Method for extracting ovateleaf holly bark medicinal material and ovateleaf holly bark extraction powder

Abstract

The invention discloses an extraction method of an ovateleaf holly bark medicinal material and ovateleaf holly bark extraction powder. On one hand, the extraction method of the ovateleaf holly bark medicinal material provided by the invention comprises the following steps: S1, taking the ovateleaf holly bark medicinal material, and performing crushing treatment; s2, performing extraction treatment on the medicinal material powder obtained in the step S1 by taking ethanol as an extraction solvent and enzyme as an extraction auxiliary agent and adopting a biological enzyme method; s3, performing filtration and ultrafiltration membrane concentration treatment on the extraction mixed solution obtained in the step S2 to obtain an ovateleaf holly bark concentrated solution; and S4, performing spray drying treatment on the ovateleaf holly bark concentrated solution obtained in S3 to obtain the ovateleaf holly bark extract. On the other hand, the ovateleaf holly bark extraction powder comprises the ovateleaf holly bark extract prepared by the extraction method of the ovateleaf holly bark medicinal material, and auxiliary materials. The process steps are relatively simple, the extraction rate of the effective components of the ovateleaf holly bark is relatively high, the energy consumption is low, the speed is high, and the quality of the extracted ovateleaf holly bark extraction powder is very stable.

Description

Technical Field

[0001] The present application relates to the field of pharmaceutical technologies, and in particular, to an extraction method for Ilex rotunda Thunb. and an Ilex rotunda Thunb. extract powder. Background Art

[0002] Ilex rotunda Thunb. is a traditional Chinese medicine, which is the bark or root bark of the plant Ilex rotunda Thunb. of the Aquifoliaceae family. Ilex rotunda Thunb. is bitter and cool in nature, and has the effects of clearing heat and detoxifying, cooling blood and stopping bleeding, promoting qi circulation and relieving pain, etc. It is mostly used for cold fever, rash fever, tonsillitis, sore throat, breast carbuncle, abdominal distension pain, diarrhea, dysentery, acute gastroenteritis, rheumatic bone pain; externally used for treating traumatic injury, external bleeding, burns and scalds, etc. Now it is mostly used in veterinary clinics for the prevention and treatment of intestinal diseases.

[0003] In the mid-1950s of the 20th century, Zhu Renhong first decomposed syringenin A and syringenin B from Ilex rotunda Thunb.; subsequently, Xie Peishan et al. confirmed that the syringenin B isolated by them and syringenin A of Zhu were both syringin, and syringenin B was consistent with the bitter substance pedunculoside isolated from Ilex rotunda Thunb. reported by Haseyama Tsunao et al. in the early 1970s in Japan. Haseyama Tsunao et al. also confirmed that its aglycone was rotundic acid (ilexol). Existing technical research has found that the extraction of Ilex rotunda Thunb. and its medicated serum have good effects on Escherichia coli producing ESBLs. Especially after combining with antibacterial drugs, the antibacterial effect on drug-resistant Escherichia coli is enhanced, and the research mechanism finds that it has a destructive effect on the cell wall.

[0004] In the production of traditional Chinese veterinary medicine, the Ilex rotunda Thunb. used is mostly unprocessed traditional Chinese medicine. Unprocessed traditional Chinese medicine often cannot exert its due medicinal effects. The use of many traditional Chinese medicines without processing will lead to an increase in toxicity, such as Evodia rutaecarpa, Genkwa flos, etc.; in addition, traditional Chinese medicines may contain relatively high pesticide residues, heavy metals or other pollutants, which will affect the quality stability and clinical efficacy of traditional Chinese medicine preparations. The processing of traditional Chinese medicine has the advantages of improving medicinal effects, high safety, and enhancing clinical efficacy. The modern extraction process of traditional Chinese medicine can maximize the extraction of the effective components in Ilex rotunda Thunb., so that the effects of Ilex rotunda Thunb. can be maximally exerted.

[0005] Traditional Chinese medicine extraction methods such as water extraction, decoction, cold maceration, percolation, etc. all have problems such as long cycle, low extraction rate of active ingredients, and low utilization rate of medicinal materials. Traditional concentration methods such as vacuum concentration and reduced-pressure concentration consume a large amount of energy, take a long time, and are not friendly to thermosensitive components in medicinal materials. For the extraction of Ilex rotunda Thunb. in the prior art, the process method recorded in the literature titled "Research on the Production Process of Ilex rotunda Thunb. Extract", jointly developed by the National Engineering Research Center for Modernization of Chinese Medicine Extraction and Separation Process and Guangzhou Baiyunshan Hanfang Modern Pharmaceutical Co., Ltd. and published in 2019, is mainly adopted. This process method uses ethanol as the extraction solvent and optimizes the relevant parameters of the extraction process. However, the extraction rate of the active ingredients of Ilex rotunda Thunb. by the above existing process is not high. Summary of the Invention

[0006] In order to solve at least one of the above technical problems and develop an extraction process with relatively simple process steps, high extraction rate of active ingredients of Ilex rotunda Thunb., low energy consumption, fast rate, and very stable quality of the Ilex rotunda Thunb. extract prepared by extraction, the present application provides an extraction method for Ilex rotunda Thunb. medicinal materials and Ilex rotunda Thunb. extract powder.

[0007] On the one hand, the present application provides an extraction method for Ilex rotunda Thunb. medicinal materials, including the following steps:

[0008] S1. Take Ilex rotunda Thunb. medicinal materials, crush them to obtain medicinal material powder;

[0009] S2. Use the medicinal material powder obtained in step S1, use ethanol as the extraction solvent, use an enzyme as an extraction adjuvant, and adopt a biological enzyme method for extraction treatment to obtain an extraction mixture;

[0010] S3. Filter and ultrafiltration membrane concentrate the extraction mixture obtained in step S2 to obtain an Ilex rotunda Thunb. concentrate;

[0011] S4. Spray-dry the Ilex rotunda Thunb. concentrate obtained in S3 to prepare an Ilex rotunda Thunb. extract.

