CN1660084A - Aquatic animal antibacterial medicine and preparation method and medicine feeding method thereof - Google Patents
Aquatic animal antibacterial medicine and preparation method and medicine feeding method thereof Download PDFInfo
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- CN1660084A CN1660084A CN2004101032256A CN200410103225A CN1660084A CN 1660084 A CN1660084 A CN 1660084A CN 2004101032256 A CN2004101032256 A CN 2004101032256A CN 200410103225 A CN200410103225 A CN 200410103225A CN 1660084 A CN1660084 A CN 1660084A
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- antibacterial
- aquatic animal
- oxolinic acid
- starch
- vitamin
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- 239000003814 drug Substances 0.000 title claims abstract description 31
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- 238000000034 method Methods 0.000 title claims abstract description 16
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
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- KYGZCKSPAKDVKC-UHFFFAOYSA-N Oxolinic acid Chemical compound C1=C2N(CC)C=C(C(O)=O)C(=O)C2=CC2=C1OCO2 KYGZCKSPAKDVKC-UHFFFAOYSA-N 0.000 claims abstract description 34
- 229960000321 oxolinic acid Drugs 0.000 claims abstract description 33
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Landscapes
- Farming Of Fish And Shellfish (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
An antibacterial medicine for aquatic animals contains oxolinic acid, vitamin C and starch. The antibacterial agent is prepared in a special process, and is fed to aquatic animals such as mandarin fish by a special method. Clinical tests prove that the aquatic animal antibacterial agent has low minimum inhibitory concentration and safe use, has a good prevention and treatment effect on bacterial hemorrhagic disease of mandarin fish caused by aeromonas, the prevention and treatment effective rate is about 80%, and the stability of the effective components of the antibacterial agent preparation is good.
Description
Technical field
The present invention relates to a kind of antibacterial for aquatic animal and preparation method thereof and method of feeding medicine.
Background technology
At present, all kinds of new diseases constantly occur in the aquaculture, make in the aquaculture process, need to use medicines that cultivation water environment sterilization, improvement and aquatic animal are prevented in a large number and treat, and in therapeutic process, the strong dose substrate concentration, be easy to cause aquatic animal and the residual even severe overweight of cultivation water environment Chinese medicine, a large amount of inputs of medicine also cause the change of water body physical and chemical index easily, destroy the growing environment of aquatic animal.As current, in aquaculture production, it is main drug treatment that antibiotics is adopted in the control of bacterial disease mostly, and antibiotics plays sizable effect really to the control of aquatic animal disease.But in recent years, antibiotics uses kind, dosage and frequency all to be improved largely in aquaculture, become main component in the various fisheries drug preparations as antibiotics such as medical quinolinones such as ciprofloxacin, norfloxacin (qunolones) classes, some medical novel carbostyril medicines such as ofloxacin etc. also use in breeding production in a large number in addition.But in many antibacterials, some jumping genes relevant with antibiotic resistance can be transferred in other antibacterial, thereby might cause the antibacterial relevant with human diseases also might produce drug resistance to this antibiotics.Improper use simultaneously also can cause medicine residual in aquatic products, reduces aquatic product quality, and the mankind are produced potential threat.
Summary of the invention
The object of the invention provides a kind of safe in utilization, the antibacterial for aquatic animal that the bacterial disease of aquatic animal had treatment and preventive effect and preparation method thereof and method of feeding medicine.
Technical scheme of the present invention is: a kind of antibacterial for aquatic animal, its Han You oxolinic acid, vitamin C, starch.
A kind of preparation method of antibacterial for aquatic animal, it comprises the steps:
(1), with starch in the oven dry 2 hours down of 105 ℃～110 ℃ temperature conditions, 40 mesh sieves are crossed in the cooling back;
(2), Cheng Qu oxolinic acid and vitamin C in proportion;
(3), Qu Yu the starch that oxolinic acid and ascorbic gross weight equate, make starch Yu oxolinic acid, vitamin C fully are mixed and made into female powder;
(4), female powder is fully mixed with remaining starch again.
A kind of method of feeding medicine of antibacterial for aquatic animal, it comprises the steps:
(1), in bait, adds this medicine;
(2), with bait feeding living body bait fish;
(3), the living body bait fish is thrown something and fed aquatic animal.
