Veterinary compound astragalus polysaccharide injection and production process thereof

Abstract

The invention discloses a veterinary compound astragalus polysaccharide injection and a production process thereof, and belongs to the technical field of veterinary medicine preparations, and the production process comprises the following steps: dissolving astragalus polysaccharide, and compounding with composite nanoparticles to obtain astragalus polysaccharide loaded powder; adding a surfactant and absolute ethyl alcohol into liquid paraffin, adding the dispersion liquid into a reaction kettle while stirring, and then stirring to obtain the compound astragalus polysaccharide injection; according to the production process disclosed by the invention, cyclodextrin is prepared into a carrier for adsorbing and packaging the astragalus polysaccharide, a core-shell structure is formed by the cyclodextrin carrier and the polylactic acid by utilizing the adsorption capacity of the cyclodextrin carrier, and the polylactic acid coated with nano-selenium is loaded in the cyclodextrin carrier, so that agglomeration of nano-selenium in the injection is prevented, and the dispersibility of the astragalus polysaccharide is also improved; the composite nanoparticles are prepared into the emulsion, so that the dispersity of the composite nanoparticles is improved, the injection in an emulsion form is more easily absorbed by animals, the nano-selenium and the astragalus polysaccharide generate a synergistic effect, and the resistance of the animals is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of veterinary pharmaceutical preparations, and specifically relates to a veterinary compound astragalus polysaccharide injection and a production process thereof. Background Art

[0002] Astragalus polysaccharide is the most important natural active ingredient in Astragalus. It is a class of macromolecular compounds that plays a decisive role in the pharmacological effects of Astragalus. Astragalus polysaccharide can improve the body's immune function, enhance cell physiological metabolism, and increase macrophage activity. It is an ideal immunopotentiator. Astragalus polysaccharide injection is currently a relatively ideal, safe and effective preparation. It has a significant effect on improving animal immunity, can effectively reduce the outbreak and transmission of epidemic diseases in the breeding industry, and provide protection for animal husbandry.

[0003] Patent CN102198152A discloses an injection containing astragalus polysaccharide iron for pigs and a preparation method thereof. Guided by nanotechnology, astragalus polysaccharide and trivalent iron salt are used as the main raw materials. The iron core of ferric hydroxide is synthesized, and then complexed with an alkalinized astragalus polysaccharide ligand, refined, formulated and sterilized to obtain an astragalus polysaccharide iron injection, which can effectively enhance the body's immunity and improve the survival rate of piglets. However, this scheme uses astragalus polysaccharide to disperse iron ions in the injection to improve the utilization rate of iron ions by piglets, and the synergistic effect with astragalus polysaccharide is low.

[0004] Patent CN105456185A discloses a compound astragalus polysaccharide injection and a preparation method thereof. The solution viscosity is increased by adding water-soluble polymer material PVPk30 and polyethylene glycol, the drug diffusion rate is reduced, and the action time is prolonged. By selecting two natural polysaccharides from different sources, astragalus polysaccharide and α-mannan peptide, the immune enhancement effect is effectively exerted. However, the use of polyethylene glycol to increase the solution viscosity in this scheme easily leads to a decrease in the solubility of the polysaccharide, easy precipitation, and difficulty in absorption, resulting in poor injection effect and being unfavorable for storage and transportation.

[0005] A stable dispersion of astragalus polysaccharide is needed, which is convenient for storage and transportation, has good injection effect, and has good absorption effect of astragalus polysaccharide, so as to enhance the immunity of animals and promote the growth of animals. Summary of the invention

[0006] One of the purposes of the present invention is to solve how to improve the stable dispersibility of astragalus polysaccharide in astragalus polysaccharide injection, and to provide a veterinary compound astragalus polysaccharide injection and its production process.

[0007] The second purpose of the present invention is to solve how to improve the absorption effect of compound astragalus polysaccharide injection in animals and the immunity of animals, and to provide a veterinary compound astragalus polysaccharide injection and its production process.

