CN104042680A - Dicranostigma leptopodum extract perfusate, and preparation method and application thereof - Google Patents
Dicranostigma leptopodum extract perfusate, and preparation method and application thereof Download PDFInfo
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- CN104042680A CN104042680A CN201410241597.9A CN201410241597A CN104042680A CN 104042680 A CN104042680 A CN 104042680A CN 201410241597 A CN201410241597 A CN 201410241597A CN 104042680 A CN104042680 A CN 104042680A
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Abstract
The invention provides a preparation method of a Dicranostigma leptopodum extract perfusate. The method comprises the following steps: adding water to a Dicranostigma leptopodum extract, decocting, mixing filtrates, and concentrating to obtain a concentrate; and extracting with ethanol, removing ethanol, adding benzyl alcohol and Tween-80 to the above obtained ethanol removed medicinal liquid, adding injection water to a full dose, and sterilizing to obtain the Dicranostigma leptopodum extract perfusate. The invention also provides an application of the perfusate in the preparation of medicines for treating cow mastitis. The Dicranostigma leptopodum extract perfusate adopting the Dicranostigma leptopodum extract as an effective component has very good heat clearing, detoxifying, bacterium resisting, inflammation eliminating and virus resisting effects. The perfusate is a pure traditional Chinese medicinal preparation, contains no chemical components, and is in favor of the safety of dairy products. Products produced through extraction and preparation integrated production have the advantages of stable properties and controllable quality, and are in favor of industrial production. The perfusate has the advantages of definite effect, no residual or toxic side effects, and realization of use in the lactation period as an effective medicine for treating clinic mastitis of cows, and becomes a good substitute of antibiotics.
Description
Technical field
The invention belongs to veterinary drug technical field, be specifically related to a kind of Herba dicranostigmae extract infusion liquid.
Background technology
Herba dicranostigmae is Papaveraceae Herba Dicranostigmatis Leptopodi platymiscium.In north and south, the Qinling Mountains, Gansu Province, current territory, Weihe is widely distributed.No matter all herbal medicine of Herba dicranostigmae, or compound compatibility, all there are the effects such as heat-clearing and toxic substances removing, reducing swelling and alleviating pain, parasite killing, in clinical use, find that Herba dicranostigmae reaches more than 90% lymphoid tuberculosis, pulmonary tuberculosis, bone tuberculosis and sinus tract ulcer total effective rate, has obvious therapeutic effect to diseases such as various boils, fistula, intractable stomatitis, wound, herpes zoster, colpitis mycotica, suppurative otitis media, sheep aphtha and scabies.
Milch cow clinical mastitis is cow mammary gland tissue inflammation, is mainly caused by multiple unspecific pathogenic microorganism, as antibacterial etc.Wherein, antibacterial is mainly Streptococcus, staphylococcus, Corynebacterium and Enterobacter etc.; Fungus has Candida, Piedraia and Eurotium etc.But the highest with the bacterial bearing rate of staphylococcus aureus in the dairy cow milk of trouble mastitis, be secondly escherichia coli and streptococcus; Mostly the clinical mastitis being caused by virus is due to secondary infection.In addition, environmental factors and feeding and management also have a significant impact the sickness rate of milch cow clinical mastitis, when milch cow living environment sanitary condition is poor or the improper damage latex dust of milking, all can cause breast resistance to decline, make breast easily suffer pathogenic microorganism invasion and attack and clinical mastitis occurs.Clinical mastitis Symptoms: milch cow suffers from mastitis, and it is thin that the lighter shows milk. breast amount reduces. contain floccule or grumeleuse, breast redness, appetite/body temperature changes little.It is obvious that severe one is suffered from district, pain. harden; Milk is light gray, and milk yield obviously declines; Fervescence, appetite declines.The enlargement of the scorching affected part area of malignant galactophore, as hard as flint, skin of breast is mulberry, and pain is very obvious; Can not squeeze milk, can only extrude a small amount of yellow fluid or blood and slime; Fervescence, breathing/pulse is accelerated; Loss of appetite or absolutely useless.If treatment can cause suppurating not in time. lose lactogenic function even dead.Its harm give milk for mastitis cause fall, milch cow is discarded, sick cattle is even dead, adds that medical care expenses pays wages, serious economic loss is brought at provisions family; The pathogen of clinical mastitis also can be detrimental to health by milk simultaneously.Treat by applying heavy dose of antibiotic and sulfa drugs China cattle farm for a long time, also brings many new problems in obtaining good therapeutic effect, such as increasing and drug residue problem of fastbacteria, has a strong impact on the use safety of milk product.
