CN105055699A - Medicine for treating mycoplasmal pneumonia of swine and preparing method, detection method and application thereof - Google Patents
Medicine for treating mycoplasmal pneumonia of swine and preparing method, detection method and application thereof Download PDFInfo
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- 238000001514 detection method Methods 0.000 title abstract description 5
- 208000008977 mycoplasmal pneumonia of swine Diseases 0.000 title abstract 2
- 229940079593 drug Drugs 0.000 title description 35
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- 238000002360 preparation method Methods 0.000 claims description 28
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- 206010035664 Pneumonia Diseases 0.000 claims description 17
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- 238000013461 design Methods 0.000 claims description 8
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- 241001362585 Asterolepis Species 0.000 abstract description 5
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- 241001518143 Polypodium <fern> Species 0.000 abstract 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 abstract 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 abstract 3
- 241001251200 Agelas Species 0.000 abstract 1
- 238000002347 injection Methods 0.000 abstract 1
- 239000007924 injection Substances 0.000 abstract 1
- 239000000243 solution Substances 0.000 abstract 1
- 208000011580 syndromic disease Diseases 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 17
- 229930194936 Tylosin Natural products 0.000 description 10
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- WBPYTXDJUQJLPQ-VMXQISHHSA-N tylosin Chemical compound O([C@@H]1[C@@H](C)O[C@H]([C@@H]([C@H]1N(C)C)O)O[C@@H]1[C@@H](C)[C@H](O)CC(=O)O[C@@H]([C@H](/C=C(\C)/C=C/C(=O)[C@H](C)C[C@@H]1CC=O)CO[C@H]1[C@@H]([C@H](OC)[C@H](O)[C@@H](C)O1)OC)CC)[C@H]1C[C@@](C)(O)[C@@H](O)[C@H](C)O1 WBPYTXDJUQJLPQ-VMXQISHHSA-N 0.000 description 4
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to the field of veterinary medicine, in particular to medicine for treating mycoplasmal pneumonia of swine and a preparing method, a detection method and application thereof. The medicine is prepared from, by weight, 1-2 parts of polypodium asterolepis bak. and 1-2 parts of lycoris squamigera maxim and prepared into powder or granules. The preparing method includes the steps that the dry polypodium asterolepis bak. and the dry lycoris squamigera maxim are added with water with the weight 11 times that of the polypodium asterolepis bak. and the lycoris squamigera maxim for the first time to be soaked for 1 h and decocted for 1 h, and are added with water with the weight 4 times that of the polypodium asterolepis bak. and the lycoris squamigera maxim for the second time to be soaked for 1 h, water decoction is combined and filtered, filter liquor is concentrated and dried, and the obtained product is pulverized into fine powder, screened, mixed evenly and prepared into the powder. According to the chromatographic conditions of the detection method for medicine quality, an Agela C18 chromatographic column is adopted, the mobile phase comprises acetonitrile and a 0.5% acetic acid solution by the ratio of 30:70, the detection wavelength is 206 nm, the column temperature is 22 DEG C, the flow velocity is 1.0 mL/min, and the injection volume is 10 microliters. The medicine can further be used for treating porcine productive and respiratory syndrome.
Description
Technical field
The present invention relates to field of veterinary, be specifically related to active medicine of a kind of deer Herba Alii fistulosi and Lepisorus thunbergianus (Kaulf.) Ching and preparation method thereof.
Background technology
Swine enzootic pneumonia or mycoplasma pneumoniae of swine, also known as mycoplasma hyopneumoniae, are a kind of chronic respiratory infectious disease of pig.The main clinic symptoms of primary disease is cough and asthma, and pathological change position is mainly positioned at thoracic cavity.Lungs are major organs of pathological changes.Acute case is based on pulmonary edema and emphysema; Subacute and chronic cases is shown in pulmonary's " Macrobrachium nipponensis " sample consolidation.The speed of growth of morbidity pig is slow, and efficiency of feed utilization is low, and the feeding for fattening phase extends.
China's pig industry is just from extensive, form of raising scattered to scale, intensive future development, and the popular and harm of swine enzootic pneumonia is also on the rise.This disease all can occur throughout the year, and the equal easy infection of the pig at different cultivars, age.Ill pig poor growth, efficiency of feed utilization declines, and brings very large difficulty, also result in very large economic loss simultaneously to pig farm management and Disease epizootic.