Oxolinic acid (Qxolinic acid (OA)) has another name called azulenyl Suo Lin acid, and oxolinic acide etc. belong to generation quinolones, is the special-purpose antibiotics of Aquatic product that use is recommended by China.The oxolinic acid bactericidal range is wide, sterilizing power is strong, escherichia coli, Salmonella, pasteurellosis bacillus, haemophilus, staphylococcus, lonely bacterium, Aeromonas, pseudomonas etc. all there is intensive antibacterial and bactericidal action, it is safe, have no side effect, oral easy absorption, tissue penetration is strong, almost completely drains from urine, does not have crossing drug resistant with all kinds of antibiotic.Oxolinic acid causes disease such as scale protrusion disease, erythroderma, gill rot, mashed tail, freshwater fish bacterial septicemia to aquatic animal because of bacterial infection, the tarda disease of eel, vibriosis, red fin, ulcer, red point, mashed tail, gill rot, the erythromelia of bull frog, ascites, enteritis, bark rot, the gill rot of shrimp, Eriocheir sinensis, black gill disease, red leg disease, fluorescence disease, carapace appendage Peptic Ulcers etc., white macula, the base plate of Trionyx sinensis Wiegmann is hemorrhage, blank, sternite redness, bacterial enteritis, ascites, mashed neck tail etc. all have good curative effect.The residual quantity of ， oxolinic acid in the flesh of fish and fish skin is less aspect residual.
The vitamin C of the main adjuvant of Zuo Wei oxolinic acid claims ascorbic acid again, belongs to water soluble vitamins.Be that Fish are kept the necessary material of physiological function.After (1965) such as Kitamura confirm by experiment that first the normal growth of Fish need be taken the photograph people's vitamin C, studies show that more vitamin C has important effect to aspects such as the breedings of Fish, growth promoter, disease-resistant and survival rate raisings.Vitamin C keep its normal growth function by Fish and vital movement essential.Vitamin C directly influences the synthetic of collagen protein as the coenzyme of proline hydroxylase in the building-up process of collagen protein, participates in the generation of intercellular substance.The permeability, stimulation coagulation function, the resistant function of increase to infecting that reduce blood capillary are arranged, and participate in function of detoxification.Be used for various acute and chronic infectious disease or other disease to build up resistance, after being ill the auxiliary treatment of convalescent period, wound healing phase.In addition, must have vitamin C to exist in the detoxifcation of the several drugs reaction in vivo, vitamin C also can promote the absorption of ferrum, is of value to hemopoietic function.Vitamin C also participates in intravital redox reaction, participates in the metabolism of folic acid, calcium etc. and synthesizing of parahormone.Aspect disease control of aquatic animal, find that after deliberation vitamin C can influence the healing rate of Fish wound, when being deficient in vitamin C in the feedstuff, before vertebra appears in Oncorhynchi, symptom such as ocular injury; In ditch Nian, also observe vitamin C and can strengthen its pair cell infection.
The present invention compared with prior art has following advantage:
Test report is as follows:
The sample clinical experiment report
| Kind of inspection | Clinical trial | Test basis | The Ministry of Agriculture's new fishery Clinical Laboratory standard |
| Main content of the test | 1, the report of bacteriostatic test.2, to the acute toxicity test of fresh water Macrobrachium nipponensis(de Haan).3, the stability of pharmaceutical preparation is measured.4, the control of Siniperca chuatsi bacterial haemorrhage disease is tested. | ||
| Main conclusions | 1, bacteriostatic test is 1.1mg/L to the minimal inhibitory concentration of the Aeromonas pathogenic bacterium CL990909-3 bacterial strain of crab.Minimal inhibitory concentration 1.8mg/L to Trionyx sinensis (Wiegmann) Aeromonas sobria pathogenic bacterium TL97424 bacterial strain.Minimum antibacterial 0.91mg/L to Aeromonas hydrophila pathogenic bacterium ATCC51208 bacterial strain.Minimal inhibitory concentration to freshwater fish bacterial septicemia pathogen Aeromonas hydrophila BSK-10 is 0.91mg/L.Minimal inhibitory concentration to freshwater fish bacterial septicemia pathogen Aeromonas hydrophila DF-30 is 2.1mg/L.Minimal inhibitory concentration to Vibrio anguillarum is 0.91mg/L.Minimal inhibitory concentration to the eel tarda is 0.91mg/L.Minimal