Veterinary antidiarrheal powder as well as preparation method and content determination method thereof

Abstract

The invention discloses veterinary antidiarrheal powder which is prepared from the following components in parts by weight: 1-20 parts of berberine hydrochloride, 1-5 parts of phenyl salicylate, 10-30 parts of montmorillonite, 1-30 parts of yeast and 1-30 parts of calcium carbonate, the prescription design is advanced and reasonable, the veterinary antidiarrheal powder can effectively resist bacteria and inflammation and can quickly stop diarrhea, meanwhile, various nutritional ingredients contained in the yeast are utilized to ensure nutrient absorption of diseased animals, and the effect of preventing and treating the diarrhea is achieved. The feed can effectively treat intestinal diseases of livestock, can be uniformly mixed with feed for feeding, can be used together with machinery, is convenient to use in a farm, and greatly solves the problem of labor cost of drug use in the farm. The invention also discloses a method for determining the contents of the main components berberine hydrochloride and phenyl salicylate in the veterinary antidiarrheal powder, the specificity is strong, the precision is good, the accuracy is high, the method can be used for quality control of the veterinary antidiarrheal powder, and the stability of the product quality and the safety and effectiveness of veterinary drugs are ensured.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical preparations, relates to an antidiarrhea powder for animals, and also relates to a preparation method and a content determination method thereof. Background Art

[0002] Berberine, also known as berberine, is extracted from Coptis chinensis and can also be extracted from natural medicines such as Tripterygium wilfordii and Phellodendron chinense. It is a natural quaternary ammonium isoquinoline alkaloid. Its hydrochloride is mainly used as an over-the-counter drug for the treatment of intestinal infections, with the characteristics of few adverse reactions.

[0003] Veterinary drugs currently include berberine hydrochloride tablets, including two specifications: 0.1g and 0.5g. However, the dosage is large when used in large animals. For example, if a tablet with a specification of 0.1g is used for internal administration, it can be taken for horses and cattle 20 to 50 tablets, camels 30 to 60 tablets, and sheep and pigs 5 to 10 tablets; if a tablet with a specification of 0.5g is used for internal administration, it can be taken for horses and cattle 4 to 10 tablets, camels 6 to 12 tablets, and sheep and pigs 1 to 2 tablets. In intensive and large-scale farms, it is extremely costly to accurately administer tablets. Summary of the invention

[0004] In view of this, the purpose of the present invention is to provide an veterinary antidiarrhea powder with advanced and reasonable prescription design, which can effectively treat livestock intestinal diseases, can be mixed with feed and fed, and can be used with machinery, which greatly solves the labor cost problem of drug use in farms.

[0005] After research, the present invention provides the following technical solutions:

[0006] 1. An antidiarrhea powder for animals, composed of the following components by weight: 1-20 parts of berberine hydrochloride, 1-5 parts of phenyl salicylate, 10-30 parts of montmorillonite, 1-30 parts of yeast, and 1-30 parts of calcium carbonate.

[0007] Preferably, the veterinary antidiarrhea powder consists of the following components by weight: 20 parts of berberine hydrochloride, 2 parts of phenyl salicylate, 30 parts of montmorillonite, 18 parts of yeast, and 30 parts of calcium carbonate.

