CN105137068A - Florfenicol on-site test paper, and preparation and use methods thereof - Google Patents
Florfenicol on-site test paper, and preparation and use methods thereof Download PDFInfo
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Abstract
The invention particularly relates to florfenicol on-site test paper, and preparation and use methods thereof and belongs to the field of immunology. The test paper includes anti-florfenicol amine secreted from hybridoma FFA-C which is assigned the accession number CCTCC-C201575, a monoclonal antibody C marked by colloidal gold and a carrier plate having adhesive sticker, wherein a sample adding end water absorption layer and a hand holding end water absorption layer are formed at two ends of the carrier plate respectively. A test layer is arranged on the middle of the carrier plate. A glass fiber layer block, on which the monoclonal antibody C marked by colloidal gold is supported, is arranged at the boundary between the test layer and the sample adding end water absorption layer. One end of the glass fiber layer block is arranged under the sample adding end water absorption layer and the other end of the glass fiber layer block is arranged above the test layer. A test line and a quality control line are formed on the test layer extended from the glass fiber layer block, wherein the test line and the quality control line are respectively coated by an anti-florfenicol amine polyclonal antibody and sheep-anti mouse IgG. The test paper quickly, simply and accurately detects the florfenicol and the metabolite, florfenicol amine, thereof on site, and can be used for crude screening test of drug residue of the florfenicol.
Description
Technical field
The present invention relates to the improvement of microbiotic detection technique, be specifically related to Site Detection test strips and preparation thereof, using method, belong to field of immunology.
Background technology
Florfenicol (Florfenico, FF) Chinese: Fluprofen, Florfenicol, chemical name: d (+)-Su-1-p-methylphenyl-2-dichloro acetamino-3-fluorine propyl alcohol, it is the broad spectrum antibiotic of the special chloromycetin of a kind of beasts succeeded in developing by U.S. Schering-Plough in the later stage 1980s, it is third generation chloromycetin series antibiotics, this medicine nineteen ninety goes on the market in Japan first, as the substitute of new chloromycetin, the combination of main suppression 50s subunit thus the activity of anti-bacteria acyltransferase polypeptide transaminase, it is novel resisting gram-positive bacteria, the broad-spectrum antibiotic of Gram-negative bacteria and Thiamphenicol drug-fast bacteria, can be used for treating pig, ox, the bacteriosis of fowl and aquiculture animal.
Chloromycetin (Chloramphenicol, CAP) as first generation chloromycetin series antibiotics, its structural-NO2 group is relevant with suppression hematopoiesis function, therefore CAP has spinal cord hematopoiesis function suppression toxicity, reversible haemocyte can be caused to reduce, and the probability causing irreversible alpastic anemia is 1:30000.FF-CH3SO2 group instead of-NO2 group, dangerous without potential alpastic anemia.The antibody-resistant bacterium of FF to resistance to chloromycetin and Thiamphenicol (Thiamphenicol, TAP) still has antibacterial action simultaneously.Florfenicol plays biological effect as targeted drug, the little instant effect of dosage, Increased Plasma Half-life, and blood concentration is high, can maintain blood medicine for a long time, be widely used in the prevention and corntrol of Animal diseases.Within 2000, China's approval Florfenicol is national two class novel chiral synthon, and aquaculture can be used for treatment common eel Ai Dehuashi disease and red fin fish disease.
FF metabolism is very fast in animal body, main metabolites is Florfenicol oxamic acid, florfenicol amine (Florenicolamine, FFA), Florfenicol alcohol etc., but FFA is topmost metabolic product in most of edible animal tissue, and therefore FFA is by the mark residue calculated as the off-drug period.Although Guo Guifang etc. think that FF parenteral solution is without obvious acute toxicity and subchronic toxicity, but there are some researches show high dose FF can mouse thymus, spleen weight reduce, external research also shows that FF can suppress humoral immunity and the cellular immunity of mouse, the conclusion that this Guo Gui virtue draws is confirmed mutually, shows that FF has stronger immunotoxicity.The extensive application of FF in aquaculture can lead its remaining in animal food, in order to protect public health, China specifies the maximum residue limit of Florfenicol in animal tissue, and the muscle residue limits of requirement is 200 μ g/Kg, liver residue limits is 3000 μ g/Kg, kidney residue limits is 300 μ g/Kg.
