Use of ricinine, rodenticide, and preparation method therefor and use thereof

Abstract

A use of ricinine in preparation of a rodenticide. The effective concentration of ricinine is 0.45-1.80%. A rodenticide comprises ricinine, a base bait, a stabilizing agent, and a bittering agent. The feeding coefficient of the prepared rodenticide is 0.52 or above. Laboratory tests show that the rodent mortality rate reaches 100% on the 9th day, and field tests show that the rodent mortality rate reaches 95.17% on the 21st day. In addition, the rodenticide meets safety requirements for usage, storage and transportation, and complies with stability standards.

Description

Application of ricinine, a rodenticide, and its preparation method and application Technical Field

[0001] The present invention relates to the field of biotechnology, in particular to the application of ricinine, a rodenticide and a preparation method and application thereof. Background Art

[0002] Most of the current rodent control agents are anticoagulant rodenticides, either 4-hydroxycoumarins or indandiones, such as coumatetralyl, warfarin, diphacin, bromadiolone, bromodiolone, and daclofenac. The principle is that they can competitively inhibit vitamin K in mice and hinder the synthesis of prothrombin, thereby leading to fatal internal bleeding and death.

[0003] In addition, a small number of fumigant rodenticides used in specific places such as small enclosed environments, such as chloropicrin, aluminum phosphide, and acute rodenticide fluoroacetamide, have been banned due to their high toxicity.

[0004] With the widespread use of chemical rodenticides, feeding resistance is a common occurrence, reducing the effectiveness of anticoagulant rodenticides. With increasing environmental safety requirements and technological advancements, biorodenticides are a key research and development priority. For example, botulinum toxin type C has demonstrated excellent efficacy against plateau pikas and zokors in field trials and has been registered. However, the availability of biorodenticides is extremely limited. Summary of the Invention

[0005] The purpose of the present invention is to provide an application of ricinine. The rodenticide prepared from ricinine has a significant rodenticide effect and is also green, easily degradable without residue, highly ecologically compatible and has a simple preparation process.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The present invention provides the use of ricinine in preparing rodenticide.

[0008] Preferably, the effective concentration of ricinine is 0.45-1.80%.

[0009] The present invention also provides a rodenticide comprising the following components in parts by mass: 0.45 to 1.80 parts of ricinine, 97 to 99 parts of base bait, 0.1 to 0.3 parts of stabilizer, and 0.002 to 0.02 parts of bittering agent.

[0010] Preferably, the base bait is one or more of corn flour, milk powder, peanut cake, and vegetable oil; the stabilizer is butylated hydroxytoluene; and the bittering agent is denatonium benzoate.

[0011] The present invention also provides a method for preparing the rodenticide, comprising the following steps:

[0012] (1) mixing ricin, base bait, stabilizer, and bittering agent to obtain a mixture;

[0013] (2) granulating the mixture and drying it to obtain a rodenticide.

[0014] Preferably, the drying method is heating in a dryer; the drying temperature is 55-60°C.

[0015] Preferably, the parameters of the granulation process are set as follows: feed rate of 10-20 kg / min, rotation speed of 8-10 r / min, discharge rate of 10-20 kg / min, pressure of 2-3 kg / cm 2 , the temperature is 20-30℃.

[0016] The invention also provides application of the rodenticide in preventing and controlling rodent damage.

[0017] The present invention also provides application of the rodenticide prepared by the preparation method in preventing and controlling rodent pests.

[0018] Beneficial effects of the present invention:

[0019] The invention adopts ricin to prepare a botanical rodenticide to control domestic rats, which is the first of its kind in China and has significant social benefits, especially environmental benefits.

[0020] The present invention uses ricin to prepare a plant-based rodenticide. Experimental studies have shown that the effective concentration of ricin is within the range of 0.45-1.8%, with 0.9% being the best in terms of mortality and palatability. In laboratory tests, rats reach a peak of mortality on days 5-6, with a mortality rate of 100% on day 9. The feeding coefficient is 0.59, which is higher than the requirement of Class A efficacy standard of not less than 0.3. Field test results show that the rat killing rate reaches 95.17% on day 21.

[0021] The extracted ricinine (effective concentration 10%) had an acute oral LD50 effect on rats 50 The concentration is 68.26-76.96 mg / kg, which is moderately toxic. It has no acute irritation to rabbit skin and is slightly irritating to rabbit eyes. The rodenticide of the present invention is safe to use, store and transport.