[0008] In the above-mentioned technical scheme of veterinary antidiarrhea powder, berberine hydrochloride is an antibacterial drug with a broad-spectrum antibacterial effect. It has an antibacterial effect on a variety of Gram-positive and Gram-negative bacteria in vitro, among which it has a stronger effect on hemolytic streptococci, Staphylococcus aureus, Vibrio cholerae, meningococci, Shigella and Salmonella typhi; phenyl salicylate is an analgesic and bactericidal agent with good anti-inflammatory effect, which can reduce inflammatory response and have a bactericidal effect at the same time; montmorillonite has a layered structure and non-uniform charge distribution, which has a fixing and inhibitory effect on viral pathogens in the digestive tract and the toxins they produce, has a covering ability on the digestive tract mucosa, can form a protective layer on the surface of the gastrointestinal mucosa, and by combining with mucus glycoproteins, it can repair and improve the gastrointestinal mucosa's defense function against pathogenic factors, and mainly plays an antidiarrheal effect; yeast belongs to a vitamin drug, containing thiamine, riboflavin, niacin, and also contains vitamin B6 and vitamin B 12 , folic acid, inositol, invertase, maltase, etc., all of which are important components of the enzyme system in the body, participate in the metabolism and biotransformation of sugar, protein, fat, etc. in the body, and promote the intestinal absorption of nutrients; calcium carbonate is mainly a drug carrier, which can neutralize or buffer gastric acid and protect the effective ingredients of other drugs. The veterinary antidiarrhea powder of the present invention belongs to a compound antidiarrhea medicine, which can effectively resist bacteria and inflammation, and can quickly stop diarrhea. At the same time, it uses various nutrients contained in yeast to ensure the nutrient absorption of sick animals, quickly restore their digestive function, and can effectively treat intestinal diseases of livestock.

[0009] 2. The preparation method of the veterinary antidiarrhea powder comprises the following steps: mixing phenyl salicylate and montmorillonite, adding berberine hydrochloride and yeast, mixing, and mixing the obtained mixture with calcium carbonate by an equal amount incremental dilution method to obtain the veterinary antidiarrhea powder.

[0010] 3. The method for determining the content of the veterinary antidiarrhea powder is to determine the content of berberine hydrochloride and phenyl salicylate by high performance liquid chromatography;

[0011] (1) Determination of berberine hydrochloride content

[0012] Chromatographic conditions: Octadecylsilane bonded silica gel was used as filler, 2-4 g potassium dihydrogen phosphate and 0.5-2 g sodium dodecyl sulfate were dissolved in 1000 mL of a water-acetonitrile mixture with a volume ratio of 1:1 as the mobile phase, and the detection wavelength was 345 nm;

[0013] Preparation of reference solution: Accurately weigh berberine hydrochloride reference substance, dissolve and dilute with mobile phase to prepare reference solution;

[0014] Preparation of test solution: accurately weigh veterinary antidiarrhea powder, add mobile phase, stir to dissolve, centrifuge to obtain supernatant, filter, discard the initial filtrate, and take the subsequent filtrate as the test solution;

[0015] Determination method: Accurately aspirate the reference solution and the test solution respectively, inject into the liquid chromatograph, and record the chromatogram; calculate the content of berberine hydrochloride in the test sample by the peak area according to the external standard method;

[0016] (2) Determination of phenyl salicylate content

[0017] Chromatographic conditions: octadecylsilane bonded silica gel as filler, acetonitrile-water mixture with a volume ratio of 3:7 as mobile phase, detection wavelength of 310 nm;

[0018] Preparation of reference solution: Accurately weigh phenyl salicylate reference, dissolve and dilute with mobile phase to prepare reference solution;

[0019] Preparation of test solution: accurately weigh veterinary antidiarrhea powder, add mobile phase, stir to dissolve, centrifuge to obtain supernatant, filter, discard the initial filtrate, and take the subsequent filtrate as the test solution;

[0020] Determination method: Accurately aspirate the reference solution and the test solution respectively, inject into the liquid chromatograph, and record the chromatogram; calculate the content of phenyl salicylate in the test sample by the peak area according to the external standard method.

[0021] The beneficial effects of the present invention are as follows: the present invention provides an antidiarrhea powder for animals, which has an advanced and reasonable prescription design, can effectively resist bacteria and inflammation, and can quickly stop diarrhea. At the same time, it uses various nutrients contained in yeast to ensure the nutritional absorption of sick animals, quickly restore their digestive function, and can effectively treat intestinal diseases of livestock. Moreover, it can be mixed with feed and fed, and used together with machinery, which is convenient for use in farms and greatly solves the problem of human cost of drug use in farms. This patent also provides a method for determining the content of berberine hydrochloride and phenyl salicylate, the main ingredients in antidiarrhea powder for animals, which has strong specificity, good precision and high accuracy, and can be used for quality control of antidiarrhea powder for animals to ensure the stability of product quality and the safety and effectiveness of veterinary drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is the chromatogram of berberine hydrochloride reference solution.

