CN104983777A - Preparation method for radix astragali and licorice powder and product thereof - Google Patents
Preparation method for radix astragali and licorice powder and product thereof Download PDFInfo
- Publication number
- CN104983777A CN104983777A CN201510256517.1A CN201510256517A CN104983777A CN 104983777 A CN104983777 A CN 104983777A CN 201510256517 A CN201510256517 A CN 201510256517A CN 104983777 A CN104983777 A CN 104983777A
- Authority
- CN
- China
- Prior art keywords
- preparation
- radix astragali
- radix glycyrrhizae
- extract
- concentrated
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention relates to a preparation method for radix astragali and licorice powder and a product thereof. The method comprises the following steps that 1, radix astragali is used for preparing a radix astragali extract; 2, licorice roots are used for preparing a licorice root extract; and 3, the radix astragali extract and the licorice root extract are made into the radix astragali and licorice powder. According to the preparation method, the requirement for equipment is not high, the preparation technology is simple, and the production efficiency can be improved.
Description
Technical field
The present invention relates to preparation method of a kind of Radix Astragali Radix Glycyrrhizae powder and products thereof.
Background technology
The aviculture high speed development of current China, state of fowl sparetime university of world ranks are stepped into, but the occurrence and harm of China's fowl diseases is very serious, avian viral immunosuppressant disease is a class Important Infectious Diseases of puzzle aviculture, due to the existence of immunosuppressive condition, usually cause immune effect of vaccine difference or immuning failure, very easily concurrent and secondary animal infectious diease and parasitic disease, add sickness rate and the treatment difficulty of poultry, cause huge economic loss to aviculture.The immunostimulant kind of current use is a lot, comprises microorganism immunostimulant, as interferon; Chemical immunostimulant, as levamisole etc., there is the shortcomings such as high dose produces immunosuppressant, the residence time is long in body, Chinese traditional immunopoteniators has the advantages such as aboundresources, safe ready, complete function, noresidue and toxic and side effects are little, therefore, the active strong new medicine immunostimulant of Study and Development become in the urgent need to.
The active component of Radix Astragali Radix Glycyrrhizae powder is mainly polysaccharide and glycyrrhizic acid, and modern pharmacology research shows that astragalus polysaccharides and Angelica Polysaccharide have effect of enhancing human body immunity power, and glycyrrhizic acid has nonspecific immunity regulating action.Adopt traditional decoction and alcohol sedimentation technique to prepare Radix Astragali Radix Glycyrrhizae powder, also by most of polysaccharide active component removing while precipitate with ethanol remove impurity, cause Quality Down, affect clinical efficacy, and use a large amount of ethanol in precipitate with ethanol process, process costs is higher, and the time is longer; And when adopting water precipitating method as purification process, the impurity of removing is less, the powder less stable of preparation.
For effectively removing impurity, increasing stability, and retaining polysaccharide active component, being necessary the preparation method setting up a kind of rational Radix Astragali Radix Glycyrrhizae powder.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of Radix Astragali Radix Glycyrrhizae powder, to reach the high and stay-in-grade effect of the active constituent contents such as polysaccharide.
The invention provides a kind of preparation method of Radix Astragali Radix Glycyrrhizae powder, comprise the following steps:
S1, prepares Radix Astragali extract with Milkvetch Root;
S2, prepares Radix Glycyrrhizae extract with licorice medicinal materials;
S3, is mixed with described Radix Astragali extract and described Radix Glycyrrhizae extract as Radix Astragali Radix Glycyrrhizae powder.
In a detailed description of the invention, the preparation process of described Radix Astragali extract comprises:
A) soaked by Milkvetch Root, concentrated after extracting, the proportion being preferably concentrated into concentrated solution is 1.10-1.15;
B) concentrated solution step a) obtained first processes with pectase and protease, then processes with chitosan-acetic acid solution;
C) by step b) mixture that obtains filters, and with alkali liquor, filtrate is adjusted to weak acid to neutral, being preferably adjusted to pH value is 5.0-7.0, then concentrates, is drying to obtain Radix Astragali extract.
In a detailed description of the invention, step a) in, described in be extracted as low-temperature reduced-pressure reflux, extract.Preferably, the condition of described low-temperature reduced-pressure reflux, extract, is: vacuum is-0.03-0.05MPa, and temperature is 60-70 DEG C, and extraction time is 2-4 time, and extraction time is 1-3h.In one embodiment, the condition of described extraction is: vacuum is-0.05MPa, at 70 DEG C, extracts 3 times, each 2h.Preferably, step a) in, by weight, the consumption of water be the 6-10 of Milkvetch Root doubly.
In a detailed description of the invention, in step b) in, the consumption of pectase is 1-2g/100mL concentrated solution.