Summary of the invention
The present invention is directed to the present situation of fluoride protector, and it is to the harm of human body, design invention provide a kind of can the Test paper of on-the-spot quick, easy, accurate detection Florfenicol and its metabolic product florfenicol amine and preparation thereof, using method.
Technical scheme of the present invention realizes as follows
Florfenicol Site Detection test paper, its special character is to comprise: the anti-florfenicol amine of preserving number secreted by CCTCC-C201575 hybridoma FFA-C, the monoclonal antibody C (being called for short: gold mark monoclonal antibody C) of colloid gold label, anti-florfenicol amine polyclonal antibody (being called for short: resist) and goat anti-mouse igg (being called for short: sheep anti-mouse igg) more, with the carrier board 3 of adhesive sticker, at the both ends of carrier board 3, be respectively equipped with application of sample end water accepting layer 1 and handheld terminal water accepting layer 7, the detection layers 6 be made up of nitrocellulose membrane is provided with at this carrier board 3 middle part, be provided with at this detection layers 6 and application of sample end water accepting layer 1 intersection and be loaded with the glass layer block 2 that gold marks monoclonal antibody C, one end of glass layer block 2 is arranged under application of sample end water accepting layer 1, the other end is arranged on detection layers 6, the detection layers 6 continued with glass layer block 2 is provided with detection line 4 and nature controlling line 5, detection line 4 and nature controlling line 5 place are coated with many anti-and sheep anti-mouse iggs respectively,
Described gold mark monoclonal antibody C prepares by alkali hydrolysis method florfenicol amine-bovine serum albumin(BSA) (FFA-BSA) comlete antigen that florfenicol amine (FFA) haptens and bovine serum albumin(BSA) (BSA) synthesized by glutaraldehyde method coupling, it can be used as immunogene and the anti-florfenicol amine monoclonal antibody prepared, cause after colloid gold label;
Described anti-florfenicol amine polyclonal antibody: be prepare by alkali hydrolysis method the rabbit anteserum that florfenicol amine (FFA) haptens and oralbumin (OVA) prepared as immunogen immune rabbit by glutaraldehyde method coupling complete antigen synthesis (FFA-OVA).
The preparation process of the monoclonal antibody C of described colloid gold label is as follows:
1, prepare colloidal gold solution with the preparation technology of known collaurum, the grain size of obtained collaurum is 10-20nm;
2, by monoclonal antibody C (0.5-1g/L antibody protein), join in above-mentioned 100ml colloidal gold solution by 150-500 μ l, mix even slowly;
3, in mixed solution, add PEG20000, make its final concentration be 1-5%, hold over night;
4, centrifugal 8000-10000 rev/min, 1-1.5 hour, abandon supernatant;
5, get centrifugation, hang with the 0.01mol/L phosphate buffer containing 1-5%PEG20000,0.05-0.25% Tween-20,0.02% Sodium azide, make gold mark monoclonal antibody C;
The pH value of described phosphate buffer is 7.4, wherein containing KCI0.2g, NaCI8.0g, KH
2pO
40.2g, Na
2hPO
412H
2o2.9g, distilled water 1000ml;
In described step 2, monoclonal antibody C and colloidal gold solution mix even 30-60min slowly.
The using method of Florfenicol Site Detection test paper, its special character is to comprise the following steps:
The application of sample end water accepting layer 1 of this test paper is inserted or drips detected sample thereon, 5-10min is being positioned at the detection line 4 of detection layers 6 lower end and is being positioned at the nature controlling line 5 place display redness of detection layers 6 upper end, i.e. two red line, representing residual in sample has Florfenicol or florfenicol amine; Detection line place is without red line, and represent that sample represents and can't detect Florfenicol, florfenicol amine or both concentration lower than 10ug/ml, the display of nature controlling line place is red, represents that this test paper is effective.