[0008] The purpose of the present invention can be achieved through the following technical solutions: A production process of a veterinary compound astragalus polysaccharide injection comprises the following steps: Step 1: Dissolve astragalus polysaccharide in deionized water in a reaction kettle, add the composite nanoparticles into the reaction kettle, stir for 40-48 hours, collect the precipitate by centrifugation, wash the precipitate with ethanol, and vacuum dry it at 40-50° C. for 10-12 hours to obtain astragalus polysaccharide loaded powder.

[0009] Step 2: Add surfactant and anhydrous ethanol to liquid paraffin in a reaction kettle, ultrasonically disperse the astragalus polysaccharide-loaded powder in deionized water to obtain a dispersion, then add the dispersion to the reaction kettle at 35-40° C. while stirring, and then stir for 18-20 hours to obtain the compound astragalus polysaccharide injection.

[0010] In the dispersion, the dosage ratio of astragalus polysaccharide to deionized water is 1.8-2g:200-300mL.

[0011] The surfactant is one of Span60, Span80, Tween60 and Tween80 or a mixture of two of them.

[0012] Furthermore, in step 1, the dosage ratio of astragalus polysaccharide, deionized water and composite nanoparticles is 6-8g:20-25mL:1.8-2g.

[0013] Furthermore, in step 2, the dosage ratio of the surfactant, anhydrous ethanol, liquid paraffin and dispersion is 30-35 g: 10-12 mL: 20-25 g: 20-30 g.

[0014] Further, the composite nanoparticles are prepared by the following steps: The polylactic acid / nano-selenium microspheres were dispersed in deionized water to prepare a 5 g / L polylactic acid dispersion. The cyclodextrin carrier and the polyvinyl alcohol surfactant were added to the deionized water in a reactor. The polylactic acid dispersion was added to the reactor and stirred for 10-12 hours. The precipitate was collected after ultrafiltration to obtain composite nanoparticles.

[0015] Furthermore, the usage ratio of the cyclodextrin carrier, the polyvinyl alcohol surfactant, the deionized water and the polylactic acid dispersion is 1.5-2 g: 0.01-0.02 g: 80-100 mL: 150-200 mL.

[0016] Further, the cyclodextrin carrier is prepared by the following steps: Potassium hydroxide and β-cyclodextrin are dissolved in deionized water, anhydrous ethanol is added, ultrasonicated and heated to 60-70°C for reaction for 1-1.5 hours, polyethylene glycol is added for reaction for 10-12 hours, the precipitate is collected by centrifugation, washed and vacuum dried at 40-50°C for 10-12 hours to obtain a cyclodextrin carrier.

[0017] Furthermore, the usage ratio of potassium hydroxide, β-cyclodextrin, deionized water, anhydrous ethanol and polyethylene glycol is 1-1.5 g: 3-4 g: 10-15 mL: 60-80 mL: 1-1.5 g.

[0018] Further, the polylactic acid / nano-selenium microspheres are prepared by the following steps: Dissolve polylactic acid in acetone in a reactor, add nano-selenium into the reactor, stir for 20-30 minutes, and then ultrasonicate in an ice bath for 15-20 minutes. Then add it into a polyvinyl alcohol solution with a concentration of 5 g / L and stir at 1000-1200 rpm for 10-12 hours. After standing overnight, collect the precipitate by centrifugation, wash the precipitate with deionized water, and then freeze-dry to obtain polylactic acid / nano-selenium microspheres.

[0019] Furthermore, the usage ratio of polylactic acid, acetone, nano-selenium and polyvinyl alcohol solution is 4-5g:100-150mL:0.5-0.7g:0.8-1L.

