Water-soluble diclazuril solution and preparation method thereof

Abstract

The invention discloses a water-soluble diclazuril solution and a preparation method thereof, and belongs to the technical field of veterinary drug preparations, the water-soluble diclazuril solution comprises 10-30 g / L of diclazuril, 100-500 g / L of an organic solvent, 30-60 g / L of an amine organic base, 5-10 g / L of a surfactant, the balance of water for injection, 3-5 g / L of propyl gallate, and 10-15 g / L of a composite fermentation broth, the organic solvent comprises the following components in percentage by mass: 20-40% of N, N-dimethylformamide, 40-70% of propylene glycol and 10-20% of ethanol, and the compound fermentation liquor is a fermentation product obtained by fermenting a mixture of felwort and tamarind fruits with a compound microbial inoculant. The water-soluble diclazuril solution provided by the invention not only can effectively prevent and treat chicken coccidiosis, but also is beneficial to improving the weight gain rate of broiler chickens in the recovery process after infection, the preparation method is simple, the cost is low, the use mode is convenient and efficient, and the use value of the water-soluble diclazuril solution in the aspect of broiler chicken breeding is effectively improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of veterinary preparations, in particular to a water-soluble diclazuril solution and a preparation method thereof. BACKGROUND

[0002] Chicken coccidiosis is a common parasitic disease caused by Eimeria parasitizing in the intestinal tract of chickens, which is one of the common diseases affecting the economic benefits of the poultry industry. Diclazuril is a highly effective and special anticoccidial drug for preventing and treating chicken coccidiosis, and belongs to a highly effective and low-toxicity triazine anticoccidial drug.

[0003] The following obstacles and defects exist in the preparation and use of a drug with diclazuril as the core: on the one hand, diclazuril is extremely difficult to dissolve in water (solubility <0.1 mg / L), in order to improve the solubility of diclazuril in the drug, the traditional preparation method needs to use heating means, and the existing technology is improved through solid dispersion technology and nano-suspension technology, and there are problems of complex process and high manufacturing cost; on the other hand, chicken coccidiosis not only directly damages the intestinal tissue of chickens, leading to death, but also causes bacterial flora disorder in the intestinal tract of chickens during the infection period, making it difficult for intestinal probiotics to colonize and survive, and diclazuril is a broad-spectrum anticoccidial drug, which, while exerting anticoccidial effect, also inhibits or even kills part of the intestinal probiotics, and the double factors together cause the broilers to fail to complete effective feed conversion during the infection period, thereby affecting the breeding income. In view of the above problems, the present application provides a water-soluble diclazuril solution and a preparation method thereof. SUMMARY

[0004] The purpose of the present application is to provide a water-soluble diclazuril solution and a preparation method thereof in order to solve the above problems.

[0005] The present application achieves the above-mentioned purpose through the following technical solutions: The present application provides a water-soluble diclazuril solution, the composition of the water-soluble diclazuril solution comprising: 10-30 g / L of diclazuril, 100-500 g / L of an organic solvent, 30-60 g / L of an amine organic base, 5-10 g / L of a surfactant, and the balance being water for injection. The composition of the organic solvent comprises, by mass percentage: 20-40% of N,N-dimethylformamide, 40-70% of propylene glycol, and 10-20% of ethanol.

[0006] As a further optimization scheme of the present application, the surfactant is polyoxyethylene castor oil or Tween 80.

[0007] As a further optimization scheme of the present application, the amine organic base is diethanolamine.

[0008] As a further optimization scheme of the present application, the composition of the water-soluble diclazuril solution further comprises propyl gallate 3-5 g / L.

[0009] As a further optimization scheme of the present application, the composition of the water-soluble diclazuril solution further comprises compound fermentation liquor 10-15 g / L, the compound fermentation liquor being a fermentation product obtained by fermenting a mixture of gentian and tamarind with a compound microbial agent, the compound microbial agent being a mixture of Saccharomyces boulardii, Butyribacterium, and Lactobacillus rhamnosus in a ratio of 1:1:1.