[0023] Figure 2 This is the chromatogram of the test solution for the determination of berberine hydrochloride content.

[0024] Figure 3 This is the chromatogram of phenyl salicylate reference solution.

[0025] Figure 4 This is the chromatogram of the test solution for the determination of phenyl salicylate content. DETAILED DESCRIPTION

[0026] In order to make the objectives, technical solutions and beneficial effects of the present invention more clear, the preferred embodiments of the present invention are described in detail below.

[0027] Example 1 Preparation of veterinary antidiarrhea powder

[0028] The veterinary antidiarrhea powder is composed of the following components by weight: 20 parts of berberine hydrochloride, 2 parts of phenyl salicylate, 30 parts of montmorillonite, 18 parts of yeast and 30 parts of calcium carbonate.

[0029] Preparation method:

[0030] Mix 2 g of phenyl salicylate and 30 g of montmorillonite, then add 20 g of berberine hydrochloride and 18 g of yeast, mix well, and mix the resulting mixture with 30 g of calcium carbonate by an equal amount incremental dilution method to obtain veterinary antidiarrheal powder.

[0031] Example 2 Determination of the content of veterinary antidiarrhea powder

[0032] The contents of berberine hydrochloride and phenyl salicylate, the main ingredients in veterinary antidiarrhea powder, were determined by HPLC.

[0033] 1. Determination of berberine hydrochloride content

[0034] Chromatographic conditions: The chromatographic column was a C18 column (250 mm × 4.6 mm, 5 μm), 3.4 g of potassium dihydrogen phosphate and 1.7 g of sodium dodecyl sulfate were dissolved in 1000 mL of a water-acetonitrile (volume ratio 1:1) mixture as the mobile phase, the flow rate was 1.0 ml / min, the column temperature was 40 °C, and the detection wavelength was 345 nm.

[0035] Preparation of reference solution: Accurately weigh about 0.03 g of berberine hydrochloride reference substance, dissolve it in mobile phase and dilute to 100 mL as the reference solution.

[0036] Preparation of the test solution: Accurately weigh about 1 g of veterinary antidiarrhea powder, add 100 mL of mobile phase, stir for 10 min, centrifuge to obtain the supernatant, filter with a membrane filter with a pore size of less than 0.5 μm, discard the initial filtrate, and take the subsequent filtrate as the test solution.

[0037] Determination method: Accurately pipette 10 μl of reference solution and test solution respectively, inject into liquid chromatograph, and record the chromatogram; calculate the content of berberine hydrochloride in the test sample by peak area according to the external standard method.

[0038] The chromatogram of the reference solution is shown in Figure 1 The chromatogram of the test solution is shown in Figure 2 .

[0039] System suitability test: Take 10 μL of the reference solution and measure according to the above chromatographic conditions. The theoretical plate number calculated based on the berberine hydrochloride peak is not less than 8000, and the separation between the berberine hydrochloride peak and the adjacent impurity peak is not less than 1.7.

[0040] 2. Determination of phenyl salicylate content

[0041] Chromatographic conditions: The chromatographic column was a C18 column (250 mm × 4.6 mm, 5 μm), with a mixture of acetonitrile and water (volume ratio 3:7) as the mobile phase, a flow rate of 1.0 ml / min, a column temperature of 40 °C, and a detection wavelength of 310 nm.

[0042] Preparation of reference solution: Accurately weigh about 0.05 g of phenyl salicylate reference substance, dissolve it in mobile phase and dilute to 100 mL as the reference solution.

[0043] Preparation of the test solution: Accurately weigh about 5 g of veterinary antidiarrhea powder, add 100 mL of mobile phase, stir for 10 min, centrifuge to obtain the supernatant, filter with a membrane filter with a pore size of less than 0.5 μm, discard the initial filtrate, and take the subsequent filtrate as the test solution.

