CN102772520B - Traditional Chinese medicine composition for preventing or treating bovine mastitis and preparation method of composition - Google Patents
Traditional Chinese medicine composition for preventing or treating bovine mastitis and preparation method of composition Download PDFInfo
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Abstract
The invention discloses a traditional Chinese medicine composition for preventing or treating bovine mastitis and a preparation method of the composition. The traditional Chinese medicine composition comprises the following bulk drugs in parts by weight: 20-40 parts of safflower, 3-18 parts of sanguisorba officinalis, 3-18 parts of gallnut, 3-18 parts of myrobalan, 10-25 parts of rhizoma coptidis and 3-18 parts of radix bupleuri. The traditional Chinese medicine composition has the pharmacological efficacies of clearing away heat and toxic materials, diminishing swelling, removing stasis and enhancing immunity of a human body, can be prepared into transdermal agents, is convenient to use, has no toxic or side effect and stress response, is safe and effective, has no drug resistance, and is low in cost. The components of the composition can directly and rapidly reach the focus. Proven by clinic effect viewing test results, the traditional Chinese medicine composition disclosed by the invention is rapid in taking effect on bovine mastitis, definite in curative effect, and has no toxic or side effect, and is good in safety.
Description
Technical field
The present invention relates to a kind of medicine for the treatment of cow breast disease, relate in particular to the Chinese medicine composition of a kind of prevention or treatment mammitis of cow, the invention still further relates to the various pharmaceutical preparatioies that are prepared from by this Chinese medicine composition, the invention belongs to the treatment by Chinese herbs field of mammitis of cow.
Background technology
Mammitis of cow be dairy sickness rate the highest, cause one of disease that economic loss is the most serious.Have or not naked eyes visible change according to breast and milk, mammitis of cow is divided into two types of clinic mastitis and latent mammitises.Latent mammitis does not have a macroscopic variation in appearance milch cow and breast thereof, suffer from the sick cattle of latent mammitis, in its milk, protein content declines, bacterial content raises, milk composition changes, cause the quality of sweet milk to decline, reduce its nutritive value, in addition, the residual antibiotic of the pathogenic microorganism in milk and western medicine causes a large amount of milk to be abandoned.
At present, China's bovine subclinical mastitis common antibiotics treatment, majority takes the mode of drug combination to give broad-spectrum antibiotic, causes pathogen to strengthen the drug resistance of antibacterials, usually occur that treatment cycle extends, dosage increases, serious appearance is failed to respond to any medical treatment and milch cow is eliminated situation.The increase of administering drug combinations and dosage causes antibiotics residue problem day by day serious.
Chinese medicine has effect of heat-clearing and toxic substances removing, dispersing swelling and dissipating binds, and Chinese medicine also can strengthen animal body immunologic function simultaneously, and is difficult for producing drug resistance.So treatment by Chinese herbs mastitis is development trend from now on.Chinese medicine is at present generally by oral administration or intravenous administration, for mastitis there is the shortcoming that drug effect is low, therapeutic effect is poor in treatment, if can directly can greatly improve in breast topical effect, give so study external part for mastitis the optimum selection that pure Chinese medicine preparation capable of permeating skin is treatment.
Summary of the invention
Technical problem underlying to be solved by this invention is to provide the Chinese medicine composition of a kind of evident in efficacy, prevention of having no side effect or treatment mammitis of cow;
Technical problem underlying to be solved by this invention is achieved through the following technical solutions:
A Chinese medicine composition for prevention or treatment mammitis of cow, comprises that the crude drug of following weight portion is made: Flos Carthami 20-40 part, Radix Sanguisorbae 3-18 part, Galla Chinensis 3-18 part, Fructus Chebulae 3-18 part, Rhizoma Coptidis 10-25 part and Radix Bupleuri 3-18 part; The present invention finds by a large amount of pharmacodynamics tests, while containing the Herba Violae of 5-20 weight portion, can play better prevention or therapeutical effect for mammitis of cow in this Chinese medicine composition.
The weight portion of each crude drug is preferably: Flos Carthami 20-25 part, Radix Sanguisorbae 5-10 part, Galla Chinensis 5-10 part, Fructus Chebulae 5-10 part, Rhizoma Coptidis 15-20 part, Radix Bupleuri 5-10 part; Wherein, in the time of Herba Violae that this Chinese medicine composition contains 10-15 weight portion, can play better prevention or therapeutical effect for mammitis of cow.
Preferred, the weight portion of each crude drug is: 22 parts, Flos Carthami, 8 parts of Radix Sanguisorbaes, 8 parts of Galla Chinensiss, 8 parts of Fructus Chebulaes, 18 parts of Rhizoma Coptidis, 8 parts of Radix Bupleuri; Wherein, in the time of Herba Violae that this Chinese medicine composition contains 12 weight portions, can play better prevention or therapeutical effect for mammitis of cow.
Chinese medicine composition of the present invention is under Chinese medical theory instructs, achievement in research according to modern Chinese medicine pharmacy to mammitis of cow treatment, in conjunction with inventor's clinical practice experience for many years, filter out the crude drug of eliminating stasis to stop pain, heat-clearing and toxic substances removing, removing heat from blood detumescence, according to theory of Chinese medical science prescription, form according to certain weight prescription.