At present, this disease is considered to participate in mycoplasma hyopneumoniae to cause a disease relevant.Many application antibiotic are treated clinically, the generation but antibiotic can not prevent infections, and once discontinue medication, disease will soon recur, and easily develops immunity to drugs.
Medicine of the present invention has effect of heat-clearing and toxic substances removing, expelling phlegm for arresting cough, depressed lung-energy dispersing, tonneau throat, is used for the treatment of swine enzootic pneumonia Be very effective.
As follows at the Ji Yuan of medicine Raw medicine of the present invention:
Lepisorus thunbergianus (Kaulf.) Ching is Polypodiaceae Herba Lepisori Thunbergiani platymiscium Lepisorus macrosphaerus (Bak.) Ching var. Asterolepis (Bak.) Ching
lepisorusasterolepis (Bak.) Ching [PolypodiumasterolepisBak.]root or herb.
Deer Herba Alii fistulosi is Amaryllidaceae lycoris plants deer Herba Alii fistulosi
lycorissquamigeraMaximbulb.
Summary of the invention
The object of the invention is for the deficiencies in the prior art, the medicine of the treatment swine enzootic pneumonia that a kind of prescription is simple, therapeutic effect is good is provided.
Another object of the present invention is to provide the preparation method of this medicine.
Present invention also offers the quality determining method of this medicine.
Present invention also offers the pharmaceutical applications of this medicine.
The object of the invention is by realize with under type:
Medicine is made up of the raw material of following weight portion: Lepisorus thunbergianus (Kaulf.) Ching 1 ~ 2 weight portion, deer Herba Alii fistulosi 1 ~ 2 weight portion.
This medicine is preferably made up of the raw material of following weight portion: Lepisorus thunbergianus (Kaulf.) Ching 1 weight portion, deer Herba Alii fistulosi 2 weight portion.
This medicine is preferably made up of the raw material of following weight portion: Lepisorus thunbergianus (Kaulf.) Ching 2 weight portion, deer Herba Alii fistulosi 1 weight portion.
This medicine is preferably made up of the raw material of following weight portion: Lepisorus thunbergianus (Kaulf.) Ching 1 weight portion, deer Herba Alii fistulosi 1 weight portion.
Powder, granule can be prepared in described medicine.
Described medicine is adopted and is prepared with the following method: get Lepisorus thunbergianus (Kaulf.) Ching, deer Herba Alii fistulosi, and mixing adds the water soaking 0.5 ~ 2 hour of 3 ~ 15 times amount, decoct 0.5 ~ 2 hour, decoct 2 ~ 4 times, each 0.5 ~ 2 hour, extracting solution merges, and filters, and filtrate concentrates, dry, be ground into fine powder, add adjuvant, mixing, make powder, to obtain final product.
Described medicine is adopted and is prepared with the following method: get dry Lepisorus thunbergianus (Kaulf.) Ching, deer Herba Alii fistulosi, and first time adds 11 times amount water soaking 1 hour, decocts 1 hour, and second time adds 4 times amount soak by water 1 hour, merges decocting liquid, filter, filtrate concentrates, and dry, pulverize into fine powder, sieve, mixing, makes powder, to obtain final product.
Described medicine is adopted and is detected with the following method: adopt high performance liquid chromatography to carry out the assay of paeonol:
(1) chromatographic condition: adopt AgelaC
18chromatographic column; Mobile phase: ratio is acetonitrile-0.5% acetum of 20 ~ 40:60 ~ 80; Determined wavelength: 190 ~ 210nm; Column temperature: 15 ~ 25 DEG C; Flow velocity: 0.5 ~ 1.5mLmin
-1; Sample size: 5 ~ 20 μ L;
(2) reference substance solution preparation: it is appropriate that precision takes the paeonol reference substance being dried to constant weight, adds dissolve with methanol and make reference substance solution;
(3) preparation of need testing solution: precision takes medicine of the present invention, adds methanol, reflux, extracting solution reflux solvent is also concentrated into dry, and residue is dissolved in water, and extracts with water saturated n-butyl alcohol jolting, merge n-butanol extracting liquid, with ammonia solution washing, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol, shake up, filter, get filtrate, obtain need testing solution;
(4) measure: precision measures above-mentioned need testing solution, each 5 ~ 20 μ L of reference substance solution respectively, inject high performance liquid chromatograph, detect.
