CN103494914B - Compound radix astragali immune activation injection - Google Patents
Compound radix astragali immune activation injection Download PDFInfo
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Abstract
The invention relates to immune activation injection and a preparation method. Used as a diluent, the injection can enhance the immune effects of vaccines, especially serums and strongly increase the antibacterial efficacy of antibiotic powder injection. Independently injected, the injection can protect livers and benefit gallbladders, reduce fever and improve the growth performances of animal bodies. The preparation method is characterized by comprising the following steps: cutting 10-20g of radix astragali and 5-10g of dried tangerine peel into slices and concentrating the slices with ethanol after decocting the slices, thus preparing a solution A for standby use; cutting off 5-10g of radix bupleuri, adding water to distil radix bupleuri and adding PEG400 to dissolve radix bupleuri, thus preparing a solution B for standby use; adding the solution B to the solution A, adding water for injection until the volume is 85-95ml, using a pH regulator, adding 0.1-0.2g of activated carbon, carrying out micro-boiling for one hour, cooling the solution, adding 1ml of benzyl alcohol and then achieving a constant volume of 100ml; carrying out filtering, filling and sealing as well as sterilizing, thus obtaining the injection. The injection is low in preparation cost, has good effects whether being used independently or serving as the diluent and especially has obvious synergistic effects on ceftiofur sodium.
Description
The application is application number: 201210224030.1, and patent name is: the divisional application of the patent of invention of " preparation method of Immunostimulatory injection for livestock ", the original bill applying date: on June 30th, 2012.
Technical field
The present invention relates to a kind of preparation method of injection, especially can use as injection diluent or inject separately the preparation method of the injection used.Also compound astragalus membranaceus medicine is related to.
Background technology
In the prevention and therapy of livestock and poultry, employ various types of lyophilizing shape bacterin preparation and Powdered powder injection formulation, injection or drop after need diluting in proportion when Clinical practice.The diluent that current veterinary often uses has injection normal saline and glucose injection two kinds, and they have weak point on using function.Glucose injection can be taken into account and provide energy to animal body while dilution medicine, but its pH value is about 3.0, far below the best acid-base value (pH=7-8) of diluting vaccine and most antibiotics powder pin, the pH value of injection normal saline is slightly high is 4-5, does not also reach best use value and cannot provide energy.To such an extent as to Clinical practice link, there is veterinarian independently to add sodium bicarbonate to improve the pH value of medicinal liquid.Such use is inconvenient.
In addition, injection normal saline and glucose injection due to composition simple, production firm is numerous, there are problems such as carrying disease germs, pH value change, holding time are short qualitatively.
Chinese herbal medicine is the material that the mankind apply the earliest, the effect of preventing and curing diseases can be played, and toxic and side effects is little, have no drug resistance, hazard residue can not be produced in edible animal products, also can avoid that the pathogenic microorganism using chemicals to cause in a large number develops immunity to drugs, the problem such as livestock products drug residue and environmental pollution.Take as the leading factor with the traditional theory of relation between the physical property of natural Chinese medicinal herb, thing taste, thing, be aided with the injection that modern Chinese medicine effective ingredient extractive technique and drug preparation technique make and be referred to as Chinese medicine injection, this series products is subject to concern and the high praise of relevant expert scholar day by day because of advantages such as its diverse in function, pure natural, environmental protections.
Summary of the invention
Main purpose of the present invention aims to provide a kind of pure natural multiduty Chinese medicine immune activation injection and preparation method, prepared injection is made diluent and is used the immune effect that can strengthen vaccine particularly serum, the antibacterial efficacy of powerful antibiotic potentiator injectable powder; Independent injection can hepatic cholagogic, bring down a fever, improve animal body growth performance.