Flos Carthami: formal name used at school Carthamus tinctorius L. is the tubular corolla of feverfew Flos Carthami Carthamus tinctorius L..Warm in nature, acrid in the mouth; Invigorate blood circulation latus rectum, eliminating stasis to stop pain.For amenorrhea, dysmenorrhea, lochia, lump in the abdomen mass in the abdomen, traumatic injury.
Radix Sanguisorbae: formal name used at school: Radix Sanguisorbae is the plant of Rosaceae burnet.Property of medicine picric acid; Cold; Nontoxic, return liver, lung, kidney, large intestine channel.Effect: cooling blood for hemostasis, heat-clearing and toxic substances removing, detumescence sore.
Galla Chinensis: latin name: Galla chinensi, have another name called hundred worm storehouses, Mass Galla chinesis et camelliae Fermentata, gall, for the Chinese Nut-gall aphid of Homoptera aphid section or the female worm of melaphis peitan Tsai et Tang parasitize Anacardiaceae plant " Fructus rhois chinensis " and belong to tender leaf or the petiole of other plant together, stab and generate a kind of cryptomere consor thing insect gall, through dry doubly dry after gained.Meridian distribution of property and flavor: sour in the mouth is puckery, cold in nature.Return lung, large intestine, kidney channel.Effect: astringe the lung; Hidroschesis; Astringing intestine to stop diarrhea; Controlling nocturnal emission with astringent drugs; Hemostasis; Removing toxic substances.
Fructus Chebulae: another name: chebule, scold multitude, scold pears, son with the wind; English name: Medicine Terminalia Fruit, Fruit of medicine Terminalia.Bitter in the mouth; Acid; Puckery; Property is put down.Effect: astringe the lung; Astringing intestine to stop diarrhea; The therapeutic method to keep the adverse QI flowing downwards; Sore-throat relieving.
Rhizoma Coptidis: be the dry rhizome of cohosh Rhizoma Coptidis (Coptis chinensis Franch.), Coptis deltoidea C.Y.Cheng et Hsiao (Coptis deltoidea C.Y.Cheng et Hsiao) and Coptis Teeta Wall (Coptisteeta Wall.).Meridian distribution of property and flavor: hardship, cold, nontoxic.GUIXIN, spleen, stomach, liver, gallbladder, large intestine channel.Function cures mainly: heat clearing and damp drying, eliminating fire and detoxication.
Radix Bupleuri: cold nature, bitter in the mouth, pungent, return Liver Channel, gallbladder meridian.Function: expelling pathogenic factors from the exterior expels the heat-evil, dispersing the stagnated live-QI to relieve the stagnation of QI, elevate a turnable ladder yang-energy.
Herba Violae: formal name used at school: Purplef lower Violet.Another name: Herba violae patrinii, light lobe common violet.Property bitter, acrid, cold, nontoxic; GUIXIN, Liver Channel.Function cures mainly: heat-clearing and toxic substances removing, removing heat from blood detumescence.Traumatic injury, carbuncle etc. are controlled in external application.
The present invention's various crude drug used all can be bought and obtain from a medical store, and its specification meets country or industry standard.
Chinese medicine composition of the present invention can be prepared into any acceptable suitable formulations clinically by this area conventional formulation method, for example, can such as, to adding in the active component of medicine of the present invention required various adjuvants (disintegrating agent, lubricant, adhesive, transdermal enhancer etc.) while preparing different dosage form to be prepared into any suitable clinical preparation with conventional method of Chinese medicinal, for example, can be various external preparation, oral formulations or ejection preparation; Wherein, described external preparation is preferably transdermal agent, unguentum, cataplasma etc., most preferably is transdermal agent; Described oral formulations is preferably powder or decoction etc.
Preferably, a kind of method of preparing above-mentioned Chinese medicine composition, the method comprises the following steps:
(1) take each crude drug by described weight portion;
(2) each crude drug is obtained to Chinese medicine extraction liquid with water extraction;
(3) Chinese medicine extraction liquid and triethanolamine are mixed to get to water, by adding again transdermal enhancer after water and oil phase mixing, obtain transdermal agent.
Wherein, the preparation method of described Chinese medicine extraction liquid can be prepared with reference to following methods:
(a) each crude drug is obtained to extracting solution with water extraction; By concentrated extracting solution, filtration;
(b) filtrate adds ethanol, and alcohol deposit fluid is centrifugal, gets supernatant; Supernatant adds ethanol to carry out precipitate with ethanol after vaporing away ethanol again, and alcohol deposit fluid is filtered, and filtered solution vapors away ethanol and adds distilled water again, with filtering with microporous membrane, obtains Chinese medicine extraction liquid.
Wherein, described transdermal enhancer can be azone, 1,2-PD, dimethyl sulfoxide, acetylated monoglyceride etc.
Described oil phase comprises stearic acid, glyceryl monostearate, vaseline or paraffin etc.;
The present invention also provides a kind of method of preparing above-mentioned Chinese medicine composition, and the method comprises the following steps:
(1) take each crude drug by described weight portion;
(2) by each crude drug water or ethanol extraction, filter, add the required adjuvant of preparations shaping or excipient, be prepared into decoction or powder.
Chinese medicine composition of the present invention has effect of heat-clearing and toxic substances removing, dispersing swelling and dissipating binds, enhancing human body immunity function, can be prepared into transdermal agent, ingredient can arrive disease sites directly, rapidly, easy to use, have no side effect and stress, safe and effective, without Drug resistance, with low cost.The confirmation of clinical observation on the therapeutic effect result of the test, Chinese medicine composition of the present invention, for mammitis of cow instant effect, determined curative effect, has no side effect, and safety is good.
