Preparation method of chloro-oxytetracycline additive for feed

Abstract

The invention belongs to a feed additive, and particularly relates to a preparation method of a chloro-oxytetracycline additive for feed, which comprises the following steps: step 1, spore preparation: inoculating preserved strains to a slant culture medium, culturing spores under proper temperature and humidity conditions, adding a proper amount of nutrient substances, and culturing the spores under proper temperature and humidity conditions; the method has the beneficial effects that oxytetracycline is released and impurities are removed through acidification and wall breaking of oxalic acid, the ammonia water promotes crystallization by adjusting the pH value, and the production efficiency and the product quality of oxytetracycline can be remarkably improved through combined use of the oxalic acid and the ammonia water; in addition, wastewater generated by using oxalic acid and ammonia water can be treated through a reasonable wastewater treatment process and recycling measures, so that the influence on the environment can be reduced; when the prepared oxytetracycline is used for producing chloro-oxytetracycline, the method has the advantages of raw material cost control, production process optimization, technical innovation and application, environmental protection, sustainability, market competitive advantage and the like.

Description

Technical Field

[0001] This invention pertains to feed additives, specifically to a method for preparing a chlorooxytetracycline additive for use in feed. Background Technology

[0002] Oxytetracycline is an organic compound, a pale yellow crystalline powder, slightly soluble in ethanol, very slightly soluble in water, stable in air, and gradually darkens in color upon exposure to light. Oxytetracycline is an amphoteric compound, capable of forming salts with acids or bases. It is very slightly soluble in water, readily soluble in dilute alkalis and acids. Oxytetracycline salts are easily destroyed and inactivated in alkaline aqueous solutions, but are relatively stable in acidic aqueous solutions. The production of chlorooxytetracycline from oxytetracycline is widely used to treat infections caused by various bacteria, particularly in the veterinary field, where it is frequently used as a feed additive. However, current methods for producing chlorooxytetracycline generate corrosive substances with significant toxic side effects and pose substantial environmental hazards; furthermore, the wastewater produced is difficult to recycle. Summary of the Invention

[0003] To address the problems mentioned in the background section, this invention provides a method for preparing a chlorooxytetracycline additive for animal feed, characterized by raw material cost control, optimized production process, technological innovation and application, and environmental friendliness and sustainability.

[0004] To achieve the above objectives, the present invention provides the following technical solution: a method for preparing chlorooxytetracycline additive for feed, the method comprising the following steps: Step 1: Spore preparation. The preserved strain is inoculated onto an agar slant culture medium, and the spores are cultured under suitable temperature and humidity conditions, with the addition of appropriate nutrients. Step 2: Seed culture. After inoculating the spores into the primary seed tank and culturing them for a period of time, the resulting seed liquid is inoculated into the secondary seed tank for further culture to obtain mycelium. The obtained mycelium is then injected into a fermenter for large-scale liquid fermentation, while adding an appropriate amount of nutrients. Step 3: Acidification and filtration. The fermentation broth is acidified with an acidifying agent to release oxytetracycline in the mycelium and remove impurities from the fermentation broth. The acidified fermentation broth is then filtered by centrifugation to separate the solid and liquid components. Step 4: Decolorization and crystallization. The filtrate is decolorized using resin to remove pigments and organic impurities. The pH of the decolorized filtrate is then adjusted to near the isoelectric point using an alkalizing agent, allowing oxytetracycline to precipitate and crystallize directly from the filtrate. Step 5: Centrifugal drying. The crystallization liquid is sent to a centrifuge for solid-liquid separation to obtain wet oxytetracycline crystals. After rinsing, the wet crystals are dried at an appropriate temperature for a period of time to obtain the dried oxytetracycline product. Step 6: Dissolve oxytetracycline. Add an appropriate amount of oxytetracycline and ethanol into the reactor and stir to promote dissolution. Then add a chlorination reagent to the ethanol solution containing the dissolved oxytetracycline. Step 7: Solid-liquid separation. As the reaction proceeds, chloroxycycline will gradually precipitate in the form of crystals. The crystals are separated into solid and liquid phases using a centrifuge. The resulting solid product is then washed with ethanol. Step 8: Feed mixing. The weighed chloroxycycline is thoroughly mixed with the feed using a feed mixer to ensure that the chloroxycycline is evenly distributed in the feed. Step Nine: Feeding. Feed the animals with the feed containing chloroxycycline, closely observe their health and growth, and promptly identify and address any adverse reactions.

