Trifluoroethyl phenyl sulfide compounds, pesticidal compositions and use thereof

Abstract

The application provides a trifluoroethyl phenyl sulfide compound, a pesticide composition and application thereof, and belongs to the field of pesticides.The chemical name of the trifluoroethyl phenyl sulfide compound is (4-(4-chloro-2-fluoro-5-((2,2,2-trifluoroethyl) methyl sulfan) phenoxy) butyl) methyl sulfan.The pesticide composition comprises the trifluoroethyl phenyl sulfide compound or a salt thereof, and the pesticide composition can be used as an insecticide or a repellent for preventing and treating crop pests and / or harmful mites.The pesticide composition further comprises a biological safety agent with an inner core of a GST enzyme-NAC load, a middle layer of a nanocapsule shell and an outer layer of a composite coating.Compared with existing insecticides / miticides, the activity of the trifluoroethyl phenyl sulfide compound disclosed in the application is significantly improved, and the virulence intensity of target pest control is enhanced.The biological safety agent is added in the pesticide composition, and the toxicity of the pesticide to non-target organisms can be extremely significantly and dose-dependently reduced, and the biological safety is improved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pesticides, and particularly relates to a trifluoroethyl phenyl sulfide compound, a pesticide composition and application thereof. BACKGROUND

[0002] The compound containing a trifluoroethyl sulfide (sulfoxide) fragment as shown in the following formula (I) has good killing activity on red spider mites, aphids, brown planthoppers, two-spotted spider mites, citrus red spider mites and red spider mites and other pests or harmful mites; (I), n=0 or 1, R1 and R2 are halogen or methyl; the structures of such compounds include diazoles, triazoles, biphenyls, phenylamidines, phenyl ethers, pyrimidinones and the like. The compounds currently widely used include flumetover, pyridaben, flufenerim, and so on. It is found through biological activity tests that the above-mentioned compounds have excellent insecticidal and miticidal activity, but have the following shortcomings: (1) a high dosage is required; (2) it is difficult to achieve insect and mite control at the same time; (3) it is mainly used for killing insects and mites, and has poor prevention effect on insects and mites. In addition, since the action mechanism of the compounds is single, and the effect depends on the trifluoroethyl sulfide (sulfoxide) fragment, insects and mites can produce resistance and tolerance by enhancing the metabolic detoxification capacity under the selection pressure. Therefore, the resistance problem of many existing insect and mite control agents is increasingly serious, and even mutual resistance of pests and mites is produced. SUMMARY

[0003] In view of this, the present application aims to provide a trifluoroethyl phenyl sulfide compound, a pesticide composition and application thereof, and aims to solve at least one technical problem in the background art.

[0004] The present application is implemented in the following manner:

[0005] The present application provides a trifluoroethyl phenyl sulfide compound, which has the chemical name of (4-(4-chloro-2-fluoro-5-((2,2,2-trifluoroethyl) methyl sulfanyl) phenoxy) butyl) methyl sulfanyl, and has the following structure:

[0006] .

[0007] The present application provides the application of the above-mentioned trifluoroethyl phenyl sulfide compound, and the above-mentioned trifluoroethyl phenyl sulfide compound is used for preparing a pesticide composition.

[0008] Further, the pesticide composition is used as an insecticide for controlling crop pests and / or harmful mites; or, the pesticide composition is used as a repellent for repelling crop pests and / or harmful mites.

[0009] Further, the crop pests are at least one of lepidopteran pests, dipteran pests and lepidopteran pests.

[0010] The third aspect of the present application provides a pesticide composition comprising the above-mentioned trifluoroethyl phenyl sulfide compound or salt thereof and an agriculturally acceptable formulation carrier or formulation adjuvant; the active ingredient of the pesticide composition comprises the trifluoroethyl phenyl sulfide compound or salt thereof.

[0011] Further, the pesticide composition is an insecticide for controlling crop pests and / or mites; or the pesticide composition is a repellent for repelling crop pests and / or mites.

[0012] Further, the pesticide composition further comprises a biological safety agent in the form of a microcapsule with a core-shell structure, the core of which is a GST enzyme-NAC load, the intermediate layer is a nanocapsule shell, and the outer layer is a composite coating; the GST enzyme is glutathione S-transferase, and the NAC is N-acetylcysteine.

[0013] Further, the biological safety agent: trifluoroethyl phenyl sulfide compound or salt thereof = 0.5-1.5:1 by mass ratio.

[0014] Further, the preparation method of the biological safety agent comprises the following steps:

[0015] S1, a GST enzyme solution and an NAC solution are respectively prepared with PBS buffer as the solvent; after pectin is dissolved in hot water and cooled to room temperature, the NAC solution is mixed with the pectin to prepare an NAC / pectin solution;

[0016] S2, a chitosan-acetic acid solution is mixed with the NAC / pectin solution by ion crosslinking method to form a hydrogel microsphere wrapping NAC;

[0017] S3, the GST enzyme solution is loaded into the hydrogel microsphere prepared in S2 by diffusion method to obtain a hydrogel microsphere loaded with GST enzyme-NAC;

[0018] S4, in an ethanol and ammonia system, a hydrolysis and condensation reaction is carried out on the surface of the hydrogel microsphere prepared in S3 using tetraethyl orthosilicate to form a silica shell, and a microcapsule is obtained;

[0019] S5, in a Tris-HCl buffer, a polydopamine layer is formed on the surface of the silica shell by self-polymerization of dopamine; and chitinous oligosaccharides are connected to the polydopamine layer by coupling reaction;

[0020] S6, centrifugal washing is performed to remove unreacted systems, and a dry nanometer powder is obtained by vacuum freezing, which is the biological safety agent.

[0021] Further, the GST enzyme solution further comprises trehalose with a final concentration of 1%-5%.

[0022] Compared with the prior art, the present application comprises the following beneficial effects:

[0023] 1. The present application discloses a trifluoroethyl phenyl sulfide compound, which has significantly improved activity compared to existing insecticides and miticides, and enhances the virulence intensity of pest target control.

[0024] 2. The trifluoroethyl phenyl sulfide compound of the present application still has strong biological activity at low concentration, effectively controls and repels Lepidoptera pests, Diptera pests, Lepidoptera pests, and mites, thereby effectively reducing the amount of pesticide used.

[0025] 3. In the pesticide composition containing the trifluoroethyl phenyl sulfide compound, the addition of the biological safety agent with the inner core of GST enzyme-NAC carrier can significantly and dose-dependently reduce the toxicity of pesticide FW-10 to soil organisms, natural enemy insects, pollinating insects, and aquatic organisms, and improve biological safety.

[0026] 4. After adding the biological safety agent to the pesticide composition containing the trifluoroethyl phenyl sulfide compound, the insecticidal activity, miticidal activity, and repellent activity of the pesticide itself are not significantly affected. DETAILED DESCRIPTION

[0027] In order to make the purpose, technical scheme and advantages of the present application clearer, the present application will be further described in detail below with examples. It should be understood that the specific examples described herein are only used to explain the present application and do not limit the present application.

[0028] Example 1

[0029] This example is a trifluoroethyl phenyl sulfide compound, the chemical name of which is: (4-(4-chloro-2-fluoro-5-((2,2,2-trifluoroethyl) methyl sulfan) phenoxy) butyl) methyl sulfan, which is hereinafter referred to as FW-10; FW-10 has the following structure:

[0030] ;

[0031] The synthesis route is as follows:

[0032] .

[0033] The preparation method of the trifluoroethyl phenyl sulfide compound comprises steps 1 to 7.

[0034] Step 1, preparation of N,N-dimethylformamido-4-chloro-2-fluorobenzene;

[0035] In a 3L four-necked flask equipped with a stirrer, reflux condenser, thermometer and dropping funnel, 2-fluoro-4-chloro-phenol 293g, 4-dimethylaminopyridine 12.2g, dimethylcarbamoyl chloride 258g and dichloromethane 1.43L were added and stirred. After the mixture was dissolved, triethylamine 263g was dropped into the reaction system within 1h, and then stirred at room temperature for 5h. After confirming the end of the reaction, water 400mL and 35% hydrochloric acid 62.5g were added to the reaction system and stirred at room temperature for 30min. Subsequently, the organic layer was separated from the aqueous layer, water 450mL and 5% sodium bicarbonate aqueous solution 381g were added to the organic layer, and stirred at room temperature for 30min. The organic layer was separated from the aqueous layer again, and the obtained organic layer was distilled off under reduced pressure, and the obtained residue was recrystallized from isopropanol 400mL and water 600mL. After filtration and drying, the product was obtained as a white crystal in a yield of 93%. The product was subjected to an analysis.

