Preparation method of Xihuang oral liquid for treating chicken damp-heat diarrhea

Abstract

The invention belongs to the technical field of veterinary drugs, and provides a preparation method of Xuhuang oral liquid for treating chicken damp-heat diarrhea, which comprises the following steps: S1, extracting baicalein: purifying by using a macroporous resin column, and purifying the baicalein from scutellaria baicalensis; s2, coating chlorogenic acid: extracting chlorogenic acid from honeysuckle flowers, and coating the chlorogenic acid with cyclodextrin to obtain cyclodextrin-coated chlorogenic acid; s3, traditional Chinese medicine fermentation: mixing radix curcumae, fructus chebulae, fructus gardeniae and radix et rhizoma rhei, and then adding nutrient substances, citric acid and compound bacteria for fermentation to obtain fermentation liquor; s4, purification and sterilization: mixing the baicalein, the cyclodextrin coated chlorogenic acid and the fermentation liquor which are respectively obtained in the steps S1, S2 and S3, purifying, sterilizing and sub-packaging to obtain the Xuhuang oral liquid. The Xuhuang oral liquid disclosed by the invention is high in active ingredient content, the product quality is improved, the taking dosage is reduced, the effect of treating the damp-heat diarrhea of chickens is remarkable, and the Xuhuang oral liquid has market value.

Description

Technical Field

[0001] The invention belongs to the technical field of veterinary drugs, and particularly relates to a preparation method of Yuhuang oral liquid for treating damp-heat diarrhea in chickens. Background Art

[0002] With the rapid development of intensive breeding industry, damp-heat diarrhea in chickens has become an important disease affecting poultry health and breeding efficiency. The disease is usually caused by multiple factors, including bacteria, viruses and environmental stress. The clinical manifestations are lethargy, loss of appetite or bloody stools and diarrhea in chickens.

[0003] At present, the treatment of damp-heat diarrhea in chickens mainly relies on antibiotics, but the long-term abuse of antibiotics has led to drug resistance and drug residue problems, posing a serious threat to food safety and public health. Therefore, the development of new, efficient and safe treatment options has become an urgent need in the field of animal husbandry and veterinary drug research. Traditional Chinese medicine, with its unique treatment concept and fewer side effects, provides a new idea for the treatment of damp-heat diarrhea in chickens, but the extraction and administration methods of traditional Chinese medicine have certain limitations, such as low extraction efficiency, unstable active ingredients, and inconvenient administration.

[0004] Chinese patent CN 104224968 A discloses a Chinese veterinary oral liquid for treating diarrhea in livestock and poultry and its preparation method, which comprises 5-55 parts of Andrographis paniculata, 5-50 parts of Sophora flavescens, 5-40 parts of Coptis chinensis, 0.01-1 parts of potassium sorbate, and 0.05-10 parts of Tween 80 by weight; each component of the Chinese veterinary oral liquid of this invention adopts natural Chinese medicine raw materials, which has simple configuration, wide source of medicine, low cost, no drug residue, small toxic and side effects, and greatly improves the safety of livestock products. However, the natural Chinese medicine raw materials of this invention adopt simple water extraction or alcohol extraction, and the extraction rate of its active ingredients is low, and it is necessary to optimize the extraction process to increase the content of active ingredients, thereby improving the therapeutic effect.

[0005] Therefore, there is an urgent need for a method for preparing Yuhuang oral liquid for treating chicken damp-heat diarrhea, which has high active ingredients and significant therapeutic effect. Summary of the invention

[0006] In view of the existing technical problems, the purpose of the present invention is to provide a method for preparing Yuhuang oral liquid for treating damp-heat diarrhea in chickens. The Yuhuang oral liquid of the present invention has high active ingredients, improves product quality, and thus has a significant effect in treating damp-heat diarrhea in chickens, providing a safe, efficient, and non-toxic new veterinary drug option for the breeding industry.

[0007] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0008] The present invention provides a method for preparing Yuhuang oral liquid for treating chicken damp-heat diarrhea, comprising the following steps:

[0009] S1. Extracting baicalein: purifying baicalein from Scutellaria baicalensis using a macroporous resin column;

[0010] S2. Coated chlorogenic acid: extracting chlorogenic acid from honeysuckle and coating it with cyclodextrin to obtain cyclodextrin-coated chlorogenic acid;

[0011] S3. Chinese medicine fermentation: Mix turmeric, kezi, gardenia, and rhubarb, add nutrients, citric acid, and compound bacteria for fermentation to obtain a fermentation liquid;

[0012] S4. Purification and sterilization: The baicalein, cyclodextrin-coated chlorogenic acid and fermentation liquid obtained in the above steps S1, S2 and S3 are mixed, purified, sterilized and packaged to obtain Yuhuang oral liquid.

