A freezing-proof type kasugamycin aqueous agent with improved reconstitution rate

Abstract

The present application is to solve the problem of freezing and thawing of spring mycin, and the denatured starch is used as an additive, which is added into the spring mycin fermentation extract, and the excellent resolubility is shown after freezing and thawing, and the problem of freezing and precipitation of spring mycin aqueous solution in winter storage in northern China is solved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of modified treatment of agricultural fungicides, in particular to the addition modification of kasugamycin and the preparation method of modified kasugamycin aqueous preparation. BACKGROUND

[0002] As a green, efficient, low-toxicity, low-residue agricultural antibiotic fungicide, the product of kasugamycin belongs to a biological pesticide product prepared by microbial fermentation. At present, the common aqueous formulations on the market are 2% and 6% kasugamycin aqueous products. The demand for 6% kasugamycin is relatively large, and it has a good market prospect. Due to the low temperature in winter in northern China, kasugamycin aqueous agent will freeze and have a precipitate during winter storage. Therefore, without the addition of antifreeze components, the product will cause degradation of drug efficacy and low reconstitution rate, and cause high storage cost. In the prior art, substances such as ethylene glycol, propylene glycol, glycerol, urea, and polyvinyl alcohol are often used as antifreeze agents. For example, the antifreeze agents disclosed in patent documents CN102972402A and CN101248794A use polyvinyl alcohol, ethylene glycol, propylene glycol, and glycerol. In the technical field of pesticide fungicides and their compositions, the above-mentioned antifreeze agents are often used, and the development and research of antifreeze agents in the field of pesticide fungicides and their compositions are rarely focused on research and development. Although the traditional antifreeze agents such as polyvinyl alcohol, ethylene glycol, propylene glycol, and glycerol used in the prior art have good antifreeze effect, there are still some precipitates under low-temperature freezing conditions, and the precipitates need to be shaken or stirred for a long time after thawing to achieve complete reconstitution. Therefore, the existing antifreeze components still have room for improvement in terms of the antifreeze effect of kasugamycin. SUMMARY

[0003] The present application adds polysaccharide auxiliary denatured starch during product extraction to achieve the effects of antifreeze and improved reconstitution rate. By using polysaccharide auxiliary to change the solubility and dissolution rate of kasugamycin, the stability of kasugamycin aqueous product is increased, the storage of kasugamycin aqueous product in winter is more convenient and the cost is reduced, and the drug efficacy is not degraded when the product is used after the temperature warms up.

[0004] The present application provides a kasugamycin aqueous preparation capable of improving the reconstitution rate, and specifically, the kasugamycin fermentation liquor is prepared into filtrate ① by pretreatment of the fermentation liquor and plate and frame filtration during the extraction process, and then the filtrate ① is connected to a storage tank to obtain kasugamycin fermentation stock solution, and then different potency resolving liquids ② are prepared by ion exchange process, the resolving liquids ② are connected to different storage tanks according to the different potencies, when the temperature of the resolving liquid ② reaches about 40 DEG C, 4%-6% of polysaccharide auxiliary denatured starch is added into the storage tank according to the potency of the resolving liquid ②, and then the mixture is uniformly mixed by using an electric mixer to obtain material liquid ③, the potency and the pH value of the material liquid ③ are determined, the pH value is controlled in the range of 3-5, and then the material liquid ③ is purified and concentrated by membrane filtration to obtain kasugamycin anti-freezing aqueous preparation products containing 2% and 6% of kasugamycin and polysaccharide auxiliary denatured starch. The 4%-6% of polysaccharide auxiliary denatured starch is that the mass ratio of denatured starch to kasugamycin fermentation extraction liquid is 4%-6%. The denatured starch is cationic starch, and the cationic starch is prepared by the following steps: corn starch is added into water and stirred and dispersed, enzymes are added for enzymolysis, the enzymolysis time is 20-30 minutes to obtain enzymolysis starch liquid; the enzymolysis starch liquid is heated to 40-50 DEG C, then 6-8% of alkali liquor is added, the temperature is increased to 70-90 DEG C, 2,3-epoxypropane trimethylammonium chloride is added and reacts for 2-4 hours, and finally the cationic modified starch is obtained after acid neutralization.

