CN102759622A - Nitrofurans drug metabolite assay kit and production method thereof - Google Patents
Nitrofurans drug metabolite assay kit and production method thereof Download PDFInfo
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Abstract
The invention provides a nitrofurans drug metabolite assay kit, which consists of a furazolidone metabolite test paper strip, a furaltadone metabolite test paper strip, a nitrofurazone metabolite test paper strip and a furantoin metabolite test paper strip. Each test paper strip consists of a sample pad, a colloidal gold pad, a nitrocellulose membrane, a sample adsorbing pad and a PVC (polyvinyl chloride) supporting plate, wherein the colloidal gold layer is a nitrofurans drug metabolite monoclonal antibody polyester film marked with colloidal gold, the nitrocellulose membrane is sequentially coated with nitrofurans drug metabolite coupled carrier protein as a test line (a T-line) and goat anti-mouse IgG antibody as a quality control line (a C-line). The nitrofurans drug metabolite assay kit is produced according to the nano colloidal gold technology, antigen-antibody specificity reactions and principles of immunity competitive inhibition reaction, and is used for testing whether samples contain nitrofurans drug metabolites or not.
Description
Technical field
The present invention relates to the determination techniques field of biology immunization method, particularly relate to a kind of a kind of nitrofurans medicament metabolite detection kit of utilizing colloid gold immune layer fabrication techniques and preparation method thereof.
Background technology
The itrofurans medicine is a kind of spectrum microbiotic; Mainly be meant furaltadone, furazolidone, furantoin, nitrofurazone; Has the excellent antibiotic effect; Be widely used in feed addictive and medicine, be used to prevent and treat the enterogastric diseases of the pig, ox, poultry and the honeybee that cause by salmonella and escherich's bacillus.They have pernicious influence to human health, and carcinogenicity is arranged.European Union classifies the itrofurans microbiotic as category-A forbidding medicine from the consideration of food security in the 96/23/EC rules, decision forbids in animal feed, adding any itrofurans microbiotic from January 1st, 1997.China has also issued in 2002 and has banned use of the antibiotic ban of itrofurans.EU Committee has passed through 2003/181/EC council resolution in 2003, it is 1 μ g/kg that the minimum of having set up the whole bag of tricks of the metabolin that is used for detecting poultry product and aquatic products itrofurans medicine requires the performance limit value.At present, very strict restriction has all been done to nitrofuran antibiotics residue in the import animal food by European Union, the U.S. and other countries, requires 4 kinds of nitrofuran metabolite residues must not surpass 1 μ g/kg.
For the safety of humans and animals is necessary water source and food (as: meat) are carried out the antibiotic residue detection.The detection of itrofurans medicine itself is difficult to, because after eating these medicines, metabolism can take place soon.These metabolic products can exist for a long time in tissue.AOZ is that Furaxone metabolite, AMOZ are that metabolin, the SEM of furaltadone is that Furacilin metabolite, AHD are Cistofuran metabolites.Therefore, can whether contain nitrofurans medicament metabolite in the food to judge the security of food through detecting to human body.
At present, the method for detection nitrofurans medicament metabolite mainly contains high performance liquid chromatography (HPLC), LC/MS (LC-MS), liquid chromatography-tandem mass spectrometry method (LC-MS/MS), enzyme-linked immunosorbent assay (ELISA).These methods are highly sensitive, and the result is accurate, need expensive instrument but have, detects loaded down with trivial details, time-consuming, the shortcomings such as professional training that testing staff's needs are certain.
The present invention adopts the colloidal gold immunochromatographimethod technology to prepare a kind of nitrofurans medicament metabolite detection kit.That the present invention has is simple to operate, it is quick, highly sensitive to detect, need not characteristics such as complex instrument.
Summary of the invention
The object of the present invention is to provide a kind of simple, quick, highly sensitive, nitrofurans medicament metabolite detection kit that cost is low and preparation method thereof.