Summary of the invention
The invention provides a kind of Herba dicranostigmae extract infusion liquid, for this reason, the present invention also provides its preparation method and application.Herba dicranostigmae extract infusion liquid of the present invention has good therapeutic effect to bovine mastitis.
The technical solution used in the present invention is as follows: a kind of preparation method of Herba dicranostigmae extract infusion liquid: Herba dicranostigmae is added water and decocted, and merging filtrate is concentrated, obtains concentrated solution; Use afterwards ethanol alcohol extraction, go in medicinal liquid, to add benzyl alcohol, tween 80 after ethanol, add to the full amount of water for injection, sterilization, obtains Herba dicranostigmae extract infusion liquid of the present invention.
Preferably, described in add water that to decoct be the water that adds 2-6 times of quality, decoct 1-4 hour, decoct 3 times.
Further, described in add water that to decoct be the water that adds 2 times of quality, decoct 1 hour, decoct 3 times.
Preferably, described ethanol alcohol extraction is first to add the ethanol cold preservation of 1 times of concentrated solution quality precipitation 4 days, filters and abandons precipitation, adds successively the ethanol of 2 times, 3 times concentrated solution quality in filtrate again, and cold preservation precipitation 2 days, abandons precipitate.
Adopt this alcohol extracting method can effectively extract the effective ingredient in Herba dicranostigmae.
Further, after described ethanol alcohol extraction, regulate pH to 10 with sodium hydroxide.
Preferably, described adding in medicinal liquid after benzyl alcohol, tween 80 and water for injection, regulate pH value to 6-7.
Preferably, described sterilization is 100 DEG C of sterilizings 30 minutes.
Second object of the present invention is to provide the Herba dicranostigmae extract infusion liquid that application said method prepares.
The 3rd object of the present invention is to provide the application of above-mentioned Herba dicranostigmae extract infusion liquid in the medicine of preparation treatment bovine mastitis.
The present invention has following beneficial effect:
1, the present invention adopts Herba dicranostigmae extract as effective ingredient, has good heat-clearing and toxic substances removing, anti-inflammation and antiviral effect.Simultaneously owing to adopting pure Chinese medicinal preparation, not containing any chemical medicine composition, therefore be conducive to the safety of milk product.
2, by extract and the integrated production of preparation, institute the product property of producing stablize and quality controllable, be very beneficial for suitability for industrialized production.
3, as a kind of effectively treatment milch cow clinical mastitis medicine, its definite effect, noresidue and toxic and side effects, and can use in lactation period, will become the succedaneum that antibiotic is good.
Detailed description of the invention
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique in following embodiment, if no special instructions, is conventional method.Test material used in following embodiment, if no special instructions, is and purchases available from routine biochemistry reagent shop.If no special instructions, " % " is mass percent.
embodiment 1
The preparation method of Herba dicranostigmae extract infusion liquid of the present invention is as follows:
1 kilogram of Herba dicranostigmae adds water and decocts three times, adds water 6 kilograms at every turn, and the time of decocting is 4 hours.After decocting, filter, merging filtrate, be concentrated into 1000 milliliters, let cool; Add 1000 milliliters of ethanol 4-5 DEG C of cold preservation precipitations four days, filter and abandon precipitate, reclaim ethanol; Add successively 2000 milliliters, 3000 milliliters ethanol, all at 4-5 DEG C, cold preservation precipitates two days, filters and abandons precipitate, reclaim ethanol, hydro-oxidation sodium regulates pH to 10, and at 4-5 DEG C, cold preservation is filtered for 24 hours, filtrate is heated to without alcohol taste in water-bath, in medicinal liquid, add 15 milliliters of benzyl alcohol of pharmaceutical grade cosolvent and 15 milliliters of tween 80s, inject water to 1000 milliliters, regulate pH to neutral 6-7, filter, embedding, sterilizing in 100 DEG C, 30 minutes, obtains Herba dicranostigmae extract infusion liquid.This product is brown clear and bright liquid, Herba dicranostigmae extractive content 30%, and every 50 ml solns are equivalent to 0.5 gram of crude drug.