[0005] The present application has the following beneficial effects:

[0006] The present application adds a polysaccharide auxiliary denatured starch in the product extraction process stage, which can delay the freezing time of the kasugamycin aqueous preparation product, and solve the problems that the precipitate is difficult to be reconstituted in a short time after the temperature is warmed and the original efficacy of the product is maintained, the reconstitution rate is increased from 90% to 100%, and the anti-freezing effect reaches the phenomenon of no freezing within 24 hours under-10 DEG C. BRIEF DESCRIPTION OF DRAWINGS

[0007] Figure 1 It is a kasugamycin aqueous preparation product extraction production process flow chart.

[0008] Figure 2 It is a liquid chromatogram of the kasugamycin product detected by using the liquid phase method.

[0009] Figure 3 is a comparison chart before and after freezing and a reconstitution effect comparison chart of No. 1 and No. 2.

[0010] Figure 4 It is a liquid chromatogram of No. 1 and No. 2 after thawing. DETAILED DESCRIPTION

[0011] Example 1: In the extraction process of kasugamycin fermentation broth, the filtrate ① prepared after the fermentation broth pretreatment and plate and frame filtration processes is connected to a storage tank, and different potency elution solutions ② are prepared through ion exchange processes. The elution solutions ② are connected to different storage tanks according to their potencies. When the temperature of the elution solution ② reaches about 40℃, 4% of the polysaccharide aid, cationic starch, is added to the storage tank according to the potency of the elution solution ②. The mixture is uniformly mixed by using an electric mixer to prepare the feed solution ③. The potency and pH value of the feed solution ③ are determined, and the pH value is controlled in the range of 3-5. Then, the 6% kasugamycin aqueous product is prepared by purifying and concentrating the feed solution ③ through membrane filtration.

[0012] Example 2: In the extraction process of kasugamycin fermentation broth, the filtrate ① prepared after the fermentation broth pretreatment and plate and frame filtration processes is connected to a storage tank, and different potency elution solutions ② are prepared through ion exchange processes. The elution solutions ② are connected to different storage tanks according to their potencies. When the temperature of the elution solution ② reaches about 40℃, 6% of the polysaccharide aid, cationic starch, is added to the storage tank according to the potency of the elution solution ②. The mixture is uniformly mixed by using an electric mixer to prepare the feed solution ③. The potency and pH value of the feed solution ③ are determined, and the pH value is controlled in the range of 3-5. Then, the 6% kasugamycin aqueous product is prepared by purifying and concentrating the feed solution ③ through membrane filtration.

[0013] Example 3: In the extraction process of kasugamycin fermentation broth, the filtrate ① prepared after the fermentation broth pretreatment and plate and frame filtration processes is connected to a storage tank, and different potency elution solutions ② are prepared through ion exchange processes. The elution solutions ② are connected to different storage tanks according to their potencies. When the temperature of the elution solution ② reaches about 40℃, 5% of the polysaccharide aid, cationic starch, is added to the storage tank according to the potency of the elution solution ②. The mixture is uniformly mixed by using an electric mixer to prepare the feed solution ③. The potency and pH value of the feed solution ③ are determined, and the pH value is controlled in the range of 3-5. Then, the 6% kasugamycin aqueous product is prepared by purifying and concentrating the feed solution ③ through membrane filtration.

[0014] The cationic starch in the examples 1-3 is prepared by the following method: 100g of corn starch is added to 500g of water and stirred and dispersed, and enzymes are added for enzymatic hydrolysis. The enzymatic hydrolysis time is 20-30 minutes to obtain an enzymatic hydrolysis starch solution. The enzymatic hydrolysis starch solution is heated to 40-50℃, and then an alkali solution with a mass percentage of 6-8% is added. The temperature is raised to 70-90℃, and 2,3-epoxypropane trimethylammonium chloride is added for reaction for 2-4 hours. Finally, the cationic modified starch is obtained after acid neutralization.

[0015] Comparative Example 1: In the extraction process of kasugamycin fermentation broth, the filtrate ① prepared after the fermentation broth pretreatment and plate frame filtration processes is connected to a storage tank, and different potency elution solutions ② are prepared through ion exchange processes. The elution solutions ② are connected to different storage tanks according to their potencies. When the temperature of the elution solution ② reaches about 40℃, 2% of the polysaccharide additive, cationic starch, is added to the storage tank according to the potency of the elution solution ②. The mixture is uniformly mixed by using an electric mixer to prepare a feed solution ③. The potency and pH value of the feed solution ③ are determined, and the pH value is controlled in the range of 3-5. Then, the 6% kasugamycin aqueous product is prepared by purifying and concentrating the feed solution ③ through membrane filtration.

