WO2021203606A1 - Preparation method for compound amino acid injection for livestock - Google Patents

Abstract

A preparation method for a compound amino acid injection for livestock, comprising: boiling water for injection for sterilization, and then adding an antioxidant ethylenediaminetetraacetic acid disodium salt and an antioxidant sodium bisulfite at 75-95°C under the protection of a CO ₂ gas flow; adding 15 amino acids at 70-100°C under the protection of CO ₂ gas, then adding tryptophan, tyrosine, and cysteine hydrochloride at 60-70°C, finally adding γ-aminobutyric acid at 20-35°C, regulating the pH value, performing decolorization, etc. The problem of poor stability of amino acid injections for livestock is solved. An amino acid injection for livestock is prepared by multi-step feeding; tryptophan, tyrosine, and cysteine hydrochloride are added at 60-70°C under the protection of CO₂; thus, the stability of the tryptophan and the tyrosine is effectively improved and the tryptophan and the tyrosine are prevented from being oxidized; the prepared injection is suitable for disease treatment of livestock and body recovery after diseases.

Description

Preparation method of compound amino acid injection for animal use Technical field

The invention belongs to the technical field of animal husbandry and veterinary medicine, and relates to a preparation method of animal injection, in particular to a preparation method of animal compound amino acid injection.

Background technique

At present, the supplementary liquid used to enhance the physical fitness of animals mainly uses glucose injection, sodium chloride injection and glucose sodium chloride injection. These three injections mainly play the role of supplementing water and energy, and regulating the balance of body electrolytes, but they cannot improve the nutritional status of the animal body and supplement the amino acids required by the animal body, and thus cannot effectively regulate the animal's feed intake, especially for the weak During the onset of the disease, the animals need to supplement amino acids to improve their nutrition to promote the recovery of the animals.

In general, frail and diseased animals mostly show symptoms such as decreased appetite, reduced feed intake, and physical weakness. Due to the above symptoms, a large amount of nutrients in their bodies are lost over time. If the above symptoms are not improved in time, the animals will grow slowly or even die, which in turn will bring greater economic losses to the breeding industry.

The Chinese invention patent with the patent number 201210000911.5 discloses a technical solution of compound amino acid injection. The technical solution is mainly used for the treatment of animal diseases and the restoration of the body during the recovery period after the disease, supplementing the essential and non-essential amino acids required by the animal body. , Adjuvant treatment of diseases and post-ill recovery period to enhance physical fitness and increase animal feed intake.

This technical solution has a certain effect on the treatment of animal diseases and the restoration of the body in the recovery period after the disease. Although the invention patent discloses the preparation method of compound amino acid injection, the prepared animal compound amino acid injection still has the defect of poor stability. The specific reasons for the above defects are as follows:

In the preparation process of the existing animal compound amino acid injection, under the protection of carbon dioxide, tyrosine and other amino acids are added to the basic liquid at 75-95°C. However, because tyrosine is unstable at high temperatures and is prone to oxidative deterioration, the stability of compound amino acid injections for livestock is reduced, which is not conducive to the preparation and storage of compound amino acid injections for livestock.

In summary, the existing animal compound amino acid injections have poor amino acid stability, which is not conducive to the storage of animal compound amino acid injections.

Summary of the invention

The purpose of the present invention is to provide a method for preparing a compound amino acid injection for livestock, so as to solve the problem of poor stability of each amino acid in the injection for livestock.

In order to achieve the above objective, the technical solution adopted by the present invention is:

A preparation method of compound amino acid injection for animal use includes the following steps in sequence:

Step 1) Prepare the basic solution:

Take 600-800 parts by weight of water for injection and boil it for sterilization, pass in CO ₂ gas, and control the water temperature at 75-100 ℃ at the same time, under the _{protection of CO 2} gas flow, add the antioxidant disodium edetate and antioxidant Sodium bisulfite, dissolve to prepare basic liquid A;

Step 2) Prepare the intermediate solution:

Under the _{protection of CO 2} gas, add group B amino acids to basic solution A, dissolve at 70-100°C, and then naturally cool down to 60-70°C to prepare solution C;

Among them, group B amino acids are calculated in parts by weight, including: 1.00 to 4.00 parts of arginine, 0.80 to 2.54 parts of histidine, 1.50 to 2.88 parts of leucine, 0.50 to 2.00 parts of isoleucine, and 1.00 to 4.50 parts Lysine, 1.00 to 2.96 parts of phenylalanine, 0.80 to 2.11 parts of threonine, 0.50 to 2.50 parts of valine, 0.50 to 2.10 parts of methionine, 1.00 to 3.45 parts of glycine, 0.80 to 3.00 parts of alanine Acid, 0.50 to 2.40 parts of proline, 0.25 to 1.50 parts of serine, 0.50 to 1.25 parts of aspartic acid, 0.80 to 2.25 parts of glutamic acid;

Under the _{protection of CO 2} gas, add group D amino acids to solution C, dissolve at 60～70℃, and naturally cool to 20～35℃ to prepare solution E;

Among them, the amino acids of group D are calculated in parts by weight, including: 0.20 to 0.70 parts of tryptophan, 0.05 to 0.15 parts of tyrosine, and 0.20 to 1.00 parts of cysteine hydrochloride;

Under the _{protection of CO 2} gas, add 0.50 to 2.00 parts of γ-aminobutyric acid to solution E. After dissolving at 20 to 35°C, adjust the pH to 5.7 to 7.0, and dilute to 1000 parts by volume to obtain intermediate solution F;

Among them, the ratio of parts by weight to parts by volume is kg:L;

Step 3) Preparation of compound amino acid injection for animal use:

Under the _{protection of CO 2} gas, active carbon is added to the intermediate solution F, decolorized at 20-35° C., and filtered to prepare the compound amino acid injection for livestock.

As a limitation of the present invention, the prepared animal compound amino acid injection is sterilized.

As a further limitation of the present invention, before sterilization, under the _{protection of CO 2} gas, the animal compound amino acid injection is filled.

As a limitation of the present invention, in step 3), the weight-volume ratio of the activated carbon to the intermediate solution F is 0.5-1 kg:1000L.

As a further limitation of the present invention, in step 3), the decolorization temperature is 20 to 35° C. and the time is 30 to 40 min.

As a further limitation of the present invention, in step 3), filtration includes coarse filtration and fine filtration in sequence;

Fine filtration is a microporous filter element filtration;

The pore size of the microporous filter element is 0.45μm, 0.22μm or 0.20μm.

As a further limitation of the present invention, the residual oxygen content in the container during the filling process is less than or equal to 3%.

As a further limitation of the present invention, the sterilization temperature is 110-121°C, and the time is 15-40 min.

Compared with the prior art, the present invention improves the content and stability of amino acids in compound amino acid injections for livestock by changing the process parameters, and the details are as follows:

First, the present invention adopts stepwise feeding to prepare animal amino acid injection, and simultaneously adds tryptophan, tyrosine, and cysteine hydrochloride at 60-70°C under the protection of _{CO 2 to reduce the feeding temperature of tyrosine} , While ensuring that the solubility of tyrosine does not decrease, it effectively prevents the oxidation of tryptophan and tyrosine, and improves the stability of compound amino acid injections for livestock;

Second, the addition of cysteine hydrochloride together with tyrosine and tryptophan can protect tryptophan and tyrosine from being oxidized, and effectively improve the stability of tryptophan and tyrosine, and the level of tryptophan is 60%. The solubility reaches the best at ～70℃, which not only increases the content of tryptophan and tyrosine in animal amino acid injection, but also improves the stability of tyrosine and tryptophan.

Third, by changing the feeding conditions to prepare animal amino acid injections, the dissolution time is shortened, thereby reducing the preparation time and improving the production efficiency.

The animal compound amino acid injection prepared by the invention is suitable for body repair during the treatment of animal diseases and during the recovery period after the disease.