[0036] 1 H-NMR (300MHz, CDC13) δ (ppm) : 7.19-7.08 (m, 3H), 3.11 (s, 3H), 3.02 (s, 3H); melting point 47.6°C.

[0037] Step 2, Preparation of bis (5-dimethylcarbamoyloxy-2-chloro-4-fluorophenyl) disulfide;

[0038] In a 300mL four-necked flask equipped with a stirrer, reflux condenser, thermometer and dropping funnel, dichloromethane 90mL and aluminum chloride 60.0g were added and stirred, and a mixed solution of N,N-dimethylcarbamoyl-4-chloro-2-fluorophenyl ester (0.30mol) prepared in Step 1 and dichloromethane 60mL was dropped at room temperature. Subsequently, at 40°C, disulfur dichloride 30.4g was dropped for 3h, and then stirred for 1h to obtain a reaction mixture.

[0039] In a 1L four-necked flask equipped with a stirrer, reflux condenser, thermometer and dropping funnel, water 279mL and dichloromethane 121mL were added, and the above obtained reaction mixture was dropped at room temperature, and then stirred for 30min. The organic layer was separated from the aqueous layer, and the organic layer was distilled off under reduced pressure to collect the residue. Toluene 300mL was added to the obtained residue, and then water 124mL, 35% hydrochloric acid 156g were added at 60°C and stirred for 30min, and the layers were separated. The organic phase was washed with 5% sodium bicarbonate aqueous solution 286g and 15% salt water 279g, respectively, to obtain a toluene solution of bis (5-dimethylcarbamoyloxy-2-chloro-4-fluorophenyl) disulfide as a product, which was directly used in the next step. A small amount of the solution was distilled off and used for detection.

[0040] 1H-NMR (300 MHz, CDC13) δ (ppm): 7.45 (d, J = 7.8 Hz, 2H), 7.21 (d, J = 9.3 Hz, 2H), 3.10 (s, 6H), 3.00 (s, 6H). Melting point is 140.4 °C. LC-MS (ESI): = 496.99 [M+H] + .

[0041] Step 3, preparation of N,N-dimethylcarbamoyl-4-chloro-2-fluoro-5-mercapto phenyl ester;

[0042] A toluene solution of the bis(5-dimethylcarbamoyloxy-2-chloro-4-fluorophenyl) disulfide (0.15 mol) prepared in step 2 was taken out and added to a 1 L four-necked flask equipped with a stirrer, reflux condenser, thermometer and dropping funnel, cooled to 10 °C, and then water 75 mL, sodium carbonate 31.8 g and 50% aqueous tetrabutylammonium bromide solution 1.9 g were added in turn, and then sodium formaldehyde sulfoxylate (Rongalit) 70 g was dissolved in water 300 mL and added dropwise into the reaction system in 1 h, and stirred at 10 °C for 1 h. Then 35% hydrochloric acid 46 g was added to adjust the pH of the reaction system to 6-7. After standing, the product N,N-dimethylcarbamoyl-4-chloro-2-fluoro-5-mercapto phenyl ester was obtained in toluene solution, and the yield was 70.6% by content analysis, which was directly used in the next step. A small amount of the solution was removed for detection.

[0043] 1 H-NMR (300 MHz, CDC13) δ (ppm): 7.45 (d, J = 7.8 Hz, 2H), 7.21 (d, J = 9.3 Hz, 2H), 3.10 (s, 6H), 3.00 (s, 6H). Melting point is 140.4 °C. LC-MS (ESI): = 496.99 [M+H]

[0044] Step 4, preparation of 4-chloro-2-fluoro-5-mercapto phenol;

[0045] A toluene solution of the bis(5-dimethylcarbamoyloxy-2-chloro-4-fluorophenyl) disulfide (0.15 mol) prepared in step 2 was taken out and added to a 1 L four-necked flask equipped with a stirrer, reflux condenser, thermometer and dropping funnel, cooled to 10 °C, and then water 75 mL, sodium carbonate 31.8 g and 50% aqueous tetrabutylammonium bromide solution 1.9 g were added in turn, and then sodium formaldehyde sulfoxylate (Rongalit) 70 g was dissolved in water 300 mL and added dropwise into the reaction system in 1 h, and stirred at 10 °C for 1 h. Then 35% hydrochloric acid 46 g was added to adjust the pH of the reaction system to 6-7. After standing, the product N,N-dimethylcarbamoyl-4-chloro-2-fluoro-5-mercapto phenyl ester was obtained in toluene solution, and the yield was 70.6% by content analysis, which was directly used in the next step. A small amount of the solution was removed for detection.

[0046] Into a 250 mL four-necked flask equipped with a stirrer, reflux condenser, thermometer and dropping funnel, was added 36.0 g of 25% aqueous sodium hydroxide solution, and the temperature was raised to 80°C. The above-obtained aqueous sodium salt solution of the starting material was added dropwise over 1 h while maintaining the temperature at 80°C, and stirring was continued for 1 h to obtain a reaction mixture.

[0047] Into a 250 mL four-necked flask equipped with a stirrer, reflux condenser, thermometer and dropping funnel, was added 20 mL of toluene and 31.2 g of 35% hydrochloric acid, and the above-obtained reaction mixture was added dropwise at room temperature over 1 h. The pH of the reaction system was adjusted to 2, and stirring was continued at room temperature for 30 min. The mixture was allowed to stand to separate into layers, and the resulting toluene solution of 4-chloro-2-fluoro-5-mercaptophenol was obtained in a yield of 96.6% as determined by HPLC analysis. A small amount of the solution was removed for analysis.

[0048] 1 H-NMR (300 MHz, CDC13) δ (ppm): 7.14 (d, J = 5.1 Hz, 1H), 7.01 (d, J = 4.4 Hz, 1H), 5.54 (bs, 1H), 3.81 (s, 1H). The melting point was 65.1°C.

[0049] Step 5, Preparation of 4-chloro-2-fluoro-5-(2,2,2-trifluoroethylthio)phenol;

[0050] Into a 1000 mL four-necked flask equipped with a stirrer, reflux condenser, thermometer and dropping funnel, was added 97.6 g of the toluene solution of 4-chloro-2-fluoro-5-mercaptophenol obtained in Step 4, and the toluene was distilled off under reduced pressure at 60°C. To the residue was added 65 mL of DMF and 92.5 g of 2,2,2-trifluoroethyl tosylate, and the temperature was raised to 80°C. Then, 58.3 g of 25% aqueous sodium hydroxide solution was added dropwise over 1 h, and the reaction was continued at 80°C for 2 h. Then, a mixture of 4.01 g of sodium formaldehyde sulfoxylate (Na-formaldoxime) and 17 mL of water was added dropwise over 1 h, and the reaction was continued at 80°C for 2 h.

[0051] The reaction mixture was cooled to 30-40°C, and 129 mL of toluene, 260 mL of water and 31.2 g of 25% aqueous sodium hydroxide solution were added thereto to adjust the pH of the reaction system to 12-13. The mixture was stirred at room temperature for 30 min, and allowed to stand to separate into layers. The resulting aqueous solution of the sodium salt of the product was washed with 129 mL of toluene to obtain a toluene solution of 4-chloro-2-fluoro-5-(2,2,2-trifluoroethylthio)phenol.

[0052] Into a 1000 mL four-necked flask equipped with a stirrer, a reflux condenser, a thermometer and a dropping funnel, was placed 260 mL of toluene and the above sodium salt aqueous solution. 81.2 g of 35% hydrochloric acid was added dropwise at room temperature, and the pH of the reaction system was adjusted to 4 or less. The mixture was stirred at room temperature for 30 minutes, and then allowed to stand to separate into layers. The organic layer was washed with 130 mL of water, and then distilled under reduced pressure to obtain 63.1 g of the product, 4-chloro-2-fluoro-5-(2,2,2-trifluoroethylthio)phenol, in a yield of 93.2%. The solution was directly used in the next step after distillation of a small amount of the solution for analysis.

