Traditional Chinese medicine composition for preventing special aquatic liver and gall diseases
By combining aqueous two-phase extraction and bullfrog bile concentration technology with soybean lecithin and stevioside to form bile acid-phospholipid-stevioside ternary complex micelles, enteric-coated microcapsules are prepared, solving the problems of strong bitterness and poor absorption of components in traditional Chinese medicine, and achieving efficient prevention and treatment of liver and gallbladder diseases in special aquatic frogs.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- FUZHOU PENGXIN ANIMAL HUSBANDRY & AQUATIC PRODUCTS COMPREHENSIVE DEVELOPMENT CO LTD
- Filing Date
- 2026-02-25
- Publication Date
- 2026-05-19
AI Technical Summary
Existing technologies are insufficient to effectively prevent liver and gallbladder diseases in special aquatic frogs. Traditional Chinese medicine powders are bitter and difficult to digest and absorb, resulting in slow and weak efficacy. Traditional bile powders contain impurities and have unstable activity, leading to poor prevention or treatment effects.
By employing aqueous two-phase extraction technology and bullfrog bile compound concentration technology, combined with soybean lecithin and stevioside to form bile acid-phospholipid-stevioside ternary complex micelles, enteric-coated microparticles were prepared. The aqueous two-phase system was used to achieve efficient separation and purification of traditional Chinese medicine components and improve bioavailability. Bile was purified by chitosan flocculation technology to form a stable nanoscale dispersion system.
It significantly improved the bioavailability and pharmacological synergistic effect of traditional Chinese medicine components, increased the feeding rate of frogs and the prevention effect of hepatobiliary diseases, reduced the mortality rate, and achieved precise targeted treatment of hepatobiliary diseases.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of aquatic liver and gallbladder diseases, and more particularly to a traditional Chinese medicine composition for the prevention of liver and gallbladder diseases in specific aquatic animals. Background Technology
[0002] Specialty frog farming (especially the high-density factory farming of bullfrogs) has become an important aquaculture industry. However, in the pursuit of high growth rates and high stocking densities, the frog's hepatobiliary system is under tremendous stress, and hepatobiliary syndrome has become one of the leading non-infectious diseases causing slow growth, low feed conversion rates, decreased immunity, and mass mortality in frogs. Frogs have a relatively high metabolic rate, but their liver and gallbladder functions are simpler compared to mammals. When artificially fed high-energy, high-fat formulated feeds, their liver and gallbladder's fat metabolism and detoxification functions are easily overloaded. Frogs mainly rely on sight to hunt live prey. Even if formulated feeds are nutritionally complete, they cannot completely mimic live prey in terms of shape, movement, and smell, leading to feeding stress and unstable appetite, which in turn causes uneven nutrient intake and fluctuations in liver and gallbladder burden. In aquaculture, high-frequency feeding is often used, and feed is easily dissolved and oxidized in water, producing harmful substances such as aldehydes and ketones, which directly damage liver cells after being ingested. Some traditional Chinese herbs with heat-clearing, detoxifying, and liver-protecting properties (such as Artemisia capillaris, rhubarb, Isatis indigotica, and Gardenia jasminoides) are simply ground into powder and mixed into feed or decocted and sprinkled on the frogs. The herbs have an extremely bitter taste, severely affecting the already picky frogs' appetite, often leading to refusal to eat, making prevention or treatment impossible. The active ingredients in the coarse powder are difficult for frogs to digest and absorb, resulting in slow and weak efficacy. Some feed companies have tried adding animal bile powder, but simply dried bile powder contains a large amount of large-molecule proteins, impurities, and potential pathogens. The concentration of the active ingredient (bile acids) is low and its activity is unstable. It has a strong bitter taste, requiring large amounts of attractants to mask it. As a single ingredient, it fails to combine with a systemic liver-protecting traditional Chinese medicine formula. Purifying animal bile (such as chicken bile) typically involves removing proteins using methods such as ethanol precipitation, acid precipitation, or organic solvent extraction. These methods may leave solvent residues, lead to loss of active ingredients, or be complex processes. Summary of the Invention
[0003] In order to overcome the above-mentioned shortcomings of the prior art, the present invention proposes a traditional Chinese medicine composition for the prevention of liver and gallbladder diseases in special aquatic animals.
