CN116270805B - Pharmaceutical composition for treating dairy cow mastitis and preparation method thereof - Google Patents
Pharmaceutical composition for treating dairy cow mastitis and preparation method thereof Download PDFInfo
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- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 47
- 208000004396 mastitis Diseases 0.000 title claims abstract description 33
- 235000013365 dairy product Nutrition 0.000 title claims abstract description 21
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- 239000000341 volatile oil Substances 0.000 claims abstract description 43
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- 239000002994 raw material Substances 0.000 claims abstract description 24
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims abstract description 23
- 241000705930 Broussonetia papyrifera Species 0.000 claims abstract description 22
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/282—Artemisia, e.g. wormwood or sagebrush
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/60—Moraceae (Mulberry family), e.g. breadfruit or fig
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0041—Mammary glands, e.g. breasts, udder; Intramammary administration
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- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/14—Drugs for genital or sexual disorders; Contraceptives for lactation disorders, e.g. galactorrhoea
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- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
A pharmaceutical composition for treating cow mastitis and a preparation method thereof are prepared from the following raw materials in percentage by mass: 2-3 parts of argyi leaf essential oil, 5-9 parts of argyi leaf total phenol and 3-6 parts of broussonetia papyrifera leaf total flavone; the preparation method of the pharmaceutical composition comprises the following steps: step 1: extracting mugwort leaf essential oil; step 2: preparing total phenols of folium Artemisiae Argyi; step 3: preparing total flavonoids of broussonetia papyrifera leaves; step 4: the folium artemisiae argyi essential oil, the folium artemisiae argyi total phenol and the folium broussonetiae total flavone are combined according to a certain proportion, and the combination is the pharmaceutical composition for treating the dairy cow mastitis. The pharmaceutical composition for treating the dairy cow mastitis can be directly prepared into corresponding preparations for the dairy cow breast soaking, nipple pouring or smearing on the dairy cow breast, and is used for preventing and treating the dairy cow mastitis.
Description
Technical Field
The invention relates to the technical field of livestock raising, in particular to a pharmaceutical composition for treating dairy cow mastitis and a preparation method thereof.
Background
Bovine mastitis is one of three major diseases in cows. The economic loss caused by cow mastitis worldwide reaches billions of dollars, and the milk production and the quality of milk products are also affected. For a long time, antibiotic medicines are commonly used for treating dairy cow mastitis, but the medicines are used in a large amount, so that the medicines are easy to generate drug resistance and form antibiotic milk, and potential harm is caused to human health.
Disclosure of Invention
The invention aims to provide a pharmaceutical composition for treating cow mastitis and a preparation method thereof.
The above object of the present invention is achieved by the following technical solutions:
a pharmaceutical composition for treating cow mastitis is prepared from the following raw materials in percentage by mass: 2-3 parts of mugwort essential oil, 5-9 parts of mugwort total phenol and 3-6 parts of broussonetia papyrifera total flavone.
Preferably, the pharmaceutical composition for treating cow mastitis is prepared from the following raw materials in percentage by mass: 2 parts of mugwort essential oil, 5 parts of mugwort total phenol and 3 parts of broussonetia papyrifera total flavone.
Preferably, the pharmaceutical composition for treating cow mastitis is prepared from the following raw materials in percentage by mass: 3 parts of argyi leaf essential oil, 9 parts of argyi leaf total phenol and 6 parts of broussonetia papyrifera leaf total flavone.
Preferably, the pharmaceutical composition for treating cow mastitis is prepared from the following raw materials in percentage by mass: 2.5 parts of mugwort essential oil, 7 parts of mugwort total phenol and 4 parts of broussonetia papyrifera total flavone.