For achieving the above object, the technical solution adopted in the present invention is: a kind of preparation method of Immunostimulatory injection for livestock, and step comprises:
A, Radix Astragali 10-20g, Pericarpium Citri Reticulatae 5-10g, section, adds 5 times of weight water for injection soak by water 2 times, 2 hours first times, second time 1 hour, collecting decoction, filter, filtrate is concentrated into 2 times of Chinese crude drug weight water yields, slowly add ethanol when being chilled to 40 DEG C, make alcohol content reach 75%, fully stir, leave standstill 12 hours, leaching supernatant, residue adds 75% appropriate amount of ethanol, stir evenly, leave standstill 12 hours, filter, merge ethanol, reclaim ethanol to without alcohol taste, obtain A liquid for subsequent use;
B, Radix Bupleuri 5-10g, cut off, add water warm macerating, through 4 times of weight water distillations, collects just distillate, then redistillation, and collect double distilled liquid, add the PEG400 of 20-40ml, stir and oil is dissolved completely, obtained B liquid is for subsequent use;
C, B liquid to be added in A liquid, add and inject water to 85-95ml, use pH adjusting agent to regulate pH to be 7.0-7.5, add active carbon 0.1-0.2g, micro-ly to boil 1 hour, let cool, after adding benzyl alcohol 1ml, be settled to 100ml;
D, filtration, embedding, sterilizing and get final product.During fill, can fill in cillin bottle, ampoule bottle, vial, such as 5ml, 10ml, 100ml etc. that loading amount is conventional.
Described injection is through 115 DEG C, pressure 0.2MPa sterilizing 30 minutes.
Described pH adjusting agent is the sodium hydroxide solution of 0.5mol/L or the hydrochloric acid solution of 0.5mol/L.During actual preparation, according to circumstances regulate, pH value is high to go beyond the scope, and acceptable salts acid solution regulates, and pH value is lower than this scope, and available hydrogen sodium hydroxide solution regulates.
Preferably, described Radix Astragali 15-20g, Pericarpium Citri Reticulatae 7-10g, Radix Bupleuri 7-10g.
Preferably, described PEG400 consumption is 30-40ml.
Benzyl alcohol is the preferred specific antiseptic of the present invention.PEG400 is preferred specific fat-soluble solvent, to a lot of Effective Component of Chinese Medicine as protein, aminoacid, polysaccharide, vitamin, trace element, volatile oil have good affinity, and is conducive to the pH stability of preparation.
In the present invention, Radix Astragali major function is for strengthening animal body specific immune function and non-specific immunity, sorption enhanced is the function of energy stimulating expression of macrophage and T cell after astragalus polysaccharides in animal body, make E ring form cell number to increase, Cytokines Production, promote that interleukin induces, and make animal body produce endogenous interferon, thus reach antiviral object.Identical with the Antiviral Mechanism of vaccine, the immune cooperation of body can be induced, reach synergetic immunity potentiation.Better dilution effect is embodied on serum class preparation, and the non-specific immunity of the Radix Astragali can improve the deficiency of serum antibody specific immunity on effect duration.In addition the antiinflammatory of the Radix Astragali, diuresis, blood pressure lowering, antipyretic, the multiple biological activity such as to protect the liver, there is effect of heat clearing and damp drying, eliminating fire and detoxication, be applicable to the auxiliary treatment of animal viral disease and bacterial disease.
The Radix Bupleuri selected in the present invention is because its volatile oil has effect of antiinflammatory, regulate body temperature, antipyretic-antalgic, compared with synthesizing antipyretic analgesic with aminophenazone, dipyrone etc., can not the self-resistance of damaged animal body, and bring down a fever and do not rebound.Select Pericarpium Citri Reticulatae, be because it has the function of strengthening the spleen stomach invigorating of helping digestion of regulating the flow of vital energy, animal body fluid balance can be improved, promote animal appetite, improve feed digestion utilization rate and animal product quality.
The advantage of immune activation injection of the present invention is in particular in:
1. use approach is various, can dilute the conventional preparation such as vaccine, serum, antibiotic powder pin of livestock and poultry cultivation, has the function of bringing down a fever of the health care of the growing state improving poultry and antiinflammatory, regulate body temperature during independent injection.
2, natural Chinese medicines preparation toxic and side effects is little, has no drug resistance, and can not produce hazard residue in edible animal products, also can avoid that the pathogenic microorganism using chemicals to cause in a large number develops immunity to drugs, the problem such as livestock products drug residue and environmental pollution.
3, reasonably extract and preparation process operation simply very convenient, cost control is low, be conducive to commercial production needs.
