptane, dichloromethane, chloroform, tetrachloromethane and the like, and they can be used alone or as a mixture of the solvents. However, for maximizing the extraction efficiency of pesticidal ingredients obtained from yu teng, solvents should be selected carefully. In order to prevent decomposition of active ingredients, the extraction was performed at a temperature below 80 ^°C . Yu teng was extracted three times for 7 hr, respectively, the combined extracts were filtrated and concentrated. This conventional extraction method can be used in the present invention.

In contrast, ku shen extract was prepared by using conventional polar organic solvents such as methanol, ethanol, isopropanol, n-propanol, benzyl alcohol, propylcellulose, γ-butylolactone, N-methyl-2-pyrolidone, methyl naphthalene, methyl ether fatty acid and the like and they can be used alone or as a mixture of the solvents. For maximizing the extraction efficiency of pesticidal ingredients obtained from ku shen, solvents should be selected carefully. The extraction was performed at a temperature below 80 ^°C to avoid decomposition of active ingredients. The extraction was performed 3 times for 5 hr, respectively, followed by filtration of the combined extracts and concentration in this order. This conventional method can be used in the present invention.

In the present invention, Kulianpi, Chuanlianzi and Neem seed were extracted using the methods most appropriate for each. Kulianpi and Chuanlianzi were extracted by using the same method for yu teng, whereas Neem seed was extracted by using the same method for ku shen.

The natural pesticide composition of the present invention comprises 20 - 80 wt% of yu teng extract(with reference to 3.0 wt% of rotenone content, as a marker substance) and 20 - 80 wt% of ku shen extract(with reference to 0.2 wt% of matrine alkaloid content, as a marker substance) as essential ingredients. The natural pesticide composition of the present invention may further comprise Kulianpi, Chuanlianzi and Neem seed (with reference to 7.0 wt% of the extract content) in the range of 0 - 30 wt%. If the content of yu teng extract or ku shen extract is below 20 wt% or more than 80 wt%, it is hard to effectively control both TSSM(two spotted spider mite, Tetranychus urticae) and DBM(Diamond-back moth, Plutella xylostella). A pesticidal composition containing only yu teng and ku shen extracts without extracts from Kulianpi, Chuanlianzi and Neem seed can exhibit excellent pesticide control. However, the pesticidal composition is preferred to further comprise Kulianpi, Chuanlianzi and Neem seed as active ingredients for exhibition of better pesticidal activities, particularly, for acquiring excellent pest control activities against noxious insects such as TSSM and DBM. If Kulianpi, Chuanlianzi and Neem seed are contained more than 30 wt%, it results in limiting the original pest control property conferred by yu teng and ku shen.

In the case of herbal medicines prepared by using extracts obtained from natural plants, the contents of active ingredients vary greatly depending on the place of production, time of collection, period of storage and storage conditions, thus it is preferred to choose an appropriate marker substance for each extract and determine the range of the content of each ingredient for a pesticide composition with reference to the content of a predetermined marker substance. In the present invention, as a marker substance, rotenone was selected for yu teng, and matrine for ku shen, respectively. In the case of herbs such as Melia toosendan sieb, Fructus toosendan and Neem seed, which may be added additionally, were selected based on the wt% of the content of each extract.

Therefore, the range of the content of each ingredient for a pesticide composition can be determined in accordance with the content rate of a marker substance and extract. Accordingly, the natural pesticide composition according to the present invention comprises 0.6 - 2.4 g of rotenone, 0.04 - 0.16 g of matrine and 0 - 2.1 g of additional extract content of Kulianpi, Chuanlianzi or Neem seed with reference to 100 g of the natural pesticide composition. In the natural pesticide composition of the present invention, the reason for restricting the content of each active ingredient is the same as in the above-described natural plant extracts.

As explained above, the natural pesticide composition according to the present invention comprises yu teng extract(rotenone) and ku shen extract(matrine), thus having an excellent pesticidal effect, and may further comprise Kulianpi, Chuanlianzi and Neem seed, thus imparting an increased pesticidal effect, particularly against TSSM(two spotted spider mite, Tetranychus urticae) and DBM(Diamond-back moth, Plutella xylostella) which are known very hard to be placed under effective pest control under conventional methods. As explained above, the present invention provides crude plant extracts as shown in the following preparation examples, and natural pesticide compositions by the following examples. Pesticidal activities were examined by using experimental methods for the natural pesticide compositions prepared by each example. The present invention will be further illustrated by the following preparation examples and examples, however, they should not be construed as limiting the scope of the present invention.

[EXAMPLES] Preparation Example 1 : Preparation of yu teng extract

Yu teng was washed with water and dried, and then it was sliced and crushed. 440 g of the crushed yu teng was added with 1,365 ml of toluene and then incubated at 50 ^°C for 7 hr, and an extraction was performed at 70 ^°C for 10 hr while refluxing. Then, the extract was collected, and 1,090 ml of toluene was added to the residue, and another extraction was performed at 70 ^°C for 6 hr and the extract was collected. Again, 1,000 ml of toluene was added to the residue and the third extraction was performed at 70 ^°C for 2 hr and the extract was collected. All the above extracts were combined and concentrated to 210 ml.

