CN101926861A - Method for preparing Chinese medicinal veterinary medicament for enhancing non-specific immune function - Google Patents
Method for preparing Chinese medicinal veterinary medicament for enhancing non-specific immune function Download PDFInfo
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Abstract
The invention relates to a veterinary Chinese medicinal composition for improving growth performance and enhancing the non-specific immune function of an organism. The composition is prepared from the following raw material medicaments: Chinese pulsatilla root, astragalus, large head atractylodes rhizome, indigowoad root, baical skullcap root, bupleurum and rhubarb. The invention also relates to a veterinary medicament prepared from the veterinary Chinese medicinal composition, a method for preparing the veterinary medicament and the application of the veterinary Chinese medicinal composition. The veterinary Chinese medicinal composition has the effects of strengthening body resistance, eliminating evil, clearing away heat and toxic material and enhancing the non-specific immune function of the organism and can treat various atypical symptoms such as low immune function of the organism, immune tolerance, endogenous toxin, stress and the like, the decrease of the growth performance and damp-heat symptoms such as anemopyretic cold, flu, gumboro disease, enterotoxin syndrome and the like.
Description
Technical field
The present invention relates to field of veterinary, particularly, the present invention relates to a kind of Chinese medicine composition for animals that is used to improve growth performance, enhancing body non-specific immunity, and veterinary medicine, the preparation method of this veterinary medicine and the application of this Chinese medicine composition for animals of making by this Chinese medicine composition for animals.
Background technology
Birds is important economic animal, and the epiornitic disease is restricting the development of aviculture always.Although there are a lot of bird vaccines can be for for selecting at present, they be at specific immunity mostly, and onset is slow, act ons single-mindedly, and viral infection has just taken place before vaccine provides effective protection birds sometimes.Nonspecific immunity be the opposing infected by microbes the first line of defence, have rapid-action, the characteristics that sphere of action is wide.Therefore, the novel immunomodulator that can effectively activate the birds nonspecific immunity is being sought and identified to Many researchers all at present.
Some drugs in the natural Chinese medicine and extract thereof, have multiple effects such as the animal body of enhancing immunity, promotion growth, inhibition pathogen, and utilize the Chinese herb prevention Animal diseases to meet the trend of green cultivation, many large-scale plants also more and more pay attention to the enhance immunity effect of Chinese medicine health care product at present.Thereby research Chinese medicine just seems rather important to the influence of birds non-specific immunity and growth performance thereof, has certain Research Significance and using value.
The inventor finds after consulting pertinent literature, at home and abroad there is no at present and can effectively improve the birds growth performance, strengthens the Chinese medicine composition for animals of birds body non-specific immunity.
Summary of the invention
In order to realize purpose of the present invention, the invention provides a kind of Chinese medicine composition for animals, it is made by following crude drug: the Radix Pulsatillae, the Radix Astragali, the Rhizoma Atractylodis Macrocephalae, Radix Isatidis, Radix Scutellariae, Radix Bupleuri, Radix Et Rhizoma Rhei.This Chinese medicine composition for animals can be used in and improves growth performance, enhancing body non-specific immunity.
In described Chinese medicine composition for animals, the preferred weight portion proportioning of each crude drug component is: the Radix Pulsatillae 10～30 weight portions, the Radix Astragali 10～30 weight portions, the Rhizoma Atractylodis Macrocephalae 5～15 weight portions, Radix Isatidis 5～15 weight portions, Radix Scutellariae 5～15 weight portions, Radix Bupleuri 5～15 weight portions, Radix Et Rhizoma Rhei 5～15 weight portions; The preferred weight portion proportioning of each crude drug component is: the Radix Pulsatillae 20～30 weight portions, the Radix Astragali 20～30 weight portions, the Rhizoma Atractylodis Macrocephalae 10～15 weight portions, Radix Isatidis 10～15 weight portions, Radix Scutellariae 10～15 weight portions, Radix Bupleuri 10～15 weight portions, Radix Et Rhizoma Rhei 5～10 weight portions; The most preferred weight portion proportioning of each crude drug component is: the Radix Pulsatillae 25 weight portions, the Radix Astragali 25 weight portions, the Rhizoma Atractylodis Macrocephalae 10 weight portions, Radix Isatidis 10 weight portions, Radix Scutellariae 10 weight portions, Radix Bupleuri 10 weight portions, Radix Et Rhizoma Rhei 10 weight portions.
The present invention also provides a kind of veterinary medicine that is used to improve growth performance, enhancing body non-specific immunity, and it is made by described Chinese medicine composition for animals and pharmacy acceptable auxiliary.The dosage form of described veterinary medicine includes but not limited to oral solution, granule, soluble powder, pre-mixing agent.The adjuvant commonly used that wherein used pharmacy acceptable auxiliary is known for those skilled in the art, the conventional method that the preparation method of veterinary medicine is known for those skilled in the art, specifically can be referring to " veterinary drug preparation production application handbook (Jiangxi science tech publishing house, in December, 1994 publication), " pharmaceutic adjuvant complete works " (Sichuan science tech publishing house, publication in January nineteen ninety-five), " Chinese veterinary drug allusion quotation " two ones of versions (Chinese agriculture publishing house, publish in April, 2006) in 2005.
The present invention also provides a kind of method for preparing described veterinary medicine, and this method comprises:
1) the weighting raw materials Radix Pulsatillae, the Radix Astragali, the Rhizoma Atractylodis Macrocephalae, Radix Isatidis, Radix Scutellariae, Radix Bupleuri, Radix Et Rhizoma Rhei.
2) Radix Pulsatillae, the Radix Astragali, Radix Isatidis, Radix Scutellariae, Radix Et Rhizoma Rhei are pulverized, add the decocting that 3-8 doubly measures and boiled 1-4.5 hour, the leaching decocting liquid is standby.Wherein pulverising step preferably is ground into coarse powder with crude drug; Decocting step preferably divides and carry out for 3 times: the decocting that adds 8 times of amounts of medical material for the 1st time boiled 3 hours, and the decocting that adds 5 times of amounts of medical material for the 2nd time boiled 2 hours, and the decocting that adds 3 times of amounts of medical material for the 3rd time boiled 1 hour.