4, stable pH value range, is applicable to the antibacterial environment for use of antibiotic powder pin, can strengthens its sterilizing function, particularly remarkable, such as remarkable to the potentiation of ceftiofur sodium to beta-lactam, Tetracyclines powder pin synergistic effect.Simultaneously reduce or offset antibiotic antibacterial time produce toxic and side effects.
5, synergistic function is had to the non-specific immunity of vaccine, especially to suffering from immunosuppressive disease or immune unresponsive poultry are respond well; There is complementary action to the specific immune function defect of serum, activate animal body active immune function, extend the time of immunoprotection.
Detailed description of the invention
The Chinese crude drug Radix Astragali used in embodiment, Radix Bupleuri, Pericarpium Citri Reticulatae (medicinal decoction pieces) are purchased from prepared slices of Chinese crude drugs factory of Chongqing City, benzyl alcohol (AR) is purchased from ShouGuang Luyuan Salt Chemical Co., Ltd., PEG400(pharmaceutical grade) and medical ethanol be purchased from Gutian, Nanjing Chemical Co., Ltd., activated carbon is purchased from the Zhong Mountain, Chongqing activated carbon Manufacturing Co., Ltd.
Embodiment one
Immune activation injection (1ml is equivalent to crude drug in whole 0.4g)
1, formula
Radix Astragali 20g, Radix Bupleuri 10g, Pericarpium Citri Reticulatae 10g, pH adjusting agent 0.2g, activated carbon 0.2g, PEG400 consumption 40ml, benzyl alcohol 1ml, water for injection adds to 100ml.
2: preparation method
The first step: get the recipe quantity Radix Astragali, Pericarpium Citri Reticulatae, section, adds 5 times of weight water for injection soak by water 2 times, 2 hours first times, second time 1 hour, collecting decoction, filter, filtrate is concentrated into 2 times of Chinese crude drug weight water yields, slowly add ethanol when being chilled to 40 DEG C, make alcohol content reach 75%, fully stir, leave standstill 12 hours, leaching supernatant, residue adds 75% appropriate amount of ethanol, stir evenly, leave standstill 12 hours, filter, merge ethanol, reclaim ethanol to without alcohol taste, obtain A liquid for subsequent use.
Second step: get recipe quantity Radix Bupleuri and cut off, add water warm macerating, through 4 times of weight water distillations, collects just distillate, then redistillation, and collect double distilled liquid, add recipe quantity PEG400, stir and oil is dissolved completely, obtained B liquid is for subsequent use.
3rd step: B liquid is added in A liquid, add and inject water to 95ml, the sodium hydroxide solution regulation system pH using 0.5mol/L is 7.5, add recipe quantity active carbon, micro-ly to boil 1 hour, let cool, 100ml is settled to after adding recipe quantity benzyl alcohol, filter, embedding, through 115 DEG C, pressure 0.2MPa sterilizing 30 minutes and get final product.
Embodiment two
Immune activation injection (1ml is equivalent to crude drug in whole 0.3g)
1, formula
Radix Astragali 15g, Radix Bupleuri 7.5g, Pericarpium Citri Reticulatae 7.5g, pH adjusting agent 0.2g, activated carbon 0.15g, PEG40030ml, benzyl alcohol 1ml, water for injection adds to 100ml.
2: preparation method: identical with method described in embodiment 1.
Embodiment three
Immune activation injection (1ml is equivalent to crude drug in whole 0.2g)
1, formula
Radix Astragali 10g, Radix Bupleuri 5g, Pericarpium Citri Reticulatae 5g, pH adjusting agent 0.1g, activated carbon 0.1g, PEG40020ml, benzyl alcohol 1ml, water for injection adds to 100ml.
2: preparation method: identical with method described in embodiment 1.
One, the accelerated stability test of immune activation injection
1, materials and methods: Example 1-3 immune activation injection, hot and humid high light Acceleration study is carried out according to " Chinese veterinary pharmacopoeia " version in 2010 annex medicine stability experimental technique, by commercially available back, place 6 months in the medicine stability experimental box (Chongqing immortality experimental apparatus factory) of temperature 40 ± 2 DEG C, relative humidity 75 ± 5%, illumination 4500 ± 500LX, sample respectively once 1st, 2,3,6 the end of month at experimental session, the color of injection, pH value, content are investigated.