[0004] To solve the above-mentioned technical problems, the present invention adopts the following technical solution: a traditional Chinese medicine composition for preventing liver and gallbladder diseases in special aquatic animals, comprising the following steps: Grinding: The traditional Chinese medicine formula is ground into powder and graded to obtain Chinese medicine powder; The traditional Chinese medicine formula includes: 150-160 parts gentian root, 65-75 parts gardenia fruit, 35-45 parts scutellaria root, 130-140 parts plantain, 50-60 parts bupleurum root, 30-40 parts licorice root, and 40-50 parts angelica root. Aqueous two-phase system: At 20-25℃, 16-18% PEG4000, 12-15% K2HPO4 and 3-5% citric acid are added in one step to form two phases; Extraction: Add the Chinese herbal powder to the aqueous two-phase system in a certain proportion, stir at 200-400 r / min for 20-30 min, let stand for 30-40 min, and then perform phase separation and recovery to obtain the aqueous two-phase concentrate. Mixing: The aqueous two-phase concentrate is mixed with the material to prepare microspheres.
[0005] The aqueous two-phase system, with its high water content and mild conditions (room temperature and normal pressure), maximizes the preservation of the bioactivity of heat-sensitive active ingredients (such as glycosides and flavonoids) in traditional Chinese medicine. Utilizing the significant differences in the partition coefficients of target components (gentiopicrin, geniposide, baicalin, etc.) and impurities (proteins, polysaccharides, heavy metal ions) between the two phases, it achieves "one-step separation and purification." Active ingredients are automatically enriched in the upper phase (PEG phase), while proteins, polysaccharides, and harmful heavy metals that affect drug stability and safety precipitate to the lower phase and are removed. This effectively reduces the risk of heavy metal residues in the finished product, meeting the requirements for green aquatic pharmaceuticals. Compared to traditional alcohol extraction and water precipitation methods, the transfer rate of active ingredients is higher.
[0006] Furthermore, the preparation method of the compound concentrated bullfrog bile includes: Healthy bullfrogs were selected, and bile was extracted from them while they were still alive. The bile was stored at 0-4℃, and 0.5-0.7% food-grade chitosan with a degree of deacetylation ≥90% was added to the bile. The mixture was stirred at 25-30℃ for 30-40 minutes to flocculate and precipitate the protein. The mixture was then centrifuged at 8000-10000 r / min for 10-12 minutes to obtain clear bile. The bile was then concentrated 6 times using a 50nm ceramic membrane to obtain a composite concentrated bullfrog bile. The composite concentrated bullfrog bile was mixed with the aqueous two-phase concentrate in a certain proportion to obtain a composite solution. The composite solution was then mixed with other materials to prepare enteric-coated microcapsules. The materials were one or more of feed, medicine, and supplements.
[0007] The amino groups on the chitosan molecular chain are positively charged in weakly acidic to neutral environments, which can efficiently flocculate with negatively charged proteins in bile through charge neutralization and adsorption bridging. It has little effect on bile acids with smaller molecular weights and different charges. Chitosan itself has antibacterial, mucosal repair-promoting and immune-enhancing effects. Its trace residues (if present) are not only harmless, but can also have a synergistic effect on preventing intestinal inflammation in aquatic animals. The creative combination of natural polymer flocculation technology and modern membrane separation technology achieves efficient and clean physical concentration.
[0008] Furthermore, the process includes: adding soybean lecithin and steviol glycosides to the composite solution for dissolution and mixing, wherein soybean lecithin PC≥80% and steviol glycoside Reb-A≥97%, and homogenizing twice under high pressure at 40℃ and 600bar to form bile acid-phospholipid-stevioside ternary composite micelles, ultimately obtaining a modified composite solution, which is the final composite solution product. The bitterness threshold decreased from 0.05% to 0.002% (measured by electronic tongue), and the bullfrog feeding rate increased by 24%.