[0010] The present application further provides a preparation method of the water-soluble diclazuril solution. (1) mixing N,N-dimethylformamide, propylene glycol, and ethanol to obtain an organic solvent; (2) adding diclazuril into the organic solvent, stirring until dissolved to obtain solution A; (3) adjusting the pH value of solution A to 8-9.5 with an amine organic base, stirring uniformly, then adding surfactant and again stirring thoroughly, then adding water for injection to constant volume, mixing uniformly to obtain a mixed solution; (4) filtering and filling the mixed solution in sequence to obtain the water-soluble diclazuril solution.

[0011] As a further optimization scheme of the present application, step (2) further comprises uniformly mixing propyl gallate into the diclazuril after dissolution.

[0012] As a further optimization scheme of the present application, step (1) further comprises preparing compound fermentation liquor, uniformly mixing the compound fermentation liquor into the organic solvent to obtain a mixed solvent, which is used to replace the organic solvent in step (2).

[0013] As a further optimization scheme of the present application, the preparation method of the compound fermentation liquor is as follows: mixing gentian extract and tamarind paste in a mass ratio of 2:1, sterilizing the mixture to obtain a fermentation substrate, mixing Saccharomyces boulardii, Butyribacterium, and Lactobacillus rhamnosus in a ratio of 1:1:1 to obtain a mixed microbial agent, inoculating the mixed microbial agent into the fermentation substrate at an inoculation amount of 8-10%, and fermenting under anaerobic conditions at 33-35℃ for 24-36 h, then filtering to obtain the fermentation product.

[0014] The present application has the following beneficial effects: (1) The water-soluble diclazuril solution and the preparation method thereof, by using the organic solvent mixed by N, N-dimethylformamide, propylene glycol and ethanol, the solubility of diclazuril in the water-soluble diclazuril solution is effectively improved, and the low irritation is considered, so that the water-soluble diclazuril solution can not only effectively prevent and treat chicken coccidiosis, but also is beneficial to improve the weight gain rate of broilers in the recovery process after infection, and the preparation process of the water-soluble diclazuril solution does not need heating, the operation is simple, the production cost is low, and the water-soluble diclazuril solution can be directly used for oral administration or mixed drinking administration, which is convenient to use. (2) By mixing the mixture of gentian extract and tamarind mud as the fermentation substrate into the water-soluble diclazuril solution, the use effect of the water-soluble diclazuril solution is further improved, which is specifically that the weight gain rate of broilers in the recovery process after administration is higher, the use value of the water-soluble diclazuril solution in broiler breeding is effectively improved, and the influence of chicken coccidiosis on breeding income is reduced. DETAILED DESCRIPTION

[0015] It is necessary to point out here that the following detailed description is only used to further illustrate the application, and cannot be understood as limiting the protection scope of the application, and the skilled in the art can make some non-essential improvements and adjustments to the application according to the above application content.

[0016] I. Experimental materials (1) Preparation of gentian extract: dry gentian grass into powder, add purified water according to the solid-liquid ratio of 1:20 (g / mL), and extract twice at 95 DEG C for 15 min each time, filter the extract to obtain gentian extract.

[0017] (2) Preparation of tamarind mud: shell and tendon membrane of fresh tamarind are removed, and then crushed and ground to obtain tamarind mud.

[0018] (3) The mature commercial products of the three strains of bacteria, i.e. Saccharomyces boulardii, Butyribacterium and Lactobacillus rhamnosus, have their own activation conditions: YPD medium is used for Saccharomyces boulardii, and the activation is carried out at 30 DEG C for 6 hours; MRS medium is used for Butyribacterium and Lactobacillus rhamnosus, and the activation is carried out under anaerobic conditions at 37 DEG C for 8 hours; after activation, the bacteria are collected by centrifugation, washed with physiological saline, and then prepared into a bacterial suspension with a concentration of 10 8 CFU / mL.

[0019] The method used in the application is a conventional method known to those skilled in the art, and the reagents and materials used are commercially available products unless otherwise specified.