The nitrofurans medicament metabolite detection kit is assembled by metabolin test strip, Furacilin metabolite test strip, the Cistofuran metabolite test strip of Furaxone metabolite test strip, furaltadone; Every kind of reagent strip is made up of sample pad, collaurum pad, nitrocellulose filter, suction appearance pad and PVC back up pad, and tight adhesion has sample pad, collaurum pad, nitrocellulose filter and suction appearance pad successively on the PVC back up pad.Said sample pad is a spun glass; The nitrofurans medicament metabolite monoclonal antibody polyester film that said collaurum pad is a colloid gold label; Be coated with detection line (T line) and nature controlling line (C line) on the said nitrocellulose filter successively, wherein encapsulated nitrofurans medicament metabolite coupling carrier albumen on the detection line (T line), encapsulated sheep anti-mouse igg antibody on the nature controlling line (C line); Said suction appearance pad is thieving paper.
The carrier protein of coupling nitrofurans medicament metabolite is bovine serum albumin(BSA) (BSA) or oralbumin (OVA).
The present invention adopts nano-colloid technology for gold and antigen and antibody specific reaction; The principle of using immunity competition inhibitory reaction is prepared from; The reaction of the nitrofurans medicament metabolite antibody of the nitrofurans medicament metabolite coupling carrier protein competition association colloid gold mark that encapsulates through detection line (T line) on the nitrofurans medicament metabolite that possibly contain in the sample to be checked and the nitrocellulose filter makes the T line manifest naked eyes red color visible band and is used for judging whether sample to be checked contains nitrofurans medicament metabolite.
The invention provides a kind of preparation method of nitrofurans medicament metabolite detection kit, may further comprise the steps:
(1) preparation nitrofurans medicament metabolite coupling carrier albumen
The itrofurans medicine is a small-molecule substance, has only immunoreactivity, does not have immunogenicity, can not bring out body and produce immune response, must with the macromolecular carrier albumen coupling after just can have immunogenicity.Therefore adopt active ester method that nitrofurans medicament metabolite haptens and carrier protein couplet are obtained nitrofurans medicament metabolite coupling carrier albumen as immunogene and coating antigen.Wherein carrier protein is bovine serum albumin(BSA) (BSA) or oralbumin (OVA).
(2) preparation nitrofurans medicament metabolite monoclonal antibody
Adopt nitrofurans medicament metabolite coupling carrier albumen as the immunogen immune BALB/C mice, preparation nitrofurans medicament metabolite monoclonal antibody.
(3) preparation collaurum
Get 0.01% aqueous solution of chloraurate, heated and boiled.Add 1% citric acid, three sodium water solutions as required rapidly, continue to boil about 5min, occur orange red.The size of the colloid gold particle of processing like this is 20-40nm.
(4) preparation collaurum pad
Get the colloidal gold solution that grain size is 20-40nm, use 0.2mol/L K
2CO
3The pH value of colloidal gold solution is transferred to 8.0-9.0, and room temperature was placed 30 minutes; In above-mentioned solution, add the nitrofurans medicament metabolite monoclonal antibody, the concentration that makes the nitrofurans medicament metabolite monoclonal antibody is 10-40 μ g/ml collaurum, and after mixing, room temperature was placed 30 minutes; Add 10%BSA solution then, the concentration that makes BSA solution is 10-30 μ l/ml collaurum, mixes, and room temperature was placed 10 minutes; Above-mentioned solution left the heart 30 minutes in 12000, carefully drew supernatant, discarded, and remaining deposition is redissolved the liquid dissolving with the collaurum of 3070% initial collaurum volume, obtains nitrofurans medicament metabolite monoclonal antibody colloid gold label thing; Nitrofurans medicament metabolite monoclonal antibody-colloid gold label thing is pressed 15-30 μ l/cm
2Ratio be layered on uniformly on the polyester film, put into 45 ℃ of drying boxes dry 2.5-3 hour, process the collaurum pad.
Above-mentioned collaurum redissolves liquid for containing 0.30%Tris, 5% sucrose, 0.50%PVP, 0.30%Casein-Na, the solution of pH value 8.0-9.0.
(5) encapsulate nitrofurans medicament metabolite coupling carrier albumen, sheep anti-mouse igg antibody
The NC film is sticked on the PVC plate, and thieving paper is attached to and pushes down film 1-1.5mm on the PVC plate.Open and draw the film appearance, press cancel key and clean a stroke film appearance, being provided with and drawing a film instrument parameter is C line 1.0 μ l/cm, T line 1.0 μ l/cm;
The PVC plate that posts is placed on stroke film appearance, on the NC film, draws C, T line respectively with C, T line coating buffer.Draw the red marking pen of the underproof usefulness of film and mark, will encapsulate plate and put drying box and be provided with 45 ℃, dry 1-1.5 hour.
