CN203455348U - Immune colloidal gold combined reagent plate for detecting clenbuterol hydrochloride, ractopamine and salbutamol in swine urine - Google Patents
Immune colloidal gold combined reagent plate for detecting clenbuterol hydrochloride, ractopamine and salbutamol in swine urine Download PDFInfo
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- CN203455348U CN203455348U CN201320566244.7U CN201320566244U CN203455348U CN 203455348 U CN203455348 U CN 203455348U CN 201320566244 U CN201320566244 U CN 201320566244U CN 203455348 U CN203455348 U CN 203455348U
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Abstract
The utility model relates to an immune colloidal gold combined reagent plate for detecting clenbuterol hydrochloride, ractopamine and salbutamol in swine urine. The reagent plate is composed of an upper plastic mold plate, a lower plastic mold plate, a clenbuterol hydrochloride rapid detection test strip, a ractopamine rapid detection test strip and a salbutamol rapid detection test strip, wherein each test strip consists of a bottom plate, as well as a sample pad, a gold-labeled combined pad, a nitrocellulose membrane and an absorbent pad which are arranged on the bottom plate sequentially, and a detection line and a quality control line are sequentially sprayed on the nitrocellulose membrane of each test strip in a direction from the sample pad to the absorbent pad. The reagent plate can be used for intuitive semi-quantitative detection, the whole operation process is about 5 minutes, no expensive laboratory equipment is required for assisting, large-scale sample selection is facilitated, and the immune colloidal gold combined reagent plate is suitable for slaughter enterprises and government detection institutions.
Description
Technical field
The utility model relates to the immune colloid gold associating agent plate of clenobuterol hydrochloride, Ractopamine, salbutamol in a kind of urine of fast detecting pig simultaneously.
Background technology
Clenobuterol hydrochloride (Clenbuterol, CLEN), salbutamol (Salbutamol, SAL), Ractopamine (Ractopamine, RAC), belong to beta-stimulants, is commonly called " clenbuterol hydrochloride ".Clenobuterol hydrochloride can be used for preventing and treating the pulmonary disease such as asthma, pulmonary emphysema clinically, and salbutamol can be used for treating bronchial astehma, and Ractopamine can be used for treating the diseases such as congestive heart failure disease.Research finds to use for a long time or in a large number beta-stimulants can make the synthetic increase of animal muscle, and fat deposition reduces, and obviously promotes growth of animal, and therefore, it is often used as feed addictive.Because beta-stimulants metabolism is in animal body slow, long-time use can cause accumulation, when having eaten, the mankind contain after the residual consumption animal of beta-stimulants, there will be toxicity symptom in various degree, common have nauseating, dizzy, weakness of limbs, a hand tremor etc., and heart disease, hyperpietic's harm are especially serious.
For this reason, European Union clearly forbids using in consumption animal all beta-stimulants that comprise clenobuterol hydrochloride, salbutamol, Ractopamine etc.For ensureing the healthy of the sustainable and healthy development of animal husbandry and the people, clenobuterol hydrochloride, salbutamol, Ractopamine have also been listed in the forbidding inventory such as in the types of drugs catalogue > > that No. 176 bulletin < < forbid using in feed and animal drinking water by China Ministry of Agriculture.Although China prohibites use, yet, driven by economic interests and lack effective monitoring means, still have at present many raisers use beta-stimulants of disobeying orders, therefore, the medicament residue method for supervising of setting up science, standard is very necessary.
Present stage, for the detection method of beta-stimulants, mainly contain physico-chemical analysis method and immunoassay.Physico-chemical analysis method, as high performance liquid chromatography (HPLC) and liquid chromatography mass coupling method (LC-MS), these methods have the advantages that specificity is good, accuracy rate is high, the prefered method that Shi Ge great testing agency confirms detecting sample, but equipment, environment, operative skill etc. are had relatively high expectations, be unfavorable for extensive screening sample, thereby be not suitable for department of vast basic unit.Immunoassay is conventional enzyme linked immunosorbent assay (ELISA) and colloidal gold immunity chromatography (GICA).