The invention has the advantages that: the present invention is with the actual features of prior art, have developed a kind of fluoride protector rapid field Test paper and preparation thereof, using method, adopt animal blood or the physiological saline as appropriate in liver adds of internal organ to pulverize and be prepared into detection sample liquid, it is detected, principle adopts sandwich immunoassay chromatographic theory, namely by the same antigen-reactive detected in sample liquid of gold mark monoclonal antibody C, formed by gold mark monoclonal antibody C-Florfenicol or Florfenicol amine compound, when this compound chromatography to nitrocellulose filter being coated in advance the anti-florfenicol amine polyclonal antibody on detection line (FFA is how anti-) place, antibody herein can identify antigen in this compound, the result of reaction just defines gold mark monoclonal antibody C+ Florfenicol or the how anti-sandwich structure of florfenicol amine+FFA, finally that the colloid gold particle that monoclonal antibody C marks is fixed at this and accumulates the macroscopic red line of appearance, unreacted gold mark monoclonal antibody C then still continues chromatography and moves ahead, when the point of arrival is coated on nature controlling line sheep anti-mouse igg place in advance, Antibody types is that the golden standard gold mark monoclonal antibody C of IgG is combined with by it, thus also occur herein being fixed by colloid gold particle and accumulating and show macroscopic red line, result is: detection line display redness indicates Florfenicol or florfenicol amine remains, i.e. medicine setting sun, detection line does not show redness, represents and can't detect Florfenicol, florfenicol amine or both concentration lower than 10ug/ml, i.e. the residual feminine gender of medicine.Nature controlling line display is red, represents that test paper is effective; Namely or golden labeling antibody C or sheep anti-mouse igg inactivation nature controlling line place does not show redness, illustrates that test paper lost efficacy, and, testing result is invalid.Feature of the present invention is: 1, Florfenicol Site Detection test paper of the present invention, from the cultivation needs of reality, by the cultivation site that the detection of medicament residue is moved to, without the need to specialized facilities and operative technique during use, be applicable to common raiser's cultivation site to detect, application is simple.2, have and detect the feature such as quick, easy, accurate, sensitive, and there is higher stability, just testing result can be shown within 5-10min, once detect, exceed standard can by how foster for cultivated animals a period of time, allow drug metabolism until residual lower than national standard, can go on the market safely, be a kind of method solving the residual problem of medicine of most convenient, decreases and caused huge loss because medicament residue to exceed standard because medicine is residual to the injury of human body and raiser by the return of goods.Improve because medicine remains censorship trouble, the cycle is long, and high reason of charging causes more than 99% raiser to be the situation that not censorship medicine remains before cultivated animals listing; 3, this test paper prepare anti-florfenicol amine how anti-time, it is resisting of preparing with the comlete antigen immunizing rabbit of florfenicol amine and ovalbumin coupling more, this fundamentally avoids the cross reaction with bovine serum albumin(BSA), further ensures the specificity (because in monoclonal antibody preparation, immunogene has bovine serum albumin(BSA) to be carrier protein) of test paper; 4, in whole colloid gold label and test paper preparation process, the bovine serum albumin(BSA) preventing non-specific binding and play sealing process is played, all replaced with the PEG20000 of debita spissitudo, stop the cross reaction caused because of bovine serum albumin(BSA) so further, ensure that the specificity of kit.
Accompanying drawing explanation
Fig. 1: the structural representation of Florfenicol Site Detection test paper of the present invention;
Fig. 2: the testing result schematic diagram of Florfenicol Site Detection test paper of the present invention.
Hybridoma cell strain FFA-C, its deposit number is: CCTCC-C201575; Depositary institution's full name and abbreviation: China typical culture collection center (CCTCC); Depositary institution address: Luojiashan, Wuchang, Wuhan City, Hubei Province Wuhan University preservation center; Preservation date: on May 21st, 2015.
Embodiment
Embodiment 1.
The synthesis of florfenicol amine: be 1:1 according to mol ratio, take FF10.74, g NaOH 2.56g adds 15ml distilled water and dissolves, and heating is stirred to dissolves completely, and now solution is claret clarification, add 6.0g sodium chloride again, be stirred to and dissolve completely, with thin-layer chromatography plate layer chromatography mixed liquor after cooling, with the stifling colour developing of iodine, record Rf (initial point is to the distance of spot centers and the ratio of initial point to the distance of solvent front) value different with FFC, prove that FFA generates.By solution left at room temperature over night, with Filter paper filtering, distilled water washing to the FFA haptens after trickle is colourless, drying, synthetic route is as follows:
Embodiment 2.
The preparation of comlete antigen: take 25mg florfenicol amine and be dissolved in 2mLDMF; Weighing 66mgBSA is dissolved in the PBS damping fluid of 10mlPH7.4; Florfenicol amine is dropwise joined in above-mentioned BSA solution, stir under 4 DEG C of conditions, then dropwise add 100 μ L glutaraldehydes in reactant liquor, slow stirring reaction 24h.Dialyse the product PBS of synthesis 72h, every 8h changes liquid once, obtain florfenicol amine-bovine serum albumin(BSA) (FFA-BSA) immunogene, with legal system for florfenicol amine-oralbumin (FFA-OVA) coating antigen, synthetic route is as follows:
Embodiment 3.