In a detailed description of the invention, in step b) in, the consumption of protease is 1-2g/100mL concentrated solution.
In a detailed description of the invention, in step b) in, the mass ratio of pectase and protease is 1:2-2:1.
In a detailed description of the invention, in step b) in, the addition of chitosan-acetic acid solution and the volume ratio of concentrated solution are 0.15:1-0.2:1.
According to a detailed description of the invention, the preparation process of described Radix Glycyrrhizae extract comprises:
A) soak licorice medicinal materials with ammonia spirit, concentrated after extracting, the proportion being preferably concentrated into concentrated solution is 1.10-1.15;
B) by steps A) the concentrated solution adjust ph that obtains is 5.0-6.0, then first processes with pectase and protease, then processes with chitosan-acetic acid solution;
C) by step B) mixture of gained filters, and adjust ph is 5.0-7.0, concentrated, is drying to obtain Radix Glycyrrhizae extract.
According to a detailed description of the invention, described in be extracted as low-temperature reduced-pressure reflux, extract.Preferably, the condition of described low-temperature reduced-pressure reflux, extract, is: vacuum is-0.03-0.05MPa, and temperature is 60-70 DEG C, and extraction time is 2-4 time, and extraction time is 1-3h.In one embodiment, the condition of described extraction is: vacuum is-0.05MPa, at 70 DEG C, extracts 3 times, each 2h.
In a detailed description of the invention, steps A) in, in described ammonia spirit, the mass content of ammonia is 0.075% ~ 0.168%.
Preferably, steps A) in, by weight, the consumption of water is 6-10 times of Radix Glycyrrhizae Milkvetch Root.
In a detailed description of the invention, in step B) in, the consumption of pectase is 1-2g/100mL concentrated solution.
In a detailed description of the invention, in step B) in, the consumption of protease is 1-2g/100mL concentrated solution.
In a detailed description of the invention, in step B) in, the mass ratio of pectase and protease is 1:2-2:1.
In a detailed description of the invention, in step B) in, the addition of chitosan-acetic acid solution and the volume ratio of concentrated solution are 0.15:1-0.2:1.
In a detailed description of the invention, the mass ratio of described Milkvetch Root and licorice medicinal materials is 1:1.
According to a specific embodiment of the present invention, the preparation method of described Radix Astragali Radix Glycyrrhizae powder comprises:
1) soaked by licorice medicinal materials ammonia spirit, in wherein said ammonia spirit, the mass percentage of ammonia is 0.075% ~ 0.168%;
2) concentrated after extracting, be between 5.0-6.0 by concentrated solution adjust ph, add pectase and protease by the 1%-2% (m/v) of concentrated solution volume respectively, place, add 15%-20% (v/v) chitosan-acetic acid solution again, leave standstill;
3) filtration step 2) described in herb liquid, adjust ph is 5.0-7.0, concentrated, obtains Radix Glycyrrhizae extract;
4) Milkvetch Root is soaked, concentrated after extracting;
5) respectively by the 1%-2% (m/v) of concentrated solution volume to step 4) add pectase and protease in the concentrated solution that obtains, place, then add 15%-20% (v/v) chitosan-acetic acid solution, leave standstill;
6) filtration step 5) described in herb liquid, adjust ph is 5.0-7.0, concentrated, obtains Radix Astragali extract;
7) by step 3) Radix Glycyrrhizae extract and the step 6 prepared) Radix Astragali extract prepared mixes, and add water, adjust ph is to 4.0-6.0, centrifugal, and filter, fill, sterilizing, to obtain final product.
In the chitosan-acetic acid solution that the present invention is used, the mass concentration of chitosan is 1%.Its preparation method, such as getting chitosan, dissolving with 1% glacial acetic acid, making the solution that concentration is 1%.
Present invention also offers the product prepared by above-mentioned preparation method.
The present invention also provides according to the application of the Radix Astragali Radix Glycyrrhizae powder prepared by the inventive method preparation in preparation prevention infectious bronchitis of chicken medicine.
The invention provides preparation method of a kind of Radix Astragali Radix Glycyrrhizae powder and products thereof, have following advantage:
1, use the ammonia spirit of especially 0.3-0.6% (g/g) to extract Radix Glycyrrhizae, both fully can extract Angelica Polysaccharide, and significantly improve again the extraction ratio of glycyrrhizic acid.
2, adopt decompression low temperature reflux to extract, avoid high temperature to the destruction of active ingredient of Chinese herbs, remain active component to greatest extent.
3, protease and pectase is adopted to decompose the protein in concentrated solution and pectin, carry out flocculation with chitosan to concentrated solution to purify, effectively can remove the impurity components such as deproteinize, pectin, tannin, starch, retain the effective ingredient such as polysaccharide, glycyrrhizic acid, the powder clarity of preparation is higher, stability is better, more remarkable effect.