3) Rhizoma Atractylodis Macrocephalae, Radix Bupleuri are pulverized, added the decocting that 3-8 doubly measures and boiled 1-3 hour, it is standby to collect volatile oil, and the leaching decocting liquid is standby.Wherein pulverising step preferably is ground into coarse powder with crude drug; Decoct step and preferably crude drug is soaked after adding water, before decocting, wherein soak time is preferably 45 minutes; Amount of water is preferably the water of 8 times of amounts of medical material, and decocting time is preferably 3 hours.
4) with filter in the step 3) medicinal residues add the decocting that 3-8 doubly measures and boiled 1-3 hour, the leaching decocting liquid merges standby with the decocting liquid of gained in the step 3).Wherein decocting step preferably divides and carry out for 2 times: the decocting that adds 5 times of amounts of medical material for the 1st time boiled 2 hours, and the decocting that adds 3 times of amounts of medical material for the 2nd time boiled 1 hour.
5) with step 2) in gained decocting liquid and the step 4) collecting decoction of gained merge, amalgamation liquid is concentrated, make every 1ml concentrated solution be equivalent to contain the 0.5g crude drug; Add ethanol then in concentrated solution, regulating pH value is 5.0～7.5, leaves standstill, and gets supernatant.Wherein add alcoholic acid amount and be preferably that to make the ethanol final concentration be 60% amount; The reagent of regulating pH value is preferably 10%NaOH solution.
6) with behind the supernatant decompression recycling ethanol of gained in the step 5) with step 3) in the volatile oil of gained merge, drying is made extract powder, and is standby.
7) extract powder that step 6) made adds the pharmacy acceptable auxiliary and makes described veterinary medicine, for example oral solution, granule, soluble powder, pre-mixing agent, and make every 1g or every 1ml veterinary medicine be equivalent to contain the 1g crude drug.
The present invention also provides a kind of described Chinese medicine composition for animals to be used for improving the application of the veterinary medicine of growth performance, enhancing body non-specific immunity in preparation.
The present invention's Chinese medicine composition for animals, veterinary medicine are specially adapted to poultry, include but not limited to chicken, duck, goose.
Veterinary medicine of the present invention can be by admixing feedstuff or being blended into the mode administration of drinking-water, makes animal pass through free choice feeding or freely drink water and take in veterinary medicine of the present invention.The drug level of admixing feedstuff is 0.5% (W/W), and promptly every 100g feedstuff is admixed 0.5g veterinary medicine of the present invention; The drug level that is blended into drinking-water is 0.3-0.5% (W/W), and promptly every 100g drinking water is blended into 0.3-0.5g veterinary medicine of the present invention.Administration number of times is for once a day, successive administration 5 to 7 days.
The thin layer chromatography that two appendix of veterinary medicine employing " Chinese veterinary drug allusion quotation " version in 2005 of the present invention are 31 pages is carried out baicalin, baicalin, Radix Pulsatillae extract and the Radix Et Rhizoma Rhei extract in the described veterinary medicine of qualitative determination; With reference to the astragalus polysaccharides in " veterinary drug quality standard " described veterinary medicine of version qualitative determination in 2003; The employing reversed phase high-performance liquid chromatography (referring to Wang Qian, etc., the content of anemoside B4 in the rp-hplc determination Radix Pulsatillae medical material, " time precious traditional Chinese medical science traditional Chinese medicines ", in November, 2009, the 20th volume o. 11th), the content of anemoside B4 in the mensuration medicine.
1) qualitative determination of baicalin, baicalin:
Get veterinary medicine 1g of the present invention, add ethyl acetate-methanol (3: 1) mixed solution 30mL, reflux 30 minutes is put coldly, filters, and filtrate evaporate to dryness, residue add methanol 5mL makes its dissolving, gets supernatant as need testing solution.Other gets baicalin reference substance, baicalin reference substance, adds methanol and makes the reference substance solution that every 1mL contains 1mg, 0.5mg respectively.Test according to thin layer chromatography (31 pages of appendix), draw need testing solution, each 1 μ L of above-mentioned two kinds of reference substance solution, put respectively on same polyamide film, with toluene-ethyl acetate-methanol-formic acid (10: 3: 1: 2) be developing solvent, presaturation 30 minutes, launch, take out, dry, put under the uviol lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show two identical dark spots.
(2) mensuration of Radix Pulsatillae extract:
Get veterinary medicine 4g of the present invention, add ethanol 20mL, reflux 1 hour, filter, filtrate is concentrated into about 6mL, puts cold, it is an amount of to add acetone, then generate precipitation, filter, speed is got precipitation a small amount of (about 5g), put in the test tube, add acetic anhydride 1mL and make dissolving, add sulphuric acid 1mL along tube wall, two liquid junction places show redness or purple colour circle.
(3) mensuration of Radix Et Rhizoma Rhei extract:
Get veterinary medicine 0.1g of the present invention, add methanol 20mL, soaked 1 hour, filter, get filtrate 5mL, evaporate to dryness, residue adds water 10mL makes dissolving, adds hydrochloric acid 1mL again, reflux 30 minutes, cooling immediately divides 2 joltings to extract with ether, each 20mL, merge ether solution, evaporate to dryness, residue add chloroform 1mL makes dissolving, as need testing solution.Other gets Radix Et Rhizoma Rhei control medicinal material 0.1g, shines medical material solution in pairs with legal system.Get the chrysophanic acid reference substance again, add methanol and make the solution that every 1mL contains 1mg, in contrast product solution.Test according to thin layer chromatography (31 pages of appendix), draw each 4 μ L of above-mentioned three kinds of solution, put respectively in same be on the silica gel H lamellae of adhesive with the sodium carboxymethyl cellulose, upper solution with petroleum ether (30-60 ℃)-Ethyl formate-formic acid (15: 5: 1) is developing solvent, launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with control medicinal material chromatograph relevant position on, show five identical orange-yellow fluorescence principal spots; With the corresponding position of reference substance chromatograph on, show identical orange-yellow fluorescence speckle, put in the ammonia steam smoked after, speckle becomes redness.