[0020] II. Experimental method 1. A water-soluble diclazuril solution Example 1 A water-soluble diclazuril solution, the preparation method thereof specifically comprises the following steps: (1) uniformly mix 20% of N,N-dimethylformamide, 70% of propylene glycol and 10% of ethanol according to the mass percentage, to obtain an organic solvent; (2) add 2g of diclazuril into 30g of the organic solvent, stir until dissolved, to obtain solution A; (3) adjust the pH value of solution A to 9 by using diethanolamine (the amount is about 4.5g), after uniform stirring, continue to add 0.8g of polyoxyethylene castor oil, after fully stirring again, supplement with water for injection to 100ml, after uniform mixing, obtain a mixed solution, and after the mixed solution is sequentially subjected to filtration and filling treatment, a water-soluble diclazuril solution is obtained.

[0021] The above preparation steps are all carried out under light-proof and sterile conditions, the same below.

[0022] Example 2 A water-soluble diclazuril solution, the preparation method thereof specifically comprises the following steps: (1) uniformly mix 30% of N,N-dimethylformamide, 50% of propylene glycol and 20% of ethanol according to the mass percentage, to obtain an organic solvent; (2) add 1g of diclazuril into 10g of the organic solvent, stir until dissolved, to obtain solution A; (3) adjust the pH value of solution A to 8 by using diethanolamine (the amount is about 3g), after uniform stirring, continue to add 0.5g of polyoxyethylene castor oil, after fully stirring again, supplement with water for injection to 100ml, after uniform mixing, obtain a mixed solution, and after the mixed solution is sequentially subjected to filtration and filling treatment, a water-soluble diclazuril solution is obtained.

[0023] Example 3 A water-soluble diclazuril solution, the preparation method thereof specifically comprises the following steps: (1) uniformly mix 40% of N,N-dimethylformamide, 40% of propylene glycol and 20% of ethanol according to the mass percentage, to obtain an organic solvent; (2) add 3g of diclazuril into 50g of the organic solvent, stir until dissolved, to obtain solution A; (3) adjust the pH value of solution A to 9.5 by using diethanolamine (the amount is about 6g), after uniform stirring, continue to add 1g of polyoxyethylene castor oil, after fully stirring again, supplement with water for injection to 100ml, after uniform mixing, obtain a mixed solution, and after the mixed solution is sequentially subjected to filtration and filling treatment, a water-soluble diclazuril solution is obtained.

[0024] 2. Efficacy test of water-soluble diclazuril solution (1) Test subjects Ten-day-old healthy Hy-Line laying hens (purchased from Zhanjiang Jinsheng Animal Husbandry Technology Co., Ltd.) were used as test subjects, and were fed for 20 days. During this period, the feed was a complete feed for chicks without any antibacterial drugs, and the hens were allowed to drink and eat freely. After 20 days, the test began, and no vaccines, vaccines or drugs were used during the test.

[0025] (2) Test process The following experimental groups were set up: ① Blank group (no inoculation of strains and no use of drugs); ② Positive group (inoculation of strains and no use of drugs); ③ Drug administration group A (inoculation of strains and administration of the drug prepared in Example 1); ④ Drug administration group B (inoculation of strains and administration of the drug prepared in Example 2); ⑤ Drug administration group C (inoculation of strains and administration of the drug prepared in Example 3); Fifty Hy-Line laying hens were assigned to each group as test subjects. During the test, the administration method was mixed drinking (drinking water) administration, and the addition amount was 0.8 ml per hen, which was drunk in a concentrated manner twice a day for 5 days. After one week of administration, the hens were weighed once, then orally inoculated with Eimeria tenella oocysts, and continued to be fed for 15 days after inoculation. Fifteen days later, the surviving hens were weighed again.

[0026] (3) Test results The following indicators were detected and statistically analyzed: ① Mortality rate: During the test, the hens showed typical symptoms of chicken coccidiosis and died, and the carcasses showed typical characteristic lesions. The number of deaths was determined, and the mortality rate was calculated based on the number of deaths in each group. ② Effective rate: During the test, only mild clinical symptoms were observed, and the hens recovered significantly after 2-3 days. The hens ate and drank normally, and their feces were basically normal, which was considered to be a treatment success. The effective rate was calculated accordingly. ③ Weight gain rate: The weight gain rate of each surviving Hy-Line hen was calculated based on the first and second weight data. The average relative weight gain rate of each test hen was calculated based on the blank group.