Preparation Example 2 : Preparation of ku shen extract

58 g of ku shen sliced to 2.0 - 4.0 cm was mixed and incubated in 410 ml of 90 v/v% ethanol at 50 ^°C for 14 hr, and an extraction was performed at 70 ^°C for 3 hr while refluxing. Then, the extract was collected, and 275 ml of 90 v/v% ethanol was added to the residue, and another extraction was performed at 70 ^°C for 6 hr and the extract was collected. The third extraction was performed for the residue at 70 ^°C for 2 hr and the extract was collected. All the above extracts were combined and concentrated to 210 ml.

Preparation Example 3 : Preparation of Kulianpi extract

Kulianpi was washed with water and dried, and then it was sliced and crushed. 440 g of the crushed Kulianpi was added with 1,365 ml of toluene and then incubated at 50 ^°C for 7 hr, and an extraction was performed at 70 ^°C for 10 lir while refluxing. Then, the extract was collected, and 1,090 ml of toluene was added to the residue and another extraction was performed at 70 ^°C for 6 hr and the extract was collected. Again, 1,000 ml of toluene was added to the residue and the third extraction was performed at 70 ^°C for 2 hr and the extract was collected. All the above extracts were combined and concentrated to 210 ml.

Preparation Example : Preparation of Chuanlianzi extract

Chuanlianzi was washed with water and dried, and then it was sliced and crushed. 440 g of the crushed Chuanlianzi was added with 1,365 ml of toluene and then incubated at 50 ^°C for 7 hr, and an extraction was performed at 70 ^°C for 10 hr while refluxing. Then, the extract was collected, and 1,090 ml of toluene was added to the residue and another extraction was performed at 70 ^°C for 6 hr and the extract was collected. Again, 1,000 ml of toluene was added to the residue and the third extraction was performed at 70 ^°C for 2 hr and the extract was collected. All the above extracts were combined and concentrated to 210 ml.

Preparation Example 5 : Preparation of Neem seed extract 58 g of Neem seed sliced to 2.0 - 4.0 cm was evenly mixed and incubated in 410 ml of 90 v/v% ethanol at 50 ^°C for 14 hr, and then an extraction was performed at 70 °C for 3 hr while refluxing. Then, the extract was collected, and 275 ml of 90 v/v% ethanol was added to the residue, and another extraction was performed at 70 °C for 6 hr and the extract was collected. The third extraction was performed for the residue at 70 ^°C for 2 hr and the extract was collected. All the above extracts were combined and concentrated to 210 ml.

Preparation Example 6 : Preparation of yu teng and Kulianpi extracts The mixture of 440g of well-dried yu teng and 15 g of well-dried

Kulianpi was added with 1,365 ml of toluene, and then incubated at 50 ^°C for 7 hr, followed by extraction at 70 °C for 10 hr while refluxing. Then, the extract was collected, and 1,090 ml of toluene was added to the residue and another extraction was performed at 70 ^°C for 6 hr and the extract was collected. Again, 1,000 ml of toluene was added to the residue and the third extraction was performed at 70 °C for 2 hr and the extract was collected. All the above extracts were combined and concentrated to 210 ml. Preparation Example 7 : Preparation of yu teng and Chuanlianzi extracts

The mixture of 440g of well-dried yu teng and 58 g of well-dried Chuanlianzi was added with 1,365 ml of toluene, and then incubated at 50 ^°C for 7 hr, followed by an extraction performed at 70 ^°C for 10 hr while refluxing. Then, the extract was collected, and 1,090 ml of toluene was added to the residue and another extraction was performed at 70 ^°C for 6 hr and the extract was collected. Again, 1,000 ml of toluene was added to the residue and the third extraction was performed at 70 ^°C for 2 hr and the extract was collected. All the above extracts were combined and concentrated to 210 ml.

Preparation Example 8 : Preparation of yu teng, Chuanlianzi and Kulianpi extracts

The mixture of 440g of well-dried yu teng, 58 g of well-dried Chuanlianzi and 15 g of well-dried Kulianpi was added with 1,365 ml of toluene, and then incubated at 50 ^°C for 7 hr, followed by an extraction performed at 70 ^°C for 10 hr while refluxing. Then, the extract was collected, and 1,090 ml of toluene was added to the residue and another extraction was performed at 70 ^°C for 6 hr and the extract was collected. Again, 1,000 ml of toluene was added to the residue and the third extraction was performed at 70 ^°C for 2 hr and the extract was collected. All the above extracts were combined and concentrated to 210 ml.

Preparation Example 9 : Preparation of ku shen and Neem seed extracts

58 g of ku shen sliced to 2.0 - 4.0 cm is evenly mixed with 30 g of Neem seed which is crushed so that kernel of the seed is broken, and incubated in 410 ml of 90 v/v% ethanol at 50 ^°C for 14 hr, and then an extraction was performed at 70 °C for 3 hr while refluxing. Then, the extract was collected, and 275 ml of 90 v/v% ethanol was added to the residue, and another extraction was performed at 70 °C for 6 hr and the extract was collected. The third extraction was performed for the residue at 70 ^°C for 2 hr and the extract was collected. All the above extracts were combined and concentrated to 210 ml.