[0013] The reaction mechanism and effects of the present invention are as follows:

[0014] 1. The present invention extracts Chinese medicinal ingredients in different ways according to the characteristics of different Chinese medicinal materials, thereby increasing the active ingredients of Yuhuang oral liquid and further improving the treatment rate of chicken damp-heat diarrhea.

[0015] 2. Baicalein, as an active ingredient extracted from Scutellaria baicalensis, has multiple pharmacological effects in the treatment of damp-heat diarrhea in chickens. Its safety and minor side effects are highly favored in veterinary applications. First, baicalein can inhibit a variety of pathogenic bacteria including Shigella dysenteriae, Salmonella typhi and Pseudomonas aeruginosa, providing antibacterial effects for the treatment of damp-heat diarrhea in chickens. It also has antiviral and anti-inflammatory activities, which help to reduce intestinal damage or infection caused by viruses and inflammation; secondly, the antipyretic and diuretic effects of baicalein help to relieve fever symptoms and promote the excretion of toxins in the body; in addition, its antioxidant properties help to protect cells from oxidative stress damage and enhance the body's resistance. However, traditional extraction methods have strict equipment requirements, high costs, low yields, difficult operations, and serious waste liquid pollution.

[0016] The invention uses a self-enzymatic hydrolysis method and further selects a specific macroporous resin column for purification to obtain baicalein. This extraction method not only has little environmental pollution but also has a high conversion rate, and is conducive to extracting high-purity baicalein from Scutellaria baicalensis.

[0017] 3. Chlorogenic acid, the active ingredient in honeysuckle, has significant pharmacological effects in the treatment of damp-heat diarrhea in chickens. First, chlorogenic acid can not only effectively inhibit a variety of pathogenic bacteria including hemolytic streptococci and Staphylococcus aureus, but also has anti-inflammatory and antiviral activity; secondly, the hemostatic effect of chlorogenic acid has a positive effect on the treatment of bloody diarrhea; in addition, chlorogenic acid also has the effect of protecting the liver and promoting bile secretion, which is beneficial to the health of the liver and gallbladder. Compared with chemically synthesized antibiotics, chlorogenic acid is not easy to develop drug resistance and is suitable as a preventive and therapeutic ingredient added to feed for a long time. However, chlorogenic acid may undergo decomposition or isomerization during high-temperature sterilization, affecting its stability and biological activity.

[0018] The present invention uses a water reflux extraction process to extract chlorogenic acid, increases the extraction rate by adjusting process parameters, and then uses cyclodextrin in a specific mass ratio to coat the chlorogenic acid to improve its heat resistance, and further adds surfactants and stabilizers to increase the dispersibility of cyclodextrin-coated chlorogenic acid in water without reducing their coating ability, thereby improving the stability of chlorogenic acid.

[0019] 4. The present invention uses fermentation to extract active ingredients from Curcuma, Fructus Caryophylli, Fructus Gardeniae, and Rhubarb. The reaction conditions are mild, and the effective ingredients of Chinese medicine can be protected from destruction to the maximum extent, so that the content of effective ingredients is increased, the efficacy of the medicine is improved, and the dosage is reduced. The applicant controls the type and proportion of the composite bacteria to destroy the cell wall of the Chinese medicine, so that the effective ingredients are more easily released, thereby improving the extraction rate of the Chinese medicine.

[0020] In some embodiments, the specific steps of step S1 are as follows:

[0021] (1) crushing Scutellaria baicalensis into 50-80 mesh Scutellaria baicalensis powder, soaking in deionized water 16-20 times the mass of the Scutellaria baicalensis powder, heating to 40-60° C., hydrolyzing for 10-14 hours, filtering to obtain a filter residue;

[0022] (2) adding the filter residue obtained in step (1) to ethanol with an amount of 28-32 times the mass of the filter residue, soaking for 5-8 hours, filtering, and distilling under reduced pressure to obtain the filtrate to obtain Scutellaria baicalensis extract;

[0023] (3) Purifying the scutellaria baicalensis extract obtained in step (2) using a macroporous resin column, and drying to obtain baicalin.

[0024] In some embodiments, the macroporous resin column in step (3) is a DM101 macroporous resin column, and the eluent is 60-90 wt % ethanol.

[0025] In some embodiments, the volume ratio of the scutellaria baicalensis extract to the macroporous resin in the macroporous resin column in step (3) is 1:(2-5).