A method for preparing a pharmaceutical composition for treating cow mastitis, comprising the following steps:
step 1: extraction of mugwort essential oil: pulverizing fresh folium Artemisiae Argyi, placing into an extraction tank, introducing steam into the extraction tank, extracting volatile oil for 1-2 hr, collecting volatile oil to obtain folium Artemisiae Argyi essential oil with yield (calculated as fresh folium Artemisiae Argyi) of 0.2% -0.5%, and simultaneously obtaining folium Artemisiae Argyi residue for use;
step 2: preparation of total phenols from folium Artemisiae Argyi: adding 3-5 times of water into the mugwort leaf residue, extracting for 2-3 times at 80-90 ℃ for 1-2 hours each time, merging the extracting solutions, concentrating until the mass of raw materials is 0.5-1kg in each liter of concentrated solution, stopping concentrating, centrifuging the concentrated solution at 600-800 r/min, taking supernatant, eluting with water, discarding water eluent until the eluent is nearly colorless, eluting with 30-60% ethanol, collecting 30-60% ethanol eluent, concentrating under reduced pressure until no alcohol exists, and drying (drying under reduced pressure, freeze drying or spray drying) to obtain dry powder, namely mugwort leaf total phenols, wherein the total phenol content is more than 90%, and the total content of cryptochlorogenic acid A, cryptochlorogenic acid B and cryptochlorogenic acid C is more than 20%;
step 3: preparing total flavonoids of broussonetia papyrifera: drying, pulverizing, placing in an extraction tank, adding water solution containing 0.1% NaOH in an amount which is 8-14 times the weight of the raw materials, extracting for 2-4 times at 80-100 ℃ for 1-2h each time, combining the extracting solutions, concentrating under reduced pressure until the mass of the raw materials is 1-1.5 kg/liter of concentrated solution, stopping concentrating, regulating the PH of the concentrated solution to 1-3 by using acid (hydrochloric acid, sulfuric acid, phosphoric acid and the like), standing for 24-48h after precipitation, and vacuum filtering or centrifuging and drying to obtain total flavonoids of the folium Broussonetiae, wherein the yield (calculated by dry folium Broussonetiae) is 3-5%, and the total flavonoids content is 50-70%;
step 4: the mugwort essential oil, mugwort total phenol and broussonetia papyrifera total flavone are combined according to a certain proportion, and the pharmaceutical composition is obtained.
In summary, the beneficial technical effects of the invention are as follows:
the pharmaceutical composition of the dairy cow mastitis can be directly prepared into corresponding preparations for the dairy cow breast soaking, nipple pouring or smearing on the dairy cow breast, and is used for the prevention and treatment of the dairy cow mastitis.
Detailed Description
The present invention will be described in further detail with reference to examples.
Example 1
The invention discloses a pharmaceutical composition for treating cow mastitis, which is prepared from the following raw materials in percentage by mass: 2 parts of mugwort essential oil, 5 parts of mugwort total phenol and 3 parts of broussonetia papyrifera total flavone.