[0022] The rodenticide prepared by the invention has a decomposition rate of 4.30% under a heat storage temperature of 54±2° C. for 14 days, which is lower than the requirement of 10%, and has qualified stability. DETAILED DESCRIPTION

[0023] The present invention provides the use of ricinine in preparing rodenticide.

[0024] In the present invention, the effective concentration of ricinine is 0.45-1.80%, preferably 1.125%.

[0025] The present invention also provides a rodenticide comprising the following components in parts by weight:

[0026] 0.45 to 1.80 parts of ricinine, preferably 1.125 parts,

[0027] Base bait 97-99, preferably 98 parts,

[0028] 0.1 to 0.3 parts of stabilizer, preferably 0.2 parts,

[0029] The bittering agent is 0.002 to 0.02 parts, preferably 0.011 parts.

[0030] In the present invention, the base bait is one or more of corn flour, milk powder, peanut cake, and vegetable oil; the stabilizer is butylated hydroxytoluene; and the bittering agent is denatonium benzoate.

[0031] In the present invention, when the base bait is corn flour, milk powder, peanut cake and vegetable oil, the mass ratio of corn flour, milk powder, peanut cake and vegetable oil is 40-44:0.5-1.5:2-4:3-5, preferably 42:1:3:4.

[0032] In the present invention, the ricinine extraction process comprises the following steps: taking 500g of castor meal, adding 5L of deionized water, decocting for 3h, filtering, and cooling the filtrate to 25°C to obtain a ricinine extract; passing the ricinine extract through a D101 macroporous adsorption resin column, wherein the macroporous resin column has a volume of 500mL, a diameter of 5cm, and a diameter-to-height ratio of 1:10; after the adsorption is completed, 4BV of the ricinine extract is eluted with water at a flow rate of 3.5BV / h to remove impurities, and then 4BV of the ricinine extract is eluted with 32% ethanol, and the 32% ethanol eluate is collected and the solvent is recovered to 50mL at 67°C and -0.12MPa under reduced pressure to obtain ricinine.

[0033] The present invention also provides a method for preparing the rodenticide, comprising the following steps:

[0034] (1) mixing ricin, base bait, stabilizer, and bittering agent to obtain a mixture;

[0035] (2) granulating the mixture and drying it to obtain a rodenticide.

[0036] In the present invention, the drying method is heating in a dryer; the drying temperature is 55-60°C, preferably 57.5°C.

[0037] In the present invention, the parameters of the granulation process are set as follows: feed rate of 10-20 kg / min, preferably 15 kg / min, rotation speed of 8-10 r / min, preferably 9 r / min, discharge rate of 10-20 kg / min, preferably 15 kg / min, pressure of 2-3 kg / cm 2 , preferably 2.5kg / cm 2 , the temperature is 20-30℃, preferably 25℃.

[0038] The invention also provides application of the rodenticide in preventing and controlling rodent damage.

[0039] The present invention also provides application of the rodenticide prepared by the preparation method in preventing and controlling rodent pests.

[0040] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0041] Example 1 A rodenticide

[0042] Extraction of ricinine: 500g of castor meal is added to 5L of deionized water, boiled for 3h, filtered, and the filtrate is cooled to 25°C to obtain the ricinine extract; the ricinine extract is passed through a D101 macroporous adsorption resin column with a volume of 500mL, a diameter of 5cm, and a diameter-to-height ratio of 1:10. After adsorption is completed, 4BV of impurities are removed by elution with water at a flow rate of 3.5BV / h, and then 4BV of impurities are eluted with 32% ethanol. The 32% ethanol eluate is collected and the solvent is recovered to 50mL under reduced pressure at 67°C and -0.12MPa to obtain ricinine.

[0043] Preparation method of rodenticide:

[0044] (1) 0.45 kg of ricinine, 86 kg of corn flour, 8 kg of vegetable oil, 1 kg of milk powder, 4 kg of peanut cake, 0.30 kg of butylated hydroxytoluene, and 0.02 kg of denatonium benzoate were mixed to obtain a mixture;

[0045] (2) The mixture was fed at a rate of 13 kg / min, a rotation speed of 8 r / min, a discharge rate of 13 kg / min, and a pressure of 2 kg / cm 2 , granulate at 20°C, heat and dry in a dryer at 60°C to obtain a rodenticide.

[0046] Detected by high-pressure liquid chromatography, the effective concentration of the extracted ricinine is 10%; the effective concentration of ricinine in the rodenticide is 0.45%.