(4) mensuration of astragalus polysaccharides:
Get veterinary medicine 5g of the present invention, add deionized water 20mL, soaked 30 minutes, filter, get filtrate 5mL, add 3 of 2% alpha-Naphthol alcoholic solution, after shaking up, slowly add sulphuric acid 0.5mL, show brown ring at two liquid level intersections along test tube wall.
(5) content measurement of anemoside B4
Get veterinary medicine 1g of the present invention, accurately measure, place tool plug conical flask, the accurate then methanol 10ml that adds, ultrasonic 30 minutes, put coldly, to filter, filtrate is put in the 50ml measuring bottle, with a small amount of mobile phase (acetonitrile: 0.1% phosphoric acid (V: V; 28: 72)) washing container and residue, washing liquid is incorporated in the same measuring bottle, adds mobile phase and is diluted to scale, shakes up, and crosses 0.2 μ m filter membrane, detects through reversed-phase high-performance liquid chromatography, calculates, and the content of anemoside B4 (g/g) all must not be lower than 0.4%.
In the present invention's Chinese medicine composition for animals, the Radix Pulsatillae is the dry root of Ranunculaceae Radix Pulsatillae Pulsatilla chinensis (Bge.) Regel, mainly contains compositions such as pulchinenoside, triterpenic acid, lignanoid, Radix Pulsatillae spirit, Radix Pulsatillae English, Anemonin, protoanemonin, daucosterol and glycoprotein.Wherein pulchinenoside has effects such as antitumor, raise immunity, anti-ameba and sterilization.The Radix Pulsatillae and pulchinenoside have certain facilitation to chicken weightening finish, immune organ function, slightly be better than the Radix Astragali, the phagocytic rate of chicken neutrophilic granulocyte is had extremely significant potentiation, the Radix Pulsatillae and saponin thereof, as immunostimulant, have excellent research and be worth and application prospect.
The Radix Astragali is the dry root of leguminous plant Radix Astagali Astragalus membranaceus (Fisch.) Bge.var.mongholicus (Bge.) Hsiao or Radix Astragali Astragalus membranaceus (Fisch.) Bge., is generally believed the Chinese medicine simply that animal immune is had facilitation.The Radix Astragali can strengthen the phagocytic function of reticuloendothelial system, and leukocyte and multinuclear leucocyte quantity are significantly increased, and macrophage phagocytic percentage rate and phagocytic index are significantly risen, and humoral immunization, cellular immunization are all had facilitation.The Radix Astragali has the ability of enhanced virus inducement interferon.The easy catching a cold person takes the Radix Astragali in the flu epidemic season, times of common cold is obviously reduced, and cold symptoms is alleviated, and the course of disease shortens.The contained effective ingredient astragalus polysaccharides of the Radix Astragali has obvious anti-fatigue, low temperature resistant effect simultaneously, and the resisting fatigue time of normal and deficient mice is obviously prolonged.The Radix Astragali also can make the cells physiological metabolism strengthen, and can promote the protein of serum and liver to upgrade, and protein metabolism is had facilitation.The Radix Astragali all has inhibitory action to dysentery, Diplococcus pneumoniae, Hemolytic streptococcus A, B, C and golden yellow, lemon yellow, Staphylococcus albus etc.
The Rhizoma Atractylodis Macrocephalae is the dry rhizome of feverfew Rhizoma Atractylodis Macrocephalae Atractylodes macrocephala Koidz, has invigorating the spleen and benefiting QI, dampness diuretic, hidroschesis, antiabortive effect.Be used for insufficiency of the spleen lack of appetite, abdominal distention is had loose bowels, phlegm retention vertigo and palpitation, edema, spontaneous perspiration, frequent fetal movement etc.Rhizoma Atractylodis Macrocephalae decocting liquid has excitation to gastrointestinal motility, has antiulcer, and resisting fatigue can significantly strengthen the ability of leukocytes phagocytic staphylococcus aureus.The Rhizoma Atractylodis Macrocephalae can promote the mouse red blood cell hemoposieis simultaneously, can effectively suppress lipid peroxidation, reduces the content of organizing lipid peroxide, avoids the destruction of harmful substance to the histiocyte 26S Proteasome Structure and Function.The Rhizoma Atractylodis Macrocephalae has cytotoxicity to oncocyte, can reduce the rate of increase of oncocyte, lowers the aggressive of tumor tissue, improves the ability of body antitumor reaction.
Radix Isatidis is the dry root of Cruciferae Isatis indigotica Fort. Isatis indigotica Fort., have clearing away heat and cooling blood, antibiosis and antiviral functions clinically, its infusion is to bacillus subtilis, staphylococcus aureus, sarcine, escherichia coli, Bacillus typhi, paratyphoid fever bacillus, dysentery (will Hayes, Fu Shi) bacillus, bacillus enteritidis etc. all have inhibitory action, and Radix Isatidis has good Detoxication simultaneously.It is heavy that all Radix Isatidis polysaccharide that contain of Radix Isatidis can significantly increase the normal mouse spleen, and total white blood cells and lymphocyte number can also obviously strengthen antibody and form function, strengthen mouse mainline carbon clearance speed.Radix Isatidis ingredient indirubin can strengthen the phagocytic activity of the mononuclear phagocyte system of animal, and the human body immunity improving function has certain antitumaous effect.
