CN106389562A - Pudilan Xiaoyan granules, preparation method thereof, and product quality control method - Google Patents
Pudilan Xiaoyan granules, preparation method thereof, and product quality control method Download PDFInfo
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- CN106389562A CN106389562A CN201611083496.9A CN201611083496A CN106389562A CN 106389562 A CN106389562 A CN 106389562A CN 201611083496 A CN201611083496 A CN 201611083496A CN 106389562 A CN106389562 A CN 106389562A
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- coarse powder
- filtration
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Classifications
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Abstract
The invention belongs to the field of veterinary drugs, and discloses Pudilan Xiaoyan granules, a preparation method thereof and a product quality control method. The Pudilan Xiaoyan granules are prepared from the following raw materials: Mongolian dandelion herb, baical skullcap root, coptis root, bunge corydalis herb, indigowoad root, weeping forsythia capsule, gypsum and licorice root. The preparation method comprises the following steps: taking the raw medicinal materials, and preparing the raw medicinal materials into coarse powder respectively; adding ethanol into the baical skullcap root coarse powder to perform extraction for a plurality of times, performing filtration, combining filtrates, recovering the ethanol, and performing concentration to obtain a thick paste; adding water and a compound enzyme into the indigowoad root coarse powder, stirring uniformly, performing warm immersion for a plurality of times, performing filtration, combining filtrates, and performing membrane separation on the filtrates to obtain an indigowoad root aqueous extract; adding water into other raw medicinal material coarse powder, performing continuous countercurrent extraction in a boiling state, performing filtration, performing membrane separation on the filtrate to obtain a clarified solution, adding the indigowoad root aqueous extract, and performing concentration to obtain a thick paste; and mixing the thick paste with the baical skullcap root thick paste uniformly, adding auxiliary materials, mixing the materials uniformly, preparing granules, and performing drying to obtain the Pudilan Xiaoyan granules. The formula provided by the invention is reasonable, and high in absorptivity, can be added by virtue of drinking water, is convenient to use, greatly reduces labor intensity, and is beneficial for intensive and large-scale raising.
Description
Technical field
The invention belongs to veterinary medicine technical field is and in particular to blue antiphlogistic granules in a kind of Pu ground and preparation method thereof and product
Method of quality control.
Background technology
Infectious bronchitis of chicken (InfectiousBronchitis, IB) is by coronaviridae
（Coronaviridae）Coronavirus genuses（Coronavirus）Infectious bronchitis virus
A kind of highly contagious disease that (InfectiousBronchitisVirus, IBV) causes, is the weight in intensive chicken farm
Want one of epidemic disease.IBV is strong to the resistance of environment, can be permanently present in the environment of surrounding, once infection, is difficult to eradicate.
It is characterized in that diseased chicken cough, sneeze and trachea are trembled sound；Can also watery nasal discharge in chickling, and laying hen is laid eggs and reduces and quality change
Bad.This disease has the infectiousness of height, because cause of disease has many serotypes, and so that immunity inoculation is complicated.Infected chicken growth retardation,
Feed consumption increases, lays eggs and the decline of egg matter, death rate increase, causes big economic loss to poultry husbandry.
IB all can occur throughout the year, higher in cold, hyperpyrexia season sickness rate.Hot, cold, crowded, improper ventilation
And the shortage of vitamin, mineral, trace element all can promote the generation of primary disease.IBV is mainly through the air spittle and respiratory tract
Pass to susceptible chicken, spread speed is fast, and often whole hen house is fallen ill simultaneously.Additionally, IBV also by cloaca and contaminated can raise
Material, drinking-water, apparatus etc. make susceptible chicken infection morbidity through digestive tract.Remain to toxin expelling after diseased chicken rehabilitation to be up to 140 days as long as.
Vaccine prevention is the main method reducing IB morbidity, but IB is not effectively controlled.Concrete reason has：1. domestic
Popular serotype infectious bronchitis virus are different from other countries；2. the infectious bronchitis of external serotype
Scorching virus constantly incoming China so that this virus serotype of China numerous it is difficult to preventing and treating；3. domestic chicken group have simultaneously different shaped and
The avian infectious bronchitis virus of variation are common to be existed and popular.
There is no the specific drug for the treatment of IB at present, the measure anti-processed currently IB taken mainly has following four aspects：1. strengthen
Feeding and management, the factors such as artificial abortion, logistics that reduce lead to external source of disease invasion, such as forbid introducing a fine variety from epidemic-stricken area or unsafe chicken house,
Hatching egg could be hatched for 2 times with formaldehyde fumigation；2. specification environment sterilization, is used alternatingly aldehydes, phenols, acids, bases, surface work
Property the disinfectant such as agent class, oxidants, quick eliminate harmful source of disease in breeding environment；3. strict implement vaccination program, protects
Shield chicken group is not encroached on by IBV；4. find the IBV state of an illness, take counter-measure in time；As breathing pattern gives resolving phlegm and relieving asthma medicine, kidney
Type gives diuretic or electrolyte supplement, reduces damage to kidney etc.；5. experience Chinese prescription reply.
Content of the invention
For overcoming the shortcomings of the prior art, the purpose of the present invention aims to provide a kind of blue antiphlogistic granules in Pu ground
And preparation method thereof and product quality control method.