[0044] Determination method: Accurately pipette 10 μl of reference solution and test solution respectively, inject into liquid chromatograph, record chromatogram; calculate the content of phenyl salicylate in the test sample by peak area according to external standard method.

[0045] The chromatogram of the reference solution is shown in Figure 3 The chromatogram of the test solution is shown in Figure 4 .

[0046] System suitability test: Take 10 μL of the reference solution and measure according to the above chromatographic conditions. The theoretical plate number calculated based on the berberine hydrochloride peak is not less than 10,000, and the separation between the berberine hydrochloride peak and the adjacent impurity peak is not less than 1.5.

[0047] Example 3 Methodological verification of the content determination method

[0048] 1. Exclusivity

[0049] The following two negative samples were prepared: a negative sample lacking berberine hydrochloride and a negative sample lacking phenyl salicylate. The above two negative samples were taken respectively, and negative sample solutions were prepared according to the preparation method of the corresponding test solution in Example 2, and the corresponding chromatographic conditions in Example 2 were used for determination, and the chromatograms were recorded.

[0050] By comparing the chromatograms of the reference solution and the above two negative sample solutions, it was found that the retention time of each component was consistent, and the negative sample solution had no obvious interference with the retention time position of the negative reference solution, proving that under the chromatographic conditions of the present invention, the components can be well separated and the method has strong specificity.

[0051] 2. Linear Relationship

[0052] Accurately weigh 0.3 g of berberine hydrochloride reference substance, place it in a 25 ml volumetric flask, dissolve it with the corresponding mobile phase and make up to volume, as a berberine hydrochloride reference substance stock solution. Accurately measure 0.5, 5, 10, 15, and 25 ml of berberine hydrochloride reference substance stock solution respectively, place it in a 50 ml volumetric flask, dilute it with the corresponding mobile phase and make up to volume, prepare a series of berberine hydrochloride reference substance dilutions, and measure it using the corresponding chromatographic conditions in Example 2, and record the chromatogram. With the injection volume X of the reference substance dilution as the abscissa and the peak area Y as the ordinate, draw a standard curve to obtain a regression equation.

[0053] Accurately weigh 0.5 g of phenyl salicylate reference substance, place it in a 25 ml volumetric flask, dissolve it with the corresponding mobile phase and make up to volume, as a phenyl salicylate reference substance stock solution; accurately measure 0.5, 5, 10, 15, 25 ml of phenyl salicylate reference substance stock solution respectively, place it in a 50 ml volumetric flask, dilute it with the corresponding mobile phase and make up to volume, prepare a series of phenyl salicylate reference substance dilutions, measure it according to the corresponding chromatographic conditions in Example 2, and record the chromatogram. With the injection amount X of the reference substance dilution as the abscissa and the peak area Y as the ordinate, draw a standard curve to obtain a regression equation.

[0054] The results are shown in Table 1. Berberine hydrochloride and phenyl salicylate showed good linear relationships with the peak area in the concentration ranges of 0.12-6.0 and 0.2-10.0 mg / ml, respectively. The correlation coefficient R 2 All are greater than 0.999.

[0055] Table 1 Linear relationship investigation results

[0056] Tested ingredients Regression equation Linear range <![CDATA[R 2 ]]> Berberine Hydrochloride Y=434912X+17744 0.12~6.0mg / ml 0.9995 Phenyl salicylate <![CDATA[Y=5×10 6 X+36806]]> 0.2~10.0mg / ml 0.9998

[0057] 3. Precision

[0058] Accurately pipette 10 μL of the berberine hydrochloride reference solution and phenyl salicylate reference solution prepared in Example 2, perform continuous sampling and measurement 6 times according to the corresponding chromatographic conditions in Example 2, record the chromatogram, and calculate the relative standard deviation (RSD) based on the 6 peak areas of the chromatographic peaks of each active ingredient.

[0059] The results showed that the RSDs of the peak areas of the two active ingredients, berberine hydrochloride and phenyl salicylate, were both less than 1.2%, 0.58% and 0.86% respectively, which proved that the method of the present invention had good precision.