The preparation of the monoclonal antibody of anti-florfenicol amine: the hybridoma of the monoclonal antibody of the anti-florfenicol amine of secretion of the present invention, its preparation is as follows with the method selected:
(1) alkali hydrolysis method prepares florfenicol amine (FFA) haptens
(2) with glutaraldehyde method synthesis florfenicol amine comlete antigen, immunogene FFA-BSA is comprised, coating antigen FFA-OVA;
(3) with immunogene FFA-BSA immunity Balb/c small white mouse;
(4) by the splenocyte of immune mouse and the fusion of myeloma cell, 120 strain of hybridoma are cultivated to obtain;
(5) with the strain of coating antigen FFA-OVA wrapper sheet indirect elisa method screening positive hybridoma cell;
(6) limiting dilution assay is adopted to clone wherein 4 strain positive hybridoma cells;
(7) antibody prepared by the cell line being filtered out monoclonal antibody 2D6-1C4 bis-time cloning of the highest anti-florfenicol amine of tiring by indirect elisa method, preparation gold mark monoclonal antibody.In table 1
Table 1: monoclonal antibody the selection result
Embodiment 4.
The specificity identification of florfenicol amine monoclonal antibody C of the present invention: detect with anti-florfenicol amine monoclonal antibody microbiotic different from the 4 kinds ELLIA that is at war with, use FFA-OVA wrapper sheet.Negative control PBS wrapper sheet; Sun contrasts, and replaces the microbiotic for detecting cross reaction with PBS.Testing result, florfenicol amine monoclonal antibody can be reacted with florfenicol amine and also can be reacted with Florfenicol, both indifferences.And florfenicol amine monoclonal antibody and similar microbiotic Thiamphenicol and other classes are as the terramycin of macrolides, the norfloxacin hydrochloride of quinolones and the equal no cross reaction of the radonil of sulfamido.This antibody high specificity is described, and the former medicine of Florfenicol can not only be detected also can detects metabolic product florfenicol amine in its body.In table 2
Table 2: anti-florfenicol amine monoclonal antibody specificity experiment (
n=6)
Note: *. compare with positive controls, OD value significantly reduces, (P < 0.01)
Embodiment 5
Florfenicol Rapid detection test strip, see accompanying drawing 1, it comprises: the anti-florfenicol amine of preserving number secreted by CCTCC-C201575 hybridoma FFA-C, the monoclonal antibody C (being called for short: gold mark monoclonal antibody C) of colloid gold label, anti-florfenicol amine polyclonal antibody (being called for short: resist) and goat anti-mouse igg (being called for short: sheep anti-mouse igg) more, with the carrier board 3 of adhesive sticker, at the both ends of carrier board 3, be respectively equipped with application of sample end water accepting layer 1 and handheld terminal water accepting layer 7, the detection layers 6 be made up of nitrocellulose membrane is provided with at this carrier board 3 middle part, be provided with at this detection layers 6 and application of sample end water accepting layer 1 intersection and be loaded with the glass layer block 2 that gold marks monoclonal antibody C, one end of glass layer block 2 is arranged under application of sample end water accepting layer 1, the other end is arranged on detection layers 6, the detection layers 6 continued with glass layer block 2 is provided with detection line 4 and nature controlling line 5, detection line 4 and nature controlling line 5 place are coated with many anti-and sheep anti-mouse iggs respectively.
Embodiment 6
The preparation process of the monoclonal antibody C of colloid gold label is as follows:
(1) preparation of collaurum: prepared by 10-20nm colloid gold particle, be that 0.01% chlorauric acid solution 250ml mixes with 1% sodium citrate 6.65ml by concentration, be heated to 100 degrees Celsius, make colloidal gold solution, the pH value of this solution: be adjusted to 8.2 with 0.2M sal tartari, for subsequent use;
(2) preparation of gold mark monoclonal antibody C:
When anti-florfenicol amine monoclonal antibody concentration is 1g/L, the suitableeest monoclonal antibody amount of 100ml colloid gold label is 200 μ l, and join in colloidal gold solution in this ratio by anti-florfenicol amine monoclonal antibody amine, stir 50min slowly, add 5%PEG20000,4 DEG C are spent the night; Then by it at 4 DEG C, the centrifugal 60min of 8000r; Get centrifugation, with the 0.01mol/L phosphate buffer (KCL0.2g contained containing 5%PEG20000,0.25% polysorbas20, NaCL8.0g, KH2PO40.2g, Na2HPO412H2O2.9g, distilled water 1000ml, pH7.4) hang, add Sodium azide again and concentration is adjusted to 0.02%, namely make gold mark monoclonal antibody C.