Enzyme linked immunosorbent assay is the immune analysis method that the efficient catalytic of application antigen and antibody specific reaction and enzyme is used for measuring beta-stimulants content, has high specificity, the advantage such as highly sensitive, quick, easy.But in ELISA method, the activity of enzyme is subject to reaction conditions impact, causes measurement result repeatability poor; And need to use expensive special instrument and equipment, therefore have some limitations.
Colloidal gold immunity chromatography is the immunology detection technology of a kind of simple and fast of setting up on the basis of immunity percolation technology.The method is incorporated into the needed whole or most of raw materials of reaction in reagent, can be in 3～5min the mensuration of complete paired samples, have simple, fast, the feature such as sensitivity is high, both can be used for laboratory to detect, also can be used for plant etc. and measure on the spot.
Utility model content
The utility model detects demand for reality, develops the immune colloid gold associating agent plate of clenobuterol hydrochloride, Ractopamine, salbutamol in a kind of urine of fast detecting pig simultaneously.The utility model agent plate is simple and efficient to handle, highly sensitive, and specificity is good, and experimental result naked eyes can intuitive judgment, without being equipped with instrument and equipment, can complete the mensuration to above-mentioned three kinds of medicament residues in pig urine in 3～5min.
The utility model agent plate is comprised of upper and lower two plastic formworks and fast clenbuterol hydrochloride detecting test paper strip, Ractopamine Rapid detection test strip, salbutamol Rapid detection test strip.Every test strips forms by the sample pad of sticking together successively on base plate and base plate, gold-marking binding pad, nitrocellulose filter and adsorptive pads, and adjacent part has the overlapping region of 1～2mm.Sample pad, after the immersion of salt ion damping fluid and activating agent is dry, is attached with the materials such as salt ion and activating agent above.On gold-marking binding pad, be coated with respectively clenobuterol hydrochloride monoclonal antibody-colloid gold label thing, Ractopamine monoclonal antibody-colloid gold label thing and salbutamol monoclonal antibody-colloid gold label thing.Nitrocellulose filter is sprayed with detection line and nature controlling line from sample pad successively to adsorptive pads direction, on the detection line of three kinds of test strips, be coated with respectively clenobuterol hydrochloride-carrier protein couplet thing, Ractopamine-carrier protein couplet thing, salbutamol-carrier protein couplet thing, on nature controlling line, be all coated with sheep anti-mouse igg antibody.The carrier protein of coupling clenobuterol hydrochloride, Ractopamine, salbutamol can be ovalbumin (OVA), bovine serum albumin(BSA) (BSA), hemocyanin (KLH) etc.
In the utility model agent plate, the reaction principle of contained fast clenbuterol hydrochloride detecting test paper strip, Ractopamine Rapid detection test strip, salbutamol Rapid detection test strip is identical.For example fast clenbuterol hydrochloride detecting test paper strip has used colloidal gold-labeled method, chromatographic technique, competition immunological technique, by antigen and monoclonal antibody-colloid gold label thing reaction solution, and the clenobuterol hydrochloride medicament residue level in specific detection pig urine.If contain clenobuterol hydrochloride residue in sample solution, the clenobuterol hydrochloride monoclonal antibody reaction of clenobuterol hydrochloride first and on colloid gold particle, when colloid gold particle diffuses to detection line with sample solution, on colloid gold particle, the avtive spot of clenobuterol hydrochloride monoclonal antibody cannot be combined by clenobuterol hydrochloride-carrier protein couplet thing because the clenobuterol hydrochloride medicine by sample solution occupies on detection line; When the clenobuterol hydrochloride content in sample surpasses agent plate detection limit, the detection line in agent plate, without colour developing, is judged to be the positive.Otherwise clenobuterol hydrochloride content is below agent plate detection limit or during noresidue in sample, the detection line colour developing in agent plate, is judged to be feminine gender.
Each several part composition and the function of the utility model agent plate are as follows:
Plastic formwork, plays fixed base plate and indicates each functional areas (well, detection zone, Quality Control district).