[0009] The purified aqueous two-phase concentrate of traditional Chinese medicine was mixed with concentrated bullfrog bile at a ratio of 1:5, and then high-purity soybean lecithin and steviol glycosides were added. After high-pressure homogenization, a ternary complex micelle of bile acids, phospholipids, and steviol glycosides was formed. Using soybean lecithin (amphiphilic) as the framework, under the high shear force driven by high-pressure homogenization, it self-assembled with bile acids (amphiphilic) and steviol glycosides (hydrophilic glycoside head + hydrophobic aglycone) to form a "mixed micelle". This is a thermodynamically stable nanoscale dispersion system. Phospholipids and bile acids significantly improve the solubility of lipid-soluble components in the aqueous phase, preventing precipitation. The micelle size (typically tens of nanometers) makes them easily recognized and taken up by the intestinal mucosa. Studies have shown that steviol glycoside Reb-A can affect intestinal epithelial tight junctions, potentially fine-tuning permeability and promoting transmembrane transport of micelles. Steviol glycosides mask bitterness, and their glycoside structure may interact with intestinal carbohydrate receptors, increasing intestinal retention.
[0010] Furthermore, the addition amount of soybean lecithin is 8-10% of the total amount of the compound liquid, and the addition amount of stevioside is 0.3-0.4% of the total amount of the compound liquid.
[0011] Furthermore, the method for preparing enteric-coated microcapsules includes: using 0.8 mm microcrystalline cellulose pellets as seeds, the modified composite solution is sprayed at a rate of 5-7 mL / min into the bottom spray of a centrifugal coating machine, while simultaneously sprinkling in 200-300 mesh corn starch, to obtain 0.6 mm drug-containing pellets with a drug loading of 35%. In this process, acrylic resin IV and chitosan are co-dissolved, with a mass ratio of acrylic resin IV to chitosan of 8:2. 15-18% triethyl citrate is added as a plasticizer, resulting in a coating weight gain of 12-14%. The film-forming temperature is 28-35℃. Then, 0.5% stevioside and 0.3% bullfrog bile attractant are sprayed on the outermost layer, followed by fluidized bed drying at 28℃ for 30 min to obtain enteric-coated microspheres.
[0012] Furthermore, in the re-extraction process, the Chinese herbal powder is added to the system at a material-to-liquid ratio of 1:15. The effective components of the Chinese herbal formula are automatically enriched in the upper phase (PEG phase), while proteins, polysaccharides, and heavy metals precipitate in the lower phase. After phase separation, the upper phase is taken, and the two-phase aqueous concentrate is obtained through the process.
[0013] Furthermore, in the remixing process, the compound concentrated bullfrog bile and the aqueous two-phase concentrate are mixed at a ratio of 1:5.
[0014] Furthermore, the pellet core contains a 5% weight increase in HPMCE5 alcohol-water solution for moisture protection.
[0015] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. The whole formula takes "clearing damp heat in the liver and gallbladder" as its core, while taking into account "soothing the liver and regulating qi" and "nourishing blood and softening the liver". It combines attack and tonification, and uses both cold and warm methods. It is designed for the symptoms of damp heat in the liver and gallbladder and qi stagnation and blood stasis (manifested as hepatomegaly, fatty liver, gallbladder enlargement, etc.) caused by high-fat feed and water quality deterioration in aquatic animals, and prevents them from the perspective of etiology and pathogenesis.
[0016] 2. When combined with traditional Chinese medicine formulas that clear the liver and promote bile secretion, bullfrog bile is not only a traditional "guide drug" for guiding the medicine to its proper channels, but also a precise "biological signaling molecule." It can be better recognized and utilized by the intestinal and enterohepatic circulation systems of the same or closely related species (such as frogs and fish), and may activate relevant receptors and metabolic pathways, thereby greatly enhancing the bioavailability and pharmacological synergistic effect of the entire traditional Chinese medicine formula in the target animal.
[0017] 3. The purified aqueous two-phase concentrate of traditional Chinese medicine was mixed with the compound concentrated bullfrog bile at a ratio of 1:5, and then high-purity soybean lecithin and stevioside were added. The mixture was homogenized under high pressure to form a ternary complex micelle of "bile acid-phospholipid-stevioside".
[0018] 4. This composite micelle mimics the natural form of fat digestion and absorption in mammals (bile acid-phospholipid mixed micelles), which can greatly promote the absorption of hydrophobic drug components in the intestines of aquatic animals and significantly improve bioavailability.
[0019] 5. The coating layer, made of acrylic resin IV (enteric-coating material) and chitosan (bioadhesive and biodegradable), ensures zero drug release in the acidic environment of the stomach, allowing the drug to reach the intestines intact and rapidly disintegrate and release. This avoids damage from gastric acid, protects the acid-labile components of the traditional Chinese medicine, and avoids the influence of gastric emptying, enabling the drug to be released evenly and concentratedly at the main absorption site (intestines), ensuring stable blood drug concentrations and achieving true "intestinal targeted drug delivery," greatly improving the precision and efficiency of treatment.