[0005] As a further aspect of the present invention: the culture medium is composed of bran and agar, the spore cultivation temperature needs to be maintained at 36.5℃~36.8℃, the relative humidity is maintained at 55%~60%, and the cultivation time in step one is 90~100h.

[0006] As a further aspect of the present invention: the spores are fermented for 25 to 30 hours in the primary seed tank, the secondary seed tank, and the fermentation tank, and the fermentation temperature is 30°C to 31°C. The nutrients are composed of soluble starch, peptone, and sodium chloride.

[0007] As a further aspect of the present invention: the acidifying agent is oxalic acid, the pH value of the solvent after acidification in step three is 1.6 to 1.9, the centrifuge speed in step three is 2000 rpm to 4000 rpm, and the time is 8 to 12 min.

[0008] As a further aspect of the present invention: the resin is 122# resin, the alkalizing agent is ammonia water, and the pH value is adjusted to 4.6-4.8 in step four.

[0009] As a further aspect of the present invention: in step five, the centrifuge speed is 3000 rpm to 6000 rpm, the time is 5 to 10 min, the drying temperature is 50℃ to 70℃, and the drying time is 0.5 to 1 h.

[0010] As a further aspect of the present invention: the chlorination reagent is sodium hypochlorite, the content ratio of oxytetracycline, ethanol and chlorination reagent is 1:6.4:0.5, the reactor is equipped with a stirring mechanism, the stirring time is 65-90 min, and the temperature inside the reactor is 0℃-7℃.

[0011] As a further aspect of the present invention: in step seven, the centrifuge speed is 4500 rpm to 6000 rpm and the time is 7 to 15 minutes; the ethanol washing time for chloroxycycline is 45 to 60 minutes; and the feed mixer operation time is 30 minutes.

[0012] Compared with existing technologies, the beneficial effects of this invention are as follows: This invention releases oxytetracycline and removes impurities through oxalic acid acidification to break the cell wall, while ammonia water promotes crystallization by adjusting the pH value. The combined use of these two methods can significantly improve the production efficiency and product quality of oxytetracycline. Furthermore, the wastewater generated by the use of oxalic acid and ammonia water can be treated through reasonable wastewater treatment processes and recycling measures, which can reduce the environmental impact. The production of chlorooxytetracycline using the prepared oxytetracycline has many advantages, including raw material cost control, production process optimization, technological innovation and application, environmental protection and sustainability, and market competitiveness. At the same time, the use of ethanol and sodium hypochlorite to produce chlorooxytetracycline has excellent solvent and reactant properties, synergistic effect and reaction control, and environmental protection and sustainability. Detailed Implementation

[0013] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0014] Example This invention provides the following technical solution: a method for preparing chlorooxytetracycline additive for feed, the method comprising the following steps: Step 1: Spore preparation. The preserved strain is inoculated onto an agar slant culture medium, and the spores are cultured under suitable temperature and humidity conditions, with the addition of appropriate nutrients. Step 2: Seed culture. After inoculating the spores into the primary seed tank and culturing them for a period of time, the resulting seed liquid is inoculated into the secondary seed tank for further culture to obtain mycelium. The obtained mycelium is then injected into a fermenter for large-scale liquid fermentation, while adding an appropriate amount of nutrients. Step 3: Acidification and filtration. The fermentation broth is acidified with an acidifying agent to release oxytetracycline in the mycelium and remove impurities from the fermentation broth. The acidified fermentation broth is then filtered by centrifugation to separate the solid and liquid components. Step 4: Decolorization and crystallization. The filtrate is decolorized using resin to remove pigments and organic impurities. The pH of the decolorized filtrate is then adjusted to near the isoelectric point using an alkalizing agent, allowing oxytetracycline to precipitate and crystallize directly from the filtrate. Step 5: Centrifugal drying. The crystallization liquid is sent to a centrifuge for solid-liquid separation to obtain wet oxytetracycline crystals. After rinsing, the wet crystals are dried at an appropriate temperature for a period of time to obtain the dried oxytetracycline product. Step 6: Dissolve oxytetracycline. Add an appropriate amount of oxytetracycline and ethanol into the reactor and stir to promote dissolution. Then add a chlorination reagent to the ethanol solution containing the dissolved oxytetracycline. Step 7: Solid-liquid separation. As the reaction proceeds, chloroxycycline will gradually precipitate in the form of crystals. The crystals are separated into solid and liquid phases using a centrifuge. The resulting solid product is then washed with ethanol. Step 8: Feed mixing. The weighed chloroxycycline is thoroughly mixed with the feed using a feed mixer to ensure that the chloroxycycline is evenly distributed in the feed. Step Nine: Feeding. Feed the animals with the feed containing chloroxycycline, closely observe their health and growth, and promptly identify and address any adverse reactions.