[0012] Optionally, in step S1, the Ilex rotunda Thunb. medicinal materials are processed medicinal materials.

[0013] Optionally, in step S1, for the crushing treatment, the Ilex rotunda Thunb. medicinal materials need to be crushed into medicinal material powder with a particle size of 24-80 mesh.

[0014] Optionally, in step S2, the ethanol used is an ethanol solution with a volume fraction of 20-70%, and the material-liquid ratio of the medicinal material powder to the extraction solvent is 1:20-30.

[0015] Preferably, the pH value of the ethanol solution is controlled at 4.5-5.5.

[0016] More preferably, citric acid is used as a regulator to adjust the pH value of the ethanol solution.

[0017] Optionally, in the step S2, the addition amount of the enzyme accounts for 1-4% of the mass of the medicinal material powder.

[0018] Further optionally, in the step S2, the enzyme used is a composite enzyme, and the composite enzyme includes cellulase and pectinase; preferably, the mass ratio of the cellulase to the pectinase is 1:2.

[0019] Further optionally, in the step S2, the composite enzyme includes cellulase, pectinase and neutral protease; preferably, the mass ratio of the cellulase, the pectinase and the neutral protease is 1:2:1.

[0020] Optionally, in the step S2, the extraction temperature of the bio-enzymatic extraction treatment is controlled at 45-60°C, and the extraction time is 90-200 min.

[0021] Preferably, in the step S3, the operating pressure of the ultrafiltration membrane concentration is 0.4 MPa, and the concentration is carried out until the solid content is 20%-30%.

[0022] Optionally, in the step S4, before the spray drying treatment, an adjuvant needs to be added to the Evodia rutaecarpa concentrated liquid, and the adjuvant is selected from one or more of Tween-80, sodium alginate, xanthan gum, PEG400, PVPK30 and glycerol.

[0023] Preferably, the addition amount of the adjuvant accounts for 1-5% of the total mass of the Evodia rutaecarpa concentrated liquid.

[0024] Preferably, the inlet air temperature during the spray drying is 150-200°C; the material temperature is 60-80°C; the outlet air temperature is 70-100°C.

[0025] On the other hand, the present application also provides an Evodia rutaecarpa extraction powder, which includes the Evodia rutaecarpa extract prepared by the extraction method of the above-mentioned Evodia rutaecarpa medicinal material and excipients.

[0026] Preferably, the excipients are selected from one or more of anhydrous glucose, soluble starch, and dextrin.

[0027] In summary, the present invention includes at least one of the following beneficial technical effects:

[0028] 1. The extraction method of the present application adopts an ethanol-based extraction solvent and a bio-enzymatic extraction method assisted by bio-enzymes, which can effectively improve the transfer rate of the active ingredients in the Evodia rutaecarpa medicinal material during extraction, and can accelerate the dissolution of the active ingredients in the Evodia rutaecarpa medicinal material, can greatly improve the utilization rate of the Evodia rutaecarpa medicinal material, and effectively improve the quality of the medicinal material extract.

[0029] 2. The extraction method of the present application adopts the process steps of membrane concentration combined with spray drying, and the obtained Herba Cibotii extract has excellent quality and good stability.

[0030] 3. The present application uses the Herba Cibotii extract obtained by a specific extraction method as raw material, mixes it with auxiliary materials, and prepares a single Herba Cibotii extract powder, which has good stability and can be used as a veterinary drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 A schematic diagram of a process for preparing the Jiubing extract powder according to an embodiment of the present application;

[0032] Figure 2 This is the liquid chromatography detection spectrum of the Herba Lysimachiae extract of Example 2 of the present application;

[0033] Figure 3 This is the liquid chromatography detection spectrum of the Herba Lysimachiae extract of Example 3 of the present application;

[0034] Figure 4 This is the liquid chromatography detection spectrum of the Herba Cibotii extract of Example 4 of the present application. DETAILED DESCRIPTION

[0035] The present application is further described in detail below with reference to the accompanying drawings and embodiments.

[0036] The present application proposes a method for extracting the medicinal material of Jiubiying, comprising the following steps:

[0037] S1. Take Jiubing medicinal materials, crush them, and obtain medicinal material powder;

[0038] S2, extracting the medicinal material powder obtained in step S1 by bio-enzyme method using ethanol as extraction solvent and enzyme as extraction auxiliary agent to obtain an extraction mixture;

[0039] S3, filtering and concentrating the extracted mixed solution obtained in step S2 through an ultrafiltration membrane to obtain a concentrated solution of Jiubing;

[0040] S4, spray-drying the scutellaria baicalensis concentrated solution obtained in S3 to obtain a scutellaria baicalensis extract.

[0041] The present application also designs a scutellaria extract powder, comprising a scutellaria extract obtained by the above-mentioned scutellaria extracting method, and an auxiliary material. The auxiliary material can be selected from one or more of anhydrous glucose, soluble starch or dextrin. The scutellaria extract powder of the present application can be prepared by mixing the scutellaria extract with the auxiliary material, and the drug has good stability and can be used as a veterinary drug.

[0042] The following are examples of the present application:

[0043] For the Herba Ilicis Rotundae used in the embodiments of the present application, refer to the processing method of Herba Ilicis Rotundae recorded in the Chinese Pharmacopoeia (2020 Edition): remove impurities, wash, moisten thoroughly, slice, and then perform processing.