[0026] In some embodiments, the specific steps of step S2 are as follows:

[0027] ① Crush the honeysuckle into 20-50 mesh honeysuckle powder, add it to the reactor, then add deionized water 10-40 times the mass of the honeysuckle powder, soak for 30-60 minutes, reflux extraction 2-4 times, combine the extracts, filter, and concentrate to obtain chlorogenic acid extract;

[0028] ② Dissolve cyclodextrin in deionized water to form a 16-20 g / L cyclodextrin solution, add the chlorogenic acid extract obtained in step ①, and stir while adding to obtain an inclusion complex;

[0029] ③ Add the surfactant and stabilizer to the inclusion compound obtained in step ②, heat to 40-60° C., stir for 12-24 hours, filter, and dry to obtain cyclodextrin-coated chlorogenic acid.

[0030] In some embodiments, the temperature of the reflux extraction in step ① is 60-100° C., and the time of each reflux extraction is 30-90 min.

[0031] In some embodiments, the mass ratio of the chlorogenic acid extract to the cyclodextrin solution in step ② is 1:(10-40).

[0032] In some embodiments, the surfactant and stabilizer in step ③ are 4-8% and 2-4% of the mass of cyclodextrin respectively.

[0033] Specifically, the surfactant is poloxamer 188; and the stabilizer is polyvinyl pyrrolidone.

[0034] Preferably, the cyclodextrin is β-cyclodextrin.

[0035] In some implementations, the specific steps of step S3 are as follows:

[0036] In parts by weight, 10-20 parts of turmeric, 5-15 parts of kohlrabi, 5-10 parts of gardenia and 5-10 parts of rhubarb are mixed and crushed, sterilized, and loaded into a fermentation tank, and a carbon source, a nitrogen source, citric acid and composite bacteria are added, and deionized water is added to adjust the water content to 40-60%, and the mixture is sealed and fermented at 30-37°C for 6-8 days, and then the same amount of composite bacteria and deionized water are added, and the mixture is soaked and extracted at 24-28°C for 26-30 hours, during which the mixture is stirred 2-3 times, centrifuged, filtered, and fermented to obtain a fermentation liquid.

[0037] In some embodiments, the composite bacteria in step S3 is Bacillus subtilis 10 9-12 CFU / g, Bacillus amyloliquefaciens 10 9-12 CFU / g and lactic acid bacteria 10 10-11 CFU / g.

[0038] Preferably, the Bacillus subtilis 10 9-12 CFU / g, Bacillus amyloliquefaciens 10 9-12 CFU / g and lactic acid bacteria 10 10 -11 The mass ratio of CFU / g among the three is 1:(1-1.5):(1-2).

[0039] More preferably, the lactic acid bacteria is Lactobacillus plantarum.

[0040] In some embodiments, the mass ratio of honeysuckle, scutellaria baicalensis and curcuma is 1:(3-4):(4-6).

[0041] Compared with the prior art, the present invention has the following beneficial effects:

[0042] 1. The Yuhuang oral liquid of the present invention not only solves the problems of low extraction efficiency and unstable active ingredients of traditional Chinese medicine, but also improves the application value of the product through the integration and innovation of modern pharmaceutical technology, and provides a new type of veterinary drug for the breeding industry that is safe, efficient and has no toxic side effects.

[0043] 2. The present invention further selects a specific macroporous resin column to purify baicalein by self-enzymatic hydrolysis, which not only has little environmental pollution but also has a high conversion rate, which is conducive to extracting high-purity baicalein from Scutellaria baicalensis. Baicalein can not only inhibit a variety of pathogens, but also has antiviral and anti-inflammatory activities, and also has antioxidant properties, which helps to protect cells from oxidative stress damage; secondly, the antipyretic and diuretic effects of baicalein help to relieve fever symptoms and promote the excretion of toxins in the body.

[0044] 3. Chlorogenic acid, an active ingredient in honeysuckle, has a significant pharmacological effect in treating damp-heat diarrhea in chickens. The present invention uses a water reflux extraction process to extract chlorogenic acid, and then uses a specific ratio of cyclodextrin, surfactant and stabilizer to improve its heat resistance, dispersibility and stability.

[0045] 4. The present invention extracts active ingredients from Curcuma, Fructus Caryophylli, Fructus Gardeniae and Rhubarb by fermentation. By controlling the types and proportions of the composite bacteria, the content of the active ingredients is increased, the extraction rate of the traditional Chinese medicine is improved, and the efficacy of the medicine is improved, thereby reducing the dosage. DETAILED DESCRIPTION

[0046] The present invention will be described below in conjunction with specific embodiments. It should be noted that the following embodiments are examples of the present invention and are only used to illustrate the present invention, rather than to limit the present invention. Other combinations and various modifications within the concept of the present invention may be performed without departing from the spirit or scope of the present invention.