inhibitory concentration to fish evil myxobacter is 1.82mg/L.2, to the acute toxicity test of fresh water Macrobrachium nipponensis(de Haan) according to operation instruction, the using dosage of product is 1mg/kg only, and to LD50 〉=500mg/kg of Macrobrachium nipponensis(de Haan), so its use is safe.3, in the control test mouthful filling sample high dose group to Siniperca chuatsi bacterial haemorrhage disease, the survival rate that mouth is irritated counteracting toxic substances group Siniperca chuatsi after 4 hours is 100%, and the survival rate of counteracting toxic substances deutostoma filling in 2 hours group Siniperca chuatsi is 90%; In the use group, the survival rate that mouth is irritated counteracting toxic substances group Siniperca chuatsi after 4 hours is 80%, and the survival rate of counteracting toxic substances deutostoma filling in 2 hours group Siniperca chuatsi is 80%; In the low dose group, the survival rate of mouthful filling counteracting toxic substances group Siniperca chuatsi after 4 hours is 50%, and the survival rate of counteracting toxic substances deutostoma filling in 2 hours group Siniperca chuatsi is 40%; The mortality rate that mouth is irritated feedstuff counteracting toxic substances group is 100%, and mouthful blank survival rate of organizing of filling feedstuff is 100%.4, the average recovery rate of three batches of De oxolinic acids of the stability of pharmaceutical preparation mensuration is 99.75%, show content of effective conformance with standard in the product; At 40 ± 2 ℃, relative humidity is that the average recovery rate of preserving after three months under 75 ± 5% conditions is 98.00%, reduces by 1.77%. | ||
The report of bacteriostatic test
One, test material:
The sample of white powdered;
Test strain:
The Aeromonas pathogenic bacterium CL990909-3 of crab
Trionyx sinensis (Wiegmann) Aeromonas sobria pathogenic bacterium TL97424
Aeromonas hydrophila pathogenic bacterium ATCC51208
Freshwater fish bacterial septicemia pathogen Aeromonas hydrophila BSK-10
Freshwater fish bacterial septicemia pathogen Aeromonas hydrophila DF-30
Vibrio anguillarum
The eel tarda
Fish evil myxobacter
Two, test method
Relevant bacteriological method during " water and effluent monitoring analytical method " that test method is write by Ministry of Agriculture's " new fisheries drug clinical trial technical specification " (provisional) and State Bureau of Environmental Protection etc. reaches carries out.
Three, sample is to the different aquatic animals of the 8 strains mensuration antibacterial, bacteriocidal concentration of causing a disease
Culture medium: bacteria culture media gravy peptone culture medium.
The preparation of bacterium liquid: pick the test strain of purification with inoculating loop, be inoculated on the gravy peptone culture medium, cultivate after 18 hours, bacterium liquid is diluted to 2,000,000 antibacterial/ml for 28 ℃.
Assay method: get 10 in sterilization test tube, add sterilization physiology salt 9ml (the 1st pipe), 5ml (2-10 pipe) with the every pipe in sterile working again.The sterile working draws pure oxolinic acid diluent 1ml to be measured and goes into to put the 1st pipe, draws 5ml behind the mixing and puts into the 2nd pipe, draws 5ml behind the mixing and puts into the 3rd pipe, does to draw 5ml behind the mixing and discard by pipe dilution the 9th pipe with this, and the 10th pipe does not add medicine and compares.
Every pipe adds the good bacterium liquid 0.5ml of dilution, 28 ℃ of cultivations behind the mixing.Pipette 1ml to aseptic culture medium after 24 hours, cultivate perusal after 24 hours, containing the concentration that minimum still is not the test tube contained drug that mixes erosion is minimal inhibitory concentration.Result of the test:
Sample is 1.1mg/L to the minimal inhibitory concentration of the Aeromonas pathogenic bacterium CL990909-3 bacterial strain of crab.
Sample is to the minimal inhibitory concentration 1.8mg/L of Trionyx sinensis (Wiegmann) Aeromonas sobria pathogenic bacterium TL97424 bacterial strain.
Sample is to the minimal inhibitory concentration 0.91mg/L of Aeromonas hydrophila pathogenic bacterium ATCC51208 bacterial strain.
Sample is 0.91mg/L to the minimal inhibitory concentration of freshwater fish bacterial septicemia pathogen Aeromonas hydrophila BSK-10.
Sample is 2.1mg/L to the minimal inhibitory concentration of freshwater fish bacterial septicemia pathogen Aeromonas hydrophila DF-30.
Sample is 0.91mg/L to the minimal inhibitory concentration of Vibrio anguillarum.