[0044] Example 1

[0045] As Figure 1 shown, the extraction method of Herba Ilicis Rotundae in this embodiment includes the following steps:

[0046] S1. Take the Herba Ilicis Rotundae, perform comminution treatment, comminute to a particle size of 24 mesh to obtain the medicinal material powder;

[0047] S2. Take a 20% ethanol solution, add citric acid to adjust the pH value to 4.5 as the extraction solvent; take cellulase and pectinase, mix them according to a mass ratio of 1:2 as the enzyme; according to the material-liquid ratio of the medicinal material powder to the extraction solvent of 1:20 and the enzyme addition amount accounting for 1.0% of the mass of the medicinal material powder, add the medicinal material powder, extraction solvent and enzyme into the extraction unit, control the extraction temperature at 45°C and the time at 90 min, and perform extraction treatment to obtain the extraction mixture;

[0048] S3. Filter the extraction mixture obtained in step S2, and then concentrate the filtrate by ultrafiltration membrane to obtain the Herba Ilicis Rotundae concentrated solution; the operating pressure of ultrafiltration membrane concentration is 0.4 MPa, and concentrate to a solid content of 20%;

[0049] S4. Add 1% of PVPK30 to the Herba Ilicis Rotundae concentrated solution obtained in S3, and perform spray drying treatment to obtain the Herba Ilicis Rotundae extract; the inlet air temperature of the spray drying treatment is 150°C, the material temperature is 60°C, and the outlet air temperature is 70°C.

[0050] The Herba Ilicis Rotundae extraction powder in this embodiment uses the Herba Ilicis Rotundae extract prepared by the above extraction method as the main raw material and anhydrous glucose as the auxiliary material; the raw material and the auxiliary material are fully mixed according to a mass ratio of 1:2 and then made.

[0051] Example 2

[0052] The extraction method of Herba Ilicis Rotundae in this embodiment includes the following steps:

[0053] S1. Take the Herba Ilicis Rotundae, perform comminution treatment, comminute to a particle size of 50 mesh to obtain the medicinal material powder;

[0054] S2. Take a 30% ethanol solution, add citric acid to adjust the pH value to 5.0 as the extraction solvent; take cellulase and pectinase, mix them according to a mass ratio of 1:2 as the enzyme; according to the material-liquid ratio of medicinal powder to extraction solvent of 1:20 and the enzyme addition amount accounting for 1.5% of the mass of the medicinal powder, add the medicinal powder, extraction solvent and enzyme into the extraction unit, control the extraction temperature at 48 °C and the time at 120 min for extraction treatment to obtain an extraction mixture;

[0055] S3. Filter the extraction mixture obtained in step S2, and then concentrate the filtrate by ultrafiltration membrane to obtain the Evodia extract concentrate; the operating pressure of ultrafiltration membrane concentration is 0.4 MPa, and concentrate to a solid content of 25%;

[0056] S4. Add 2% Tween-80 to the Evodia extract concentrate obtained in S3, and carry out spray drying treatment to obtain the Evodia extract; the inlet air temperature of the spray drying treatment is 160 °C, the material temperature is 65 °C, and the outlet air temperature is 75 °C.

[0057] The Evodia extraction powder of this example uses the Evodia extract prepared by the above extraction method as the main raw material and soluble starch as the auxiliary material; the raw material and the auxiliary material are fully mixed according to a mass ratio of 1:2 and then made.

[0058] Example 3

[0059] The extraction method of the Evodia medicinal materials in this example includes the following steps:

[0060] S1. Take the Evodia medicinal materials, crush them to a particle size of 65 mesh to obtain medicinal powder;

[0061] S2. Take a 40% ethanol solution, add citric acid to adjust the pH value to 5.0 as the extraction solvent; take cellulase and pectinase, mix them according to a mass ratio of 1:2 as the enzyme; according to the material-liquid ratio of medicinal powder to extraction solvent of 1:25 and the enzyme addition amount accounting for 2.0% of the mass of the medicinal powder, add the medicinal powder, extraction solvent and enzyme into the extraction unit, control the extraction temperature at 50 °C and the time at 150 min for extraction treatment to obtain an extraction mixture;

[0062] S3. Filter the extraction mixture obtained in step S2, and then concentrate the filtrate by ultrafiltration membrane to obtain the Evodia extract concentrate; the operating pressure of ultrafiltration membrane concentration is 0.4 MPa, and concentrate to a solid content of 20%;

[0063] S4. Add 1% Tween-80 and 2% PEG400 to the Evodia extract concentrate obtained in S3, and carry out spray drying treatment to obtain the Evodia extract; the inlet air temperature of the spray drying treatment is 170 °C, the material temperature is 70 °C, and the outlet air temperature is 80 °C.

[0064] The Jiebi ying extract powder of this example is made with the Jiebi ying extract prepared by the above extraction method as the main raw material and dextrin as the auxiliary material; the raw material and the auxiliary material are fully mixed according to a mass ratio of 1:2.

[0065] Example 4

[0066] The extraction method of the Jiebi ying medicinal material in this example includes the following steps:

[0067] S1. Take the Jiebi ying medicinal material, crush it, and crush it to a particle size of 80 mesh to obtain the medicinal material powder.

[0068] S2. Take a 40% ethanol solution, add citric acid to adjust the pH value to 5.5 as the extraction solvent; take cellulase and pectinase, mix them according to a mass ratio of 1:2 as the enzyme; according to the material-liquid ratio of the medicinal material powder to the extraction solvent of 1:30 and the enzyme addition amount accounting for 2.5% of the mass of the medicinal material powder, add the medicinal material powder, the extraction solvent and the enzyme into the extraction unit, control the extraction temperature at 55 °C and the time at 180 min for extraction treatment to obtain the extraction mixture.