A method for preparing a pharmaceutical composition for treating cow mastitis, comprising the following steps:
step 1: extraction of mugwort essential oil: pulverizing fresh folium Artemisiae Argyi, placing in an extraction tank, introducing steam into the extraction tank, extracting volatile oil for 1 hr, collecting volatile oil to obtain folium Artemisiae Argyi essential oil with yield (calculated as fresh folium Artemisiae Argyi) of 0.2%, and collecting folium Artemisiae Argyi residue;
step 2: preparation of total phenols from folium Artemisiae Argyi: adding 3 times of water into the mugwort leaf residue, extracting for 2 times at 80-90 ℃ for 1.5h each time, combining the extracting solutions, concentrating until the mass of raw materials contained in each liter of concentrated solution is 1kg, stopping concentrating, centrifuging the concentrated solution at 800 revolutions per min, taking supernatant, passing through intermediate macroporous adsorption resin XDA-6, eluting with water, discarding the water eluent until the eluent is nearly colorless, eluting with 40% ethanol, collecting 40% ethanol eluent, concentrating under reduced pressure until no alcohol exists, and freeze-drying to obtain dry powder, namely mugwort leaf total phenols, wherein the yield (calculated by fresh mugwort leaves) is 1.15%, the total phenol content is 94.2%, and the total content of chlorogenic acid A, chlorogenic acid B and chlorogenic acid C is 25.6%;
step 3: preparing total flavonoids of broussonetia papyrifera: drying, pulverizing, placing in an extraction tank, adding 10 times of aqueous solution containing 0.1% NaOH, extracting at 80deg.C for 2 times, 1 hr each time, mixing extractive solutions, concentrating under reduced pressure until the concentrated solution contains 1kg of raw material mass per liter, stopping concentrating, adjusting pH of the concentrated solution to 2 with hydrochloric acid, precipitating, standing for 24 hr, vacuum filtering or centrifuging, and drying to obtain total flavonoids of folium Broussonetiae, wherein the yield (calculated as dry folium Broussonetiae) is 3.8%, and total flavonoids content is 58.6%;
step 4: the mugwort essential oil, mugwort total phenol and broussonetia papyrifera total flavone are combined according to a certain proportion, and the pharmaceutical composition is obtained.
Example 2
The invention discloses a pharmaceutical composition for treating cow mastitis, which is prepared from the following raw materials in percentage by mass: 3 parts of argyi leaf essential oil, 9 parts of argyi leaf total phenol and 6 parts of broussonetia papyrifera leaf total flavone.
A method for preparing a pharmaceutical composition for treating cow mastitis, comprising the following steps:
step 1: extraction of mugwort essential oil: pulverizing fresh folium Artemisiae Argyi, placing in an extraction tank, introducing steam into the extraction tank, extracting volatile oil for 2 hr, collecting volatile oil to obtain folium Artemisiae Argyi essential oil with yield (calculated as fresh folium Artemisiae Argyi) of 0.45%, and collecting folium Artemisiae Argyi residue;
step 2: preparation of total phenols from folium Artemisiae Argyi: adding 5 times of water into the mugwort leaf residue, extracting for 3 times at 80-90 ℃ for 2 hours each time, merging the extracting solutions, concentrating until the mass of raw materials contained in each liter of concentrated solution is 0.8kg, stopping concentrating, centrifuging the concentrated solution at 600 revolutions per minute, taking supernatant, passing through medium-grade macroporous adsorption resin LSA-21, eluting with water, discarding water eluent until the eluent is nearly colorless, eluting with 60% ethanol, collecting 60% ethanol eluent, concentrating under reduced pressure until no alcohol exists, drying under reduced pressure to obtain dry powder of mugwort leaf, namely the mugwort leaf total phenols, wherein the yield (calculated by fresh mugwort leaf) is 1.45%, the total phenols content is 90.5%, and the total content of the chlorogenic acid A, the chlorogenic acid B and the chlorogenic acid C is more than 21.8%;
step 3: preparing total flavonoids of broussonetia papyrifera: drying, pulverizing, placing in an extraction tank, adding water solution containing 0.1% NaOH in an amount which is 14 times the weight of the raw materials, extracting at 95 ℃ for 3 times, 1.5 hours each time, combining the extracting solutions, concentrating under reduced pressure until the mass of 1kg of raw materials is contained in each liter of concentrated solution, stopping concentrating, regulating the PH of the concentrated solution to 1.5 by sulfuric acid, standing for 48 hours, and vacuum filtering or centrifugally separating and drying to obtain total flavonoids of the folium Broussonetiae, wherein the yield (calculated by dry folium Broussonetiae) is 4.6%, and the total flavonoids content is 52.8%;
step 4: the total flavonoids of broussonetia papyrifera, the essential oils of mugwort leaves and the total flavonoids of mugwort leaves are combined according to a certain proportion, and the pharmaceutical composition of the invention is obtained.