For achieving the above object, the technical scheme that the present invention takes is as follows：
A kind of Pu ground indigo plant antiphlogistic granules, are made up of the crude drug of following weight percentage ratio：Herba Taraxaci 20-40%, Radix Scutellariae 15-30%,
Rhizoma Coptidis 10-20%, Herba Corydalis Bungeanae 10-20%, Radix Isatidis 10-20%, Fructus Forsythiae 5-10%, Gypsum Fibrosum 5-10%, Radix Glycyrrhizae 5-10%.
Prescription mechanism of the present invention：In side the effect of Herba Taraxaci heat-clearing and toxic substances removing, dispersing swelling and dissipating binds, inducing diuresis for treating stranguria syndrome, therefore it is monarch drug；Yellow
A kind of reed mentioned in ancient books, Rhizoma Coptidis, Radix Isatidis nature and flavor bitter cold, heat-clearing and toxic substances removing, removing heat from blood sore-throat relieving, in order to for ministerial drug, to strengthen monarch drug heat-clearing and toxic substances removing, sore-throat relieving disappears
Swollen work(；Fructus Forsythiae dispelling wind and heat pathogens, heat-clearing and toxic substances removing, scattered carbuncle detumescence；Gypsum Fibrosum compatibility therewith, by means of Gypsum Fibrosum clearing heat in QI system, hinders testis ventilating machine
Effect, can make interior-heat and the heat of half exterior and half interior thoroughly send out to the greatest extent and solve；Use Radix Glycyrrhizae invigorating the spleen and replenishing QI, grow and cough lung moistening, emergency is detoxified, mediation hundred
Medicine.All medicines share, plays heat-clearing and toxic substances removing, anti-inflammation detumescence altogether, thus reversing pathogenesis, blocks patient's condition, cuts off the change of disease to nutrient blood.This product
Product from the multi-faceted regulation such as antiviral, immunomodulating, antiinflammatory, antipyretic-antalgic, anti-stress and can be treated, so that heat is clear, heresy is scattered,
Swollen all diseases such as disappear are alleviated.
Carry out after prescription according to herbal medicine theory, prescription of the present invention is to treat outside infectious bronchitis of chicken, has
Higher heat-clearing and toxic substances removing, anti-inflammation detumescence ability, Clinical practice is more effective.Pu ground of the present invention indigo plant antiphlogistic granules are in that brown color is extremely brown
The granule of color；Mildly bitter flavor, is mainly used in treating and preventing infectious bronchitis of chicken, granule is easy to use, beneficial to absorption.With
Method consumption：Mixed drink, every 1L water adds the blue antiphlogistic granules 2-8g in Pu ground, is used in conjunction 3-5 days.
Preparation method, step is as follows：
Take each crude drug in proportion, be respectively prepared coarse powder；The 60-70% ethanol extraction that Radix Scutellariae coarse powder is added 6-8 times of weight for several times,
1-2h every time, filtration, merging filtrate, reclaim ethanol and be concentrated into the thick paste of 60 DEG C of relative densities 1.30-1.32, standby；By plate
Blue root coarse powder adds the water of 6-8 times of weight and accounts for the compound enzyme of its percentage by weight 2-4%, stirs evenly, is heated to 50-70 DEG C of warm macerating number
Secondary, each 1-2h, filtration, merging filtrate, it is Radix Isatidis infusion that filtrate obtains clear liquor after membrance separation, standby；Remaining other
Crude drug coarse powder adds water continuous adverse current type extraction 0.5-1.5h, filtration under fluidized state of 6-8 times of weight, and filtrate is through membrance separation
Obtain clear liquor afterwards, add Radix Isatidis infusion, be condensed into the thick paste of 60 DEG C of relative densities 1.30-1.32；Mix with Radix Scutellariae thick paste,
Add adjuvant, mix, make granule, be dried, obtain final product the blue antiphlogistic granules in Pu ground.
Described continuous adverse current type is extracted as medical material and is moved in opposite directions in leaching container with solvent, continuously and fully
Carry out contacting a kind of method extracted.Forward feeding, reversely enter solvent, material is inversely continuously dynamically to flow with the flowing of solvent
Dynamic, enable medical material and solvent to keep relative motion, so that feed concentration is spread and update continuous action, and then ensure that feed liquid leaches speed
Degree is fast.
Preferably, the percentage by weight of described compound enzyme consists of：Cellulase 20-40%, trypsin 20-40%, shallow lake
Powder enzyme 10-30%.
Preferably, membrance separation described at two is ultra-filtration and separation, and material is polypropylene or polysulfones, and molecular cut off is 60,000-
100000.
Preferably, the percentage by weight of described adjuvant consists of：Sucrose 60-70%, dextrin 30-40%.
Product quality control method：Using the Herba Taraxaci in TLC method blue antiphlogistic granules to gained Pu ground, Radix Scutellariae, Rhizoma Coptidis, hardship
Herba Violae, Radix Isatidis and Fructus Forsythiae are differentiated；Moisture, granularity, melting, loading amount and limit test of microbe all meet《Middle Chinese
People republic veterinary drug allusion quotation》Under two annex granule items of version in 2010；Measure content and the flavonoid glycoside of baicalin using HPLC method
Class compound total content.