[0015] Specifically, the culture medium consists of bran and agar, the spore culture temperature needs to be maintained at 36.5℃~36.8℃, the relative humidity needs to be maintained at 55%~60%, and the culture time in step one is 90~100h.

[0016] Specifically, the spores are fermented for 25 to 30 hours in the primary seed tank, secondary seed tank, and fermentation tank, at a temperature of 30°C to 31°C, and the nutrients consist of soluble starch, peptone, and sodium chloride.

[0017] Specifically, the acidifying agent is oxalic acid, the pH value of the solvent after acidification in step three is 1.6 to 1.9, the centrifuge speed in step three is 2000 rpm to 4000 rpm, and the time is 8 to 12 min.

[0018] Specifically, the resin is 122# resin, the alkalizing agent is ammonia, and the pH value is adjusted to 4.6-4.8 in step four.

[0019] Specifically, in step five, the centrifuge speed is 3000 rpm to 6000 rpm, the time is 5 to 10 minutes, the drying temperature is 50℃ to 70℃, and the drying time is 0.5 to 1 hour.

[0020] Specifically, the chlorination reagent is sodium hypochlorite, and the content ratio of oxytetracycline, ethanol and chlorination reagent is 1:6.4:0.5. The reactor is equipped with a stirring mechanism, the stirring time is 65-90 minutes, and the temperature inside the reactor is 0℃-7℃.

[0021] Specifically, in step seven, the centrifuge speed is 4500 rpm to 6000 rpm for 7 to 15 minutes, the ethanol washing time for chloroxycycline is 45 to 60 minutes, and the feed mixer runs for 30 minutes.

[0022] The working principle and usage process of this invention are as follows: First, the preserved microbial strain is inoculated onto a slant culture medium. Spores are cultured under suitable temperature and humidity conditions, with appropriate nutrients added. Then, the spores are inoculated into a primary seed tank and cultured for a period of time. The resulting seed liquid is then inoculated into a secondary seed tank for further culture to obtain mycelium. The obtained mycelium is then injected into a fermenter for large-scale liquid fermentation, with appropriate nutrients added. The fermentation broth is then acidified with an acidifying agent to release oxytetracycline from the mycelium and remove impurities. The acidified fermentation broth is then filtered by centrifugation for solid-liquid separation. The filtrate is then decolorized with resin to remove pigments and organic impurities. The decolorized filtrate is then adjusted to near its isoelectric point using an alkalizing agent, causing oxytetracycline to precipitate and crystallize directly from the filtrate. The crystallized liquid is then sent to a centrifuge. Solid-liquid separation was performed to obtain wet oxytetracycline crystals. After rinsing, the wet crystals were dried at an appropriate temperature for a period of time to obtain dried oxytetracycline. An appropriate amount of oxytetracycline and ethanol were added to the reactor and stirred to promote dissolution. Then, a chlorination reagent was added to the ethanol solution containing dissolved oxytetracycline. As the reaction proceeded, chlorooxytetracycline gradually precipitated in the form of crystals. The crystals were separated into solid and liquid components using a centrifuge to remove the mother liquor, unreacted raw materials, and byproducts. The obtained solid product was then washed with ethanol to remove impurities adhering to the crystal surface. The weighed chlorooxytetracycline was then thoroughly mixed with feed using a feed mixer to ensure uniform distribution of chlorooxytetracycline in the feed. Finally, the feed supplemented with chlorooxytetracycline was fed to the animals. The health and growth of the animals were closely observed, and any adverse reactions were promptly detected and addressed.