Wherein, T line coating buffer is the nitrofurans medicament metabolite coupling carrier albumen of 0.8-4.0mg/ml; C line coating buffer is the sheep anti-mouse igg antibody of 0.6-2.0mg/ml.
(6) processing of sample pad
Spun glass is soaked 20-40min in the PBS of 0.01M pH 7.0-8.0, wherein contain 0.5-1.5%BSA in the PBS, 0.5-1.0%Tweeen-20,38 ℃ of oven dry in drying baker are preserved, and are subsequent use
(7) assembling kit
Get the above-mentioned collaurum pad for preparing, PVC plate and sample pad; The collaurum pad is cut into the stripe shape that width is 10mm, then the collaurum pad be affixed on NC film on the PVC plate near the T line end, press NC film 1-1.5mm; Sample pad is affixed on the top of collaurum pad, exposes gold pad 2-3mm (as shown in Figure 1).Open cutting cutter, cut width, the above-mentioned PVC plate that assembles is put on the cutting cutter, carry out slitting, obtain the said test strips that is used to detect nitrofurans medicament metabolite by width is set by the product requirement setting; Test strips is packed in the plastic clip, is assembled into kit.
The detection method of kit according to the invention is: with the test sample balance to room temperature; Take out the nitrofurans medicament metabolite detection kit, horizontal positioned; In sample pad, add 2-3 and drip sample, observe and write down the colour developing situation of C, T line in the time of 5-10 minute, judge testing result; Or with after about 10 seconds in the terminal immersion of the nitrofurans medicament metabolite test strip sample pad sample solution, horizontal positioned; Observe and write down the colour developing situation of C, T line in the time of 5-10 minute, judge testing result.
Kit of the present invention adopts colloidal gold immunochromatographimethod technical measurement nitrofurans medicament metabolite; During test sample; Because of the capillary action sample solution along test strips to inhaling appearance pad one an end chromatography; If contain nitrofurans medicament metabolite in the sample; They will and detection line (T line) on limited antibody combining site on the nitrofurans medicament metabolite monoclonal antibody of the nitrofurans medicament metabolite coupling carrier protein competition association colloid gold mark that encapsulates; When the nitrofurans medicament metabolite in the sample reached finite concentration, also saturated fully with the nitrofurans medicament metabolite monoclonal antibody generation immune response of colloid gold label, this moment, colloidal gold composite did not have the nitrofurans medicament metabolite coupling carrier protein combination that encapsulates on vacant site and the detection line; This moment, the T line did not develop the color this positive result; If do not contain nitrofurans medicament metabolite in the sample; Mark the colloid gold particle of nitrofurans medicament metabolite monoclonal antibody will be to the T line position in company with the sample chromatography; Immune association reaction takes place with the nitrofurans medicament metabolite coupling carrier albumen that encapsulates on the T line; Colloid gold particle is piled up at the T line position and is made the T line demonstrate a macroscopic red stripes, this negative result.No matter whether contain nitrofurans medicament metabolite in the sample; The colloid gold label thing all can form a macroscopic red stripes with the sheep anti-mouse igg reaction that the C line encapsulates; Whether the C line develops the color; Be to judge whether enough samples are arranged, whether normal chromatography process standard simultaneously also as the inner quality standard of reagent.
Description of drawings
Fig. 1 nitrofurans medicament metabolite detection kit synoptic diagram;
The structural representation of test strips in Fig. 2 nitrofurans medicament metabolite detection kit;
The reference numeral explanation:
1: sample pad;
2: collaurum pad (polyester film of colloid gold label nitrofurans medicament metabolite monoclonal antibody)
3: nitrocellulose filter (T: the detection line that has encapsulated nitrofurans medicament metabolite coupling carrier albumen; C: the nature controlling line that has encapsulated sheep anti-mouse igg antibody);
4: inhale the appearance pad;
The 5:PVC back up pad;
The testing result synoptic diagram of Fig. 3 kit of the present invention.
Be followed successively by line positive test symbol of C line from left to right; T, two line negative result of C; Invalid.