[0047] Example 2 A rodenticide

[0048] The steps of extracting ricin are as follows:

[0049] Preparation method of rodenticide:

[0050] (1) Mix 0.9 kg of ricinine, 82 kg of corn flour, 8 kg of vegetable oil, 2 kg of milk powder, 6 kg of peanut cake, 0.25 kg of butylated hydroxytoluene, and 0.018 kg of denatonium benzoate;

[0051] (2) The mixture was stirred at a feed rate of 15 kg / min, a rotation speed of 9 r / min, a discharge rate of 15 kg / min, and a pressure of 3 kg / cm 2 , granulate at 25°C, heat and dry in a dryer at 55°C to obtain a rodenticide.

[0052] Detected by high pressure liquid chromatography, the effective concentration of ricinine in the rodenticide is 0.90%.

[0053] Example 3 A rodenticide

[0054] The steps of extracting ricin are as follows:

[0055] Preparation method of rodenticide:

[0056] (1) Mix 1.8 kg of ricinine, 82 kg of corn flour, 6 kg of vegetable oil, 3 kg of milk powder, 8 kg of peanut cake, 0.20 kg of butylated hydroxytoluene, and 0.01 kg of denatonium benzoate;

[0057] (2) The mixture was stirred at a feed rate of 17 kg / min, a rotation speed of 10 r / min, a discharge rate of 17 kg / min, and a pressure of 2.5 kg / cm 2 , granulate at 30°C, heat and dry in a dryer at 57°C to obtain a rodenticide.

[0058] Detection by high pressure liquid chromatography showed that the effective concentration of ricinine in the rodenticide was 1.80%.

[0059] Experimental Example 1: Rodenticide Test

[0060] 1. Laboratory Test

[0061] 1.1 Experimental mice: 40 SD rats, half male and half female (purchased from the Experimental Animal Center of Lanzhou Medical University, weighing 100-120 g).

[0062] 1.2 Test Methods: Referring to "Test Methods and Evaluation of the Efficacy of Registered Pesticide Rodenticides for Control of Housed Rodents," NY / T 1152-2006, four cages were set up, namely, Example 1, Example 2, Example 3, and a control group. Ten normal SD rats, half male and half female, were randomly placed in each cage. 50 g of the rodenticide described in Examples 1-3 and 50 g of rat feed were placed symmetrically in each cage. Bait consumption and replenishment were recorded daily, and the positions of the bait boxes were swapped. Observation was continued for 7 days. After the rodenticide was removed, normal feeding was resumed for 14 days. Daily mortality was observed and recorded, and the food intake coefficient and mortality rate were calculated. A control group was set up, which only contained feed, with all other aspects being the same as above.

[0063] Calculation formula:

[0064] Mouse density = number of positive powder blocks / number of effective powder blocks,

[0065] Mortality rate = (rat density before the test - rat density after the test) / rat density before the test,

[0066] Feeding coefficient = rodenticide consumption / feed consumption, grading standard: Grade A: feeding coefficient > 0.3, Grade B: 0.1 ≤ feeding coefficient ≤ 0.3;

[0067] Adjusted mortality rate = (mortality rate of the embodiment - mortality rate of the control group) / (1 - mortality rate of the control group), grading standard: Grade A: adjusted mortality rate > 90%, Grade B: 80% ≤ adjusted mortality rate ≤ 90%.

[0068] 1.3 Experimental results: The calculation results of indoor efficacy determination are shown in Table 1.

[0069] Table 1 Results of indoor efficacy tests on ricinine rodenticides at different concentrations

[0070] As can be seen from Table 1, in Examples 1 to 3, the rat mortality peaked on days 5 to 8, and the mortality rate reached over 90% on days 9 to 12, indicating a significant rodenticide effect. The feeding coefficients were all above 0.52, exceeding the standard feeding coefficient requirement of 0.30 for Grade A, indicating that the rodenticide had excellent palatability.

[0071] When the effective concentration of ricinine was 0.9%, the rodenticide had the most significant effect, with a feeding coefficient of 0.59 and a mortality rate of 100% on the 9th day.

[0072] 2. Outdoor test

[0073] 2.1 Experimental location: Luzhou pig farm, Sichuan, with a total of 110 pig houses. Nine pig houses with a higher visual rat density were selected, and three of them were randomly selected to represent the experimental area (the bait was the rodenticide described in Example 1), the control area (the bait was 0.005% bromadiolone wax block rodenticide, purchased from Guangxi Longhua Xinye Sanitary Insecticide Co., Ltd., batch number 20210912), and the blank control area (the same amount of pig feed was fed according to the feeding method).