[0016] Comparative Example 2: In the extraction process of kasugamycin fermentation broth, the filtrate ① prepared after the fermentation broth pretreatment and plate frame filtration processes is connected to a storage tank, and different potency elution solutions ② are prepared through ion exchange processes. The elution solutions ② are connected to different storage tanks according to their potencies. When the temperature of the elution solution ② reaches about 40℃, 8% of the polysaccharide additive, cationic starch, is added to the storage tank according to the potency of the elution solution ②. The mixture is uniformly mixed by using an electric mixer to prepare a feed solution ③. The potency and pH value of the feed solution ③ are determined, and the pH value is controlled in the range of 3-5. Then, the 6% kasugamycin aqueous product is prepared by purifying and concentrating the feed solution ③ through membrane filtration.

[0017] Comparative Example 3: In the extraction process of kasugamycin fermentation broth, the filtrate ① prepared after the fermentation broth pretreatment and plate frame filtration processes is connected to a storage tank, and different potency elution solutions ② are prepared through ion exchange processes. The elution solutions ② are connected to different storage tanks according to their potencies. When the temperature of the elution solution ② reaches about 40℃, 4% of the polysaccharide additive, cationic starch, is added to the storage tank according to the potency of the elution solution ②. The mixture is uniformly mixed by using an electric mixer to prepare a feed solution ③. The potency and pH value of the feed solution ③ are determined, and the pH value is controlled in the range of 3-5. Then, the 6% kasugamycin aqueous product is prepared by purifying and concentrating the feed solution ③ through membrane filtration.

[0018] Comparative Example 4: In the extraction process of kasugamycin fermentation broth, the filtrate ① prepared after the fermentation broth pretreatment and plate frame filtration processes is connected to a storage tank, and different potency elution solutions ② are prepared through ion exchange processes. The elution solutions ② are connected to different storage tanks according to their potencies. When the temperature of the elution solution ② reaches about 40℃, 5% of the polysaccharide additive, cationic starch, is added to the storage tank according to the potency of the elution solution ②. The mixture is uniformly mixed by using an electric mixer to prepare a feed solution ③. The potency and pH value of the feed solution ③ are determined, and the pH value is controlled in the range of 3-5. Then, the 6% kasugamycin aqueous product is prepared by purifying and concentrating the feed solution ③ through membrane filtration.

[0019] Example 5: In the extraction process of kasugamycin fermentation broth, the filtrate 1 prepared after the fermentation broth pretreatment and plate frame filtration process was connected to a storage tank, and different potency elution solutions 2 were prepared by ion exchange process. According to the different potency, the elution solution 2 was connected to different storage tanks. When the temperature of the elution solution 2 reached about 40℃, 5% of glycol was added to the storage tank according to the potency of the elution solution 2, and the mixture was uniformly mixed by using an electric mixer to prepare the liquid 3. The potency and pH value of the liquid 3 were determined, and the pH value was controlled in the range of 3-5. Then, the liquid 3 was purified and concentrated by membrane filtration to prepare a 6% kasugamycin aqueous product.

[0020] Example 6: In the extraction process of kasugamycin fermentation broth, the filtrate 1 prepared after the fermentation broth pretreatment and plate frame filtration process was connected to a storage tank, and different potency elution solutions 2 were prepared by ion exchange process. According to the different potency, the elution solution 2 was connected to different storage tanks. When the temperature of the elution solution 2 reached about 40℃, 5% of polysaccharide aid: starch phosphate (cross-linked starch) was added to the storage tank according to the potency of the elution solution 2, and the mixture was uniformly mixed by using an electric mixer to prepare the liquid 3. The potency and pH value of the liquid 3 were determined, and the pH value was controlled in the range of 3-5. Then, the liquid 3 was purified and concentrated by membrane filtration to prepare a 6% kasugamycin aqueous product.

[0021] Example 7: In the extraction process of kasugamycin fermentation broth, the filtrate 1 prepared after the fermentation broth pretreatment and plate frame filtration process was connected to a storage tank, and different potency elution solutions 2 were prepared by ion exchange process. According to the different potency, the elution solution 2 was connected to different storage tanks. When the temperature of the elution solution 2 reached about 40℃, 5% of polysaccharide aid: oxidized starch was added to the storage tank according to the potency of the elution solution 2, and the mixture was uniformly mixed by using an electric mixer to prepare the liquid 3. The potency and pH value of the liquid 3 were determined, and the pH value was controlled in the range of 3-5. Then, the liquid 3 was purified and concentrated by membrane filtration to prepare a 6% kasugamycin aqueous product.