Detailed ways

The present invention will be further described in detail below through specific embodiments. It should be understood that the described embodiments are only used to explain the present invention and do not limit the present invention.

Example 1 Preparation method of compound amino acid injection for livestock

This embodiment is a preparation method of compound amino acid injection for animals. Taking 1m3 compound injection for animals as an example, the specific preparation method includes the following steps in sequence:

Step 1) Prepare the basic solution:

Take 1000L of water for injection and sterilize it by boiling for 40min, then set aside;

Weigh 0.89kg of sodium bisulfite and 0.10kg of disodium edetate for use;

Take 600L of water for injection and place it in a concentrated _{mixing tank, pass CO 2} gas flow, and at the same time control the water temperature at 95°C, add 0.89kg disodium ethylenediaminetetraacetic acid and 0.10kg sodium bisulfite under the protection of _{CO 2 gas flow, and stir} Make it completely dissolve, and prepare the base liquid A.

Among them, sodium bisulfite has the effect of preventing the oxidation of tryptophan and tyrosine. Therefore, adding sodium bisulfite before adding each amino acid has a better antioxidant effect. Disodium ethylenediaminetetraacetate can be combined with the raw materials. The introduced metal ions perform chelation to prevent the metal ions from catalyzing the oxidation of amino acids. Therefore, in this embodiment, the sodium bisulfite and disodium edetate are dissolved before adding each amino acid.

Tryptophan in compound amino acid injection is a component that is easy to oxidize and decompose. Other amino acids may also be oxidized and decomposed. During production and storage, the stability of amino acids is greatly affected by the oxygen content in the liquid. The higher the oxygen content, the more the amino acid is oxidized and decomposed, the worse the stability, and the darker the color of the drug solution. Therefore, the inert gas protection is used at the beginning of the preparation, which can effectively reduce the oxygen content in the drug solution, so that the amino acid can be sterilized and sterilized later. More stable during storage.

Step 2) Prepare the intermediate solution:

Weigh 1.00kg arginine, 2.54kg histidine, 2.54kg leucine, 1.52kg isoleucine, 3.33kg lysine, 2.96kg phenylalanine, 0.80kg threonine, 2.50kg valine Acid, 1.79kg methionine, 3.24kg glycine, 0.80kg alanine, 2.40kg proline, 0.25kg serine, 0.50kg aspartic acid, 1.34kg glutamic acid, collectively referred to as group B amino acids;

Under the _{protection of CO 2} airflow, add the weighed group B amino acids to the basic liquid A at a temperature of 100 ℃, stir to dissolve to obtain solution C, and the temperature of solution C is naturally cooled to 70 ℃;

Weigh 0.32kg tryptophan, 0.15kg tyrosine, 0.76kg cysteine hydrochloride, collectively referred to as group D amino acids

Under the _{protection of CO 2} gas flow, add group D amino acids to solution C at a temperature of 70°C, stir to dissolve to obtain solution E, and solution E is naturally cooled to 30°C;

Under the _{protection of CO 2} gas flow, add 0.89kg of γ-aminobutyric acid to solution E at a temperature of 30℃, stir to dissolve, add water for injection to nearly 1000L, and then adjust the pH value to 6.0 with a mass concentration of 20% sodium hydroxide solution , The final volume was adjusted to 1000L, and the intermediate solution F was prepared.

The dissolution time of amino acids decreases with the increase of temperature. When the temperature is 100℃, the dissolution time is the shortest, and when the temperature is higher than 70℃, most of the amino acid content changes no longer change significantly.

However, due to the instability of tyrosine and tryptophan, the content of tyrosine and tryptophan decreases drastically as the temperature rises. The soluble content of tyrosine and tryptophan in the solution at 60～70℃ is relatively high (the content of tryptophan is relatively high). At 100°C, it increased by 7.4 to 9.0%, and the tyrosine content also increased). Considering factors such as content and dissolution time, each amino acid is added step by step, adding arginine, histidine, leucine, isoleucine, lysine, phenylalanine, threonine, valine, Methionine, glycine, alanine, proline, serine, aspartic acid, and glutamic acid are dissolved at 70～100℃, tryptophan and tyrosine are dissolved at 60～70℃, At the same time, it dissolves cysteine hydrochloride to reduce the oxidation of tryptophan and tyrosine in the process of dissolving.