[0060] 4. Sample recovery rate

[0061] Accurately weigh 1 g of veterinary antidiarrhea powder sample with known content into a 100 ml volumetric flask, a total of 6 portions, and accurately add berberine hydrochloride reference solution and phenyl salicylate reference solution according to low, medium and high concentrations, respectively. Prepare the test solution according to the corresponding method in Example 2, and perform sampling and measurement according to the corresponding chromatographic conditions in Example 2, record the chromatogram, calculate the content of the measured component, and calculate the average sample recovery rate of the measured component according to the amount of reference substance added.

[0062] The results are shown in Table 2. The average recoveries of berberine hydrochloride and phenyl salicylate were both greater than 99%, and RSD≤1.2%, which proves that the method of the present invention has high accuracy.

[0063] Table 2 Results of sample recovery investigation

[0064] Tested ingredients Berberine Hydrochloride Phenyl salicylate Average recovery rate (%) 99.45 99.34 RSD(%) 0.43 0.24

[0065] Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that various changes can be made in form and details without departing from the scope defined by the claims of the present invention.

Claims

1. An antidiarrhea powder for animals, characterized in that: The invention is composed of the following components by weight: 1 to 20 parts of berberine hydrochloride, 1 to 5 parts of phenyl salicylate, 10 to 30 parts of montmorillonite, 1 to 30 parts of yeast and 1 to 30 parts of calcium carbonate.

2. The antidiarrhea powder for animals according to claim 1, characterized in that: The invention is composed of the following components by weight: 20 parts of berberine hydrochloride, 2 parts of phenyl salicylate, 30 parts of montmorillonite, 18 parts of yeast and 30 parts of calcium carbonate.

3. The method for preparing the antidiarrhea powder for animals according to claim 1 or 2, characterized in that: The following steps are involved: Phenyl salicylate and montmorillonite are mixed evenly, and then berberine hydrochloride and yeast are added and mixed evenly. The obtained mixture is mixed evenly with calcium carbonate by an equal amount incremental dilution method to obtain veterinary antidiarrheal powder.

4. The method for determining the content of the veterinary antidiarrhea powder according to claim 1 or 2, characterized in that: The contents of berberine hydrochloride and phenyl salicylate were determined by HPLC; (1) Determination of berberine hydrochloride content Chromatographic conditions: Octadecylsilane bonded silica gel was used as filler, 2-4 g potassium dihydrogen phosphate and 0.5-2 g sodium dodecyl sulfate were dissolved in 1000 mL of a water-acetonitrile mixture with a volume ratio of 1:1 as the mobile phase, and the detection wavelength was 345 nm; Preparation of reference solution: Accurately weigh berberine hydrochloride reference substance, dissolve and dilute with mobile phase to prepare reference solution; Preparation of test solution: accurately weigh veterinary antidiarrhea powder, add mobile phase, stir to dissolve, centrifuge to obtain supernatant, filter, discard the initial filtrate, and take the subsequent filtrate as the test solution; Determination method: Accurately aspirate the reference solution and the test solution respectively, inject into the liquid chromatograph, and record the chromatogram; calculate the content of berberine hydrochloride in the test sample by the peak area according to the external standard method; (2) Determination of phenyl salicylate content Chromatographic conditions: octadecylsilane bonded silica gel as filler, acetonitrile-water mixture with a volume ratio of 3:7 as mobile phase, detection wavelength of 310 nm; Preparation of reference solution: Accurately weigh phenyl salicylate reference, dissolve and dilute with mobile phase to prepare reference solution; Preparation of test solution: accurately weigh veterinary antidiarrhea powder, add mobile phase, stir to dissolve, centrifuge to obtain supernatant, filter, discard the initial filtrate, and take the subsequent filtrate as the test solution; Determination method: Accurately aspirate the reference solution and the test solution respectively, inject into the liquid chromatograph, and record the chromatogram; calculate the content of phenyl salicylate in the test sample by the peak area according to the external standard method.

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