Using method and consumption: the dosage of Chinese medicine composition of the present invention depends on concrete dosage form.As guidance: for transdermal agent; its using method is as follows: before medication, clean the ill newborn district of disease cattle with warm water; with after clean towel; the transdermal agent 20-30mL preparing with Chinese medicine composition of the present invention is smeared in each newborn district; smear generally massage, 2 times/d on one side; 15-20min/ time, 5d is 1 course for the treatment of.
The specific embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage and disadvantage of the present invention will be more clear along with description.But these embodiment are only exemplary, scope of the present invention are not formed to any restriction.It will be understood by those skilled in the art that lower without departing from the spirit and scope of the present invention and can the details of technical solution of the present invention and form be modified or be replaced, but these modifications and replacement all fall within the scope of protection of the present invention.
The preparation of embodiment 1 transdermal agent
1, by described weight take each crude drug (unit: g): Flos Carthami 22, Radix Sanguisorbae 8, Galla Chinensis 8, Fructus Chebulae 8, Rhizoma Coptidis 18, Radix Bupleuri 8;
2, each crude drug is mixed, by the mixture 1000mL that adds water, soaks after 4 hours, decocts 1 hour, collect decoction liquor, then add 500mL water to decoct for the second time 30 minutes, collection decoction liquor, then add 500mL water to decoct for the third time 30 minutes, collection decoction liquor, when each decoction, the backflow distilling out is reclaimed in condensation, and above-mentioned three decoction liquor and backflow are merged, and 70 ℃ of water-baths are concentrated into 150mL and filter to obtain filtrate, it is 95% ethanol that filtrate adds concentration, make concentration of alcohol reach 60%, after fully stirring, obtain alcohol deposit fluid, alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 24 hours, centrifugal 5 minutes of 3000r/min, get 70 ℃ of water-baths of supernatant, volatilization goes to add concentration 95% ethanol after ethanol again, make concentration of alcohol reach 80%, again alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 24 hours, filter paper filters to obtain filtered solution, 70 ℃ of water-baths of filtered solution, ethanol is removed in volatilization, filtered solution adds distilled water to 100mL, making solution contain crude drug amount is 1g/mL, obtain filtrate with 0.25 micron of filtering with microporous membrane, filtrate steam sterilization 30min at 121 ℃ obtains Chinese medicine extraction liquid,
The Chinese medicine extraction liquid preparing and triethanolamine are mixed to get to water; After water is mixed with oil phase (stearic acid, glyceryl monostearate, vaseline or paraffin), add again transdermal agent (azone, 1,2-PD, dimethyl sulfoxide or acetylated monoglyceride), obtain Chinese medicine transdermal agent.
The preparation of embodiment 2 transdermal agents
1, by described weight take each crude drug (unit: g): Flos Carthami 20, Radix Sanguisorbae 3, Galla Chinensis 3, Fructus Chebulae 3, Rhizoma Coptidis 10, Radix Bupleuri 3;
2, each crude drug is mixed, by the mixture 1000mL that adds water, soaks after 2 hours, decocts 1 hour, collect decoction liquor, then add 500mL water to decoct for the second time 30 minutes, collection decoction liquor, then add 500mL water to decoct for the third time 30 minutes, collection decoction liquor, when each decoction, the backflow distilling out is reclaimed in condensation, and above-mentioned three decoction liquor and backflow are merged, and 70 ℃ of water-baths are concentrated into 150mL and filter to obtain filtrate, it is 95% ethanol that filtrate adds concentration, make concentration of alcohol reach 60%, after fully stirring, obtain alcohol deposit fluid, alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 24 hours, centrifugal 5 minutes of 3000r/min, get 70 ℃ of water-baths of supernatant, volatilization goes to add concentration 95% ethanol after ethanol again, make concentration of alcohol reach 80%, again alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 24 hours, filter paper filters to obtain filtered solution, 70 ℃ of water-baths of filtered solution, ethanol is removed in volatilization, filtered solution adds distilled water to 100mL, making solution contain crude drug amount is 1g/mL, obtain filtrate with 0.25 micron of filtering with microporous membrane, filtrate steam sterilization 30min at 121 ℃ obtains Chinese medicine extraction liquid,
The Chinese medicine extraction liquid preparing and triethanolamine are mixed to get to water; After water is mixed with oil phase (stearic acid, glyceryl monostearate, vaseline or paraffin), add again transdermal agent (azone, 1,2-PD, dimethyl sulfoxide or acetylated monoglyceride), obtain Chinese medicine transdermal agent.