[0020] The beneficial effects of the present invention are as follows: (1) The production process of the present invention prepares cyclodextrin as a carrier to adsorb and encapsulate astragalus polysaccharide, and utilizes the adsorption capacity of the cyclodextrin carrier to form a core-shell structure with polylactic acid, and loads the polylactic acid coated with nano-selenium in the cyclodextrin carrier, thereby preventing the aggregation of nano-selenium in the injection solution and improving the dispersibility of astragalus polysaccharide. By preparing the composite nanoparticles into an emulsion, the dispersibility of the composite nanoparticles is improved, and the injection solution in the form of an emulsion is more easily absorbed by animals, so that the nano-selenium and astragalus polysaccharide produce a synergistic effect and improve the resistance of animals. (2) The veterinary compound astragalus polysaccharide injection prepared by the present invention is an emulsion with high dispersibility and good permeability. The composite nanoparticles in the emulsion are prepared by compounding astragalus polysaccharide with nano-selenium to improve the resistance of animals, coating the nano-selenium with polylactic acid to reduce the agglomeration of nano-selenium, and compounding the cyclodextrin carrier with polylactic acid to form a core-shell structure through the adsorption capacity of the cyclodextrin carrier. The astragalus polysaccharide is then immersed and coated in the cyclodextrin carrier to improve the dispersibility of the astragalus polysaccharide. The cyclodextrin and polylactic acid are easily decomposed by animals and are non-toxic to animals, and have a good effect on improving the resistance of animals. DETAILED DESCRIPTION

[0021] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0022] Example 1: A production process of a veterinary compound astragalus polysaccharide injection, comprising the following steps: S1. Dissolve 4 g of polylactic acid in 100 mL of acetone in a reactor, add 0.5 g of nano-selenium into the reactor, stir for 20 min, ultrasonicate in an ice bath for 15 min, then add into 0.8 L of 5 g / L polyvinyl alcohol solution and stir at 1000 rpm for 10 h. After standing overnight, collect the precipitate by centrifugation, wash the precipitate with deionized water and freeze-dry to obtain polylactic acid / nano-selenium microspheres.

[0023] S2. Dissolve 1 g potassium hydroxide and 3 g β-cyclodextrin in 10 mL deionized water, add 60 mL anhydrous ethanol, sonicate and heat to 60 °C for 1 h, then add 1 g polyethylene glycol for 10 h, collect the precipitate by centrifugation, wash with anhydrous ethanol and dry under vacuum at 40 °C for 10 h to obtain a cyclodextrin carrier.

[0024] S3. Disperse the polylactic acid / nano-selenium microspheres in deionized water to prepare 5 g / L polylactic acid dispersion. Add 1.5 g of cyclodextrin carrier and 0.01 g of polyvinyl alcohol surfactant into 80 mL of deionized water in a reactor. Add 150 mL of polylactic acid dispersion into the reactor and stir to react for 10 h. Collect the precipitate after ultrafiltration to obtain composite nanoparticles.

[0025] S4. Dissolve 6 g of astragalus polysaccharide in 205 mL of deionized water in a reactor, add 1.8 g of composite nanoparticles into the reactor, stir for 40 h, collect the precipitate by centrifugation, wash the precipitate with ethanol, and vacuum dry at 40° C. for 10 h to obtain astragalus polysaccharide loaded powder.

[0026] S5. Add 30 g of Span60 and 10 mL of anhydrous ethanol to 20 g of liquid paraffin in a reactor, ultrasonically disperse 1.8 g of astragalus polysaccharide-loaded powder in 200 mL of deionized water to obtain a dispersion, then add 20 g of the dispersion into the reactor at 35° C. while stirring, and then stir for 18 h to obtain the compound astragalus polysaccharide injection.

[0027] Embodiment 2: A production process of a veterinary compound astragalus polysaccharide injection, comprising the following steps: S1. Dissolve 4.5 g of polylactic acid in 125 mL of acetone in a reactor, add 0.6 g of nano-selenium into the reactor, stir for 25 min, ultrasonicate in an ice bath for 17 min, then add into 0.9 L of 5 g / L polyvinyl alcohol solution and stir at 1100 rpm for 11 h. After standing overnight, collect the precipitate by centrifugation, wash the precipitate with deionized water and freeze-dry to obtain polylactic acid / nano-selenium microspheres.

[0028] S2. Dissolve 1.25 g potassium hydroxide and 3.5 g β-cyclodextrin in 12.5 mL deionized water, add 70 mL anhydrous ethanol, sonicate and heat to 65 ° C for 1.25 h, then add 1.25 g polyethylene glycol for 11 h, collect the precipitate by centrifugation, wash with anhydrous ethanol and dry under vacuum at 45 ° C for 11 h to obtain a cyclodextrin carrier.