Radix Scutellariae is the dry root of labiate Radix Scutellariae Scutellaria baicalensis Georgi.It has wider antimicrobial spectrum, dysentery bacterium, diphtheria corynebacterium, bacillus pyocyaneus, staphylococcus, streptococcus, Diplococcus pneumoniae and meningococcus etc. all there is inhibitory action in vitro, decoct is done the larynx spraying, also effective to the meningitis bacillicarrier, even the staphylococcus aureus to antibiotics such as penicillin have developed immunity to drugs still belongs to sensitivity to Radix Scutellariae.To the various skin pathogenic fungus, external also have the inhibition effect, and can kill leptospira.The contained baicalin of Radix Scutellariae, noroxylin have the antiinflammatory anti-allergy action.Baicalin has blood pressure lowering, diuresis, sedation, can suppress the spontaneous activity of white mice, action intensity is relevant with dosage, the Radix Scutellariae decoct can suppress the mice positive conditioned reflex, this effect may be because it has strengthened can be used for the hyperpietic that nerve excitability increases and has a sleepless night due to the cortex process of inhibition.The tincture intravenous injection of Radix Scutellariae rhizome to the frog, cat, the Canis familiaris L. of strychnism, can be eliminated the symptom of tonic spasm, and make animal avoid death.
Radix Bupleuri is the dry root of umbelliferae bupleurum Bupleurum chinense DC. or Radix Bupeuri Scorzonerfolii. Bupleurum scorzonerifolium Willd..By the character difference, practise title " Radix Bupleuri " respectively and reach " Radix Bupleuri Scorzonerifolii ".Its contained volatile oil (eugenol, caproic acid, r-undecanoic acid lactone and to the methoxybenzene diketone), saikoside (sapogenin A) has the trivial solution heat effect to heating due to typhoid fever, paratyphoid fever vaccine, escherichia coli liquid, fermented milk, the yeast etc., and the animal normal body temperature is descended.Radix Bupleuri total saponin, saponin also have calmness, analgesia, antitussive effect, and swollen and connective tissue proliferation inflammation all has inhibitory action to ankle joint due to the multiple proinflammatory agent.Saikogenin a and d, bupleurumol, hitodesterol can blood fat reducing (at experimental hyperlipidemia animal cholesterol reducing, triglyceride and phosphide on one's body), protect the liver (to CCl
4, hepatic insufficiency has protective effect due to ethanol, antityphoid vaccine, yolk, mouldy rice, D-galactosamine etc.Performance: the transaminase reduces, and liver function recovery, hepatic injury alleviate).Radix Bupleuri polysaccharide in the Radix Bupleuri can make phagocyte, natural killer cell increased functionality, improves the special viral antibody titre, and influenza virus, hepatitis virus, vaccinia virus, I type poliomyelitis virus, herpesvirus are had good inhibitory effect.
Radix Et Rhizoma Rhei is the dry root and rhizome of polygonum rheum palmatum Rheum palmatum L., Rheum tanguticum Rheum tanguticum Maxim.ex Balf. or Rheum officinale Rheum officinale Baill..Clinically be usually used in treating constipation due to dry stool, constipation with heat retention, but general 6-19 hour defecation after medication.Various experimental results show that: Radix Et Rhizoma Rhei has in various degree inhibitory action to multiple gram positive bacteria, some negative bacterium and fungus, and its bacteriostatic effective ingredient is an anthraquinone derivative, and wherein the effect of chrysophanic acid, emodin, aloe-emodin is the strongest.Radix Et Rhizoma Rhei energy activating blood and removing stasis, blood pressure lowering and blood fat reducing promote that bile secretion, gallbladder shrink, the interior bile silt of mediation biliary ductuli knot, increase the bile duct contractile function, lax Oddi's sphincter increases thereby bile is discharged, and plays the promoting the function of the gallbladder to alleviate jaundice effect, thereby Radix Et Rhizoma Rhei has promoting the function of the gallbladder to alleviate jaundice, protects the liver exsiccation.
The present invention's Chinese medicine composition for animals is first with the Radix Pulsatillae, the Radix Astragali, the Rhizoma Atractylodis Macrocephalae, Radix Isatidis compatibility, effect with strengthening vital QI to eliminate pathogenic factors, heat-clearing and toxic substances removing, enhancing body non-specific immunity, can therapy apparatus body immunity function low, immunologic tolerance, endogenous toxin and stress wait due to multiple atypical disease, growth performance descends, and humid diseases such as anemopyretic cold, influenza, bursal disease, intestinal poison syndrome.In chicken group's clinical trial, the present invention's Chinese medicine composition for animals can effectively improve chicken group's feed intake and body weight, reduce material anharmonic ratio and mortality rate, increase unit volume leukocyte and erythrocytic quantity, improve chicken group's antibody horizontal, illustrate that the present invention's Chinese medicine composition for animals can effectively improve the birds growth performance, strengthen birds body non-specific immunity, and evident in efficacy, be worth in market, promoting the use.
The specific embodiment
Below the invention will be further described for the description by the specific embodiment, but this is not to be limitation of the present invention, those skilled in the art are according to basic thought of the present invention, can make various modifications or improvement, but only otherwise break away from basic thought of the present invention, all within the scope of the present invention.
Embodiment 1 preparation veterinary medicine pre-mixing agent of the present invention
1) the weighting raw materials 25g Radix Pulsatillae, the 25g Radix Astragali, the 10g Rhizoma Atractylodis Macrocephalae, 10g Radix Isatidis, 10g Radix Scutellariae, 10g Radix Bupleuri, 10g Radix Et Rhizoma Rhei;
2) Radix Pulsatillae, the Radix Astragali, Radix Isatidis, Radix Scutellariae, Radix Et Rhizoma Rhei crushed after being dried are become coarse powder, decoct with water and extract 3 times: the decocting of 8 times of amounts of the 1st adding medical material boiled the leaching decocting liquid 3 hours; The decocting that adds 5 times of amounts of medical material for the 2nd time boiled the leaching decocting liquid 2 hours; The decocting that adds 3 times of amounts of medical material for the 3rd time boiled 1 hour, difference leaching decocting liquid, and it is standby to merge 3 decocting liquids;
3) Rhizoma Atractylodis Macrocephalae, Radix Bupleuri are ground into coarse powder, add 8 times of amount distilled water immersions 45 minutes, heating decocted 3 hours then, collect volatile oil and store for future use in addition, and the leaching decocting liquid is standby;
4) the residue medicinal residues after filtering in the step 3) are decocted with water 2 times: the decocting of 5 times of amounts of the 1st adding medical material boiled 2 hours, and the decocting of 3 times of amounts of the 2nd adding medical material boiled 1 hour; The leaching decocting liquid is standby with the decocting liquid merging of gained in this twice decocting liquid and the step 3) respectively;
5) with step 2) and step 4) in the decocting liquid of gained merge, amalgamation liquid is evaporated to 200ml; Then will be in the concentrated solution add 300ml ethanol, and to regulate pH value with 10%NaOH solution be 7.0, standing over night is got supernatant;
6) with behind the supernatant decompression recycling ethanol of gained in the step 5) with step 3) in the volatile oil of gained merge, it is standby that spray drying gets the 10.8g extract powder;
7) extract powder that step 6) is made, by having the veterinary medicament preparation method now (referring to " veterinary drug preparation production application handbook (Jiangxi science tech publishing house, in December, 1994 publication), " pharmaceutic adjuvant complete works " (Sichuan science tech publishing house, publication in January nineteen ninety-five), two (Chinese agriculture publishing houses of " Chinese veterinary drug allusion quotation " version in 2005, publish in April, 2006)), make 100g veterinary medicine pre-mixing agent of the present invention.