Embodiment 1: the preparation of nitrofurans medicament metabolite detection kit
1. prepare nitrofurans medicament metabolite coupling BSA
Adopt active ester method respectively AOZ, AMOZ, SEM and AHD haptens and bovine serum albumin(BSA) coupling to be obtained AOZ-BSA conjugate, AMOZ-BSA conjugate, SEM-BSA conjugate and AHD-BSA conjugate as immunogene and coating antigen.
2. prepare the nitrofurans medicament metabolite monoclonal antibody
The AOZ-BSA conjugate prepares the AOZ monoclonal antibody as the immunogen immune BALB/C mice; The AMOZ-BSA conjugate prepares the AMOZ monoclonal antibody as the immunogen immune BALB/C mice; The SEM-BSA conjugate prepares the SEM monoclonal antibody as the immunogen immune BALB/C mice; The AHD-BSA conjugate prepares the AHD-monoclonal antibody as the immunogen immune BALB/C mice.。
3. preparation collaurum
Get 0.01% aqueous solution of chloraurate 100mL, heated and boiled.Add 1% citric acid, three sodium water solution 0.75mL as required rapidly, continue to boil about 5min, occur orange red.The size of the colloid gold particle of processing like this is 20-40nm.
4. prepare the collaurum pad
Get the centrifuge tube of 4 10ml, number 1,2,3,4 respectively, in 4 centrifuge tubes, add the colloidal gold solution that the 5ml grain size is 20-40nm respectively, with 0.2mol/L K2CO3 the pH value of colloidal gold solution is transferred to 9.0, room temperature was placed 30 minutes; In No. 1 centrifuge tube, adding concentration is the AOZ monoclonal antibody 39.5 μ l of 3.8mg/ml; In No. 2 centrifuge tubes, adding concentration is the AMOZ monoclonal antibody 40 μ l of 3.5mg/ml; In No. 3 centrifuge tubes, adding concentration is the SEM monoclonal antibody 37.5 μ l of 4.0mg/ml, and in No. 4 centrifuge tubes, adding concentration is the AHD monoclonal antibody 39.5 μ l of 3.8mg/ml; After mixing respectively, room temperature was placed 30 minutes; In the 1-4 centrifuge tube, all add the 10%BSA solution of 100 μ l then, mix, room temperature was placed 10 minutes; Above-mentioned solution left the heart 30 minutes in 12000; The careful supernatant of drawing; Discard; Remaining deposition all with the collaurum redissolution liquid dissolving of 2.5ml, obtains AOZ monoclonal antibody-colloid gold label thing, AMOZ monoclonal antibody-colloid gold label thing, SEM monoclonal antibody-colloid gold label thing and AHD monoclonal antibody-colloid gold label thing.Above-mentioned four kinds of colloid gold label things are pressed 20 μ l/cm respectively
2Ratio be layered on uniformly on 4 polyester films, put into 45 ℃ of drying boxes dry 3 hours, process the collaurum pad.
Above-mentioned collaurum redissolves liquid for containing 0.30%Tris, 5% sucrose, 0.50%PVP, 0.30%Casein-Na, the solution of pH value 9.0.
5. encapsulate nitrofurans medicament metabolite coupling BSA, sheep anti-mouse igg antibody
The NC film is sticked on the PVC plate, and thieving paper is attached to and pushes down film 1-1.5mm on the PVC plate.Open and draw the film appearance, press cancel key and clean a stroke film appearance, being provided with and drawing a film instrument parameter is C line 1.0 μ l/cm, T line 1.0 μ l/cm;
The PVC plate that posts is placed on stroke film appearance, is AOZ-BSA conjugate, AMOZ-BSA conjugate, SEM-BSA conjugate and AHD-BSA conjugate with T line coating buffer respectively, and C line coating buffer is that sheep anti-mouse igg antibody is drawn C, T line respectively on the NC film.Draw the red marking pen of the underproof usefulness of film and mark, will encapsulate plate and put drying box and be provided with 45 ℃, dry 1.5 hours.
Wherein, T line coating buffer AOZ-BSA conjugate concentration is that 2.0mg/ml, AMOZ-BSA conjugate concentration are that 1.0mg/ml, SEM-BSA conjugate concentration are that 1.6mg/ml, AHD-BSA conjugate concentration are 2.5mg/ml; C line coating buffer sheep anti-mouse igg antibody concentration is 1.0mg/ml.