[0053] 1 H-NMR (300 MHz, CDC13) δ (ppm): 7.27 (d, J = 8.7 Hz, 1H), 7.22 (d, J = 10.2 Hz, 1H), 5.15 (d, J = 3.9 Hz, 1H), 3.43 (q, J = 9.6 Hz, 2H).

[0054] Step 6, Preparation of (2-chloro-5-(4-chlorobutoxy)-4-fluorophenyl)-(2,2,2- trifluoroethyl) sulfide;

[0055] Into the reaction flask was added 15 mL of DMF, followed by 7.8 g of 4-chloro-2-fluoro-5-(2,2,2-trifluoroethylthio)phenol, 5.7 g of 1-chloro-4-bromobutane, 3.5 g of sodium carbonate, and 0.45 g of sodium iodide prepared in Step 5. The mixture was heated to 90°C and reacted for 8 hours. After confirming the completion of the reaction, the reaction mixture was cooled to 10°C and then 4.8 g of 25% sodium hydroxide aqueous solution and 30 mL of water were added. The reaction mixture was extracted with 30 mL x 2 of dichloromethane, and the combined organic phase was washed with 20 mL x 3 of brine. The organic phase was distilled under reduced pressure to obtain the product, (2-chloro-5-(4-chlorobutoxy)-4-fluorophenyl)-(2,2,2-trifluoroethyl) sulfide, in a yield of 90%.

[0056] 1 H-NMR (300 MHz, CDC13) δ (ppm): 7.53 (d, J = 8.0 Hz, 1H), 7.40 (d, J = 4.0 Hz, 1H), 4.03 (t, J = 7.2 Hz, 2H), 3.60 (t, J = 7.2 Hz, 2H), 3.41 (q, J = 10 Hz, 2H), 1.80-1.74 (m, 4H).

[0057] Step 7, Preparation of (4-(4-chloro-2-fluoro-5-((2,2,2-trifluoroethyl)methylsulfanyl)phenoxy)butyl)methyl sulfane (FW-10);

[0058] 20 mL of DMSO was added to the reaction flask, along with 7.0 g of (2-chloro-5-(4-chlorobutoxy)-4-fluorophenyl)-(2,2,2-trifluoroethyl) sulfide prepared in step 6 and 10.5 g of 20% sodium methanethiol aqueous solution. The mixture was heated to 90 °C and reacted for 12 h. After confirming the reaction was complete, the reaction system was cooled to 10 °C, 50 mL of water was added, and the reaction solution was extracted with dichloromethane (40 mL x 2). The organic phases were combined, washed with brine (30 mL x 3), and the organic phases were dissolved under reduced pressure to obtain the product (4-(4-chloro-2-fluoro-5-((2,2,2-trifluoroethyl)methylthion)phenoxy)butyl)methylthion (FW-10), with a yield of 81%.

[0059] 1 H-NMR(300MHz, CDCl3)δ(ppm):7.60(d,J=8.0Hz,1H),6.52(d,J=4.0Hz,1H),4.03(t,J=7.2Hz,2H ),3.31(q,J=10Hz,2H),2.60(t,J=7.2Hz,2H),2.10(s,3H),1.70-1.60(m,2H),1.55-1.50(m,2H).

[0060] Example 2

[0061] The compound FW-10 obtained in Example 1 was used as the technical grade pesticide to prepare suitable formulations, such as suspension concentrates, dispersible oil suspension concentrates, microcapsule suspension concentrates, emulsions, wettable powders, or water-dispersible granules. This example uses a dispersible oil suspension concentrate as an example, and suitable adjuvants were selected to prepare pesticides of different concentrations, the components of which are shown in Table 1.

[0062] Table 1

[0063]

[0064] In Table 1, CO / 40 (abbreviation for ethoxylated castor oil) and polyethylene glycol 400 are both emulsifiers; NS500LQ (abbreviation for nonionic hydroxyl polyethylene oxide block copolymer) is a dispersant; N,N-dimethylacetamide and N-butylpyrrolidone are cosolvents, and isopropanol is a solvent oil.

[0065] According to the formula in Table 1, add FW-10, N,N-dimethylacetamide, and N-butylpyrrolidone to the preparation vessel; stir for 45 min until the solution is clear, then add CO / 40, NS500LQ, and polyethylene glycol 400, and finally make up to 100% with isopropanol, stir for another 15 min, filter through a 200-mesh sieve, and dispense after passing the high-performance liquid chromatography (HPLC) analysis; the HPLC conditions include: column temperature 40℃±2℃; flow rate 1.5mL / min; mobile phase acetonitrile:phosphoric acid aqueous solution (0.1%) = 75:25; injection volume 10μL; wavelength 245nm; retention time approximately 8.5min.

[0066] Example 3

[0067] Indoor biological activity evaluation: Thysanoptera pests (taking thrips as an example).

[0068] Experimental targets: Thrips nymphs were used as test insects. They were collected from a chili field in Leizhou City, Guangdong Province. They were fed with cabbage leaves grown indoors without pesticides for 3 hours, and healthy second-instar nymphs were selected for use.

[0069] Experimental conditions: Temperature = 25℃~27℃; Relative humidity = 60%~80%; Photoperiod L / D = 14h / 10h.

[0070] Pharmaceutical preparation:

[0071] (1) Preparation of test group: Weigh a certain amount of preparation 1 to preparation 3 obtained in Example 2, add water to 100mL, prepare a 200mg / L drug solution, and then dilute with water to the required concentration. Select drug solutions with the concentration of active ingredient FW-10 of 0.5mg / L, 0.2mg / L, 0.1mg / L and 0.05mg / L as test drug solutions;

[0072] (2) Preparation of positive control group: The original drug of the control group is flonicamid. According to the preparation method of Example 2, FW-10 is replaced with an equal mass of flonicamid, and other components and preparation conditions remain unchanged to prepare 5%, 10% and 15% flonicamid preparations (referred to as control A1, control A2 and control A3). A certain amount of flonicamid preparation is weighed, and water is added to 100mL to prepare a 200mg / L drug solution. Then, the solution is diluted with water to the required concentration. The drug solutions with active ingredient concentrations of 0.5mg / L, 0.2mg / L, 0.1mg / L and 0.05mg / L are selected as test solutions.

[0073] Experimental method: Leaf tube film method.

[0074] (1) Fill each test drug solution of the test group and the control group into 1.5 mL Axygen centrifuge tubes, let stand for 4 hours, then pour out the drug solution and let it dry; repeat each concentration treatment 4 times, and cut the bottom of the Axygen centrifuge tube into a small hole with a diameter of about 3 mm to 5 mm with scissors;

[0075] (2) Take fresh cabbage leaves, wipe them clean, and use a punch to make leaf discs with a diameter of 1 cm for testing; immerse the pepper leaf discs in the test solutions of different concentrations for 10 seconds; place the treated leaf discs to air dry naturally at room temperature, and use small tweezers to separate the leaf discs along with the 1 cm diameter leaf discs. 2 Dry filter paper sheets of the appropriate size were placed into Axygen centrifuge tubes containing the corresponding test drug concentration, one sheet per tube, to form a leaf tube drug film;

[0076] (3) 40 healthy test worms of uniform size were inoculated into each Axygen centrifuge tube.

[0077] Investigation method: Three days after the drug was administered, the reaction symptoms and number of dead insects were investigated. The insects were judged to be dead if they could not move in a coordinated manner when touched with a brush. The mortality rate was calculated as (number of dead insects / total number of insects) * 100%. The experimental results are shown in Table 2.

[0078] Table 2

[0079]

[0080] Note: The data in Table 2 are mean ± standard deviation; different superscript letters between data in the same column indicate statistically significant differences (P<0.05), and the same superscript letters indicate no statistically significant differences (P>0.05).

[0081] As shown in Table 2, the FW-10 formulation of Example 2 of this invention exhibited good insecticidal effects against thrips. When formulations 1 to 3 (FW-10) and controls A1 to A3 (fluoxetine) of this invention were diluted to a high concentration (active ingredient = 0.5 mg / L) and applied, the mortality rate of test insects was 100% in all groups, with no difference between them. However, as the concentration of the active ingredient decreased, the mortality rate of test insects after application of the FW-10 formulation of this invention differed from that of the control fluoxetine formulations, and the difference was more pronounced at lower concentrations.