Described medicine is preferably adopted and is detected with the following method: adopt high performance liquid chromatography to carry out the assay of paeonol:
(1) chromatographic condition: adopt AgelaC
18chromatographic column; Mobile phase: ratio is acetonitrile-0.5% acetum of 30:70; Determined wavelength: 206nm; Column temperature: 22 DEG C; Flow velocity: 1.0mLmin
-1; Sample size: 10 μ L;
(2) reference substance solution preparation: it is appropriate that precision takes 80 DEG C of paeonol reference substances being dried to constant weight, adds dissolve with methanol and makes the reference substance solution of every 1mL containing 0.2mg;
(3) preparation of need testing solution: precision takes medicine 10g of the present invention, adds methanol 40mL, reflux 4h, extracting solution reflux solvent is also concentrated into dry, residue add water 10mL dissolve, extract 5 times with water saturated n-butyl alcohol jolting, each 20mL, merges n-butanol extracting liquid, washs 3 times with ammonia solution, each 15mL, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol and is transferred in 10mL volumetric flask, add methanol to scale, shake up, filter, get filtrate, obtain need testing solution;
(4) measure: precision measures above-mentioned need testing solution, each 10 μ L of reference substance solution respectively, inject high performance liquid chromatograph, detect.
Described medicine can be used for the medicine preparing treatment swine enzootic pneumonia, pig blue-ear disease.
experiment one: medicine of the present invention is to the observation of curative effect of swine enzootic pneumonia
1 materials and methods
1.1 medicine
1.1.1 Chinese crude drug is all purchased from Kang Ji chain pharmacy.
1.1.2 positive drug: tylosin phosphonate, Rui Pu (Tianjin) animal pharmaceutical estate company limited produces, and authentication code is veterinary drug word (2014) 020033034, purchased from prosperous veterinary drug shop, Lianyungang.
1.2 experimental animal
The sick pig that 20 build is identical, extremity perfect, body weight is about 70kg, main clinic symptoms for asthma, cough, dyspnea, in ventral breathing, body temperature without significant change, inappetence, listless.
1.3 the preparation of test medicine
Medicine of the present invention: prescription: Lepisorus thunbergianus (Kaulf.) Ching 50g, deer Herba Alii fistulosi 50g; Preparation method: get dry Lepisorus thunbergianus (Kaulf.) Ching 50g, deer Herba Alii fistulosi 50g and add 1100mL water soaking 1 hour for the first time, decoct 1 hour, second time adds 400mL, decocts 1 hour, merges decocting liquid, filters, and filtrate concentrates, and dry, pulverize, makes powder, obtain medicine of the present invention.
Drugs compared A: prescription: Lepisorus thunbergianus (Kaulf.) Ching 100g; Preparation method: get dry Lepisorus thunbergianus (Kaulf.) Ching 100g, first time adds 1100mL water soaking 1 hour, decocts 1 hour, and second time adds 400mL, decocts 1 hour, merges decocting liquid, filters, and filtrate concentrates, and dry, pulverize, makes powder, obtain drugs compared A.
Drugs compared B: prescription: deer Herba Alii fistulosi 100g; Preparation method: get dry deer Herba Alii fistulosi 100g, first time adds 1100mL water soaking 1 hour, decocts 1 hour, and second time adds 400mL, decocts 1 hour, merges decocting liquid, filters, and filtrate concentrates, and dry, pulverize, makes powder, obtain drugs compared B.
Making a definite diagnosis of 1.4 experimental animals
The sick sample of acute case is inoculated in solid medium, at the tiny bacterium colony of solid medium surface presentation distinctive " fried egg " shape, diameter is 0.1 ~ 1.0mm, smear, the female Sa of a Ji and Wright's staining, high power Microscopic observation is visible as ring-type, spherical, point-like multiform mycoplasma.
The grouping of 1.5 experimental animals and medication
The sick pig of making a definite diagnosis is divided into 4 groups, is respectively medicine group of the present invention, drugs compared A group, drugs compared B group, positive drug tylosin phosphonate group; Medicine group of the present invention, feeds by body weight, and 5g/kg medicine of the present invention, 1 time/d, is used in conjunction 7d; Drugs compared A group, feeds by body weight, 5g/kg drugs compared A, 1 time/d, is used in conjunction 7d; Drugs compared B group, feeds by body weight, 5g/kg drugs compared B, 1 time/d, is used in conjunction 7d; Positive drug tylosin phosphonate group to be fed 0.03g/kg tylosin phosphonate by body weight, 1 time/d, is used in conjunction 7d.Every morning, timing in afternoon are sterilized 1 time to pig house and surrounding, keep each group of pig feeding and management condition consistent.