Embodiment 7.
The preparation that how anti-anti-florfenicol amine is: with the comlete antigen (FFA-OVA) of florfenicol amine and ovalbumin coupling for immunogen immune rabbit, be coating antigen with the comlete antigen (FFA-BSA) of florfenicol amine and bovine serum albumin(BSA) coupling, carry out immune enzyme-linked sorbent assay (ELISA), recording tires when reaching 1:100000, Culling heart blood separation of serum, 4 DEG C save backup.
Embodiment 8.
Florfenicol Site Detection detection paper layer preparation method of the present invention is: purify the rabbit anteserum of immunity by saturated ammonium sulfate method, after freeze drying, be prepared into the anti-florfenicol amine of 1mg/ml to resist more, be sprayed on nitrocellulose membrane with Membrane jetter, form detection line 4; Equally, by goat anti-mouse igg, be prepared into 200 μ g/ml, be sprayed on nitrocellulose membrane with Membrane jetter, form nature controlling line 5.Two lines are at a distance of 5mm, and the nearly handheld terminal water accepting layer of nature controlling line 5, detection line 4 is application of sample end water accepting layer closely.Dry under room temperature, then use pH7.4,0.010.01mol/LPBS37 DEG C of closed 30min, the PBS rinsing containing 5%PEG20000 is dried.
Embodiment 9.
The minimum residual quantity that the quick detector bar of Florfenicol of the present invention detects is 10ug/ml.In table 3
Table 3: the mensuration that Florfenicol Site Detection test paper is sensitive
Note: ++ represent that detection line color is aubergine, testing result is strong positive; + representing that detection line color is red, testing result is positive;-representing that detection line is colourless, testing result is negative.
Embodiment 10.
The using method of Florfenicol Rapid detection test strip of the present invention, detecting step is as follows: first, live fish one is got from the pond of cultivation, get liver, by weight/volume (w/v) adds the phosphate buffer of physiological saline or pH6.0-8.0 than 1:2, liver is pulverized, is prepared into detection sample; Then, immersed by application of sample end 1 water accepting layer of test paper in the detection sample of test tube (liquid level must not exceed 2/3 of thieving paper 1), 5min reads result.
Positive findings: inspected layer about 6 presents two red detection lines 4 and nature controlling line 5, represents that sample residually has Florfenicol or florfenicol amine Florfenicol in having;
Negative findings: inspected layer 6 upper end presents a red nature controlling line 5, represents that sample represents and can't detect Florfenicol, florfenicol amine or both concentration lower than 10ug/ml;
Testing result is invalid: in application of sample 5 minutes, nature controlling line 5 place, inspected layer 6 upper end occurs without red line, and represent that this test paper lost efficacy, testing result is invalid.(see accompanying drawing 2)
Claims (7)
1. Florfenicol Site Detection test paper, it is characterized in that comprising: the anti-florfenicol amine of preserving number secreted by CCTCC-C201575 hybridoma FFA-C, the monoclonal antibody C (being called for short gold mark monoclonal antibody C) of colloid gold label, anti-florfenicol amine polyclonal antibody and goat anti-mouse igg, with the carrier board (3) of adhesive sticker, at the both ends of carrier board (3), be respectively equipped with application of sample end water accepting layer (1) and handheld terminal water accepting layer (7), the detection layers (6) be made up of nitrocellulose membrane is provided with at this carrier board (3) middle part, be provided with at this detection layers (6) and application of sample end water accepting layer (1) intersection and be loaded with the glass layer block (2) that gold marks monoclonal antibody C, one end of glass layer block (2) is arranged under application of sample end water accepting layer (1), the other end is arranged on detection layers (6), the detection layers (6) continued with glass layer block (2) is provided with detection line (4) and nature controlling line (5), detection line (4) and nature controlling line (5) place are coated with anti-florfenicol amine polyclonal antibody and goat anti-mouse igg respectively.