[0082] As can be seen from the data comparison in Table 2, at low concentrations, the activity of the control group (fluoxetine) is significantly different from that of the test group (FW-10). As a thrips insecticide, FW-10 of this invention is significantly more effective than flonicamid.

[0083] Example 4

[0084] Indoor biological activity evaluation: harmful mites (taking the citrus paronychia as an example).

[0085] Experimental targets: Citrus paronychia collected outdoors were placed in an indoor constant temperature incubator for breeding for more than 3 generations. Healthy, normally developed Citrus paronychia with consistent physiological conditions were selected as test mites. The breeding parameters were set as follows: temperature = 24℃~26℃, relative humidity = 60%~80%, photoperiod L / D = 14h / 10h. No chemical agents were used during the breeding period.

[0086] Indoor pharmaceutical preparation:

[0087] 1) Preparation of test group: Weigh a certain amount of preparation 1 to preparation 3 obtained in Example 2, add water to 100mL, prepare a 200mg / L drug solution, and then dilute with water to the required concentration. Select drug solutions with concentrations of active ingredient FW-10 of 0.5mg / L, 0.2mg / L, 0.1mg / L and 0.05mg / L as test drug solutions;

[0088] 2) Control group for preparing positive drugs: The original drug for the control group was pyridaben. According to the preparation method in Example 2, FW-10 was replaced with an equal mass of pyridaben, while other components and preparation conditions remained unchanged, to prepare 5%, 10%, and 15% pyridaben preparations (referred to as Control B1, Control B2, and Control B3). A certain amount of pyridaben preparation was weighed and added to 100 mL of water to prepare a 200 mg / L solution. Then, it was diluted with water to the required concentration. The solutions with the concentrations of the active ingredient pyridaben of 0.5 mg / L, 0.2 mg / L, 0.1 mg / L, and 0.05 mg / L were selected as the test solutions.

[0089] Experimental method: glass slide immersion method.

[0090] (1) Stick a 2cm×2cm double-sided tape to one end of the slide, leaving only 2mm to 3mm wide at the edge of the slide;

[0091] (2) Use a No. 0 brush to pick out healthy and uniform citrus red spider mites, and gently stick their backs onto a glass slide with double-sided tape. Do not stick the mouthparts, antennae and legs of the mites. 30 citrus red spider mites should be attached to each glass slide. Then place the glass slide in a large petri dish, put a wet cotton ball on it, cover it with a glass plate, and place it at 25°C. After 2 hours, examine it under a microscope to see if there are any dead or inactive individuals. If dead or inactive individuals are found, remove them and add healthy live mites before measuring again.

[0092] (3) Immerse the glass slides with the healthy citrus parsnips attached into the test solutions of the above test group and control group, gently shake for 5 seconds, take them out and place them on filter paper, and then gently absorb the excess solution around the mite and on the tape with a small piece of filter paper; repeat the treatment 4 times for each gradient, and after each concentration of agent is treated, put them back into the original large petri dish, and examine them under a binocular stereomicroscope after 48 hours at room temperature. Gently touch the limbs of the mite with a No. 0 brush. If the mite's legs do not move, it is considered dead. Record the number of dead mites and the number of live mites; the mortality rate = number of dead mites / total number of mites * 100%. The experimental results are shown in Table 3.

[0093] Table 3

[0094]

[0095] Note: The data in Table 3 are mean ± standard deviation; different superscript letters between data in the same column indicate statistically significant differences (P<0.05), and the same superscript letters indicate no statistically significant differences (P>0.05).

[0096] As shown in Table 3, the FW-10 formulation of Example 2 of the present invention exhibited good insecticidal effects against citrus paronychia. When formulations 1 to 3 (FW-10) and control groups B1 to B3 (pyridaben) of the present invention were diluted to a high concentration (active ingredient = 0.5 mg / L) and applied, the mortality rate of the tested mites was 100%, with no difference between the groups. However, as the concentration of the active ingredient decreased, the mortality rate of the tested mites after application of the FW-10 formulation of the present invention and the control group pyridaben showed differences, and the difference was more pronounced at lower concentrations.

[0097] As can be seen from the data comparison in Table 3, at low concentrations, the activity of the control group (pyridaben) is significantly different from that of the test group (FW-10). As a citrus acaricide, FW-10 of this invention is significantly more effective than pyridaben.

[0098] Example 5

[0099] Indoor biological activity evaluation: Lepidoptera pests (taking the rice stem borer as an example).

[0100] Experimental targets: Rice stem borers collected outdoors were reared with rice seeds in an artificial climate indoors for more than three generations. The rearing parameters were set as follows: temperature = 24℃~26℃, relative humidity = 60%~80%, photoperiod L / D = 14h / 10h. No chemical agents were used during the rearing period. Healthy, normally developed, and physiologically consistent third-instar rice stem borer larvae were selected as test insects.

[0101] Pharmaceutical preparation:

[0102] 1) Preparation of test group: Weigh a certain amount of preparation 1 to preparation 3 obtained in Example 2, add water to 100mL, prepare a 200mg / L drug solution, and then dilute with water to the required concentration. Select drug solutions with concentrations of active ingredient FW-10 of 0.5mg / L, 0.2mg / L, 0.1mg / L and 0.05mg / L as test drug solutions;

[0103] 2) Control group for preparing positive drug: The original drug for the control group was chlorpyrifos. According to the preparation method in Example 2, FW-10 was replaced with an equal mass of chlorpyrifos, while other components and preparation conditions remained unchanged, to prepare 5%, 10%, and 15% chlorpyrifos preparations (referred to as control C1, control C2, and control C3). A certain amount of chlorpyrifos preparation was weighed and added to 100 mL of water to prepare a 200 mg / L solution. Then, it was diluted with water to the required concentration. The solutions with the active ingredient chlorpyrifos concentrations of 0.5 mg / L, 0.2 mg / L, 0.1 mg / L, and 0.05 mg / L were selected as test solutions.

[0104] Test method: Immersion method;

[0105] Place water chestnut strips into the test solution of the test group and the control group, soak for 10 seconds, remove the water chestnut strips and dry them, and then place them in disposable petri dishes lined with filter paper. Place 10 rice stem borer larvae in each petri dish. Repeat each treatment group 4 times, and then transfer the petri dishes to an incubator for culture.

[0106] Investigation method: Five days after the drug was applied, the survival and mortality of the rice stem borer larvae were observed. The insects were judged to be dead if they could not move in a coordinated manner when touched with a brush. The mortality rate was calculated as (number of dead insects / total number of insects) * 100%. The experimental results are shown in Table 4.

[0107] Table 4

[0108]

[0109] Note: The data in Table 4 are mean ± standard deviation; different superscript letters between data in the same column indicate statistically significant differences (P<0.05), and the same superscript letters indicate no statistically significant differences (P>0.05).

[0110] As shown in Table 4, the FW-10 formulation of Example 2 of the present invention exhibited good insecticidal effects against the rice stem borer. When formulations 1 to 3 (FW-10) and controls C1 to C3 (insecticides) of the present invention were diluted to a high concentration (active ingredient = 0.5 mg / L) and applied, the mortality rate of the test insects was 100%, with no difference between the groups. However, as the concentration of the active ingredient decreased, the mortality rate of the test insects after application of the FW-10 formulation of the present invention differed from that of the control insecticides, and the difference was more pronounced at lower concentrations.

[0111] As can be seen from the data comparison in Table 4, at low concentrations, the activity of the control group (insecticide) is significantly different from that of the test group (FW-10). As an insecticide for rice stem borer, FW-10 of this invention is significantly more effective than insecticide.

[0112] Example 6

[0113] Indoor biological activity evaluation: Diptera pests (taking the citrus fruit fly as an example).

[0114] Experimental targets: Citrus fruit flies were collected from grapefruits and continuously bred in the laboratory for more than 3 generations. Adult citrus fruit flies from the same batch that emerged 10 days after hatching were selected as test insects. Before the experiment, they were starved with clean water for 24 hours. The breeding conditions were: temperature = 25℃±1℃, relative humidity = 75%±1%, and photoperiod L / D = 16h / 8h.

