Preparation process of a compound traditional Chinese medicine granule preparation for veterinary use

Abstract

This application relates to the technical field of important veterinary preparations and discloses a preparation process for a compound traditional Chinese medicine granule preparation for veterinary use, including the following steps: (1) extracting dandelion, indigo leaf, isatis root, honeysuckle, licorice, patchouli, and gypsum with water to obtain an aqueous extract; adding scutellaria and phellodendron to the residue after water extraction and extracting with alcohol to obtain an alcohol extract; combining the extracts and concentrating to obtain a clear extract; (2) first mixing hydroxypropyl-β-cyclodextrin and sulfobutyl ether-β-cyclodextrin to obtain a mixed cyclodextrin solution, adding glycyrrhizic acid for pre-inclusion to form a glycyrrhizic acid-cyclodextrin pre-inclusion complex; then adding the obtained clear extract to continue inclusion to form an inclusion complex suspension; (3) mixing the inclusion complex suspension with pharmaceutical excipients, granulating and drying to obtain the final product. This application can improve the inclusion rate of baicalin and berberine and improve storage stability.

Description

Technical Field

[0001] This application relates to the technical field of important veterinary preparations, and in particular to a preparation process for a compound traditional Chinese medicine granule preparation for veterinary use. Background Technology

[0002] Currently, for bacterial inflammations in livestock and poultry (such as respiratory tract infections, enteritis, and urinary tract infections caused by Escherichia coli, Salmonella, and Streptococcus), most commercially available traditional Chinese medicine preparations use raw materials such as dandelion, Isatis leaf, and Isatis root directly mixed and pulverized into powders, or use the traditional single-method water decoction to extract the active ingredients and then granulate them. The core steps of the traditional preparation process are: raw material cleaning → cutting into sections → single water decoction 1-2 times → combining filtrates → atmospheric pressure concentration → spray drying granulation. The products are mostly marketed in the form of powders or coarse granules.

[0003] Compound preparations contain both water-soluble components (polysaccharides, alkaloid salts, chlorogenic acid, etc.) and fat-soluble components (baicalin, berberine, etc.). Traditional single-solvent extraction cannot simultaneously extract both types of components, resulting in a low extraction rate of active ingredients. Preparations obtained through direct pulverization or simple extraction lack a protective mechanism for active ingredients, making them susceptible to decomposition and oxidation due to light, temperature, and humidity. After 6 months of storage, the loss rate of active ingredients reaches over 25%, resulting in a short shelf life and causing inconvenience for large-scale livestock farming and storage. Summary of the Invention

[0004] In order to achieve efficient simultaneous extraction of water-soluble and fat-soluble components in compound preparations and improve the solubility and stability of baicalin and berberine-like fat-soluble components, this application provides a preparation process for veterinary compound traditional Chinese medicine granules.

[0005] The technical solution adopted in this application is as follows: A preparation process for a compound traditional Chinese medicine granule formulation for veterinary use includes the following steps; (1) Dandelion, Isatis leaf, Isatis root, honeysuckle, licorice, patchouli and gypsum were extracted with water to obtain an aqueous extract; Scutellaria baicalensis and Phellodendron bark were added to the residue after water extraction and extracted with alcohol to obtain an alcohol extract; the extracts were combined and concentrated to obtain a clear extract. (2) First, hydroxypropyl-β-cyclodextrin and sulfobutyl ether-β-cyclodextrin are mixed to obtain a mixed cyclodextrin solution. Glycyrrhizic acid is added for pre-inclusion to form a glycyrrhizic acid-cyclodextrin pre-inclusion complex. Then, the obtained extract is added to continue the inclusion to form an inclusion complex suspension. (3) Mix the inclusion complex suspension with pharmaceutical excipients, granulate and dry to obtain the final product.

[0006] By employing the above-mentioned technical solution, the medicinal raw materials are divided into two groups for sequential extraction. The first group (dandelion, indigo leaf, isatis root, honeysuckle, licorice, patchouli, and gypsum) undergoes water extraction first to enrich water-soluble components such as polysaccharides, alkaloid salts, and chlorogenic acid. The second group (Scutellaria baicalensis and Phellodendron amurense) undergoes alcohol extraction on the residue after water extraction. Utilizing the cell wall swelling effect of the medicinal materials, ethanol can more easily penetrate into the interior of the medicinal materials, efficiently extracting fat-soluble components such as baicalin (flavonoids) and berberine (alkaloids). A ternary system is constructed by combining hydroxypropyl-β-cyclodextrin and sulfobutyl ether-β-cyclodextrin, and introducing glycyrrhizic acid. Hydroxypropyl-β-cyclodextrin has a high affinity for neutral flavonoids (baicalin) and encapsulates baicalin through hydrophobic interactions. Sulfobutyl ether-β-cyclodextrin carries a negative charge and exhibits both hydrophobic inclusion and electrostatic attraction to positively charged quaternary ammonium alkaloids (berberine). Glycyrrhizic acid, acting as a "bridging molecule," has its hydrophobic end (glycyrrhetinic acid) encapsulated by hydroxypropyl-β-cyclodextrin. The hydrophilic sugar chain extends outward and interacts with the phenolic hydroxyl groups of baicalin through hydrogen bonds, forming a ternary complex of "cyclodextrin-glycyrrhizic acid-baicalin." Therefore, the inclusion component in this ternary system can significantly improve the inclusion rate of baicalin and berberine.