Embodiment 2: prepare veterinary medicine oral administration solution of the present invention
1) the weighting raw materials 22.5g Radix Pulsatillae, the 22.5g Radix Astragali, the 12.5g Rhizoma Atractylodis Macrocephalae, 12.5g Radix Isatidis, 12.5g Radix Scutellariae, 12.5g Radix Bupleuri, 5g Radix Et Rhizoma Rhei.
2) Radix Pulsatillae, the Radix Astragali, Radix Isatidis, Radix Scutellariae, Radix Et Rhizoma Rhei crushed after being dried are become coarse powder, decoct with water and extract 3 times: the decocting of 8 times of amounts of the 1st adding medical material boiled the leaching decocting liquid 3 hours; The decocting that adds 5 times of amounts of medical material for the 2nd time boiled the leaching decocting liquid 2 hours; The decocting that adds 3 times of amounts of medical material for the 3rd time boiled 1 hour, difference leaching decocting liquid, and it is standby to merge 3 decocting liquids;
3) Rhizoma Atractylodis Macrocephalae, Radix Bupleuri are ground into coarse powder, add 8 times of amount distilled water immersions 45 minutes, heating decocted 3 hours then, collect volatile oil and store for future use in addition, and the leaching decocting liquid is standby;
4) the residue medicinal residues after filtering in the step 3) are decocted with water 2 times: the decocting of 5 times of amounts of the 1st adding medical material boiled 2 hours, and the decocting of 3 times of amounts of the 2nd adding medical material boiled 1 hour; The leaching decocting liquid is standby with the decocting liquid merging of gained in this twice decocting liquid and the step 3) respectively;
5) with step 2) and step 4) in the decocting liquid of gained merge, amalgamation liquid is evaporated to 200ml; Then will be in the concentrated solution add 300ml ethanol, and to regulate pH value with 10%NaOH solution be 6.5, standing over night is got supernatant;
6) with behind the supernatant decompression recycling ethanol of gained in the step 5) with step 3) in the volatile oil of gained merge, it is standby that spray drying gets the 10.2g extract powder;
7) extract powder that step 6) is made, by having the veterinary medicament preparation method now (referring to " veterinary drug preparation production application handbook (Jiangxi science tech publishing house, in December, 1994 publication), " pharmaceutic adjuvant complete works " (Sichuan science tech publishing house, publication in January nineteen ninety-five), two (Chinese agriculture publishing houses of " Chinese veterinary drug allusion quotation " version in 2005, publish in April, 2006)), make 100ml veterinary medicine oral administration solution of the present invention.
Embodiment 3: prepare veterinary medicine granule of the present invention
1) the weighting raw materials 24.5g Radix Pulsatillae, the 22.5g Radix Astragali, the 12g Rhizoma Atractylodis Macrocephalae, 12g Radix Isatidis, 12g Radix Scutellariae, 10g Radix Bupleuri, 7g Radix Et Rhizoma Rhei.
2) Radix Pulsatillae, the Radix Astragali, Radix Isatidis, Radix Scutellariae, Radix Et Rhizoma Rhei crushed after being dried are become coarse powder, decoct with water and extract 3 times: the decocting of 8 times of amounts of the 1st adding medical material boiled the leaching decocting liquid 3 hours; The decocting that adds 5 times of amounts of medical material for the 2nd time boiled the leaching decocting liquid 2 hours; The decocting that adds 3 times of amounts of medical material for the 3rd time boiled 1 hour, difference leaching decocting liquid, and it is standby to merge 3 decocting liquids;
3) Rhizoma Atractylodis Macrocephalae, Radix Bupleuri are ground into coarse powder, add 8 times of amount distilled water immersions 45 minutes, heating decocted 3 hours then, collect volatile oil and store for future use in addition, and the leaching decocting liquid is standby;
4) the residue medicinal residues after filtering in the step 3) are decocted with water 2 times: the decocting of 5 times of amounts of the 1st adding medical material boiled 2 hours, and the decocting of 3 times of amounts of the 2nd adding medical material boiled 1 hour; The leaching decocting liquid is standby with the decocting liquid merging of gained in this twice decocting liquid and the step 3) respectively;
5) with step 2) and step 4) in the decocting liquid of gained merge, amalgamation liquid is evaporated to 200ml; Then will be in the concentrated solution add 300ml ethanol, and to regulate pH value with 10%NaOH solution be 7.5, standing over night is got supernatant;
6) with behind the supernatant decompression recycling ethanol of gained in the step 5) with step 3) in the volatile oil of gained merge, it is standby that spray drying gets the 10.5g extract powder;
7) extract powder that step 6) is made, by having the veterinary medicament preparation method now (referring to " veterinary drug preparation production application handbook (Jiangxi science tech publishing house, in December, 1994 publication), " pharmaceutic adjuvant complete works " (Sichuan science tech publishing house, publication in January nineteen ninety-five), two (Chinese agriculture publishing houses of " Chinese veterinary drug allusion quotation " version in 2005, publish in April, 2006)), make 100g veterinary medicine granule of the present invention.