[0022] Example 8: In the extraction process of kasugamycin fermentation broth, the filtrate 1 prepared after the fermentation broth pretreatment and plate frame filtration process was connected to a storage tank, and different potency elution solutions 2 were prepared by ion exchange process. According to the different potency, the elution solution 2 was connected to different storage tanks. When the temperature of the elution solution 2 reached about 40℃, 5% of polysaccharide aid: enzyme-modified starch (obtained by adding starch to enzyme and enzymatic hydrolysis) was added to the storage tank according to the potency of the elution solution 2, and the mixture was uniformly mixed by using an electric mixer to prepare the liquid 3. The potency and pH value of the liquid 3 were determined, and the pH value was controlled in the range of 3-5. Then, the liquid 3 was purified and concentrated by membrane filtration to prepare a 6% kasugamycin aqueous product.

[0023] Example 9: In the extraction process of kasugamycin fermentation broth, the filtrate ① was prepared after the fermentation broth was pretreated and filtered by plate and frame. Different elution solutions ② with different valences were prepared by ion exchange process. The elution solution ② was added into different storage tanks according to the valence. When the temperature of the elution solution ② reached about 40℃, 5% polysaccharide additive was added into the storage tank according to the valence of the elution solution ②. The mixture was uniformly mixed by using an electric mixer to prepare the liquid ③. The valence and PH value of the liquid ③ were determined. The PH value was controlled in the range of 3-5. Then, the 6% kasugamycin aqueous solution product was prepared by purifying and concentrating the liquid ③ through membrane filtration.

[0024] The kasugamycin aqueous solution without adding polysaccharide additive denatured starch was used as No. 1 product. The kasugamycin aqueous solution with anti-freezing and high re-dissolution rate prepared in Example 1 was used as No. 2 product. The index detection before freezing was carried out to obtain Table 1 and the liquid chromatography detection data graph in Figure Figure 2 Table 2 and the liquid chromatography detection data graph in Figure Figure 4 after freezing and thawing. Table (3) is the re-dissolution effect test comparison results of different additives. Table (3) shows that, except for adding 4%-6% cationic starch prepared by the present application, the rest of the additives all have the problem of light shaking and cannot be completely re-dissolved.

[0025] Figure Figure 3a -3d shows the product performance during freezing and the re-dissolution properties after thawing, Figure 3a Figure shows that the kasugamycin aqueous solution without adding polysaccharide additive was obviously frozen after being frozen at minus 10℃ for 24 hours, while the kasugamycin aqueous solution with polysaccharide additive was partially frozen after being frozen at minus 10℃ for 24 hours and then being frozen at minus 18℃ for 12 hours; Figure 3c Figure -3d shows that the frozen kasugamycin aqueous solutions of No. 1 and No. 2 were thawed. It was found that the re-dissolution rate of the kasugamycin aqueous solution with polysaccharide additive denatured starch was significantly lower than that of the kasugamycin aqueous solution with polysaccharide additive denatured starch. The test product No. 2 of the kasugamycin aqueous solution with polysaccharide additive denatured starch can not be frozen for 24 hours, has the anti-freezing effect, and after being frozen at minus 10℃, it can be re-dissolved within 24 hours at room temperature 18℃, and after being gently shaken, the re-dissolution rate reaches 100%.

[0026] Table (1)

[0027]

[0028] Table (2)

[0029]

[0030] Table (3)

[0031]

Claims

1. A kasugamycin aqueous formulation having improved reconstitution rate, characterized in that, In the spring Lei mycelium fermentation extract liquid is added 4%-6% mass ratio of polysaccharide auxiliary denatured starch;The denatured starch is cationic starch;The preparation steps of cationic starch include: corn starch 100g is added to 500g water and stirred and dispersed, enzyme is added for enzymolysis, the enzymolysis time is 20-30 minutes to obtain the enzymolysis starch liquid;The enzymolysis starch liquid is heated to 40-50℃, then 6-8% mass percentage of lye is added, the temperature is raised to 70-90℃, 2, 3-epoxy propane trimethyl ammonium chloride is added and reacted for 2-4 hours, and finally the post-treatment is carried out to obtain the cationic modified starch.