2, result: as shown in Table 1, immune activation injection of the present invention is in accelerated test after 6 months, and color, pH value, each component content have no significant change, and have good stability,
The Acceleration study result of table 1, Immunostimulatory injection for livestock
Two, the impact of immune activation injection on newcastle disease and infections bursal disease vaccine immunity power is tested
Preparation chicken ND I is, ND IV is, ND oil-emulsion vaccine and IBD freeze-dried vaccine to use embodiment 1 injection to dilute respectively, in 15 days, being successively inoculated in 1 group of (35) healthy nonimmune chickling by immune programme for children is experimental group 1, with method operation embodiment 2, example 3 injection, in 15 days, being successively inoculated in 2 groups of (often organizing 35) healthy nonimmune chickling by immune programme for children is experimental group 2 and 3.Use injection normal saline to dilute respectively chicken ND I is, ND IV is, ND oil-emulsion vaccine and IBD freeze-dried vaccine simultaneously, is successively inoculated in 1 group of (35) healthy nonimmune chickling as a control group by immune programme for children in 15 days.After the 4th immunity terminates 10 days, use agar gel diffusion test (AGP) to monitor ND, IBD antibody titer in serum, dynamic observation has been carried out to the change of each group of chicken spleen, bursa of Fabricius's weight simultaneously.Testing result: each embodiment group serum N D, IBD antibody titer are significantly higher than matched group (P < 0.05) is wherein the highest with the antibody titer of embodiment 1 group; In addition, embodiment 1-example 3 groups of spleens and bursa of Fabricius's weight are significantly higher than matched group (P < 0.05).Test shows, immune activation injection can be used as the immunity of the immunological adjuvant raising chicken of chicken vaccine, and embodiment 1 group is best.
Table 2, immune activation injection affect result of the test table to newcastle disease and infections bursal disease vaccine immunity power
| Implement group number | IBD tires | ND tires | Spleen (g/kg) | Thymus (g/kg) |
| Experimental group 1 | 122±2.7 | 256±5.4 | 2.21±0.11 | 0.62±0.09 |
| Experimental group 2 | 113±2.1 | 238±5.2 | 2.17±0.09 | 0.59±0.11 |
| Experimental group 3 | 108±2.2 | 230±5.0 | 2.18±0.12 | 0.57±0.08 |
| Matched group | 71±1.8 | 139±3.9 | 1.58±0.02 | 0.54±0.09 |
Three, immune activation injection is to the synergism test of antibiotic powder pin injection ceftiofur sodium
1, method and material
Choose pig farm, Yunmen town, Hechuan, veterinary clinic is diagnosed as the sick pig that pig streptococcicosis infects, be divided into A, B, C, D tri-groups at random, A group adopts embodiment 1 immune activation injection dilution injection ceftiofur sodium, B group adopts embodiment 2 immune activation injection dilution injection ceftiofur sodium, C group adopts embodiment 3 immune activation injection dilution injection ceftiofur sodium, and the contrast of D group adopts glucose injection dilution injection ceftiofur sodium.Often organize all by ceftiofur 3mg/kg intramuscular injection, injection on the one once, is used in conjunction 3.
2, result decision procedure
Observe treatment case in after 3 days, pay a return visit and record the state of an illness.
Cure: medication 3 future trouble poultry diseases disappear completely, and spirit, body temperature, appetite recover normal, and urine feces is normal.
Effective: medication 3 future trouble poultry diseases disappear substantially, and spirit, body temperature, appetite make moderate progress, and urine feces is normal.
Invalid: medication 3 future trouble poultry diseases do not disappear, and sb.'s illness took a turn for the worse or death, and urine stool colour, character are abnormal.
3, result: as can be seen from Table 3, uses the antibiotic powder pin that medicine immune activation injection of the present invention dilutes, and have significant curative effect to treatment bacterial infection of livestock, cure rate and effective percentage are all significantly higher than the matched group using glucose injection dilution.The experimental group effective percentage of product of the present invention is wherein used to reach 98%, cure about 90%; And matched group D cure rate is only 52.94%, effective percentage 81.88%.
The synergism test of table 3, injection ceftiofur sodium
Four, immune activation injection solution heat test
1, method and material
1.1 medicine
Immune activation injection, adopts preparation example 1-example 3 of the present invention.