[0047] According to the ratios of the raw materials and the preparation methods specified in the following examples and comparative examples, Yuhuang oral liquid was prepared.

[0048] In order to facilitate those skilled in the art to implement the present invention, some raw material manufacturers of the embodiments and comparative examples are described as follows:

[0049] Polyvinyl pyrrolidone: purchased from Shanxi Jinyang Pharmaceutical Excipients Co., Ltd., model CP2020;

[0050] Bacillus subtilis: purchased from Wuhan Lanabai Pharmaceutical Chemical Co., Ltd., 10 11 CFU / g; Bacillus amyloliquefaciens: purchased from Weifang Ruichen Biotechnology Co., Ltd., 10 12 CFU / g; Lactobacillus plantarum: purchased from Zhongke Jiayi Nutritional Medicine (Shandong) Microecology Research Institute Co., Ltd., 10 11 CFU / g;

[0051] Other raw materials are not specially specified and can be purchased from the market.

[0052] Example 1

[0053] A method for preparing Yuhuang oral liquid for treating chicken damp-heat diarrhea comprises the following steps:

[0054] S1. Extraction of baicalin:

[0055] (1) 10.5 g of Scutellaria baicalensis was crushed into 75 mesh Scutellaria baicalensis powder, soaked in 189 g of deionized water, heated to 50° C., hydrolyzed for 12 h, and filtered to obtain a residue;

[0056] (2) adding the filter residue obtained in step (1) to ethanol with an amount 30 times the mass of the filter residue, soaking for 6.5 hours, filtering, and distilling under reduced pressure to obtain the filtrate to obtain Scutellaria baicalensis extract;

[0057] (3) Purifying the scutellaria baicalensis extract obtained in step (2) by using a macroporous resin column, spray drying, controlling the air inlet temperature at 140° C., the air outlet temperature at 60° C., and the needle-passing operation frequency at 6 seconds / time, to obtain baicalein; wherein the macroporous resin column is a DM101 macroporous resin column, the eluent is 75wt% ethanol, and the volume of the macroporous resin is 3.5 times the volume of the scutellaria baicalensis extract;

[0058] S2. Encapsulated chlorogenic acid:

[0059] ① Crush 3g of honeysuckle into 40-mesh honeysuckle powder, add it to the reactor, then add deionized water 25 times the mass of the honeysuckle powder, soak for 45 minutes, reflux extraction 3 times, the reflux extraction temperature is 80°C, and the reflux extraction time is 60 minutes each time. After the extracts are combined, they are filtered and concentrated to obtain chlorogenic acid extract;

[0060] ② Dissolve 18.5g of β-cyclodextrin in 1000ml of deionized water to form a β-cyclodextrin solution for later use; add the chlorogenic acid extract obtained in step ① to the reactor, and then add a β-cyclodextrin solution with a mass of 25 times that of the chlorogenic acid extract, stirring while adding, and continue stirring for 10 hours after the addition is completed to obtain an inclusion compound;

[0061] ③ Add poloxamer 188 (6% of the mass of β-cyclodextrin used) and polyvinyl pyrrolidone (3% of the mass of β-cyclodextrin used) to the inclusion compound obtained in step ②, heat to 50°C, stir for 20h, filter, and dry at 90°C for 7h to obtain cyclodextrin-coated chlorogenic acid;

[0062] S3. Traditional Chinese medicine fermentation:

[0063] 15g of Curcuma, 10g of Codonopsis pilosula, 7.5g of Gardenia jasminoides, and 7.5g of Rhubarb were mixed and crushed into 80 mesh powder, sterilized at 135°C for 8s, put into a fermentation tank, added with 24g of glucose, 1.2g of urea, 0.3g of citric acid, and 32g of composite bacteria, and deionized water was added to adjust the water content to 50%, and sealed and fermented at 35°C for 7 days, and then the same amount of composite bacteria and deionized water were added respectively, and the mixture was soaked and extracted at 26°C for 28h, stirred 3 times during the period, centrifuged at 10000rpm for 10min, and filtered to obtain a fermentation liquid; wherein, the Bacillus subtilis 10 11 CFU / g, Bacillus amyloliquefaciens 10 12 CFU / g and Lactobacillus plantarum 10 11 The mass ratio of CFU / g was 1:1.25:1.5;

[0064] S4. Purification and sterilization:

[0065] The baicalein, cyclodextrin-coated chlorogenic acid and fermentation broth obtained in the above steps S1, S2 and S3 are mixed and then ultrafiltered, sterilized at 130° C. for 20 seconds, and packaged to obtain Yuhuang oral liquid.