Sample is 0.91mg/L to the minimal inhibitory concentration of eel tarda.
Sample is 1.82mg/L to the minimal inhibitory concentration of fish evil myxobacter.
Acute toxicity test to the fresh water Macrobrachium nipponensis(de Haan)
One, test material
Sample, healthy Macrobrachium nipponensis(de Haan).
Two, test method
At area 0.5m
2, dark 0.5m net cage in raise 40 of fresh water Macrobrachium nipponensis(de Haan)s, support temporarily and carry out acute toxicity test after 7 days, test first three sky and stop feeding.Test is adopted and is mixed the bait oral medication, and the feedstuff feeding amount is 5% of a fresh water Macrobrachium nipponensis(de Haan) body weight.
Before the test, after the pulverizing of fresh water Macrobrachium nipponensis(de Haan) feedstuff, the fish flour of adding 20% and 0.5% feed adhesive are admixed an amount of sample in the fresh water Macrobrachium nipponensis(de Haan) feedstuff of having pulverized, add water and make pellet and 45 dryings again.Contain sample in the medicinal bait and be respectively 2,4,6,8 and 10g/Kg, the throwing amount of raising by 5% is calculated, and then is equivalent to per kilogram fresh water Macrobrachium nipponensis(de Haan) and takes in 100,200,300,400 and the 500mg sample every day.
During test, throw something and feed every day contain sample pharmaceutical chemistry once, fresh water Macrobrachium nipponensis(de Haan) quantity and be calculated to be motility rate in every net cage of statistics after throwing something and feeding continuously four days.Establish the blank group simultaneously.At least observed three times in per 24 hours, the record death toll is carried out 96 hours observed and recordeds.The water temperature of duration of test is 22 ± 2 ℃, and PH is 7.50.
Three, result of the test: mortality does not appear in the fresh water Macrobrachium nipponensis(de Haan) of each administration group and matched group, and the survival rate of matched group fresh water Macrobrachium nipponensis(de Haan) is 90%, and it is 95% that 100mg/Kg forms motility rate; It is 90% that 200mg/Kg forms motility rate; It is 97.5% that 300mg/Kg forms motility rate; It is 92.5% that 400mg/Kg forms motility rate; It is 90% that 500mg/Kg forms motility rate.Analyze through the SPSS statistical software, test group and matched group survival rate do not have significant difference, so the result shows: sample is to 96 hours LD50 〉=500mg/kg body weight of fresh water Macrobrachium nipponensis(de Haan).The results are shown in Table
Table is to the acute toxicity test of Macrobrachium nipponensis(de Haan)
| Drug level (mg/Kg) | Tried shrimp number (only) | 96h is tried shrimp survival (only) | Survival rate (%) |
| ????100 | ????40 | ????38 | ????95 |
| ????200 | ????40 | ????36 | ????90 |
| ????300 | ????40 | ????39 | ????97.5 |
| ????400 | ????40 | ????37 | ????92.5 |
| ????500 | ????40 | ????36 | ????90 |
| Matched group | ????40 | ????36 | ????90 |
Four, main conclusions
According to operation instruction, the using dosage of sample is 1mg/kg only, and to LD50 〉=500mg/kg of Macrobrachium nipponensis(de Haan), so the use of this product is safe.
Control test to Siniperca chuatsi bacterial haemorrhage disease
One, materials and methods
1, test material
1.1 experimental animal:
Test is 87.4 grams-112.9 grams with the Siniperca chuatsi body weight, available from the market of farm produce.After earlier in water temperature is 25 ℃～28 ℃ indoor cement pit, supporting a week temporarily before the test, and randomly draw five Siniperca chuatsis, blood is carried out antibacterial with muscle, hepatopancrease, heart separate, confirm to test behind the no bacterial infection.
1.2 test strain:
The pathogen Aeromonas sobria GY20010506 bacterial strain of Siniperca chuatsi bacterial haemorrhage disease (being separated by China Aquatic Science Research Institute fresh water fishery center) is inoculated in the common meat peptone culture fluid, and the 25-27 ℃ of bacteria suspension of cultivating 20-24 hour is standby.When carrying out the test of fish body, survey the half lethal dose (LD50) of bacterial strain earlier to the river Siniperca chuatsi, inoculate the fish body with the lumbar injection of doubly measuring of LD50 again, the dosage of injection is 2,000 ten thousand/only.