Example 3
The invention discloses a pharmaceutical composition for treating cow mastitis, which is prepared from the following raw materials in percentage by mass: 2.5 parts of mugwort essential oil, 7 parts of mugwort total phenol and 4 parts of broussonetia papyrifera total flavone.
A method for preparing a pharmaceutical composition for treating cow mastitis, comprising the following steps:
step 1: extraction of mugwort essential oil: pulverizing fresh folium Artemisiae Argyi, placing in an extraction tank, introducing steam into the extraction tank, extracting volatile oil for 1.5 hr, collecting volatile oil to obtain folium Artemisiae Argyi essential oil with yield (calculated as fresh folium Artemisiae Argyi) of 0.4%, and simultaneously obtaining folium Artemisiae Argyi residue;
step 2: preparation of total phenols from folium Artemisiae Argyi: adding 4 times of water into the mugwort leaf residue, extracting for 2 times at 80 ℃ for 1h each time, merging the extracting solutions, concentrating until the mass of raw materials is 0.5kg in each liter of concentrated solution, stopping concentrating, centrifuging the concentrated solution at 800 revolutions per min, taking supernatant, passing through medium-grade macroporous adsorption resin LX-69B, eluting with water, discarding water eluent until the eluent is nearly colorless, eluting with 45% ethanol, collecting 45% ethanol eluent, concentrating under reduced pressure until no alcohol exists, and spray drying to obtain dry powder, namely mugwort leaf total phenols, wherein the yield (calculated by fresh mugwort leaves) is 1.26%, the total phenol content is 92.3%, and the sum of the contents of chlorogenic acid A, chlorogenic acid B and chlorogenic acid C is more than 20%;
step 3: preparing total flavonoids of broussonetia papyrifera: drying folium Broussonetiae, pulverizing, placing in an extraction tank, adding 12 times of aqueous solution containing 0.1% NaOH, extracting at 80deg.C for 3 times, 1.5 hr each time, mixing extractive solutions, concentrating under reduced pressure until the concentration liquid contains 1kg of raw material per liter, regulating pH of the concentrated liquid to 2 with acid (hydrochloric acid, sulfuric acid, phosphoric acid, etc.), standing for 36 hr, vacuum filtering or centrifuging, and drying to obtain total flavonoids of folium Broussonetiae, and collecting the total flavonoids in yield (calculated as dry folium Broussonetiae)
4.3% of total flavonoids content of 56.5%;
step 4: the mugwort essential oil, mugwort total phenol and broussonetia papyrifera total flavone are combined according to a certain proportion, and the pharmaceutical composition is obtained.
The following experiments were performed in conjunction with the above examples 1-3
Test 1: effect of pharmaceutical composition for inhibiting LPS-induced bovine mammary epithelial cell MAC-T inflammatory injury
Staphylococcus aureus, streptococcus and escherichia coli are main pathogenic bacteria of cow mastitis, pathogenic microorganisms invade mammary tissue through nipple ducts to cause mammary gland to be infected, so that the mammary gland area is dilated, blood flow is slow, capillary permeability is increased, local blood flow disorder is caused, and various inflammatory mediators such as TNF-alpha, IL-6, IL-1 beta and the like in blood are increased.
According to the experiment, a bovine mammary gland epithelial cell inflammation model is induced by Lipopolysaccharide (LPS), the folium artemisiae argyi essential oil, the folium artemisiae argyi total phenol, the folium broussonetiae total flavone and the pharmaceutical composition prepared in the examples 1-3 are respectively used as test samples, the cell activity of the bovine mammary gland epithelial cell inflammation model and the contents of inflammatory factors TNF-alpha, IL-6 and IL-1 beta in cells are detected, and the protection effect of the folium artemisiae argyi essential oil, the folium artemisiae argyi total phenol, the folium broussonetiae total flavone and the pharmaceutical composition disclosed by the invention on cell inflammation injury is examined.