[0115] Pharmaceutical preparation:

[0116] (1) Preparation of test group: Weigh a certain amount of preparation 1 to preparation 3 prepared in Example 2, add 10% sucrose solution, add water to 100 mL, prepare a 200 mg / L solution, then dilute with water to the required concentration, and select solutions with concentrations of 0.5 mg / L, 0.2 mg / L, 0.1 mg / L and 0.05 mg / L of active ingredient FW-10 as test solutions;

[0117] (2) Preparation of positive control group: The original drug of the control group is avermectin. According to the preparation method of Example 2, FW-10 is replaced with an equal mass of avermectin, and other components and preparation conditions remain unchanged to prepare 5%, 10% and 15% avermectin preparations (referred to as control D1, control D2 and control D3). A certain amount of avermectin preparation is weighed, 10% sucrose solution is added, and then water is added to 100mL to prepare a 200mg / L drug solution. Then, the solution is diluted with water to the required concentration. The drug solutions with the concentration of the active ingredient avermectin of 0.5mg / L, 0.2mg / L, 0.1mg / L and 0.05mg / L are selected as test drug solutions.

[0118] Experimental method: Liquid feeding method;

[0119] Immerse a sponge (5cm×5cm×5cm) in the test solution of the test group and the control group for 30 seconds, then remove it and let it stand naturally until the test solution stops dripping. Place the sponge in a 1000mL conical flask. Introduce 30 adult citrus fruit flies (half male and half female) into the conical flask, seal the flask with a perforated cap, and place it in an artificial climate chamber. Continue to raise the flies under the conditions of temperature = 25℃±1℃, relative humidity = 75%±1%, and photoperiod L / D = 16h / 8h. Each concentration was replicated 4 times.

[0120] Investigation method: 48 hours after drug application, the survival and mortality of adult citrus fruit flies were observed. The insects were judged to be dead if they could not move in a coordinated manner when touched with a brush. The mortality rate was calculated as (number of dead insects / total number of insects) * 100%. The experimental results are shown in Table 5.

[0121] Table 5

[0122]

[0123] Note: The data in Table 5 are mean ± standard deviation; different superscript letters between data in the same column indicate statistically significant differences (P<0.05), and the same superscript letters indicate no statistically significant differences (P>0.05).

[0124] As shown in Table 5, the FW-10 formulation of Example 2 of the present invention exhibited good insecticidal effects against fruit flies. When formulations 1 to 3 (FW-10) and controls D1 to D3 (avermectin) of the present invention were diluted to a high concentration (active ingredient = 0.5 mg / L) and applied, the mortality rate of test insects was 100% in all groups, with no difference between them. However, as the concentration of the active ingredient decreased, the mortality rate of test insects after application of the FW-10 formulation of the present invention differed from that of the control avermectin, and the difference was more pronounced at lower concentrations.

[0125] As can be seen from the data comparison in Table 5, at low concentrations, the activity of the control group (avermectin) is significantly different from that of the test group (FW-10). As a fruit fly insecticide, FW-10 of this invention is significantly more effective than abamectin.

[0126] Example 7

[0127] Pest repellency test: Lepidoptera pests (taking cotton bollworm as an example);

[0128] Pharmaceutical preparation:

[0129] (1) Preparation of test group: Weigh a certain amount of preparation 1 to preparation 3 obtained in Example 2, add water to 100mL, prepare a 200mg / L drug solution, and then dilute with water to the required concentration. Select drug solutions with the concentration of active ingredient FW-10 of 0.5mg / L, 0.2mg / L, 0.1mg / L and 0.05mg / L as test drug solutions.

[0130] (2) Preparation of positive control group: The original drug of the control group is emamectin benzoate. According to the preparation method of Example 2, FW-10 is replaced with an equal mass of emamectin benzoate, and other components and preparation conditions remain unchanged to prepare 5%, 10% and 15% emamectin benzoate preparations (referred to as control E1, control E2 and control E3). A certain amount of emamectin benzoate is weighed and added to 100 mL of water to prepare a 200 mg / L drug solution. Then, it is diluted with water to the required concentration. Drug solutions with the concentration of the active ingredient emamectin benzoate of 0.5 mg / L, 0.2 mg / L, 0.1 mg / L and 0.05 mg / L are selected as test drug solutions.

[0131] Experimental targets: Cotton bollworms collected outdoors were reared in an indoor constant-temperature incubator with parameters set as follows: temperature = 24℃~26℃, relative humidity = 60%~80%, and photoperiod L / D = 14h / 10h. The adults were reared until they emerged, mated, and laid eggs. The resulting F1 generation larvae were used for subsequent experiments. No chemical agents were used during the rearing period. Second-instar larvae with normal development and consistent physiological conditions were selected for the experiment and starved for 12 hours before their food intake was measured.

[0132] Test method:

[0133] (1) Place cotton plants in the bud stage in a breathable container and spray 100 mL of the test solution of each test group and control group evenly on the leaf surface of the cotton plants. Set 10 replicates for each concentration.

[0134] (2) The treated cotton plants were placed in a cotton bollworm incubator, with one cotton bollworm in each incubator, and cotton plants sprayed with water were used as a control group.

[0135] (3) Take out cotton plants every day, use sulfuric acid paper to print feeding patches and calculate the feeding area; if cotton bollworms die, new cotton bollworms should be added in time and observed for 24 hours. After the observation period, measure the total feeding area and calculate the repellency rate of FW-10 and abamectin against cotton bollworms. The results are shown in Table 6.

[0136] Calculate the avoidance rate = (S 对照 -S 处理 ) / S 对照 ×100%, S 处理 and S 对照 The feeding area is shown in the experimental drug spray treatment group and the water control group, respectively.

[0137] Table 6

[0138]

[0139] Note: The data in Table 6 are mean ± standard deviation; different superscript letters between data in the same column indicate statistically significant differences (P<0.05), and the same superscript letters indicate no statistically significant differences (P>0.05).

[0140] As shown in Table 6, the FW-10 formulation of Example 2 of the present invention exhibited good repellency against cotton bollworms. When formulations 1 to 3 (FW-10) and controls E1 to E3 (emamectin benzoate) of the present invention were diluted to a high concentration (active ingredient = 0.5 mg / L) and applied, the repellency rate against cotton bollworms was 100% after 24 hours, with no difference between the groups. However, as the concentration of the active ingredient decreased, the repellency rate of cotton bollworms after application of the FW-10 formulation of the present invention differed from that of the controls using emamectin benzoate, and the difference was more pronounced at lower concentrations.

[0141] As can be seen from the data comparison in Table 6, at low concentrations, the activity of the control group (emamectin benzoate) is significantly different from that of the test group (FW-10). As a cotton bollworm repellent, FW-10 of this invention is significantly more effective than emamectin benzoate.

[0142] Example 8

[0143] Mite repellency test: taking the citrus parsnipus mite as an example;

[0144] Pharmaceutical preparation:

[0145] (1) Preparation of test group: Weigh a certain amount of preparation 1 to preparation 3 obtained in Example 2, add water to 100mL, prepare a 200mg / L drug solution, and then dilute with water to the required concentration. Select drug solutions with the concentration of active ingredient FW-10 of 0.5mg / L, 0.2mg / L, 0.1mg / L and 0.05mg / L as test drug solutions.

[0146] (2) Preparation of positive control drugs: The original drug for the control group was flufenoxuron. According to the preparation method in Example 2, FW-10 was replaced with an equal mass of flufenoxuron, while other components and preparation conditions remained unchanged. Flufenoxuron preparations of 5%, 10%, and 15% were prepared (referred to as control F1, control F2, and control F3). A certain amount of flufenoxuron preparation was weighed and added to 100 mL of water to prepare a 200 mg / L solution. Then, the solution was diluted with water to the required concentration. The solutions with concentrations of the active ingredient flufenoxuron of 0.5 mg / L, 0.2 mg / L, 0.1 mg / L, and 0.05 mg / L were selected as test solutions.

[0147] Experimental targets: Citrus paronychia collected outdoors were reared in an indoor constant-temperature incubator for at least three generations. Healthy, normally developed citrus paronychia with consistent physiological conditions were selected as test mites. The rearing parameters were set as follows: temperature = 24℃~26℃, relative humidity = 60%~80%, photoperiod L / D = 14h / 10h. No chemical agents were used during the rearing period. Second-instar larvae with normal development and consistent physiological conditions were selected for the experiment and starved for 12 hours before their feeding amount was measured.