The preparation of embodiment 3 transdermal agents
1, by described weight take each crude drug (unit: g): Flos Carthami 40, Radix Sanguisorbae 18, Galla Chinensis 18, Fructus Chebulae 18, Rhizoma Coptidis 25, Radix Bupleuri 18;
2, each crude drug is mixed, by the mixture 1000mL that adds water, soaks after 5 hours, decocts 1.5 hours, collect decoction liquor, then add 500mL water to decoct for the second time 40 minutes, collection decoction liquor, then add 500mL water to decoct for the third time 40 minutes, collection decoction liquor, when each decoction, the backflow distilling out is reclaimed in condensation, and above-mentioned three decoction liquor and backflow are merged, and 70 ℃ of water-baths are concentrated into 150mL and filter to obtain filtrate, it is 95% ethanol that filtrate adds concentration, make concentration of alcohol reach 60%, after fully stirring, obtain alcohol deposit fluid, alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 36 hours, centrifugal 5 minutes of 3000r/min, get 70 ℃ of water-baths of supernatant, volatilization goes to add concentration 95% ethanol after ethanol again, make concentration of alcohol reach 80%, again alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 36 hours, filter paper filters to obtain filtered solution, 70 ℃ of water-baths of filtered solution, ethanol is removed in volatilization, filtered solution adds distilled water to 100mL, making solution contain crude drug amount is 1g/mL, obtain filtrate with 0.25 micron of filtering with microporous membrane, filtrate steam sterilization 30mi n at 121 ℃ obtains Chinese medicine extraction liquid,
The Chinese medicine extraction liquid preparing and triethanolamine are mixed to get to water; After water is mixed with oil phase (stearic acid, glyceryl monostearate, vaseline or paraffin), add again transdermal agent (azone, 1,2-PD, dimethyl sulfoxide or acetylated monoglyceride), obtain Chinese medicine transdermal agent.
The preparation of embodiment 4 transdermal agents
1, by described weight take each crude drug (unit: g): Flos Carthami 20, Radix Sanguisorbae 5, Galla Chinensis 5, Fructus Chebulae 5, Rhizoma Coptidis 15, Radix Bupleuri 5, Herba Violae 10;
2, each crude drug is mixed, by the mixture 1000mL that adds water, soaks after 3 hours, decocts 1 hour, collect decoction liquor, then add 500mL water to decoct for the second time 30 minutes, collection decoction liquor, then add 500mL water to decoct for the third time 30 minutes, collection decoction liquor, when each decoction, the backflow distilling out is reclaimed in condensation, and above-mentioned three decoction liquor and backflow are merged, and 70 ℃ of water-baths are concentrated into 150mL and filter to obtain filtrate, it is 95% ethanol that filtrate adds concentration, make concentration of alcohol reach 60%, after fully stirring, obtain alcohol deposit fluid, alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 24 hours, centrifugal 5 minutes of 3000r/mi n, get 70 ℃ of water-baths of supernatant, volatilization goes to add concentration 95% ethanol after ethanol again, make concentration of alcohol reach 80%, again alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 24 hours, filter paper filters to obtain filtered solution, 70 ℃ of water-baths of filtered solution, ethanol is removed in volatilization, filtered solution adds distilled water to 100mL, making solution contain crude drug amount is 1g/mL, obtain filtrate with 0.25 micron of filtering with microporous membrane, filtrate steam sterilization 30min at 121 ℃ obtains Chinese medicine extraction liquid,
The Chinese medicine extraction liquid preparing and triethanolamine are mixed to get to water; After water is mixed with oil phase (stearic acid, glyceryl monostearate, vaseline or paraffin), add again transdermal agent (azone, 1,2-PD, dimethyl sulfoxide or acetylated monoglyceride), obtain Chinese medicine transdermal agent.
The preparation of embodiment 5 transdermal agents
1, by described weight take each crude drug (unit: g): Flos Carthami 25, Radix Sanguisorbae 10, Galla Chinensis 10, Fructus Chebulae 10, Rhizoma Coptidis 20, Radix Bupleuri 10, Herba Violae 15;
2, each crude drug is mixed, by the mixture 1000mL that adds water, soak after 4h, decocts 1 hour, collect decoction liquor, then add 500mL water to decoct for the second time 40min, collect decoction liquor, then add 500mL water to decoct for the third time 40min, collection decoction liquor, when each decoction, the backflow distilling out is reclaimed in condensation, and above-mentioned three decoction liquor and backflow are merged, and 70 ℃ of water-baths are concentrated into 150mL and filter to obtain filtrate, it is 95% ethanol that filtrate adds concentration, make concentration of alcohol reach 60%, after fully stirring, obtain alcohol deposit fluid, alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 24h, the centrifugal 5min of 3000r/min, get 70 ℃ of water-baths of supernatant, volatilization goes to add concentration 95% ethanol after ethanol again, make concentration of alcohol reach 80%, again alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 24h, filter paper filters to obtain filtered solution, 70 ℃ of water-baths of filtered solution, ethanol is removed in volatilization, filtered solution adds distilled water to 100mL, making solution contain crude drug amount is 1g/mL, obtain filtrate with 0.25 micron of filtering with microporous membrane, filtrate steam sterilization 30min at 121 ℃ obtains Chinese medicine extraction liquid,
The Chinese medicine extraction liquid preparing and triethanolamine are mixed to get to water; After water is mixed with oil phase (stearic acid, glyceryl monostearate, vaseline or paraffin), add again transdermal agent (azone, 1,2-PD, dimethyl sulfoxide or acetylated monoglyceride), obtain Chinese medicine transdermal agent.