Analgin injection, animal pharmaceutical factory of Chongqing City produces.
0.9% normal saline, laboratory is provided for oneself.
Typhoid fever attenuated vaccine, is purchased from Chinese biological technology group company.
1.2 laboratory animals: get body weight 1.5-2.0kg, for three days on end body temperature the healthy rabbits 50 of 39.0 ± 5 DEG C, be divided into 5 groups at random, often organize 10, male and female half and half.
1.3 implementation processes: front 3 groups of rabbit inject the immune activation injection of embodiment 1-example 3 respectively, and injected dose presses 0.1ml/kg.Matched group injection analgin injection, every injected dose presses 0.1ml/kg.Blank group injection 0.9% normal saline, every injected dose presses 0.1ml/kg.Each group of rabbit injects according to dosage 1ml/kg ear vein injection typhoid fever attenuated vaccine after 30 minutes, then before and after vaccinate 30,60,90,120,150,180,210, measure anus temperature in the 240min time, record data, the results are shown in Table 4.
2, result: as can be seen from Table 4, after injection typhoid fever Attenuate vaccine 30min, each group rabbit body temperature significantly rises, the 60min moment reaches peak value, the sick rabbit body temperature of immune activation injection basic controlling when 120min of embodiment 1-example 3 is normal, also the sick rabbit body temperature of basic controlling is normal when 180min for analgin injection, immune activation injection is less in each time period temperature fluctuation difference, and antipyretic response is good and be better than analgin injection.
The antipyretic result of the test of table 4, fever in rabbit (DEG C, ± S)
Claims (1)
1. an Immunostimulatory injection for livestock, is characterized in that: described Immunostimulatory injection for livestock consists of: Radix Astragali 15g, Radix Bupleuri 7.5g, Pericarpium Citri Reticulatae 7.5g, pH adjusting agent 0.2g, activated carbon 0.15g, PEG40030ml, benzyl alcohol 1ml, and water for injection adds to 100ml; The preparation method of described Immunostimulatory injection for livestock is:
The first step: get the recipe quantity Radix Astragali, Pericarpium Citri Reticulatae, section, adds 5 times of weight water for injection soak by water 2 times, 2 hours first times, second time 1 hour, collecting decoction, filter, filtrate is concentrated into 2 times of Chinese crude drug weight water yields, slowly add ethanol when being chilled to 40 DEG C, make alcohol content reach 75%, fully stir, leave standstill 12 hours, leaching supernatant, residue adds 75% appropriate amount of ethanol, stir evenly, leave standstill 12 hours, filter, merge ethanol, reclaim ethanol to without alcohol taste, obtain A liquid for subsequent use;
Second step: get recipe quantity Radix Bupleuri and cut off, add water warm macerating, through 4 times of weight water distillations, collects just distillate, then redistillation, and collect double distilled liquid, add recipe quantity PEG400, stir and oil is dissolved completely, obtained B liquid is for subsequent use;
3rd step: B liquid is added in A liquid, add and inject water to 95ml, the sodium hydroxide solution regulation system pH using 0.5mol/L is 7.5, add recipe quantity active carbon, micro-ly to boil 1 hour, let cool, 100ml is settled to after adding recipe quantity benzyl alcohol, filter, embedding, through 115 DEG C, pressure 0.2MPa sterilizing 30 minutes and get final product.
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| JP2006333703A (en) * | 2005-05-06 | 2006-12-14 | Daicho Kikaku:Kk | Animal feed additive and animal feed |
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| CN1628834A (en) * | 2004-09-10 | 2005-06-22 | 张平 | Freeze dried powder injection of Bupleurum root and its preparation process |
| CN1757305A (en) * | 2004-10-10 | 2006-04-12 | 吉林省农业科学院 | Chinese herbal feed additive and preparation method thereof |
| CN102462728A (en) * | 2010-11-17 | 2012-05-23 | 郭进军 | Medicinal composition for treating and preventing bacterial upper respiratory tract infection |
Non-Patent Citations (3)
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| 谢麟等.30.黄芪、柴胡注射液.《动物药剂的应用与制作科技全书》.化学工业出版社,2002,(第1版),第435页. * |
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