[0020] 6. This invention achieves multi-functional synergy of masking taste, attracting appetite, and preventing moisture, effectively preventing expensive liver and gallbladder diseases, reducing mortality, and the attracting micro-pellets can be directly mixed with feed for feeding, with low labor intensity and good compliance. Detailed Implementation
[0021] The invention will now be described in detail with reference to the embodiments.
[0022] Example 1
[0023] A traditional Chinese medicine composition for preventing hepatobiliary diseases in special aquatic animals, comprising the following steps: Grinding: The traditional Chinese medicine formula is ground into powder and graded to obtain Chinese medicine powder; The traditional Chinese medicine formula includes: 150 parts gentian root, 65 parts gardenia fruit, 35 parts scutellaria root, 130 parts plantain, 50 parts bupleurum root, 30 parts licorice root, and 40 parts angelica root. Aqueous two-phase system: At 20°C, 16% PEG4000, 12% K2HPO4 and 3% citric acid are added in one step to form two phases; Extraction: The Chinese herbal powder was added to the aqueous two-phase system at a material-to-liquid ratio of 1:15, stirred at 200 r / min for 20 min, and allowed to stand for 30 min. Then, phase separation and recovery were carried out. The effective components of the Chinese herbal formula were automatically enriched in the upper phase, while proteins, polysaccharides, and heavy metals precipitated in the lower phase. After phase separation, the upper phase was taken and processed to obtain the aqueous two-phase concentrate. Mixing: The compound concentrated bullfrog bile was mixed with the above aqueous two-phase concentrate at a ratio of 1:5 to obtain a compound solution. Soybean lecithin and steviol glycosides were added to the compound solution for dissolution and mixing. The amount of soybean lecithin added was 8% of the total amount of the compound solution, and the amount of steviol glycosides added was 0.3% of the total amount of the compound solution. The soybean lecithin (PC) content was ≥80%, and the steviol glycoside (Reb-A) content was ≥97%. The mixture was homogenized twice under high pressure at 40℃ and 600 bar to form bile acid-phospholipid-steviol ternary complex micelles, ultimately obtaining the modified compound solution. 0.8 mm microcrystalline cellulose pellets were used as seeds, wherein the pellet weight gain was [not specified]. A 5% HPMCE5 alcohol-water solution, protected from moisture, was sprayed at a rate of 5 mL / min into the bottom spray of a centrifugal coating machine, while simultaneously sprinkling in 200-mesh corn starch, to obtain 0.6 mm drug-containing pellet cores with a drug loading of 35%. The cores were co-dissolved with acrylic resin IV and chitosan at a mass ratio of 8:2, with 15% triethyl citrate added as a plasticizer, resulting in a 12% weight gain. The film-forming temperature was 28℃. The outermost layer was then sprayed with 0.5% stevioside and 0.3% bullfrog bile attractant, followed by fluidized bed drying at 28℃ for 30 min to obtain enteric-coated microcapsules.
[0024] The preparation method of the compound concentrated bullfrog bile includes: Healthy bullfrogs were used to extract bile from live animals. The bile was stored at 0℃ and 0.5% food-grade chitosan with a degree of deacetylation ≥90% was added to the bile. The mixture was stirred at 25℃ for 30 minutes to flocculate and precipitate the protein. The mixture was then centrifuged at 8000 r / min for 10 minutes to obtain clear bile. The bile was then concentrated 6 times using a 50 nm ceramic membrane to obtain a compound concentrated bullfrog bile.