[0074] 2.2 Test method: Powder method is calculated based on 300 pieces, and the positive rate is ≥20%. Before the experiment, the powder method is used to investigate the rat density of dominant rat species (positive powder pieces / effective powder pieces). Test area, indoor every 15m 2 Place three piles of the corresponding bait (as described in 2.1) outdoors. Place one pile of the rodenticide described in Example 1 every 8 m, 15 g per pile, in a poison bait box. Place the bait for 5 consecutive days and check the bait consumption and replenish it. The control area and the blank control area are placed in the same manner as above. 21 days after the baiting, investigate the rat density in the test area and the control area, and use x 2 Statistical analysis was performed using the test pair. P>0.05 indicated no difference.

[0075] 2.3 Test results: The results of on-site efficacy test are shown in Table 2.

[0076] Table 2 Results of on-site comparative efficacy test of ricinine rodenticides

[0077] As can be seen from Table 2, both the rodenticide described in Example 1 and the control agent have good rodenticide effects, with a corrected rodenticide rate of 95.17%, which is consistent with the rodenticide rate of the control drug bromadiolone (no statistical difference).

[0078] Experimental Example 2 Stability Test

[0079] The relevant methods in "Determination of Thermal Storage Stability of Pesticides" GB / T 19136-2021 were followed. 20 g of the rodenticides described in Examples 1 to 3 (corresponding to Samples 1 to 3, respectively) were sealed in 25 mL test tubes and placed in a thermostat set at 54°C ± 2°C. After 14 days of thermal storage, the active ingredient content of the samples after thermal storage was tested and compared with the active ingredient content before thermal storage. The thermal storage decomposition rate was calculated as: thermal storage decomposition rate = (pre-storage content - post-storage content) / pre-storage content. The results are shown in Table 3.

[0080] Table 3 Results of heat storage stability test of ricinine rodenticide samples

[0081] The results showed that the product had an average decomposition rate of 4.30% after 14 days of heat storage at 54±2℃, which was lower than the requirement of 10%, and the stability was qualified.

[0082] Experimental Example 3 Toxicology Test

[0083] Testing basis: GB15670-1995 Toxicological test methods for pesticide registration.

[0084] 1. Acute oral toxicity test

[0085] 1.1 Experimental animals: SD rats (purchased from the Experimental Animal Center of Lanzhou Medical University, ordinary grade), weighing 100 g to 120 g.

[0086] 1.2 Test samples: Take ricinine (effective concentration 10%) obtained by the extraction method described in Example 1, dilute and mix the ricinine with distilled water, and divide it into four concentration groups of 25 mg / kg, 50 mg / kg, 100 mg / kg, and 200 mg / kg according to 2.0 geometric progressions.

[0087] 1.3 Test method: Take rats that have been raised (temperature 25°C, humidity 55%) for one week and have no abnormalities. Fast for 12 hours before exposure, measure body weight, and then randomly divide them into 4 groups, 10 rats in each group (half male and half female). Orally gavage once at 1.0 mL / 100 g body weight, and the control group gavage distilled water. After administration, observe for symptoms of poisoning such as tremors, convulsions, and difficulty breathing. Observe for 2 weeks and record the number of deaths. Calculate LD according to the modified Coulter method. 50 .

[0088] 1.4 Test results: The results of the acute oral toxicity test are shown in Table 4.

[0089] Table 4 Results of acute oral toxicity test of 10% ricinine mother solution

[0090] The results showed that approximately 2 hours after administration, some rats in the high-concentration group developed symptoms of poisoning, including lethargy and ruffled fur. Symptoms became more pronounced 24 to 72 hours after exposure, including agitation, paroxysmal breathing difficulties, and convulsions. These symptoms were followed by lethargy, curling up, and death. Rats in the control group showed no symptoms of poisoning.

[0091] The acute oral LD50 of ricinine (effective concentration 10%) extracted by the extraction method of Example 1 was 50 : Females: 76.96 mg / kg; males: 68.26 mg / kg. According to my country's pesticide toxicity classification standards, it is moderately toxic. This indicates that it is safe to use as a raw material for pharmaceutical preparations.

[0092] 2. Acute skin irritation test

[0093] 2.1 Experimental animals: New Zealand white rabbits (purchased from the Experimental Animal Center of Xijing Hospital), weighing 2 kg to 3 kg.

[0094] 2.2 Test method:

[0095] 1) Four healthy rabbits were selected and raised in an experimental environment (temperature 25°C, humidity 55%) for 24 hours before the experiment.

[0096] 2) Cut the hair on both sides of the spine of each rabbit into 2×6cm 2 ;

[0097] 3) After 24 hours, 0.5 mL of ricinine (effective concentration 10%) obtained by the extraction method described in Example 1 was applied to the hairless skin on one side of the spine of each rabbit. The skin was then covered with gauze and secured with adhesive tape. The other side of the skin was coated with distilled water as a control.