More than the quality control result of three embodiment as shown in table 1:
The veterinary medicine quality control result of table 1 embodiment of the invention 1-3:
| Embodiment 1 | Embodiment 2 | Embodiment 3 | |
| Baicalin (qualitative) | Positive | Positive | Positive |
| Baicalin (qualitative) | Positive | Positive | Positive |
| Radix Pulsatillae extract (qualitative) | Positive | Positive | Positive |
| Radix Et Rhizoma Rhei extract (qualitative) | Positive | Positive | Positive |
| Astragalus polysaccharides (qualitative) | Positive | Positive | Positive |
| Anemoside B4 content (g/g) | ?0.5％ | ?0.4％ | ?0.45％ |
Below by concrete test example beneficial effect of the present invention is described further.
Test example 1 the present invention Chinese medicine composition for animals is to the influence of chicken growth performance
＜test method 〉
1. the veterinary medicine made of test drug: embodiment of the invention 1-3.
2. experimental animal: 20 ages in days " blue or green foot fiber crops " A1 is 10300 of broiler, and body condition is good, does not have the morbidity sign, is provided by Sichuan nation standing grain agricultural science and technology company limited subordinate chicken house.
3. animal grouping and feeding and management:
To test with chicken and be divided into 4 groups at random, 3 parallel test groups, 1 matched group.Mode that adopt to concentrate spice is respectively at 2 growth stage (21-28 ages in days of test group chicken group, the 51-58 age in days) by the concentration that every day, material was measured 5 ‰ (W/W) veterinary medicine of the present invention of feeding, allow chicken group free choice feeding in 12h finish, the residue 12h basal diet of feeding, the whole process of the matched group basal diet of only feeding.
In the whole test stage, the chicken group is inoculated newcastle, bird flu (H5N1 hypotype, Re-5 strain according to following immune programme for children simultaneously; The H9 hypotype) vaccine:
16 ages in days:
The H 5 N 1 avian influenza hypotype, Re-5 strain: 0.5ml/ is (intramuscular injection) only, Liaoning Yikang Biological Products Co., Ltd., lot number: 20100203;
30 ages in days:
Newcastle disease, bird flu (H9 hypotype) bivalent inactivated vaccine (La Sota strain+WD strain): 0.5ml/ be (intramuscular injection) only, the sharp biologics company limited in sea, Shanghai, lot number: 20100310.
4. growth performance is measured:
After raising beginning and finishing, calculate gain in weight, feed intake, material anharmonic ratio, mortality rate, wherein:
Gain in weight: respectively at the 21d of test, 28d, 51d, by only weighing on an empty stomach, calculates the body weight change of chicken group at 2 growth stages (21-28 age in days, 51-58 age in days) at 58d morning;
Feed intake: respectively at the 21d of test, 28d, 51d, 58d morning, feed consumption is respectively organized in weighing;
Material anharmonic ratio: gain in weight (kg) in food consumption total amount (kg)/same feeding period in material anharmonic ratio=feeding period.
5. date processing and analysis:
Data are carried out one factor analysis of variance with spss13.0 software, and group difference adopts the Duncan multiple comparisons, and significant level is 0.05.Data result mean+SD (means ± S.D) expression.
＜result of the test 〉
The result of the test of the veterinary medicine pre-mixing agent of the embodiment of the invention 1:
The gain in weight of chicken group after 2 growth stages (21-28 age in days, 51-58 age in days) raise to finish, feed intake, material anharmonic ratio, mortality rate the results are shown in Table 2.
The veterinary medicine pre-mixing agent of table 2 embodiment of the invention 1 is to the influence of chicken gain in weight, feed intake, material anharmonic ratio, mortality rate
Annotate: colleague's shoulder marking-up parent phase is with representing zero difference significantly (P＞0.05); The female different table of colleague's shoulder marking-up is shown with significant difference (P＜0.05).
As can be seen from Table 2, when the chicken group is in the 21-28 age in days, every chicken average weight gain of test group 0.160kg, every chicken average weight gain of matched group 0.168kg, the gain in weight difference of 2 groups is remarkable (P＞0.05) not; When the chicken group is in the 51-58 age in days, every chicken average weight gain of test group 0.356kg, every chicken average weight gain of matched group 0.325kg, the gain in weight significant difference of 2 groups (P＜0.05).
In the growth stage of 21-28 age in days, test group chicken group's feed intake recruitment is bigger than matched group; In the growth stage of 51-58 age in days, the feed intake of test group is greater than matched group, but the recruitment of searching for food is not as good as matched group.Simultaneously by table 2 as seen, the chicken group significantly is lower than matched group in the material anharmonic ratio of 2 growth stages, and mortality rate significantly is lower than matched group.
The veterinary medicine oral administration solution of the embodiment of the invention 2 and the veterinary medicine granule of the embodiment of the invention 3 have the similar test result:
In test to the veterinary medicine oral administration solution of the embodiment of the invention 2, when the chicken group is in the 21-28 age in days, every chicken average weight gain of test group 0.19kg, every chicken average weight gain of matched group 0.16kg, the gain in weight significant difference of 2 groups (P＜0.05); When the chicken group is in the 51-58 age in days, every chicken average weight gain of test group 0.389kg, every chicken average weight gain of matched group 0.345kg, the gain in weight significant difference of 2 groups (P＜0.05).Test group chicken group's the recruitment of searching for food, all increases than matched group during the 51-58 age in days to some extent at the 21-28 age in days, and simultaneously, the chicken group significantly is lower than matched group in the material anharmonic ratio of 2 growth stages, and mortality rate significantly is lower than matched group;
In test to the veterinary medicine granule of the embodiment of the invention 3, when the chicken group is in the 21-28 age in days, every chicken average weight gain of test group 0.175kg, every chicken average weight gain of matched group 0.151kg, the gain in weight significant difference of 2 groups (P＜0.05); When the chicken group is in the 51-58 age in days, every chicken average weight gain of test group 0.376kg, every chicken average weight gain of matched group 0.342kg, the gain in weight significant difference of 2 groups (P＜0.05).Test group chicken group's the recruitment of searching for food, all increases than matched group during the 51-58 age in days to some extent at the 21-28 age in days, and simultaneously, the chicken group significantly is lower than matched group in the material anharmonic ratio of 2 growth stages, and mortality rate significantly is lower than matched group.