[0148] Experimental Method: Tender leaves were taken from healthy citrus plants; leaf discs with a diameter of 3 cm were punched using a puncher, and the leaf discs were soaked in the test solution for 10 minutes and then air-dried. Two leaf discs soaked in water (control) and two leaf discs soaked in the test solution were placed in an artificial incubator. The two leaf discs treated with the test solution and the two leaf discs soaked in water (control) were arranged in a cross shape, with a spacing of about 2 cm between each leaf disc. One citrus paronychia mite was introduced into each incubator; 10 replicates were set up for each concentration.

[0149] After the 24-hour experiment, the remaining area of ​​the leaf disc was measured, and the feeding area was calculated. The feeding area = initial leaf disc area - remaining leaf disc area. The avoidance rate was calculated as (S...). 对照 -S 处理 ) / S 对照 ×100%, S 处理 and S 对照 The feeding area of ​​the experimental drug solution immersion group and the water control group are shown in Table 7.

[0150] Table 7

[0151]

[0152] Note: The data in Table 7 are mean ± standard deviation; different superscript letters between data in the same column indicate statistically significant differences (P<0.05), and the same superscript letters indicate no statistically significant differences (P>0.05).

[0153] As shown in Table 7, the FW-10 formulation of Example 2 of the present invention exhibited good repellency against the citrus paronychia. When formulations 1 to 3 (FW-10) and comparative formulations F1 to F3 (flufenoxam) of the present invention were diluted to a high concentration (active ingredient = 0.5 mg / L) and applied, the repellency rate against citrus paronychia was 100% after 24 hours, with no difference between the groups. However, as the concentration of the active ingredient decreased, the repellency rate of citrus paronychia after application of the FW-10 formulation of the present invention and the comparative flufenoxam showed differences, and the difference was more pronounced at lower concentrations.

[0154] As can be seen from the data comparison in Table 7, at low concentrations, the activity of the control group (Citrus parvovirus) was significantly different from that of the test group (FW-10). The FW-10 of this invention, as a repellent for Citrus parvovirus, is significantly more effective than flufenoxuron.

[0155] Example 9

[0156] Although the compound prepared in Example 1 exhibits excellent biological activity, its biological safety is low in practical applications because thioether compounds have extremely high environmental toxicity, especially to soil organisms, aquatic organisms, and other beneficial insects.

[0157] To address the aforementioned issues, the present invention incorporates a biosafety agent into the pesticide composition prepared in Example 2, which improves the biosafety of the pesticide composition without reducing its biological activity.

[0158] The preparation method of biosafety agents includes the following steps:

[0159] S1, preparation of core raw materials;

[0160] (1) Dissolve GST enzyme powder in PBS buffer (0.1M, pH=7.4) pre-cooled to 4℃ to obtain a solution with a GST enzyme concentration of about 10mg / mL; then add trehalose to a final concentration of 5% (w / v) to obtain a GST enzyme solution.

[0161] (2) N-acetylcysteine ​​was dissolved in PBS buffer (0.1M, pH=7.4) to obtain a NAC solution of about 50 mg / mL;

[0162] (3) Heat deionized water to above 70°C and keep it warm. Slowly add pectin powder while stirring. Continue stirring and heating until the pectin is fully dissolved. Continue stirring and cool to room temperature to obtain a pectin solution.

[0163] (4) According to the volume ratio of NAC solution: pectin solution = 1:9, NAC solution is slowly added to pectin solution under stirring conditions, and NAC / pectin solution is obtained after thorough mixing;

[0164] (5) Chitosan was dispersed in deionized water to obtain a chitosan dispersion with a mass concentration of about 5 mg / mL. 1% acetic acid solution was added and magnetically stirred until completely dissolved to obtain a chitosan-acetic acid solution.

[0165] S2, Prepare hydrogel microspheres encapsulating NAC;

[0166] The NAC / pectin solution was slowly added dropwise to the chitosan-acetic acid solution (mass ratio of the two is about 1:1) under high-speed stirring at about 500 rpm. After the addition was completed, stirring was continued for about 30 min to obtain a suspension. The suspension was centrifuged (5000 rpm, 10 min), the supernatant was discarded and the precipitate was collected to form hydrogel microspheres encapsulating NAC.

[0167] S3, Prepare hydrogel microspheres loaded with GST enzyme-NAC;

[0168] Following a GST enzyme:NAC mass ratio of 1:5, the NAC-encapsulated hydrogel microspheres prepared in step S2 were resuspended in the GST enzyme solution prepared in step S1. The microspheres were then slowly incubated at 4°C and 100 rpm with gentle shaking for 12 hours to allow the GST enzyme to diffuse into the hydrogel network. After centrifugation (5000 rpm, 10 min), the supernatant was discarded and the precipitate was collected to obtain GST enzyme-NAC-loaded hydrogel microspheres.

[0169] S4, Preparation of nanocapsule shells

[0170] The hydrogel microspheres (approximately 100 mg) loaded with GST enzyme-NAC prepared in S3 were dispersed in 40 mL of anhydrous ethanol, 1 mL of ammonia was added, and the mixture was stirred for 10 minutes. Then, 0.2 mL of tetraethyl orthosilicate (TEOS) was slowly added dropwise, and the reaction was continued for 6 hours. TEOS underwent hydrolysis and condensation under ammonia catalysis, forming a dense porous silica shell on the surface of the hydrogel microspheres. The mixture was centrifuged (5000 rpm, 5 min), and the supernatant was discarded to collect the precipitate, thus obtaining silica-encapsulated microcapsules.

[0171] S5, with a composite coating on the outer casing.

[0172] The microcapsules prepared by S4 were dispersed in 50 mL of Tris-HCl buffer (10 mM, pH=8.5); 50 mg of dopamine hydrochloride was added, and the mixture was stirred at room temperature in the dark for 12 hours. Dopamine self-polymerized on the surface of the silica shell to form a polydopamine layer (PDA). The mixture was centrifuged (5000 rpm, 5 min), and the supernatant was discarded to collect the precipitate, obtaining microcapsules with a PDA coating. The microcapsules were then dispersed in 20 mL of MES buffer (0.1 M, pH=5.5), and 20 mg of chitin oligosaccharide (COS) and 40 mg of coupling agent 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) were added. The mixture was reacted at room temperature for about 4 hours to attach the chitin oligosaccharide to the polydopamine layer using a coupling reaction. The mixture was centrifuged (5000 rpm, 5 min), and the supernatant was discarded to collect the precipitate, obtaining microcapsules with a shell covered by a composite coating (PDA / COS).

[0173] S6, Post-processing

[0174] The microcapsules prepared by S5 were centrifuged and washed to remove unreacted components, and then freeze-dried under vacuum to obtain nanoparticles, which became the biosafety agent.

[0175] Example 10

[0176] The biosafety agent prepared in Example 9 was subjected to biosafety verification. After adding the biosafety agent to the pesticide composition, it was verified whether it had an inhibitory effect on the control and repellency of crop pests or mites.

[0177] 1. Experimental Grouping

[0178] FW-10 group: This is a single-agent pesticide group. Formulation 1 prepared in Example 2 was selected and the test solution with an effective concentration of FW-10 of 0.05 mg / L was prepared.

[0179] FW-10+SF-L group: This is a pesticide + low-dose biosafety agent group. The amount of biosafety agent prepared in Example 9 is 0.5 times the mass of FW-10 in the pesticide single agent group. It is added to formulation 1 prepared in Example 2 and mixed. The mixture is then sand-milled in the liquid phase and diluted with water to prepare a test solution with an effective concentration of 0.05 mg / L of FW-10.

[0180] FW-10+SF-M group: This is a pesticide + medium-dose biosafety agent group. The amount of biosafety agent prepared in Example 9 is 1 times the mass of FW-10 in the pesticide single agent group. It is added to formulation 1 prepared in Example 2 and mixed. The mixture is then sand-milled in the liquid phase and diluted with water to prepare a test solution with an effective concentration of 0.05 mg / L of FW-10.