Beneficial effect：In the inventive method, crude drug is broken into coarse powder, is conducive to the dissolution of effective ingredient, compare decoction pieces
Extract and shorten extraction time；Multiplex-enzyme extraction temperature is low, and extract yield is high；Continuous adverse current type extracts to be had many advantages, such as：Extract speed
Degree is fast, extracts active ingredients are abundant, extract recovery rate height, solvent consumption is few, liquor strength is high, decrease evaporation and concentration etc. follow-up
Medicinal material coarse powder translational speed scalable in process skill, cylinder, thus length, the medical material of extraction time can be adjusted according to medical material feature
Extracted under gentle dynamic environment, heating-up temperature is relatively low, effective ingredient destruction is less, makes impurity content in medicinal liquid few,
Belong to continuous way to produce, disposal ability is big.Generally, the blue antiphlogistic granules in Pu ground, meet the novel chiral synthon category of country's issue；Pass through
Technological transformation, eliminates unnecessary impurity, increased active constituent content, beneficial to the absorption of fowl, thus improving treatment curative effect.
Additionally, granule reasonable recipe of the present invention, absorbance are high, can be added by drinking-water, easy to use, added by drinking-water, greatly
Reduce labor intensity greatly, be conducive to intensive, large-scale cultivation.
Specific embodiment
Below in conjunction with specific embodiment, technical scheme is described in further detail, but the protection model of the present invention
Enclose and be not limited thereto.
Embodiment 1
A kind of Pu ground indigo plant antiphlogistic granules, it is made up of following materials by weight percentage medicine：Herba Taraxaci 30%, Radix Scutellariae 20%, Rhizoma Coptidis
10%th, Herba Corydalis Bungeanae 10%, Radix Isatidis 10%, Fructus Forsythiae 10%, Gypsum Fibrosum 5%, Radix Glycyrrhizae 5%.
Preparation method：Take each crude drug in proportion, be respectively prepared coarse powder；Radix Scutellariae coarse powder is added the second of the 60v% of 7 times of weight
Alcohol first extracts 2h, filtration, and medicinal residues add the ethanol extraction 1h of the 60v% of 7 times of weight again, and filtration merges and extracts filtrate twice, reclaims
Ethanol is simultaneously concentrated into the thick paste of 60 DEG C of relative densities 1.31, standby；Radix Isatidis coarse powder is added the water of 7 times of weight and accounts for its weight hundred
Divide the compound enzyme than 3%, stir evenly, be heated to 60 DEG C of warm macerating 2h, filtration, medicinal residues add the water of 7 times of weight again and account for its percentage by weight
3% compound enzyme, stirs evenly, and is heated to 60 DEG C of warm macerating 2h, filtration, merges warm macerating filtrate twice, and filtrate obtains clear liquor after membrance separation
I.e. Radix Isatidis infusion, standby；Remaining other crude drug coarse powder adds water continuous adverse current type extraction under fluidized state of 7 times of weight
1h, filtration, filtrate obtains clear liquor after membrance separation, adds Radix Isatidis infusion, is condensed into the thick paste of 60 DEG C of relative densities 1.31；
Mix with Radix Scutellariae thick paste, add adjuvant, mix, make granule, be dried, obtain final product the blue antiphlogistic granules in Pu ground；Wherein, described compound enzyme
Percentage by weight consist of：Cellulase 35%, trypsin 35%, amylase 30%, the enzyme activity of cellulase is 20000U/
G, tryptic enzyme activity are 4000U/g, diastatic enzyme activity is 30000U/g；The percentage by weight of described adjuvant consists of：
Sucrose 65%, dextrin 35%；Membrance separation described at two is ultra-filtration and separation, and material is polypropylene, and molecular cut off is 80,000.
Embodiment 2
A kind of Pu ground indigo plant antiphlogistic granules, it is made up of following materials by weight percentage medicine：Herba Taraxaci 40%, Radix Scutellariae 15%, Rhizoma Coptidis
10%th, Herba Corydalis Bungeanae 10%, Radix Isatidis 10%, Fructus Forsythiae 5%, Gypsum Fibrosum 5%, Radix Glycyrrhizae 5%.