Because γ-aminobutyric acid has a high solubility at 60-100°C, but it is unstable, and it is easily soluble in water, so the best effect is to dissolve γ-aminobutyric acid at 20-35°C.

During the preparation of the compound amino acid injection, the pH value is controlled at 5.7-7.0, the content of each amino acid is controlled at 80.0-120.0%, the osmolality is not much different, and the amino acid can be stably stored in the solution.

Step 3) Preparation of compound amino acid injection for animal use:

Under the _{protection of CO 2} gas, add 1 kg of activated carbon to the intermediate solution F at a temperature of 30 ℃, decolorize for 30 minutes, filter through coarse filtration and fine filtration with a 0.45 μm microporous filter element to obtain solution G, which is a compound amino acid injection for livestock liquid.

Adding 0.05-0.1% activated carbon to decolorize, the content of each amino acid will decrease to varying degrees. This is because as the amount of activated carbon increases, the adsorbed amount of amino acids will increase, especially tryptophan and tyrosine. When the added amount is 0.05%, the activated carbon has an adsorption effect on amino acids, but after the activated carbon is adsorbed and then sterilized, the solution has good clarity and the sample is stable. In order to improve product quality, 0.05-0.1% activated carbon is added to the preparation process, and the amount of tryptophan and tyrosine raw materials that have a greater impact on the adsorption is appropriately increased during feeding.

Step 4) Prepare bottled animal compound amino acid injection:

Under the _{protection of CO 2} gas, fill solution G into a 1000 mL infusion bottle, sterilize at 115°C for 30 minutes, and pass the lamp inspection to prepare a bottled animal compound amino acid injection;

The residual oxygen content in the infusion bottle has been controlled below 3.0% during the entire filling process.

During the filling process, the residual oxygen content in the infusion bottle is controlled below 3.0%, which can effectively improve the stability of the amino acids in the injection.

Example 2-6 Preparation method of compound amino acid injection for animal use

Examples 2 to 6 are respectively a preparation method of a compound amino acid injection for animals, which is basically the same as the preparation method in Example 1, except that the amount of each amino acid raw material and the parameters in each step are different. Taking 1000L animal compound injection as an example, the amount of specific active ingredient raw materials is weighed according to the amount of each active ingredient raw material given in Table 1. The specific active ingredient raw materials are shown in Table 1; each of the examples Various process parameters in the steps, the specific parameters are shown in Table 2:

Table 1 List of the dosage of each active ingredient raw material

Table 2 List of various process parameters of Examples 2 to 6

The specific conditions of the content of each amino acid in the compound amino acid injection for livestock prepared in Examples 1 to 6 and the content of each amino acid in the existing compound amino acid injection for livestock are shown in Table 3:

Table 3 List of quality test results of compound amino acid injections for livestock prepared in Examples 1 to 6

It can be seen from Table 3 that the color of the compound amino acid injections for animals prepared in Examples 1 to 6 has changed from slightly yellow to colorless and transparent compared with the existing compound amino acid injections for animals, and compared with the existing compound amino acid injections for animals. The liquid has been improved, and other inspection related items have all been qualified.

Example 7 Quality Inspection of Compound Amino Acid Injection for Animals

In this example, the compound amino acid injection for animals prepared by the method in Example 1 was used for quality inspection. The specific detection methods are as follows:

The accelerated test is placed for 6 months at a temperature of 40±2℃ and a relative humidity of 75±5%, and samples are taken at the end of the 0th, 1st, 2nd, 3rd and 6th months of the test period. The quality standards of "National Food and Drug Administration Drug Standards" WS1～(X～324)～2003Z have been tested, and the specific testing results are shown in Table 4:

Table 4 Quality test results of compound amino acid injection for livestock

It can be seen from Table 4 that the animal compound amino acid injection prepared in Example 1 has undergone an accelerated test, with no significant changes in various indicators, and the sample has good quality stability and good stability. Therefore, the preparation method of the present invention can prepare more Stable compound amino acid injection for livestock.