[0066] Example 2

[0067] A method for preparing Yuhuang oral liquid for treating chicken damp-heat diarrhea comprises the following steps:

[0068] S1. Extraction of baicalin:

[0069] (1) 7.5 g of Scutellaria baicalensis was crushed into 50 mesh Scutellaria baicalensis powder, soaked in 120 g of deionized water, heated to 40° C., hydrolyzed for 14 h, and filtered to obtain a residue;

[0070] (2) adding the filter residue obtained in step (1) into ethanol with an amount 28 times the mass of the filter residue, soaking for 5 hours, filtering, and distilling the obtained filtrate under reduced pressure to obtain Scutellaria baicalensis extract;

[0071] (3) purifying the scutellaria baicalensis extract obtained in step (2) by using a macroporous resin column, and spray drying at 100° C. for 20 min to obtain baicalein; wherein the macroporous resin column is a DM101 macroporous resin column, the eluent is 90 wt % ethanol, and the volume of the macroporous resin is twice the volume of the scutellaria baicalensis extract;

[0072] S2. Encapsulated chlorogenic acid:

[0073] ① Grind 2.5 g of honeysuckle into 20-mesh honeysuckle powder, add it to the reactor, then add deionized water 10 times the mass of the honeysuckle powder, soak for 60 minutes, reflux extraction twice, the reflux extraction temperature is 100°C, and the reflux extraction time is 30 minutes each time. After the extracts are combined, they are filtered and concentrated to obtain chlorogenic acid extract;

[0074] ② Dissolve 18.5 g of β-cyclodextrin in 1000 ml of deionized water to form a β-cyclodextrin solution for later use; add the chlorogenic acid extract obtained in step ① to the reactor, and then add a β-cyclodextrin solution with a mass of 10 times that of the chlorogenic acid extract, stirring while adding, and continuing to stir for 8 hours after the addition is completed to obtain an inclusion complex;

[0075] ③ Add poloxamer 188 (4% of the mass of β-cyclodextrin used) and polyvinyl pyrrolidone (2% of the mass of β-cyclodextrin used) to the inclusion compound obtained in step ②, heat to 40°C, stir for 24h, filter, and dry at 100°C for 6h to obtain cyclodextrin-coated chlorogenic acid;

[0076] S3. Traditional Chinese medicine fermentation:

[0077] 10g of Curcuma, 5g of Codonopsis pilosula, 5g of Gardenia jasminoides, and 5g of Rhubarb were mixed and crushed into a 100-mesh powder, sterilized at 130°C for 10s, and loaded into a fermentation tank. 15g of glucose, 0.75g of urea, 0.2g of citric acid, and 20g of composite bacteria were added. Deionized water was added to adjust the water content to 40%, and the mixture was sealed and fermented at 37°C for 6 days. Then, the same amount of composite bacteria and deionized water were added respectively, and the mixture was soaked and extracted at 24°C for 30h, stirred twice, centrifuged at 8000 for 15min, and filtered to obtain a fermentation liquid. 11 CFU / g, Bacillus amyloliquefaciens 10 12 CFU / g and Lactobacillus plantarum 10 11 The mass ratio of CFU / g was 1:1:1;

[0078] S4. Purification and sterilization:

[0079] The baicalein, cyclodextrin-coated chlorogenic acid and fermentation broth obtained in the above steps S1, S2 and S3 are mixed and then ultrafiltered, sterilized at 120° C. for 30 seconds, and packaged to obtain Yuhuang oral liquid.

[0080] Example 3

[0081] A method for preparing Yuhuang oral liquid for treating chicken damp-heat diarrhea comprises the following steps:

[0082] S1. Extraction of baicalin:

[0083] (1) 13.3 g of Scutellaria baicalensis was crushed into 80 mesh Scutellaria baicalensis powder, soaked in 266 g of deionized water, heated to 60° C., hydrolyzed for 10 h, and filtered to obtain a filter residue;

[0084] (2) adding the filter residue obtained in step (1) into ethanol with an amount 32 times the mass of the filter residue, soaking for 8 hours, filtering, and distilling the obtained filtrate under reduced pressure to obtain Scutellaria baicalensis extract;