Preparation method：Take each crude drug in proportion, be respectively prepared coarse powder；Radix Scutellariae coarse powder is added the second of the 65v% of 6 times of weight
Alcohol first extracts 2h, filtration, and medicinal residues add the ethanol extraction 1h of the 65v% of 6 times of weight again, and filtration merges and extracts filtrate twice, reclaims
Ethanol is simultaneously concentrated into the thick paste of 60 DEG C of relative densities 1.30, standby；Radix Isatidis coarse powder is added the water of 6 times of weight and accounts for its weight hundred
Divide the compound enzyme than 2%, stir evenly, be heated to 60 DEG C of warm macerating 2h, filtration, medicinal residues add the water of 6 times of weight again and account for its percentage by weight
2% compound enzyme, stirs evenly, and is heated to 60 DEG C of warm macerating 2h, filtration, merges warm macerating filtrate twice, and filtrate obtains clear liquor after membrance separation
I.e. Radix Isatidis infusion, standby；Remaining other crude drug coarse powder adds water continuous adverse current type extraction under fluidized state of 6 times of weight
1h, filtration, filtrate obtains clear liquor after membrance separation, adds Radix Isatidis infusion, is condensed into the thick paste of 60 DEG C of relative densities 1.30；
Mix with Radix Scutellariae thick paste, add adjuvant, mix, make granule, be dried, obtain final product the blue antiphlogistic granules in Pu ground；Wherein, described compound enzyme
Percentage by weight consist of：Cellulase 30%, trypsin 40%, amylase 30%, the enzyme activity of cellulase is 20000U/
G, tryptic enzyme activity are 4000U/g, diastatic enzyme activity is 30000U/g；The percentage by weight of described adjuvant consists of：
Sucrose 60%, DEXTRIN %；Membrance separation described at two is ultra-filtration and separation, and material is polypropylene, and molecular cut off is 60,000.
Embodiment 3
A kind of Pu ground indigo plant antiphlogistic granules, it is made up of following materials by weight percentage medicine：Herba Taraxaci 20%, Radix Scutellariae 20%, Rhizoma Coptidis
15%th, Herba Corydalis Bungeanae 10%, Radix Isatidis 10%, Fructus Forsythiae 10%, Gypsum Fibrosum 10%, Radix Glycyrrhizae 5%.
Preparation method：Take each crude drug in proportion, be respectively prepared coarse powder；Radix Scutellariae coarse powder is added the second of the 70v% of 8 times of weight
Alcohol first extracts 2h, filtration, and medicinal residues add the ethanol extraction 1h of the 70v% of 8 times of weight again, and filtration merges and extracts filtrate twice, reclaims
Ethanol is simultaneously concentrated into the thick paste of 60 DEG C of relative densities 1.32, standby；Radix Isatidis coarse powder is added the water of 8 times of weight and accounts for its weight hundred
Divide the compound enzyme than 4%, stir evenly, be heated to 60 DEG C of warm macerating 2h, filtration, medicinal residues add the water of 8 times of weight again and account for its percentage by weight
4% compound enzyme, stirs evenly, and is heated to 60 DEG C of warm macerating 2h, filtration, merges warm macerating filtrate twice, and filtrate obtains clear liquor after membrance separation
I.e. Radix Isatidis infusion, standby；Remaining other crude drug coarse powder adds water continuous adverse current type extraction under fluidized state of 8 times of weight
1h, filtration, filtrate obtains clear liquor after membrance separation, adds Radix Isatidis infusion, is condensed into the thick paste of 60 DEG C of relative densities 1.32；
Mix with Radix Scutellariae thick paste, add adjuvant, mix, make granule, be dried, obtain final product the blue antiphlogistic granules in Pu ground；Wherein, described compound enzyme
Percentage by weight consist of：Cellulase 40%, trypsin 40%, amylase 2 0%, the enzyme activity of cellulase is 20000U/
G, tryptic enzyme activity are 4000U/g, diastatic enzyme activity is 30000U/g；The percentage by weight of described adjuvant consists of：
Sucrose 70%, dextrin 30%；Membrance separation described at two is ultra-filtration and separation, and material is polypropylene, and molecular cut off is 100,000.
Reference examples 1
This reference examples is with the difference of embodiment 1：Only by Herba Taraxaci, Radix Scutellariae, Herba Corydalis Bungeanae, Radix Isatidis four Chinese medicine material system
Become, and the part by weight between this four Chinese medicine material is with embodiment 1, i.e. Herba Taraxaci: Radix Scutellariae: Herba Corydalis Bungeanae: Radix Isatidis=3: 2: 1: 1.
Preparation method：With embodiment 1.
Control of product quality example 1
Character this product is brown color to tan granule；Mildly bitter flavor.
Differentiate
（1）Indentification by TLC for Herba Taraxaci in side：
The preparation of need testing solution：Take this product 5.0g, finely ground, plus methanol 30ml, supersound process 30 minutes, filtration, filtrate is evaporated,
The residue 30ml that adds water makes dissolving, filtration, and filtrate is extracted 3 times with chloroform shaking, and each 15ml discards chloroform liquid, water
Layer is extracted 2 times with ethyl acetate shaking, each 20ml, and combined ethyl acetate liquid is evaporated, residue methanol 1ml makes dissolving, as
Need testing solution.
The preparation of reference substance solution：Take caffeic acid reference substance, plus methanol makes the solution that every 1ml contains 0.5mg, as comparison
Product solution.
The preparation of negative solution：According to prescription medical material ratio and preparation method, preparation lacks the negative sample of Herba Taraxaci medical material, with confession
Test sample solution preparation method operates, as negative control solution.
Fixing phase：Silica gel g thin-layer plate.
Developing solvent：Butyl acetate-formic acid-water（7: 2.5: 2.5, volume ratio）.
Point sample amount：5-10μl.
Development distance：8-10cm.
Inspect：Launch, take out, dry, put ultra-violet lamp（365nm）Under inspect.
Conclusion：In embodiment 1 ~ 3 test sample chromatograph, on position corresponding with reference substance chromatograph, aobvious same color glimmering
Light principal spot, scarce Herba Taraxaci negative sample chromatograph is noiseless on a corresponding position.