[0029] S3. Disperse the polylactic acid / nano-selenium microspheres in deionized water to prepare 5 g / L polylactic acid dispersion. Add 1.75 g of cyclodextrin carrier and 0.015 g of polyvinyl alcohol surfactant into 90 mL of deionized water in a reactor. Add 175 mL of polylactic acid dispersion into the reactor and stir to react for 11 h. Collect the precipitate after ultrafiltration to obtain composite nanoparticles.

[0030] S4. Dissolve 7 g of astragalus polysaccharide in 22.5 mL of deionized water in a reactor, add 1.9 g of composite nanoparticles into the reactor, stir for 44 h, collect the precipitate by centrifugation, wash the precipitate with ethanol, and dry it in vacuum at 45°C for 11 h to obtain astragalus polysaccharide loaded powder.

[0031] S5. Add 10 g Span60, 22.5 g Span80 and 11 mL anhydrous ethanol to 22.5 g liquid paraffin in a reactor, ultrasonically disperse 1.9 g astragalus polysaccharide-loaded powder in 250 mL deionized water to obtain a dispersion, then add 25 g of the dispersion into the reactor at 37°C while stirring, and then stir for 19 hours to obtain the compound astragalus polysaccharide injection.

[0032] Embodiment 3: A production process of a veterinary compound astragalus polysaccharide injection, comprising the following steps: S1. Dissolve 5 g of polylactic acid in 150 mL of acetone in a reactor, add 0.7 g of nano-selenium into the reactor, stir for 30 min, ultrasonicate in an ice bath for 20 min, then add into 1 L of 5 g / L polyvinyl alcohol solution and stir at 1200 rpm for 12 h. After standing overnight, collect the precipitate by centrifugation, wash the precipitate with deionized water and freeze-dry to obtain polylactic acid / nano-selenium microspheres.

[0033] S2. Dissolve 1.5 g potassium hydroxide and 4 g β-cyclodextrin in 15 mL deionized water, add 80 mL anhydrous ethanol, sonicate and heat to 70 °C for 1.5 h, then add 1.5 g polyethylene glycol for 12 h, collect the precipitate by centrifugation, wash with anhydrous ethanol and dry under vacuum at 50 °C for 12 h to obtain a cyclodextrin carrier.

[0034] Cyclodextrin as an organic ligand is combined with potassium ions through coordination bonds to produce a cyclodextrin carrier, which has adsorption and encapsulation capabilities, is safe and non-toxic, and can be degraded by animals.

[0035] S3. Disperse the polylactic acid / nano-selenium microspheres in deionized water to prepare 5 g / L polylactic acid dispersion. Add 2 g of cyclodextrin carrier and 0.02 g of polyvinyl alcohol surfactant to 100 mL of deionized water in a reactor. Add 200 mL of polylactic acid dispersion to the reactor, stir and react for 12 hours. Collect the precipitate after ultrafiltration to obtain composite nanoparticles.

[0036] Under the action of polyvinyl alcohol surfactant, polylactic acid / nano-selenium microspheres and cyclodextrin carriers are adsorbed on the polylactic acid / nano-selenium microspheres to form composite nanoparticles with polylactic acid / nano-selenium microspheres as cores and cyclodextrin carriers as shells, so that the nano-selenium is separated and the aggregation of nano-selenium is reduced. Polylactic acid and cyclodextrin are safe and non-toxic and can be degraded and metabolized by animals. Nano-selenium can provide animals with selenium, promote their growth, and improve their resistance.

[0037] S4. Dissolve 8 g of astragalus polysaccharide in 25 mL of deionized water in a reactor, add 2 g of composite nanoparticles into the reactor, stir for 48 h, collect the precipitate by centrifugation, wash the precipitate with ethanol, and vacuum dry at 50° C. for 12 h to obtain astragalus polysaccharide loaded powder.

[0038] Through the impregnation method, the adsorption capacity of the cyclodextrin carrier in the composite nanoparticles is utilized to adsorb and encapsulate astragalus polysaccharide on the surface of the cyclodextrin carrier, thereby reducing the precipitation of astragalus polysaccharide caused by long-term standing. The combined effect of astragalus polysaccharide and nano-selenium can improve the resistance of animals.