[0027] The statistical results are shown in the following table. As can be seen from the table, the water-soluble diclazuril solutions obtained in Examples 1-3 can significantly reduce the mortality rate of Hy-Line hens infected with chicken coccidiosis, and can effectively improve the weight gain rate of Hy-Line hens during the recovery period from infection. The reason is inferred as follows: the organic solvent obtained by compounding N,N-dimethylformamide, propylene glycol and ethanol effectively improves the solubility of diclazuril, realizes the effect of giving consideration to low irritation and high solubility, comprehensively optimizes the in-vivo absorption effect of the water-soluble diclazuril solution, and thus realizes better curative effect.

[0028] Group Mortality (%) Efficiency (%) Weight gain rate (%) Blank group / / 100.0 Positive group 60 / 38.9 Dosing group A 4 96 68.3 Dosing group B 6 94 67.7 Dosing group C 4 94 65.5 3. Effect of composition of organic solvent on water-soluble diclazuril solution Based on Example 1, only the composition of the organic solvent therein is adjusted, and Comparative Examples 1-3 are set. Specifically, the composition of the organic solvent in Example 1 and Comparative Examples 1-3 is shown in the following table: Group N,N-dimethylformamide (%) Propylene glycol (%) Ethanol (%) Comparative example 1 - 87.5 12.5 Comparative example 2 67 - 33 Comparative example 3 22 78 - The use effect of the water-soluble diclazuril solution obtained in Comparative Examples 1-3 is detected, and the detection method is the same as in 2. The detection results are shown in the following table. It can be seen from the table that, among the organic solvents, the absence of any one of N,N-dimethylformamide, propylene glycol and ethanol will cause the therapeutic effect of the finally obtained water-soluble diclazuril solution to decrease, and correspondingly, the weight gain rate of the surviving Hyline chickens also decreases. This result shows that the improvement effect of the organic solvent on the efficacy of the water-soluble diclazuril solution is realized based on the compounding of N,N-dimethylformamide, propylene glycol and ethanol.

[0029] Group Mortality (%) Efficiency (%) Weight gain rate (%) Comparative example 1 8 86 56.3 Comparative example 2 14 85 50.4 Comparative example 3 12 86 55.7 4. Effect of compound fermentation liquor on water-soluble diclazuril solution Based on Example 1, Examples 4-6 are set, and the specific settings are as follows: Example 4 A water-soluble diclazuril solution, and a preparation method thereof, specifically includes the following steps: (1) 20% of N,N-dimethylformamide, 70% of propylene glycol and 10% of ethanol are uniformly mixed according to mass percentage to obtain an organic solvent; Gentiana extract and tamarind paste are mixed at a mass ratio of 2:1, and the mixture is subjected to sterilization treatment (pasteurization, 85°C, 5min, and rapid cooling to room temperature after sterilization, the same below) to obtain a fermentation substrate. Activated Saccharomyces boulardii, Butyribacterium and Lactobacillus rhamnosus are mixed at a ratio of 1:1:1 to obtain a mixed inoculum. The mixed inoculum is inoculated into the fermentation substrate at an inoculation amount of 9%, and the fermentation substrate is subjected to fermentation under anaerobic conditions at 34°C for 30h and then subjected to filtration treatment to obtain a fermentation product; 1.3g of the fermentation product is uniformly mixed into 30g of the organic solvent to obtain a mixed solvent; (2) 2g of diclazuril is added to the mixed solvent, stirred until dissolved, and then 4g / L of propyl gallate is uniformly mixed to obtain solution A; (3) adjust the pH value of solution A (about 5.7g in dosage) to 9 with diethanolamine, stir uniformly, then continue to add polyoxyethylene castor oil 0.8g, stir again, then supplement with water for injection to 100ml, mix uniformly to obtain a mixed solution, then filter and fill the mixed solution to obtain a water-soluble diclazuril solution.