1.6 therapeutic effect judge
(1) cure: after administration, the transference cures such as sick pig asthma, cough, dyspnea, spirit, appetite recover normal.
(2) effective: after administration, the symptoms such as sick pig asthma, cough, dyspnea are obviously alleviated, and spirit, appetite take a turn for the better.
(3) invalid: after administration, the symptoms such as sick pig asthma, cough, dyspnea and spirit, appetite are all unchanged, even increase the weight of to occur dead.
1.7 recurrence investigation
In order to determine the therapeutic effect of disease, after healing (discontinuing medication) 30d, the recurrence of the tested pig of follow-up investigation.If asthma, cough, dyspnea appear in the sick pig after curing, in ventral breathing, body temperature is without significant change, and inappetence, the symptom such as listless is then judged to be that swine enzootic pneumonia recurs.
2 results and analysis
2.1 treatment contrast tests
Result of the test is in table 1 ~ 2, and from table 1 ~ 2, drugs compared A group and drugs compared B group, within the 7th day, cure rate is 80%, and medicine group of the present invention and positive drug tylosin phosphonate group all reach 100%.Comprehensive analysis, 4 treatment groups all receive good therapeutic effect, and wherein tylosin phosphonate group is suitable with medicine group therapeutic effect of the present invention.
The statistical result of table 1 Cure
The effective situation statistical result of table 2 (effectively but do not cure)
2.2 recurrence status investigation
Result of the test is in table 3, and as shown in Table 3, drugs compared A group, drugs compared B group, tylosin phosphonate group relapse rate are 40%, and medication therapy groups of the present invention is substantially without recurrence.
Table 3 recurrence status survey result
3 discuss
In this test, in medicine of the present invention, all medicines share, effect of heat-clearing and toxic substances removing of building together, expelling phlegm for arresting cough, depressed lung-energy dispersing, tonneau throat.
Recurrence follow-up investigation result shows, drugs compared A group, drugs compared B group, tylosin phosphonate group relapse rate are apparently higher than medicine group of the present invention, its reason may be clinically to abuse for a long time tylosin phosphonate, pathogen is caused to create drug resistance in various degree, though make its clinical symptoms that can slow down asthma and avoid secondary infection, the infection of mycoplasma hyopneumoniae can not be stoped and once clinical manifestation of discontinuing medication will recover very soon.Adopt the method for antibiotic therapy of generally feeding can only prevent the outburst of asthma in pig farm and popular, fundamentally can not eliminate its harm.And life-time service antibiotic can cause meat to change, also can cause drug residue and not reach the requirement of nuisanceless meat.
4 conclusions
Swine enzootic pneumonia clinically in chronic process, be a kind of be difficult to cure and relapse rate very high disease, carry out larger economic loss to industrial belt of raising pigs.Adopt medicine of the present invention and tylosin phosphonate to carry out therapeutic effect contrast, result shows that medicine of the present invention is suitable with antibiotic to the therapeutic effect of swine enzootic pneumonia, and relapse rate is very low, pollutes little, noresidue, is the method for ideal treatment swine enzootic pneumonia.Along with people day by day increasing high-quality pork demand, production pollution-free food or Organic food become trend, and medicine of the present invention is treatment swine enzootic pneumonia is good choosing.
experiment two: the quality determining method research of medicine of the present invention
1 instrument and reagent
1.1 instrument
High performance liquid chromatograph (Agilent110 high performance liquid chromatograph and work station, G1311Aquat pump, G1314 UV-detector).
1.2 reagent
Paeonol (paeonol) reference substance (Chinese pharmaceutical biological product calibrating academy); Chinese medicine composition of the present invention; Chinese crude drug (Kang Ji chain pharmacy provides); Methanol (chromatograph alcohol, biochemical work auxiliary reagent factory, Shanghai); Other reagent are analytical pure.
2 methods and result
2.1 prescription
Lepisorus thunbergianus (Kaulf.) Ching 500g, deer Herba Alii fistulosi 500g.