Base plate, is made by Polyvinylchloride (PVC) material that simultaneously scribbles adhesive sticker, plays other ingredients of fixed support agent plate.
Sample pad, is made by glass fibre, works to absorb sample solution and buffering sample solution pH value.
Gold-marking binding pad, is made by glass fibre, for the reaction of effective constituent in sample solution and monoclonal antibody-colloid gold label thing provides place.
Nitrocellulose filter, is sprayed with detection line and nature controlling line from sample pad to adsorptive pads direction, by reaction result with macroscopic characterization out successively.
Adsorptive pads, is made by filter paper, unnecessary solution in absorption reaction process.
The utility model agent plate has following beneficial effect:
(1) specificity is good.In the utility model agent plate, fast clenbuterol hydrochloride detecting test paper strip is 100% to the cross reacting rate of clenobuterol hydrochloride, to if the cross reacting rate of other beta-stimulants such as salbutamol, Ractopamine is all lower than 1%; Ractopamine Rapid detection test strip is 100% to the cross reacting rate of Ractopamine, to if the cross reacting rate of other beta-stimulants such as salbutamol, clenobuterol hydrochloride is all lower than 1%; Salbutamol Rapid detection test strip is 100% to the cross reacting rate of salbutamol, to the cross reacting rate of other beta-stimulants such as example hydrochloric acid Clenbuterol, Ractopamine all lower than 1%.Visible, the utility model agent plate respectively has height selectivity to three kinds of residual detections.
(2) highly sensitive.The utility model agent plate is 3.0ppb for the detection limit of clenobuterol hydrochloride, Ractopamine, salbutamol in pig urine samples, is applicable to all kinds of enterprises and testing agency.
(3) simple and quick.The utility model agent plate is incorporated into the required most of raw material of reaction in test strips, after a sample, reacts and carries out fast on immobilon-p, gets final product reading result in 3～5min, can detect three kinds of medicament residues simultaneously, has greatly shortened the sample time.The use of this agent plate does not need any professional training, and ordinary person all can operate.
(4) do not rely on experimental facilities.The utility model agent plate is after directly dripping sample race plate, by naked eyes, judge that detection line on nitrocellulose filter and the shade of nature controlling line carry out sentence read result, whole testing process does not rely on any experimental facilities, is particularly suitable for field and execute-in-place, is easy to promote.
(5) cost is low, easily promotes.The utility model agent plate production technology is simple, can realize single pattern detection, and production cost is low, greatly reduces testing cost.
Accompanying drawing explanation
Fig. 1 is the side structure schematic diagram of test strips in the utility model agent plate.Wherein 1 is sample pad, and 2 is gold-marking binding pad, and 3 is nitrocellulose filter (NC film), and 4 is detection line (T line), and 5 is nature controlling line (C line), and 6 is adsorptive pads, and 7 is adhesive sticker, and 8 is polyvinyl chloride panel (PVC plate).
Fig. 2 is the utility model agent plate operation chart, and wherein S is well, and C is Quality Control district, and T is detection zone, and 1 represents clenobuterol hydrochloride, and 2 represent Ractopamine, and 3 represent salbutamol.
Fig. 3 is that the utility model agent plate result is judged schematic diagram, and wherein C is Quality Control district, and T is detection zone.
Specific implementation method
The preparation of the utility model agent plate comprises the preparation of hapten-carrier protein conjugate, the preparation of monoclonal antibody, and the preparation of colloidal gold solution, the preparation of monoclonal antibody-colloid gold label thing, the assembling of agent plate, detects implementation method.
1. the preparation of hapten-carrier protein conjugate
The preparation of 1.1 clenobuterol hydrochlorides-carrier protein couplet thing
Take 20mg clenobuterol hydrochloride, with the sulfuric acid solution of 2mL0.5mol/L, dissolve, after mixing, add the sodium nitrite of 300 μ L10mg/mL, sustained response 2h.Under agitation, solution is dropwise joined to (100mg BSA is dissolved in the sodium carbonate buffer solution of 10mL0.1mol/LpH10.0) in BSA solution.After mixing, be placed at 4 ℃ lucifuge reaction 5h (during can put upside down mix), afterwards with the PBS damping fluid dialysis of 0.01mol/LpH7.4 4 days, during at interval of 8h, change liquid once.Again by solution centrifugal, collect supernatant, frozen standby at-20 ℃.