Results: after LPS acts on bovine mammary gland epithelial cells, the cell activity is obviously reduced (P is less than 0.05), and the secretion of inflammatory factors TNF-alpha, IL-6 and IL-1 beta in the cells is increased (P is less than 0.05); after the pharmaceutical composition prepared by the examples 1-3 is used for inflammatory cells, the cell activity is obviously increased (P is less than 0.05), the content of inflammatory factors TNF-alpha, IL-6 and IL-1 beta in the cells is obviously reduced (P is less than 0.05), wherein after the pharmaceutical composition prepared by the examples 1-3 is used for inflammatory cells, the cell activity is obviously better than that of the essential oil of the folium artemisiae argyi, the total flavonoids of the folium artemisiae argyi (P is less than 0.05), the content of the inflammatory factors TNF-alpha, IL-6 and IL-1 beta in the cells is also obviously lower than that of any one component of the essential oil of the folium artemisiae argyi, the total flavonoids of the folium artemisiae argyi (P is less than 0.05), and the content of the inflammatory factors TNF-alpha, IL-6 and IL-1 beta in the cells is also obviously lower than that of the three components are singly used (P is less than 0.05), so that the pharmaceutical composition can weaken the inflammatory injury of cells induced by LPS by inhibiting the production of TNF-alpha, IL-6 and IL-1 beta in mammary epithelial cells.
Table 1. Effect of the pharmaceutical composition of the invention on LPS-induced bovine mammary epithelial cell inflammation model (n=9)
Note that: the same column of data, different letters indicate significant differences (P < 0.05), and the same letters indicate insignificant differences (P > 0.05).
Test 2 clinical trial of the pharmaceutical composition on recessive mastitis in cows
30 cows of Chinese Holstein lactation period which are fed in a certain dairy farm and are diagnosed with recessive mastitis are selected, and the milk of the sick cows is detected to be weak positive (the number of somatic cells is more than 50 ten thousand per ml) through SMT, so that the milk yield is reduced. The sick cows are randomly divided into 3 groups, namely a treatment group, a non-treatment group and a western medicine control group, 10 cows are fed separately, and the feeding conditions are identical. The treatment group uses the breast perfusate prepared by the pharmaceutical composition of the invention to carry out breast infusion treatment, the western medicine control group uses the control medicine to carry out breast infusion treatment, 100ml each time is carried out, the treatment course is 7 days, and the control group does not adopt any medicine treatment. After the end of the administration, the milk samples were taken for the detection of the somatic cell count (somatic cell counter) and the inflammatory factors TNF-alpha, IL-6 and IL-1β in milk (ELISA kit detection), and the results are shown in Table 2.
The preparation method of the breast perfusate prepared by the pharmaceutical composition comprises the following steps: the pharmaceutical composition in example 3 was prepared by mixing 5g of the pharmaceutical composition, 2g of carbomer, 1ml of Tween-80, 2ml of ethanol and 90ml of distilled water, shearing the mixture with a high-speed shearing machine until the solution became transparent, filtering, filling and sterilizing.
Control drug: 100mg of ribavirin, 5mg of dexamethasone and 2g of ciprofloxacin are mixed and dissolved in 100mL of physiological saline.
TABLE 2 variation of somatic cell count and inflammatory factors TNF- α, IL-6 and IL-1β in milk of sick cattle (n=10)
Note that: the same column of data, different letters indicate significant differences (P < 0.05), and the same letters indicate insignificant differences (P > 0.05).
Results: compared with untreated groups, the breast perfusion agent prepared from the pharmaceutical composition can obviously reduce the quantity of somatic cells in milk of cows (P is less than 0.05), and simultaneously, the content of inflammatory factors TNF-alpha, IL-6 and IL-1 beta in whey is also obviously reduced (P is less than 0.05).
The medicine composition has good application effect on the increase of somatic cells in milk and the increase of inflammatory factors caused by recessive mastitis of cows.