The preparation of embodiment 6 transdermal agents
1, by described weight take each crude drug (unit: g): Flos Carthami 22, Radix Sanguisorbae 8, Galla Chinensis 8, Fructus Chebulae 8, Rhizoma Coptidis 18, Radix Bupleuri 8, Herba Violae 12;
2, each crude drug is mixed, by the mixture 1000mL that adds water, soaks after 4 hours, decocts 1 hour, collect decoction liquor, then add 500mL water to decoct for the second time 30 minutes, collection decoction liquor, then add 500mL water to decoct for the third time 30 minutes, collection decoction liquor, when each decoction, the backflow distilling out is reclaimed in condensation, and above-mentioned three decoction liquor and backflow are merged, and 70 ℃ of water-baths are concentrated into 150mL and filter to obtain filtrate, it is 95% ethanol that filtrate adds concentration, make concentration of alcohol reach 60%, after fully stirring, obtain alcohol deposit fluid, alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 24 hours, centrifugal 5 minutes of 3000r/min, get 70 ℃ of water-baths of supernatant, volatilization goes to add concentration 95% ethanol after ethanol again, make concentration of alcohol reach 80%, again alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 24 hours, filter paper filters to obtain filtered solution, 70 ℃ of water-baths of filtered solution, ethanol is removed in volatilization, filtered solution adds distilled water to 100mL, making solution contain crude drug amount is 1g/mL, obtain filtrate with 0.25 micron of filtering with microporous membrane, filtrate steam sterilization 30min at 121 ℃ obtains Chinese medicine extraction liquid,
The Chinese medicine extraction liquid preparing and triethanolamine are mixed to get to water; After water is mixed with oil phase (stearic acid, glyceryl monostearate, vaseline or paraffin), add again transdermal agent (azone, 1,2-PD, dimethyl sulfoxide or acetylated monoglyceride), obtain Chinese medicine transdermal agent.
The preparation of embodiment 7 transdermal agents
1, by described weight take each crude drug (unit: g): Flos Carthami 20, Radix Sanguisorbae 3, Galla Chinensis 3, Fructus Chebulae 3, Rhizoma Coptidis 10, Radix Bupleuri 3, Herba Violae 5;
2, each crude drug is mixed, by the mixture 1000mL that adds water, soaks after 2 hours, decocts 1 hour, collect decoction liquor, then add 500mL water to decoct for the second time 30 minutes, collection decoction liquor, then add 500mL water to decoct for the third time 30 minutes, collection decoction liquor, when each decoction, the backflow distilling out is reclaimed in condensation, and above-mentioned three decoction liquor and backflow are merged, and 70 ℃ of water-baths are concentrated into 150mL and filter to obtain filtrate, it is 95% ethanol that filtrate adds concentration, make concentration of alcohol reach 60%, after fully stirring, obtain alcohol deposit fluid, alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 24 hours, centrifugal 5 minutes of 3000r/min, get 70 ℃ of water-baths of supernatant, volatilization goes to add concentration 95% ethanol after ethanol again, make concentration of alcohol reach 80%, again alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 24 hours, filter paper filters to obtain filtered solution, 70 ℃ of water-baths of filtered solution, ethanol is removed in volatilization, filtered solution adds distilled water to 100mL, making solution contain crude drug amount is 1g/mL, obtain filtrate with 0.25 micron of filtering with microporous membrane, filtrate steam sterilization 30min at 121 ℃ obtains Chinese medicine extraction liquid,
The Chinese medicine extraction liquid preparing and triethanolamine are mixed to get to water; After water is mixed with oil phase (stearic acid, glyceryl monostearate, vaseline or paraffin), add again transdermal agent (azone, 1,2-PD, dimethyl sulfoxide or acetylated monoglyceride), obtain Chinese medicine transdermal agent.
The preparation of embodiment 8 transdermal agents
1, by described weight take each crude drug (unit: g): Flos Carthami 40, Radix Sanguisorbae 18, Galla Chinensis 18, Fructus Chebulae 18, Rhizoma Coptidis 25, Radix Bupleuri 18, Herba Violae 20;
2, each crude drug is mixed, by the mixture 1000mL that adds water, soaks after 5 hours, decocts 1.5 hours, collect decoction liquor, then add 500mL water to decoct for the second time 40 minutes, collection decoction liquor, then add 500mL water to decoct for the third time 40 minutes, collection decoction liquor, when each decoction, the backflow distilling out is reclaimed in condensation, and above-mentioned three decoction liquor and backflow are merged, and 70 ℃ of water-baths are concentrated into 150mL and filter to obtain filtrate, it is 95% ethanol that filtrate adds concentration, make concentration of alcohol reach 60%, after fully stirring, obtain alcohol deposit fluid, alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 36 hours, centrifugal 5 minutes of 3000r/min, get 70 ℃ of water-baths of supernatant, volatilization goes to add concentration 95% ethanol after ethanol again, make concentration of alcohol reach 80%, again alcohol deposit fluid is placed in to 4 ℃ of refrigerators, cold preservation 36 hours, filter paper filters to obtain filtered solution, 70 ℃ of water-baths of filtered solution, ethanol is removed in volatilization, filtered solution adds distilled water to 100mL, making solution contain crude drug amount is 1g/mL, obtain filtrate with 0.25 micron of filtering with microporous membrane, filtrate steam sterilization 30min at 121 ℃ obtains Chinese medicine extraction liquid,
The Chinese medicine extraction liquid preparing and triethanolamine are mixed to get to water; After water is mixed with oil phase (stearic acid, glyceryl monostearate, vaseline or paraffin), add again transdermal agent (azone, 1,2-PD, dimethyl sulfoxide or acetylated monoglyceride), obtain Chinese medicine transdermal agent.