2.2 preparation
Get dry Lepisorus thunbergianus (Kaulf.) Ching 500g, deer Herba Alii fistulosi 500g, first time adds 11000mL water soaking 1 hour, decocts 1 hour, and second time adds 4000mL, decocts 1 hour, merges decocting liquid, filters, and filtrate concentrates, and dry, pulverize, to obtain final product.
The assay of 2.3 paeonols (paeonol)
2.3.1HPLC chromatographic condition
Adopt AgelaC
18(4.0mm × 125mm, 5 μm) chromatographic column; Mobile phase: ratio is acetonitrile-0.5% acetum of 30:70; Determined wavelength: 206nm; Column temperature: 22 DEG C; Flow velocity: 1.0mLmin
-1; Sample size: 10 μ L; Under this chromatographic condition, reference substance and sample chromatogram peak are well, noiseless to mensuration without deer Herba Alii fistulosi negative control.
2.3.2 the preparation of reference substance solution
It is appropriate that precision takes 80 DEG C of paeonol reference substances being dried to constant weight, adds methanol and make the solution of every 1mL containing 0.2mg.
2.3.3 the preparation of need testing solution and negative controls
Precision takes medicine 10g of the present invention, adds methanol 40mL, reflux 4h, extracting solution reflux solvent is also concentrated into dry, residue add water 10mL dissolve, extract 5 times with water saturated n-butyl alcohol jolting, each 20mL, merges n-butanol extracting liquid, washs 3 times with ammonia solution, each 15mL, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol and is transferred in 10mL volumetric flask, add methanol to scale, shake up, filter, get filtrate and get final product; Separately do not contain the negative controls of deer Herba Alii fistulosi in the preparation of prescription ratio, be made in the same way of negative controls.
2.3.4 the drafting of standard curve
It is appropriate that precision takes 80 DEG C of paeonol reference substances being dried to constant weight, makes 10.4,20.8,41.6,83.2,166.4 μ gmL with methanol
-1the solution of series concentration, precision measures each 10 μ L of above-mentioned 5 kinds of strength solution respectively, injects high performance liquid chromatograph and measures.
Carry out linear regression with peak area ratio and concentration, obtaining regression equation is: A=21.2763C-0.1391, r=0.9999.Show that paeonol is at 10.4 ~ 166.4 μ gmL
-1in good linear relationship in concentration range.
2.3.5 stability test
Accurate absorption need testing solution 10 μ L, respectively at 0,1,2,4,8h sample introduction, and calculates paeonol content.In result 8h, RSD is 0.47%(n=5).Show that sample solution is stable in 8h.
2.3.6 replica test
By 5 parts, need testing solution preparation method parallel processing sample, measure paeonol content in accordance with the law and calculate.It is 0.11mgg that result records paeonol average content
-1, RSD is 1.1%.
2.3.7 Precision Experiment
Accurate absorption paeonol reference substance solution, repeats sample introduction 5 times, measures peak area in accordance with the law.Result RSD is 0.26%(n=5).Show that precision is better.
2.3.8 response rate experiment
Precision takes 6 parts, the sample of the same lot number of known paeonol content, adds appropriate paeonol reference substance solution, operate, measure in accordance with the law, calculate the response rate by under sample determination item by high, medium and low concentration is accurate respectively.Result average recovery rate is 100.8%, RSD is 0.48%(n=5).
2.3.9 sample size measures
Measure reference substance solution and need testing solution respectively appropriate, filter with microporous filter membrane, each sample introduction 10 μ L, measures 3 batch samples by above-mentioned chromatographic condition, parallel assay 5 times.By external standard method with the content of calculated by peak area need testing solution paeonol.This product should be containing paeonol and indicates 96% ~ 104% of content, in every 1g sample containing paeonol, must not be less than 0.13mg.3 batch sample content are respectively 100.1%(RSD=1.3%), 101.6%(RSD=1.7%), 100.8%(RSD=1.2%).
detailed description of the invention:
Embodiment 1, a kind of medicine for the treatment of swine enzootic pneumonia, this medicine is made up of the raw material of following weight portion: Lepisorus thunbergianus (Kaulf.) Ching 1 weight portion, deer Herba Alii fistulosi 2 weight portion.