embodiment 2
The preparation method of Herba dicranostigmae extract infusion liquid of the present invention is as follows:
1 kilogram of Herba dicranostigmae adds water and decocts three times, adds water 4 kilograms at every turn, and the time of decocting is 2 hours.After decocting, filter, merging filtrate, be concentrated into 1000 milliliters, let cool; Add 1000 milliliters of ethanol 4-5 DEG C of cold preservation precipitations four days, filter and abandon precipitate, reclaim ethanol; Add successively 2000 milliliters, 3000 milliliters ethanol, all at 4-5 DEG C, cold preservation precipitates two days again, filters and abandons precipitate, reclaim ethanol, hydro-oxidation sodium regulates pH to 10, and at 4-5 DEG C, cold preservation is filtered for 24 hours, filtrate is heated to without alcohol taste in water-bath, in medicinal liquid, add 15 milliliters of benzyl alcohol, 15 milliliters of tween 80s, inject water to 1000 milliliters, regulate pH to 6-7, filter, embedding, sterilizing in 100 DEG C, 30 minutes, obtains Herba dicranostigmae extract infusion liquid.This product is brown clear and bright liquid, Herba dicranostigmae extractive content 30%, and every 50 ml solns are equivalent to 0.5 gram of crude drug.
embodiment 3
The preparation method of Herba dicranostigmae extract infusion liquid of the present invention is as follows:
1 kilogram of Herba dicranostigmae adds water and decocts three times, adds water 2 kilograms at every turn, and the time of decocting is 1 hour.After decocting, filter, merging filtrate, be concentrated into 1000 milliliters, let cool; Add 1000 milliliters of ethanol 4-5 DEG C of cold preservation precipitations four days, filter and abandon precipitate, reclaim ethanol; Add successively 2000 milliliters, 3000 milliliters ethanol, all at 4-5 DEG C, cold preservation precipitates two days again, filters and abandons precipitate, reclaim ethanol, hydro-oxidation sodium regulates pH to 10, and at 4-5 DEG C, cold preservation is filtered for 24 hours, filtrate is heated to without alcohol taste in water-bath, in medicinal liquid, add 15 milliliters of benzyl alcohol, 15 milliliters of tween 80s, inject water to 1000 milliliters, regulate pH to 6-7, filter, embedding, sterilizing in 100 DEG C, 30 minutes, obtains Herba dicranostigmae extract infusion liquid.This product is brown clear and bright liquid, Herba dicranostigmae extractive content 30%, and every 50 ml solns are equivalent to 0.5 gram of crude drug.
embodiment 4 applies the test of Herba dicranostigmae extract infusion liquid of the present invention
1, extracorporeal bacteria inhibitor test
1.1 strain staphylococcus aureuses
84184, colon bacillus C
8321, salmonella C
7926: all purchased from China Veterinary Drugs Supervisory Inst.; Klebsiella
219, bacillus pyocyaneus
1014, Bacillus proteus
1345: all purchased from Beijing biologics qualification institute.
1.2 culture medium staphylococcus aureuses are cultivated with beef-protein medium, all the other strains plain agar culture medium culturing, and pH is 7.2-7.4; Doubling dilution is all used broth bouillon, and pH is 7.0.
1.3 methods are tested Herba dicranostigmae extract infusion liquid of the present invention in embodiment 1-3 by cup-plate method, and adopt SHUANGHUANGLIAN ZHUSHEYE that expedition Pharmaceutical in Hebei produces in contrast, and the size of inhibition zone represents the power of bacteriostasis; Measure the minimum inhibitory concentration of Herba dicranostigmae extract infusion liquid with doubling dilution, the height of Mlc represents the weak strong of bacteriostasis.
1.4 result
Determine that through preliminary experiment the Herba dicranostigmae extract infusion liquid fungistatic effect of embodiment 3 is best.The Herba dicranostigmae extract infusion liquid that following test all obtains with embodiment 3 is tested.