The isolation identification test of test example 1 bovine subclinical mastitis pathogen
1, test material and method
1.1 test reagents and medicine
The Fossmatic of FOX company of Denmark 5000 serial somatic cell Quick testing instruments, Wanda Mountain dairy industry group provides.
Nutrient agar, Mai Kangkai culture medium, triple sugariron culture medium, Yin Hong methylene blue culture medium, nutrient broth, MH meat soup etc., purchased from Ao Bai star bio tech ltd, Shanghai.
Micro biochemical pipe: the antibacterial micro biochemical reaction tubes such as hydrogen sulfide, mannitol, sucrose, 0.1% methylene blue Lac Bovis seu Bubali, sorbitol, gelatin, carbamide, arabinose, 6.5% high salt meat soup, Esculin, lactose, 40% bile broth, purchased from sky, Hangzhou and microorganism reagent company limited.Lot number: 20090615.
Antibiotics drug sensitive test paper: benzylpenicillin, amoxicillin, ceftazidime, cefazolin sodium, ceftriaxone, cefoperazone, erythromycin, chloromycetin, lincomycin, gentamycin, ciprofloxacin, enrofloxacin, ofloxacin, doxycycline etc., be all purchased from Beijing the Temple of Heaven Pharmaceutical Biotechnology development company.Lot number: 0909181.
1.2 test method
Get the milk sample of aseptic collection and inoculate nutrient agar, rabbit blood culture-medium and broth medium, cultivate 24h for 37 ℃, cultivate 1-3d to not growing the culture medium continuation slowly of bacterium or bacteria growing.The growth performances such as the shape of observed and recorded antibacterial, size, edge, surface texture, protuberance degree, color, transparency, hardness, haemolysis.Sterile working hooks up single bacterium colony, Gram’s staining, microscopy.Change and microscopy result according to clinical data, newborn sample, adopt different differential mediums and cultural method to differentiate.
1.2.1 the evaluation of pathogen
1.2.1.1 Gram’s staining
(1) smear is made
The pathogenic bacterium of suspecting in culture medium after cultivating are put at microscope slide middle section after connecing collarium picking, get normal saline one after another drop of on bacterium colony, be evenly coated with connecing collarium, smear is fixed through Alcohol Flame, then smear is dyeed.
(2) dying operation step
1. drip reagent 1(ammonium oxalate crystal violet dye liquor in the dry fixed area of smear), first stain dyeing 1mi n, then uses distilled water flushing.
2. drip reagent 2(iodine liquid dye liquor) after mordant effect 1min, then use distilled water flushing, suck moisture with filter paper.
3. drip reagent 3 depigmenting agents (95% ethanol depigmenting agent), now slide should be shaken frequently, until come off without purple, decolouring 30s, with distilled water cleaning, current can not be too large.
4. drip the luxuriant red colouring agent of reagent 4() to redye after dyeing 30s, distilled water cleans, and waits to do microscopy.
(3) result is judged:
Gram positive bacteria is purple, and gram negative bacteria is red.
Microscopy result preliminary judgement is divided into gram-positive cocci and bacillus, Gram-negative coccus and bacillus.
1.2.1.2 the biochemical identification of pathogen
The antibacterial of pure culture is carried out to the biochemical test inspections such as hemolytic test, various sugar fermentating test (glucose, lactose, maltose, mannitol culture medium), hydrogen peroxide test, triple sugar iron test, 6.5%NaCl test, indole test, hydrogen sulfide production test, Esculin test, hippuric acid test, M.R test, motoricity test.
1.2.1.3 the animal experiment of pathogen
Only get the liquid culture intraperitoneal inoculation white mice 0.5mL/ of pathogen.Every kind of antibacterial is established 3 and repeats white mice and a matched group (intraperitoneal inoculation 0.5mL 0.9%NaCl), the death condition of white mice in observation statistic record 24-48h after inoculation, and the internal organs of getting firm dead white mice carry out the test of antibacterial isolation identification.
1.2.2 antibiotics drug sensitive test
Measure the sensitivity of the Antibiotics medicine of the bacterial strain that isolation identification has gone out with agar disk diffusion method (K-B method).After getting antibacterial pure culture, be inoculated in LB fluid medium, 10h is cultivated in 37 ℃ of concussions, draw appropriate bacterium liquid (about 108CFU/mL) with the cotton swab after sterilizing, evenly coat MH agar plate surface, branch is evenly placed the various antibiotics drug sensitive test papers of choosing, after 24h, by the diameter of slide gauge vertical survey inhibition zone, record measurement result (according to Products Show standard determination).
2, result of the test
2.1 bovine subclinical mastitis Isolation and identification of pathogens the results are shown in Table 1.
The isolation identification result of table 1 bovine subclinical mastitis pathogen
2.2 antibiotic medicine sensitivity tests results
Staphylococcus aureus is high to cefoperazone, ceftriaxone, ofloxacin, ciprofloxacin, enrofloxacin sensitivity, and benzylpenicillin, lincomycin, amikacin are had to obvious resistance.
Staphylococcus saprophyticus is high to ceftriaxone, ofloxacin, ciprofloxacin, enrofloxacin sensitivity, and benzylpenicillin, streptomycin, amoxicillin are had to obvious resistance.
Staphylococcus epidermidis is high to ceftriaxone, enrofloxacin, ofloxacin sensitivity, and benzylpenicillin, streptomycin, gentamycin are had to obvious resistance.