Embodiment 2, a kind of medicine for the treatment of swine enzootic pneumonia, this medicine is made up of the raw material of following weight portion: Lepisorus thunbergianus (Kaulf.) Ching 1 weight portion, deer Herba Alii fistulosi 1 weight portion.
Embodiment 3, a kind of medicine for the treatment of swine enzootic pneumonia, this medicine is made up of the raw material of following weight portion: Lepisorus thunbergianus (Kaulf.) Ching 2 weight portion, deer Herba Alii fistulosi 1 weight portion.
Embodiment 4, as claimed in claim 1 medicine, this medicine is made up of the raw material of following weight portion: Lepisorus thunbergianus (Kaulf.) Ching 1.5 weight portion, deer Herba Alii fistulosi 1 weight portion.
Embodiment 5, the medicine in embodiment 1 ~ 4 described in any one, this medicine technology can be prepared into powder, granule routinely.
Embodiment 6, the medicine in embodiment 1 ~ 4 described in any one, this medicine is adopted and is prepared with the following method: get Lepisorus thunbergianus (Kaulf.) Ching, deer Herba Alii fistulosi, mixing, adds the water soaking 0.5 ~ 2 hour of 3 ~ 15 times amount, decocts 0.5 ~ 2 hour, decoct 2 ~ 4 times, each 0.5 ~ 2 hour, extracting solution merges, filter, filtrate concentrates, dry, be ground into fine powder, add adjuvant, mixing, make powder, to obtain final product.
Embodiment 7, the medicine in embodiment 1 ~ 4 described in any one, this medicine is adopted and is prepared with the following method: get dry Lepisorus thunbergianus (Kaulf.) Ching, deer Herba Alii fistulosi, first time adds 11 times amount water soaking 1 hour, decocts 1 hour, and second time adds 4 times amount soak by water 1 hour, merge decocting liquid, filter, filtrate concentrates, dry, be ground into fine powder, sieve, mixing, make powder, to obtain final product.
Embodiment 8, the medicine described in embodiment 6 or 7, this medicine is adopted and is detected with the following method: adopt high performance liquid chromatography to carry out the assay of paeonol:
(1) chromatographic condition: adopt AgelaC
18chromatographic column; Mobile phase: ratio is acetonitrile-0.5% acetum of 20 ~ 40:60 ~ 80; Determined wavelength: 190 ~ 210nm; Column temperature: 15 ~ 25 DEG C; Flow velocity: 0.5 ~ 1.5mLmin
-1; Sample size: 5 ~ 20 μ L;
(2) reference substance solution preparation: it is appropriate that precision takes the paeonol reference substance being dried to constant weight, adds dissolve with methanol and make reference substance solution;
(3) preparation of need testing solution: precision takes medicine of the present invention, adds methanol, reflux, extracting solution reflux solvent is also concentrated into dry, and residue is dissolved in water, and extracts with water saturated n-butyl alcohol jolting, merge n-butanol extracting liquid, with ammonia solution washing, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol, shake up, filter, get filtrate, obtain need testing solution;
(4) measure: precision measures above-mentioned need testing solution, each 5 ~ 20 μ L of reference substance solution respectively, inject high performance liquid chromatograph, detect.
Embodiment 9, the medicine described in embodiment 6 or 7, this medicine is adopted and is detected with the following method: adopt high performance liquid chromatography to carry out the assay of paeonol:
(1) chromatographic condition: adopt AgelaC
18chromatographic column; Mobile phase: ratio is acetonitrile-0.5% acetum of 30:70; Determined wavelength: 206nm; Column temperature: 22 DEG C; Flow velocity: 1.0mLmin
-1; Sample size: 10 μ L;
(2) reference substance solution preparation: it is appropriate that precision takes 80 DEG C of paeonol reference substances being dried to constant weight, adds dissolve with methanol and makes the reference substance solution of every 1mL containing 0.2mg;
(3) preparation of need testing solution: precision takes medicine 10g of the present invention, adds methanol 40mL, reflux 4h, extracting solution reflux solvent is also concentrated into dry, residue add water 10mL dissolve, extract 5 times with water saturated n-butyl alcohol jolting, each 20mL, merges n-butanol extracting liquid, washs 3 times with ammonia solution, each 15mL, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol and is transferred in 10mL volumetric flask, add methanol to scale, shake up, filter, get filtrate, obtain need testing solution;
(4) measure: precision measures above-mentioned need testing solution, each 10 μ L of reference substance solution respectively, inject high performance liquid chromatograph, detect.