[0007] Optionally, the raw materials in step (1) include, by weight, the following: 25-35 parts of dandelion, 20-28 parts of indigo leaf, 20-28 parts of isatis root, 10-15 parts of honeysuckle, 5-10 parts of licorice, 5-8 parts of patchouli, 3-6 parts of gypsum, 8-12 parts of scutellaria, and 8-12 parts of phellodendron bark.

[0008] Optionally, in step (2), the molar ratio of hydroxypropyl-β-cyclodextrin, sulfobutyl ether-β-cyclodextrin and glycyrrhizic acid is (7-9):(1-2):(0.3-0.8).

[0009] By employing the above technical solution, hydroxypropyl-β-cyclodextrin exhibits good water solubility and strong inclusion capacity, playing a dominant role in the formulation and ensuring efficient inclusion of baicalin. The negatively charged sulfonyl side chain of sulfobutyl ether-β-cyclodextrin primarily includes berberine, exhibiting both electrostatic attraction and hydrophobic inclusion effects; a suitable dosage is sufficient for efficient binding of the alkaloid. Glycyrrhizic acid, acting as a "bridging molecule," has its hydrophobic end included by hydroxypropyl-β-cyclodextrin, while the hydrophilic sugar chain binds to baicalin via hydrogen bonds, forming a ternary complex. This complex effectively bridges the alkaloid without excessively occupying the cyclodextrin cavity.

[0010] Optionally, the inclusion process in step (2) adopts segmented pH control: first adjust the pH to 4.0-5.0 for inclusion for 20-40 min, and then adjust the pH to 6.0-6.5 for inclusion for 50-70 min.

[0011] By employing the above technical solution, in the acidic stage, alkaloids such as berberine are protonated and carry a positive charge, which strongly attracts the sulfonic acid groups (carrying a negative charge) of sulfobutyl ether-β-cyclodextrin, promoting the entry of alkaloids into the cavity of sulfobutyl ether-β-cyclodextrin or their binding to its surface. Subsequently, under near-neutral pH conditions, baicalin exists in a molecular state with enhanced hydrophobicity, facilitating its entry into the cavity of hydroxypropyl-β-cyclodextrin; simultaneously, the carboxyl group of glycyrrhizic acid is deprotonated, enhancing the hydrogen bond interaction with the phenolic hydroxyl group of baicalin, forming a stable ternary complex.

[0012] Optionally, in step (2), after pH adjustment, the temperature is first raised to 48-52℃ and kept at that temperature for 50-70 minutes, and then lowered to 32-35℃ and kept at that temperature for 20-40 minutes.

[0013] By employing the above technical solutions, increasing the temperature promotes molecular thermal motion, accelerates the recognition and binding of host and guest molecules, improves inclusion kinetic efficiency, and allows already recognized molecular pairs to more fully penetrate deep into the cavity. Slow cooling promotes the orderly arrangement and crystallization of inclusion compounds, forming inclusion particles with uniform particle size and stable structure, improving particle flowability and enhancing long-term stability.

[0014] Optionally, in step (2), after mixing hydroxypropyl-β-cyclodextrin and sulfobutyl ether-β-cyclodextrin, the mixture is heated to 45-50°C, and then glycyrrhizic acid is added, and the mixture is kept warm and stirred.

[0015] By adopting the above technical solution, the mixed cyclodextrin solution is first preheated to a specified temperature, and then glycyrrhizic acid is added to ensure that pre-inclusion occurs immediately at the optimal temperature after the addition of glycyrrhizic acid, forming a glycyrrhizic acid-cyclodextrin pre-inclusion complex. This step allows the hydrophobic end (glycyrrhetinic acid) of glycyrrhizic acid to be encapsulated by the cavity of hydroxypropyl-β-cyclodextrin, and the hydrophilic sugar chain extends outward to form a "molecular brush" structure on the surface of cyclodextrin, providing hydrogen bond interaction sites for subsequent binding of baicalin.