4, not high to equipment requirements, preparation technology is simple, can enhance productivity.
Detailed description of the invention
Material used and the source of equipment and lot number as follows:
The Radix Astragali, the place of production: Inner Mongol, lot number: 20110903, source: Henan thousand side pharmaceutcal corporation, Ltd;
Radix Glycyrrhizae, the place of production: Inner Mongol, lot number: 20111201, source: Henan thousand side pharmaceutcal corporation, Ltd;
Ammonia, analytical pure, lot number: 20100601, producer: Yantai City is Chemical Co., Ltd. in pairs;
Hydrochloric acid, analytical pure, lot number: 110301, producer: Luoyang chemical reagent factory;
Pectase, specification: 30,000 U/g, producer: Ningxia jade of the He family Bioisystech Co., Ltd;
Protease, specification: 100,000 U/g, producer: Ningxia jade of the He family Bioisystech Co., Ltd;
Chitosan, specification: deacetylation 85.5%, producer: Jinan Haidebei Marine Organism Engineering Co., Ltd.;
Extract at low temperature concentrates unit, model: TN-2/500, producer: Zhejiang Kaidi Pharmaceutical Chemical Machinery Co., Ltd.;
High speed centrifuge, model: GQLB-105N, producer: Liaoyang prosperous sunlight fluid separation device company limited;
Ultraviolet-visible spectrophotometer, model UV-2100, producer: Beijing Rayleigh Analytical Instrument Co., Ltd;
High performance liquid chromatograph, model e2695, UV-detector, model 2489, producer: water generation company of the U.S..
The specific embodiment of the present invention is further illustrated below by way of experimental example:
Embodiment 1: the preparation of Radix Glycyrrhizae extract
Take Radix Glycyrrhizae 50kg, be prepared by the following step:
1) extracting liquorice decoction pieces, adds in traditional Chinese herbs by thermal reflux low-temperature extracting tank, adds the purified water of medical material weight 8 times, then adds ammonia by 0.5% (v/v) of amount of water, soaks 30min; Control vacuum is-0.05MPa, at 70 DEG C, extracts 3 times, each 2h, extracting solution 80 eye mesh screens filter, and merging filtrate is evacuated in concentration tank, at 70 DEG C, being evaporated to relative density is 1.10 (70 DEG C), by dilute hydrochloric acid adjust ph to 5.0;
2) respectively by step 1) 2% (m/v) of concentrated solution volume add pectase and protease, stir, 4h is placed in 50 DEG C of insulations, then 1% (g/ml) chitosan-acetic acid solution is added by 15% (v/v) of concentrated solution volume, stir, leave standstill 12h;
3) by step 2) medical filtration, filtrate is by sodium hydroxide test solution adjust ph to 5.0, and at 70 DEG C, being evaporated to relative density is 1.20 (70 DEG C), is drying to obtain Radix Glycyrrhizae extract.
Embodiment 2: the preparation of Radix Astragali extract
Take Radix Astragali 50kg, be prepared by the following step:
1) get Radix Astragali decoction pieces, add in traditional Chinese herbs by thermal reflux low-temperature extracting tank, add the purified water of medical material weight 8 times, soak 30min; Control vacuum is-0.05MPa, and at 70 DEG C, extract 3 times, each 2h, extracting solution 80 eye mesh screens filter, and merging filtrate is evacuated in concentration tank, and at 70 DEG C, being evaporated to relative density is 1.10 (70 DEG C);
2) respectively by step 1) 2% (m/v) of concentrated solution volume add pectase and protease, stir, 4h is placed in 50 DEG C of insulations, then 1% (g/ml) chitosan-acetic acid solution is added by 15% (v/v) of concentrated solution volume, stir, leave standstill 12h;
3) by step 2) medical filtration, filtrate is by sodium hydroxide test solution adjust ph to 5.0, and at 70 DEG C, being evaporated to relative density is 1.20 (70 DEG C), is drying to obtain Radix Astragali extract.
Embodiment 3: the preparation of Radix Astragali Radix Glycyrrhizae powder
Radix Glycyrrhizae extract (being equivalent to crude drug in whole 25kg) prepared by Example 1, Radix Astragali extract (being equivalent to crude drug in whole 25kg) prepared by embodiment 2, mixes and obtains Radix Astragali Radix Glycyrrhizae powder.
Embodiment 4: the inventive method compares with other preparation methoies
Sample 1: get the Radix Astragali Radix Glycyrrhizae powder that the embodiment of the present invention 3 prepares gained.