Streptococcus dysgalactiae, to high to ceftriaxone, enrofloxacin, ofloxacin sensitivity, has obvious resistance to benzylpenicillin, gentamycin, streptomycin.
Streptococcus agalactiae is high to ceftriaxone, enrofloxacin, ciprofloxacin, ofloxacin sensitivity, and benzylpenicillin, streptomycin, doxycycline are had to obvious resistance.
Streptococcus uberis is high to ceftriaxone, enrofloxacin, ofloxacin, ciprofloxacin sensitivity, and penicillin, gentamycin, streptomycin, lincomycin, amoxicillin are had to obvious resistance.
Escherichia coli is high to ceftriaxone, cefoperazone, ceftazidime, enrofloxacin, ciprofloxacin, ofloxacin, gentamycin sensitivity, and benzylpenicillin, lincomycin, streptomycin are had to obvious resistance.
Corynebacterium is high to ceftriaxone, enrofloxacin, ofloxacin sensitivity, and benzylpenicillin, streptomycin, lincomycin, amoxicillin, doxycycline are had to obvious resistance.
Salmonella is high to ceftriaxone, cefoperazone, enrofloxacin, ofloxacin sensitivity, and penicillin, lincomycin, streptomycin amoxicillin are had to obvious resistance.
Bacillus cercus is high to ceftriaxone, ceftazidime, enrofloxacin, ofloxacin, ciprofloxacin sensitivity, and amikacin, benzylpenicillin, doxycycline, lincomycin are had to obvious resistance.
Test example 2 Chinese medicine composition vitro antibacterial activity test of the present invention
1, test material
1.1 supply reagent thing: the Chinese medicine extraction liquid that embodiment of the present invention 1-8 is prepared;
The preparation of 1.2 Bacteria liquid: the Main Pathogenic Bacteria staphylococcus aureus, escherichia coli, streptococcus dysgalactiae of choosing mammitis of cow successively picking microbionation, in the test tube of 5mL MH meat soup, cultivated 12h for 37 ℃.Get 100 these culture fluid of μ L and join in 2mL MH meat soup, cultivate 12h for 37 ℃, get bacterium liquid appropriate, by 3-5mL normal saline corrected concentrations to 0.5 Maxwell standard, for subsequent use.Bacterium liquid after correction should be inoculated in 15min.
2, bacteriostatic test method
Bacteriostatic test adopts in vitro 2 times of dilution methods of medicinal liquid and cultivates and carry out in conjunction with agar slant, get 10, test tube, number consecutively is also arranged, in every pipe, add sterilizing broth bouillon 2mL, then in the 1st pipe, adding concentration is that 1g/mL is for reagent thing 2mL, mix rear absorption 2mL and add in the 2nd pipe, proceed to successively the 9th pipe and dilute, in the 9th pipe, discard 2mL.In the 10th pipe, do not add for reagent thing as antibacterial matched group.By backward front in the above-mentioned test organisms liquid 0.1mL that in vitro adds, be placed in 37 ℃ of calorstats and cultivate 24h.Observe in vitro bacterial growth situation.
Result judgement: if 1-9 pipe contains the minimum inhibitory concentration (MIC) as this bacterium containing concentration of the highly diluted multiple of medicinal liquid after cultivating.Then above-mentioned 1-9 pipe culture is inoculated in and on agar slant, cultivates 24h again with inoculating loop, observe and have or not colony growth, be no more than the culture medium drug level of 5 as the minimum bactericidal concentration (MBC) of this prescription using clump count, test repeats 2 times.
3, result of the test
Confession reagent thing (Chinese medicine extraction liquid) In Vitro Bacteriostasis mainly measures escherichia coli MIC and MBC is 62.5mg/mL, and staphylococcus aureus MIC and MBC are 31.2mg/mL, and streptococcus dysgalactiae MIC and MBC are 31.2mg/mL.
The clinical observation on the therapeutic effect test of test example 3 traditional Chinese medicine composition for treating mammitis of cow of the present invention
Choosing of 1 ill milch cow
Select 3 medium-and-large-sized cattle farms in Harbin City, cattle farm breeding milk cow kind is holstein cow, in the 1-12 month in 2009, in the cattle farm of selecting, remove the sick cattle of clinic mastitis, monthly carry out somatic cell from milk counting to observing normal healthy cow lactation period.By somatic number 15 × 10
5-150 × 10
5individual/mL latent mammitis milch cow is classified treatment target as.Be divided at random three groups, every group of 12 cow heads: Chinese medicine transdermal agent treatment group (supplying reagent thing group), ofloxacin group, massage matched group merely.
Table 1 latent mammitis detection method and criterion
2, for reagent thing: the Chinese medicine transdermal agent that embodiment of the present invention 1-8 is prepared.
3, therapeutic scheme
Choose somatic cell from milk count detection 15 × 10
5individual/mL-150 × 10
5individual/mL suffers from cattle.For fear of the stress of administration, supply reagent thing (Chinese medicine transdermal agent of the present invention) treatment group after milk cow milking finishes, before medication, clean the corresponding ill newborn district of disease cattle with warm water, after drying with dry towel, smear for reagent thing (Chinese medicine compound transdermal agent) 20-30mL in each newborn district, smears on one side, general massage, 2 times/d, 15-20min/ time, 5d is 1 course for the treatment of; Ofloxacin group is after milking, and with ofloxacin injection 50mL intramuscular injection, every day 2 times, 5d is 1 course for the treatment of; The case being in a bad way can continuous use 2 courses for the treatment of, observe therapeutic effect.