With ovalbumin (OVA), substitute BSA, adopt same method to prepare clenobuterol hydrochloride-ovalbumin conjugate.
The preparation of 1.2 Ractopamines-carrier protein couplet thing
Take 34mg hydrochloric acid Ractopamine, be dissolved in 2mL pyridine, add 12mg glutaric anhydride, under room temperature, stir and spend the night, nitrogen dries up; Product is dissolved into 4mL N ' dinethylformamide (DMF) and 1, in 4-dioxane mixed solution (both volume ratios are 1: 1), add 26.2 μ L tri-n-butylamines, ice bath stirs 10min, add isobutyl chlorocarbonate 14.3 μ L, stirring at room reaction 1h; Said mixture is dropwise joined in the BSA solution of precooling (100mg BSA is dissolved in the dobell's solution of 0.1mol/LpH8.5) to stirred overnight at room temperature; With the PBS damping fluid of 0.01moL/L pH7.4, dialyse 3 days, change liquid every day 3 times.
With ovalbumin (OVA), substitute BSA, adopt same method to prepare Ractopamine-ovalbumin conjugate.
The preparation of 1.3 salbutamols-carrier protein couplet thing
Prepare salbutamol alkali: claim 2.4g salbutamol sulfate, with 30mL absolute methanol, dissolve, cross anion exchange resins, collect eluent, recrystallization, obtains white crystal (fusing point 154-155 ℃).
Synthetic salbutamol hemisuccinic acid: claim 240mg salbutamol alkali, dissolve with 20mL methyl alcohol; Rotary evaporation is yellow oil, then uses anhydrous alcohol solution, the 120mg succinic anhydride that dropwise adds ethanol to dissolve; Under nitrogen protection condition, stirring at room 4h; Reactant liquor is centrifugal, abandon supernatant, use absolute ethanol washing sediment 3 times, volatilizing solvent gained white solid is haptens.
Mixed anhydride method coupling salbutamol and carrier protein: claim 40mg half succinyl salbutamol, be dissolved in the potpourri of Isosorbide-5-Nitrae-dioxane-water-triethylamine, under stirring, dropwise add 30 μ L isobutyl chlorocarbonates (in 30min); Above-mentioned reactant liquor is dropwise joined to 25mL containing in the distilled water of 80mg BSA; 4 ℃ of reactions are spent the night, and the PBS damping fluid dialysis of 0.01moL/L pH7.4 3 days, changes liquid every day 3 times.
With ovalbumin (OVA), substitute BSA, adopt same method to prepare salbutamol-ovalbumin conjugate.
2. the preparation of monoclonal antibody
The preparation of 2.1 clenobuterol hydrochloride monoclonal antibodies
Get female Balb/C mouse in 6～8 week age, will be as immunogenic clenobuterol hydrochloride-bovine serum albumin(BSA) conjugate (CLEN-BSA) and isopyknic Freund's complete adjuvant emulsification, by a 100 μ g/ dosage hypodermic injection, afterwards every 3 weeks booster immunizations 1 time, the full adjuvant that toos many or too much for use replaces Freund's complete adjuvant and antigen to mix carrying out lumbar injection.Merge first 3 days reinforced immunologicals 1 time, without adjuvant, dosage doubles.Fusion of Cells is carried out according to a conventional method: Sp2/0 multiple myeloma cells is mixed in the ratio of 1: 10 with immune spleen cell, under 50%PEG effect, merge, HAT nutrient culture media suspends, and divides and plants in 96 well culture plates, 37 ℃, 5%CO
2in incubator, cultivate.