Sample 2: adopt traditional aqueous extraction-alcohol precipitation technology to prepare Radix Astragali Radix Glycyrrhizae powder sample, its method is: take Radix Astragali decoction pieces and each 500g of licorice piece, put in round-bottomed flask, add the water soaking 1h of medical material gross weight 8 times amount, heating and refluxing extraction 2h, extracting solution 80 eye mesh screens filter, filtering residue adds 8 times of water gagings reflux, extract, 2 times again, each 2h, and gradation is filtered, merging filtrate, dry.
Sample 3: sample 4: adopt traditional aqueous extraction-alcohol precipitation technology to prepare Radix Astragali Radix Glycyrrhizae powder sample, its method is: take Radix Astragali decoction pieces and each 500g of licorice piece, put in round-bottomed flask, add the water soaking 1h of medical material gross weight 8 times amount, heating and refluxing extraction 2h, extracting solution 80 eye mesh screens filter, filtering residue adds 8 times of water gagings reflux, extract, 2 times again, each 2h, gradation is filtered, merging filtrate, relative density 1.20 (70 DEG C) is evaporated at 70 DEG C, add ethanol and reach 60% to alcohol content, stir, cold preservation 24h, filter, decompression filtrate recycling ethanol is concentrated into relative density 1.15 (70 DEG C), dry.
Study on the stability: the sample that above 3 kinds of methods are obtained, 100ml/ bottle, 40 DEG C, place 6 months under relative humidity 75% condition, character, pH value, glycyrrhizic acid content and total sugar content are investigated, the results are shown in Table 1.PH value algoscopy is carried out with reference to " People's Republic of China's veterinary drug allusion quotation " version in 2010 two annex 49 pages of pH value assay methods, and the content assaying method of glycyrrhizic acid and total sugar is as follows:
Glycyrrhizic acid measures according to high performance liquid chromatography (People's Republic of China's veterinary drug allusion quotation version in 2010 two annex 35 pages).
Chromatographic condition and system suitability take octadecylsilane chemically bonded silica as filler; With methanol-0.2mol/L Spirit of Mindererus .-glacial acetic acid (67:33:1) for mobile phase; Determined wavelength is 250nm.Number of theoretical plate calculates should be not less than 2000 by glycyrrhizic acid peak.
The extracting liquorice acid ammonium reference substance of preparing of reference substance solution is about 10mg, accurately weighed, puts in 50ml measuring bottle, dissolves and be diluted to scale, shaking up, to obtain final product with mobile phase.
The preparation of need testing solution takes this product 0.1g, accurately weighed, puts in 50ml measuring bottle, adds flowing and to make an appointment 20ml, supersound process (power 200W, frequency 50kHz) 30 minutes, takes out, lets cool, add mobile phase and be diluted to scale, shake up, to obtain final product.
Algoscopy is accurate respectively draws reference substance solution and each 10 μ l of need testing solution, injection liquid chromatography, measures, to obtain final product.
Total sugar measures according to ultraviolet visible spectrophotometry (People's Republic of China's veterinary drug allusion quotation version in 2010 two appendix 26 pages).
The preparation of reference substance solution gets anhydrous glucose reference substance in right amount, accurately weighed, adds water and makes the solution of every 1ml containing anhydrous glucose 30 μ g, to obtain final product.
It is accurately weighed that the preparation of need testing solution takes this product 0.1g, and put in 200ml measuring bottle, be diluted with water to scale, shake up, precision measures 5ml, puts in 200ml measuring bottle, is diluted with water to scale, shake up, to obtain final product.
Algoscopy precision measures reference substance solution and each 2ml of need testing solution, and put in tool plug test tube, precision adds 5% phenol solution 1ml, shake up, rapid precision adds sulphuric acid 5ml, after mixing of jumping a queue immediately, put in water-bath and be incubated 15min, take out, put in ice-water bath and cool 5min, take out, take corresponding reagent as blank, according to ultraviolet visible spectrophotometry (appendix 26 page), at 490nm wavelength, place measures absorbance respectively, calculate, to obtain final product.
Table 1 Radix Astragali of the present invention Radix Glycyrrhizae powder compares with the preparation stability that traditional preparation methods obtains (wherein assay data have been converted as dose in identical)
Conclusion: Radix Astragali Radix Glycyrrhizae powder prepared by the present invention adopts ammonia spirit to extract Radix Glycyrrhizae, and glycyrrhizic acid content significantly improves; Use protease and pectase and chitosan to carry out purification to concentrated solution, can the impurity component such as deproteinize, pectin be removed, effectively retain the effective ingredient such as polysaccharide, improve the stability of sample.