[0016] Optionally, during the heating process, the heating rate is 1℃ / min, and during the cooling process, the cooling rate is 0.5℃ / min.

[0017] Optionally, in step (3), the pharmaceutical excipients include maltodextrin and steviol glycosides.

[0018] Optionally, in step (1), the raw material is first dried and then pulverized through a 40-60 mesh sieve.

[0019] By adopting the above technical solution,

[0020] In summary, this application includes at least one of the following beneficial effects: 1. In the technical solution of this application, hydroxypropyl-β-cyclodextrin and sulfobutyl ether-β-cyclodextrin are combined, and glycyrrhizic acid is introduced to construct a ternary system. Hydroxypropyl-β-cyclodextrin has a high affinity for neutral flavonoids (baicalin) and encapsulates baicalin through hydrophobic interactions. Sulfobutyl ether-β-cyclodextrin carries a negative charge and exhibits both hydrophobic encapsulation and electrostatic attraction to positively charged quaternary ammonium alkaloids (berberine). Glycyrrhizic acid, as a "bridging molecule," has its hydrophobic end (glycyrrhetinic acid) encapsulated by hydroxypropyl-β-cyclodextrin, and its hydrophilic sugar chain extends outward, interacting with the phenolic hydroxyl groups of baicalin through hydrogen bonds to form a "cyclodextrin-glycyrrhizic acid-baicalin" ternary complex. Therefore, the inclusion component of the ternary system in this invention can greatly improve the inclusion rate of baicalin and berberine. Detailed Implementation

[0021] The present application will be further described in detail below with reference to the embodiments.

[0022] All medicinal materials used in this application were dried before use, then pulverized and passed through a 40-mesh sieve for later use. Example 1

[0023] This embodiment discloses a preparation process for a compound traditional Chinese medicine granule formulation for veterinary use, including the following steps: (1) Mix 25 parts of dandelion, 28 parts of indigo leaf, 20 parts of isatis root, 10 parts of honeysuckle, 5 parts of licorice, 5 parts of agastache, and 6 parts of gypsum, add 10 times the amount of purified water, heat to 90℃, and extract at a constant temperature for 1.5h. Filter to obtain water extract and residue. (2) Add 8 parts of Scutellaria baicalensis and 8 parts of Phellodendron chinense to the obtained residue, then add 10 times the amount of 70% ethanol, heat to 65℃, extract at constant temperature for 1.5 hours, filter to obtain alcohol extract, mix the water extract and alcohol extract obtained in step (1), and concentrate under reduced pressure to a clear extract with a relative density of 1.15-1.20. (3) Mix 0.7 mol hydroxypropyl-β-cyclodextrin with 0.1 mol sulfobutyl ether-β-cyclodextrin, prepare a 40 wt% mixture with water, stir to dissolve, and obtain a cyclodextrin solution. Then heat the cyclodextrin solution to 45°C, add 0.3 mol glycyrrhizic acid, mix evenly, and obtain a pre-inclusion complex solution. (4) Gradually add the obtained clear extract to the pre-inclusion complex solution, adjust the pH to 4.5 with 10% hydrochloric acid, stir for 30 min, then adjust the pH to 6.5 with 5% sodium hydroxide, stir for 60 min; then raise the temperature to 50℃ at 1℃ / min and keep it warm, then lower the temperature to 35℃ at 0.5℃ / min and keep it warm for 30 min to obtain the inclusion complex suspension; (5) Take the inclusion complex suspension, add maltodextrin (the ratio of solid content of the suspension to the mass of maltodextrin is 1:0.4) and steviol glycosides (0.2%), stir until completely dissolved, and granulate using a boiling granulation machine: inlet air temperature 82℃, outlet air temperature 52℃, spray speed 10mL / min, dry to a moisture content ≤5%, and granulate through a 20-mesh sieve to obtain the final product. Example 2