After fusion, until Growth of Cells, arrive 1/4 o'clock of culture hole area, adopt a minute step screening method screening hybridoma.Primary dcreening operation is selected the coated elisa plate of 10mg/L clenobuterol hydrochloride-ovalbumin conjugate (CLEN-OVA), and measured hole adds culture supernatant, after hatching, cleaning, adds sheep anti-mouse igg-HRP (1: 1000), OPD colour developing.The coated elisa plate of positive Kong Zaiyong CLEN-OVA filtering out is blocked indirect ELISA.By cells and supernatant and 2 * 10
-3mol/L clenobuterol hydrochloride solution mixed in equal amounts, 1h is made in 37 ℃ of senses, adds in coated ELISA Plate.Use in addition PBS (0.01mol/L, pH7.4) Instead of Hydrochloric Clenbuterol solution to compare, all the other steps are the same.If the OD value after clenobuterol hydrochloride blocking-up is down to below 50% of control wells, be judged to positive hole.The hole of being all positive through 2～3 detections, carries out cloning with limiting dilution assay immediately.
In vitro culture: the cell line of cloning is expanded and cultivated, and cell concentration reaches 5 * 10
5during/mL, stop changing liquid, cell is the dead rear nutrient solution of collecting all.
In body, induce ascites: the mouse peritoneal injection cloning cell line 10 to lumbar injection whiteruss after 10 days
7individual cell, extracted ascites after 7 days.
The preparation of 2.2 Ractopamines, salbutamol monoclonal antibody
Adopt method of the same race (step 2.1) to prepare Ractopamine monoclonal antibody, salbutamol monoclonal antibody.
3. the preparation of colloidal gold solution
The mean size of colloid gold particle is 30nm, and its preparation method, for to add 1mL1% trisodium citrate in 100mL deionized water, boils the rear 1mL1% gold chloride that adds rapidly, continues to boil 10min, cooling after, save backup at 4 ℃.
4. the preparation of monoclonal antibody-colloid gold label thing
The preparation of 4.1 clenobuterol hydrochloride monoclonal antibody-colloid gold label things
Before mark by monoclonal antibody solution with 15000r/min4 ℃ of centrifugal 30min, get supernatant; Get the colloidal gold solution 100mL having prepared, use 0.1mol/L K
2cO
3with 0.1mol/L HCl, the pH value of colloidal gold solution is adjusted to slightly meta-alkali (pH=pI+0.5) of monoclonal antibody isoelectric point; Add while stirring 0.66mg clenobuterol hydrochloride monoclonal antibody; Under agitation dropwise add 2mL25mol/L PEG 20000 (PEG20000), stir 15min; The centrifugal 15min of 20000r/min, abandons supernatant; The PBS damping fluid (containing 0.4mol/L PEG20000) that adds 10mL0.01mol/LpH7.4, the centrifugal 15min of 20000r/min, abandons supernatant, so washs 2～4 times, thoroughly to remove unconjugated protein; Precipitation is dissolved containing the PBS damping fluid (pH7.4,0.01mol/L) of 2%BSA with 10mL; After filtering with 0.22 μ m sterilizing filter, obtain clenobuterol hydrochloride monoclonal antibody-colloid gold label thing, 4 ℃ save backup.
The preparation of 4.2 Ractopamine monoclonal antibody-colloid gold label things
With 2mg Ractopamine monoclonal antibody, substitute 0.66mg clenobuterol hydrochloride monoclonal antibody, same method (step 4.1) is prepared Ractopamine monoclonal antibody-colloid gold label thing.
The preparation of 4.3 salbutamol monoclonal antibody-colloid gold label things
With 1.5mg salbutamol monoclonal antibody, substitute 0.66mg clenobuterol hydrochloride monoclonal antibody, same method (step 4.1) is prepared salbutamol monoclonal antibody-colloid gold label thing.