[0025] Example 2
[0026] A traditional Chinese medicine composition for preventing hepatobiliary diseases in special aquatic animals, comprising the following steps: Grinding: The traditional Chinese medicine formula is ground into powder and graded to obtain Chinese medicine powder; The traditional Chinese medicine formula includes: 155 parts gentian root, 70 parts gardenia fruit, 40 parts scutellaria root, 135 parts plantain, 55 parts bupleurum root, 35 parts licorice root, and 45 parts angelica root. Aqueous two-phase system: At 23°C, 17% PEG4000, 13% K2HPO4 and 4% citric acid are added in one step to form two phases; Extraction: The Chinese herbal powder was added to the aqueous two-phase system at a material-to-liquid ratio of 1:15, stirred at 300 r / min for 25 min, and allowed to stand for 35 min. Then, phase separation and recovery were carried out. The effective components of the Chinese herbal formula were automatically enriched in the upper phase (PEG phase), while proteins, polysaccharides, and heavy metals precipitated in the lower phase. After phase separation, the upper phase was taken and processed to obtain the aqueous two-phase concentrate. Mixing: The compound concentrated bullfrog bile was mixed with the above aqueous two-phase concentrate at a ratio of 1:5 to obtain a compound solution. Soybean lecithin and steviol glycosides were added to the compound solution for dissolution and mixing. The amount of soybean lecithin added was 10% of the total amount of the compound solution, and the amount of steviol glycosides added was 0.3% of the total amount of the compound solution. The soybean lecithin (PC) content was ≥80%, and the steviol glycoside (Reb-A) content was ≥97%. The mixture was homogenized twice at 40℃ and 600 bar to form bile acid-phospholipid-steviol ternary complex micelles, ultimately obtaining a modified compound solution. 0.8 mm microcrystalline cellulose pellets were used as seeds, wherein the pellets increased... A 5% (w / w) HPMCE5 alcohol-water solution was prepared and kept moisture-proof. The modified composite solution was sprayed at a rate of 6 mL / min in the bottom spray of a centrifugal coating machine, while 250-mesh corn starch was sprinkled in simultaneously to obtain 0.6 mm drug-containing pellet cores with a drug loading of 35%. Acrylic resin IV and chitosan were co-dissolved, with a mass ratio of acrylic resin IV to chitosan of 8:2. 17% triethyl citrate was added as a plasticizer, resulting in a 13% weight gain in coating. The film-forming temperature was 30℃. The outermost layer was then sprayed with 0.5% stevioside and 0.3% bullfrog bile attractant, and then dried in a fluidized bed at 28℃ for 30 min to obtain enteric-coated microcapsules.
[0027] The preparation method of the compound concentrated bullfrog bile includes: Healthy bullfrogs were used to extract bile from live animals. The bile was stored at 3°C. 0.6% food-grade chitosan with a degree of deacetylation ≥90% was added to the bile. The mixture was stirred at 27°C for 35 minutes to flocculate and precipitate the protein. The mixture was then centrifuged at 9000 r / min for 11 minutes to obtain clear bile. The bile was then concentrated 6 times using a 50 nm ceramic membrane to obtain a compound concentrated bullfrog bile.
[0028] Example 3
[0029] A traditional Chinese medicine composition for preventing hepatobiliary diseases in special aquatic animals, comprising the following steps: Grinding: The traditional Chinese medicine formula is ground into powder and graded to obtain Chinese medicine powder; The traditional Chinese medicine formula includes: 160 parts gentian root, 75 parts gardenia fruit, 45 parts scutellaria root, 140 parts plantain, 60 parts bupleurum root, 40 parts licorice root, and 50 parts angelica root. Aqueous two-phase system: At 25°C, 18% PEG4000, 15% K2HPO4 and 5% citric acid are added in one step to form two phases; Extraction: The Chinese herbal powder was added to the aqueous two-phase system at a material-to-liquid ratio of 1:15, stirred at 400 r / min for 30 min, and allowed to stand for 40 min. Then, phase separation and recovery were carried out. The effective components of the Chinese herbal formula were automatically enriched in the upper phase (PEG phase), while proteins, polysaccharides, and heavy metals precipitated in the lower phase. After phase separation, the upper phase was taken and processed to obtain the aqueous two-phase concentrate. Mixing: The compound concentrated bullfrog bile was mixed with the above aqueous two-phase concentrate at a ratio of 1:5 to obtain a compound solution. Soybean lecithin and steviol glycosides were added to the compound solution for dissolution and mixing. The amount of soybean lecithin added was 10% of the total amount of the compound solution, and the amount of steviol glycosides added was 0.4% of the total amount of the compound solution. The soybean lecithin (PC) content was ≥80%, and the steviol glycoside (Reb-A) content was ≥97%. The mixture was homogenized twice at 40℃ and 600 bar to form bile acid-phospholipid-steviol ternary complex micelles, ultimately obtaining the modified compound solution. Using 0.8 mm microcrystalline cellulose pellets as seeds, its... In the process, HPMCE5 alcoholic aqueous solution with a 5% increase in core weight was used to prevent moisture. The modified composite solution was sprayed at a rate of 7 mL / min in the bottom spray of a centrifugal coating machine, while 300-mesh corn starch was sprinkled in simultaneously, resulting in 0.6 mm drug-containing cores with a drug loading of 35%. Acrylic resin IV and chitosan were co-dissolved, with a mass ratio of acrylic resin IV to chitosan of 8:2. 18% triethyl citrate was added as a plasticizer, resulting in a 14% increase in coating weight. The film-forming temperature was 35℃. The outermost layer was then sprayed with 0.5% stevioside and 0.3% bullfrog bile attractant, and then dried in a fluidized bed at 28℃ for 30 min to obtain enteric-coated microcapsules.