[0098] 4) Rinse with warm water 4 hours after application and gently wipe with gauze to remove any residual drug on the skin;

[0099] 5) Observe and record the formation of erythema and edema on the test skin 1 hour, 24 hours, and 72 hours after ricinine removal, and the recovery of erythema and edema on the test skin on the 7th and 14th days.

[0100] 2.3 Test results: The results of the acute skin irritation test are shown in Table 5.

[0101] Table 5 Results of acute skin irritation test of 10% ricinine mother solution

[0102] Erythema formation: No erythema was observed on the test skin 1 to 72 hours after ricinine removal. Edema formation: No edema was observed on the test skin 1 to 72 hours after ricinine removal. No abnormalities were observed on the skin of the control group. This indicates that the drug is safe for use as a raw material in pharmaceutical preparations.

[0103] 3. Eye irritation test

[0104] 3.1 Experimental animals: New Zealand white rabbits (purchased from the Experimental Animal Center of Xijing Hospital), weighing 2 kg to 3 kg.

[0105] 3.2 Test method:

[0106] 1) Four healthy rabbits were selected and raised in an experimental environment (temperature 25°C, humidity 55%) for 24 hours before the experiment.

[0107] 2) Gently pull down one eyelid of each rabbit, take 0.1 mL of ricinine (effective concentration 10%) obtained by the extraction method described in Example 1 and drip it into the conjunctival sac. Immediately, gently close the eyelid for 1 minute. The other eye is treated with distilled water as a control.

[0108] 3) Observe the degree of irritation reaction of the cornea, iris, and conjunctiva at 1 hour, 24 hours, 48 ​​hours, and 72 hours after administration, and observe the recovery of the reaction on the 4th and 7th days;

[0109] 4) Do not wash eyes within 24 hours after administration.

[0110] 3.3 Test results: The results of the eye irritation test are shown in Table 6.

[0111] Table 6 Results of eye irritation test on 10% ricinine mother solution

[0112] One to 12 hours after application, the conjunctivae of the test animals showed bright red congestion, mild edema, and a small amount of secretion; the irises and corneas were normal. Conjunctival congestion was still visible in some animals 24 hours after application, but this had largely returned to normal by 48 hours. The eyes of the control group were normal. This indicates that the drug is safe for use as a raw material in pharmaceutical preparations.

[0113] As can be seen from the above examples, the present invention provides the application of ricin, a rodenticide, and a preparation method and application. The rodenticide prepared by the present invention has a feeding coefficient of 0.52 or above, which is higher than the requirement of the Class A efficacy standard of not less than 0.3. The results of field tests show that the rodenticide rate reaches 95.17% in 21 days. Moreover, the rodenticide of the present invention is safe to use, store and transport, and has qualified stability.

[0114] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. Application of ricinine in the preparation of rodenticides.

2. The use according to claim 1, wherein the effective concentration of ricinine is 0.45-1.80%.

3. A rodenticide, characterized in that: The invention comprises the following components in parts by mass: 0.45 to 1.80 parts of ricinine, 97 to 99 parts of base bait, 0.1 to 0.3 parts of stabilizer and 0.002 to 0.02 parts of bittering agent.

4. The rodenticide according to claim 3, characterized in that The base bait is one or more of corn flour, milk powder, peanut cake, and vegetable oil; the stabilizer is butylated hydroxytoluene; and the bittering agent is denatonium benzoate.

5. The rodenticide according to claim 3 or 4, characterized in that The effective concentration of ricinine in the rodenticide is 0.45-1.80%, preferably 0.90%.

6. The method for preparing the rodenticide according to any one of claims 3 to 5, characterized in that: The steps include: (1) mixing ricin, base bait, stabilizer, and bittering agent to obtain a mixture; (2) granulating the mixture and drying it to obtain a rodenticide.

7. The preparation method according to claim 6, characterized in that The drying method is heating in a dryer; the drying temperature is 55-60°C.

8. The preparation method according to claim 7, characterized in that The parameters of the granulation process are set as follows: feed rate of 10-20 kg / min, rotation speed of 8-10 r / min, discharge rate of 10-20 kg / min, pressure of 2-3 kg / cm 2 , the temperature is 20-30℃.

9. Use of the rodenticide according to any one of claims 3 to 5 in preventing and controlling rodent pests.

10. Use of the rodenticide prepared by the preparation method according to any one of claims 6 to 8 in preventing and controlling rodent pests.

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