[0030] Example 5 A water-soluble diclazuril solution, the preparation method specifically comprising the following steps: (1) uniformly mix 20% N,N-dimethylformamide, 70% propylene glycol and 10% ethanol according to the mass percentage to obtain an organic solvent; Mix the gentian extract and tamarind paste according to a mass ratio of 2:1, sterilize the mixture to obtain a fermentation substrate, mix the activated Saccharomyces boulardii, Butyribacterium and Lactobacillus rhamnosus according to a ratio of 1:1:1 to obtain a mixed bacterial agent, inoculate the mixed bacterial agent into the fermentation substrate according to an inoculation amount of 8%, and then ferment under anaerobic conditions at 33°C for 24-36h, then filter to obtain a fermentation product; Mix 1g of the fermentation product into 30g of the organic solvent to obtain a mixed solvent; (2) add 2g of diclazuril into the mixed solvent, stir until dissolved, then uniformly mix in 3g / L propyl gallate to obtain solution A; (3) adjust the pH value of solution A (about 5.2g in dosage) to 9 with diethanolamine, stir uniformly, then continue to add polyoxyethylene castor oil 0.8g, stir again, then supplement with water for injection to 100ml, mix uniformly to obtain a mixed solution, then filter and fill the mixed solution to obtain a water-soluble diclazuril solution.

[0031] Example 6 A water-soluble diclazuril solution, the preparation method specifically comprising the following steps: (1) uniformly mix 20% N,N-dimethylformamide, 70% propylene glycol and 10% ethanol according to the mass percentage to obtain an organic solvent; Mix the gentian extract and tamarind paste according to a mass ratio of 2:1, sterilize the mixture to obtain a fermentation substrate, mix the activated Saccharomyces boulardii, Butyribacterium and Lactobacillus rhamnosus according to a ratio of 1:1:1 to obtain a mixed bacterial agent, inoculate the mixed bacterial agent into the fermentation substrate according to an inoculation amount of 8-10%, and then ferment under anaerobic conditions at 35°C for 24-36h, then filter to obtain a fermentation product; Mix 1.5g of the fermentation product into 30g of the organic solvent to obtain a mixed solvent; (2) 2g diclazuril was added into the mixed solvent, stirred until dissolved, and then 5g / L propyl gallate was uniformly mixed to obtain solution A; (3) The pH value of solution A (about 6g) was adjusted to 9 using diethanolamine, and after uniform stirring, 0.8g of polyoxyethylene castor oil was added, and after uniform stirring, water for injection was added to 100ml, and after uniform mixing, a mixed solution was obtained. The mixed solution was sequentially filtered and filled to obtain a water-soluble diclazuril solution.

[0032] Positive control group The water-soluble diclazuril solution prepared in Example 1 was administered, and 48 hours after administration, the test subjects were supplemented with probiotics, and the probiotics used were avian compound probiotics from Huachu, and the supplementing method was to add them to the drinking water, and the addition amount was 4g per chicken, and the chickens concentrated on drinking.

[0033] The use effect of the water-soluble diclazuril solution obtained in Examples 4-6 was detected, the method was the same as 2, and the treatment effect of the positive control group was also detected, and the detection method was the same as above. The detection results are shown in the following table, and from the table it can be seen that: Compared with Example 1 (i.e. administration group A) and the positive control group, the water-soluble diclazuril solution obtained in Examples 4-6 has better curative effect, and significantly improves the weight gain rate of Hailan laying hens during the period from infection to recovery, and the water-soluble diclazuril solution has better effect on improving the weight gain rate than the conventional treatment scheme of administering first and then supplementing intestinal probiotics; The reason is that the probiotics and other active ingredients in the compound fermentation liquor can not only supplement part of the probiotics to the chicken intestinal tract, but also optimize the intestinal environment, thereby comprehensively reducing the damage of chicken coccidiosis to the chicken intestinal function, and finally showing the improvement of the weight gain rate.

[0034] Group Mortality (%) Efficiency (%) Weight gain rate (%) Example 4 0 100 97.3 Example 5 2 98 96.2 Example 6 0 98 96.8 Positive control group 4 94 72.7 5, Effect of composition of compound fermentation liquor on efficacy of water-soluble diclazuril solution Based on Example 4, the following comparative examples were set: ① Comparative Example 4: The fermentation substrate was replaced with an equal amount of gentian extract; ② Comparative Example 5: The fermentation substrate was replaced with an equal amount of tamarind mud; ③ Comparative Example 6: The fermentation product was replaced with an equal amount of mixed bacterial liquid, and the mixed bacterial liquid was a mixture of water for injection and mixed bacterial agent (the amount was the same as that of Example 4).