[0069] S3. Filter the extraction mixture obtained in step S2, and then concentrate the filtrate by ultrafiltration membrane to obtain the Jiebi ying concentrated solution; the operating pressure of the ultrafiltration membrane concentration is 0.4 MPa, and it is concentrated to a solid content of 20%.

[0070] S4. Add 2% Tween-80 and 2% glycerol to the Jiebi ying concentrated solution obtained in S3, and carry out spray drying treatment to prepare the Jiebi ying extract; the inlet air temperature of the spray drying treatment is 180 °C, the material temperature is 75 °C, and the outlet air temperature is 80 °C.

[0071] The Jiebi ying extract powder of this example is made with the Jiebi ying extract prepared by the above extraction method as the main raw material and anhydrous glucose as the auxiliary material; the raw material and the auxiliary material are fully mixed according to a mass ratio of 1:2.

[0072] Example 5

[0073] The extraction method of the Jiebi ying medicinal material in this example includes the following steps:

[0074] S1. Take the Jiebi ying medicinal material, crush it, and crush it to a particle size of 65 mesh to obtain the medicinal material powder.

[0075] S2. Take a 70% ethanol solution, add citric acid to adjust the pH value to 4.8 as the extraction solvent; take cellulase and pectinase, mix them according to a mass ratio of 1:2 as the enzyme; according to the material-liquid ratio of the medicinal material powder to the extraction solvent of 1:25 and the enzyme addition amount accounting for 4.0% of the mass of the medicinal material powder, add the medicinal material powder, the extraction solvent and the enzyme into the extraction unit, control the extraction temperature at 60 °C and the time at 200 min for extraction treatment to obtain an extraction mixture;

[0076] S3. Filter the extraction mixture obtained in step S2, and then concentrate the filtrate by ultrafiltration membrane to obtain the Evodia extract; the operating pressure for ultrafiltration membrane concentration is 0.4 MPa, and concentrate to a solid content of 25%;

[0077] S4. Add 2.5% Tween-80 and 2.5% xanthan gum to the Evodia extract obtained in S3, and carry out spray drying treatment to prepare the Evodia extract; the inlet air temperature for spray drying treatment is 200 °C, the material temperature is 80 °C, and the outlet air temperature is 100 °C.

[0078] The Evodia extraction powder of this example uses the Evodia extract prepared by the above extraction method as the main raw material and anhydrous glucose as the auxiliary material; the raw material and the auxiliary material are fully mixed according to a mass ratio of 1:2 and then made.

[0079] The following are the comparative examples of this application. Comparative examples 1-3 of this application all adopt the existing extraction process.

[0080] Comparative example 1

[0081] The extraction method of the Evodia medicinal material in this comparative example includes the following steps:

[0082] Take the prescribed amount of Evodia medicinal material, add 10 times the amount of water, decoct 3 times in total, each time for 120 minutes, filter the extraction liquid, combine the extraction liquids, concentrate under reduced pressure to a solid content of 30%, and spray dry to obtain the Evodia extract.

[0083] According to the method of Example 4, mix the Evodia extract with anhydrous glucose to prepare the Evodia extraction powder.

[0084] Comparative example 2

[0085] The extraction method of the Evodia medicinal material in this comparative example includes the following steps:

[0086] Take the prescribed amount of Evodia medicinal material, pulverize it to 200 mesh, add 10 times the amount of 20% ethanol, extract 3 times in total, each time for 30 minutes, filter the extraction liquid, combine the extractions, concentrate under reduced pressure to a solid content of 20%, and spray dry to obtain the Evodia extract.

[0087] According to the method of Example 4, the extract of Ilex rotunda Thunb. was mixed with anhydrous glucose to prepare the extract powder of Ilex rotunda Thunb.

[0088] Comparative Example 3

[0089] This application takes the optimal extraction process method recorded in the journal literature titled "Research on the Production Process of the Extract of Ilex rotunda Thunb.", published in the 4th issue of the 47th volume of "Guangzhou Chemical Industry" in February 2019, as the production process of the extract of Ilex rotunda Thunb. in Comparative Example 3 of this application.

[0090] According to the method of Example 4, the extract of Ilex rotunda Thunb. was mixed with anhydrous glucose to prepare the extract powder of Ilex rotunda Thunb.

[0091] Detection

[0092] 1. With the two main components of ilicinin and syringin as references, the transfer rate of the active ingredients was detected and calculated. The transfer rate calculation formula: transfer rate = the amount of the target component in the extract of Ilex rotunda Thunb. / the amount of the target component in the medicinal material of Ilex rotunda Thunb.×100%.

[0093] The amount of the target component in the extract of Ilex rotunda Thunb. was determined by the HPLC method recorded in the Chinese Pharmacopoeia (2020 Edition), and the amount of the target component in the medicinal material of Ilex rotunda Thunb. was determined by the detection method recorded in the Chinese Pharmacopoeia (2020 Edition).

[0094] 2. Dissolution rate of the extract powder of Ilex rotunda Thunb.

[0095] Determination method: Weigh 2 g of the extract powder of Ilex rotunda Thunb., add 50 ml of water, stir and dissolve, and record its dissolution rate.

[0096] 3. Stability of the extract powder of Ilex rotunda Thunb.

[0097] The extract powder samples of Ilex rotunda Thunb. were packaged with polyester / aluminum / polyethylene composite film / bag for pharmaceutical packaging, and stability tests were carried out.

[0098] Investigation conditions: In accordance with the "Guidelines for the Stability Testing of Raw Materials and Preparations" in Part I of the Chinese Veterinary Pharmacopoeia (2020 Edition), the contents of ilicinin and syringin were determined by the HPLC method, and the specific detection steps were carried out with reference to the content of Announcement No. 751 of the Ministry of Agriculture and Rural Affairs.