[0039] S5. Add 10 g of Tween80, 25 g of Tween60 and 12 mL of anhydrous ethanol to 25 g of liquid paraffin in a reactor, ultrasonically disperse 2 g of astragalus polysaccharide-loaded powder in 300 mL of deionized water to obtain a dispersion, then add 30 g of the dispersion into the reactor at 40° C. while stirring, and then stir for 20 hours to obtain the compound astragalus polysaccharide injection.

[0040] After the astragalus polysaccharide-loaded powder is dispersed in water, it forms a nanoemulsion with liquid paraffin under the action of a surfactant. The nanoemulsion has high stability and is easy to transport and store. It has high dispersibility and good permeability after injection, and works synergistically with nano-selenium to have a good effect on improving the resistance of animals.

[0041] Comparative Example 1: The difference from Example 1 is that nano-selenium is used to replace polylactic acid / nano-selenium microspheres to prepare compound astragalus polysaccharide injection.

[0042] Comparative Example 2: The difference from Example 1 is that the composite nanoparticles are replaced by a cyclodextrin carrier to prepare a compound astragalus polysaccharide injection.

[0043] Comparative Example 3: The difference from Example 1 is that the composite nanoparticles are replaced with a cyclodextrin carrier to prepare an astragalus polysaccharide cyclodextrin carrier, and the astragalus polysaccharide loaded powder is replaced with the above-mentioned astragalus polysaccharide cyclodextrin carrier and polylactic acid nano-selenium microspheres.

[0044] The performance of the compound astragalus polysaccharide injection prepared in Example 1 to Example 3 and Comparative Example 1 to Comparative Example 3 was tested. 120 40-day-old piglets in the nursery stage were randomly selected and divided into 6 groups, with 20 pigs in each group. 0.2 mL of the compound astragalus polysaccharide injection was injected intramuscularly per kilogram of body weight once a week for 3 weeks. In the fourth week, 5 mL of blood was collected from each pig in the six groups, of which 2.5 mL was whole blood and 2.5 mL was used to prepare serum to determine the superoxide dismutase (SOD) activity, the immunoglobulin of the pig was determined by immunoturbidimetry, and the red blood cell count was determined by the test tube dilution method, and the results were averaged.

[0045] The results are shown in Table 1: project Example 1 Example 2 Example 3 Comparative Example 1 Comparative Example 2 Comparative Example 3 Immunoglobulin (g / L) 136.35 139.72 141.49 114.21 120.56 126.17 <![CDATA[Red blood cell count (10 9 / L)]]> 11.89 12.13 12.41 9.17 10.22 10.76 SOD(U / mL) 168.24 173.26 176.05 145.38 152.16 159.54 As shown in Table 1, the compound astragalus polysaccharide injection prepared by the present invention can significantly improve the immunoglobulin and red blood cell count of piglets, and has high SOD activity, indicating that the compound astragalus polysaccharide injection prepared by the present invention has a good immunity improvement effect on piglets.

[0046] In Comparative Example 1, since nano-selenium is directly compounded with the cyclodextrin carrier, the nano-selenium is deposited in the pores of the cyclodextrin carrier, which reduces the loading amount of the cyclodextrin carrier on astragalus polysaccharide, and the improvement of the immune ability of piglets is poor.

[0047] Since no polylactic acid / nano-selenium microspheres were added to Comparative Example 2, there was no synergistic effect of nano-selenium, and the improvement of the immune ability of piglets was worse than that of Examples 1 to 3 and Comparative Example 3.

[0048] In Comparative Example 3, since astragalus polysaccharide is loaded on a cyclodextrin carrier and then added to the oil phase with polylactic acid / nano-selenium microspheres to prepare an emulsion, the dispersibility of nano-selenium and astragalus polysaccharide is different, the synergistic effect is small, and the improvement of the immune ability of piglets is worse than that of Examples 1 to 3.