At present, the research institution of lot of domestic and foreign turns to from pure natural plant effective component extracting to regulate and control environment in the livestock and poultry alimentary tract, and its little ecology is regulated and control, to reduce the use of antibiotic as the poultry growth promoter.This test is the discovering of growth performance of broiler to 10300 20 ages in days " blue or green foot fiber crops " A1 under normal cultivating condition, 2 growth stage (21-28 ages in days the chicken group, the 51-58 age in days) use veterinary medicine of the present invention after 2 weeks respectively, can obviously improve chicken group's feed intake, reduce material anharmonic ratio and mortality rate, illustrate that thus the present invention's Chinese medicine composition for animals can effectively improve the price of deed, reduce mortality rate, strengthen the immunity of animal body.
Test example 2 the present invention Chinese medicine composition for animals is to the influence of chicken peripheral blood leucocyte, erythrocyte number
＜test method 〉
1. test drug: with test example 1.
2. experimental animal: with test example 1.
3. animal grouping and feeding and management: with test example 1.
4. the mensuration of chicken peripheral blood leucocyte and RBC number:
With the chicken group of test group, matched group at 60 ages in days respectively with 16 parts of wing venous blood collections.With conventional method (" zoophysiology experiment " (the 2nd edition), Yang Xiuping chief editor, Higher Education Publishing House, in August, 2009 publication) counting leukocyte and erythrocyte.
5. date processing and analysis: with test example 1.
＜result of the test 〉
The result of the test of the veterinary medicine pre-mixing agent of the embodiment of the invention 1:
With the chicken group of test group, matched group after 60 ages in days are respectively with 16 parts of wing venous blood collections, with conventional method (" zoophysiology experiment " (the 2nd edition), Yang Xiuping chief editor, Higher Education Publishing House, publish in August, 2009) counting leukocyte and erythrocyte, concrete outcome sees Table 3.
The feed variation (n=16) of chicken leukocyte behind the veterinary medicine pre-mixing agent of the embodiment of the invention 1, erythrocyte number of table 3
Annotate:
*The unit of expression leukocyte, erythrocyte number is respectively: * 10
9/ L; * 10
12/ L.
By result of the test table 3 as can be known, the average leukocyte count of test group is 2.73, and the average leukocyte count of matched group is 2.40, the result after variance analysis, significant difference (P＜0.05); And the mean corpuscular number of test group is 2.93, and matched group mean corpuscular number is 2.57, data by variance analysis after, significant difference (P＜0.05).The mean corpuscular number and the average leukocyte count that are test group all are significantly higher than matched group.
The veterinary medicine oral administration solution of the embodiment of the invention 2 and the veterinary medicine granule of the embodiment of the invention 3 have the similar test result:
In the test to the veterinary medicine oral administration solution of the embodiment of the invention 2, the average leukocyte count of test group is 2.88, and the average leukocyte count of matched group is 2.47, the result after variance analysis, significant difference (P＜0.05); And the mean corpuscular number of test group is 3.10, and matched group mean corpuscular number is 2.69, data by variance analysis after, significant difference (P＜0.05);
In the test to the veterinary medicine granule of the embodiment of the invention 3, the average leukocyte count of test group is 2.89, and the average leukocyte count of matched group is 2.54, the result after variance analysis, significant difference (P＜0.05); And the mean corpuscular number of test group is 2.97, and matched group mean corpuscular number is 2.51, data by variance analysis after, significant difference (P＜0.05).
In recent years, existing research report is pointed out, contains biological active substances in the Chinese herbal medicine, can play the stimulation growth of animals or poultry, keeps animal internal milieu normal equilibrium.Fundamentally protect, keep the holistic health of poultry, enhancing body's immunological function, beneficial microbe group in the control agent gives full play to and improves the prophylactic potential ability of body itself.The power of immunity of organisms is relevant with the quantity of immunocyte, thereby the quantity of immunocyte can reflect the immune level of body to a certain extent.Leukocyte is an important component part of body defending system, can engulf foreign body and produce antibody, cures, resists the cause of disease invasion at body injury and the immunology of disease is played an important role.The existing erythrocyte that studies have shown that participates in the many important physical functions of body, its immunologic function is that other immunocyte institutes are irreplaceable, it and leukocyte are being brought into play important effect aspect the body non-specific immunity together, thereby this test is by counting chicken peripheral blood leucocyte, erythrocytic quantity, thereby can understand the influence of the present invention's Chinese medicine composition for animals to the chicken non-specific immunity to a certain extent.
As seen from the experiment, the mean corpuscular number of three test group and average leukocyte count are significantly higher than matched group respectively.Explanation thus, chicken can be improved unit volume leukocyte, erythrocytic quantity after taking veterinary medicine of the present invention, thereby the non-specific immunity that improves the chicken group is had positive role.
Test example 3 the present invention Chinese medicine composition for animals is to the influence of chicken serum antibody
＜test method 〉
1. test drug: with test example 1.
2. experimental animal: with test example 1.
3. animal grouping and feeding and management: with test example 1.
4. the chicken serum antibody horizontal detects:
In chicken group 60 ages in days, respectively with the wing venous blood collection, do the chicken serum antibody test at test group and matched group.Detecting index is newcastle disease antibody horizontal, avian influenza antibody horizontal, all adopts indirect hemagglutination test.
5. date processing and analysis: with test example 1.