[0099] The detection results are shown in Tables 1 to 3 below, and the HPLC detection spectra of the extracts of Ilex rotunda Thunb. prepared in Examples 2 to 4 are as Figures 2 - 4 shown.

[0100] Table 1 Detection results of the transfer rate of Examples 1 to 5 and Comparative Examples 1 to 3

[0101]

[0102] Table 2 Dissolution rate of the extract powder of Ilex rotunda Thunb.

[0103] Dissolution rate (s) Example 1 18 Example 2 16 Example 3 18 Example 4 19 Example 5 17 Comparative Example 1 84 Comparative Example 2 90 Comparative Example 3 80

[0104] Table 3 Stability test results of Examples 1-5 and Comparative Examples 1-3

[0105]

[0106]

[0107] From the data in Tables 1-3 and Figures 2 - 4 the detection spectra, it can be seen that for the extraction processes of Examples 1-5 of the present application, compared with the extraction processes of Comparative Examples 1-3, the transfer rate of the active ingredient has been significantly increased, the dissolution rate of the Folium Illicii Extract powder prepared from the Folium Illicii Extract obtained by extraction has been significantly accelerated, and the stability has also been significantly improved.

[0108] Clinical efficacy test:

[0109] Take the Folium Illicii Extract powder samples prepared by the methods of Examples 1-5 of the present application and Comparative Examples 1-3, and conduct a clinical efficacy test for treating livestock with Escherichia coli disease.

[0110] 1. Test grouping and administration method

[0111] Take 100 healthy chickens as the negative control, which is experimental group 0; take 100 diseased chickens as the positive control, which is experimental group 9. Another 800 diseased chickens are randomly divided into 8 groups, namely experimental groups 1-8, with 100 chickens in each group; the test chickens are allowed to feed freely, and the administration method is to drink throughout the day. The drugs are added to 1 L of water according to the grouping, and the drugs are administered continuously for 7 days. After the treatment, the cure situation of the chicken flock is detected. The drugs used in experimental groups 1-5 are the Folium Illicii Extract powder prepared by the methods of Examples 1-5, and the drugs used in experimental groups 6-8 are the Folium Illicii Extract powder prepared by the methods of Comparative Examples 1-3.

[0112] 2. Efficacy determination and result analysis

[0113] Cured: Clinical signs are normal; Cure rate = number of cured chickens in the group / total number of chickens in the group × 100%

[0114] Effective: The clinical symptoms of the surviving chickens are alleviated;

[0115] Effective rate = number of effective chickens in the group / total number of chickens in the group × 100%

[0116] Total effective rate = (number of cured chickens in the group + number of effective chickens in the group) / total number of chickens in the group × 100%

[0117] Ineffective: The clinical symptoms of the surviving chickens are not alleviated and even die. Ineffective rate = number of ineffective chickens in the group / total number of chickens in the group × 100%

[0118] At the end of the experiment on the 8th day after drug withdrawal, the chicken flock was detected to determine the curative effect. The results are shown in Table 4 below.

[0119] Table 4 Clinical trial results

[0120] Grouping Number of cured (pcs) Number of effective (pcs) Number of ineffective (pcs) Total effective rate (%) Group 0 (negative control) 100 0 0 100.00 Group 1 (Example 1) 42 39 17 81.00 Group 2 (Example 2) 45 40 15 85.00 Group 3 (Example 3) 47 42 11 89.00 Group 4 (Example 4) 41 40 19 81.00 Group 5 (Example 5) 48 45 7 93.00 Group 6 (Comparative Example 1) 19 24 57 43.00 Group 7 (Comparative Example 2) 23 26 51 49.00 Group 8 (Comparative Example 3) 25 30 45 55.00 Group 9 (positive control) 0 0 100 0.00

[0121] According to the data in Table 4, the Folium Ilicis Rotundatae extraction powder prepared according to Examples 1-5 of the present application, and the Folium Ilicis Rotundatae extraction powder prepared according to Comparative Examples 1-3, all have obvious therapeutic effects on chicken Escherichia coli disease. However, the Folium Ilicis Rotundatae extraction powder prepared according to Examples 1-5 of the present application is significantly more prominent in terms of treatment effective rate and cure rate, and its total effective rate exceeds 80%; while the Folium Ilicis Rotundatae extraction powder prepared according to Comparative Examples 1-3 has a total effective rate of no more than 55%. Thus, it can be seen that the Folium Ilicis Rotundatae extraction powder prepared in the present application is significantly superior to the products of the comparative examples in the treatment effect on chicken Escherichia coli disease.

[0122] In addition, according to the data in Table 4, it can also be seen that the treatment effect of Example 5 of the present application is the best, while the treatment effects of Examples 1 and 4 are relatively weaker. It can be seen that the preparation process of the present application has a more excellent treatment effect when the transfer rates of ilicinin and syringin are higher.

[0123] On this basis, the applicant further studied and optimized the addition of enzymes; and further studied and optimized the addition of auxiliaries for spray drying.

[0124] The following are Examples 6-14 of the present application, and the extraction process parameters refer to Example 3.

[0125] Example 6

[0126] The extraction method of the Folium Ilicis Rotundatae medicinal material in this example includes the following steps:

[0127] S1. Take the Folium Ilicis Rotundatae medicinal material, crush it, and crush it to a particle size of 65 mesh to obtain a medicinal material powder.

[0128] S2. Take a 40% ethanol solution, add citric acid to adjust the pH value to 5.0 as the extraction solvent; take cellulase, pectinase and neutral protease, mix them according to a mass ratio of 1:1:1 as the enzyme; according to the material-liquid ratio of the medicinal material powder and the extraction solvent of 1:25 and the enzyme addition amount accounting for 2.0% of the mass of the medicinal material powder, add the medicinal material powder, the extraction solvent and the enzyme into the extraction unit, control the extraction temperature at 50°C, and the time at 150 min, and perform extraction treatment to obtain an extraction mixture.