（2）Indentification by TLC for Radix Scutellariae in side：
The preparation of need testing solution：Take this product 5.0g, finely ground, plus methanol 40ml, supersound process 20 minutes, filtration, filtrate is evaporated,
The residue 10ml slight fever that adds water makes dissolving, lets cool, and adjusts pH value to 1～2 with dilute hydrochloric acid, plus ethyl acetate shaking extracts 2 times, every time
15ml, united extraction liquid, it is evaporated, residue adds methanol 2ml makes dissolving, as need testing solution.
The preparation of reference substance solution：Take baicalin reference substance, plus methanol makes the solution that every 1ml contains 0.5mg, as comparison
Product solution.
The preparation of negative solution：According to prescription medical material ratio and preparation method, preparation lacks the negative sample of radix scutellariae medicinal materials, with for examination
Product solution preparation method operates, as negative control solution.
Fixing phase：Silica gel g thin-layer plate
Developing solvent：Ethyl acetate-butanone-formic acid-water（5: 3: 1: 1, volume ratio）.
Point sample amount：5-10μl.
Development distance：8-10cm.
Inspect：Launch, take out, dry, spray, with 1% ferric chloride ethanol solution, is put and inspected under daylight.
Conclusion：In embodiment 1 ~ 3 test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color
Point, scarce Radix Scutellariae negative sample chromatograph is noiseless on a corresponding position.
（3）Indentification by TLC for Rhizoma Coptidis in side：
The preparation of need testing solution：Take this product 5.0g, plus kieselguhr 2 g, finely ground, plus methanol 50ml, it is heated to reflux 1 hour, filter
Cross, filtrate, filtrate is concentrated into 2ml, as need testing solution.
The preparation of reference substance solution：Take berberine hydrochloride reference substance, plus methanol makes the solution that every 1ml contains 1mg, as right
According to product solution.
The preparation of scarce Rhizoma Coptidis feminine gender solution：According to prescription medical material ratio and preparation method, preparation lacks the negative control sample of Fructus Forsythiae,
Take the negative sample 5.0g of scarce Rhizoma Coptidis, by need testing solution preparation method, be obtained and lack Fructus Forsythiae feminine gender solution.
The selection of unfolding condition：Draw each 5-10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with
Toluene-ethyl acetate-isopropanol-strong ammonia solution-methanol（6: 3: 1.5: 1.5: 0.5, volume ratio）For developing solvent, put ammonia steam and satisfy
In the expansion cylinder of sum, presaturation 30 minutes, launches, takes out, dry, put ultra-violet lamp（365nm）Under inspect.
Conclusion：In embodiment 1 ~ 3 test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color
Point, scarce Rhizoma Coptidis negative sample chromatograph is noiseless on a corresponding position.
（4）Indentification by TLC for Fructus Forsythiae in side：
The preparation of need testing solution：Take this product 5.0g, finely ground, add water 20ml, ultrasonic 20 minutes, filtration, filtrate, use ethyl acetate
Shaking is extracted 2 times, each 20ml, and combined ethyl acetate liquid is evaporated, residue adds methanol 5ml makes dissolving, is added in neutral alumina column
（100-200 mesh, 6g, internal diameter 1cm）On, with 70v% ethanol 50ml eluting, collect eluent, be evaporated, residue add methanol 1ml make molten
Solution, as need testing solution.
The preparation of reference substance solution：Take phillyrin reference substance, plus methanol makes the solution that every 1ml contains 1mg, as reference substance
Solution.
The preparation of scarce Fructus Forsythiae feminine gender solution：According to prescription medical material ratio and preparation method, preparation lacks the negative control sample of Fructus Forsythiae,
By need testing solution preparation method, it is obtained and lacks Fructus Forsythiae feminine gender solution.
The selection of unfolding condition：Draw each 5-10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with
Chloroform-methanol（5: 1, volume ratio）For developing solvent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, at 105 DEG C
It is heated to mottle colour developing clear.
Conclusion：In embodiment 1 ~ 3 test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color
Point, scarce Fructus Forsythiae negative sample chromatograph is noiseless on a corresponding position.
（5）Indentification by TLC for Herba Corydalis Bungeanae in side：
The preparation of need testing solution：Take this product 5.0g, enriching ammonia solution moistens, plus chloroform 30ml, stands overnight, filtration,
Filtrate is evaporated, and residue adds chloroform 1ml makes dissolving, as need testing solution.
The preparation of reference substance solution：Take corynoline reference substance, plus methanol makes the solution that every 1m1 contains 1mg, as reference substance
Solution.
The preparation of negative solution：According to prescription medical material ratio and preparation method, preparation lacks the negative sample of Herba Corydalis Bungeanae medical material, with confession
Test sample solution preparation method operates, as negative control solution.
Fixing phase：The silica gel g thin-layer plate of 0.4%NaOH solution preparation.
Developing solvent：Hexamethylene-chloroform-methanol（7: 2: 1, volume ratio）
Point sample amount：5-10μl.
Development distance：8-10cm.
Inspect：Launch, take out, dry, spray is inspected with dilute bismuth potassium iodide test solution, daylight.