Result of the test shows, Herba dicranostigmae extract infusion liquid of the present invention all has an inhibitory action to staphylococcus aureus, Bacillus proteus, colon bacillus, bacillus pyocyaneus, salmonella and klebsiella, the results are shown in Table 1.
The In Vitro Bacteriostasis effect comparison of table 1 Chinese medicinal perfusion liquid
。
Can find out from extracorporeal bacteria inhibitor test and the antibacterial activity in vivo result of the test of Herba dicranostigmae extract infusion liquid, the bacteriostasis of Herba dicranostigmae extract infusion liquid of the present invention is strong compared with SHUANGHUANGLIAN ZHUSHEYE, particularly to colon bacillus, therefore milch cow clinical mastitis is had to therapeutical effect.
1.5, bacteriostasis
1.5.1 bacteriostatic experiment: adopt plate dilution method, Herba dicranostigmae extract infusion liquid is carried out to doubling dilution with sterile distilled water, be diluted to 0.30mg/ml, 0.26 mg/ml, 0.22 mg/ml, 0.18 mg/ml, 0.14 mg/ml, 6 concentration of 0.10 mg/ml, separately establish blank (CK) and measure.Then with aseptic straw, diluent is quantitatively joined respectively in sterile petri dish, fully mix with the culture medium of quantitatively pouring into, leave standstill to solidify and make containing medicinal liquid flat board.By experiment required dosage, draw respectively quantitative tested bacterium liquid with pipet, be applied on flat board, put cultivation 1-2d in 37 DEG C of calorstats, observe colony growth situation, calculate clump count with the method for plate culture count, separately get a series of flat boards that do not add any medicinal liquid as blank (CK).There is no the minimum alkaloid concentration of colony growth completely as the minimal inhibitory concentration (MIC) of this medicine using naked-eye observation, repeat to test 3 times.Bacteriostatic experiment the results are shown in Table 2.
The antibacterial result of table 2 variable concentrations Herba dicranostigmae extract infusion liquid
。
As shown in Table 2, the MIC of bacillus coli is 0.18mg/ml; The MIC of staphylococcus aureus is 0.14mg/ml; The MIC of enterococcus faecalis is 0.30mg/ml.
2, acute toxicity test
2.1 test drugs: Herba dicranostigmae extract infusion liquid prepared by the embodiment of the present invention 3.
60 of the healthy mices of 2.2 experimental animals: body weight 18-21g, male and female half and half.
2.3 test methods: white mice is divided into 5 dosage groups and 1 normal healthy controls group, 10 every group at random.Herba dicranostigmae extract infusion liquid of the present invention is made into doses concentration, intraperitoneal injection by geometric progression.Give observation post administration 7d, statistics mortality rate, calculates LD
50.
2.4 result of the tests and conclusion: give observation post administration 7d, add up mortality rate, calculate the LD of Herba dicranostigmae extract infusion liquid of the present invention by improvement karber's method
50for 3234mg/kg, 95% confidence interval is 3188.55-3299.66mg/kg.So belong to substantially nontoxic.
3, apply the concrete condition of Herba dicranostigmae extract infusion liquid of the present invention treatment clinical type dairy cow mastitis as follows:
Herba dicranostigmae extract infusion liquid prepared by 3.1 test drugs: embodiment of the present invention 1-3.Contrast medicine: SHUANGHUANGLIAN ZHUSHEYE 10ml, prop up, Jiangxi Chuangdao Animal Health Product Co., Ltd. produces.
3.2 clinical type dairy cow mastitis therapeutic tests
3.2.1 experimental animal: cow reproduction field, Duan Jia beach, Lanzhou and cattle farm, Fu Long level ground 12 examples (18Ge Ru district), the ill cattle of clinical mastitis.Making a definite diagnosis of mastitis: 1. outward appearance has red, swollen, hot, pain, milk is thin or have a blood streak; 2. LMT field diagnostic is made a definite diagnosis: get the newborn sample in garget milk district, select suspicious bacterium colony again and continue purification cultivation 48h after 37 DEG C of cultivation 24h, long bacterium person can assert aseptic, makes Bacteria Identification if any bacterial growth.
3.2.2 reagent: mastitis diagnosticum (LMT) is produced by Lanzhou Livestock and Animal Drug Inst., Chinese Academy of Agricultural Science; 70g/L blood agar plate and the self-control of steamed beef soup culture medium.