6. the processing of sample pad
Spun glass is soaked 20-40min in the PBS of 0.01M pH 8.0, wherein contain 1.0%BSA in the PBS, 0.5%Tweeen-20,38 ℃ of oven dry in drying baker are preserved, and are subsequent use
7. assembling kit
Get the above-mentioned collaurum pad for preparing, PVC plate and sample pad; The collaurum pad is cut into the stripe shape that width is 10mm; Then with the collaurum pad of AOZ monoclonal antibody-colloid gold label thing preparation be affixed on NC film on the PVC plate that T line coating buffer is the AOZ-BSA conjugate near the T line end; Press NC film 1-1.5mm, sample pad is affixed on the top of collaurum pad, exposes gold pad 2-3mm; Open cutting cutter, being provided with and cutting width is 3.8mm, and the above-mentioned PVC plate that assembles is put on the cutting cutter, carries out slitting by width is set, and obtains the Furaxone metabolite test strip.
Press NC film 1-1.5mm with what the collaurum pad of AMOZ monoclonal antibody-colloid gold label thing preparation was affixed on NC film on the PVC plate that T line coating buffer is the AMOZ-BSA conjugate near the T line end, sample pad is affixed on the top of collaurum pad, exposes gold pad 2-3mm; Open cutting cutter, being provided with and cutting width is 3.8mm, and the above-mentioned PVC plate that assembles is put on the cutting cutter, carries out slitting by width is set, and obtains the metabolin test strip of furaltadone.
With the collaurum pad of SEM monoclonal antibody-colloid gold label thing preparation be affixed on NC film on the PVC plate that T line coating buffer is the SEM-BSA conjugate near the T line end, press NC film 1-1.5mm, sample pad is affixed on the top of collaurum pad, exposes gold pad 2-3mm; Open cutting cutter, being provided with and cutting width is 3.8mm, and the above-mentioned PVC plate that assembles is put on the cutting cutter, carries out slitting by width is set, and obtains the Furacilin metabolite test strip.
With the collaurum pad of AHD monoclonal antibody-colloid gold label thing preparation be affixed on NC film on the PVC plate that T line coating buffer is the AHD-BSA conjugate near the T line end, press NC film 1-1.5mm, sample pad is affixed on the top of collaurum pad, exposes gold pad 2-3mm; Open cutting cutter, being provided with and cutting width is 3.8mm, and the above-mentioned PVC plate that assembles is put on the cutting cutter, carries out slitting by width is set, and obtains the Cistofuran metabolite test strip.
Above-mentioned 4 kinds of test strips that prepare are assembled in the plastic clip in order, are prepared into the nitrofurans medicament metabolite detection kit.
Embodiment 2: the detection of nitrofurans medicament metabolite detection kit
1. the processing of sample
After getting 1g tissue sample tissue sample and smashing to pieces, add 5ml distilled water, benzaldehyde (ethanolic solution) the 100 μ l of the HCl 0.5ml of 1M and 0.01M fully vibrate, and place 38 ℃ to spend the night.The K that adds 0.1M then
2HPO
45ml, the NaOH 0.4ml of 1M and the ethyl acetate of 5ml, thermal agitation 30 seconds.4000rpm is centrifugal 10 minutes under the room temperature, gets 2.5ml ethyl acetate supernatant then and transfers in the new container.Under 45 ℃, with nitrogen it is dried up, (pH 7.4,0.05%Tween-20) mixing with the 0.01M PBST of 1ml to dissolve the back again with normal hexane then.4000rpm is centrifugal 10 minutes under the room temperature, gets bottom aqueous phase at last and measures.
2. detection method:
Take out the nitrofurans medicament metabolite detection kit, horizontal positioned; On sample pad, splash into 3 samples, observe and write down the colour developing situation of C, T line after 10 minutes, judge testing result.
3. the result judges
Positive: the T line does not develop the color, and only C line colour developing is judged to be positive findings;
Negative: T line, C line all develop the color, and are judged to be negative findings;
Invalid: the C line does not develop the color, and the rotten damage of maloperation or kit is described.