The pharmaceutical composition of the dairy cow mastitis can be directly prepared into corresponding preparations for the dairy cow breast soaking, nipple pouring or smearing on the dairy cow breast, and is used for the prevention and treatment of the dairy cow mastitis.
The embodiments of the present invention are all preferred embodiments of the present invention, and are not intended to limit the scope of the present invention in this way, therefore: all equivalent changes in structure, shape and principle of the invention should be covered in the scope of protection of the invention.
Claims (4)
1. A pharmaceutical composition for treating cow mastitis, comprising: the material is prepared from the following raw materials in percentage by mass: 2-3 parts of argyi leaf essential oil, 5-9 parts of argyi leaf total phenol and 3-6 parts of broussonetia papyrifera leaf total flavone;
the preparation method of the pharmaceutical composition comprises the following steps:
step 1: extraction of mugwort essential oil: pulverizing fresh folium Artemisiae Argyi, placing into an extraction tank, introducing steam into the extraction tank, extracting volatile oil for 1-2 hr, collecting volatile oil to obtain folium Artemisiae Argyi essential oil with yield of 0.2% -0.5%, and simultaneously obtaining folium Artemisiae Argyi residue for use;
step 2: preparation of total phenols from folium Artemisiae Argyi: adding 3-5 times of water into the mugwort leaf residue, extracting for 2-3 times at 80-90 ℃ for 1-2 hours each time, merging the extracting solutions, concentrating until the mass of raw materials is 0.5-1kg in each liter of concentrated solution, stopping concentrating, centrifuging the concentrated solution at 600-800 r/min, taking supernatant, passing through a medium-polarity macroporous adsorption resin, eluting with water, discarding the water eluent until the eluent is nearly colorless, eluting with 30-60% ethanol, collecting 30-60% ethanol eluent, concentrating under reduced pressure until no alcohol exists, and drying to obtain dry powder, namely mugwort leaf total phenols, wherein the total phenol content is more than 90%, and the total content of chlorogenic acid A, chlorogenic acid B and chlorogenic acid C is more than 20%;
step 3: preparing total flavonoids of broussonetia papyrifera: drying, pulverizing, placing in an extraction tank, adding 8-14 times of aqueous solution containing 0.1% NaOH, extracting at 80-100deg.C for 2-4 times, each time for 1-2 hr, mixing extractive solutions, concentrating under reduced pressure until each liter of concentrated solution contains 1-1.5kg of raw material mass, stopping concentrating, regulating pH of concentrated solution to 1-3 with acid, standing for 24-48 hr, vacuum filtering or centrifuging, and drying to obtain total flavonoids of folium Broussonetiae, wherein the total flavonoids content is 50-70%;
step 4: the folium artemisiae argyi essential oil, the folium artemisiae argyi total phenol and the folium broussonetiae total flavone are combined according to a certain proportion to obtain the pharmaceutical composition.
2. A pharmaceutical composition for bovine mastitis according to claim 1, wherein: the pharmaceutical composition for treating the dairy cow mastitis is prepared from the following raw materials in percentage by mass: 2 parts of mugwort essential oil, 5 parts of mugwort total phenol and 3 parts of broussonetia papyrifera total flavone.
3. A pharmaceutical composition for bovine mastitis according to claim 1, wherein: the pharmaceutical composition for treating the dairy cow mastitis is prepared from the following raw materials in percentage by mass: 3 parts of argyi leaf essential oil, 9 parts of argyi leaf total phenol and 6 parts of broussonetia papyrifera leaf total flavone.
4. A pharmaceutical composition for bovine mastitis according to claim 1, wherein: the pharmaceutical composition for treating the dairy cow mastitis is prepared from the following raw materials in percentage by mass: 2.5 parts of mugwort essential oil, 7 parts of mugwort total phenol and 4 parts of broussonetia papyrifera total flavone.
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