[0023] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing a chlorooxytetracycline additive for feed, characterized in that: The method includes the following steps: Step 1: Spore preparation. The preserved strain is inoculated onto an agar slant culture medium, and the spores are cultured under suitable temperature and humidity conditions, with the addition of appropriate nutrients. Step 2: Seed culture. After inoculating the spores into the primary seed tank and culturing them for a period of time, the resulting seed liquid is inoculated into the secondary seed tank for further culture to obtain mycelium. The obtained mycelium is then injected into a fermenter for large-scale liquid fermentation, while adding an appropriate amount of nutrients. Step 3: Acidification and filtration. The fermentation broth is acidified with an acidifying agent to release oxytetracycline in the mycelium and remove impurities from the fermentation broth. The acidified fermentation broth is then filtered by centrifugation to separate the solid and liquid components. Step 4: Decolorization and crystallization. The filtrate is decolorized using resin to remove pigments and organic impurities. The pH of the decolorized filtrate is then adjusted to near the isoelectric point using an alkalizing agent, allowing oxytetracycline to precipitate and crystallize directly from the filtrate. Step 5: Centrifugal drying. The crystallization liquid is sent to a centrifuge for solid-liquid separation to obtain wet oxytetracycline crystals. After rinsing, the wet crystals are dried at an appropriate temperature for a period of time to obtain the dried oxytetracycline product. Step 6: Dissolve oxytetracycline. Add an appropriate amount of oxytetracycline and ethanol into the reactor and stir to promote dissolution. Then add a chlorination reagent to the ethanol solution containing the dissolved oxytetracycline. Step 7: Solid-liquid separation. As the reaction proceeds, chloroxycycline will gradually precipitate in the form of crystals. The crystals are separated into solid and liquid phases using a centrifuge. The resulting solid product is then washed with ethanol. Step 8: Feed mixing. The weighed chloroxycycline is thoroughly mixed with the feed using a feed mixer to ensure that the chloroxycycline is evenly distributed in the feed. Step Nine: Feeding. Feed the animals with the feed containing chloroxycycline, closely observe their health and growth, and promptly identify and address any adverse reactions.

2. The method for preparing a chlorooxytetracycline additive for feed according to claim 1, characterized in that: The culture medium consists of bran and agar. The spore culture temperature needs to be maintained at 36.5℃~36.8℃, the relative humidity needs to be maintained at 55%~60%, and the culture time in step one is 90~100h.

3. The method for preparing a chlorooxytetracycline additive for feed according to claim 1, characterized in that: The spores are fermented for 25 to 30 hours in the primary seed tank, secondary seed tank, and fermentation tank, at a temperature of 30°C to 31°C. The nutrients consist of soluble starch, peptone, and sodium chloride.

4. The method for preparing a chlorooxytetracycline additive for feed according to claim 1, characterized in that: The acidifying agent is oxalic acid, the pH value of the solvent after acidification in step three is 1.6 to 1.9, the centrifuge speed in step three is 2000 rpm to 4000 rpm, and the time is 8 to 12 min.

5. The method for preparing a chlorooxytetracycline additive for feed according to claim 1, characterized in that: The resin is 122# resin, the alkalizing agent is ammonia, and the pH value is adjusted to 4.6-4.8 in step four.

6. The method for preparing a chlorooxytetracycline additive for feed according to claim 1, characterized in that: In step five, the centrifuge speed is 3000 rpm to 6000 rpm, the time is 5 to 10 minutes, the drying temperature is 50℃ to 70℃, and the drying time is 0.5 to 1 hour.

7. The method for preparing a chlorooxytetracycline additive for feed according to claim 1, characterized in that: The chlorinating agent is sodium hypochlorite, and the content ratio of oxytetracycline, ethanol and chlorinating agent is 1:6.4:0.

5. The reactor is equipped with a stirring mechanism, the stirring time is 65-90 min, and the temperature inside the reactor is 0℃-7℃.

8. The method for preparing a chlorooxytetracycline additive for feed according to claim 1, characterized in that: In step seven, the centrifuge speed is 4500 rpm to 6000 rpm and the time is 7 to 15 minutes; the ethanol washing time for chloroxycycline is 45 to 60 minutes; and the feed mixer operation time is 30 minutes.