2. Florfenicol Site Detection test paper according to claim 1, it is characterized in that described gold mark monoclonal antibody C prepares by alkali hydrolysis method the anti-florfenicol amine monoclonal antibody that florfenicol amine (FFA) haptens and bovine serum albumin(BSA) (BSA) prepared as immunogene by glutaraldehyde method coupling complete antigen synthesis FFA-BSA, prepare after colloid gold label.
3. Florfenicol Site Detection test paper according to claim 1, is characterized in that described anti-florfenicol amine polyclonal antibody prepares by alkali hydrolysis method the rabbit anteserum that florfenicol amine (FFA) haptens and oralbumin (OVA) prepared as immunogen immune rabbit by glutaraldehyde method coupling complete antigen synthesis (FFA-OVA).
4. Florfenicol Site Detection test paper according to claim 1, is characterized in that the preparation method of described gold mark monoclonal antibody C is as follows:
(1) prepare colloidal gold solution with the preparation technology of known collaurum, the grain size of obtained collaurum is 10-20nm;
(2) by monoclonal antibody C (0.5-1g/L antibody protein), join in above-mentioned 100ml colloidal gold solution by 150-500 μ l, mix even slowly;
(3) in mixed solution, add PEG20000, make its final concentration be 1-5%, hold over night;
(4) centrifugal 8000-10000 rev/min, 1-1.5 hour, abandon supernatant;
(5) get centrifugation, hang with the 0.01mol/L phosphate buffer containing 1-5%PEG20000,0.05-0.25% Tween-20,0.02% sodium azide, make gold mark monoclonal antibody C.
5., according to Florfenicol Site Detection test paper according to claim 4, it is characterized in that gold chloride mixes with trisodium citrate and heat by certain proportioning to be prepared into colloidal gold solution by the preparation method of colloidal gold solution described in step (1).
6., according to Florfenicol Site Detection test paper according to claim 4, it is characterized in that in step (2), monoclonal antibody C and colloidal gold solution mix even 30-60min slowly.
7. the using method of Florfenicol Site Detection test paper, it is characterized in that comprising the following steps: the application of sample end water accepting layer (1) of this test paper is inserted or drips detected sample thereon, 5-10min is being positioned at the detection line (4) of detection layers (6) lower end and is being positioned at nature controlling line (5) the place display redness of detection layers (6) upper end, i.e. two red line, representing residual in sample has Florfenicol or florfenicol amine; Detection line place is without red line, and represent that sample represents and can't detect Florfenicol, florfenicol amine or both concentration lower than 10ug/ml, the display of nature controlling line place is red, represents that this test paper is effective; Nature controlling line place does not show redness, represents that this test paper is invalid.
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Cited By (5)
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| CN106771159A (en) * | 2016-12-01 | 2017-05-31 | 无锡艾科瑞思产品设计与研究有限公司 | A kind of Florfenicol Rapid detection test strip |
| CN106905428A (en) * | 2017-04-25 | 2017-06-30 | 江南大学 | A kind of synthetic method of florfenicol amine artificial antigen |
| CN109709257A (en) * | 2019-01-30 | 2019-05-03 | 武汉生物工程学院 | A kind of method of Florfenicol and metabolite florfenicol amine in detection pig urcine |
| CN109709258A (en) * | 2019-01-30 | 2019-05-03 | 武汉生物工程学院 | A kind of method and application detecting Florfenicol total residual object in pig edible tissue |
| CN110988350A (en) * | 2019-11-08 | 2020-04-10 | 南京市产品质量监督检验院 | Quantum dot test paper capable of quantitatively detecting florfenicol residual quantity in various matrixes |
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| CN109709257A (en) * | 2019-01-30 | 2019-05-03 | 武汉生物工程学院 | A kind of method of Florfenicol and metabolite florfenicol amine in detection pig urcine |
| CN109709258A (en) * | 2019-01-30 | 2019-05-03 | 武汉生物工程学院 | A kind of method and application detecting Florfenicol total residual object in pig edible tissue |
| CN109709258B (en) * | 2019-01-30 | 2021-07-20 | 武汉生物工程学院 | Method for detecting florfenicol total residues in edible tissues of pigs and application |
| CN110988350A (en) * | 2019-11-08 | 2020-04-10 | 南京市产品质量监督检验院 | Quantum dot test paper capable of quantitatively detecting florfenicol residual quantity in various matrixes |
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