5. the assembling of agent plate
With reference to Fig. 1, with some film machine, clenobuterol hydrochloride-carrier protein couplet thing of debita spissitudo and sheep anti-mouse igg antibody are sprayed on nitrocellulose filter, respectively as detection line and nature controlling line, 37 ℃ of oven drying 8h; Clenobuterol hydrochloride monoclonal antibody-colloid gold label the thing preparing is coated on gold-marking binding pad; Sample pad, gold-marking binding pad, nitrocellulose filter and adsorptive pads are sticked on PVC base plate successively; With cutting machine, the kilocalorie posting is cut into the wide test strips of 3mm, complete the preparation of fast clenbuterol hydrochloride detecting test paper strip.
With legal system for Ractopamine Rapid detection test strip, salbutamol Rapid detection test strip.
Above-mentioned three kinds of test strips are packed in plastic formwork after generate a reagent plate, then put into the aluminium foil bag sealed storage with drying agent.
6. detect implementation method
6.1 sample preparation
By dry, clean centrifuge tube or appropriate containers, gather 5mL left and right pig urine, directly as detecting sample, if precipitation or muddy thing appear in urine sample, get supernatant as detecting sample after centrifugal.
6.2 detecting step
From packaging bag, take out agent plate, draw sample solution to be checked and be added drop-wise to respectively in three wells, every hole 100 μ L, start timing after application of sample; Result should read at 3～5min, and other times interpretation is invalid.
6.3 result interpretations
During reading result, agent plate level is placed in to observer front, as shown in Fig. 2 right side.
The interpretation of clenobuterol hydrochloride residue:
Negative (-): detection line (T line) colour developing on clenobuterol hydrochloride examination fast detecting paper slip, represents in sample that clenobuterol hydrochloride content is lower than 3.0ppb or not hydrochloric Clenbuterol.As shown in Fig. 3 .a.
Positive (+): the detection line on fast clenbuterol hydrochloride detecting test paper strip (T line), without colour developing, represents in sample that clenobuterol hydrochloride content is greater than or equal to 3.0ppb.As shown in Fig. 3 .b.
Invalid: not occurring nature controlling line (C line), may be that misoperation or agent plate lost efficacy.Should again read instructions, and retest by new agent plate.As shown in Fig. 3 .c.
With method interpretation Ractopamine, salbutamolum residue.
Claims (6)
1. in an energy while fast detecting pig urine, the immune colloid gold of clenobuterol hydrochloride, Ractopamine, salbutamol is combined agent plate, it is characterized in that agent plate is comprised of upper and lower two plastic formworks and fast clenbuterol hydrochloride detecting test paper strip, Ractopamine Rapid detection test strip, salbutamol Rapid detection test strip, every test strips forms by the sample pad of sticking together successively on base plate and base plate, gold-marking binding pad, nitrocellulose filter and adsorptive pads, and adjacent part has the overlapping region of 1～2mm.
2. agent plate as claimed in claim 1, the nitrocellulose filter that it is characterized in that clenobuterol hydrochloride, Ractopamine, salbutamol Rapid detection test strip is sprayed with detection line and nature controlling line from sample pad successively to adsorptive pads direction, on detection line, be coated with respectively clenobuterol hydrochloride-carrier protein couplet thing, Ractopamine-carrier protein couplet thing, salbutamol-carrier protein couplet thing, on nature controlling line, be all coated with sheep anti-mouse igg antibody.
3. agent plate as claimed in claim 1, is characterized in that the gold-marking binding pad of clenobuterol hydrochloride, Ractopamine, salbutamol Rapid detection test strip is coated with respectively clenobuterol hydrochloride monoclonal antibody-colloid gold label thing, Ractopamine monoclonal antibody-colloid gold label thing, salbutamol monoclonal antibody-colloid gold label thing.
4. agent plate as claimed in claim 1, the mean size that it is characterized in that colloid gold particle is 30nm.
5. agent plate as claimed in claim 1, is characterized in that the carrier protein of coupling Ractopamine, clenobuterol hydrochloride and salbutamol can be bovine serum albumin(BSA) (BSA), ovalbumin (OVA), hemocyanin (KLH).