Reference Example : Selection of a marker substance Solvent gradient pump (Waters 600 Controller, USA) was used for HPLC analysis of rotenone, a marker substance for yu teng extract. The solvent used was 0.1 N aqueous solution of phosphoric acid for acetonitrile, where gradient was allowed ranging from 90:10 to 0:100 for 90 min. Phenomenex Cis was used as column, and UV detector was set at the wavelength of 295 ran. Samples were added as much as 10 μJL each time using an auto sampler. The result was analyzed by using Autochro software (Younglin Co., Korea).

In a separating funnel, 1.6203 g of alkaloid matrine, a marker substance for ku shen, was mixed with 2 ml of concentrated aqueous ammonia and 10 ml of chloroform. Extractions were repeatedly performed three times, followed by evaporation in water bath, respectively. The details of evaporation are as follows: Evaporation was repeated for the residue by using 3 ml of anhydrous ethanol. 10 ml of ethylether was added so to dissolve the extract, and then 10 ml of 0.01 M sulfuric acid was added to evaporate ether. After cooling, the extract was added with 15 ml of distilled water and 2 - 3 drops of methyl orange indicator and titrated by using 0.0176 M sodium hydroxide. For a blank test, 10 ml of 0.01 M sulfuric acid, 15 ml of distilled water and 2 - 3 drops of methyl orange were added.

Marker substances selected by the above-mentioned methods are as follows.

Matrine

Example : Preparation of natural pesticide composition

Natural plant extracts prepared from the above preparing examples were mixed at the rates shown in Table 1. Meanwhile, polyoxyethylene sorbitan monoolate("emulsifier 1", Koremul SFT-85, Hannong Hwasung, Korea) and polyoxyethylene octylphenylether("emulsifier 2", Koremul OP-10, Hannong Hwasung, Korea) were used as much as 6 wt% and 4 wt%, respectively, to the sum of the natural extracts for the homogeneous emulsification of the composition. Ethanol for industrial use(Jin Chemical Pharmaceutical Co., Ltd.) was used as a solvent to adjust the final volume of the composition to 100%. [Table la]

[Table lb]

Experimental Example

Experiments were performed as shown below in order to compare the activities of the natural pesticide compositions prepared according to the composition in Tables la and lb.

. , , _{rr m},. number of dead insects , . _ pesticidal effect(%) = x lOO number of inoculated insects

Experimental Example 1 : Pharmaceutical effect against TSSM

Twenty imagos of TSSM(two spotted spider mite, Tetranychus urticae) placed on disc of kidney bean leaves of 3 cm in diameter. Sprayed with a predeterminated amount of a composition, followed by drying in the dark in hood. 24 lir and 48 hr after the spray, respectively, the pesticidal effects were examined and the result are shown in the following Table 2.

Experimental Example 2 : Pharmaceutical effect against DBM Cabbage leaves of 5 cm in diameter were soaked in the prepared composition for 30 sec and then dried in the dark in hood, and inoculated 20 three instar larvae of DBM(diamond-back moth, plutella xylostella). 24 hr and 48 hr after the inoculation, respectively, the pesticidal effects were examined and the results are shown in the following Table 3. [Table 2] Pharmaceutical effect against TSSM [pesticidal effect(%)J

[Table 3] Pharmaceutical effect against DBM [pesticidal effect(%)]

[INDUSTRIAL APPLICABILITY]

As explained above, the present invention provides a natural pesticide composition consisting of herbal medicine extracts, which are shown harmless to both humans and livestock animals, as well as having excellent pest control activities against agriculturally noxious insect, such as TSSM and DBM known hard to be placed under effective pest control. Therefore, the natural pesticide composition of the present invention is expected to serve as a good pesticide for next generation.

Claims

[CLAIMS]What is claimed is:

1. A natural pesticide composition comprising an effective amount of yu teng extract and an effective amount of ku shen extract.

2. The natural pesticide composition according to claim 1, wherein said yu teng extract and said ku shen extract are contained 20 - 80 wt%, respectively.

3. The natural pesticide composition according to claim 1 or claim 2, wherein rotenone is contained 0.6 - 2.4 g and matrine is contained 0.04 - 0.16 g as marker substances with reference to 100 g of said natural pesticide composition.

4. The natural pesticide composition according to claim 1, wherein said composition further comprises at least one extract selected from the group consisting of Kulianpi, Chuanlianzi and Neem seed extracts.

5. The natural pesticide composition according to claim 4, wherein said Kulianpi, Chuanlianzi and Neem seed extracts are contained 0 - 30 wt% of said natural pesticide composition.

6. The natural pesticide composition according to claim 4 or claim 5, wherein rotenone is contained 0.6 - 2.4 g and matrine is contained 0.04 - 0.16 g as marker substances with reference to 100 g of said natural pesticide composition, and further said Kulianpi, Chuanlianzi and Neem seed extracts are contained 0 - 2.1 g, respectively.

7. A natural pesticide composition which is effective against both two spotted spider mite(Tetranychus urticae) and Diamond-back moth(Plutella xylostella).