Embodiment 5 Radix Astragali Radix Glycyrrhizae powder is to the therapeutic test of artificial challenge's avian infectious bronchitis virus (IBV)
1 material
1.1 standard strain
Strain: infectious bronchitis virus Horte strain (bacterium numbering: CVCC-AV9), poison is purchased from China Veterinery Drug Inspection Office.
1.2 tests and contrast medicine
Radix Astragali Radix Glycyrrhizae powder prepared by embodiment 3; Silver yellow plate sticks up loose: 500g/ bag, and by Zhengzhou, Fu Yuan animal pharmaceutical estate company limited provides.
1.3 experimental animal
The SPF level chicken 90 of 21 ages in days.
1.4 key instrument
JA2003 type electronic balance: upper marine level device company limited; Within DT1000 days, inspire confidence in board electronic balance: Jin Yang counterweight Instrument Ltd. of Changshu City; Isolator.TGL-16G table model high speed centrifuge.
2 test methods
2.1 EID50 and counteracting toxic substances dosimetry
The rejuvenation of test strain adopts allantoic cavity inocalation method, and infectious bronchitis virus (IBV) is inoculated SPF Embryo Gallus domesticus, and Regular allantoic fluid is also preserved.
The Embryo Gallus domesticus median infective dose (EID50) of virus measures: in order to determine Chickens Infected CVCC-AV9, and the dosage of performance clinical symptoms, Embryo Gallus domesticus carries out the median infective dose determination test of IBV CVCC-AV9.The normal saline of the Embryo Gallus domesticus virus liquid mass fraction 0.9% of rejuvenation is carried out 10 times of serial dilutions, is respectively 10-4,10-5,10-6,10-7,10-8, each dilution factor inoculates 5 pieces of instar chicken embryos on the 10th, every embryonic breeding kind virus liquid 0.1mI.Establish saline control simultaneously.After the inoculation of paraffin sealing of hole, continue to cultivate observation 1 week in 37 DEG C, every 8h shines egg 1 time, the death condition of record Embryo Gallus domesticus.If chicken embryo death or blood vessel die back, idiosome are curled and motionlessly can be judged to be dead embryo; Correspondingly, Embryo Gallus domesticus increases naturally, blood vessel increases naturally is then slightly survival embryo.The median infective dose of chicken is calculated according to result.
The SPF chicken of random selecting 20 21 ages in days, be divided into 4 groups, often organize 5, when 25 age in days, allantois stock solution is carried out 10 times, 100,1000 times dilutions, respectively through trachea instillation allantois stock solution and 10 times, 100 times, the 1000 times allantoic fluid 0.2mL diluted thereof, observe the mental status of chicken after counteracting toxic substances every day, drinking-water situation of searching for food, there is slight cough, sneeze, rhinorrhea, dyspnea, dehisce to pant, arrange white feces, the chicken of the dry a certain symptom of toe is judged to morbidity, cause the dosage of 80% chicken mass-sending disease as counteracting toxic substances dosage.
2.2 animal grouping and process
70 25 age in days SPF chickling are divided into 7 groups at random, often organize 10.Radix Astragali Radix Glycyrrhizae powder treatment group (1-4 group), silver yellow plate stick up loose matched group (the 5th group), positive controls (the 6th group), negative control group (the 7th group), 1-6 group inoculates IBV suspension (80% chicken morbidity dosage, determines according to challenge test result) in 25 ages in days through trachea.Radix Astragali Radix Glycyrrhizae treatment group and positive drug control group, occur that clinical onset symptom starts administration, clinical symptoms shows as appearance slight cough, sneeze, rhinorrhea, dyspnea, dehisces to pant, arrange white feces, diarrhoea, the dry middle any one symptom of toe.Administration concentration computational methods: first 3 days of administration, measure the amount of drinking water that one day 24h often organizes chicken group, and the amount of drinking water average out to 52ml/ of chicken only.Drug level is calculated according to amount of drinking water and dosage.Drug level (mg/mL)=52ml/ × dosage.The warm water of various dose group same volume dissolves, and the amount of gavaging of every chicken is 1ml, and test control group gavages 1ml purified water.Every day 1 time, be used in conjunction 7 days.Gavage method: Baoding upward, Caput Gallus domesticus portion, with the 5ml syringe of white rubber tube being connected with 5.5cm, after inserting bottleneck throat, along with the swallowing act of chicken slowly inserts esophagus, insert 2/3 of rubber tube, inject medicinal liquid.Isolator is raised, and conveniently manages.Day by day the mental status of observed and recorded chicken and clinical manifestation, in table 2.
Table 2 is group test processing method respectively
2.3 judge index
2.3.1 clinical symptoms and pathological change
2.3.2 sickness rate, mortality rate, cure rate is calculated
(1) sickness rate
Allly there is slight cough, sneeze, rhinorrhea, dyspnea at duration of test, dehisce to pant, arrange any one infectious bronchitis (IB) the classical symptom persons such as white feces, toe be dry and be morbidity, add up with this and fall ill chicken number and calculate sickness rate.