[0129] S3. Filter the extraction mixture obtained in step S2, and then concentrate the filtrate by ultrafiltration membrane to obtain the Evodia lepta concentrate. The operating pressure for ultrafiltration membrane concentration is 0.4 MPa, and it is concentrated to a solid content of 20%.

[0130] S4. Add 1% Tween-80 and 2% PEG400 to the Evodia lepta concentrate obtained in S3, and then perform spray drying to obtain the Evodia lepta extract. The inlet air temperature for spray drying is 170 °C, the material temperature is 70 °C, and the outlet air temperature is 80 °C.

[0131] Example 7

[0132] The extraction method of Evodia lepta medicinal materials in this example includes the following steps:

[0133] S1. Take Evodia lepta medicinal materials, crush them to a particle size of 65 mesh to obtain medicinal material powder.

[0134] S2. Take 40% ethanol solution, add citric acid to adjust the pH value to 5.0 as the extraction solvent. Take cellulase, pectinase and neutral protease, mix them according to the mass ratio of 2:1:1 as the enzyme. According to the material-liquid ratio of medicinal material powder to extraction solvent of 1:25 and the enzyme addition amount accounting for 2.0% of the mass of the medicinal material powder, add the medicinal material powder, extraction solvent and enzyme into the extraction unit, control the extraction temperature at 50 °C and the time at 150 min for extraction to obtain the extraction mixture.

[0135] S3. Filter the extraction mixture obtained in step S2, and then concentrate the filtrate by ultrafiltration membrane to obtain the Evodia lepta concentrate. The operating pressure for ultrafiltration membrane concentration is 0.4 MPa, and it is concentrated to a solid content of 20%.

[0136] S4. Add 1% Tween-80 and 2% PEG400 to the Evodia lepta concentrate obtained in S3, and then perform spray drying to obtain the Evodia lepta extract. The inlet air temperature for spray drying is 170 °C, the material temperature is 70 °C, and the outlet air temperature is 80 °C.

[0137] Example 8

[0138] The extraction method of Evodia lepta medicinal materials in this example includes the following steps:

[0139] S1. Take Evodia lepta medicinal materials, crush them to a particle size of 65 mesh to obtain medicinal material powder.

[0140] S2. Take a 40% ethanol solution, add citric acid to adjust the pH value to 5.0 as the extraction solvent; take cellulase, pectinase and neutral protease, mix them according to the mass ratio of 1:1:2 as the enzyme; according to the material-liquid ratio of the medicinal material powder to the extraction solvent of 1:25 and the enzyme addition amount accounting for 2.0% of the mass of the medicinal material powder, add the medicinal material powder, the extraction solvent and the enzyme into the extraction unit, control the extraction temperature at 50 °C and the time at 150 min, and carry out the extraction treatment to obtain an extraction mixture;

[0141] S3. Filter the extraction mixture obtained in step S2, and then concentrate the filtrate by ultrafiltration membrane to obtain the Evodia rutaecarpa concentrate; the operating pressure of ultrafiltration membrane concentration is 0.4 MPa, and it is concentrated to a solid content of 20%;

[0142] S4. Add 1% Tween-80 and 2% PEG400 to the Evodia rutaecarpa concentrate obtained in S3, and carry out spray drying treatment to obtain the Evodia rutaecarpa extract; the inlet air temperature of the spray drying treatment is 170 °C, the material temperature is 70 °C, and the outlet air temperature is 80 °C.

[0143] Example 9

[0144] The extraction method of Evodia rutaecarpa medicinal materials in this example includes the following steps:

[0145] S1. Take Evodia rutaecarpa medicinal materials, crush them to a particle size of 65 mesh to obtain medicinal material powder;

[0146] S2. Take a 40% ethanol solution, add citric acid to adjust the pH value to 5.0 as the extraction solvent; take cellulase, pectinase and neutral protease, mix them according to the mass ratio of 1:2:1 as the enzyme; according to the material-liquid ratio of the medicinal material powder to the extraction solvent of 1:25 and the enzyme addition amount accounting for 2.0% of the mass of the medicinal material powder, add the medicinal material powder, the extraction solvent and the enzyme into the extraction unit, control the extraction temperature at 50 °C and the time at 150 min, and carry out the extraction treatment to obtain an extraction mixture;

[0147] S3. Filter the extraction mixture obtained in step S2, and then concentrate the filtrate by ultrafiltration membrane to obtain the Evodia rutaecarpa concentrate; the operating pressure of ultrafiltration membrane concentration is 0.4 MPa, and it is concentrated to a solid content of 20%;

[0148] S4. Add 1% Tween-80 and 2% PEG400 to the Evodia rutaecarpa concentrate obtained in S3, and carry out spray drying treatment to obtain the Evodia rutaecarpa extract; the inlet air temperature of the spray drying treatment is 170 °C, the material temperature is 70 °C, and the outlet air temperature is 80 °C.