[0049] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A production process of a veterinary compound astragalus polysaccharide injection, characterized in that: The steps include: Step 1: dissolve astragalus polysaccharide in deionized water in a reaction kettle, add the composite nanoparticles into the reaction kettle, stir for 40-48 hours, collect the precipitate by centrifugation, wash the precipitate with ethanol, and then vacuum dry at 40-50° C. for 10-12 hours to obtain astragalus polysaccharide loaded powder; Step 2: Add a surfactant and anhydrous ethanol to liquid paraffin in a reaction kettle, ultrasonically disperse the astragalus polysaccharide-loaded powder in deionized water to obtain a dispersion, then add the dispersion to the reaction kettle at 35-40° C. while stirring, and then stir for 18-20 hours to obtain a compound astragalus polysaccharide injection; The composite nanoparticles are prepared by the following steps: The polylactic acid / nano-selenium microspheres are dispersed in deionized water to prepare a 5 g / L polylactic acid dispersion, a cyclodextrin carrier and a polyvinyl alcohol surfactant are added to the deionized water in a reaction kettle, the polylactic acid dispersion is added to the reaction kettle, the reaction is stirred for 10-12 hours, and the precipitate is collected after ultrafiltration to obtain composite nanoparticles; In the dispersion, the dosage ratio of astragalus polysaccharide to deionized water is 1.8-2 g: 200-300 mL; The surfactant is one of Span60, Span80, Tween60 and Tween80, or a mixture of two of them.

2. The production process of a veterinary compound astragalus polysaccharide injection according to claim 1, characterized in that: The dosage ratio of astragalus polysaccharide, deionized water and composite nanoparticles in step 1 is 6-8g:20-25mL:1.8-2g.

3. The production process of a veterinary compound astragalus polysaccharide injection according to claim 1, characterized in that: The dosage ratio of the surfactant, anhydrous ethanol, liquid paraffin and dispersion in step 2 is 30-35g: 10-12mL: 20-25g: 20-30g.

4. The production process of a veterinary compound astragalus polysaccharide injection according to claim 1, characterized in that: The dosage ratio of the cyclodextrin carrier, the polyvinyl alcohol surfactant, the deionized water and the polylactic acid dispersion is 1.5-2g: 0.01-0.02g: 80-100mL: 150-200mL.

5. The production process of a veterinary compound astragalus polysaccharide injection according to claim 4, characterized in that: The cyclodextrin carrier is prepared by the following steps: Potassium hydroxide and β-cyclodextrin are dissolved in deionized water, anhydrous ethanol is added, ultrasonicated and heated to 60-70°C for reaction for 1-1.5 hours, polyethylene glycol is added for reaction for 10-12 hours, the precipitate is collected by centrifugation, washed and vacuum dried at 40-50°C for 10-12 hours to obtain a cyclodextrin carrier.

6. The production process of a veterinary compound astragalus polysaccharide injection according to claim 5, characterized in that: The dosage ratio of potassium hydroxide, beta-cyclodextrin, deionized water, anhydrous ethanol and polyethylene glycol is 1-1.5 g: 3-4 g: 10-15 mL: 60-80 mL: 1-1.5 g.

7. The production process of a veterinary compound astragalus polysaccharide injection according to claim 6, characterized in that: The polylactic acid / nano-selenium microspheres are prepared by the following steps: Dissolve polylactic acid in acetone in a reactor, add nano-selenium into the reactor, stir for 20-30 minutes, and then ultrasonicate in an ice bath for 15-20 minutes. Then add it into a polyvinyl alcohol solution with a concentration of 5 g / L and stir at 1000-1200 rpm for 10-12 hours. After standing overnight, collect the precipitate by centrifugation, wash the precipitate with deionized water, and then freeze-dry to obtain polylactic acid / nano-selenium microspheres.

8. The production process of a veterinary compound astragalus polysaccharide injection according to claim 7, characterized in that: The dosage ratio of the polylactic acid, acetone, nano-selenium and polyvinyl alcohol solution is 4-5g: 100-150mL: 0.5-0.7g: 0.8-1L.

9. A veterinary compound astragalus polysaccharide injection, characterized in that: The invention is prepared by the production process described in any one of claims 1 to 8.