Conclusion：In embodiment 1 ~ 3 test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color
Point, scarce Herba Corydalis Bungeanae negative sample chromatograph is noiseless on a corresponding position.
（6）Thin layer research for Radix Isatidis in side
The preparation of need testing solution：Take this product 5.0g, finely ground, plus ethanol 20ml, supersound process 20min, filtration, filtrate is evaporated,
Residue adds Diluted Alcohol 1ml dissolving, as need testing solution.
The preparation of reference substance solution：Take arginine reference substance, plus ethanol makes the solution that every 1ml contains 0.5mg, as comparison
Product solution.
The preparation of negative solution：According to prescription medical material ratio and preparation method, preparation lacks the negative sample of chromatogram of Radix Isatidis, with confession
Test sample solution preparation method operates, as negative control solution.
Fixing phase：Silica gel g thin-layer plate.
Developing solvent：N-butyl alcohol-glacial acetic acid-water（19: 5: 5, volume ratio）
Point sample amount：5-10μl.
Development distance：8-10cm.
Inspect：Launch, take out, hot blast drying, spray, with ninhydrin solution, is heated to spot development at 105 DEG C clear.
Result：In embodiment 1 ~ 3 test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color
Point, negative control sample is noiseless on a corresponding position.
Control of product quality example 2
Press《Chinese veterinary pharmacopoeia》2010 editions two annex granules require and quality standard draft, have carried out character, grain to product
Degree, moisture, melting, loading amount, microbial limit and loading quantity inspection.Result see table 1.
Control of product quality example 3-- measures the content of baicalin
Assay method is set to：
【Instrument and reagent】High performance liquid chromatograph：Agilent Technologies 1260, Agilent chromatographic data works
Stand, ultraviolet variable-wavelenght detector（Anjelen Sci. ＆ Tech. Inc）.
Reference substance：Baicalin, lot number：110715-201318, content is 93.3%, purchased from Chinese food drug assay research
Institute.
The blue antiphlogistic granules in Pu ground：Embodiment 1, embodiment 2, the product of embodiment 3.
Methanol, acetonitrile are chromatographically pure, and other reagent all adopt analysis pure.
【Chromatographic condition and system suitability】It is filler with octadecylsilane chemically bonded silica；With 0.6%（g/mL）
Phosphoric acid solution-methanol（53: 47, volume ratio）For mobile phase；Detection wavelength is 278nm.Number of theoretical plate should not based on baicalin peak
Less than 2000.
【The preparation of reference substance solution】Take baicalin reference substance（Lot number：110715-201318, content is 93.3%, is purchased from
National Institute for Food and Drugs Control）In right amount, accurately weighed, plus methanol make every 1ml contain 60 g solution, obtain final product.
【The preparation of need testing solution】This product is finely ground, takes about 3 g, accurately weighed, plus 70% ethanol 40 mL, is heated to reflux 3
H, lets cool, filtration, and filtrate puts in 100 mL measuring bottles, and with 70% ethanol gradation washing container and residue, washing liquid is filtered into same measuring bottle
In, plus 70% ethanol, to scale, shakes up.Precision measures 1 mL, puts in 10 mL measuring bottles, plus methanol, to scale, shakes up, and obtains final product.
【Algoscopy】Accurate absorption reference substance solution and each 10 l of need testing solution, inject chromatograph of liquid, survey respectively
Fixed, obtain final product.
Product content measurement result such as table 2.
Zoopery example
1 materials and methods
1.1 trial drug
Test the 1st group～the 3rd group of test：Blue three groups of the high, medium and low dosage of antiphlogistic granules in embodiment 1 Pu ground.
Testing the 4th group is drug control group 1：The blue antiphlogistic granules in reference examples 1 Pu ground.
Testing the 5th group is drug control group 2：MAXINGSHIGANGSAN, Sichuan Heng Tong animal pharmaceutical Co. Ltd, lot number：
20160305.
1.2 experimental animals and virus
1 age in days Guangxi richness Salted chicken, male, it is purchased from Liang Feng agriculture and animal husbandry Co., Ltd of Nanning City.Raise and be for experiment to 15 ages in days.
Immune programme for children is：Chicken Marek's disease vaccine intramuscular injection during 1 age in days, during 7 age in days, newcastle disease IV is vaccine collunarium eye dripping, 14
Infections chicken cloacal bursa Attenuate vaccine collunarium eye dripping during age in days, each vaccine according to respective product description immunoprophylaxis is respectively
Can.Feedstuff is commercially available chickling complete feed, without antiviral drugs.
Avian infectious bronchitis viruses（IBV）：M41 standard strain, purchased from China Veterinary Drugs Supervisory Inst., lot number：AV1511.
Its EID of experimental determination50For 10-6.68/0.2mL.
M41 standard strain normal saline dilution concentration is 100 × EID50, the virus liquid having diluted adds penicillin and chain
The dual anti-concentration to penicillin in virus liquid of mycin is 100U/mL, the concentration of streptomycin is 0.1mg/mL, 4 DEG C of refrigerator overnight, warp
Allantoic cavity is inoculated in 7 age in days SPF Embryo Gallus domesticus, each egg inoculation 0.2mL, is incubated in 37 DEG C of constant incubators, discards dead in 24h
Die embryo, incubate to 72h, reached for the 3rd generation with chick embryo allantoic liquid, with asepsis injector collection chick embryo allantoic liquid in clean sterilizing flat board
In, standby.