3.2.3 method: the solution (Herba dicranostigmae extract quality degree is 30%) that Herba dicranostigmae extract infusion liquid is poured into into every 50ml under aseptic condition, ill newborn district milk is extracted, inject Herba dicranostigmae extract infusion liquid 50ml, every day 2 times, within 3 days, be a course for the treatment of, contrast medicine to specifications method uses.
3.2.4 cure criterion: 1. clinical symptom disappearance, milk is normal; 2. LMT diagnosis is negative.
3.2.5 therapeutic outcome: with 12 of this medicine treatments, by example 1,2,3, prepare and control drug, each 3.Wherein, the Herba dicranostigmae extract infusion liquid of embodiment 3 is all cured 1 course for the treatment of, cure rate 100%; The Herba dicranostigmae extract infusion liquid of embodiment 1 and 2 and contrast medicine are all cured 1 example, and 1 example takes a turn for the better.Before treatment, get ill milk cattle breast sample and identify through antibacterial culturing, result wherein antibacterial is streptococcus and staphylococcus aureus.
Proved by above-mentioned therapeutic outcome, Herba dicranostigmae extract infusion liquid of the present invention is higher to the cure rate of clinical type dairy cow mastitis, and there is the features such as detumescence is fast, delactation is rapid, toxicity is low, and wherein the effect of the Herba dicranostigmae extract infusion liquid of embodiment 3 is best.
Finally it should be noted that: the foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, although the present invention is had been described in detail with reference to previous embodiment, for a person skilled in the art, its technical scheme that still can record aforementioned each embodiment is modified, or part technical characterictic is wherein equal to replacement.Within the spirit and principles in the present invention all, any amendment of doing, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (9)
1. the preparation method of a Herba dicranostigmae extract infusion liquid: it is characterized in that: Herba dicranostigmae is added water and decocted, and merging filtrate is concentrated, obtains concentrated solution; Use afterwards ethanol alcohol extraction, go in medicinal liquid, to add benzyl alcohol, tween 80 after ethanol, add to the full amount of water for injection, sterilization, obtains Herba dicranostigmae extract infusion liquid of the present invention.
2. method according to claim 1, is characterized in that: described in add water that to decoct be the water that adds 2-6 times of quality, decoct 1-4 hour, decoct 3 times.
3. method according to claim 2, is characterized in that: described in add water that to decoct be the water that adds 2 times of quality, decoct 1 hour, decoct 3 times.
4. method according to claim 1, it is characterized in that: described ethanol alcohol extraction is first to add the ethanol cold preservation of 1 times of concentrated solution quality precipitation 4 days, filters and abandons precipitation, adds successively the ethanol of 2 times, 3 times concentrated solution quality in filtrate again, cold preservation precipitation 2 days, abandons precipitate.
5. according to the method described in claim 1 or 4, it is characterized in that: after described ethanol alcohol extraction, regulate pH to 10 with sodium hydroxide.
6. method according to claim 1, is characterized in that: described adding in medicinal liquid after benzyl alcohol, tween 80 and water for injection, regulate pH value to 6-7.
7. method according to claim 1, is characterized in that: described sterilization is 100 DEG C of sterilizings 30 minutes.
8. application rights requires Herba dicranostigmae extract infusion liquid prepared by 1-7.
9. the application of the arbitrary described Herba dicranostigmae extract infusion liquid of claim 1-7 in the medicine of preparation treatment bovine mastitis.
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Cited By (1)
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|---|---|---|---|---|
| CN104523863A (en) * | 2014-12-03 | 2015-04-22 | 中国农业科学院兰州畜牧与兽药研究所 | Traditional Chinese medicine perfusate and preparation method and application of traditional Chinese medicine perfusate |
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Cited By (2)
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| CN104523863A (en) * | 2014-12-03 | 2015-04-22 | 中国农业科学院兰州畜牧与兽药研究所 | Traditional Chinese medicine perfusate and preparation method and application of traditional Chinese medicine perfusate |
| CN104523863B (en) * | 2014-12-03 | 2019-03-01 | 中国农业科学院兰州畜牧与兽药研究所 | A kind of traditional Chinese medicine infusion solution and its preparation method and application |
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Application publication date: 20140917 |