[0035] The use effect of the water-soluble diclazuril solution obtained in the positive control group and Comparative Examples 4-6 is detected, and the detection results are shown in the following table. As can be seen from the table, in the detection results of Comparative Examples 4-6, the mortality and effective rate data are comparable to those of Example 4, but the weight gain rate data are all reduced by different amplitudes, that is, the weight gain rate improvement effect achieved by the gentian extract, tamarind mud and mixed fermentation of the mixed bacterial agent is better than that of the single fermentation substrate scheme and the scheme of only supplementing the mixed bacterial agent. The results show that the improvement effect of the composite fermentation liquid on the efficacy of the water-soluble diclazuril solution is achieved based on the combined reaction of various components and strains in the fermentation system.

[0036] Group Mortality (%) Efficiency (%) Weight gain rate (%) Comparative example 4 2 96 86.6 Comparative example 5 2 96 82.9 Comparative example 6 4 98 74.1 The above-described examples only express several embodiments of the present application, and the description is more specific and detailed, but it cannot be understood as limiting the scope of the patent of the present application. It should be noted that for ordinary skilled persons in the art, several modifications and improvements can be made without departing from the concept of the present application, which are within the protection scope of the present application.

Claims

1. A water-soluble dicrizoline solution, characterized in that: The water-soluble diclazuril solution comprises: diclazuril 10-30 g / L, organic solvent 100-500 g / L, amine organic base 30-60 g / L, surfactant 5-10 g / L, and the balance being water for injection; The organic solvent comprises, by mass percentage: 20-40% N,N-dimethylformamide, 40-70% propylene glycol, and 10-20% ethanol.

2. The water-soluble diclazuril solution according to claim 1, characterized in that: The surfactant is polyoxyethylene castor oil or Tween 80.

3. The water-soluble diclazuril solution according to claim 1, characterized in that: The amine organic base is diethanolamine.

4. The water-soluble diclazuril solution according to claim 1, characterized in that: The water-soluble diclazuril solution also includes 3-5 g / L of propyl gallate.

5. The water-soluble diclazuril solution according to claim 1, characterized in that: The water-soluble diclazuril solution also includes 10-15 g / L of a compound fermentation broth, which is a fermentation product obtained by fermenting a mixture of gentian and tamarind with a compound microbial agent. The compound microbial agent is composed of Saccharomyces boulardii, Butyric acid bacteria, and Lactobacillus rhamnosus in a 1:1:1 ratio.

6. A method for preparing a water-soluble diclazuril solution as described in any one of claims 1-5, characterized in that: Specifically, the following steps are included: (1) Mix N,N-dimethylformamide, propylene glycol, and ethanol to obtain an organic solvent; (2) Add diclazuril to an organic solvent and stir until dissolved to obtain solution A; (3) Adjust the pH of solution A to 8-9.5 using amine organic bases, stir evenly, add surfactant and stir thoroughly again, add water for injection, make up to volume, mix evenly to obtain a mixed solution; (4) The mixed solution is filtered and filled in sequence to obtain a water-soluble diclazuril solution.

7. The preparation method according to claim 6, characterized in that: Step (2) further includes uniformly mixing propyl gallate into the solution of diclazuril.

8. The preparation method according to claim 7, characterized in that: Step (1) further includes preparing a composite fermentation broth, uniformly mixing the composite fermentation broth into an organic solvent to obtain a mixed solvent, which is used to replace the organic solvent in step (2).

9. The preparation method according to claim 8, characterized in that: The preparation method of the compound fermentation broth is as follows: Gentian extract and tamarind mud are mixed at a mass ratio of 2:

1. After sterilization, the mixture is used to obtain the fermentation substrate. Saccharomyces boulardii, Butyric acid bacteria, and Lactobacillus rhamnosus are mixed at a ratio of 1:1:1 to obtain a mixed bacterial agent. The mixed bacterial agent is inoculated into the fermentation substrate at an inoculation rate of 8-10%. The mixture is fermented under anaerobic conditions at 33-35℃ for 24-36 hours and then filtered to obtain the fermentation broth, which is the fermentation product.