1.3 sample.
2, test method
2.1 test grouping
Additive capacity is 2% sample in the feedstuff, and with the 5% daily ration, feeding quantity forage fiss of throwing something and feeding, the forage fiss of the reuse 7-8% Siniperca chuatsi of throwing something and feeding.The amount that is forage fiss absorption sample is 0.100g/Kg
Test divides three groups, high dose group, and it is 1.000g/Kg that the bait lymphogranuloma inguinale is irritated dosage, and forage fiss is decaptitated after homogenate makes meat paste, irritates Siniperca chuatsi with 80g/Kg minced fish mouth again.The using dosage group, it is 0.100g/Kg that the bait lymphogranuloma inguinale is irritated dosage, irritates Siniperca chuatsi with 80g/Kg minced fish mouth again.Low dose group, mouthful filling dosage is 0.010g/Kg, irritates Siniperca chuatsi with 80g/Kg minced fish mouth again.Simultaneously every group become again respectively the Siniperca chuatsi mouth irritate the minced fish administration after 4 hours counteracting toxic substances and 2 hours deutostomas of counteracting toxic substances irritate two groups of minced fish, to estimate prevention and the therapeutic effect of this product to Siniperca chuatsi bacterial haemorrhage disease.Counteracting toxic substances dosage is 2 * 107CFU/ tail.Establish a mouthful filling normal saline counteracting toxic substances group and a mouthful filling normal saline blank group simultaneously.
2.2 test method
At the trial, take by weighing the samples of 0.500 gram, 5.000 grams and 50.00 grams respectively, after adding water and making pasty state, standardize solution is in 250 milliliters, and mouthful irritating dosage is that per kilogram Carassius auratus mouth is irritated 20ml.The amount that makes Carassius auratus take in sample is respectively 0.005g/Kg, 0.050g/Kg, 0.500g/Kg.Mouthful irritate after 60 minutes, Carassius auratus caught out, remove scale, head and fin ray, use tissue refiner, with 8000rpm homogenate 5min, make meat paste after, 4 ℃ of preservations.
In 2M * 3M net cage, raise 10 of 141.7～275.3 Siniperca chuatsis that restrain at random, begin test after the foster week temporarily.The minced fish of homogenate is sucked the 10ml syringe, and syringe connects the hard sebific duct.Siniperca chuatsi is fished for out from the pond, after weighing, insert the about 2cm of sebific duct from the mouth device, according to the body weight of Siniperca chuatsi, pour into high, normal, basic three groups of minced fish that contain sample respectively, the amount that makes Siniperca chuatsi take in minced fish is 80g/Kg.Mouth is irritated once every other day, and the counteracting toxic substances group and the blank group of irritating blank minced fish 80g/Kg by mouth are established in mouthful filling 4 days simultaneously.At least observed three times in per 24 hours, the record death toll was carried out observed and recorded 7 days.The duration of test water temperature is 25 ± 1 ℃, and PH is 7.4.
Two, result of the test
The result shows: in mouthful filling sample high dose group, the survival rate of mouthful filling counteracting toxic substances group Siniperca chuatsi after 4 hours is 100%, and the survival rate of counteracting toxic substances deutostoma filling in 2 hours group Siniperca chuatsi is 90%; In the use group, the survival rate of mouthful filling counteracting toxic substances group Siniperca chuatsi after 4 hours is 80%, and the survival rate of counteracting toxic substances deutostoma filling in 2 hours group Siniperca chuatsi is 80%; In the low dose group, the survival rate of mouthful filling counteracting toxic substances group Siniperca chuatsi after 4 hours is 50%, and the survival rate of counteracting toxic substances deutostoma filling in 2 hours group Siniperca chuatsi is 40%; The mortality rate that mouth is irritated feedstuff counteracting toxic substances group is 100%, and mouthful blank survival rate of organizing of filling feedstuff is 100%.The results are shown in following table
Table is to the prevention effect of Siniperca chuatsi bacterial haemorrhage disease
| High dose group | Use the agent group | Low dose group | Mouth is irritated the feedstuff group | The blank group | |||||