[0085] (3) Purifying the scutellaria baicalensis extract obtained in step (2) using a macroporous resin column, spray drying at 120° C. for 20 min, to obtain baicalin; wherein the macroporous resin column is a DM101 macroporous resin column, the eluent is 60 wt % ethanol, and the volume of the macroporous resin is 5 times the volume of the scutellaria baicalensis extract;

[0086] S2. Encapsulated chlorogenic acid:

[0087] ① Crush 3.4 g of honeysuckle into 50 mesh honeysuckle powder, add it to the reactor, then add deionized water 40 times the mass of the honeysuckle powder, soak for 30 minutes, reflux extraction 4 times, the reflux extraction temperature is 60 ° C, and the reflux extraction time is 90 minutes each time. The extracts are combined, filtered, and concentrated to obtain chlorogenic acid extract;

[0088] ② Dissolve 18.5g of β-cyclodextrin in 1000ml of deionized water to form a β-cyclodextrin solution for later use; add the chlorogenic acid extract obtained in step ① to the reactor, and then add a β-cyclodextrin solution with a mass of 40 times that of the chlorogenic acid extract, stirring while adding, and continue stirring for 12h after the addition is completed to obtain an inclusion complex;

[0089] ③ Add poloxamer 188 (8% of the mass of β-cyclodextrin used) and polyvinyl pyrrolidone (4% of the mass of β-cyclodextrin used) to the inclusion compound obtained in step ②, heat to 60°C, stir for 12h, filter, and dry at 80°C for 8h to obtain cyclodextrin-coated chlorogenic acid;

[0090] S3. Traditional Chinese medicine fermentation:

[0091] 20g of Curcuma, 15g of Codonopsis pilosula, 10g of Gardenia jasminoides and 10g of Rhubarb were mixed and crushed into a 50-mesh powder, sterilized at 140°C for 5s, put into a fermentation tank, added with 33g of glucose, 1.65g of urea, 0.45g of citric acid and 44g of composite bacteria, and deionized water was added to adjust the water content to 60%. The mixture was sealed and fermented at 30°C for 8 days, and then the same amount of composite bacteria and deionized water were added respectively. The mixture was soaked and extracted at 28°C for 26h, stirred 3 times during the period, centrifuged at 10000rpm for 10min, and filtered to obtain a fermentation liquid. The Bacillus subtilis 10 11 CFU / g, Bacillus amyloliquefaciens 10 12 CFU / g and Lactobacillus plantarum 10 11 The mass ratio of CFU / g was 1:1.5:2;

[0092] S4. Purification and sterilization:

[0093] The baicalein, cyclodextrin-coated chlorogenic acid and fermentation broth obtained in the above steps S1, S2 and S3 are mixed and then ultrafiltered, sterilized at 145° C. for 10 seconds, and packaged to obtain Yuhuang oral liquid.

[0094] Example 4

[0095] A method for preparing Yuhuang oral liquid for treating damp-heat diarrhea in chickens, wherein the specific implementation method is the same as that of Example 1, except that the macroporous resin column in step (3) is an AB-8 type macroporous resin column.

[0096] Example 5

[0097] A method for preparing Yuhuang oral liquid for treating damp-heat diarrhea in chickens, wherein the specific implementation method is the same as that of Example 1, except that the volume of the macroporous resin in step (3) is 1.5 times the volume of the scutellaria baicalensis extract.

[0098] Example 6

[0099] A method for preparing Yuhuang oral liquid for treating damp-heat diarrhea in chickens, wherein the specific implementation manner is the same as that of Example 1, except that the temperature of reflux extraction in step ① is 40° C., and the time of each reflux extraction is 60 minutes.

[0100] Example 7

[0101] A method for preparing Yuhuang oral liquid for treating damp-heat diarrhea in chickens, wherein the specific implementation manner is the same as that of Example 1, except that a β-cyclodextrin solution having a mass of 7.5 times that of the chlorogenic acid extract is added in step ②.

[0102] Comparative Example 1

[0103] A method for preparing Yuhuang oral liquid for treating chicken damp-heat diarrhea, the specific implementation method is the same as that of Example 1, except that 32g of Bacillus subtilis 10 11 CFU / g replaces compound bacteria.