[0181] FW-10+SF-H group: This is a pesticide + high-dose biosafety agent group. The amount of biosafety agent prepared in Example 9 is 1.5 times the mass of FW-10 in the pesticide single agent group. It is added to formulation 1 prepared in Example 2 and mixed. The mixture is then sand-milled in the liquid phase and diluted with water to prepare a test solution with an effective concentration of 0.05 mg / L of FW-10.

[0182] SF group: This is the single-agent group of biosafety agents. The biosafety agent powder prepared in Example 9 is mixed with dispersant Morwet D-425, wetting agent Tween 20, and antifreeze glycerol. The mixture is then sand-milled in an aqueous phase and diluted with water to prepare a test solution with an effective concentration of 0.15 mg / L of biosafety agent.

[0183] 2. The experimental procedure was the same as in Examples 3 to 8, except that the experimental drug solution used was the same as the grouped experimental drug solution described above. Other aspects, such as the experimental target, experimental method, and calculation method, were the same as in Examples 3 to 8. The test results are shown in Table 8 below.

[0184] Table 8

[0185]

[0186] Note: The data in Table 8 are mean ± standard deviation; different superscript letters between data in the same column indicate statistically significant differences (P<0.05), and the same superscript letters indicate no statistically significant differences (P>0.05).

[0187] As shown in Table 8, there were no significant differences in the control efficacy against each target organism among all treatment groups containing pesticides (i.e., pesticide-only groups and pesticide + low / medium / high dose biosafety agent groups). The biosafety agent-only groups showed very weak control efficacy against each target organism. This indicates that the addition of biosafety agents did not significantly affect the insecticidal, acaricidal, or repellent activities of the pesticides themselves, and their efficacy was comparable to that of pesticides used alone.

[0188] Example 11

[0189] Non-target biological toxicity tests were conducted on the biosafety agent prepared in Example 9. After adding the biosafety agent to the pesticide composition, the test was conducted to see if its toxicity to non-target organisms was reduced; non-target organisms included soil organisms (earthworms as an example), natural enemy insects (ladybugs as an example), pollinating insects (honeybees as an example), and aquatic organisms (Daphnia magna as an example).

[0190] 1. Experimental grouping;

[0191] FW-10 group: This is a single-agent pesticide group. Formulation 1 prepared in Example 2 was selected and a test solution with an effective concentration of 50 mg / L for FW-10 was prepared.

[0192] FW-10+SF-L group: This is a pesticide + low-dose biosafety agent group. The amount of biosafety agent prepared in Example 9 is 0.5 times the mass of FW-10 in the pesticide single agent group. It is added to formulation 1 prepared in Example 2 and mixed. The mixture is then sand-milled in the liquid phase and diluted with water to prepare a test solution with an effective concentration of 50 mg / L of FW-10.

[0193] FW-10+SF-M group: This is a pesticide + medium-dose biosafety agent group. The amount of biosafety agent prepared in Example 9 is 1 times the mass of FW-10 in the pesticide single agent group. It is added to formulation 1 prepared in Example 2 and mixed. The mixture is then sand-milled in the liquid phase and diluted with water to prepare a test solution with an effective concentration of 50 mg / L of FW-10.

[0194] FW-10+SF-H group: This is a pesticide + high-dose biosafety agent group. The amount of biosafety agent prepared in Example 9 is 1.5 times the mass of FW-10 in the pesticide single agent group. It is added to formulation 1 prepared in Example 2 and mixed. The mixture is then sand-milled in the liquid phase and diluted with water to prepare a test solution with an effective concentration of 50 mg / L of FW-10.

[0195] SF group: This is the single-agent group of biosafety agents. The biosafety agent powder prepared in Example 9 is mixed with dispersant Morwet D-425, wetting agent Tween 20, and antifreeze glycerol. The mixture is then sand-milled in an aqueous phase and diluted with water to prepare a test solution with an effective concentration of 50 mg / L of biosafety agent.

[0196] 2. Toxicity testing on soil organisms;

[0197] (1) Test target: healthy adult earthworms with a weight of 400mg±50mg and clearly visible clitellum.

[0198] (2) Artificial soil: 10wt% peat moss powder, 20wt% kaolin, and 70wt% industrial quartz sand powder were mixed, and calcium carbonate particles were added to adjust the pH to 6.0±0.5; according to the effective concentration of FW-10 of 20mg / kg, a certain amount of the test solution of FW-10 group, FW-10+SF-L group, FW-10+SF-M group, and FW-10+SF-H group were mixed evenly with artificial soil; according to the concentration of biosafety agent = 50mg / kg, the SF group was mixed evenly with artificial soil, and a blank group with only artificial soil as control was set up; deionized water was added to each group of artificial soil to make the soil moisture 35%~40%.

[0199] (3) Experimental procedure: The treated artificial soil (wet weight = 800g) was placed into a 1L culture tank and 10 earthworms were placed in each culture tank; each group was repeated 5 times; the mouth of the tank was covered with a perforated plastic film to keep it ventilated while preventing excessive evaporation of water; the culture tank was placed in a culture box at 20℃±2℃ in continuous darkness; the soil was weighed and water was added weekly to maintain constant soil moisture; on the 7th day after the start of the experiment, 5g of crushed cooked oat flakes were added to the surface of the soil in each culture tank as food.

[0200] (4) Investigation method: After 14 days of cultivation, earthworms were taken out of the artificial soil and cleaned. They were dried with filter paper and their tails were gently touched with a glass rod. Those without any reaction were considered dead. Those with broken bodies, liquefaction, or discoloration were also considered dead. The mortality rate of earthworms in each group and the corrected mortality rate of earthworms were calculated. The results are shown in Table 9.

[0201]

[0202]

[0203] D 活蚯蚓 D 死蚯蚓 The numbers represent the initial number of live earthworms in each group and the number of earthworms that died at the end of the experiment.

[0204] QMOR 试验组 QMOR 空白组 The mortality rates of earthworms in the experimental group and the control group were respectively.

[0205] 3. Toxicity testing on natural enemy insects;

[0206] (1) Experimental targets: healthy and active ladybugs (third instar larvae) of uniform size and age.

[0207] (2) Preparation of residual drug film: Clean the inner wall and bottom of the culture dish (9cm diameter), and spray the test drug solutions of FW-10 group, FW-10+SF-L group, FW-10+SF-M group, FW-10+SF-H group and SF group evenly onto the inner wall and bottom of the culture dish, and use water as a blank control; ventilate and air dry naturally to form a uniform drug film.

[0208] (3) Experimental procedure: Ladybugs were gently transferred to culture dishes with prepared drug film. Ten ladybugs were placed in each culture dish, and each group was repeated 5 times. Sufficient live aphids that were not contaminated by pesticides were added to each culture dish as food. To prevent ladybugs from escaping, the culture dish lid was covered with fine mesh or perforated to ensure ventilation. The culture dishes were placed in a climate chamber at 25℃±1℃, 70%RH, and photoperiod L / D=16h / 8h for 72h.

[0209] (4) Investigation method: After 72 hours of cultivation, ladybugs were gently touched with a brush. Those that did not react at all were recorded as dead; those whose bodies were shriveled, blackened or broken were also recorded as dead; the mortality rate of ladybugs in each group and the corrected mortality rate of ladybugs were calculated, and the results are shown in Table 9.

[0210]

[0211]

[0212] D 活瓢虫 D 死瓢虫 The numbers represent the initial number of live ladybugs in each group and the number of ladybugs that died at the end of the experiment.

[0213] PMOR 试验组 PMOR 空白组 The mortality rates of ladybugs in the experimental group and the control group were respectively.

[0214] 4. Toxicity testing on pollinating insects;

[0215] (1) Test targets: Select healthy and active bees and acclimatize them for 3 hours in laboratory conditions (28℃~30℃) with 50% (w / v) sucrose water. Before the test, starve all the bees in the dark, at 25℃±1℃ and 70%RH for 2 hours to empty their honey sacs.

[0216] (2) Preparation of feeding solution: The test drug solution (100 mL) of FW-10 group, FW-10+SF-L group, FW-10+SF-M group, FW-10+SF-H group and SF group were mixed with 50% (w / v) sucrose water (100 mL) to prepare the feeding solution, and the 50% (w / v) sucrose water was used as the blank control.