2. The kasugamycin aqueous formulation with improved reconstitution rate according to claim 1, characterized in that, Specifically, the spring Lei mycelium fermentation liquid is prepared into filtrate ① by pretreatment and plate frame filtration process during the extraction process, and then the filtrate ① is connected to the storage tank to obtain the spring Lei mycelium fermentation stock solution, and then different potency of eluate ② is prepared by ion exchange process, the eluate ② is connected to different storage tanks according to the different potency, when the temperature of the eluate ② reaches 40℃, 4%-6% mass ratio of polysaccharide auxiliary denatured starch is added in the storage tank according to the potency of the eluate ②, and then the mixture is uniformly mixed by using an electric mixer to prepare the material liquid ③, the potency and PH value of the material liquid ③ are determined, the PH value is controlled in the range of 3-5, then the material liquid ③ is further purified and concentrated by membrane filtration to prepare the spring Lei mycelium antifreeze water agent product containing 2% and 6% spring Lei mycelium with polysaccharide auxiliary denatured starch.

3. A process for preparing kasugamycin aqueous formulation with improved reconstitution rate, characterized in that, In the spring Lei mycelium fermentation extract liquid is added 4%-6% mass ratio of polysaccharide auxiliary denatured starch;The denatured starch is cationic starch;The preparation steps of cationic starch include: corn starch 100g is added to 500g water and stirred and dispersed, enzyme is added for enzymolysis, the enzymolysis time is 20-30 minutes to obtain the enzymolysis starch liquid;The enzymolysis starch liquid is heated to 40-50℃, then 6-8% mass percentage of lye is added, the temperature is raised to 70-90℃, 2, 3-epoxy propane trimethyl ammonium chloride is added and reacted for 2-4 hours, and finally the post-treatment is carried out to obtain the cationic modified starch.

4. The preparation process of kasugamycin aqueous agent with improved reconstitution rate according to claim 3, characterized in that, Specifically, the spring Lei mycelium fermentation liquid is prepared into filtrate ① by pretreatment and plate frame filtration process during the extraction process, and then the filtrate ① is connected to the storage tank to obtain the spring Lei mycelium fermentation stock solution, and then different potency of eluate ② is prepared by ion exchange process, the eluate ② is connected to different storage tanks according to the different potency, when the temperature of the eluate ② reaches 40℃, 4%-6% mass ratio of polysaccharide auxiliary denatured starch is added in the storage tank according to the potency of the eluate ②, and then the mixture is uniformly mixed by using an electric mixer to prepare the material liquid ③, the potency and PH value of the material liquid ③ are determined, the PH value is controlled in the range of 3-5, then the material liquid ③ is further purified and concentrated by membrane filtration to prepare the spring Lei mycelium antifreeze water agent product containing 2% and 6% spring Lei mycelium with polysaccharide auxiliary denatured starch.

5. The use of a modified starch additive aid in improving the freeze resistance and resolubility of kasugamycin, characterized by, In the spring Lei mycelium fermentation extract liquid is added 4% -6% mass ratio of polysaccharide adjuvant denatured starch;The denatured starch is cationic starch;The preparation steps of cationic starch include: 100g of corn starch is added to 500g of water and stirred to disperse, enzyme is added for enzymolysis, the enzymolysis time is 20-30 minutes to obtain enzymolysis starch liquid;The enzymolysis starch liquid is heated to 40-50℃, then 6-8% mass fraction of lye is added, the temperature is raised to 70-90℃, 2,3-epoxy propane trimethyl ammonium chloride is added and reacted for 2-4 hours, and finally post-treatment is carried out to obtain cationic modified starch.

6. Use according to claim 5, characterized in that, Specifically, the spring Lei mycelium fermentation liquid is prepared into filtrate ① by pretreatment and plate frame filtration process during extraction, and then the filtrate ① is connected to a storage tank to obtain spring Lei mycelium fermentation stock solution, and then different potency of eluate ② is prepared by ion exchange process, the eluate ② is connected to different storage tanks according to different potencies, when the temperature of the eluate ② reaches 40℃, 4%-6% mass ratio of polysaccharide adjuvant denatured starch is added to the storage tank according to the potency of the eluate ②, and then the mixture is uniformly mixed by using an electric mixer to prepare material liquid ③, the potency and PH value of the material liquid ③ are determined, the PH value is controlled in the range of 3-5, then the material liquid ③ is purified and concentrated by membrane filtration to prepare spring Lei mycelium antifreeze water agent products with 2% and 6% spring Lei mycelium mass concentration and polysaccharide adjuvant denatured starch.

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