[0104] Comparative Example 2

[0105] A method for preparing Yuhuang oral liquid for treating chicken damp-heat diarrhea comprises the following steps:

[0106] S1. 15g of Curcuma, 10g of Codonopsis pilosula, 7.5g of Gardenia jasminoides, 7.5g of Rhubarb, 3g of Honeysuckle and 10.5g of Scutellaria baicalensis were mixed and crushed into 80 mesh powder, sterilized at 135°C for 8s, loaded into a fermentation tank, 24g of glucose, 1.2g of urea, 0.3g of citric acid, 32g of composite bacteria, deionized water was added to adjust the water content to 50%, sealed and fermented at 35°C for 7 days, and then the same amount of composite bacteria and deionized water were added, and the mixture was soaked and extracted at 26°C for 28h, stirred 3 times during the period, centrifuged at 8000rpm for 15min, and filtered to obtain a fermentation broth; the Bacillus subtilis 10 11 CFU / g, Bacillus amyloliquefaciens 10 12 CFU / g and Lactobacillus plantarum 10 11 The mass ratio of CFU / g was 1:1.25:1.5;

[0107] S2. The fermentation liquid obtained in step S1 is ultrafiltered, sterilized at 135° C. for 20 seconds, and packaged to obtain Yuhuang oral liquid.

[0108] Effect evaluation:

[0109] The Yuhuang oral liquid prepared in the above Examples 1-7 and Comparative Examples 1-2 was tested, and the specific results are shown in Table 1.

[0110] Performance Test:

[0111] The experimental animals were a group of broiler chickens aged 28-30 days that had naturally fallen ill in a chicken farm and had not received any treatment. The clinical manifestations of the disease were mainly characterized by mental fatigue, standing with the head down, and yellow-white loose stools.

[0112] The above-mentioned diseased broiler chickens were randomly divided into 10 groups, with the same number of chickens in each group (250 chickens). The drinking water of the broiler chickens was prepared by mixing 2 ml of Yuhuang oral liquid with 1 L of water. The drinking water was used for 7 consecutive days. Each group had free drinking water and other conditions were the same. The 10th group was the blank control and no drugs were used. After 10 days of medication, the therapeutic effects of each group were counted. No recurrence was considered as cure. The results are shown in Table 1.

[0113] Table 1

[0114] Group Serial number Cure rate (%) mortality rate(%) Relapse and ineffectiveness / % 1 Example 1 92.8 1.6 5.6 2 Example 2 90.0 2.0 8.0 3 Example 3 90.8 2.0 7.2 4 Example 4 82.4 7.6 10.0 5 Example 5 80.4 8.4 11.2 6 Example 6 85.6 6.8 7.6 7 Example 7 83.6 7.2 9.2 8 Comparative Example 1 76.8 10.8 12.4 9 Comparative Example 2 70.8 13.6 15.6 10 Blank control 18.8 30.4 50.8

[0115] From the results in Table 1, it can be seen that the Yuhuang oral liquid prepared in Examples 1-3 has significant therapeutic effects and has good application value.

[0116] Compared with Example 1, Example 4 uses AB-8 macroporous resin different from the present invention when extracting baicalein. Compared with Example 1, Example 5 uses DM101 macroporous resin with a volume that is 1.5 times the volume of Scutellaria baicalensis extract. Both of them will reduce the extraction rate of baicalein, affect the effectiveness of Yuhuang oral liquid, and further affect the cure rate.

[0117] Compared with Example 1, when extracting chlorogenic acid, Example 6 changed the parameters of reflux extraction, resulting in a low extraction rate of chlorogenic acid; Compared with Example 1, when coating chlorogenic acid, Example 7 changed the amount of β-cyclodextrin solution added, so that the coating of β-cyclodextrin on chlorogenic acid was weakened, thereby resulting in decreased heat resistance and stability, which further affected the therapeutic rate of Yuhuang oral liquid on damp-heat diarrhea in chickens.

[0118] Compared with Example 1, Comparative Example 1 did not use composite bacteria to ferment the four traditional Chinese medicines of Curcuma, Fructus Caryophylli, Fructus Gardeniae and Rhei, thereby significantly reducing the extraction rate of the traditional Chinese medicine components.

[0119] In Comparative Example 2, all the ingredients were directly fermented, which resulted in decreased activities of baicalin and chlorogenic acid, thereby affecting the cure rate.

[0120] The above is only a preferred embodiment of the present invention and does not constitute any form of limitation to the present application. Although the present application is disclosed as above in the preferred embodiment, it is not intended to limit the present application. Any technician familiar with the profession, without departing from the scope of the technical solution of the present application, using the technical content disclosed above to make slight changes or modifications are equivalent to equivalent implementation cases. However, any simple modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution.