Sickness rate=morbidity chicken number/this group test chicken sum × 100%.
Mortality rate
Allly there is above-mentioned IB classical symptom and dead at duration of test, and the visible trachea of necropsy, nasal cavity have serosity, Catarrhal and caseous secretions, there is the IB typical pathologic changes such as piebaldism kidney, renal swelling, and differentiate, for positive is judged to infect death, to add up and die of illness chicken number and calculate mortality rate through PCR pathogen separation.
Mortality rate=infect dead chicken number/this group for examination chicken sum × 100%.
Cure rate
Duration of test, the sick chicken for the treatment of group after administration without slight cough, sneeze, rhinorrhea, dyspnea, dehisce to pant, arrange the symptoms such as white feces, toe be dry, and when the mental status and appetite are recovered normal, be judged to cure, calculating cure rate.
Cure rate=cure disease chicken number/this group morbidity chicken sum × 100%.
2.3.3 the mensuration of Immune Organs Index
After medication when 15 days, cut open and kill residue chicken, win spleen, fabricius bursa, filter paper sucks the moisture on surface, weighs with analytical balance, Computation immunity shoot formation (the heavy mg/ body weight g of Immune Organs Index=immune organ)
3 statistical procedures
Experimental data represents with mean ± standard deviation, and adopt SPSS18.0 statistical software to analyze, compare between group with one factor analysis of variance, 0.01<P≤0.05 is significant difference, and P≤0.01 is that difference is extremely remarkable.
4 result of the tests
4.1 viral EID50 measurement results
Cut open inspection Embryo Gallus domesticus when the dead Embryo Gallus domesticus of criterion: 24-144h and 144h to occur obviously rolling up, the Embryo Gallus domesticus of the IB typical cytopathics such as dehydration is judged to infection.
The pathological changes of 10-7 dilution is higher than 50%; The pathological changes of 10-8 dilution is lower than 50%; 50% pathological changes is divided
IgEID50=higher than the logarithm (10 times go forward one by one dilution be 1) of the low dilution factor logarithm-respective distance ratio × coefficient of dilution of 50% pathological changes, thus tries to achieve EID50.
Calculate according to Reed-Muench method, the EID50 of IgEID50=-7-0.33 × 1=-7.33, IBV-Horte strain on Embryo Gallus domesticus is 10-7.33/0.1ml.Each group of embryo pathological changes situation is in table 5.
Table 3 IBV-Horte strain EID50 measurement result
4.2 counteracting toxic substances dose measurements
After counteracting toxic substances, 24h starts morbidity, virus allantois liquid inductance dye group chicken has 4 morbidities, show as mouth breathing, sneeze, trachea rale, discharge white loose stool, shake all over, wherein have 2 death, dissect in visible trachea-bronchial epithelial cell and have mucus, kidney enlargement, has the typical chicken Kidney infectious bronchitis symptoms such as urate deposition.10 times of diluent group chickens have 3 chickens to occur trachea rale, arrange white feces.100 times of dilutions and 1000 times of virus dilution groups, only have 1 to occur dyspnea, slight mouth breathing.Determine that best counteracting toxic substances dosage is virus stock solution used trachea inoculation 0.2ml.
4.3 clinical symptoms and pathological change
After administration, the overall mental status of the chicken group of medication group is good, amount of drinking water and feed intake normal, only have indivedual chicken to show spirit depressed.And positive controls chicken group shows and occurs that spirit is depressed, amount of drinking water of searching for food declines, even useless absolutely, there is Calx sample feces in ataxia, in process of the test, mortality rate is 30%.Negative control group isolated rearing, drinking-water of searching for food is normal, without clinical change.After medication 15 days, chicken was all cutd open inspection and finds, the kidney for the treatment of group chicken group is micro-swollen, but occurs without piebaldism kidney, and also without urate deposition in ureter, feces is normal.The swelling of positive controls chicken kidney, piebaldism kidney, tracheal strips have a small amount of mucus.
4.4 Radix Astragali Radix Glycyrrhizae powder are on the impact of the sickness rate of animal, mortality rate and cure rate
After counteracting toxic substances, the chicken mortality rate that the sickness rate of chicken to be the mortality rate of the Radix Astragali Radix Glycyrrhizae powder chicken of 70%-80%, 0.5g/L-1g/L be 0%, 0.25g/L Radix Astragali Radix Glycyrrhizae powder and silver yellow plate sticks up loose drug control group is 10%.In 0.25g/L-1g/L Radix Astragali Radix Glycyrrhizae powder, the cure rate of 4 dosage groups is respectively 75%, 87.5%, 100%, 87.5%, and it is 75% that drug control group silver yellow plate sticks up loose cure rate.Its effect of Radix Astragali Radix Glycyrrhizae powder of 0.5g/L is better than silver yellow plate and sticks up loose.In table 6.