Embodiment 10, a kind of medicine for the treatment of swine enzootic pneumonia:
Prescription: Lepisorus thunbergianus (Kaulf.) Ching 50g, deer Herba Alii fistulosi 50g
Preparation method: get dry Lepisorus thunbergianus (Kaulf.) Ching 50g, deer Herba Alii fistulosi 50g and add 1100mL water soaking 1 hour for the first time, decoct 1 hour, second time adds 400mL, decocts 1 hour, merges decocting liquid, filters, and filtrate concentrates, drying.
Embodiment 11: pharmaceutical hard capsule agent of the present invention:
Drug prescription: Lepisorus thunbergianus (Kaulf.) Ching 500g, deer Herba Alii fistulosi 500g
Preparation method: get dry Lepisorus thunbergianus (Kaulf.) Ching 500g, deer Herba Alii fistulosi 500g, first time adds 11000mL water soaking 1 hour, decocts 1 hour, and second time adds 4000mL, decoct 1 hour, merge decocting liquid, filter, filtrate concentrates, dry, be ground into fine powder, add adjuvant, mixing, makes powder, makes 1000g.
High performance liquid chromatography is adopted to carry out assay to paeonol (paeonol):
(1) chromatographic condition: adopt AgelaC
18chromatographic column; Mobile phase: ratio is acetonitrile-0.5% acetum of 30:70; Determined wavelength: 206nm; Column temperature: 22 DEG C; Flow velocity: 1.0mLmin
-1; Sample size: 10 μ L;
(2) reference substance solution preparation: it is appropriate that precision takes 80 DEG C of paeonol reference substances being dried to constant weight, adds methanol and makes the solution of every 1mL containing 0.2mg;
(3) preparation of need testing solution: precision takes medicine 10g of the present invention, adds methanol 40mL, reflux 4h, extracting solution reflux solvent is also concentrated into dry, residue add water 10mL dissolve, extract 5 times with water saturated n-butyl alcohol jolting, each 20mL, merges n-butanol extracting liquid, washs 3 times with ammonia solution, each 15mL, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol and is transferred in 10mL volumetric flask, add methanol to scale, shake up, filter, get filtrate, obtain need testing solution;
(4) measure: precision measures above-mentioned need testing solution, each 10 μ L of reference substance solution respectively, inject high performance liquid chromatograph, detect, testing result is the content of paeonol is 0.1260mg/g.
Claims (10)
1. treat a medicine for swine enzootic pneumonia, it is characterized in that, this medicine is made up of the raw material of following weight portion: Lepisorus thunbergianus (Kaulf.) Ching 1 ~ 2 weight portion, deer Herba Alii fistulosi 1 ~ 2 weight portion.
2. medicine as claimed in claim 1, it is characterized in that, this medicine is made up of the raw material of following weight portion: Lepisorus thunbergianus (Kaulf.) Ching 1 weight portion, deer Herba Alii fistulosi 2 weight portion.
3. medicine as claimed in claim 1, it is characterized in that, this medicine is made up of the raw material of following weight portion: Lepisorus thunbergianus (Kaulf.) Ching 2 weight portion, deer Herba Alii fistulosi 1 weight portion.
4. as claim 1 medicine, it is characterized in that, this medicine is made up of the raw material of following weight portion: Lepisorus thunbergianus (Kaulf.) Ching 1 weight portion, deer Herba Alii fistulosi 1 weight portion.
5. as the medicine in Claims 1 to 4 as described in any one, it is characterized in that, this medicine is prepared into powder, granule.
6. medicine as claimed in claim 5, is characterized in that, this medicine is adopted and prepared with the following method: get Lepisorus thunbergianus (Kaulf.) Ching, deer Herba Alii fistulosi, mixing, adds the water soaking 0.5 ~ 2 hour of 3 ~ 15 times amount, decocts 0.5 ~ 2 hour, decoct 2 ~ 4 times, each 0.5 ~ 2 hour, extracting solution merges, filter, filtrate concentrates, dry, be ground into fine powder, add adjuvant, mixing, make powder, to obtain final product.