[0149] Example 10

[0150] The extraction method of the medicinal material of Ilex rotunda Thunb. in this embodiment comprises the following steps:

[0151] S1. Take the medicinal material of Ilex rotunda Thunb., carry out comminution treatment, comminute it to a particle size of 65 mesh to obtain medicinal material powder;

[0152] S2. Take a 40% ethanol solution, add citric acid to adjust the pH value to 5.0 as the extraction solvent; take cellulase, pectinase and neutral protease, mix them according to a mass ratio of 2:2:1 as the enzyme; according to the material-liquid ratio of the medicinal material powder to the extraction solvent of 1:25 and the enzyme addition amount accounting for 2.0% of the mass of the medicinal material powder, add the medicinal material powder, the extraction solvent and the enzyme into the extraction unit, control the extraction temperature at 50 °C and the time at 150 min, carry out extraction treatment to obtain an extraction mixture;

[0153] S3. Filter the extraction mixture obtained in step S2, and then concentrate the filtrate by ultrafiltration membrane to obtain the concentrated liquid of Ilex rotunda Thunb.; the operating pressure of ultrafiltration membrane concentration is 0.4 MPa, and concentrate it to a solid content of 20%;

[0154] S4. Add 1% of Tween-80 and 2% of PEG400 to the concentrated liquid of Ilex rotunda Thunb. obtained in S3, and carry out spray drying treatment to prepare the extract of Ilex rotunda Thunb.; the inlet air temperature of the spray drying treatment is 170 °C, the material temperature is 70 °C, and the outlet air temperature is 80 °C.

[0155] Example 11

[0156] The extraction method of the medicinal material of Ilex rotunda Thunb. in this embodiment comprises the following steps:

[0157] S1. Take the medicinal material of Ilex rotunda Thunb., carry out comminution treatment, comminute it to a particle size of 65 mesh to obtain medicinal material powder;

[0158] S2. Take a 40% ethanol solution, add citric acid to adjust the pH value to 5.0 as the extraction solvent; take cellulase, pectinase and neutral protease, mix them according to a mass ratio of 1:2:1 as the enzyme; according to the material-liquid ratio of the medicinal material powder to the extraction solvent of 1:25 and the enzyme addition amount accounting for 2.0% of the mass of the medicinal material powder, add the medicinal material powder, the extraction solvent and the enzyme into the extraction unit, control the extraction temperature at 50 °C and the time at 150 min, carry out extraction treatment to obtain an extraction mixture;

[0159] S3. Filter the extraction mixture obtained in step S2, and then concentrate the filtrate by ultrafiltration membrane to obtain the concentrated liquid of Ilex rotunda Thunb.; the operating pressure of ultrafiltration membrane concentration is 0.4 MPa, and concentrate it to a solid content of 20%;

[0160] S4. Add 3% Tween-80 and 2% glycerol to the Evodia lepta concentrated solution obtained in S3, and then perform spray drying to obtain the Evodia lepta extract. The inlet air temperature for the spray drying process is 170°C, the material temperature is 70°C, and the outlet air temperature is 80°C.

[0161] Example 12

[0162] The extraction method of Evodia lepta medicinal materials in this example includes the following steps:

[0163] S1. Take Evodia lepta medicinal materials, perform comminution treatment, and comminute to a particle size of 65 mesh to obtain medicinal material powder.

[0164] S2. Take a 40% ethanol solution, add citric acid to adjust the pH value to 5.0 as the extraction solvent; take cellulase, pectinase, and neutral protease, and mix them in a mass ratio of 1:2:1 as the enzyme; according to the material-liquid ratio of the medicinal material powder to the extraction solvent of 1:25 and the enzyme addition amount accounting for 2.0% of the mass of the medicinal material powder, add the medicinal material powder, extraction solvent, and enzyme to the extraction unit, control the extraction temperature at 50°C, and the time at 150 min, and perform extraction treatment to obtain an extraction mixture.

[0165] S3. Filter the extraction mixture obtained in step S2, and then concentrate the filtrate by ultrafiltration membrane to obtain the Evodia lepta concentrated solution. The operating pressure for ultrafiltration membrane concentration is 0.4 MPa, and it is concentrated to a solid content of 20%.

[0166] S4. Add 4% Tween-80 to the Evodia lepta concentrated solution obtained in S3, and then perform spray drying to obtain the Evodia lepta extract. The inlet air temperature for the spray drying process is 170°C, the material temperature is 70°C, and the outlet air temperature is 80°C.

[0167] Example 13

[0168] The extraction method of Evodia lepta medicinal materials in this example includes the following steps:

[0169] S1. Take Evodia lepta medicinal materials, perform comminution treatment, and comminute to a particle size of 65 mesh to obtain medicinal material powder.

[0170] S2. Take a 40% ethanol solution, add citric acid to adjust the pH value to 5.0 as the extraction solvent; take cellulase, pectinase, and neutral protease, and mix them in a mass ratio of 1:2:1 as the enzyme; according to the material-liquid ratio of the medicinal material powder to the extraction solvent of 1:25 and the enzyme addition amount accounting for 2.0% of the mass of the medicinal material powder, add the medicinal material powder, extraction solvent, and enzyme to the extraction unit, control the extraction temperature at 50°C, and the time at 150 min, and perform extraction treatment to obtain an extraction mixture.

[0171] S3. Filter the extraction mixture obtained in step S2, and then concentrate the filtrate by ultrafiltration membrane to obtain the Evodia lepta concentrate. The operating pressure for ultrafiltration membrane concentration is 0.4 MPa, and the concentration is carried out until the solid content reaches 20%.

[0172] S4. Add 4% glycerol to the Evodia lepta concentrate obtained in S3, and then carry out spray drying to obtain the Evodia lepta extract. The inlet air temperature for spray drying is 170 °C, the material temperature is 70 °C, and the outlet air temperature is 80 °C.

[0173] Example 14

[0174] The extraction method of Evodia lepta medicinal materials in this example includes the following steps:

[0175] S1. Take Evodia lepta medicinal materials, crush them to a particle size of 65 mesh to obtain medicinal material powder.

[0176] S2. Take a 40% ethanol solution, add citric acid to adjust the pH value to 5.0 as the extraction solvent; take cellulase, pectinase and neutral protease, mix them according to a mass ratio of 1:2:1 as the enzyme; according to the material-liquid ratio of medicinal material powder to extraction solvent of 1:25 and the enzyme addition amount accounting for 2.0% of the mass of the medicinal material powder, add the medicinal material powder, extraction solvent and enzyme into the extraction unit, control the extraction temperature at 50 °C and the time at 150 min, and carry out extraction treatment to obtain the extraction mixture.