Claims (7)
1. nitrofurans medicament metabolite detection kit and preparation method thereof is characterized in that being assembled by metabolin test strip, Furacilin metabolite test strip, the Cistofuran metabolite test strip of Furaxone metabolite test strip, furaltadone.
2. described nitrofurans medicament metabolite detection kit of claim 1 and preparation method thereof; It is characterized in that the test strips that is comprised is made up of sample pad, collaurum pad, nitrocellulose filter, suction appearance pad and PVC back up pad, tight adhesion has sample pad, collaurum pad, nitrocellulose filter and suction appearance pad successively on the PVC back up pad.Said sample pad is a spun glass; The nitrofurans medicament metabolite monoclonal antibody polyester film that said collaurum pad is a colloid gold label; Be coated with detection line (T line) and nature controlling line (C line) on the said nitrocellulose filter successively, wherein encapsulated nitrofurans medicament metabolite coupling carrier albumen on the detection line (T line), encapsulated sheep anti-mouse igg antibody on the nature controlling line (C line); Said suction appearance pad is thieving paper.
3. described nitrofurans medicament metabolite detection kit of claim 1 and preparation method thereof is characterized in that described nitrofurans medicament metabolite coupling carrier albumen is nitrofurans medicament metabolite coupling bovine serum albumin(BSA) or oralbumin; Described nitrofurans medicament metabolite monoclonal antibody is obtained as the immunogen immune BALB/C mice by nitrofurans medicament metabolite coupling carrier albumen.
4. described nitrofurans medicament metabolite detection kit of claim 1 and preparation method thereof is characterized in that described collaurum is by gold chloride (HAuCl
4) under the effect of reductive agent citric acid trisodium, process size and be the colloid gold particle of 20-40nm.
5. described nitrofurans medicament metabolite detection kit of claim 1 and preparation method thereof is characterized in that the preparation method of described collaurum pad is: get the colloidal gold solution that grain size is 20-40nm, use 0.2mol/L K
2CO
3The pH value of colloidal gold solution is transferred to 8.0-9.0, and room temperature was placed 30 minutes; In above-mentioned solution, add the nitrofurans medicament metabolite monoclonal antibody, the concentration that makes the nitrofurans medicament metabolite monoclonal antibody is 10-40 μ g/ml collaurum, and after mixing, room temperature was placed 30 minutes; Add 10%BSA solution then, the concentration that makes BSA solution is 10-30 μ l/ml collaurum, mixes, and room temperature was placed 10 minutes; Above-mentioned solution left the heart 30 minutes in 12000, carefully drew supernatant, discarded, and remaining deposition is redissolved the liquid dissolving with the collaurum of the initial collaurum volume of 30-70%, obtains nitrofurans medicament metabolite monoclonal antibody-colloid gold label thing; Nitrofurans medicament metabolite monoclonal antibody-colloid gold label thing is pressed 15-30 μ l/cm
2Ratio be layered on uniformly on the polyester film, put into 45 ℃ of drying boxes dry 2.5-3 hour, process the collaurum pad.
6. described nitrofurans medicament metabolite detection kit of claim 1 and preparation method thereof; It is characterized in that described collaurum redissolves liquid for containing 0.30%Tris, 5% sucrose, 0.50%PVP; 0.30%Casein-Na, the solution of pH value 8.0-9.0.