Example 8 Comparison of stability of compound amino acid injection for livestock

According to the quality inspection method of Example 7, the animal compound amino acid injection prepared by the preparation method of Examples 1 to 6 and the existing animal compound amino acid injection were placed for 24 months, and the quality test was compared. The specific test The results are as follows:

Table 5 Comparison results of quality test of compound amino acid injection for livestock

It can be seen from Table 5 that the contents of tryptophan, proline, γ-aminobutyric acid, and tyrosine all decreased after the existing compound amino acid injections for livestock were placed for 24 months. Compound amino acid injection is more stable, and the content does not change much. The content of each amino acid in the animal compound amino acid injection prepared in Examples 1 to 6 is higher than the existing animal compound amino acid injection, which is stable within 24 months, has controllable quality, and is safe and reliable.

Example 9 Application of Compound Amino Acid Injection for Animals

In this example, the animal compound amino acid injection prepared by any of the preparation methods of Examples 1 to 6 is applied to test animals, and the specific method is as follows:

a1) Application in the treatment of bovine viral diarrhea

Bovine viral diarrhea is more susceptible to young cows, and generally exhibits mild symptoms, but sometimes it breaks out suddenly, and the whole group shows severe symptoms. Acute clinical symptoms include sudden fever, body temperature rising to 40-42°C, leukopenia, lack of food or refusal to eat, rumination ceases, rapid breathing and heartbeat, cough, runny nose, oral mucosa flushing, increased saliva, and subsequent erosions and diarrhea Like water, it lasts for several days, with bubbles and blood mixed in the stool. In severe cases, they die of dehydration and exhaustion. The course of the disease is 1 to 3 weeks, and the morbidity and mortality of calves can be greater than 90%. The clinical symptoms of chronic type are not obvious, and the sick cattle show slow growth and development, weight loss, and continuous or intermittent diarrhea. The course of the disease is 2 to 6 months.

The compound amino acid injection for animal use prepared by any of the preparation methods in Examples 1 to 6 was used for clinical trials:

Holstein cattle in Zhonghe Xingmu of Laixi City had symptoms of diarrhea. It was confirmed by the local veterinarian that it was caused by infection with a bovine virus. The plan shown in Table 6 was combined with lincomycin hydrochloride (use amount is 3g/head). ) For treatment.

Table 6 Application of Compound Amino Acid Injection for Animals in Bovine Viral Diarrhea

It can be seen from Table 6 that the compound amino acid injection for livestock has the effects of improving the spirit, increasing appetite, and promoting physical recovery of frail cows after diarrhea, and can be used for adjuvant treatment of frail cows. However, the compound amino acid injections for livestock prepared in Examples 1 to 6 are better than the existing compound amino acid injections for livestock in recovering frailty after diarrhea in cattle.

a2) Application in the treatment of canine parvovirus

Dogs of different ages, genders, and breeds can develop canine parvovirus disease throughout the year (mainly puppies, especially puppies before and after weaning are most susceptible). The clinical symptoms of this disease include depression, loss of appetite, severe vomiting, diarrhea, and blood in the stool. In severe cases, the dog will die due to imbalance of water and electrolyte balance, loss of nutrients in the body and acidosis. The morbidity rate is 20-100%, and the mortality rate is 50-100%. During the onset of the sick dog, fasting and drinking are required. Therefore, the key to rehydration, energy and nutrients is the key to the sick dog. Otherwise, the sick dog is prone to hypoproteinemia and edema. Due to frequent vomiting, oral drugs should not be used. Infusion therapy is the key to treatment. , Timely replenishment of energy, protein and nutrients required by the animal body is the top priority.