[0030] The preparation method of the compound concentrated bullfrog bile includes: Healthy bullfrogs were taken, and bile was extracted from them while they were still alive. The bile was stored at 4°C, and 0.7% food-grade chitosan with a degree of deacetylation ≥90% was added to the bile. The mixture was stirred at 30°C for 40 minutes to flocculate and precipitate the protein. The mixture was then centrifuged at 10,000 r / min for 12 minutes to obtain clear bile. The bile was then concentrated 6 times using a 50 nm ceramic membrane to obtain a compound concentrated bullfrog bile.
[0031] Compare with Example 1 Compared with Example 1, Comparative Example 1 did not add compound concentrated bullfrog bile, but was otherwise the same as Example 1.
[0032] Compare with Example 2 Compared with Example 1, Comparative Example 2 did not add a two-phase aqueous system process, but was otherwise the same as Example 1.
[0033] Compare with Example 3 Compared with Example 1, in the process of preparing compound concentrated bullfrog bile, Comparative Example 3 did not use food-grade chitosan as a flocculant, but instead used ethanol, while the rest was the same as in Example 1.
[0034] Compare with Example 4 Compared with Example 1, in the process of preparing the compound concentrated bullfrog bile, Comparative Example 4 did not use food-grade chitosan as a flocculant, but instead used organic solvent extraction to remove protein. The rest was the same as in Example 1.
[0035] Compare with Example 5 Compared with Example 1, in the process of preparing compound concentrated bullfrog bile, Comparative Example 5 did not use food-grade chitosan as a flocculant or a 50nm ceramic membrane for circulation. Instead, it directly used ethanol as a flocculant and then carried out conventional extraction. The rest was the same as in Example 1.
[0036] Compare with Example 6 Compared with Example 1, Comparative Example 6 did not add soybean lecithin and stevioside to the composite solution for dissolution and mixing, and did not form bile acid-phospholipid-stevioside ternary complex micelles. Otherwise, it was the same as Example 1.
[0037] Compare with Example 7 Compared with Example 1, Comparative Example 7 did not use acrylic resin IV-chitosan co-solution, but was otherwise the same as Example 1.
[0038] Traditional Chinese medicine compositions were prepared according to the preparation methods of Examples 1-3 and Comparative Examples 1-7, and experiments were conducted on them, as follows: Frogs with hepatobiliary syndrome were selected, with 50 frogs in each group for experimental comparison, for a total of 11 groups: 10 drug treatment groups and 1 control group. The observation period was 14 days. The 10 drug treatment groups used the traditional Chinese medicine compositions prepared in Examples 1-3 and Control Examples 1-7, respectively. The liver-to-body ratio, ALT / AST, liver fat deposition, and survival rate were tested. The specific data are shown in Table 1. At the beginning of the experiment, data on liver-to-body ratio, ALT / AST, and liver fat deposition were obtained first, and data were obtained again through calculations after the experiment. Liver-to-body weight ratio was calculated based on body weight; ALT / AST ratio was determined using a kit; liver steatosis was stained with Oil Red O; survival rate was calculated statistically.
[0039] As can be seen from Table 1, Examples 1-3 are the optimal solutions.
[0040] The above embodiments merely illustrate several implementation methods of the present invention, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the patent and its protection scope should be determined by the appended claims.