| Mouth is irritated counteracting toxic substances after 4 hours | 2 hours deutostomas of counteracting toxic substances are irritated | Mouth is irritated counteracting toxic substances after 4 hours | 2 hours deutostomas of counteracting toxic substances are irritated | Mouth is irritated counteracting toxic substances after 4 hours | 2 hours deutostomas of counteracting toxic substances are irritated | ????0 | ????0 | ||
| Mouth is irritated dosage (g/Kg) | ??0.5 | ??0.5 | ??0.050 | ??0.05 | ??0.005 | ??0.005 | ????0 | ????0 | |
| Injection Ah microbial inoculum amount | ??4000 | ??4000 | ??4000 | ??4000 | ??4000 | ??4000 | ????4000 | ????0 | |
| Test fish number | ??10 | ??10 | ??10 | ??10 | ??10 | ??10 | ????10 | ????10 | |
| Death time | First day | ??0 | ??0 | ??1 | ??1 | ??1 | ??2 | ????2 | ????0 |
| Second day | ??0 | ??1 | ??0 | ??1 | ??1 | ??1 | ????4 | ????0 | |
| The 3rd day | ??0 | ??0 | ??1 | ??0 | ??2 | ??0 | ????1 | ????0 | |
| The 4th day | ??0 | ??0 | ??0 | ??0 | ??1 | ??1 | ????1 | ????0 | |
| The 5th day | ??0 | ??0 | ??0 | ??0 | ??0 | ??0 | ????0 | ????0 | |
| The 6th day | ??0 | ??0 | ??0 | ??0 | ??0 | ??0 | ????0 | ????0 | |
| The 7th day | ??0 | ??0 | ??0 | ??0 | ??0 | ??0 | ????0 | ????0 | |
| Survival rate (%) | ??100 | ??90 | ??80 | ??80 | ??50 | ??40 | ????0 | ????100 | |
The present invention has the better prevention effect to the Siniperca chuatsi bacterial haemorrhage disease that is caused by Aeromonas, and the effective percentage of control is about 80%, and using dosage is rational.
The stability of pharmaceutical preparation is measured
One, test material:
The content of white powdered sample (oxolinic acid is about 1%).
Instrument and medicine: liquid chromatograph instrument system, high speed centrifuge; The microsyringe of 25ul and 100ul; Efficient liquid phase chromatographic analysis Yong oxolinic acid standard substance (purity 〉=99%), dichloromethane, NaOH, dipotassium hydrogen phosphate are analytical pure; Methanol, phosphoric acid are chromatographically pure.
Two, experimental technique:
Chromatographic condition: Alltech C18 reversed phase chromatographic column (15mm * 4mm, granularity 5um); Mobile phase is methanol: phosphoric acid,diluted (volume ratio is 40: 60, pH2.5), filters with 0.45um micropore organic facies filter membrane, and ultrasonic degas.Flow velocity: 1.0mL/min; Detect wavelength: 280nm; Sampling volume: 10uL, retention time is 7～8min; Detection sensitivity 0.01AUFS; Concentration limit 0.010ug/ml.
Standard curve: precision takes by weighing sample 0.0500mg in the 50ml volumetric flask, add a small amount of 0.1mol/LNaOH dissolving after, add mobile phase again and be diluted to graticule.Get 2.50,5.00,7.50,10.00,15.00 respectively, 20.00ml is in the 50ml volumetric flask, be diluted to scale with mobile phase, then the concentration of sample be respectively 0.05,0.10,0.1,0.2,0.3,0.4mg/ml, get the 10ul sample introduction respectively, with peak area (Y) concentration (C) is returned, both linear relationships are good, regression equation: C=24536.58X+9719.24, r=0.999
Sample recovery rate is measured: precision takes by weighing sample 6.670g (by 7.5% production standard, Ying Han oxolinic acid 50mg), puts in the 250ml beaker, adds methanol 20ml.Vibration 10min puts in the tool plug centrifuge tube, and 4 ℃ of high speed centrifuges, the centrifugal 15min of 5000rpm get supernatant, and residue continue to add after methanol 20ml stirs, and continues with handling as stated above, and centrifugal back merges supernatant in 50ml volumetric flask standardize solution.Get the 10ml extracting solution then and put in the 100ml volumetric flask, add mobile phase, behind the 0.45um membrane filtration, get the 10ul sample introduction, the response rate of record peak area and Ji Suan oxolinic acid to graticule.See the following form
| Batch | Theoretical amount (μ g/ml) | Actual measured amount (μ g/ml) | The response rate (%) | Average recovery rate (%) |
| ????1 | ????10.00 | ????9.832 | ???98.32 | ????99.75 |
| ????10.00 | ????9.911 | ???99.11 | ||
| ????2 | ????10.00 | ????9.937 | ???99.37 | |
| ????10.00 | ????9.928 | ???99.28 | ||
| ????3 | ????10.00 | ????10.143 | ???101.43 | |
| ????10.00 | ????10.097 | ???100.97 |
Three, pharmaceutical preparation study on the stability:
The drugs used in aquiculture preparation stability is investigated with reference to the relevant accelerated test of Ministry of Public Health bureau of drug administration " new drug preclinical study guideline compilation " and is carried out.