Simple massage group is being milked after end at every turn, simple massage cream district 15-20min.
4, curative effect determinate standard
Treat after 1-2 the course for the treatment of, judge according to the change of milk production of cow and milk quality.
Cure: the normal and milk yield of milk perusal is recovered, spiritual, appetite is normal.Somatic cell from milk count detection, Methods of Milk Somatic Cell Images number is reduced to 5 × 10
5individual/below mL.
Effective: the normal and milk production of cow of milk perusal recovers on a small quantity, and god is smart, appetite is normal.Somatic cell from milk count detection, Methods of Milk Somatic Cell Images number decreases before test, but still higher than 5 × 10
5individual/mL.
5, clinical observation result
Chinese medicine transdermal agent of the present invention is used for clinical therapeutic effect in table 2.Chinese medicine transdermal agent treatment group is after 1-2 the course for the treatment of, and what cure completely is 12 cow head 13Ge Ru districts, and newborn district cure rate is 81.25%; Breast district significant figure is 14, and newborn district effective percentage is 87.50%, does not cure case clinical symptoms and obviously alleviates.At 1, what cure completely the course for the treatment of is 12 cow head 11Ge Ru districts to ofloxacin group case, and newborn district cure rate is 78.57%; Breast district significant figure is 12, and newborn district effective percentage is 85.71%.Simple massage group case cure rate is lower, and newborn district cure rate is 16.67%; Breast district significant figure is 5, and newborn district effective percentage is 41.67%.
The clinical observation on the therapeutic effect result of the test of table 2 traditional Chinese medicine composition for treating mammitis of cow of the present invention
As can be seen from Table 2, cure rate and the effective percentage difference not obvious (P > 0.05) of Chinese medicine transdermal agent group of the present invention and ofloxacin group, Chinese medicine transdermal agent group of the present invention and the cure rate and extremely significantly (the P < 0.01) of effective percentage difference that massage matched group.
In addition, the prepared Chinese medicine transdermal agent of embodiment of the present invention 6-8 will exceed the about 7-8 percentage point of Chinese medicine transdermal agent of embodiment 1-5 on the whole for the cure rate of cow recessive mastitis.
Clinical observation on the therapeutic effect result shows, Chinese medicine composition of the present invention is the Chinese prescription of effectively treating clinically latent mammitis, and clinical observation has no any toxic and side effects, clinical use safety.
6, apply test
Adopt Chinese medicine transdermal agent of the present invention to treat to suffering from the sick cattle of latent mammitis, cure over thousands of cow head, cure rate reaches more than 90%.
Claims (8)
1. a Chinese medicine composition for prevention or treatment mammitis of cow, is characterized in that, is made up: Flos Carthami 20-25 part of each crude drug of following weight portion, Radix Sanguisorbae 5-10 part, Galla Chinensis 5-10 part, Fructus Chebulae 5-10 part, Rhizoma Coptidis 15-20 part, Radix Bupleuri 5-10 part and Herba Violae 10-15 part.
2. according to Chinese medicine composition claimed in claim 1, it is characterized in that, the weight portion of each crude drug is: 22 parts, Flos Carthami, 8 parts of Radix Sanguisorbaes, 8 parts of Galla Chinensiss, 8 parts of Fructus Chebulaes, 18 parts of Rhizoma Coptidis, 8 parts of Radix Bupleuri, 12 parts of Herba Violaes.
3. according to the Chinese medicine composition described in claim 1 or 2, it is characterized in that: be prepared into any acceptable preparation clinically; Described preparation comprises external preparation, oral formulations or ejection preparation.
4. according to Chinese medicine composition claimed in claim 3, it is characterized in that: described external preparation comprises transdermal agent, unguentum or cataplasma; Described oral formulations is powder or decoction.
5. a method of preparing Chinese medicine composition described in claim 1 or 2, the method comprises the following steps:
(1) take each crude drug by described weight portion; (2) each crude drug is obtained to Chinese medicine extraction liquid with water extraction; (3) Chinese medicine extraction liquid and triethanolamine are mixed to get to water, by adding again transdermal enhancer after water and oil phase mixing, obtain transdermal agent.
6. in accordance with the method for claim 5, it is characterized in that, the preparation method of described Chinese medicine extraction liquid comprises: (a) each crude drug water extraction is obtained to extracting solution; By concentrated extracting solution, filtration; (b) filtrate adds ethanol, and alcohol deposit fluid is centrifugal, gets supernatant; Supernatant adds ethanol to carry out precipitate with ethanol after vaporing away ethanol again, and alcohol deposit fluid is filtered, and filtered solution vapors away ethanol and adds distilled water again, with filtering with microporous membrane, obtains Chinese medicine extraction liquid.
7. it is characterized in that in accordance with the method for claim 5: described transdermal enhancer is azone, 1,2-PD, dimethyl sulfoxide or acetylated monoglyceride; Described oil phase comprises stearic acid, glyceryl monostearate, vaseline or paraffin.
8. the purposes of the Chinese medicine composition described in claim 1 or 2 in preparation prevention or treatment mammitis of cow medicine.
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