Claims
1. A traditional Chinese medicine composition for preventing liver and gallbladder diseases in special aquatic animals, characterized in that, Includes the following steps: Grinding: The traditional Chinese medicine formula is ground into powder and graded to obtain Chinese medicine powder; The traditional Chinese medicine formula includes: 150-160 parts gentian root, 65-75 parts gardenia fruit, 35-45 parts scutellaria root, 130-140 parts plantain, 50-60 parts bupleurum root, 30-40 parts licorice root, and 40-50 parts angelica root. Aqueous two-phase system: At 20-25℃, 16-18% PEG4000, 12-15% K2HPO4 and 3-5% citric acid are added in one step to form two phases; Extraction: Add the Chinese herbal powder to the aqueous two-phase system in a certain proportion, stir at 200-400 r / min for 20-30 min, let stand for 30-40 min, and then perform phase separation and recovery to obtain the aqueous two-phase concentrate. Mixing: The aqueous two-phase concentrate is mixed with the material to prepare microspheres.
2. The traditional Chinese medicine composition for preventing liver and gallbladder diseases in special aquatic animals as described in claim 1, characterized in that, The mixing process also includes: Healthy bullfrogs were used to extract bile from live animals. The bile was then stored at low temperature. 0.5-0.7% food-grade chitosan with a degree of deacetylation ≥90% was added to the bile. The mixture was stirred at 25-30℃ for 30-40 minutes to flocculate and precipitate proteins. The mixture was then centrifuged at 8000-10000 r / min for 10-12 minutes to obtain clear bile. The bile was then concentrated 6 times using a 50nm ceramic membrane to obtain a composite concentrated bullfrog bile. The composite concentrated bullfrog bile was mixed with the aqueous two-phase concentrate in a certain proportion to obtain a composite solution. The composite solution was then mixed with the materials to prepare enteric-coated microspheres.
3. The traditional Chinese medicine composition for preventing liver and gallbladder diseases in special aquatic animals as described in claim 2, characterized in that, Also includes: Soybean lecithin and stevioside were added to the composite solution for dissolution and mixing, wherein soybean lecithin PC≥80% and stevioside Reb-A≥97%. The mixture was homogenized twice under high pressure at 40℃ and 600bar to form bile acid-phospholipid-stevioside ternary composite micelles, and the modified composite solution was finally obtained. This modified composite solution is the final composite solution product.
4. The traditional Chinese medicine composition for preventing liver and gallbladder diseases in special aquatic animals as described in claim 3, characterized in that, Also includes: The amount of soybean lecithin added is 8-10% of the total amount of the compound solution, and the amount of stevioside added is 0.3-0.4% of the total amount of the compound solution.
5. The traditional Chinese medicine composition for preventing liver and gallbladder diseases in special aquatic animals as described in claim 1, characterized in that, It also includes a method for preparing enteric-coated microcapsules: using 0.8 mm microcrystalline cellulose pellets as seeds, the above-mentioned modified composite solution is sprayed at a rate of 5-7 mL / min in the bottom spray of a centrifugal coating machine, while simultaneously sprinkling in 200-300 mesh corn starch, to obtain 0.6 mm drug-containing pellets with a drug loading of 35%. In this process, acrylic resin IV and chitosan are co-dissolved, with a mass ratio of acrylic resin IV to chitosan of 8:
2. 15-18% triethyl citrate is added as a plasticizer, resulting in a coating weight gain of 12-14%. The film-forming temperature is 28-35℃. Then, 0.5% stevioside and 0.3% bullfrog bile attractant are sprayed on the outermost layer, followed by fluidized bed drying at 28℃ for 30 min to obtain enteric-coated microspheres.
6. The traditional Chinese medicine composition for preventing liver and gallbladder diseases in special aquatic animals as described in claim 2, characterized in that, In the re-extraction process, the Chinese herbal powder is added to the system at a material-to-liquid ratio of 1:
15. The effective components of the Chinese herbal formula are automatically enriched in the upper phase (PEG phase), while proteins, polysaccharides, and heavy metals precipitate in the lower phase. After phase separation, the upper phase is taken, and the two-phase aqueous concentrate is obtained through the process.
7. A traditional Chinese medicine composition for preventing liver and gallbladder diseases in special aquatic animals as described in any one of claims 2-4, characterized in that, In the remixing process, the compound concentrated bullfrog bile and the aqueous two-phase concentrate are mixed at a ratio of 1:
5.
8. The traditional Chinese medicine composition for preventing liver and gallbladder diseases in special aquatic animals as described in claim 5, characterized in that, in, HPMCE5 alcoholic aqueous solution with 5% weight gain of pellet core, moisture-proof.