1.3 test method
1.3.1 experimental animal packet and process
Using randomly assigne packet, 15 ages in days are only divided into 7 groups for examination chicken 210, every group 30.Wherein the 1st～3 group be high, in,
Three groups of low dosage, the 4th group is drug control group 1, and the 5th group is drug control group 2, and the 6th group is infection matched group, and the 7th group is
Blank control group.
1st～6 group of every chicken is with 100 × EID50IBV the 3rd generation chick embryo allantoic liquid collunarium eye dripping is contaminated, and 0.2mL/ is only.7th
Organize as blank control group, be left intact.
Chicken appearance depressed, the loss of appetite of spirit to be tested after counteracting toxic substances, loose random, cold, flock together, mouth breathing, asthma,
Tracheal sound, flow mucus in oral cavity, cough, sneeze, after the infectious bronchitis of chicken clinical symptoms such as amount of drinking water increase, by table 3
Listed usage and dosage administration.After drug withdrawal, the clinical manifestation of animal is tracked observing, lasts till observing time the 7th after drug withdrawal
My god.In infection before and observation terminate after test chicken is weighed.
1.3.2 the observation of clinical efficacy
（1）Observing time：Medication first starts to observe, and continues to observe 7 days after stopping administration.
（2）Clinical symptoms and record：During test, observe and record the clinical manifestation of chicken daily, such as spirit, appetite, exhale
Inhale road symptom, whether oral cavity flows mucus, amount of drinking water etc..Dissection test chicken when off-test, observed and recorded lungs, trachea,
The pathological change of trachea, nasal sinuses and nasal cavity etc..
1.3.3 the standard of curative effect evaluation
Cure rate：During test, clinical symptom disappearance after medication, spiritual and situation of searching for food is recovered normally to be judged to cure.Every group
The percentage rate that healing chicken number accounts for every group of test chicken number is cure rate.
Effective percentage：During test, clinical symptom relief after medication, spiritual and condition improved of searching for food is judged to effectively.Every group
The percentage rate that effectively chicken number accounts for every group of test chicken number is effective percentage.
Virus isolated rate：Dissection test chicken when off-test, randomly draws lungs, trachea and the trachea lower end of part chicken
Furcation, by prior art isolated viral, calculates virus isolated rate.Sample number/the sample of virus isolated rate=be separated to virus is total
Number.
1.3.4 curative effect synthetic determination
According to the difference analysis of clinical efficacy, pathological anatomy change etc., Comprehensive Assessment is made to the curative effect of this medicine.
1.4 data analysiss and process
Data analysiss and process are carried out using SPSS17.0 statistical software.Enumeration data adopts X2Inspection；Measurement data adopts t to examine
Test or variance analyses, P ＜ 0.05 is significant difference；P ＜ 0.01 is that difference is extremely notable.
1.5 results and analysis
1.5.1 clinical symptoms
Test chicken 48 h after counteracting toxic substances fall ill successively, and the sickness rate of each counteracting toxic substances group is 100%, involves rapidly full group, but all no chickens
Only dead, start after 72 h that obvious clinical symptoms occur, diseased chicken is mainly shown as lethargy, uneasy, happiness, loss of appetite,
Drink is intended to increase, and flocks together, gets rid of head, grab nose, sheds tears, watery nasal discharge, part chicken nasal sinuses swelling, and phenomenon of having loose bowels in indivedual chickens.Night can listen
To obvious cough and respiratory murmur.
Compared with infection matched group after 3 days by giving the blue antiphlogistic granules in the embodiment of the present invention 1 Pu ground of various dose, disease
Feelings mitigate, and spirit and appetite take a turn for the better, and night observation cough and sound reduce, wherein with the blue antiphlogistic granules high dose group in Pu ground
Good.Medication terminates latter first day, and the mental status observing each medication group test chicken is clearly better, and clinical symptoms substantially mitigate, night
Between observe and only have fragmentary cough, sound disappears.Observe within the 6th day after administration, the mental status of each medication group test chicken is basic
Recover normal, what different frequency in only fraction of chicken gets rid of a phenomenon.
The 7th day after administration, randomly draw 15 test chickens, jugular vein sacrificed by exsanguination for every group, anatomic observation each group is tested
The pathological changes of chicken organ organ.Result：There is substantial amounts of mucus, some mucus are in gel-shaped in infection matched group chicken oral cavity and nasal sinuses,
Trachea has obvious bleeding, congested phenomenon；There are obvious hyperemia, bleeding in pulmonary.The dissection disease of each treatment group test chicken
Become light compared with positive controls.