Claims

1. A method for preparing Yuhuang oral liquid for treating chicken damp-heat diarrhea, characterized in that: The following steps are included: S1. Extracting baicalein: purifying baicalein from Scutellaria baicalensis using a macroporous resin column; S2. Encapsulating chlorogenic acid: extracting chlorogenic acid from honeysuckle and encapsulating it with cyclodextrin to obtain cyclodextrin-encapsulated chlorogenic acid; S3. Chinese medicine fermentation: Mix turmeric, kezi, gardenia, and rhubarb, add nutrients, citric acid, and compound bacteria for fermentation to obtain a fermentation liquid; S4. Purification and sterilization: The baicalein, cyclodextrin-coated chlorogenic acid and fermentation broth obtained in the above steps S1, S2 and S3 are mixed, purified, sterilized and packaged to obtain Yuhuang oral liquid.

2. The method for preparing the Yuhuang oral liquid for treating chicken damp-heat diarrhea according to claim 1, characterized in that: The specific steps of step S1 are as follows: (1) crushing Scutellaria baicalensis into 50-80 mesh Scutellaria baicalensis powder, soaking in deionized water 16-20 times the mass of the Scutellaria baicalensis powder, heating to 40-60° C., hydrolyzing for 10-14 hours, filtering to obtain a filter residue; (2) adding the filter residue obtained in step (1) to ethanol with an amount of 28-32 times the mass of the filter residue, soaking for 5-8 hours, filtering, and distilling under reduced pressure to obtain the filtrate to obtain Scutellaria baicalensis extract; (3) Purifying the scutellaria baicalensis extract obtained in step (2) using a macroporous resin column, and drying to obtain baicalin.

3. The method for preparing the Yuhuang oral liquid for treating chicken damp-heat diarrhea according to claim 2, characterized in that: The macroporous resin column in step (3) is a DM101 macroporous resin column, and the eluent is 60-90wt% ethanol.

4. The method for preparing the Yuhuang oral liquid for treating chicken damp-heat diarrhea according to claim 2, characterized in that: In step (3), the volume ratio of the scutellaria baicalensis extract to the macroporous resin in the macroporous resin column is 1:(2-5).

5. The method for preparing the Yuhuang oral liquid for treating chicken damp-heat diarrhea according to claim 1, characterized in that: The specific steps of step S2 are as follows: ① Crush the honeysuckle into 20-50 mesh honeysuckle powder, add it to the reactor, then add deionized water 10-40 times the mass of the honeysuckle powder, soak for 30-60 minutes, reflux extraction 2-4 times, combine the extracts, filter, and concentrate to obtain chlorogenic acid extract; ② Dissolve cyclodextrin in deionized water to form a 16-20 g / L cyclodextrin solution, add the chlorogenic acid extract obtained in step ①, and stir while adding to obtain an inclusion complex; ③ Add the surfactant and stabilizer to the inclusion compound obtained in step ②, heat to 40-60° C., stir for 12-24 hours, filter, and dry to obtain cyclodextrin-coated chlorogenic acid.

6. The method for preparing the Yuhuang oral liquid for treating chicken damp-heat diarrhea according to claim 5, characterized in that: The temperature of the reflux extraction in step ① is 60-100° C., and the time of each reflux extraction is 30-90 min.

7. The method for preparing the Yuhuang oral liquid for treating chicken damp-heat diarrhea according to claim 5, characterized in that: The mass ratio of the chlorogenic acid extract to the cyclodextrin solution in step ② is 1:(10-40).

8. The method for preparing the Yuhuang oral liquid for treating chicken damp-heat diarrhea according to claim 1, characterized in that: The specific steps of step S3 are as follows: In parts by weight, 10-20 parts of turmeric, 5-15 parts of kohlrabi, 5-10 parts of gardenia and 5-10 parts of rhubarb are mixed and crushed, sterilized, and loaded into a fermentation tank, and a carbon source, a nitrogen source, citric acid and composite bacteria are added, and deionized water is added to adjust the water content to 40-60%, and the mixture is sealed and fermented at 30-37°C for 6-8 days, and then the same amount of composite bacteria and deionized water are added, and the mixture is soaked and extracted at 24-28°C for 26-30 hours, during which the mixture is stirred 2-3 times, centrifuged, filtered, and fermented to obtain a fermentation liquid.

9. The method for preparing the Yuhuang oral liquid for treating chicken damp-heat diarrhea according to claim 1, characterized in that: The composite bacteria in step S3 is Bacillus subtilis 10 9-12 CFU / g, Bacillus amyloliquefaciens 10 9-12 CFU / g and lactic acid bacteria 10 10-11 CFU / g.

10. The method for preparing Yuhuang oral liquid for treating chicken damp-heat diarrhea according to claim 1, characterized in that: The mass ratio of honeysuckle, scutellaria baicalensis and curcuma is 1:(3-4):(4-6).