Table 4 Radix Astragali Radix Glycyrrhizae powder is on the impact of the sickness rate of animal, mortality rate and cure rate
4.5 Radix Astragali Radix Glycyrrhizae powder is on the impact of Immune Organs of Chicken index
The Radix Astragali Radix Glycyrrhizae powder of 0.5-1g/L, silver yellow plate stick up loose compared with positive controls, improve thymus index (P<0.05) significantly, the Radix Astragali Radix Glycyrrhizae powder of 0.5g/L with 1g/L improves index and spleen index (P<0.05) significantly compared with positive controls, the Radix Astragali Radix Glycyrrhizae powder of 0.25-1g/L, silver yellow plate stick up loose drug control group significant difference compared with positive controls, improve bursal index (P<0.05) significantly.In table 7.
Table 7 Radix Astragali Radix Glycyrrhizae is on the impact of Immune Organs of Chicken index
Note: " * " represents significant difference (0.01<P≤0.05) compared with positive controls, " * * " represents that difference extremely significantly (P≤0.01) compared with positive controls.
5 conclusions and discussion
This test selects SPF chickling as experimental subject.Although the equal susceptible IBV of chicken of bibliographical information institute has age, the infection morbidity of chickling is even more serious, and sickness rate reaches 100%, and mortality rate can up to more than 50%.After chicken more than 6 week age infects, respiratory symptom is then comparatively gentle, and rarely found death.Because strain is different, also have the factors such as individual variation, we adopt the virus stock solution used after rejuvenation to carry out counteracting toxic substances, and sickness rate reaches 80%.Successfully establish avian infectious bronchus model.
After gavaging Radix Astragali Radix Glycyrrhizae powder, clinical symptom disappearance, the Radix Astragali Radix Glycyrrhizae powder cure rate of 0.5g/L reaches 87.5%, curative effect be significantly better than that silver yellow plate sticks up loose drug control group 75%.Meanwhile, the Radix Astragali Radix Glycyrrhizae powder of various dose can improve thymus index, index and spleen index and bursal index, thus the resistance of enhancing body.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (10)
1. a preparation method for Radix Astragali Radix Glycyrrhizae powder, comprises the following steps:
S1, prepares Radix Astragali extract with Milkvetch Root;
S2, prepares Radix Glycyrrhizae extract with licorice medicinal materials;
Described Radix Astragali extract and described Radix Glycyrrhizae extract mixed preparing are Radix Astragali Radix Glycyrrhizae powder by S3.
2. preparation method according to claim 1, is characterized in that, the preparation process of described Radix Astragali extract comprises:
A) Milkvetch Root is soaked, concentrated after extracting;
B) concentrated solution step a) obtained first processes with pectase and protease, then processes with chitosan-acetic acid solution;
C) by step b) mixture that obtains filters, and filtrate is adjusted to faintly acid to neutral, being preferably adjusted to pH value is 5.0-7.0, then concentrates, dry, obtains Radix Astragali extract.
3. preparation method according to claim 2, is characterized in that, in step b) in, the consumption of pectase and protease is 1-2g/100mL concentrated solution.
4. the preparation method according to Claims 2 or 3, is characterized in that, the mass ratio of pectase and protease is 1:2-2:1.
5. the preparation method according to any one of claim 2-4, is characterized in that, step b) in, the addition of chitosan-acetic acid solution and the volume ratio of concentrated solution are 0.15:1-0.2:1.
6. the preparation method according to any one of claim 1-5, is characterized in that, the preparation process of described Radix Glycyrrhizae extract comprises:
A) licorice medicinal materials is soaked with ammonia spirit, concentrated after extracting;
B) by steps A) the concentrated solution adjust ph that obtains is 5.0-6.0, then first processes with pectase and protease, then processes with chitosan-acetic acid solution;
C) by step B) mixture of gained filters, and filtrate is transferred to faintly acid to neutral, preferred adjust ph is 5.0-7.0, concentrated, dry, obtains Radix Glycyrrhizae extract.
7. the preparation method according to any one of claim 1-6, is characterized in that, steps A) in, in described ammonia spirit, the mass content of ammonia is 0.075% ~ 0.168%.
8. the preparation method according to claim 6 or 7, is characterized in that, step B) in, the consumption of pectase and protease is 1-2g/100mL concentrated solution.
9. the preparation method according to any one of claim 6-8, is characterized in that, step B) in, the addition of chitosan-acetic acid solution and the volume ratio of concentrated solution are 0.15:1-0.2:1.