＜result of the test 〉
The result of the test of the veterinary medicine pre-mixing agent of the embodiment of the invention 1:
The employing indirect hemagglutination test with the wing venous blood collection, has detected the antibody horizontal of test group, matched group chicken group's newcastle and bird flu in chicken group 60 ages in days, the results are shown in Table 4.
Table 4 chicken serum antibody test result
Annotate: n represents sample number; ND represents newcastle; AI represents bird flu; AI (H5Re-5), AI (H9) represent 2 hypotypes of bird flu respectively;
Expression average antibody titre.
As shown in Table 4, test group newcastle average antibody titre is 10.5 to be higher than matched group 8.4, significant difference (P＜0.05); And when detecting bird flu average antibody titre, bird flu H5Re-5 average antibody titre is 6.1 to be higher than matched group 5.5, significant difference (P＜0.05); Bird flu H9 average antibody titre is 7.5 to be higher than matched group 6.3, significant difference (P＜0.05).
The veterinary medicine oral administration solution of the embodiment of the invention 2 and the veterinary medicine granule of the embodiment of the invention 3 have the similar test result:
In the test to the veterinary medicine oral administration solution of the embodiment of the invention 2, test group newcastle average antibody titre is 11.7 to be higher than matched group 9.4, significant difference (P＜0.05), and when detecting bird flu average antibody titre, also obtained similar result.The average antibody titre of each bird flu hypotype of test group all is significantly higher than matched group;
In the test to the veterinary medicine granule of the embodiment of the invention 3, test group newcastle average antibody titre is 11.5 to be higher than matched group 8.2, significant difference (P＜0.05), and when detecting bird flu average antibody titre, also obtained similar result.The average antibody titre of each bird flu hypotype of test group all is significantly higher than matched group.
This research has been estimated the present invention's Chinese medicine composition for animals to strengthening the influence of newcastle, bird flu (H5Re-5 hypotype, H9 hypotype) immune effect of vaccine by detecting chicken group serum antibody titer.By The above results as can be known, in normal immunity inoculation, adopt veterinary medicine of the present invention to be used, can play synergism, can effectively increase immune effect,, more can improve the antibody horizontal of chicken compared with simple dependence vaccine.
Claims (9)
1. preparation method that is used for the enhancing body non-specific immunity, improves the veterinary medicine of growth performance, this method comprises:
1) the weighting raw materials Radix Pulsatillae, the Radix Astragali, the Rhizoma Atractylodis Macrocephalae, Radix Isatidis, Radix Scutellariae, Radix Bupleuri, Radix Et Rhizoma Rhei;
2) Radix Pulsatillae, the Radix Astragali, Radix Isatidis, Radix Scutellariae, Radix Et Rhizoma Rhei are pulverized, add the decocting that 3-8 doubly measures and boiled 1-4.5 hour, the leaching decocting liquid is standby;
3) Rhizoma Atractylodis Macrocephalae, Radix Bupleuri are pulverized, added the decocting that 3-8 doubly measures and boiled 1-3 hour, it is standby to collect volatile oil, and the leaching decocting liquid is standby;
4) with filter in the step 3) medicinal residues add the decocting that 3-8 doubly measures and boiled 1-3 hour, the leaching decocting liquid merges standby with the decocting liquid of gained in the step 3);
5) with step 2) in gained decocting liquid and the step 4) collecting decoction of gained merge, amalgamation liquid is concentrated, make every 1ml concentrated solution be equivalent to contain the 0.5g crude drug; Add ethanol then in concentrated solution, regulating pH value is 5.0～7.5, leaves standstill, and it is standby to get supernatant;
6) with behind the supernatant decompression recycling ethanol of gained in the step 5) with step 3) in the volatile oil of gained merge, drying is made extract powder;
7) extract powder that step 6) is made adds the pharmacy acceptable auxiliary and makes described veterinary medicine, makes every 1g or every 1ml veterinary medicine be equivalent to contain the 1g crude drug.
2. preparation method according to claim 1, the weight portion proportioning of wherein said crude drug is:
The Radix Pulsatillae 10～30 Radixs Astragali 10～30 Rhizoma Atractylodis Macrocephalaes 5～15
Radix Isatidis 5～15 Radix Scutellariaes 5～15 Radix Bupleuri 5～15
Radix Et Rhizoma Rhei 5～15.
3. preparation method according to claim 2, the weight portion proportioning of wherein said crude drug is:
The Radix Pulsatillae 20～30 Radixs Astragali 20～30 Rhizoma Atractylodis Macrocephalaes 10～15
Radix Isatidis 10～15 Radix Scutellariaes 10～15 Radix Bupleuri 10～15
Radix Et Rhizoma Rhei 5～10.
4. preparation method according to claim 3, the weight portion proportioning of wherein said crude drug is:
The Radix Pulsatillae 25 Radixs Astragali 25 Rhizoma Atractylodis Macrocephalaes 10
Radix Isatidis 10 Radix Scutellariaes 10 Radix Bupleuri 10
Radix Et Rhizoma Rhei 10.
5. preparation method according to claim 1, wherein in described step 2) in, described decoction divides to be carried out for 3 times, and the decocting of 8 times of amounts of the 1st adding medical material boiled 3 hours, the decocting that adds 5 times of amounts of medical material for the 2nd time boiled 2 hours, and the decocting of 3 times of amounts of the 3rd adding medical material boiled 1 hour.
6. preparation method according to claim 1 wherein in described step 3), further comprises the step that crude drug is soaked after adding water, before decocting, soak time is 45 minutes.
7. preparation method according to claim 1, wherein in described step 4), described decoction divides to be carried out for 2 times, and the decocting of 5 times of amounts of the 1st adding medical material boiled 2 hours, and the decocting of 3 times of amounts of the 2nd adding medical material boiled 1 hour.
8. preparation method according to claim 1 wherein adds alcoholic acid amount and is that to make the ethanol final concentration be 60% amount in the step 5); The reagent of regulating pH value is 10%NaOH solution.
9. according to each described preparation method among the claim 1-8, wherein said veterinary medicine is oral solution, granule, soluble powder or pre-mixing agent.
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