1.5.2 therapeutic effect
Efficacy result after being administered 5 days is shown in Table 4：The effect of the blue antiphlogistic granules in the embodiment of the present invention 1 each dosage Pu ground is substantially excellent
In the blue antiphlogistic granules in Pu ground of reference examples 1, the wherein curative effect of the blue antiphlogistic granules in the embodiment of the present invention 1 low dosage Pu ground and numb Fructus Pruni stone
Sweet scattered substantially suitable, but the curative effect of the blue antiphlogistic granules in the Pu of the embodiment of the present invention 1 middle and high dosage ground is substantially better than Maxingshigan
Dissipate.Result shows, Pu ground of the present invention indigo plant antiphlogistic granules to artificial challenge's infectious bronchitis of chicken disease therapeutic effect substantially, can use
Treatment in infectious bronchitis.
Claims (6)
1. a kind of Pu ground indigo plant antiphlogistic granules are it is characterised in that be made up of the crude drug of following weight percentage ratio：Herba Taraxaci 20-40%,
Radix Scutellariae 15-30%, Rhizoma Coptidis 10-20%, Herba Corydalis Bungeanae 10-20%, Radix Isatidis 10-20%, Fructus Forsythiae 5-10%, Gypsum Fibrosum 5-10%, Radix Glycyrrhizae 5-
10%.
2. a kind of method preparing the blue antiphlogistic granules in Pu as claimed in claim 1 ground it is characterised in that：Take each raw material in proportion
Medicine, is respectively prepared coarse powder；The 60-70% ethanol extraction that Radix Scutellariae coarse powder is added 6-8 times of weight for several times, each 1-2h, filtration, merge
Filtrate, reclaims ethanol and is concentrated into the thick paste of 60 DEG C of relative densities 1.30-1.32, standby；Radix Isatidis coarse powder is added 6-8 times of weight
Water and account for the compound enzyme of its percentage by weight 2-4%, stir evenly, be heated to 50-70 DEG C of warm macerating for several times, each 1-2h, filtration, close
And filtrate, it is Radix Isatidis infusion that filtrate obtains clear liquor after membrance separation, standby；Remaining other crude drug coarse powder adds 6-8 weight
Water again continuous adverse current type under fluidized state extracts 0.5-1.5h, filtration, and filtrate obtains clear liquor after membrance separation, adds Baphicacanthus cusia
Root infusion, is condensed into the thick paste of 60 DEG C of relative densities 1.30-1.32；Mix with Radix Scutellariae thick paste, add 2-4 times of thick paste gross weight
Adjuvant, mix, make granule, be dried, obtain final product the blue antiphlogistic granules in Pu ground.
3. preparation method as claimed in claim 2 is it is characterised in that the percentage by weight of described compound enzyme consists of：Fiber
Plain enzyme 20-40%, trypsin 20-40%, amylase 10-30%.
4. preparation method as claimed in claim 2 it is characterised in that：Membrance separation described at two is ultra-filtration and separation, and material is
Polypropylene or polysulfones, molecular cut off is 60,000-10 ten thousand.
5. preparation method as claimed in claim 2 is it is characterised in that the percentage by weight of described adjuvant consists of：Sucrose 60-
70%th, dextrin 30-40%.
6. a kind of blue antiphlogistic granules in Pu as claimed in claim 1 ground product quality control method it is characterised in that：Using TLC
Herba Taraxaci in method blue antiphlogistic granules to gained Pu ground, Radix Scutellariae, Rhizoma Coptidis, Herba Corydalis Bungeanae, Radix Isatidis and Fructus Forsythiae differentiate；Moisture,
Granularity, melting, loading amount and limit test of microbe all meet《Republic of China Veterinary Pharmacopoeia》Two annex of version in 2015
Under granule item；Measure the content of baicalin using HPLC method.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110057950A (en) * | 2019-04-12 | 2019-07-26 | 舟山市食品药品检验检测研究院 | A kind of construction method of the Pudilan sulfathiazole HPLC finger-print based on Detection wavelength switching |
| CN113125612A (en) * | 2021-05-15 | 2021-07-16 | 山东宏济堂制药集团股份有限公司 | Construction method of high performance liquid phase characteristic spectrum of corydalis bungeana |
| CN116898921A (en) * | 2023-08-10 | 2023-10-20 | 江西新世纪民星动物保健品有限公司 | Preparation method of cattail and dyer woad compound traditional Chinese medicine preparation |
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2016
- 2016-11-30 CN CN201611083496.9A patent/CN106389562A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110057950A (en) * | 2019-04-12 | 2019-07-26 | 舟山市食品药品检验检测研究院 | A kind of construction method of the Pudilan sulfathiazole HPLC finger-print based on Detection wavelength switching |
| CN110057950B (en) * | 2019-04-12 | 2021-06-22 | 舟山市食品药品检验检测研究院 | Detection wavelength switching-based Pudilan antiphlogistic tablet HPLC fingerprint construction method |
| CN113125612A (en) * | 2021-05-15 | 2021-07-16 | 山东宏济堂制药集团股份有限公司 | Construction method of high performance liquid phase characteristic spectrum of corydalis bungeana |
| CN116898921A (en) * | 2023-08-10 | 2023-10-20 | 江西新世纪民星动物保健品有限公司 | Preparation method of cattail and dyer woad compound traditional Chinese medicine preparation |
| CN116898921B (en) * | 2023-08-10 | 2024-05-31 | 江西新世纪民星动物保健品有限公司 | Preparation method of cattail and dyer woad compound traditional Chinese medicine preparation |
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