10. the application of the Radix Astragali Radix Glycyrrhizae powder prepared by any one of claim 1-9 preparation method in preparation prevention infectious bronchitis of chicken medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510256517.1A CN104983777A (en) | 2015-05-19 | 2015-05-19 | Preparation method for radix astragali and licorice powder and product thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510256517.1A CN104983777A (en) | 2015-05-19 | 2015-05-19 | Preparation method for radix astragali and licorice powder and product thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104983777A true CN104983777A (en) | 2015-10-21 |
Family
ID=54295766
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510256517.1A Pending CN104983777A (en) | 2015-05-19 | 2015-05-19 | Preparation method for radix astragali and licorice powder and product thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104983777A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108524632A (en) * | 2018-04-11 | 2018-09-14 | 哈尔滨市康隆药业有限责任公司 | A kind of preparation process of innovation specification concentrated type oral liquid for cough and asthma of children |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104208128A (en) * | 2013-05-30 | 2014-12-17 | 洛阳惠中兽药有限公司 | Traditional Chinese medicinal composition and preparation method and application |
-
2015
- 2015-05-19 CN CN201510256517.1A patent/CN104983777A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104208128A (en) * | 2013-05-30 | 2014-12-17 | 洛阳惠中兽药有限公司 | Traditional Chinese medicinal composition and preparation method and application |
Non-Patent Citations (1)
| Title |
|---|
| 赵冰俞等: "兽用黄芪多糖注射液生产新技术研究", 《河南科技》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108524632A (en) * | 2018-04-11 | 2018-09-14 | 哈尔滨市康隆药业有限责任公司 | A kind of preparation process of innovation specification concentrated type oral liquid for cough and asthma of children |
| CN108524632B (en) * | 2018-04-11 | 2020-11-06 | 哈尔滨市康隆药业有限责任公司 | Preparation process of concentrated oral liquid for treating infantile cough and asthma |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR20110132407A (en) | Plant extract compositions for prevention and treatment of influenza | |
| CN101780153B (en) | Traditional Chinese drug composition for pig and preparation method and application thereof | |
| CN101596246B (en) | Smoked plum extractive and wild jujube seed extractive compound preparation as well as preparation method and application thereof | |
| CN102716180A (en) | Right supporting and toxin eliminating oral liquid and preparation method of oral liquid | |
| CN101869595A (en) | Preparation method and quality detection method of liquorice and gall oral liquid | |
| CN103784538A (en) | Traditional Chinese medicinal compound drug for effectively preventing and treating swine influenza | |
| CN102670694B (en) | Traditional Chinese medicine combination for treating porcine pseudorabies and preparation method and application thereof | |
| CN103638147A (en) | Traditional Chinese medicine compound for resisting swine influenza virus | |
| CN104288217A (en) | Traditional Chinese medicine composition and application thereof | |
| CN106389562A (en) | Pudilan Xiaoyan granules, preparation method thereof, and product quality control method | |
| CN104983777A (en) | Preparation method for radix astragali and licorice powder and product thereof | |
| CN105079143A (en) | Medicine composition for treating nephropathy | |
| CN108553504A (en) | Join stilbene powder and preparation method thereof | |
| CN103463244A (en) | Method for extracting blood sugar lowering substance from China roses and application of blood sugar lowering substance | |
| CN106237050A (en) | A kind of Chinese medicine compound of effective preventing and treating H1N1 and H3N2 swine flue and pig blue-ear disease and preparation method thereof | |
| CN103655757A (en) | Traditional Chinese medicine compound for effectively resisting swine influenza virus | |
| CN106491957B (en) | It is a kind of for treating the preparation method of the Chinese medicine composition of bronchial asthma | |
| CN105816583B (en) | Compound medicine for treating cold and preparation method thereof | |
| CN104784327A (en) | Traditional Chinese veterinary medicine composition for treating respiratory diseases of chicken and preparation method thereof | |
| CN105434895A (en) | Gastrodia elata rhodiola rosea cream and preparation method thereof | |
| CN113181229B (en) | Application of cabbage type rape-isatis tinctoria G monomer addition system in inhibiting novel coronavirus SARS-CoV-2 | |
| CN103271983A (en) | Perfusate for treating endometritis and kysthitis of pig as well as preparation method and application thereof | |
| CN106421329A (en) | Traditional Chinese medicine compound prescription for simultaneously resisting swine infuenza and porcine reproductive and respiratory syndrome and extraction method thereof | |
| CN103356813B (en) | Indian stringbush root capsule | |
| CN102266383B (en) | Application of Mesona chinensis Benth in preparing medicines for treating viral infectious diseases in livestock and poultry |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20151021 |