7. described nitrofurans medicament metabolite detection kit of claim 1 and preparation method thereof; The method for coating that it is characterized in that T on the described nitrocellulose filter, C line is: the NC film is sticked on the PVC plate, and thieving paper is attached to and pushes down film 1-1.5mm on the PVC plate.Open and draw the film appearance, press cancel key and clean a stroke film appearance, being provided with and drawing a film instrument parameter is C line 1.0 μ l/cm, T line 1.0 μ l/cm; The PVC plate that posts is placed on stroke film appearance, on the NC film, draws C, T line respectively with C, T line coating buffer.Draw the red marking pen of the underproof usefulness of film and mark, will encapsulate plate and put drying box and be provided with 45 ℃, dry 1-1.5 hour.Wherein, T line coating buffer is the nitrofurans medicament metabolite coupling carrier albumen of 0.8-4.0mg/ml; C line coating buffer is the sheep anti-mouse igg antibody of 0.6-2.0mg/ml.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104374910A (en) * | 2014-11-19 | 2015-02-25 | 无锡中德伯尔生物技术有限公司 | Kit and method applied to detection of nitrofurans drug metabolite |
| CN104880523A (en) * | 2015-04-28 | 2015-09-02 | 衢州出入境检验检疫局综合技术服务中心 | Method for determining nitrofuran metabolites in bee wax through high performance liquid chromatography tandem mass spectrometry |
| CN112578110A (en) * | 2020-12-16 | 2021-03-30 | 西北农林科技大学 | Biological probe and kit for detecting furacilin and furazolidone and application |
| CN113960013A (en) * | 2021-11-08 | 2022-01-21 | 北京同仁堂健康（大连）海洋食品有限公司 | Method for detecting nitrofuran drug metabolites based on Br-assisted SERS |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004008188A2 (en) * | 2002-07-12 | 2004-01-22 | Affymetrix, Inc. | System and method for examination of microarrays using scanning electron microscope |
| CN1793927A (en) * | 2005-12-31 | 2006-06-28 | 江南大学 | Reagent box for detecting clenbuterol hydrochloride and its detection method |
| CN201004063Y (en) * | 2006-09-01 | 2008-01-09 | 河南省动物免疫学重点实验室 | Quick testing test paper sheet and test paper card for furazolidone metabolized material |
| CN101320037A (en) * | 2008-06-25 | 2008-12-10 | 江南大学 | Preparation method of colloidal gold chromatography test paper for fast detecting nitrofuran metabolite |
| CN101320038A (en) * | 2008-06-25 | 2008-12-10 | 江南大学 | Preparation method of colloidal gold chromatography test paper for fast detecting urolong metabolite |
-
2011
- 2011-04-28 CN CN2011101084778A patent/CN102759622A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004008188A2 (en) * | 2002-07-12 | 2004-01-22 | Affymetrix, Inc. | System and method for examination of microarrays using scanning electron microscope |
| CN1793927A (en) * | 2005-12-31 | 2006-06-28 | 江南大学 | Reagent box for detecting clenbuterol hydrochloride and its detection method |
| CN201004063Y (en) * | 2006-09-01 | 2008-01-09 | 河南省动物免疫学重点实验室 | Quick testing test paper sheet and test paper card for furazolidone metabolized material |
| CN101320037A (en) * | 2008-06-25 | 2008-12-10 | 江南大学 | Preparation method of colloidal gold chromatography test paper for fast detecting nitrofuran metabolite |
| CN101320038A (en) * | 2008-06-25 | 2008-12-10 | 江南大学 | Preparation method of colloidal gold chromatography test paper for fast detecting urolong metabolite |
Non-Patent Citations (4)
| Title |
|---|
| J.V.SAMSONOVA ET AL: "The identification of potential alternative biomarkers of nitrofurazone abuse in animal derived food products", 《FOOD AND CHEMICAL TOXICOLOGY》 * |
| J.V.SAMSONOVA ET AL: "The identification of potential alternative biomarkers of nitrofurazone abuse in animal derived food products", 《FOOD AND CHEMICAL TOXICOLOGY》, vol. 46, 31 December 2008 (2008-12-31), pages 1548 - 1554 * |
| 向军俭等: "胶体金免疫层析法检测食用肉中的氨基脲", 《食品科学》 * |
| 朱海等: "呋喃唑酮代谢物荧光纳米颗粒免疫层析法的建立", 《畜牧与饲料科学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104374910A (en) * | 2014-11-19 | 2015-02-25 | 无锡中德伯尔生物技术有限公司 | Kit and method applied to detection of nitrofurans drug metabolite |
| CN104880523A (en) * | 2015-04-28 | 2015-09-02 | 衢州出入境检验检疫局综合技术服务中心 | Method for determining nitrofuran metabolites in bee wax through high performance liquid chromatography tandem mass spectrometry |
| CN112578110A (en) * | 2020-12-16 | 2021-03-30 | 西北农林科技大学 | Biological probe and kit for detecting furacilin and furazolidone and application |
| CN113960013A (en) * | 2021-11-08 | 2022-01-21 | 北京同仁堂健康（大连）海洋食品有限公司 | Method for detecting nitrofuran drug metabolites based on Br-assisted SERS |
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