7. medicine as claimed in claim 6, is characterized in that, this medicine is adopted and prepared with the following method: get dry Lepisorus thunbergianus (Kaulf.) Ching, deer Herba Alii fistulosi, first time adds 11 times amount water soaking 1 hour, decocts 1 hour, and second time adds 4 times amount soak by water 1 hour, merge decocting liquid, filter, filtrate concentrates, dry, be ground into fine powder, sieve, mixing, make powder, to obtain final product.
8. as the medicine in Claims 1 to 4 as described in any one, it is characterized in that, this medicine is adopted and is detected with the following method: adopt high performance liquid chromatography to carry out the assay of paeonol:
(1) chromatographic condition: adopt AgelaC
18chromatographic column; Mobile phase: ratio is acetonitrile-0.5% acetum of 20 ~ 40:60 ~ 80; Determined wavelength: 190 ~ 210nm; Column temperature: 15 ~ 25 DEG C; Flow velocity: 0.5 ~ 1.5mLmin
-1; Sample size: 5 ~ 20 μ L;
(2) reference substance solution preparation: it is appropriate that precision takes the paeonol reference substance being dried to constant weight, adds dissolve with methanol and make reference substance solution;
(3) preparation of need testing solution: precision takes medicine of the present invention, adds methanol, reflux, extracting solution reflux solvent is also concentrated into dry, and residue is dissolved in water, and extracts with water saturated n-butyl alcohol jolting, merge n-butanol extracting liquid, with ammonia solution washing, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol, shake up, filter, get filtrate, obtain need testing solution;
(4) measure: precision measures above-mentioned need testing solution, each 5 ~ 20 μ L of reference substance solution respectively, inject high performance liquid chromatograph, detect.
9. medicine as claimed in claim 8, is characterized in that, this medicine is adopted and detected with the following method: adopt high performance liquid chromatography to carry out the assay of paeonol:
(1) chromatographic condition: adopt AgelaC
18chromatographic column; Mobile phase: ratio is acetonitrile-0.5% acetum of 30:70; Determined wavelength: 206nm; Column temperature: 22 DEG C; Flow velocity: 1.0mLmin
-1; Sample size: 10 μ L;
(2) reference substance solution preparation: it is appropriate that precision takes 80 DEG C of paeonol reference substances being dried to constant weight, adds dissolve with methanol and makes the reference substance solution of every 1mL containing 0.2mg;
(3) preparation of need testing solution: precision takes medicine 10g of the present invention, adds methanol 40mL, reflux 4h, extracting solution reflux solvent is also concentrated into dry, residue add water 10mL dissolve, extract 5 times with water saturated n-butyl alcohol jolting, each 20mL, merges n-butanol extracting liquid, washs 3 times with ammonia solution, each 15mL, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol and is transferred in 10mL volumetric flask, add methanol to scale, shake up, filter, get filtrate, obtain need testing solution;
(4) measure: precision measures above-mentioned need testing solution, each 10 μ L of reference substance solution respectively, inject high performance liquid chromatograph, detect.
10. as the application of the medicine in Claims 1 to 4 as described in any one in the medicine of preparation treatment pig blue-ear disease.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102697959A (en) * | 2012-05-08 | 2012-10-03 | 广州市和生堂动物药业有限公司 | Veterinary drug of traditional Chinese veterinary drug for preventing and treating porcine respiratory disease syndrome |
| CN104174003A (en) * | 2014-09-10 | 2014-12-03 | 施瑞客（天津）生物技术有限公司 | Traditional Chinese medicine composition for preventing and treating swine enzootic pneumonia and preparation method thereof |
| CN104622932A (en) * | 2015-02-06 | 2015-05-20 | 郭敏 | Traditional Chinese medicine for treating myocardial ischemia and preparation method, detection method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102697959A (en) * | 2012-05-08 | 2012-10-03 | 广州市和生堂动物药业有限公司 | Veterinary drug of traditional Chinese veterinary drug for preventing and treating porcine respiratory disease syndrome |
| CN104174003A (en) * | 2014-09-10 | 2014-12-03 | 施瑞客（天津）生物技术有限公司 | Traditional Chinese medicine composition for preventing and treating swine enzootic pneumonia and preparation method thereof |
| CN104622932A (en) * | 2015-02-06 | 2015-05-20 | 郭敏 | Traditional Chinese medicine for treating myocardial ischemia and preparation method, detection method and application thereof |
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Application publication date: 20151118 |