[0177] S3. Filter the extraction mixture obtained in step S2, and then concentrate the filtrate by ultrafiltration membrane to obtain the Evodia lepta concentrate. The operating pressure for ultrafiltration membrane concentration is 0.4 MPa, and the concentration is carried out until the solid content reaches 20%.

[0178] S4. Add 2% Tween-80, 1% PVPK30 and 1% glycerol to the Evodia lepta concentrate obtained in S3, and then carry out spray drying to obtain the Evodia lepta extract. The inlet air temperature for spray drying is 170 °C, the material temperature is 70 °C, and the outlet air temperature is 80 °C.

[0179] Similar transfer rate tests were carried out on the Evodia lepta extracts of Examples 6 to 14. At the same time, according to the preparation method of Example 3, Evodia lepta extraction powder was prepared using the Evodia lepta extracts of Examples 6 to 14, and the stability was detected. The detection results are shown in Tables 5 and 6 below.

[0180] Table 5 Transfer rate detection results of Examples 6 - 14

[0181]

[0182] Table 6 Stability detection results of Examples 6 - 14

[0183]

[0184] From the data in Table 5 and in combination with the data in Table 1, it can be seen that when comparing the data of Examples 1 to 14 of the present application, the extraction process of the present application uses a composite enzyme of cellulase, pectinase and neutral protease, and the extraction effect is significantly better than that using a composite enzyme of cellulase and pectinase; in addition, when the present application uses a composite enzyme of cellulase, pectinase and neutral protease and the ratio of the three enzymes is 1:2:1, the extraction effect is optimal. Additionally, for the extraction process of the present application, the pH value of the extraction solvent is controlled at 4.5 - 5.0, the extraction temperature is 48 - 50 °C, and the extraction time is 120 - 150 min, and the extraction effect is relatively optimal.

[0185] From the data in Table 6 and in combination with the data in Table 3, it can be seen that when comparing the data of Examples 1 to 14 of the present application, for the extraction process of the present application, the spray drying uses a composite auxiliary agent, and the stability of the prepared Evodia extract powder is significantly better than that using a single auxiliary agent. In addition, when the composite auxiliary agent selects a mixture of three auxiliary agents, the stability of the prepared Evodia extract powder is significantly better than that using a mixture of two auxiliary agents.

[0186] The above are all preferred embodiments of the present application, and the protection scope of the present application is not limited thereby. Therefore, all equivalent changes made according to the structure, shape, and principle of the present application shall be covered within the protection scope of the present application.

Claims

1. A method for extracting the medicinal material of Jiubiying, characterized in that: The following steps are involved: S1. Take Jiubing medicinal materials, crush them, and obtain medicinal material powder; S2, extracting the medicinal material powder obtained in step S1 by bio-enzyme method using ethanol as extraction solvent and enzyme as extraction auxiliary agent to obtain an extraction mixture; S3, filtering and concentrating the extracted mixed solution obtained in step S2 through an ultrafiltration membrane to obtain a concentrated solution of Jiubing; S4, spray-drying the concentrated solution of the herb obtained in S3 to obtain a herb extract.

2. The method for extracting the herbal medicine of Rhizoma Cibotii according to claim 1, characterized in that: In the step S1, the Jiubiying medicinal material is a processed medicinal material.

3. The method for extracting the herbal medicine of Rhizoma Cibotii according to claim 1, characterized in that: In the step S1, the pulverization process requires pulverizing the Jiubiying medicinal material into a medicinal material powder with a particle size of 24-80 mesh.

4. The method for extracting the herbal medicine of Rhizoma Cibotii according to claim 1, characterized in that: In the step S2, the ethanol is selected to be an ethanol solution with a volume fraction of 20-70%, and the solid-liquid ratio of the medicinal material powder to the extraction solvent is 1:20-30; preferably, the pH value of the ethanol solution is controlled at 4.5-5.5; more preferably, citric acid is used as a regulator to adjust the pH value of the ethanol solution.

5. The method for extracting the herbal medicine of Rhizoma Cibotii according to claim 1, characterized in that: In the step S2, the added amount of the enzyme accounts for 1-4% of the weight of the medicinal material powder.

6. The method for extracting the herbal medicine of Rhizoma Cibotii according to claim 5, characterized in that: In the step S2, the enzyme is a composite enzyme, and the composite enzyme includes cellulase and pectinase; preferably, the mass ratio of the cellulase to the pectinase is 1:

2.

7. The method for extracting the herbal medicine of Rhizoma Cibotii according to claim 6, characterized in that: In the step S2, the complex enzyme includes cellulase, pectinase and neutral protease; preferably, the mass ratio of the cellulase, pectinase and neutral protease is 1:2:

1.

8. The method for extracting the herbal medicine of Rhizoma Cibotii according to claim 1, characterized in that: In step S2, the extraction temperature of the bio-enzymatic extraction treatment is controlled at 45-60° C., and the extraction time is 90-200 min.

9. The method for extracting the herbal medicine of Rhizoma Cibotii according to claim 1, characterized in that: In the step S4, before the spray drying process, an auxiliary agent needs to be added to the schizophylline concentrate, and the auxiliary agent is selected from one or more of Tween-80, sodium alginate, xanthan gum, PEG400, PVPK30 and glycerol; preferably, the amount of the auxiliary agent added accounts for 1-5% of the total mass of the schizophylline concentrate.

10. A Jiubing extract powder, characterized in that: The invention comprises a Herba Scutellariae extract obtained by the Herba Scutellariae extract extraction method according to any one of claims 1 to 9, and auxiliary materials.

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