6. agent plate as claimed in claim 1, is characterized in that base plate made by the pvc material that simultaneously scribbles adhesive sticker, and sample pad is made by glass fibre, and gold-marking binding pad is made by glass fibre, and adsorptive pads is filter paper.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201320566244.7U CN203455348U (en) | 2013-09-09 | 2013-09-09 | Immune colloidal gold combined reagent plate for detecting clenbuterol hydrochloride, ractopamine and salbutamol in swine urine |
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| CN201320566244.7U CN203455348U (en) | 2013-09-09 | 2013-09-09 | Immune colloidal gold combined reagent plate for detecting clenbuterol hydrochloride, ractopamine and salbutamol in swine urine |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104280540A (en) * | 2014-10-21 | 2015-01-14 | 南京师范大学 | Immunosensor and method for detecting various beta-adrenergic receptor stimulant residues |
| CN104280541A (en) * | 2014-10-21 | 2015-01-14 | 南京师范大学 | Beta-epinephrine receptor stimulant multi-cluster antigens and wide-spectrum specific antibodies and preparation thereof |
| CN106896094A (en) * | 2017-04-11 | 2017-06-27 | 中国农业科学院饲料研究所 | It is a kind of at the same detect CLE, RAC and SBL method and its special paper chip |
| CN112903875A (en) * | 2021-01-25 | 2021-06-04 | 中国农业科学院农产品加工研究所 | Ractopamine standard substance with conjugated state and free state coexisting in pig urine matrix as well as preparation method and application thereof |
| CN112903874A (en) * | 2021-01-25 | 2021-06-04 | 中国农业科学院农产品加工研究所 | Free and conjugated clenbuterol standard substance contained in pig urine and preparation method thereof |
| CN112903873A (en) * | 2021-01-25 | 2021-06-04 | 中国农业科学院农产品加工研究所 | Free-state and conjugated-state salbutamol standard substance contained in swine urine freeze-dried powder after animal metabolism and preparation method thereof |
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2013
- 2013-09-09 CN CN201320566244.7U patent/CN203455348U/en not_active Expired - Lifetime
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104280540A (en) * | 2014-10-21 | 2015-01-14 | 南京师范大学 | Immunosensor and method for detecting various beta-adrenergic receptor stimulant residues |
| CN104280541A (en) * | 2014-10-21 | 2015-01-14 | 南京师范大学 | Beta-epinephrine receptor stimulant multi-cluster antigens and wide-spectrum specific antibodies and preparation thereof |
| CN106896094A (en) * | 2017-04-11 | 2017-06-27 | 中国农业科学院饲料研究所 | It is a kind of at the same detect CLE, RAC and SBL method and its special paper chip |
| CN106896094B (en) * | 2017-04-11 | 2023-07-14 | 中国农业科学院饲料研究所 | Method for simultaneously detecting CLE, RAC and SBL and special paper chip thereof |
| CN112903875A (en) * | 2021-01-25 | 2021-06-04 | 中国农业科学院农产品加工研究所 | Ractopamine standard substance with conjugated state and free state coexisting in pig urine matrix as well as preparation method and application thereof |
| CN112903874A (en) * | 2021-01-25 | 2021-06-04 | 中国农业科学院农产品加工研究所 | Free and conjugated clenbuterol standard substance contained in pig urine and preparation method thereof |
| CN112903873A (en) * | 2021-01-25 | 2021-06-04 | 中国农业科学院农产品加工研究所 | Free-state and conjugated-state salbutamol standard substance contained in swine urine freeze-dried powder after animal metabolism and preparation method thereof |
| CN112903875B (en) * | 2021-01-25 | 2023-02-17 | 中国农业科学院农产品加工研究所 | Ractopamine standard substance with conjugated state and free state coexisting in pig urine matrix as well as preparation method and application thereof |
| CN112903873B (en) * | 2021-01-25 | 2023-02-17 | 中国农业科学院农产品加工研究所 | Free-state and conjugated-state salbutamol standard substance contained in swine urine freeze-dried powder after animal metabolism and preparation method thereof |
| CN112903874B (en) * | 2021-01-25 | 2023-02-17 | 中国农业科学院农产品加工研究所 | Free and conjugated clenbuterol standard substance contained in pig urine and preparation method thereof |
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