Since the host of this product Wei oxolinic acid, and adjuvant be high surely vitamin C and starch, because of not producing degradation reactions such as oxidation, hydrolysis between host and adjuvant.Therefore three batches sample is sub-packed in the Brown Glass Brown glass bottles and jars only, place 40 ± 2 ℃, relative humidity is in 75 ± 5% the incubator, every sampling in 5,15,30,60,90 days, the method of pressing in the sample recovery rate mensuration is extracted sample Zhong De oxolinic acid, content with high effective liquid chromatography for measuring Zhi Ji Zhong oxolinic acid the results are shown in following table:
| Batch | Theoretical amount (μ g/ml) | Actual measured amount (μ g/ml) | The 90d meansigma methods | |||||
| ???5d | ??15d | ??30d | ??60d | ??90d | Content (μ g/ml) | The response rate (%) | ||
| ???1 | ??10.00 | ???9.903 | ??9.812 | ??9.743 | ??9.702 | ??9.689 | ??9.800 | ???98.00 |
| ???2 | ??10.00 | ???9.926 | ??9.915 | ??9.874 | ??9.796 | ??9.759 | ||
| ???3 | ??10.00 | ???10.004 | ??9.985 | ??9.967 | ??9.965 | ??9.945 | ||
The average content of active ingredient — oxolinic acid before carrying out accelerated test is 9.975 μ g/ml in three batches the sample, and the average content after 90 days accelerated tests is 9.800 μ g/ml, decreased average 1.77%.Accelerated test result shows that this pharmaceutical preparation stability is better, and it is 2 years that effect duration orders temporarily.
Four, main conclusions:
The average recovery rate of three batches sample is 99.75%, shows content of effective conformance with standard in the sample; At 40 ± 2 ℃, relative humidity is that the average recovery rate of preserving after three months under 75 ± 5% conditions is 98.00%, reduces by 1.77%.Therefore the present invention is comparatively stable, and ordering effect duration temporarily according to its stability is 2 years.
The specific embodiment
A kind of antibacterial for aquatic animal, its Han You oxolinic acid, vitamin C, starch, described vitamin C adopts stable vitamin C usually.The weight proportion of Suo Shu De oxolinic acid is 5%～10%, and described ascorbic weight proportion is 1%～4%, and the weight proportion of described starch is 88%～92%.
Further prescription is: the weight proportion of Suo Shu De oxolinic acid is 7%～8%, and described ascorbic weight proportion is 2%～3%, and the weight proportion of described starch is 89.5%～90.5%.
A kind of preparation method of antibacterial for aquatic animal, it comprises the steps,
(1), with starch in the oven dry 2 hours down of 105 ℃～110 ℃ temperature conditions, 40 mesh sieves are crossed in the cooling back;
(2), Cheng Qu oxolinic acid and vitamin C in proportion;
(3), Qu Yu the starch that oxolinic acid and ascorbic gross weight equate, make starch Yu oxolinic acid, vitamin C fully are mixed and made into female powder;
(4), female powder is fully mixed with remaining starch again.
The purity content of the oxolinic acid that is taken by weighing in step (2) is more than or equal to 98%.
A kind of method of feeding medicine of antibacterial for aquatic animal, it comprises the steps,
(1), in bait, 2% adds this medicine by weight ratio;
(2), by weight ratio about 5% with the bait feeding forage fiss;
(3), 7%～8% forage fiss thrown something and fed aquatic animals such as Siniperca chuatsi by weight ratio.
Siniperca chuatsi need be taken pharmaceutical chemistry when suffering from enteritis, serious gill rot and hemorrhagic disease.But Siniperca chuatsi is the predation live fish, and pharmaceutical chemistry needs earlier to be eaten in the tripe by the feedstuff fish, by the Siniperca chuatsi predation, belongs to indirect drug regimen again.Be generally the 6-7% of fish body weight by feedstuff fish food ration, the amount of Siniperca chuatsi predation feedstuff fish is generally 10% of body weight, designs the prescription prescription by the effective dose of medicine again.So for aquatic animals such as Siniperca chuatsis, the present invention adopts above-mentioned method of feeding medicine.