[0024] This embodiment discloses a preparation process for a compound traditional Chinese medicine granule formulation for veterinary use, including the following steps: (1) Mix 35 parts of dandelion, 20 parts of indigo leaf, 28 parts of isatis root, 15 parts of honeysuckle, 10 parts of licorice, 8 parts of agastache and 3 parts of gypsum, add 10 times the amount of purified water, heat to 90℃, and extract at a constant temperature for 1.5h. Filter to obtain water extract and residue. (2) Add 12 parts of Scutellaria baicalensis and 12 parts of Phellodendron chinense to the obtained residue, then add 10 times the amount of 70% ethanol, heat to 65℃, extract at constant temperature for 1.5 hours, filter to obtain alcohol extract, mix the water extract and alcohol extract obtained in step (1), and concentrate under reduced pressure to a clear extract with a relative density of 1.15-1.20. (3) Mix 0.7 mol hydroxypropyl-β-cyclodextrin with 0.1 mol sulfobutyl ether-β-cyclodextrin, prepare a 40 wt% mixture with water, stir to dissolve, and obtain a cyclodextrin solution. Then heat the cyclodextrin solution to 45°C, add 0.3 mol glycyrrhizic acid, mix evenly, and obtain a pre-inclusion complex solution. (4) Gradually add the obtained clear extract to the pre-inclusion complex solution, adjust the pH to 4.5 with 10% hydrochloric acid, stir for 30 min, then adjust the pH to 6.5 with 5% sodium hydroxide, stir for 60 min; then raise the temperature to 50℃ at 1℃ / min and keep it warm, then lower the temperature to 35℃ at 0.5℃ / min and keep it warm for 30 min to obtain the inclusion complex suspension; (5) Take the inclusion complex suspension, add maltodextrin (the ratio of solid content of the suspension to the mass of maltodextrin is 1:0.4) and steviol glycosides (0.2%), stir until completely dissolved, and granulate using a boiling granulation machine: inlet air temperature 82℃, outlet air temperature 52℃, spray speed 10mL / min, dry to a moisture content ≤5%, and granulate through a 20-mesh sieve to obtain the final product. Example 3

[0025] This embodiment discloses a preparation process for a compound traditional Chinese medicine granule formulation for veterinary use, including the following steps: (1) Mix 25 parts of dandelion, 28 parts of indigo leaf, 15 parts of isatis root, 10 parts of honeysuckle, 5 parts of licorice, 5 parts of agastache, and 6 parts of gypsum, add 10 times the amount of purified water, heat to 90℃, and extract at a constant temperature for 1.5h. Filter to obtain water extract and residue. (2) Add 8 parts of Scutellaria baicalensis and 8 parts of Phellodendron chinense to the obtained residue, then add 10 times the amount of 70% ethanol, heat to 65℃, extract at constant temperature for 1.5 hours, filter to obtain alcohol extract, mix the water extract and alcohol extract obtained in step (1), and concentrate under reduced pressure to a clear extract with a relative density of 1.15-1.20. (3) Mix 0.9 mol hydroxypropyl-β-cyclodextrin with 0.2 mol sulfobutyl ether-β-cyclodextrin, prepare a 40 wt% mixture with water, stir to dissolve, and obtain a cyclodextrin solution. Then heat the cyclodextrin solution to 45°C, add 0.6 mol glycyrrhizic acid, mix evenly, and obtain a pre-inclusion complex solution. (4) Gradually add the obtained clear extract to the pre-inclusion complex solution, adjust the pH to 4.5 with 10% hydrochloric acid, stir for 30 min, then adjust the pH to 6.5 with 5% sodium hydroxide, stir for 60 min; then raise the temperature to 50℃ at 1℃ / min and keep it warm, then lower the temperature to 35℃ at 0.5℃ / min and keep it warm for 30 min to obtain the inclusion complex suspension; (5) Take the inclusion complex suspension, add maltodextrin (the ratio of solid content of the suspension to the mass of maltodextrin is 1:0.4) and steviol glycosides (0.2%), stir until completely dissolved, and granulate using a boiling granulation machine: inlet air temperature 82℃, outlet air temperature 52℃, spray speed 10mL / min, dry to a moisture content ≤5%, and granulate through a 20-mesh sieve to obtain the final product.

[0026] Comparative Example 1 The process steps of this comparative example and Example 1 are the same, except that glycyrrhizic acid was not added in step (3).

[0027] Comparative Example 2 The process steps of this comparative example and Example 1 are the same. The difference is that in steps (3) and (4), the stepwise inclusion process is not used. Instead, glycyrrhizic acid and the extract are directly added to the cyclodextrin solution.

[0028] Comparative Example 3 The process steps of this comparative example and Example 1 are the same, except that the pH is not adjusted in step (4).

[0029] Comparative Example 4 The process steps of this comparative example and Example 1 are the same, except that there is no heating and cooling step in step (4).

[0030] Performance testing The inclusion rates of baicalin and berberine in the formulation particles obtained in Examples 1-3 and Comparative Examples 1-4 were tested. The formulation particles were placed in an environment of 40°C and 70% humidity for 3 months for accelerated testing to test the retention rates of baicalin and berberine (retention rate = inclusion rate after accelerated testing / original inclusion rate * 100%).