The compound amino acid injection for animal use prepared by any of the preparation methods in Examples 1 to 6 was used for clinical trials:

The Beagle dog of Beijing Amerseys Biotechnology Co., Ltd. has diarrhea symptoms. After the clinical symptoms and the parvovirus test paper, the diagnosis is parvovirus disease. The treatment plan is the plan shown in Table 7 and the 10mg/kg antibiotic sulfate card. Namycin and 1mL of ATP～2Na are combined for treatment.

Table 7 Application of Compound Amino Acid Injection for Animals on Canine Parvovirus

It can be seen from Table 7 that the compound amino acid injection for animals can improve the nutritional status of parvovirus-infected dogs, promote the regression of symptoms, and shorten the recovery time. It can be used for the auxiliary treatment of parvovirus-infected dogs. Animal compound amino acid injection can make dogs timely supplement nutrition after illness. After injection, the dog's appetite will be significantly increased, and the weight will be restored. It will help to resolve symptoms such as vomiting and dehydration, and can reduce the mortality of sick dogs. However, the compound amino acid injections for animals prepared in Examples 1 to 6 have better recovery effects on sick dogs than the existing compound amino acid injections for animals.

It should be noted that Embodiments 1 to 6 are only preferred embodiments of the present invention, and are not intended to limit other forms of the present invention. Any person familiar with the profession may use the above technical content as inspiration to make changes or modifications. They are equivalent embodiments with equivalent changes, but simple modifications, equivalent changes and modifications made to the above embodiments without departing from the technical essence of the claims of the present invention still fall within the protection scope of the claims of the present invention.

Claims (8)

1. A preparation method of compound amino acid injection for animals, characterized in that the preparation method comprises the following steps in sequence:

   Step 1) Prepare the basic solution:

   Take the water for injection and boil it for sterilization, pass in CO ₂ gas, and control the water temperature at 75-100 ℃ at the same time, under the _{protection of CO 2} gas flow, add the antioxidant disodium edetate and the antioxidant sodium bisulfite to dissolve , Prepare the basic liquid A;

   Step 2) Prepare the intermediate solution:

   Under the _{protection of CO 2} gas, add group B amino acids to basic solution A, dissolve at 70-100°C, and then lower the temperature to 60-70°C to prepare solution C;

   Among them, group B amino acids include arginine, histidine, leucine, isoleucine, lysine, phenylalanine, threonine, valine, methionine, glycine, alanine , Proline, serine, aspartic acid and glutamic acid;

   Under the _{protection of CO 2} gas, add group D amino acids to solution C, dissolve at 60～70℃, and lower the temperature to 20～35℃ to prepare solution E;

   Among them, group D amino acids include tryptophan, tyrosine and cysteine hydrochloride;

   Under the _{protection of CO 2} gas, add γ-aminobutyric acid to solution E, and after dissolving at 20～35℃, adjust the pH value to 5.7～7.0 and constant volume to obtain intermediate solution F;

   Step 3) Preparation of compound amino acid injection for animal use:

   Under the _{protection of CO 2} gas, the intermediate solution F is added with activated carbon, decolorized and filtered to prepare the animal compound amino acid injection.
2. The method for preparing animal compound amino acid injection according to claim 1, wherein the prepared animal compound amino acid injection is sterilized.
3. The method for preparing animal compound amino acid injection according to claim 2, characterized in that, before sterilization, the animal compound amino acid injection is filled under the protection of _{CO 2 gas.}
4. A method for preparing animal compound amino acid injection according to any one of claims 1 to 3, characterized in that, in step 3), the weight-volume ratio of activated carbon to intermediate solution F is 0.5-1 kg:1000L.
5. The method for preparing animal compound amino acid injection according to claim 4, characterized in that, in step 3), the decolorization temperature is 20-35°C and the time is 30-40 min.
6. The method for preparing animal compound amino acid injection according to claim 4, wherein in step 3), the filtration includes coarse filtration and fine filtration in sequence.
7. The method for preparing animal compound amino acid injection according to claim 3, wherein the residual oxygen content in the container during the filling process is ≤3%.
8. The method for preparing animal compound amino acid injection according to claim 2 or 3, wherein the sterilization temperature is 110-121°C and the time is 15-40 min.