[0217] (3) Experimental procedure: Bees were individually placed in special bee feeding tubes, and a small centrifuge tube containing 100 μL of the corresponding feeding solution was placed above each feeding tube; the bees fed themselves through the opening at the bottom of the centrifuge tube. There were 10 bees in each group, and each group was repeated 5 times; after feeding, the feeding tubes were removed and replaced with feeding tubes containing 50% (w / v) sucrose water as maintenance food, and the bees were put back into the incubator and observed for 48 hours.

[0218] (4) Investigation method: After 48 hours of cultivation, if the bees are completely still and do not react when their antennae are touched with tweezers, they are considered dead. The mortality rate of each group of bees and the corrected mortality rate of bees are calculated. The results are shown in Table 9.

[0219]

[0220]

[0221] D 活蜜蜂 D 死蜜蜂 The numbers represent the initial number of live bees in each group and the number of bees that died at the end of the experiment.

[0222] MMOR 试验组 MMOR 空白组 The mortality rates of bees in the experimental group and the control group were respectively.

[0223] 5. Toxicity testing on aquatic organisms;

[0224] (1) Experimental target: Select healthy large Daphnia clone populations for culture in the laboratory and select large Daphnia juveniles with consistent physiological state.

[0225] (2) Preparation of test solutions: The test solutions of FW-10 group, FW-10+SF-L group, FW-10+SF-M group and FW-10+SF-H group were diluted with water to the effective concentration of FW-10 = 1.0 mg / L. The test solution of SF group was diluted with water to the effective concentration of biosafety agent = 1.0 mg / L. Water was used as the blank control.

[0226] (3) Experimental procedure: Take a 100 mL beaker, add 50 mL of test solution, randomly put 10 juvenile Daphnia into each beaker, repeat 5 times per group, and then place the beaker in a completely dark incubator at 20℃±1℃ to prevent photosynthesis interference and expose it continuously for 48 hours; do not feed during the experiment.

[0227] (4) Investigation method: After 48 hours of cultivation, gently touch the abdomen of the juvenile daphnia with a pipette. If it cannot swim within 15 seconds, it means that it has lost its swimming ability. Check and record the number of juvenile daphnia that have lost their swimming ability in each beaker. Calculate the activity inhibition rate and the corrected activity inhibition rate of each group of juvenile daphnia. The results are shown in Table 9.

[0228]

[0229]

[0230] D 活幼溞 D 死幼溞 These represent the initial number of Daphnia juveniles in each group and the number of Daphnia juveniles that lost their ability to swim after the experiment.

[0231] IR 试验组 IR 空白组 The activity inhibition rates of the young Daphnia in the experimental group and the control group are respectively.

[0232] Table 9

[0233]

[0234] Note: The data in Table 9 are mean ± standard deviation; different superscript letters between data in the same column indicate statistically significant differences (P<0.05), and the same superscript letters indicate no statistically significant differences (P>0.05).

[0235] Based on the toxicity test data of soil organisms (earthworms) in Table 9, it can be seen that the mean corrected mortality rate of earthworms in the pesticide single-agent group was higher than 80%, which is at the level of toxicity and has strong toxicity to natural enemies; the earthworm corrected mortality rate in the biosafety agent single-agent group was 0, proving that it is safe for soil organisms; the mean corrected mortality rate of earthworms in the pesticide + low / medium / high dose biosafety agent groups showed different degrees of decrease. With the increase of biosafety agent dosage, the mean corrected mortality rate of earthworms decreased significantly, indicating that the biosafety agent can significantly and dose-dependently reduce the acute toxicity of pesticide FW-10 to earthworms and effectively protect soil organisms.

[0236] Based on the toxicity test data of natural enemy insects (ladybugs) in Table 9, it can be seen that the average corrected mortality rate of ladybugs in the pesticide single-agent group was higher than 80%, which is at the level of toxicity and has strong toxicity to natural enemies. The corrected mortality rate of ladybugs in the biosafety agent single-agent group was 0, proving that it is harmless and safe to natural enemy insects. The average corrected mortality rate of ladybugs in the pesticide + low / medium / high dose biosafety agent groups showed different degrees of decrease. With the increase of biosafety agent dosage, the average corrected mortality rate of ladybugs decreased significantly, indicating that the biosafety agent can significantly and dose-dependently reduce the acute toxicity of pesticide FW-10 to ladybugs and effectively protect natural enemy insects.

[0237] Based on the toxicity test data for pollinating insects (honeybees) in Table 9, it can be seen that the mean corrected mortality rate of honeybees in the pesticide single-agent group is higher than 90%, which is at the high toxicity level and poses a great risk to honeybees; the mean corrected mortality rate of honeybees in the biosafety agent single-agent group is less than 10%, proving that it is safe for honeybees; the mean corrected mortality rate of honeybees in the pesticide + low / medium / high dose biosafety agent groups shows different degrees of decrease. As the dose of biosafety agent increases, the mean corrected mortality rate of honeybees decreases significantly, indicating that the biosafety agent can significantly and dose-dependently reduce the acute oral toxicity of pesticide FW-10 to honeybees, effectively protect pollinating insects, and greatly reduce the environmental risks of pesticide use.

[0238] Based on the toxicity test data of aquatic organisms (Daphnia magna) in Table 9, it can be seen that the average corrected activity inhibition rate of the pesticide single-agent group is close to 90%, which is in the high toxicity range and poses a serious threat to the aquatic ecosystem; the average corrected activity inhibition rate of the biosafety agent single-agent group is less than 10%, proving that it is safe for Daphnia magna itself; the average corrected activity inhibition rate of the pesticide + low / medium / high dose biosafety agent groups shows different degrees of decrease. As the dose of biosafety agent increases, the average corrected activity inhibition rate decreases significantly, indicating that the biosafety agent can significantly and dose-dependently reduce the acute toxicity of pesticide FW-10 to Daphnia magna.

[0239] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

Claims

1. A pesticide composition, characterized in that, The pesticide composition comprises a trifluoroethyl phenyl sulfide compound or a salt thereof, and an agriculturally acceptable formulation carrier or adjuvant; the active ingredient of the pesticide composition comprises the trifluoroethyl phenyl sulfide compound or a salt thereof, wherein the trifluoroethyl phenyl sulfide compound has the following structure: ； The pesticide composition also includes a biosafety agent, which is in the form of a core-shell microcapsule with a core of GST enzyme-NAC loading, a middle layer of nanocapsule shell, and an outer layer of composite coating; GST enzyme is glutathione thiol transferase, and NAC is N-acetylcysteine.

2. The pesticide composition according to claim 1, characterized in that, The pesticide composition is used as an insecticide to control crop pests and / or mites; or, the pesticide composition is used as a repellent to repel crop pests and / or mites.

3. The pesticide composition according to claim 2, characterized in that, The crop pests are at least one of the following: Thysanoptera pests, Diptera pests, and Lepidoptera pests.

4. The pesticide composition according to claim 1, characterized in that, Based on the mass ratio, the ratio of biosafety agent to trifluoroethyl phenyl sulfide compound or its salt is 0.5~1.5:

1.

5. The pesticide composition according to claim 1, characterized in that, The preparation method of the biosafety agent includes the following steps: S1. GST enzyme solution and NAC solution were prepared using PBS buffer as solvent. Pectin was dissolved in hot water and cooled to room temperature. It was then mixed with NAC solution to prepare NAC / pectin solution. S2, through ion cross-linking, chitosan-acetic acid solution is mixed with NAC / pectin solution to form hydrogel microspheres encapsulating NAC; S3. GST enzyme solution is loaded into hydrogel microspheres prepared in S2 by diffusion method to obtain hydrogel microspheres loaded with GST enzyme-NAC. S4. In an ethanol and ammonia system, tetraethyl orthosilicate is used to perform a hydrolysis-condensation reaction on the surface of the hydrogel microspheres prepared in S3 to form a silica shell and obtain microcapsules. S5, in Tris-HCl buffer, a polydopamine layer is formed on the surface of a silica shell by dopamine self-polymerization; chitin oligosaccharide is attached to the polydopamine layer by a coupling reaction; S6 involves centrifugation and washing to remove unreacted components, followed by vacuum freeze-drying to obtain dried nanoparticles, which serve as the biosafety agent.

6. The pesticide composition according to claim 5, characterized in that, The GST enzyme solution also includes trehalose, with a final concentration of 1% to 5%.

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