[0031] Table 1. Test results of Examples 1-3 and Comparative Examples 1-4 Baicalin inclusion rate / % Baicalin retention rate after accelerated testing / % Berberine inclusion rate / % Berberine retention rate after accelerated testing / % Example 1 96.3 90.1 94.3 91.6 Example 2 96.5 89.8 94.6 91.8 Example 3 96.8 91.3 95.5 92.3 Comparative Example 1 91.1 75.6 88.6 75.2 Comparative Example 2 93.6 83.3 90.3 78.3 Comparative Example 3 92.8 81.4 91.3 80.3 Comparative Example 4 91.3 80.6 90.9 81.6 Comparative data from the examples and comparative cases show that the inclusion complexes containing glycyrrhizic acid exhibit higher inclusion rates for baicalin and berberine, and demonstrate greater stability in accelerated testing, resulting in better retention of the active ingredients. Furthermore, in this application, glycyrrhizic acid is added stepwise to the cyclodextrin solution before being added to the extract, avoiding competitive inhibition and better leveraging the "bridging" effect of glycyrrhizic acid.

[0032] The above are all preferred embodiments of this application, and are not intended to limit the scope of protection of this application. Therefore, all equivalent changes made in accordance with the structure, shape and principle of this application should be covered within the scope of protection of this application.

Claims

1. A preparation process for a compound traditional Chinese medicine granule formulation for veterinary use, characterized in that: Includes the following steps; (1) Dandelion, Isatis leaf, Isatis root, honeysuckle, licorice, patchouli and gypsum were extracted with water to obtain an aqueous extract; Scutellaria baicalensis and Phellodendron chinense were added to the residue after water extraction, and alcohol extraction was carried out to obtain an alcohol extract; the extracts were combined and concentrated to obtain a clear extract. (2) First, hydroxypropyl-β-cyclodextrin and sulfobutyl ether-β-cyclodextrin are mixed to obtain a mixed cyclodextrin solution. Glycyrrhizic acid is added for pre-inclusion to form a glycyrrhizic acid-cyclodextrin pre-inclusion complex. Then, the obtained extract is added to continue the inclusion to form an inclusion complex suspension. (3) Mix the inclusion complex suspension with pharmaceutical excipients, granulate and dry to obtain the final product.

2. The preparation process of a veterinary compound traditional Chinese medicine granule preparation according to claim 1, characterized in that: The raw materials in step (1) include, by weight, 25-35 parts of dandelion, 20-28 parts of indigo leaf, 20-28 parts of isatis root, 10-15 parts of honeysuckle, 5-10 parts of licorice, 5-8 parts of patchouli, 3-6 parts of gypsum, 8-12 parts of scutellaria, and 8-12 parts of phellodendron bark.

3. The preparation process of a veterinary compound traditional Chinese medicine granule preparation according to claim 2, characterized in that: In step (2), the molar ratio of hydroxypropyl-β-cyclodextrin, sulfobutyl ether-β-cyclodextrin and glycyrrhizic acid is (7-9):(1-2):(0.3-0.8).

4. The preparation process of a veterinary compound traditional Chinese medicine granule preparation according to claim 3, characterized in that: The inclusion process described in step (2) adopts segmented pH control: first adjust the pH to 4.0-5.0 for inclusion for 20-40 min, and then adjust the pH to 6.0-6.5 for inclusion for 50-70 min.

5. The preparation process of a veterinary compound traditional Chinese medicine granule formulation according to claim 4, characterized in that: In step (2), after pH adjustment, the temperature is first raised to 48-52℃ and kept for 50-70 minutes, and then lowered to 32-35℃ and kept for 20-40 minutes.

6. The preparation process of a veterinary compound traditional Chinese medicine granule preparation according to claim 1, characterized in that: In step (2), hydroxypropyl-β-cyclodextrin and sulfobutyl ether-β-cyclodextrin are mixed, heated to 45-50°C, and then glycyrrhizic acid is added. The mixture is kept warm and stirred.

7. The preparation process of a veterinary compound traditional Chinese medicine granule preparation according to claim 5, characterized in that: During the heating process, the heating rate is 1℃ / min, and during the cooling process, the cooling rate is 0.51℃ / min.

8. The preparation process of a veterinary compound traditional Chinese medicine granule preparation according to claim 7, characterized in that: In step (3), the pharmaceutical excipients include maltodextrin and steviol glycosides.

9. The preparation process of a veterinary compound traditional Chinese medicine granule preparation according to claim 8, characterized in that: In step (1), the raw material is first dried and then pulverized through a 40-60 mesh sieve.