The major advantage of antibacterial for aquatic animal of the present invention can be by the concrete proof of test, its minimal inhibitory concentration is less, safe in utilization, the Siniperca chuatsi bacterial haemorrhage disease that is caused by Aeromonas there is the better prevention effect, the effective percentage of control is about 80%, and the stability of the effective ingredient of this antibacterial medicine preparation better.
Claims (7)
1, a kind of antibacterial for aquatic animal is characterized in that: its Han You oxolinic acid, vitamin C, starch.
2, antibacterial for aquatic animal according to claim 1 is characterized in that: the weight proportion of Suo Shu De oxolinic acid is 5%～10%, and described ascorbic weight proportion is 1%～4%, and the weight proportion of described starch is 88%～92%.
3, antibacterial for aquatic animal according to claim 2 is characterized in that: the weight proportion of Suo Shu De oxolinic acid is 7%～8%, and described ascorbic weight proportion is 2%～3%, and the weight proportion of described starch is 89.5%～90.5%.
4, antibacterial for aquatic animal according to claim 1 is characterized in that: described vitamin C is stable vitamin C.
5, a kind of preparation method of antibacterial for aquatic animal is characterized in that: it comprises the steps,
(1), with starch in the oven dry 2 hours down of 105 ℃～110 ℃ temperature conditions, 40 mesh sieves are crossed in the cooling back;
(2), Cheng Qu oxolinic acid and vitamin C in proportion;
(3), Qu Yu the starch that oxolinic acid and ascorbic gross weight equate, make starch Yu oxolinic acid, vitamin C fully are mixed and made into female powder;
(4), female powder is fully mixed with remaining starch again.
6, the preparation method of antibacterial for aquatic animal according to claim 5 is characterized in that: the purity content of the oxolinic acid that is taken by weighing in step (2) is more than or equal to 98%.
7, a kind of method of feeding medicine of antibacterial for aquatic animal is characterized in that: it comprises the steps,
(1), in bait, adds this medicine;
(2), with bait feeding living body bait fish;
(3), the living body bait fish is thrown something and fed aquatic animal.
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| CNB2004101032256A CN1299684C (en) | 2004-12-30 | 2004-12-30 | Aquatic animal antibacterial agent and preparation method thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103991643A (en) * | 2014-05-14 | 2014-08-20 | 昆山市富众网络科技有限公司 | Insect expelling type crab box |
| CN105495035A (en) * | 2015-12-04 | 2016-04-20 | 全椒县城西畜牧养殖专业合作社 | Selenium-rich baitfish additive for siniperca chuatsi |
| CN112255342A (en) * | 2020-10-14 | 2021-01-22 | 合肥海关技术中心 | Method for determining residual quantity of prothioconazole and metabolite thioneazole thereof in potatoes |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| NO882653D0 (en) * | 1988-06-15 | 1988-06-15 | Apothekernes Lab | DOSAGE FORM. |
| ATE308983T1 (en) * | 1998-08-25 | 2005-11-15 | Nippon Suisan Kaisha Ltd | USE OF NATURAL PHYSIOLOGICALLY ACTIVE SUBSTANCES AGAINST PARASITIC FISH DISEASES |
-
2004
- 2004-12-30 CN CNB2004101032256A patent/CN1299684C/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103991643A (en) * | 2014-05-14 | 2014-08-20 | 昆山市富众网络科技有限公司 | Insect expelling type crab box |
| CN103991643B (en) * | 2014-05-14 | 2016-03-23 | 昆山市富众网络科技有限公司 | Expelling parasite type crab box |
| CN105495035A (en) * | 2015-12-04 | 2016-04-20 | 全椒县城西畜牧养殖专业合作社 | Selenium-rich baitfish additive for siniperca chuatsi |
| CN112255342A (en) * | 2020-10-14 | 2021-01-22 | 合肥海关技术中心 | Method for determining residual quantity of prothioconazole and metabolite thioneazole thereof in potatoes |
| CN112255342B (en) * | 2020-10-14 | 2023-10-13 | 合肥海关技术中心 | Method for measuring residual quantity of prothioconazole and metabolite thioketoconazole in potatoes |
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| CN1299684C (en) | 2007-02-14 |
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